U.S. patent application number 15/161137 was filed with the patent office on 2016-11-03 for methods and compositions for treating non-erk mapk pathway inhibitor-resistant cancers.
The applicant listed for this patent is BIOMED VALLEY DISCOVERIES, INC.. Invention is credited to Gary DeCrescenzo, Jeffrey James Roix, Saurabh Saha, Dean Welsch.
Application Number | 20160317519 15/161137 |
Document ID | / |
Family ID | 53403903 |
Filed Date | 2016-11-03 |
United States Patent
Application |
20160317519 |
Kind Code |
A1 |
Saha; Saurabh ; et
al. |
November 3, 2016 |
METHODS AND COMPOSITIONS FOR TREATING NON-ERK MAPK PATHWAY
INHIBITOR-RESISTANT CANCERS
Abstract
The present invention provides, inter alia, methods,
pharmaceutical compositions, and kits for treating or ameliorating
the effects of a cancer in a subject, which cancer is refractory or
resistant to non-ERK MAPK pathway inhibitor therapy. Also provided
are methods for identifying a subject having cancer who would
benefit from therapy with an ERK inhibitor and methods for
inhibiting phosphorylation of RSK in a cancer cell that is
refractory or resistant to a non-ERK MAPK pathway inhibitor.
Inventors: |
Saha; Saurabh; (Wellesley
Hills, MA) ; Welsch; Dean; (Parkville, MO) ;
DeCrescenzo; Gary; (Parkville, MO) ; Roix; Jeffrey
James; (Boston, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
BIOMED VALLEY DISCOVERIES, INC. |
KANSAS CITY |
MO |
US |
|
|
Family ID: |
53403903 |
Appl. No.: |
15/161137 |
Filed: |
May 20, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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PCT/US14/71749 |
Dec 19, 2014 |
|
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|
15161137 |
|
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|
61919551 |
Dec 20, 2013 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 31/12 20180101;
A61P 37/06 20180101; A61P 9/10 20180101; A61K 31/506 20130101; A61K
31/519 20130101; A61P 35/04 20180101; A61K 31/4439 20130101; G01N
33/57484 20130101; A61K 45/06 20130101; C12Q 2600/156 20130101;
A61P 43/00 20180101; C12Q 2600/106 20130101; A61P 35/00 20180101;
A61P 37/04 20180101; C12Q 1/6886 20130101; A61K 31/4439 20130101;
A61K 2300/00 20130101; A61K 31/506 20130101; A61K 2300/00 20130101;
A61K 31/519 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 31/4439 20060101
A61K031/4439; C12Q 1/68 20060101 C12Q001/68; A61K 31/519 20060101
A61K031/519; A61K 45/06 20060101 A61K045/06; A61K 31/506 20060101
A61K031/506 |
Claims
1. A method for treating or ameliorating the effects of a cancer in
a subject, which cancer is refractory or resistant to non-ERK MAPK
pathway inhibitor therapy, the method comprising administering to
the subject an effective amount of BVD-523 or a pharmaceutically
acceptable salt thereof.
2. The method according to claim 1, wherein the non-ERK MAPK
pathway inhibitor therapy is selected from the group consisting of
a RAS inhibitor, a RAF inhibitor, a MEK inhibitor, and combinations
thereof.
3. The method according to claim 1, wherein the non-ERK MAPK
pathway inhibitor therapy is selected from the group consisting of
a BRAF inhibitor, a MEK inhibitor, and combinations thereof.
4. The method according to claim 1, wherein substantially all
phosphorylation of RSK is inhibited after administration of BVD-523
or a pharmaceutically acceptable salt thereof.
5. The method according to claim 1, wherein the subject is a
mammal.
6. The method according to claim 5, wherein the mammal is selected
from the group consisting of humans, primates, farm animals, and
domestic animals.
7. The method according to claim 5, wherein the mammal is a
human.
8. The method according to claim 1, wherein the cancer has MAPK
activity.
9. The method according to claim 8, wherein the cancer is a solid
tumor cancer or a hematologic cancer.
10. The method according to claim 8, wherein the cancer is selected
from the group consisting of a cancer of the large intestine,
breast cancer, pancreatic cancer, skin cancer, and endometrial
cancers.
11. The method according to claim 8, wherein the cancer is
melanoma.
12. The method according to claim 1 further comprising
administering to the subject at least one additional therapeutic
agent selected from the group consisting of an antibody or fragment
thereof, a cytotoxic agent, a toxin, a radionuclide, an
immunomodulator, a photoactive therapeutic agent, a
radiosensitizing agent, a hormone, an anti-angiogenesis agent, and
combinations thereof.
13. The method according to claim 12, wherein the additional
therapeutic agent is an inhibitor of the PI3K/Akt pathway.
14. The method according to claim 13, wherein the inhibitor of the
PI3K/Akt pathway is selected from the group consisting of A-674563
(CAS #552325-73-2), AGL 2263, AMG-319 (Amgen, Thousand Oaks,
Calif.), AS-041164
(5-benzo[1,3]dioxol-5-ylmethylene-thiazolidine-2,4-dione),
AS-604850
(5-(2,2-Difluoro-benzo[1,3]dioxol-5-ylmethylene)-thiazolidine-2-
,4-dione), AS-605240
(5-quinoxilin-6-methylene-1,3-thiazolidine-2,4-dione), AT7867 (CAS
#857531-00-1), benzimidazole series, Genentech (Roche Holdings
Inc., South San Francisco, Calif.), BML-257 (CAS #32387-96-5),
CAL-120 (Gilead Sciences, Foster City, Calif.), CAL-129 (Gilead
Sciences), CAL-130 (Gilead Sciences), CAL-253 (Gilead Sciences),
CAL-263 (Gilead Sciences), CAS #612847-09-3, CAS #681281-88-9, CAS
#75747-14-7, CAS #925681-41-0, CAS #98510-80-6, CCT128930 (CAS
#885499-61-6), CH5132799 (CAS #1007207-67-1), CHR-4432 (Chroma
Therapeutics, Ltd., Abingdon, UK), FPA 124 (CAS #902779-59-3),
GS-1101 (CAL-101) (Gilead Sciences), GSK 690693 (CAS #937174-76-0),
H-89 (CAS #127243-85-0), Honokiol, IC87114 (Gilead Science),
IPI-145 (Intellikine Inc.), KAR-4139 (Karus Therapeutics,
Chilworth, UK), KAR-4141 (Karus Therapeutics), KIN-1 (Karus
Therapeutics), KT 5720 (CAS #108068-98-0), Miltefosine, MK-2206
dihydrochloride (CAS #1032350-13-2), ML-9 (CAS #105637-50-1),
Naltrindole Hydrochloride, OXY-111A (NormOxys Inc., Brighton,
Mass.), perifosine, PHT-427 (CAS #1191951-57-1), PI3 kinase delta
inhibitor, Merck KGaA (Merck & Co., Whitehouse Station, N.J.),
PI3 kinase delta inhibitors, Genentech (Roche Holdings Inc.), PI3
kinase delta inhibitors, Incozen (Incozen Therapeutics, Pvt. Ltd.,
Hydrabad, India), PI3 kinase delta inhibitors-2, Incozen (Incozen
Therapeutics), PI3 kinase inhibitor, Roche-4 (Roche Holdings Inc.),
PI3 kinase inhibitors, Roche (Roche Holdings Inc.), PI3 kinase
inhibitors, Roche-5 (Roche Holdings Inc.), PI3-alpha/delta
inhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd., South
San Francisco, Calif.), PI3-delta inhibitors, Cellzome (Cellzome
AG, Heidelberg, Germany), PI3-delta inhibitors, Intellikine
(Intellikine Inc., La Jolla, Calif.), PI3-delta inhibitors, Pathway
Therapeutics-1 (Pathway Therapeutics Ltd.), PI3-delta inhibitors,
Pathway Therapeutics-2 (Pathway Therapeutics Ltd.), PI3-delta/gamma
inhibitors, Cellzome (Cellzome AG), PI3-delta/gamma inhibitors,
Cellzome (Cellzome AG), PI3-delta/gamma inhibitors, Intellikine
(Intellikine Inc.), PI3-delta/gamma inhibitors, Intellikine
(Intellikine Inc.), PI3-delta/gamma inhibitors, Pathway
Therapeutics (Pathway Therapeutics Ltd.), PI3-delta/gamma
inhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd.),
PI3-gamma inhibitor Evotec (Evotec), PI3-gamma inhibitor, Cellzome
(Cellzome AG), PI3-gamma inhibitors, Pathway Therapeutics (Pathway
Therapeutics Ltd.), PI3K delta/gamma inhibitors, Intellikine-1
(Intellikine Inc.), PI3K delta/gamma inhibitors, Intellikine-1
(Intellikine Inc.), pictilisib (Roche Holdings Inc.), PIK-90 (CAS
#677338-12-4), SC-103980 (Pfizer, New York, N.Y.), SF-1126
(Semafore Pharmaceuticals, Indianapolis, Ind.), SH-5, SH-6,
Tetrahydro Curcumin, TG100-115 (Targegen Inc., San Diego, Calif.),
Triciribine, X-339 (Xcovery, West Palm Beach, Fla.), XL-499
(Evotech, Hamburg, Germany), pharmaceutically acceptable salts
thereof, and combinations thereof.
15. A method for treating or ameliorating the effects of a cancer
in a subject comprising: (a) identifying a subject with cancer that
has become refractory or resistant to BRAF inhibitor therapy, MEK
inhibitor therapy, or BRAF and MEK inhibitor therapy; and (b)
administering to the subject with said refractory or resistant
cancer an effective amount of an ERK inhibitor, which is BVD-523 or
a pharmaceutically acceptable salt thereof.
16. The method according to claim 15, wherein the subject is a
mammal.
17. The method according to claim 16, wherein the mammal is
selected from the group consisting of humans, primates, farm
animals, and domestic animals.
18. The method according to claim 16, wherein the mammal is a
human.
19. The method according to claim 15, wherein the cancer has MAPK
activity.
20. The method according to claim 19, wherein the cancer is a solid
tumor cancer or a hematologic cancer.
21. The method according to claim 19, wherein the cancer is
selected from the group consisting of a cancer of the large
intestine, breast cancer, pancreatic cancer, skin cancer, and
endometrial cancers.
22. The method according to claim 19, wherein the cancer is
melanoma.
23. The method according to claim 15, wherein identifying a subject
with cancer that is refractory or resistant to BRAF and/or MEK
inhibitor therapy comprises: (a) obtaining a biological sample from
the subject; and (b) screening the sample to determine whether the
subject has become resistant to an inhibitor therapy selected from
the group consisting of BRAF inhibitor therapy, MEK inhibitor
therapy, and combinations thereof.
24. The method according to claim 23, wherein the screening for a
cancer that is refractory or resistant to BRAF inhibitor therapy
comprises identifying (i) a switch between RAF isoforms, (ii)
upregulation of RTK or NRAS signaling, (iii) reactivation of
mitogen activated protein kinase (MAPK) signaling, (iv) the
presence of a MEK activating mutation, and combinations
thereof.
25. The method according to claim 23, wherein the screening for a
cancer that is refractory or resistant to MEK inhibitor therapy
comprises identifying (i) amplification of mutant BRAF, (ii) STAT3
upregulation, (iii) mutations in the allosteric pocket of MEK that
directly block binding of inhibitors to MEK or lead to constitutive
MEK activity, and combinations thereof.
26. The method according to claim 15 further comprising
administering at least one additional therapeutic agent selected
from the group consisting of an antibody or fragment thereof, a
cytotoxic agent, a toxin, a radionuclide, an immunomodulator, a
photoactive therapeutic agent, a radiosensitizing agent, a hormone,
an anti-angiogenesis agent, and combinations thereof.
27. The method according to claim 26, wherein the additional
therapeutic agent is an inhibitor of the PI3K/Akt pathway.
28. The method according to claim 27, wherein the inhibitor of the
PI3K/Akt pathway is selected from the group consisting of A-674563
(CAS #552325-73-2), AGL 2263, AMG-319 (Amgen, Thousand Oaks,
Calif.), AS-041164
(5-benzo[1,3]dioxol-5-ylmethylene-thiazolidine-2,4-dione),
AS-604850
(5-(2,2-Difluoro-benzo[1,3]dioxol-5-ylmethylene)-thiazolidine-2-
,4-dione), AS-605240
(5-quinoxilin-6-methylene-1,3-thiazolidine-2,4-dione), AT7867 (CAS
#857531-00-1), benzimidazole series, Genentech (Roche Holdings
Inc., South San Francisco, Calif.), BML-257 (CAS #32387-96-5),
CAL-120 (Gilead Sciences, Foster City, Calif.), CAL-129 (Gilead
Sciences), CAL-130 (Gilead Sciences), CAL-253 (Gilead Sciences),
CAL-263 (Gilead Sciences), CAS #612847-09-3, CAS #681281-88-9, CAS
#75747-14-7, CAS #925681-41-0, CAS #98510-80-6, CCT128930 (CAS
#885499-61-6), CH5132799 (CAS #1007207-67-1), CHR-4432 (Chroma
Therapeutics, Ltd., Abingdon, UK), FPA 124 (CAS #902779-59-3),
GS-1101 (CAL-101) (Gilead Sciences), GSK 690693 (CAS #937174-76-0),
H-89 (CAS #127243-85-0), Honokiol, IC87114 (Gilead Science),
IPI-145 (Intellikine Inc.), KAR-4139 (Karus Therapeutics,
Chilworth, UK), KAR-4141 (Karus Therapeutics), KIN-1 (Karus
Therapeutics), KT 5720 (CAS #108068-98-0), Miltefosine, MK-2206
dihydrochloride (CAS #1032350-13-2), ML-9 (CAS #105637-50-1),
Naltrindole Hydrochloride, OXY-111A (NormOxys Inc., Brighton,
Mass.), perifosine, PHT-427 (CAS #1191951-57-1), PI3 kinase delta
inhibitor, Merck KGaA (Merck & Co., Whitehouse Station, N.J.),
PI3 kinase delta inhibitors, Genentech (Roche Holdings Inc.), PI3
kinase delta inhibitors, Incozen (Incozen Therapeutics, Pvt. Ltd.,
Hydrabad, India), PI3 kinase delta inhibitors-2, Incozen (Incozen
Therapeutics), PI3 kinase inhibitor, Roche-4 (Roche Holdings Inc.),
PI3 kinase inhibitors, Roche (Roche Holdings Inc.), PI3 kinase
inhibitors, Roche-5 (Roche Holdings Inc.), PI3-alpha/delta
inhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd., South
San Francisco, Calif.), PI3-delta inhibitors, Cellzome (Cellzome
AG, Heidelberg, Germany), PI3-delta inhibitors, Intellikine
(Intellikine Inc., La Jolla, Calif.), PI3-delta inhibitors, Pathway
Therapeutics-1 (Pathway Therapeutics Ltd.), PI3-delta inhibitors,
Pathway Therapeutics-2 (Pathway Therapeutics Ltd.), PI3-delta/gamma
inhibitors, Cellzome (Cellzome AG), PI3-delta/gamma inhibitors,
Cellzome (Cellzome AG), PI3-delta/gamma inhibitors, Intellikine
(Intellikine Inc.), PI3-delta/gamma inhibitors, Intellikine
(Intellikine Inc.), PI3-delta/gamma inhibitors, Pathway
Therapeutics (Pathway Therapeutics Ltd.), PI3-delta/gamma
inhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd.),
PI3-gamma inhibitor Evotec (Evotec), PI3-gamma inhibitor, Cellzome
(Cellzome AG), PI3-gamma inhibitors, Pathway Therapeutics (Pathway
Therapeutics Ltd.), PI3K delta/gamma inhibitors, Intellikine-1
(Intellikine Inc.), PI3K delta/gamma inhibitors, Intellikine-1
(Intellikine Inc.), pictilisib (Roche Holdings Inc.), PIK-90 (CAS
#677338-12-4), SC-103980 (Pfizer, New York, N.Y.), SF-1126
(Semafore Pharmaceuticals, Indianapolis, Ind.), SH-5, SH-6,
Tetrahydro Curcumin, TG100-115 (Targegen Inc., San Diego, Calif.),
Triciribine, X-339 (Xcovery, West Palm Beach, Fla.), XL-499
(Evotech, Hamburg, Germany), pharmaceutically acceptable salts
thereof, and combinations thereof.
29. A method for treating or ameliorating the effects of cancer in
a subject, which cancer is refractory or resistant to BRAF
inhibitor therapy, MEK inhibitor therapy, or both, the method
comprising administering to the subject an effective amount of
BVD-523 or a pharmaceutically acceptable salt thereof.
30. The method according to claim 29, wherein the subject is a
mammal.
31. The method according to claim 30, wherein the mammal is
selected from the group consisting of humans, primates, farm
animals, and domestic animals.
32. The method according to claim 30, wherein the mammal is a
human.
33. The method according to claim 29, wherein the cancer has MAPK
activity.
34. The method according to claim 33, wherein the cancer is a solid
tumor cancer or a hematologic cancer.
35. The method according to claim 33, wherein the cancer is
selected from the group consisting of a cancer of the large
intestine, breast cancer, pancreatic cancer, skin cancer, and
endometrial cancers.
36. The method according to claim 33, wherein the cancer is
melanoma.
37. The method according to claim 29, wherein the cancer is
determined to be refractory or resistant to BRAF inhibitor therapy
based on one or more of the following: (i) a switch between RAF
isoforms, (ii) upregulation of RTK or NRAS signaling, (iii)
reactivation of mitogen activated protein kinase (MAPK) signaling,
(iv) the presence of a MEK activating mutation.
38. The method according to claim 29, wherein the cancer is
determined to be refractory or resistant to MEK inhibitor therapy
based on one or more of the following: (i) amplification of mutant
BRAF, (ii) STAT3 upregulation, (iii) mutations in the allosteric
pocket of MEK that directly block binding of inhibitors to MEK or
lead to constitutive MEK activity.
39. The method according to claim 29 further comprising
administering to the subject at least one additional therapeutic
agent selected from the group consisting of an antibody or fragment
thereof, a cytotoxic agent, a toxin, a radionuclide, an
immunomodulator, a photoactive therapeutic agent, a
radiosensitizing agent, a hormone, an anti-angiogenesis agent, and
combinations thereof.
40. The method according to claim 39, wherein the additional
therapeutic agent is an inhibitor of the PI3K/Akt pathway.
41. The method according to claim 40, wherein the inhibitor of the
PI3K/Akt pathway is selected from the group consisting of A-674563
(CAS #552325-73-2), AGL 2263, AMG-319 (Amgen, Thousand Oaks,
Calif.), AS-041164
(5-benzo[1,3]dioxol-5-ylmethylene-thiazolidine-2,4-dione),
AS-604850
(5-(2,2-Difluoro-benzo[1,3]dioxol-5-ylmethylene)-thiazolidine-2-
,4-dione), AS-605240
(5-quinoxilin-6-methylene-1,3-thiazolidine-2,4-dione), AT7867 (CAS
#857531-00-1), benzimidazole series, Genentech (Roche Holdings
Inc., South San Francisco, Calif.), BML-257 (CAS #32387-96-5),
CAL-120 (Gilead Sciences, Foster City, Calif.), CAL-129 (Gilead
Sciences), CAL-130 (Gilead Sciences), CAL-253 (Gilead Sciences),
CAL-263 (Gilead Sciences), CAS #612847-09-3, CAS #681281-88-9, CAS
#75747-14-7, CAS #925681-41-0, CAS #98510-80-6, CCT128930 (CAS
#885499-61-6), CH5132799 (CAS #1007207-67-1), CHR-4432 (Chroma
Therapeutics, Ltd., Abingdon, UK), FPA 124 (CAS #902779-59-3),
GS-1101 (CAL-101) (Gilead Sciences), GSK 690693 (CAS #937174-76-0),
H-89 (CAS #127243-85-0), Honokiol, IC87114 (Gilead Science),
IPI-145 (Intellikine Inc.), KAR-4139 (Karus Therapeutics,
Chilworth, UK), KAR-4141 (Karus Therapeutics), KIN-1 (Karus
Therapeutics), KT 5720 (CAS #108068-98-0), Miltefosine, MK-2206
dihydrochloride (CAS #1032350-13-2), ML-9 (CAS #105637-50-1),
Naltrindole Hydrochloride, OXY-111A (NormOxys Inc., Brighton,
Mass.), perifosine, PHT-427 (CAS #1191951-57-1), PI3 kinase delta
inhibitor, Merck KGaA (Merck & Co., Whitehouse Station, N.J.),
PI3 kinase delta inhibitors, Genentech (Roche Holdings Inc.), PI3
kinase delta inhibitors, Incozen (Incozen Therapeutics, Pvt. Ltd.,
Hydrabad, India), PI3 kinase delta inhibitors-2, Incozen (Incozen
Therapeutics), PI3 kinase inhibitor, Roche-4 (Roche Holdings Inc.),
PI3 kinase inhibitors, Roche (Roche Holdings Inc.), PI3 kinase
inhibitors, Roche-5 (Roche Holdings Inc.), PI3-alpha/delta
inhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd., South
San Francisco, Calif.), PI3-delta inhibitors, Cellzome (Cellzome
AG, Heidelberg, Germany), PI3-delta inhibitors, Intellikine
(Intellikine Inc., La Jolla, Calif.), PI3-delta inhibitors, Pathway
Therapeutics-1 (Pathway Therapeutics Ltd.), PI3-delta inhibitors,
Pathway Therapeutics-2 (Pathway Therapeutics Ltd.), PI3-delta/gamma
inhibitors, Cellzome (Cellzome AG), PI3-delta/gamma inhibitors,
Cellzome (Cellzome AG), PI3-delta/gamma inhibitors, Intellikine
(Intellikine Inc.), PI3-delta/gamma inhibitors, Intellikine
(Intellikine Inc.), PI3-delta/gamma inhibitors, Pathway
Therapeutics (Pathway Therapeutics Ltd.), PI3-delta/gamma
inhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd.),
PI3-gamma inhibitor Evotec (Evotec), PI3-gamma inhibitor, Cellzome
(Cellzome AG), PI3-gamma inhibitors, Pathway Therapeutics (Pathway
Therapeutics Ltd.), PI3K delta/gamma inhibitors, Intellikine-1
(Intellikine Inc.), PI3K delta/gamma inhibitors, Intellikine-1
(Intellikine Inc.), pictilisib (Roche Holdings Inc.), PIK-90 (CAS
#677338-12-4), SC-103980 (Pfizer, New York, N.Y.), SF-1126
(Semafore Pharmaceuticals, Indianapolis, Ind.), SH-5, SH-6,
Tetrahydro Curcumin, TG100-115 (Targegen Inc., San Diego, Calif.),
Triciribine, X-339 (Xcovery, West Palm Beach, Fla.), XL-499
(Evotech, Hamburg, Germany), pharmaceutically acceptable salts
thereof, and combinations thereof.
42. A method for identifying a subject having cancer who would
benefit from therapy with an ERK inhibitor, the method comprising:
(a) obtaining a biological sample from the subject; and (b)
screening the sample to determine whether the subject has one or
more of the following markers: (i) a switch between RAF isoforms,
(ii) upregulation of receptor tyrosine kinase (RTK) or NRAS
signaling, (iii) reactivation of mitogen activated protein kinase
(MAPK) signaling, (iv) the presence of a MEK activating mutation,
(v) amplification of mutant BRAF, (vi) STAT3 upregulation, (vii)
mutations in the allosteric pocket of MEK that directly block
binding of inhibitors to MEK or lead to constitutive MEK activity,
wherein the presence of one or more of the markers confirms that
the subject's cancer is refractory or resistant to BRAF and/or MEK
inhibitor therapy and that the subject would benefit from therapy
with an ERK inhibitor, which is BVD-523 or a pharmaceutically
acceptable salt thereof.
43. The method according to claim 42, wherein the subject is a
mammal.
44. The method according to claim 43, wherein the mammal is
selected from the group consisting of humans, primates, farm
animals, and domestic animals.
45. The method according to claim 43, wherein the mammal is a
human.
46. The method according to claim 42, wherein the cancer has MAPK
activity.
47. The method according to claim 46, wherein the cancer is a solid
tumor cancer or a hematologic cancer.
48. The method according to claim 46, wherein the cancer is
selected from the group consisting of a cancer of the large
intestine, breast cancer, pancreatic cancer, skin cancer, and
endometrial cancers.
49. The method according to claim 46, wherein the cancer is
melanoma.
50. The method according to claim 42 further comprising
administering BVD-523 or a pharmaceutically acceptable salt thereof
to a subject having one or more of the markers.
51. The method according to claim 50 further comprising
administering to the subject having one or more of the markers at
least one additional therapeutic agent selected from the group
consisting of an antibody or fragment thereof, a cytotoxic agent, a
toxin, a radionuclide, an immunomodulator, a photoactive
therapeutic agent, a radiosensitizing agent, a hormone, an
anti-angiogenesis agent, and combinations thereof.
52. The method according to claim 51, wherein the additional
therapeutic agent is an inhibitor of the PI3K/Akt pathway.
53. The method according to claim 52, wherein the inhibitor of the
PI3K/Akt pathway is selected from the group consisting of A-674563
(CAS #552325-73-2), AGL 2263, AMG-319 (Amgen, Thousand Oaks,
Calif.), AS-041164
(5-benzo[1,3]dioxol-5-ylmethylene-thiazolidine-2,4-dione),
AS-604850
(5-(2,2-Difluoro-benzo[1,3]dioxol-5-ylmethylene)-thiazolidine-2-
,4-dione), AS-605240
(5-quinoxilin-6-methylene-1,3-thiazolidine-2,4-dione), AT7867 (CAS
#857531-00-1), benzimidazole series, Genentech (Roche Holdings
Inc., South San Francisco, Calif.), BML-257 (CAS #32387-96-5),
CAL-120 (Gilead Sciences, Foster City, Calif.), CAL-129 (Gilead
Sciences), CAL-130 (Gilead Sciences), CAL-253 (Gilead Sciences),
CAL-263 (Gilead Sciences), CAS #612847-09-3, CAS #681281-88-9, CAS
#75747-14-7, CAS #925681-41-0, CAS #98510-80-6, CCT128930 (CAS
#885499-61-6), CH5132799 (CAS #1007207-67-1), CHR-4432 (Chroma
Therapeutics, Ltd., Abingdon, UK), FPA 124 (CAS #902779-59-3),
GS-1101 (CAL-101) (Gilead Sciences), GSK 690693 (CAS #937174-76-0),
H-89 (CAS #127243-85-0), Honokiol, IC87114 (Gilead Science),
IPI-145 (Intellikine Inc.), KAR-4139 (Karus Therapeutics,
Chilworth, UK), KAR-4141 (Karus Therapeutics), KIN-1 (Karus
Therapeutics), KT 5720 (CAS #108068-98-0), Miltefosine, MK-2206
dihydrochloride (CAS #1032350-13-2), ML-9 (CAS #105637-50-1),
Naltrindole Hydrochloride, OXY-111A (NormOxys Inc., Brighton,
Mass.), perifosine, PHT-427 (CAS #1191951-57-1), PI3 kinase delta
inhibitor, Merck KGaA (Merck & Co., Whitehouse Station, N.J.),
PI3 kinase delta inhibitors, Genentech (Roche Holdings Inc.), PI3
kinase delta inhibitors, Incozen (Incozen Therapeutics, Pvt. Ltd.,
Hydrabad, India), PI3 kinase delta inhibitors-2, Incozen (Incozen
Therapeutics), PI3 kinase inhibitor, Roche-4 (Roche Holdings Inc.),
PI3 kinase inhibitors, Roche (Roche Holdings Inc.), PI3 kinase
inhibitors, Roche-5 (Roche Holdings Inc.), PI3-alpha/delta
inhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd., South
San Francisco, Calif.), PI3-delta inhibitors, Cellzome (Cellzome
AG, Heidelberg, Germany), PI3-delta inhibitors, Intellikine
(Intellikine Inc., La Jolla, Calif.), PI3-delta inhibitors, Pathway
Therapeutics-1 (Pathway Therapeutics Ltd.), PI3-delta inhibitors,
Pathway Therapeutics-2 (Pathway Therapeutics Ltd.), PI3-delta/gamma
inhibitors, Cellzome (Cellzome AG), PI3-delta/gamma inhibitors,
Cellzome (Cellzome AG), PI3-delta/gamma inhibitors, Intellikine
(Intellikine Inc.), PI3-delta/gamma inhibitors, Intellikine
(Intellikine Inc.), PI3-delta/gamma inhibitors, Pathway
Therapeutics (Pathway Therapeutics Ltd.), PI3-delta/gamma
inhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd.),
PI3-gamma inhibitor Evotec (Evotec), PI3-gamma inhibitor, Cellzome
(Cellzome AG), PI3-gamma inhibitors, Pathway Therapeutics (Pathway
Therapeutics Ltd.), PI3K delta/gamma inhibitors, Intellikine-1
(Intellikine Inc.), PI3K delta/gamma inhibitors, Intellikine-1
(Intellikine Inc.), pictilisib (Roche Holdings Inc.), PIK-90 (CAS
#677338-12-4), SC-103980 (Pfizer, New York, N.Y.), SF-1126
(Semafore Pharmaceuticals, Indianapolis, Ind.), SH-5, SH-6,
Tetrahydro Curcumin, TG100-115 (Targegen Inc., San Diego, Calif.),
Triciribine, X-339 (Xcovery, West Palm Beach, Fla.), XL-499
(Evotech, Hamburg, Germany), pharmaceutically acceptable salts
thereof, and combinations thereof.
54. A pharmaceutical composition for treating or ameliorating the
effects of a cancer in a subject, which cancer is refractory or
resistant to non-ERK MAPK pathway therapy, the composition
comprising a pharmaceutically acceptable carrier or diluent and an
effective amount of BVD-523 or a pharmaceutically acceptable salt
thereof.
55. The pharmaceutical composition according to claim 54, wherein
the non-ERK MAPK pathway inhibitor therapy is selected from the
group consisting of a RAS inhibitor, a RAF inhibitor, a MEK
inhibitor, and combinations thereof.
56. The pharmaceutical composition according to claim 54, wherein
the non-ERK MAPK pathway inhibitor therapy is selected from the
group consisting of a BRAF inhibitor, a MEK inhibitor, and
combinations thereof.
57. The pharmaceutical composition according to claim 54, wherein
the subject is a mammal.
58. The pharmaceutical composition according to claim 57, wherein
the mammal is selected from the group consisting of humans,
primates, farm animals, and domestic animals.
59. The pharmaceutical composition according to claim 57, wherein
the mammal is a human.
60. The pharmaceutical composition according to claim 54, wherein
the cancer has MAPK activity.
61. The pharmaceutical composition according to claim 60, wherein
the cancer is a solid tumor cancer or a hematologic cancer.
62. The pharmaceutical composition according to claim 60, wherein
the cancer is selected from the group consisting of a cancer of the
large intestine, breast cancer, pancreatic cancer, skin cancer, and
endometrial cancers.
63. The pharmaceutical composition according to claim 60, wherein
the cancer is melanoma.
64. The pharmaceutical composition according to claim 54 further
comprising at least one additional therapeutic agent selected from
the group consisting of an antibody or fragment thereof, a
cytotoxic agent, a toxin, a radionuclide, an immunomodulator, a
photoactive therapeutic agent, a radiosensitizing agent, a hormone,
an anti-angiogenesis agent, and combinations thereof.
65. The pharmaceutical composition according to claim 64, wherein
the additional therapeutic agent is an inhibitor of the PI3K/Akt
pathway.
66. The pharmaceutical composition according to claim 65, wherein
the inhibitor of the PI3K/Akt pathway is selected from the group
consisting of A-674563 (CAS #552325-73-2), AGL 2263, AMG-319
(Amgen, Thousand Oaks, Calif.), AS-041164
(5-benzo[1,3]dioxol-5-ylmethylene-thiazolidine-2,4-dione),
AS-604850
(5-(2,2-Difluoro-benzo[1,3]dioxol-5-ylmethylene)-thiazolidine-2,4-dione),
AS-605240 (5-quinoxilin-6-methylene-1,3-thiazolidine-2,4-dione),
AT7867 (CAS #857531-00-1), benzimidazole series, Genentech (Roche
Holdings Inc., South San Francisco, Calif.), BML-257 (CAS
#32387-96-5), CAL-120 (Gilead Sciences, Foster City, Calif.),
CAL-129 (Gilead Sciences), CAL-130 (Gilead Sciences), CAL-253
(Gilead Sciences), CAL-263 (Gilead Sciences), CAS #612847-09-3, CAS
#681281-88-9, CAS #75747-14-7, CAS #925681-41-0, CAS #98510-80-6,
CCT128930 (CAS #885499-61-6), CH5132799 (CAS #1007207-67-1),
CHR-4432 (Chroma Therapeutics, Ltd., Abingdon, UK), FPA 124 (CAS
#902779-59-3), GS-1101 (CAL-101) (Gilead Sciences), GSK 690693 (CAS
#937174-76-0), H-89 (CAS #127243-85-0), Honokiol, IC87114 (Gilead
Science), IPI-145 (Intellikine Inc.), KAR-4139 (Karus Therapeutics,
Chilworth, UK), KAR-4141 (Karus Therapeutics), KIN-1 (Karus
Therapeutics), KT 5720 (CAS #108068-98-0), Miltefosine, MK-2206
dihydrochloride (CAS #1032350-13-2), ML-9 (CAS #105637-50-1),
Naltrindole Hydrochloride, OXY-111A (NormOxys Inc., Brighton,
Mass.), perifosine, PHT-427 (CAS #1191951-57-1), PI3 kinase delta
inhibitor, Merck KGaA (Merck & Co., Whitehouse Station, N.J.),
PI3 kinase delta inhibitors, Genentech (Roche Holdings Inc.), PI3
kinase delta inhibitors, Incozen (Incozen Therapeutics, Pvt. Ltd.,
Hydrabad, India), PI3 kinase delta inhibitors-2, Incozen (Incozen
Therapeutics), PI3 kinase inhibitor, Roche-4 (Roche Holdings Inc.),
PI3 kinase inhibitors, Roche (Roche Holdings Inc.), PI3 kinase
inhibitors, Roche-5 (Roche Holdings Inc.), PI3-alpha/delta
inhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd., South
San Francisco, Calif.), PI3-delta inhibitors, Cellzome (Cellzome
AG, Heidelberg, Germany), PI3-delta inhibitors, Intellikine
(Intellikine Inc., La Jolla, Calif.), PI3-delta inhibitors, Pathway
Therapeutics-1 (Pathway Therapeutics Ltd.), PI3-delta inhibitors,
Pathway Therapeutics-2 (Pathway Therapeutics Ltd.), PI3-delta/gamma
inhibitors, Cellzome (Cellzome AG), PI3-delta/gamma inhibitors,
Cellzome (Cellzome AG), PI3-delta/gamma inhibitors, Intellikine
(Intellikine Inc.), PI3-delta/gamma inhibitors, Intellikine
(Intellikine Inc.), PI3-delta/gamma inhibitors, Pathway
Therapeutics (Pathway Therapeutics Ltd.), PI3-delta/gamma
inhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd.),
PI3-gamma inhibitor Evotec (Evotec), PI3-gamma inhibitor, Cellzome
(Cellzome AG), PI3-gamma inhibitors, Pathway Therapeutics (Pathway
Therapeutics Ltd.), PI3K delta/gamma inhibitors, Intellikine-1
(Intellikine Inc.), PI3K delta/gamma inhibitors, Intellikine-1
(Intellikine Inc.), pictilisib (Roche Holdings Inc.), PIK-90 (CAS
#677338-12-4), SC-103980 (Pfizer, New York, N.Y.), SF-1126
(Semafore Pharmaceuticals, Indianapolis, Ind.), SH-5, SH-6,
Tetrahydro Curcumin, TG100-115 (Targegen Inc., San Diego, Calif.),
Triciribine, X-339 (Xcovery, West Palm Beach, Fla.), XL-499
(Evotech, Hamburg, Germany), pharmaceutically acceptable salts
thereof, and combinations thereof.
67. A kit for treating or ameliorating the effects of a cancer in a
subject, which cancer is refractory or resistant to non-ERK MAPK
pathway therapy, the kit comprising a pharmaceutical composition
according to any one of claims 54, 55, or 56 packaged together with
instructions for its use.
68. A method for inhibiting phosphorylation of RSK in a cancer cell
that is refractory or resistant to a non-ERK MAPK pathway
inhibitor, the method comprising contacting the cancer cell with an
effective amount of BVD-523 or a pharmaceutically acceptable salt
thereof for a period of time sufficient for phosphorylation of RSK
in the cancer cell to be inhibited.
69. The method according to claim 68, wherein greater than 50% of
RSK phosphorylation is inhibited.
70. The method according to claim 68, wherein greater than 75% of
RSK phosphorylation is inhibited.
71. The method according to claim 68, wherein greater than 90% of
RSK phosphorylation is inhibited.
72. The method according to claim 68, wherein greater than 95% of
RSK phosphorylation is inhibited.
73. The method according to claim 68, wherein greater than 99% of
RSK phosphorylation is inhibited.
74. The method according to claim 68, wherein 100% of RSK
phosphorylation is inhibited.
75. The method according to claim 68, which is carried out in
vitro, ex vivo, or in vivo.
76. The method according to claim 68, wherein the contacting step
comprises administering BVD-523 or a pharmaceutically acceptable
salt to a subject from whom the cancer cell was obtained.
77. The method according to claim 68, wherein the non-ERK MAPK
pathway inhibitor is selected from the group consisting of a RAS
inhibitor, a RAF inhibitor, a MEK inhibitor, and combinations
thereof.
78. The method according to claim 68, wherein the non-ERK MAPK
pathway inhibitor is selected from the group consisting of BRAF
inhibitors, MEK inhibitors, and combinations thereof.
79. The method according to claim 68, wherein the cancer is from a
mammal.
80. The method according to claim 79, wherein the mammal is
selected from the group consisting of humans, primates, farm
animals, and domestic animals.
81. The method according to claim 79, wherein the mammal is a
human.
82. The method according to claim 68, wherein the cancer has MAPK
activity.
83. The method according to claim 82, wherein the cancer is a solid
tumor cancer or a hematologic cancer.
84. The method according to claim 82, wherein the cancer is
selected from the group consisting of a cancer of the large
intestine, breast cancer, pancreatic cancer, skin cancer, and
endometrial cancers.
85. The method according to claim 82, wherein the cancer is
melanoma.
86. A method of treating a subject having an unresectable or
metastatic BRAF600 mutation-positive melanoma comprising
administering to the subject 600 mg BID of BVD-523 or a
pharmaceutically acceptable salt thereof.
87. The method according to claim 86, wherein the mutation is a
BRAF.sup.V600E mutation.
88. The method according to claim 86, wherein the mammal is
selected from the group consisting of humans, primates, farm
animals, and domestic animals.
89. The method according to claim 86, wherein the mammal is a
human.
90. The method according to claim 86, wherein the melanoma has MAPK
activity.
91. A composition for treating a subject having an unresectable or
metastatic BRAF600 mutation-positive melanoma, the composition
comprising 600 mg of BVD-523 or a pharmaceutically acceptable salt
thereof and optionally a pharmaceutically acceptable carrier,
adjuvant, or vehicle.
92. The composition according to claim 91, wherein the subject is a
mammal.
93. The composition according to claim 91, wherein the mammal is
selected from the group consisting of humans, primates, farm
animals, and domestic animals.
94. The composition according to claim 91, wherein the mammal is a
human.
95. The composition according to claim 91, wherein the melanoma has
MAPK activity.
96. The composition of claim 91 wherein the mutation is a
BRAF.sup.V600E mutation.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation in part of PCT
international application no. PCT/US2014/071749, filed Dec. 19,
2014, which claims benefit of U.S. Patent Application Ser. No.
61/919,551, filed on Dec. 20, 2013 which, applications are
incorporated by reference herein in their entireties.
FIELD OF INVENTION
[0002] The present invention provides, inter alia, methods,
pharmaceutical compositions, and kits for treating or ameliorating
the effects of a cancer in a subject, which cancer is refractory or
resistant to non-ERK MAPK pathway inhibitor therapy.
INCORPORATION BY REFERENCE OF SEQUENCE LISTING
[0003] This application contains references to amino acids and/or
nucleic acid sequences that have been filed concurrently herewith
as sequence listing text file "0375608.txt", file size of 356 KB,
created on Dec. 18, 2014. The aforementioned sequence listing is
hereby incorporated by reference in its entirety pursuant to 37
C.F.R. .sctn.1.52(e)(5).
BACKGROUND OF THE INVENTION
[0004] Drug inhibitors that target components of the
mitogen-activated protein kinases (MAPK) signaling pathway show
clinical efficacy in a variety of cancers, particularly those
bearing mutations in the BRAF protein kinase. Both RAF and MEK
inhibitors are approved for single-agent use in advanced metastatic
BRAF mutant melanoma. Either alone or in combination, BRAF and MEK
inhibitor activity is unpredictable in other cancers, with
promising efficacy in BRAF mutant thyroid and lung cancer, but only
marginal activity in BRAF mutant colorectal cancer.
[0005] As with other targeted therapies, patterns of disease
response to RAF and MEK inhibitors appear to be influenced by the
intrinsic genetic heterogeneity present in the cancers where the
drugs are used. For instance, it has been shown that certain
genetic alterations, including PTEN and other changes that activate
the PI3K cell growth signaling pathway, may predict a poor initial
response, and/or relatively rapid progression, in BRAF mutant
melanoma treated with the RAF inhibitor vemurafenib. Likewise,
direct mutations in MEK gene loci appear to emerge in tumors that
have progressed following either BRAF, MEK, or combined drug
treatment. Several additional examples, from RAS and RAF gene
amplification and splicing mutations, suggest that acquired drug
resistance is produced when oncogenic pleiotropy encounters the
selective pressure of targeted drug treatment.
[0006] In view of the foregoing, there is a need for novel targeted
agents that would ideally inhibit diverse nodes of oncogenic
pathways, and also be effective in combinations by inducing a
burden of selective pressures that exceeds the adaptive capacity of
diverse cancer genomes. The present application is directed to
meeting these and other needs.
SUMMARY OF THE INVENTION
[0007] One embodiment of the present invention is a method for
treating or ameliorating the effects of a cancer in a subject,
which cancer is refractory or resistant to non-ERK MAPK pathway
inhibitor therapy. The method comprises administering to the
subject an effective amount of BVD-523 or a pharmaceutically
acceptable salt thereof.
[0008] Another embodiment of the present invention is a method for
treating or ameliorating the effects of a cancer in a subject. The
method comprises: [0009] (a) identifying a subject with cancer that
has become refractory or resistant to BRAF inhibitor therapy, MEK
inhibitor therapy, or BRAF and MEK inhibitor therapy; and [0010]
(b) administering to the subject with said refractory or resistant
cancer an effective amount of an ERK inhibitor, which is BVD-523 or
a pharmaceutically acceptable salt thereof.
[0011] A further embodiment of the present invention is a method
for treating or ameliorating the effects of cancer in a subject,
which cancer is refractory or resistant to BRAF inhibitor therapy,
MEK inhibitor therapy, or both. The method comprises administering
to the subject an effective amount of BVD-523 or a pharmaceutically
acceptable salt thereof.
[0012] Another embodiment of the present invention is a method for
identifying a subject having cancer who would benefit from therapy
with an ERK inhibitor. The method comprises: [0013] (a) obtaining a
biological sample from the subject; and [0014] (b) screening the
sample to determine whether the subject has one or more of the
following markers: [0015] (i) a switch between RAF isoforms, [0016]
(ii) upregulation of receptor tyrosine kinase (RTK) or NRAS
signaling, [0017] (iii) reactivation of mitogen activated protein
kinase (MAPK) signaling, [0018] (iv) the presence of a MEK
activating mutation, [0019] (v) amplification of mutant BRAF,
[0020] (vi) STAT3 upregulation, [0021] (vii) mutations in the
allosteric pocket of MEK that directly block binding of inhibitors
to MEK or lead to constitutive MEK activity, wherein the presence
of one or more of the markers confirms that the subject's cancer is
refractory or resistant to BRAF and/or MEK inhibitor therapy and
that the subject would benefit from therapy with an ERK inhibitor,
which is BVD-523 or a pharmaceutically acceptable salt thereof.
[0022] A further embodiment of the present invention is a
pharmaceutical composition for treating or ameliorating the effects
of a cancer in a subject, which cancer is refractory or resistant
to non-ERK MAPK pathway therapy. The composition comprises a
pharmaceutically acceptable carrier or diluent and an effective
amount of BVD-523 or a pharmaceutically acceptable salt
thereof.
[0023] Another embodiment of the present invention is a kit for
treating or ameliorating the effects of a cancer in a subject,
which cancer is refractory or resistant to non-ERK MAPK pathway
therapy. The kit comprises any of the pharmaceutical compositions
according to the present invention packaged together with
instructions for its use.
[0024] Another embodiment of the present invention is a method for
inhibiting phosphorylation of RSK in a cancer cell that is
refractory or resistant to a non-ERK MAPK pathway inhibitor. The
method comprises contacting the cancer cell with an effective
amount of BVD-523 or a pharmaceutically acceptable salt thereof for
a period of time sufficient for phosphorylation of RSK in the
cancer cell to be inhibited.
[0025] Another embodiment of the present invention is a method of
treating a subject having an unresectable or metastatic BRAF600
mutation-positive melanoma comprising administering to the subject
600 mg BID of BVD-523 or a pharmaceutically acceptable salt
thereof.
[0026] Another embodiment of the present invention is a composition
for treating a subject having an unresectable or metastatic BRAF600
mutation-positive melanoma, the composition comprising 600 mg of
BVD-523 or a pharmaceutically acceptable salt thereof and
optionally a pharmaceutically acceptable carrier, adjuvant, or
vehicle.
BRIEF DESCRIPTION OF THE DRAWINGS
[0027] The patent or application file contains at least one drawing
executed in color. Copies of this patent or patent application
publication with color drawing(s) will be provided by the Office
upon request and payment of the necessary fee.
[0028] FIG. 1A-FIG. 1C show the progress of a dose escalation study
in a human malignant melanoma cell line (A375 cells) for month 1.
Various treatments (trametinib (a type 2 MEK inhibitor), dabrafenib
(a BRAF inhibitor), and BVD-523 (an ERK1/2 inhibior)) are as
labeled.
[0029] FIG. 2A-FIG. 2H show the results of a proliferation assay
that tracks changes in sensitivity to the escalated agent(s) at
month 1. Various treatments (trametinib, dabrafenib, BVD-523, and
pacitaxel) are as labeled on the top of the graph. The caption to
the right of the graph shows the various types of cells generated
from the dose escalation study. For example, "dabrafenib" refers to
the cells that have been treated with the highest dose of
dabrafenib from month 1 of the dose escalation study. Parental
refers to the control cells that have not been treated with drugs.
FIG. 2A, FIG. 2C and FIG. 2G are normalized to control, whereas
FIG. 2D, FIG. 2F and FIG. 2H show the raw data.
[0030] FIG. 3A-FIG. 3D show the progress of a dose escalation study
in A375 cells for month 2. Various treatments (trametinib,
dabrafenib, and BVD-523) are as labeled.
[0031] FIG. 4A-FIG. 4H show the results of a proliferation assay
that tracks changes in sensitivity to the escalated agent(s) at
month 2. Various treatments (trametinib, dabrafenib, BVD-523, and
pacitaxel) are as labeled on the top of the graph. The caption to
the right of the graph shows the various types of cells generated
from the dose escalation study. For example, "dabrafenib" refers to
the cells that have been treated with the highest dose of
dabrafenib from month 2 of the dose escalation study. Parental
refers to the control cells that have not been treated with drugs.
FIG. 4A, FIG. 4C and FIG. 4G are normalized to control, whereas
FIG. 4D, FIG. 4F and FIG. 4H show the raw data.
[0032] FIG. 5A-FIG. 5H show only the parental and BVD-523 cell line
data from FIG. 4A-FIG. 4H. Various treatments (trametinib,
dabrafenib, BVD-523, and pacitaxel) are as labeled. FIG. 5A, FIG.
5C and FIG. 5G are normalized to control, whereas FIG. 5D, FIG. 5F
and FIG. 5H show the raw data.
[0033] FIG. 6A-FIG. 6D show the progress of the dose escalation
study in a human malignant cell line (A375 cells) for month 3.
Various treatments (trametinib, dabrafenib, and BVD-523) are as
labeled.
[0034] FIG. 7 is a histogram showing the results of a proliferation
assay as applied to cells grown in the DMSO control wells from the
dose escalation assay.
[0035] FIG. 8A-FIG. 8D are a set of line graphs showing
proliferation assays for month 3 of the study. Various treatments
(trametinib, dabrafenib, BVD-523, and pacitaxel) are as labeled on
the top of the graph. The caption to the right of the graph shows
the various types of cells generated from the dose escalation
study. For example, "dabrafenib" refers to the cells that have been
treated with the highest dose of dabrafenib from month 3 of the
dose escalation study. Parental refers to the control cells that
have not been treated with drugs.
[0036] FIG. 9A-FIG. 9D show only the parental, dabrafenib, and
BVD-523 cell line data from FIG. 8A-FIG. 8D.
[0037] FIG. 10A is a dose matrix showing % inhibition of the
trametinib/dabrafenib combination in A375 cells using the Alamar
Blue cell viability assay. FIG. 10B is a dose matrix showing excess
over Bliss for the trametinib/dabrafenib combination. FIG. 10C and
FIG. 10D show % viability relative to DMSO only treated controls
for dabrafenib and trametinib single agent treatments in A375 cells
using the Alamar Blue cell viability assay. FIG. 10E shows %
viability relative to DMSO only treated controls for dabrafenib and
trametinib combination treatments in A375 cells using the Alamar
Blue cell viability assay.
[0038] FIG. 11A is a dose matrix showing % inhibition of the
trametinib/dabrafenib combination in A375 cells using the
CellTiter-Glo cell viability assay. FIG. 11B is a dose matrix
showing excess over Bliss for the trametinib/dabrafenib
combination. FIG. 11C and FIG. 11D show % viability relative to
DMSO only treated controls for dabrafenib and trametinib single
agent treatments in A375 cells using the CellTiter-Glo cell
viability assay. FIG. 11E shows % viability relative to DMSO only
treated controls for dabrafenib and trametinib combination
treatments in A375 cells using the CellTiter-Glo cell viability
assay.
[0039] FIG. 12A is a dose matrix showing % inhibition of the
BVD-523/dabrafenib combination in A375 cells using the Alamar Blue
cell viability assay. FIG. 12B is a dose matrix showing excess over
Bliss for the BVD-523/dabrafenib combination. FIG. 12C and FIG. 12D
show % viability relative to DMSO only treated controls for
dabrafenib and BVD-523 single agent treatments in A375 cells using
the Alamar Blue cell viability assay. FIG. 12E shows % viability
relative to DMSO only treated controls for dabrafenib and BVD-523
combination treatments in A375 cells using the Alamar Blue cell
viability assay.
[0040] FIG. 13A is a dose matrix showing % inhibition of the
BVD-523/dabrafenib combination in A375 cells using the
CellTiter-Glo cell viability assay. FIG. 13B is a dose matrix
showing excess over Bliss for the BVD-523/dabrafenib combination.
FIG. 13C and FIG. 13D show % viability relative to DMSO only
treated controls for dabrafenib and BVD-523 single agent treatments
in A375 cells using the CellTiter-Glo cell viability assay. FIG.
13E shows % viability relative to DMSO only treated controls for
dabrafenib and BVD-523 combination treatments in A375 cells using
the CellTiter-Glo cell viability assay.
[0041] FIG. 14A is a dose matrix showing % inhibition of the
trametinib/BVD-523 combination in A375 cells using the Alamar Blue
cell viability assay. FIG. 14B is a dose matrix showing excess over
Bliss for the trametinib/BVD-523 combination. FIG. 14C and FIG. 14D
show % viability relative to DMSO only treated controls for BVD-523
and trametinib single agent treatments in A375 cells using the
Alamar Blue cell viability assay. FIG. 14E shows % viability
relative to DMSO only treated controls for BVD-523 and trametinib
combination treatments in A375 cells using the Alamar Blue cell
viability assay.
[0042] FIG. 15A is a dose matrix showing % inhibition of the
trametinib/BVD-523 combination in A375 cells using the
CellTiter-Glo cell viability assay. FIG. 15B is a dose matrix
showing excess over Bliss for the trametinib/BVD-523 combination.
FIG. 15C and FIG. 15D show % viability relative to DMSO only
treated controls for BVD-523 and trametinib single agent treatments
in A375 cells using the CellTiter-Glo cell viability assay. FIG.
15E shows % viability relative to DMSO only treated controls for
BVD-523 and trametinib combination treatments in A375 cells using
the CellTiter-Glo cell viability assay.
[0043] FIG. 16A-FIG. 16D are a set of images showing Western blot
analysis of MAPK signaling in A375 cells after a 4 hour treatment
with various concentrations (in nM) of BVD-523, dabrafenib (Dab),
and Trametinib (Tram). 40 .mu.g of total protein was loaded in each
lane except where indicated otherwise. In this experiment,
duplicate samples were collected. FIG. 16A and FIG. 16B show
results from duplicate samples. Similarly, FIG. 16C and FIG. 16D
also show results from duplicate samples. In FIG. 16A and FIG. 16B,
pRSK1 had a relatively weak signal in A375 cells compared to other
markers. A different pRSK1-S380 antibody from Cell Signaling (cat.
#11989) was tested but did not give a detectable signal (data not
shown). In FIG. 16C and FIG. 16D, pCRAF-338 gave a minimal
signal.
[0044] FIG. 17A-FIG. 17D are a set of images showing Western blot
analysis of MAPK signaling in a human colorectal carcinoma cell
line (HCT116 cells) after a 4 hour treatment with various
concentrations (in nM) of BVD-523, dabrafenib (Dab), and Trametinib
(Tram). 40 .mu.g of total protein was loaded in each lane except
where indicated otherwise. In this experiment, duplicate samples
were collected. FIG. 17A and FIG. 17B show results from duplicate
samples. Similarly, FIG. 17C and FIG. 17D also show results from
duplicate samples. In FIG. 17A and FIG. 17B, pRSK1 levels appear to
be very low in HCT116 cells, and in FIG. 17C and FIG. 17D,
pCRAF-338 signal was also very weak.
[0045] FIG. 18A-FIG. 18D are a set of images showing Western blot
analysis of cell cycle and apoptosis signaling in A375 melanoma
cells after a 24 hour treatment with various concentrations (in nM)
of BVD-523 ("BVD523"), trametinib ("tram") and/or dabrafenib
("Dab") as labelled. 50 .mu.g of total protein was loaded in each
lane except where indicated otherwise. In this experiment,
duplicate samples were collected. FIG. 18A and FIG. 18B show
results from duplicate samples. Similarly, FIG. 18C and FIG. 18D
also show results from duplicate samples. In FIG. 18A and FIG. 18B,
no band of a size corresponding to cleaved PARP (89 kDa) was
apparent.
[0046] FIG. 19 shows that BVD-523 can treat acquired resistance to
targeted drugs in-vivo. A patient-derived line, ST052C, was
isolated from a BRAFV600E melanoma patient that progressed
following 10 months of therapy with MAPK-pathway directed
therapies. Treated ex vivo, ST052C exhibited acquired
cross-resistance to dabrafenib at 50 mg/kg BID. Meanwhile, BVD-523
was effective in ST052C as a single-agent at 100 mg/kg BID.
[0047] FIG. 20 is a flowchart showing the dose escalation protocol
used herein.
[0048] FIG. 21 shows a schematic of the mitogen-activated protein
kinases (MAPK) pathway.
[0049] FIG. 22A-FIG. 22E show the results of single agent
proliferation assays. Proliferation results are shown for treatment
with BVD-523 (FIG. 22A), SCH772984 (FIG. 22B), Dabrafenib (FIG.
22C), Trametinib (FIG. 22D), and Paclitaxel (FIG. 22E).
[0050] FIG. 23A-FIG. 23O show the results of the combination of
BVD-523 and Dabrafenib. FIG. 23A shows a dose matrix showing
inhibition (%) for the combination in RKO parental cells. FIG.
23B-FIG. 23C show the results of single agent proliferation assays
for the combination in FIG. 23A. FIG. 23D shows Loewe excess for
the combination in FIG. 23A and FIG. 23E shows Bliss excess for the
combination in FIG. 23A. FIG. 23F shows a dose matrix showing
inhibition (%) for the combination in RKO MEK1 (Q56P/+)-clone 1
cells. FIG. 23G-FIG. 23H show the results of single agent
proliferation assays for the combination in FIG. 23F. FIG. 23I
shows Loewe excess for the combination in FIG. 23F and FIG. 23J
shows Bliss excess for the combination in FIG. 23F. FIG. 23K shows
a dose matrix showing inhibition (%) for the combination in RKO
MEK1 (Q56P/+)-clone 2 cells. FIG. 23L-FIG. 23M show the results of
single agent proliferation assays for the combination in FIG. 23K.
FIG. 23N shows Loewe excess for the combination in FIG. 23K and
FIG. 23O shows Bliss excess for the combination in FIG. 23K.
[0051] FIG. 24A-FIG. 24O show the results of the combination of
SCH772984 and Dabrafenib. FIG. 24A shows a dose matrix showing
inhibition (%) for the combination in RKO parental cells. FIG.
24B-FIG. 24C show the results of single agent proliferation assays
for the combination in FIG. 24A. FIG. 24D shows Loewe excess for
the combination in FIG. 24A and FIG. 24E shows Bliss excess for the
combination in FIG. 24A. FIG. 24F shows a dose matrix showing
inhibition (%) for the combination in RKO MEK1 (Q56P/+)-clone 1
cells. FIG. 24G-FIG. 24H show the results of single agent
proliferation assays for the combination in FIG. 24F. FIG. 24I
shows Loewe excess for the combination in FIG. 24F and FIG. 24J
shows Bliss excess for the combination in FIG. 24F. FIG. 24K shows
a dose matrix showing inhibition (%) for the combination in RKO
MEK1 (Q56P/+)-clone 2 cells. FIG. 24L-FIG. 24M show the results of
single agent proliferation assays for the combination in FIG. 24K.
FIG. 24N shows Loewe excess for the combination in FIG. 24K and
FIG. 24O shows Bliss excess for the combination in FIG. 24K.
[0052] FIG. 25A-FIG. 25O show the results of the combination of
Trametinib and Dabrafenib. FIG. 25A shows a dose matrix showing
inhibition (%) for the combination in RKO parental cells. FIG.
25B-FIG. 25C show the results of single agent proliferation assays
for the combination in FIG. 25A. FIG. 25D shows Loewe excess for
the combination in FIG. 25A and FIG. 25E shows Bliss excess for the
combination in FIG. 25A. FIG. 25F shows a dose matrix showing
inhibition (%) for the combination in RKO MEK1 (Q56P/+)-clone 1
cells. FIG. 25G-FIG. 25H show the results of single agent
proliferation assays for the combination in FIG. 25F. FIG. 25I
shows Loewe excess for the combination in FIG. 25F and FIG. 25J
shows Bliss excess for the combination in FIG. 25F. FIG. 25K shows
a dose matrix showing inhibition (%) for the combination in RKO
MEK1 (Q56P/+)-clone 2 cells. FIG. 25L-FIG. 25M show the results of
single agent proliferation assays for the combination in FIG. 25K.
FIG. 25N shows Loewe excess for the combination in FIG. 25K and
FIG. 25O shows Bliss excess for the combination in FIG. 25K.
[0053] FIG. 26A shows Lowe Volumes for the combinations tested.
FIG. 26B shows Bliss Volumes for the combinations tested. FIG. 26C
shows Synergy Scores for the combinations tested.
[0054] FIG. 27A-FIG. 27I show the changes in MAPK and Effector
Pathway Signaling in MEK acquired resistance. Isogenic RKO parental
and MEK1 (Q56P/+) cells were treated with compound for 4 or 24 h
and then immuno-blotted with the indicated antibodies. Dabrafenib
was the BRAF inhibitor and trametinib was the MEK inhibitor. FIG.
27A shows increased signaling in RKO MEK1 (Q56P/+) cells. FIG.
27B-FIG. 27C show the results of a 4 hour treatment in Experiment 1
(See, Example 7) in RKO Parental (27B) and RKO MEK1 (Q56P/+) (27C)
cells. FIG. 27D-FIG. 27E show the results of a 4 hour treatment in
Experiment 2 (See, Example 7) in RKO Parental (27D) and RKO MEK1
(Q56P/+) (27E) cells. FIG. 27F-FIG. 27G show the results of a 4
hour treatment in Experiment 2 (See, Example 7) in RKO Parental
(27F) and RKO MEK1 (Q56P/+) (27G) cells. FIG. 27H-FIG. 27I show a
summary of results in RKO Parental (27H) and RKO MEK1 (Q56P/+)
(27I) cells.
[0055] FIG. 28A-FIG. 28E show the results of the combination of
BVD-523 and SCH772984. FIG. 28A shows a dose matrix showing
inhibition (%) for the combination in A375 cells. FIG. 28B-FIG. 28C
show the results of single agent proliferation assays for the
combination in FIG. 28A. FIG. 28D shows Loewe excess for the
combination in FIG. 28A and FIG. 28E shows Bliss excess for the
combination in FIG. 28A.
[0056] FIG. 29A-FIG. 29F show discovery and characterization of the
novel ERK1/2 inhibitor BVD-523 (ulixertinib). FIG. 29A shows that
BVD-523 demonstrates inhibition in a reversible ATP-competitive
manner. This is demonstrated by a linear increase in IC.sub.50
values for inhibition of ERK2 with increasing ATP concentration as
shown in FIG. 29B. FIG. 29C shows a representative plot of the
dose-response curve and FIG. 29D shows a plot of IC.sub.50 over
time. FIG. 29E shows BVD-523 binding to ERK2 and phospho-ERK2
(pERK2), compared with negative control protein p38. FIG. 29F shows
BVD-523 binding to ERK2 compared with the ERK inhibitors SCH772984
and pyrazolylpyrrole.
[0057] FIG. 30A-FIG. 30D show that BVD 523 inhibits cellular
proliferation and enhances caspase 3 and caspase 7 activity in
vitro. FIG. 30A shows that BVD-523 demonstrates preferential
activity in cells with MAPK pathway mutations, as defined by the
presence of mutations in RAS family members and RAF. In addition,
as shown in FIG. 30B, BVD-523 blocks sensitive cell lines in the G1
phase of the cell cycle. FIG. 30C shows that BVD-523 induced a
concentration- and time-dependent increase in caspase activity in
the A375, WM266, and LS411N cancer cell lines after 72 hours of
exposure. FIG. 30D shows that the MAPK pathway and effector
proteins are modulated by acute (4-hour) and prolonged (24-hour)
BVD-523 treatment in BRAF.sup.V600E-mutant A375 cells.
[0058] FIG. 31A-FIG. 31C show in vivo BVD-523 anti-tumor activity.
BVD-523 monotherapy inhibits tumor growth in (FIG. 31A) A375 and
(FIG. 31B) Colo205 cell line xenograft models (.sup.aP<0.0001,
compared with vehicle control; CPT-11 dosed on Day 14 and Day 18
only). Abbreviations: BID, twice daily; CMC,
carboxymethylcellulose; QD, every day; Q4D, every 4 days. FIG. 31C
shows that in Colo205 xenografts, increased ERK1/2 phosphorylation
correlates with BVD-523 concentration.
[0059] FIG. 32A shows signaling effects of ERK1/2 inhibitors. Using
RPPA, effects on proteins are measured in cell lines (A375, AN3Ca,
Colo205, HCT116, HT29 and MIAPaca2) following treatment with ERK1/2
inhibitors BVD-523 (BVD), Vx11e (Vx), GDC-0994 (GDC), or SCH722984
(SCH). FIG. 32B shows that the ERK inhibitors BVD-523, GDC-0994,
and Vx11e have differential effects on phospho-ERK (ERK 1/2 T202
Y204) compared with SCH722984; phospho-RSK (p90 RSK 380) and Cyclin
D1 are inhibited by the ERK inhibitors tested. Abbreviations:
BRAFi, BRAF inhibitors; MEKi, MEK inhibitors. FIG. 32C shows a
western blot assay of cellular and nuclear fractions from a RKO
cell line following treatment with BVD-523, trametinib, SCH722984,
or dabrafenib. Histone H3 (nuclear localized protein) and HSP90
(cytoplasmically localized protein) were included as positive
controls to confirm that the nuclear and cytoplasmic fractions were
properly enriched; nuclear fractions have high H3 and cytoplasmic
fractions have higher HSP90.
[0060] FIG. 33 shows that the ERK inhibitors BVD-523, Vx11,
GDC-0994, and SCH772984 (SCH) demonstrate cell line-dependent
changes in phospho-ATK levels. Abbreviation: DMSO, dimethyl
sulfoxide.
[0061] FIG. 34A-FIG. 34D show that BVD-523 demonstrates activity in
models of resistance to BRAF/MEK inhibition. The appearance of
resistance to BVD-523, dabrafenib, or trametinib in BRAF.sup.V600E
A375 cells following exposure to increasing concentrations of drug
is indicated. A strict set of "criteria" was applied to determine
when the dose could be increased in order to ensure that the
kinetics of the acquisition of resistance between treatments was
comparable. See, Example 1. Time is shown against multipliers of
IC.sub.50; each point on the plotted line represents a change of
medium or cell split. FIG. 34A shows that adapting cells to growth
in the presence of BVD-523 was more challenging than with either
dabrafenib or trametinib. FIG. 34B shows that BVD-523 sensitivity
is retained in A375 cells cultured to acquire resistance to
combined BRAF (dabrafenib)+MEK (trametinib) inhibition. In FIG.
34C, cells were treated with compound for 96 h and viability was
assessed using CellTiter-Glo.RTM.. BVD-523 activity is retained in
BRAF.sup.V600E RKO cells cross-resistant to BRAF (dabrafenib) and
MEK (trametinib) inhibitors due to endogenous heterozygous knock-in
of MEK1.sup.Q56P. FIG. 34D shows that BVD-523 inhibition of pRSK in
BRAF.sup.V600E-mutant cell line RKO is maintained in the presence
of MEK1.sup.Q56P, which confers resistance to MEK and BRAF
inhibition. Knock-in of KRAS mutant alleles into SW48 cell lines
significantly diminishes sensitivity to the MEK inhibitors
trametinib and selumetinib, while comparatively sensitivity to
BVD-523 is retained.
[0062] FIG. 35A shows BVD-523 in vivo activity in xenografts
derived from a vemurafenib-relapsed patient. Mean tumor volume
(.+-.SEM) is shown for BVD-523 100 mg/kg BID alone, dabrafenib 50
mg/kg BID alone, and BVD-523 100 mg/kg BID plus dabrafenib 50 mg/kg
BID. Abbreviations: BID, twice daily; SEM, standard error of
mean.
[0063] FIG. 36A-FIG. 36D show the benefit of combined BVD-523 and
BRAF inhibition. FIG. 36A-FIG. 36B show that the combination of
BVD-523 plus dabrafenib exhibited superior antitumor activity
compared with treatment with either agent alone in a A375
BRAF.sup.V600E-mutant melanoma cell line xenograft model with a
tumor start volume of 75-144 mm.sup.3. FIG. 36C-FIG. 36D show
similar data from the same model with an enlarged tumor volume
(700-800 mm.sup.3) at the start of dosing. Plots of mean tumor
growth (left panels) and Kaplan-Meier survival (right panels) are
presented for each study. Abbreviations: BID, twice daily; QD, once
daily.
[0064] FIG. 37A shows that, in SW48 colorectal cells engineered
with KRAS alleles, response to paclitaxel was unaltered compared to
control. FIG. 37B shows combination interactions between BVD-523
and vemurafenib, which were assessed using an 8.times.10 matrix of
concentrations using the Loewe Additivity and Bliss Independence
Models, and analyzed with Horizon's Chalice, Bioinformatics
Software. Chalice enables potential synergistic interactions to be
identified by displaying the calculated excess inhibition over that
predicted as being additive across the dose matrix as a heat map,
and by reporting a quantitative "Synergy Score" based on the Loewe
model. The results suggest that interactions between BVD-523 and
vemurafenib are at least additive, and in some cases synergistic in
melanoma cell lines carrying a BRAF.sup.V600E mutation. FIG. 37C
shows that BVD-523 in combination with dabrafenib markedly delays
the onset of acquired resistance in A375 BRAF.sup.V600E melanoma
cells. The temporal acquisition of resistance in response to
escalating concentrations of dabrafenib alone or in combination
with BVD-523 or trametinib was assessed. Strict criteria were
applied as to when the dose could be increased to ensure that the
kinetics of adaptation was comparable between treatments. See,
Example 1.
[0065] FIG. 38 shows that BVD-523 inhibits ex vivo PMA-stimulated
RSK1/2 phosphorylation in human whole blood. Averages of BVD-523
concentration data set are indicated by (-). n=20 for each
concentration of BVD-523. Abbreviations: PBMC, peripheral blood
mononuclear cells; RSK, ribosomal S6 kinase.
[0066] FIG. 39A shows steady-state BVD-523 pharmacokinetics (Cycle
1, Day 15). The dashed red line indicates an EC.sub.50 200 ng/mL
HWB. Abbreviations: AUC, area under the curve; BID, twice daily;
C.sub.max, maximum concentration; EC.sub.50, 50% maximum effective
concentration; HWB, human whole blood; SD, standard deviation. FIG.
39B shows pharmacodynamic inhibition of ERK phosphorylation by
BVD-523 in human whole blood. Abbreviations: BID, twice daily;
pRSK, phospho-RSK; RSK, ribosomal S6 kinase.
[0067] FIG. 40A shows the best radiographic response in patients
treated with BVD-523. Included are all patients with disease
measured by RECIST v1.1 who received .gtoreq.1 dose of study
treatment and had >1 on-treatment tumor assessment (25/27; 2 did
not receive both scans of target lesions). Response was measured as
the change from baseline in the sum of the longest diameter of each
target lesion. Dose shown is that which the patient was receiving
at the time of response. The dashed line indicates the threshold
for a partial response according to RECIST v1.1. Abbreviations:
CRC, colorectal cancer; NET, neuroendocrine tumors; NSCLC,
non-small cell lung cancer; NSGCT, nonseminomatous germ cell
tumors; PNET, pancreatic NET; PTC, papillary thyroid cancer; RECIST
v1.1, Response Evaluation Criteria in Solid Tumors version 1.1;
SLD, sum of the largest diameter. FIG. 40B shows a computerized
tomography scan of a confirmed partial response in a 61-year-old
patient with a BRAF-mutant melanoma treated with BVD-523.
[0068] FIG. 41 shows tumor response and tumor progression. Shown is
a swimmer plot of tumor response, tumor progression, and duration
of treatment in response-evaluable patients treated with BVD-523.
Origin of the vertical axis corresponds to randomization date or
reference start date. Analysis cut-off date: Dec. 1, 2015.
Abbreviation: BID, twice daily.
DETAILED DESCRIPTION OF THE INVENTION
[0069] One embodiment of the present invention is a method for
treating or ameliorating the effects of a cancer in a subject,
which cancer is refractory or resistant to non-ERK MAPK pathway
inhibitor therapy. The method comprises administering to the
subject an effective amount of BVD-523 or a pharmaceutically
acceptable salt thereof.
[0070] As used herein, the terms "treat," "treating," "treatment"
and grammatical variations thereof mean subjecting an individual
subject to a protocol, regimen, process or remedy, in which it is
desired to obtain a physiologic response or outcome in that
subject, e.g., a patient. In particular, the methods and
compositions of the present invention may be used to slow the
development of disease symptoms or delay the onset of the disease
or condition, or halt the progression of disease development.
However, because every treated subject may not respond to a
particular treatment protocol, regimen, process or remedy, treating
does not require that the desired physiologic response or outcome
be achieved in each and every subject or subject population, e.g.,
patient population. Accordingly, a given subject or subject
population, e.g., patient population may fail to respond or respond
inadequately to treatment.
[0071] As used herein, the terms "ameliorate", "ameliorating" and
grammatical variations thereof mean to decrease the severity of the
symptoms of a disease in a subject.
[0072] As used herein, a "subject" is a mammal, preferably, a
human. In addition to humans, categories of mammals within the
scope of the present invention include, for example, farm animals,
domestic animals, laboratory animals, etc. Some examples of farm
animals include cows, pigs, horses, goats, etc. Some examples of
domestic animals include dogs, cats, etc. Some examples of
laboratory animals include primates, rats, mice, rabbits, guinea
pigs, etc.
[0073] In the present invention, BVD-523 corresponds to a compound
according to formula (I):
##STR00001##
and pharmaceutically acceptable salts thereof. BVD-523 may be
synthesized according to the methods disclosed, e.g., in U.S. Pat.
No. 7,354,939. Enantiomers and racemic mixtures of both enantiomers
of BVD-523 are also contemplated within the scope of the present
invention. BVD-523 is an ERK1/2 inhibitor with a mechanism of
action that is believed to be, e.g., unique and distinct from
certain other ERK1/2 inhibitors, such as SCH772984 and the
pyrimidinal structure used by Hatzivassiliou et al. (2012). For
example, other ERK1/2 inhibitors, such as SCH772984, inhibit
autophosphorylation of ERK (Morris et al., 2013), whereas BVD-523
allows for the autophosphorylation of ERK while still inhibiting
ERK. (See, e.g., FIG. 18).
[0074] As used herein, the words "resistant" and "refractory" are
used interchangeably. Being "resistant" to non-ERK MAPK pathway
inhibitor therapy treatments means that non-ERK MAPK inhibitors
have reduced efficacy in treating cancer.
[0075] As used herein, a "non-ERK MAPK inhibitor" means any
substance that reduces the activity, expression or phosphorylation
of proteins or other members of the MAPK pathway that results in a
reduction of cell growth or an increase in cell death, with the
exception of ERK1/2 inhibitors. As used herein, an "ERK1/2
inhibitor" means those substances that (i) directly interact with
ERK1 and/or ERK2, e.g., by binding to ERK1/2 and (ii) decrease the
expression or the activity of ERK1 and/or ERK2 protein kinases.
Therefore, inhibitors that act upstream of ERK1/2, such as MEK
inhibitors and RAF inhibitors, are not ERK1/2 inhibitors according
to the present invention (but they are non-ERK MAPK inhibitors).
Non-limiting examples of ERK1/2 inhibitors according to the present
invention include AEZS-131 (Aeterna Zentaris), AEZS-136 (Aeterna
Zentaris), BVD-523 (BioMed Valley Discoveries, Inc.), SCH-722984
(Merck & Co.), SCH-772984 (Merck & Co.), SCH-900353
(MK-8353) (Merck & Co.), pharmaceutically acceptable salts
thereof, and combinations thereof.
[0076] An overview of the mammalian MAPK cascades is shown in FIG.
21. The MAPK pathway is reviewed in e.g., Akinleye et al., 2013.
Briefly, with respect to the ERK1/2 module in FIG. 21 (light purple
box), the MAPK 1/2 signaling cascade is activated by ligand binding
to receptor tyrosine kinases (RTK). The activated receptors recruit
and phosphorylate adaptor proteins Grb2 and SOS, which then
interact with membrane-bound GTPase Ras and cause its activation.
In its activated GTP-bound form, Ras recruits and activates RAF
kinases (A-RAF, B-RAF, and C-RAF/RAF-1). The activated RAF kinases
activate MAPK 1/2 (MKK1/2), which in turn catalyzes the
phosphorylation of threonine and tyrosine residues in the
activation sequence Thr-Glu-Tyr of ERK1/2. With respect to the
JNK/p38 module (yellow box in FIG. 21), upstream kinases, MAP3Ks,
such as MEKK1/4, ASK1/2, and MLK1/2/3, activate MAP2K3/6 (MKK3/6),
MAP2K4 (MKK4), and MAP2K7 (MKK7). These MAP2K's then activate JNK
protein kinases, including JNK1, JNK2, and JNK3, as well as p38
.alpha./.beta./.gamma./.delta.. To execute their functions, JNKs
activate several transcription factors, including c-Jun, ATF-2,
NF-ATc1, HSF-1 and STAT3. With respect to the ERK5 module (blue box
in FIG. 21), the kinases upstream of MAP2K5 (MKK5) are MEKK2 and
MEKK3. The best characterized downstream target of MEK5 is ERK5,
also known as big MAP kinase 1 (BMK1) because it is twice the size
of other MAPKs.
[0077] Non-limiting examples of non-ERK MAPK pathway inhibitors
according to the present invention include RAS inhibitors, RAF
inhibitors (such as, e.g., inhibitors of A-RAF, B-RAF, C-RAF
(RAF-1)), MEK inhibitors, and combinations thereof. Preferably, the
non-ERK MAPK pathway inhibitors are BRAF inhibitors, MEK
inhibitors, and combinations thereof.
[0078] As used herein, a "RAS inhibitor" means those substances
that (i) directly interact with RAS, e.g., by binding to RAS and
(ii) decrease the expression or the activity of RAS. Non-limiting
exemplary RAS inhibitors include, but are not limited to, farnesyl
transferase inhibitors (such as, e.g., tipifarnib and lonafarnib),
farnesyl group-containing small molecules (such as, e.g., salirasib
and TLN-4601), DCAI, as disclosed by Maurer (Maurer et al., 2012),
Kobe0065 and and Kobe2602, as disclosed by Shima (Shima et al.,
2013), HBS 3 (Patgiri et al., 2011), and AIK-4 (Allinky).
[0079] As used herein, a "RAF inhibitor" means those substances
that (i) directly interact with RAF, e.g., by binding to RAF and
(ii) decrease the expression or the activity of RAF, such as, e.g.,
A-RAF, B-RAF, and C-RAF (RAF-1). Non-limiting exemplary RAF
inhibitors, including BRAF inhibitors, include:
##STR00002## ##STR00003## ##STR00004## ##STR00005##
##STR00006##
AAL881 (Novartis); AB-024 (Ambit Biosciences), ARQ-736 (ArQule),
ARQ-761 (ArQule), AZ628 (Axon Medchem BV), BeiGene-283 (BeiGene),
BIIB-024 (MLN 2480) (Sunesis & Takeda), b-raf inhibitor
(Sareum), BRAF kinase inhibitor (Selexagen Therapeutics), BRAF
siRNA 313 (tacaccagcaagctagatgca) and 523 (cctatcgttagagtcttcctg)
(Liu et al., 2007), CTT239065 (Institute of Cancer Research),
dabrafenib (GSK2118436), DP-4978 (Deciphera Pharmaceuticals),
HM-95573 (Hanmi), GDC-0879 (Genentech), GW-5074 (Sigma Aldrich),
ISIS 5132 (Novartis), L779450 (Merck), LBT613 (Novartis), LErafAON
(NeoPharm, Inc.), LGX-818 (Novartis), pazopanib (GlaxoSmithKline),
PLX3202 (Plexxikon), PLX4720 (Plexxikon), PLX5568 (Plexxikon),
RAF-265 (Novartis), RAF-365 (Novartis), regorafenib (Bayer
Healthcare Pharmaceuticals, Inc.), RO 5126766 (Hoffmann-La Roche),
SB-590885 (GlaxoSmithKline), SB699393 (GlaxoSmithKline), sorafenib
(Onyx Pharmaceuticals), TAK 632 (Takeda), TL-241 (Teligene),
vemurafenib (RG7204 or PLX4032) (Daiichi Sankyo), XL-281
(Exelixis), ZM-336372 (AstraZeneca), pharmaceutically acceptable
salts thereof, and combinations thereof.
[0080] As used herein, a "MEK inhibitor" means those substances
that (i) directly interact with MEK, e.g., by binding to MEK and
(ii) decrease the expression or the activity of MEK. Thus,
inhibitors that act upstream of MEK, such as RAS inhibitors and RAF
inhibitors, are not MEF inhibitors according to the present
invention. Non-limiting examples of MEK inhibitors include anthrax
toxin, antroquinonol (Golden Biotechnology), ARRY-142886
(6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-c-
arboxylic acid (2-hydroxy-ethoxy)-amide) (Array BioPharma),
ARRY-438162 (Array BioPharma), AS-1940477 (Astellas), AS-703988
(Merck KGaA), bentamapimod (Merck KGaA), BI-847325 (Boehringer
Ingelheim), E-6201 (Eisai), GDC-0623 (Hoffmann-La Roche), GDC-0973
(cobimetinib) (Hoffmann-La Roche), L783277 (Merck), lethal factor
portion of anthrax toxin, MEK162 (Array BioPharma), PD 098059
(2-(2'-amino-3'-methoxyphenyl)-oxanaphthalen-4-one) (Pfizer), PD
184352 (CI-1040) (Pfizer), PD-0325901 (Pfizer), pimasertib
(Santhera Pharmaceuticals), RDEA119 (Ardea Biosciences/Bayer),
refametinib (AstraZeneca), RG422 (Chugai Pharmaceutical Co.),
RO092210 (Roche), RO4987655 (Hoffmann-La Roche), RO5126766
(Hoffmann-La Roche), selumetinib (AZD6244) (AstraZeneca), SL327
(Sigma), TAK-733 (Takeda), trametinib (Japan Tobacco), U0126
(1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene)
(Sigma), WX-554 (Wilex), YopJ polypeptide (Mittal et al., 2010),
pharmaceutically acceptable salts thereof, and combinations
thereof.
[0081] In one aspect of this embodiment, substantially all
phosphorylation of ribosomal s6 kinase (RSK) is inhibited after
administration of BVD-523 or a pharmaceutically acceptable salt
thereof. As used herein in the context of RSK phosphorylation,
"substantially all" means a reduction of greater than 50%
reduction, preferably greater than 55%, 60%, 65%, 70%, 75%, 80%,
85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%
reduction.
[0082] In another aspect of this embodiment, the cancer has MAPK
activity. As used herein, having "MAPK activity" means that
proteins downstream of ERK are still active, even if proteins
upstream of ERK may not be active. Such a cancer may be a solid
tumor cancer or a hematologic cancer.
[0083] In the present invention, cancers include both solid and
hemotologic cancers. Non-limiting examples of solid cancers include
adrenocortical carcinoma, anal cancer, bladder cancer, bone cancer
(such as osteosarcoma), brain cancer, breast cancer, carcinoid
cancer, carcinoma, cervical cancer, colon cancer, endometrial
cancer, esophageal cancer, extrahepatic bile duct cancer, Ewing
family of cancers, extracranial germ cell cancer, eye cancer,
gallbladder cancer, gastric cancer, germ cell tumor, gestational
trophoblastic tumor, head and neck cancer, hypopharyngeal cancer,
islet cell carcinoma, kidney cancer, large intestine cancer,
laryngeal cancer, leukemia, lip and oral cavity cancer, liver
tumor/cancer, lung tumor/cancer, lymphoma, malignant mesothelioma,
Merkel cell carcinoma, mycosis fungoides, myelodysplastic syndrome,
myeloproliferative disorders, nasopharyngeal cancer, neuroblastoma,
oral cancer, oropharyngeal cancer, osteosarcoma, ovarian epithelial
cancer, ovarian germ cell cancer, pancreatic cancer, paranasal
sinus and nasal cavity cancer, parathyroid cancer, penile cancer,
pituitary cancer, plasma cell neoplasm, prostate cancer,
rhabdomyosarcoma, rectal cancer, renal cell cancer, transitional
cell cancer of the renal pelvis and ureter, salivary gland cancer,
Sezary syndrome, skin cancers (such as cutaneous t-cell lymphoma,
Kaposi's sarcoma, mast cell tumor, and melanoma), small intestine
cancer, soft tissue sarcoma, stomach cancer, testicular cancer,
thymoma, thyroid cancer, urethral cancer, uterine cancer, vaginal
cancer, vulvar cancer, and Wilms' tumor.
[0084] Examples of hematologic cancers include, but are not limited
to, leukemias, such as adult/childhood acute lymphoblastic
leukemia, adult/childhood acute myeloid leukemia, chronic
lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell
leukemia, lymphomas, such as AIDS-related lymphoma, cutaneous
T-cell lymphoma, adult/childhood Hodgkin lymphoma, mycosis
fungoides, adult/childhood non-Hodgkin lymphoma, primary central
nervous system lymphoma, Sezary syndrome, cutaneous T-cell
lymphoma, and Waldenstrom macroglobulinemia, as well as other
proliferative disorders such as chronic myeloproliferative
disorders, Langerhans cell histiocytosis, multiple myeloma/plasma
cell neoplasm, myelodysplastic syndromes, and
myelodysplastic/myeloproliferative neoplasms.
[0085] Preferably, the cancer is selected from the group consisting
of a cancer of the large intestine, breast cancer, pancreatic
cancer, skin cancer, and endometrial cancers. More preferably, the
cancer is melanoma.
[0086] In another aspect of this embodiment, the method further
comprises administering to the subject at least one additional
therapeutic agent effective for treating or ameliorating the
effects of the cancer. The additional therapeutic agent may be
selected from the group consisting of an antibody or fragment
thereof, a cytotoxic agent, a toxin, a radionuclide, an
immunomodulator, a photoactive therapeutic agent, a
radiosensitizing agent, a hormone, an anti-angiogenesis agent, and
combinations thereof.
[0087] As used herein, an "antibody" encompasses naturally
occurring immunoglobulins as well as non-naturally occurring
immunoglobulins, including, for example, single chain antibodies,
chimeric antibodies (e.g., humanized murine antibodies), and
heteroconjugate antibodies (e.g., bispecific antibodies). Fragments
of antibodies include those that bind antigen, (e.g., Fab',
F(ab').sub.2, Fab, Fv, and rIgG). See also, e.g., Pierce Catalog
and Handbook, 1994-1995 (Pierce Chemical Co., Rockford, Ill.);
Kuby, J., Immunology, 3rd Ed., W.H. Freeman & Co., New York
(1998). The term antibody also includes bivalent or bispecific
molecules, diabodies, triabodies, and tetrabodies. The term
"antibody" further includes both polyclonal and monoclonal
antibodies.
[0088] Examples of therapeutic antibodies that may be used in the
present invention include rituximab (Rituxan), Cetuximab (Erbitux),
bevacizumab (Avastin), and Ibritumomab (Zevalin).
[0089] Cytotoxic agents according to the present invention include
DNA damaging agents, antimetabolites, anti-microtubule agents,
antibiotic agents, etc. DNA damaging agents include alkylating
agents, platinum-based agents, intercalating agents, and inhibitors
of DNA replication. Non-limiting examples of DNA alkylating agents
include cyclophosphamide, mechlorethamine, uramustine, melphalan,
chlorambucil, ifosfamide, carmustine, lomustine, streptozocin,
busulfan, temozolomide, pharmaceutically acceptable salts thereof,
prodrugs, and combinations thereof. Non-limiting examples of
platinum-based agents include cisplatin, carboplatin, oxaliplatin,
nedaplatin, satraplatin, triplatin tetranitrate, pharmaceutically
acceptable salts thereof, prodrugs, and combinations thereof.
Non-limiting examples of intercalating agents include doxorubicin,
daunorubicin, idarubicin, mitoxantrone, pharmaceutically acceptable
salts thereof, prodrugs, and combinations thereof. Non-limiting
examples of inhibitors of DNA replication include irinotecan,
topotecan, amsacrine, etoposide, etoposide phosphate, teniposide,
pharmaceutically acceptable salts thereof, prodrugs, and
combinations thereof. Antimetabolites include folate antagonists
such as methotrexate and premetrexed, purine antagonists such as
6-mercaptopurine, dacarbazine, and fludarabine, and pyrimidine
antagonists such as 5-fluorouracil, arabinosylcytosine,
capecitabine, gemcitabine, decitabine, pharmaceutically acceptable
salts thereof, prodrugs, and combinations thereof. Anti-microtubule
agents include without limitation vinca alkaloids, paclitaxel
(Taxol.RTM.), docetaxel (Taxotere.RTM.), and ixabepilone
(Ixempra.RTM.). Antibiotic agents include without limitation
actinomycin, anthracyclines, valrubicin, epirubicin, bleomycin,
plicamycin, mitomycin, pharmaceutically acceptable salts thereof,
prodrugs, and combinations thereof.
[0090] Cytotoxic agents according to the present invention also
include an inhibitor of the PI3K/Akt pathway. Non-limiting examples
of an inhibitor of the PI3K/Akt pathway include A-674563 (CAS
#552325-73-2), AGL 2263, AMG-319 (Amgen, Thousand Oaks, Calif.),
AS-041164
(5-benzo[1,3]dioxol-5-ylmethylene-thiazolidine-2,4-dione),
AS-604850
(5-(2,2-Difluoro-benzo[1,3]dioxol-5-ylmethylene)-thiazolidine-2,4-dione),
AS-605240 (5-quinoxilin-6-methylene-1,3-thiazolidine-2,4-dione),
AT7867 (CAS #857531-00-1), benzimidazole series, Genentech (Roche
Holdings Inc., South San Francisco, Calif.), BML-257 (CAS
#32387-96-5), CAL-120 (Gilead Sciences, Foster City, Calif.),
CAL-129 (Gilead Sciences), CAL-130 (Gilead Sciences), CAL-253
(Gilead Sciences), CAL-263 (Gilead Sciences), CAS #612847-09-3, CAS
#681281-88-9, CAS #75747-14-7, CAS #925681-41-0, CAS #98510-80-6,
CCT128930 (CAS #885499-61-6), CH5132799 (CAS #1007207-67-1),
CHR-4432 (Chroma Therapeutics, Ltd., Abingdon, UK), FPA 124 (CAS
#902779-59-3), GS-1101 (CAL-101) (Gilead Sciences), GSK 690693 (CAS
#937174-76-0), H-89 (CAS #127243-85-0), Honokiol, IC87114 (Gilead
Science), IPI-145 (Intellikine Inc.), KAR-4139 (Karus Therapeutics,
Chilworth, UK), KAR-4141 (Karus Therapeutics), KIN-1 (Karus
Therapeutics), KT 5720 (CAS #108068-98-0), Miltefosine, MK-2206
dihydrochloride (CAS #1032350-13-2), ML-9 (CAS #105637-50-1),
Naltrindole Hydrochloride, OXY-111A (NormOxys Inc., Brighton,
Mass.), perifosine, PHT-427 (CAS #1191951-57-1), PI3 kinase delta
inhibitor, Merck KGaA (Merck & Co., Whitehouse Station, N.J.),
PI3 kinase delta inhibitors, Genentech (Roche Holdings Inc.), PI3
kinase delta inhibitors, Incozen (Incozen Therapeutics, Pvt. Ltd.,
Hydrabad, India), PI3 kinase delta inhibitors-2, Incozen (Incozen
Therapeutics), PI3 kinase inhibitor, Roche-4 (Roche Holdings Inc.),
PI3 kinase inhibitors, Roche (Roche Holdings Inc.), PI3 kinase
inhibitors, Roche-5 (Roche Holdings Inc.), PI3-alpha/delta
inhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd., South
San Francisco, Calif.), PI3-delta inhibitors, Cellzome (Cellzome
AG, Heidelberg, Germany), PI3-delta inhibitors, Intellikine
(Intellikine Inc., La Jolla, Calif.), PI3-delta inhibitors, Pathway
Therapeutics-1 (Pathway Therapeutics Ltd.), PI3-delta inhibitors,
Pathway Therapeutics-2 (Pathway Therapeutics Ltd.), PI3-delta/gamma
inhibitors, Cellzome (Cellzome AG), PI3-delta/gamma inhibitors,
Cellzome (Cellzome AG), PI3-delta/gamma inhibitors, Intellikine
(Intellikine Inc.), PI3-delta/gamma inhibitors, Intellikine
(Intellikine Inc.), PI3-delta/gamma inhibitors, Pathway
Therapeutics (Pathway Therapeutics Ltd.), PI3-delta/gamma
inhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd.),
PI3-gamma inhibitor Evotec (Evotec), PI3-gamma inhibitor, Cellzome
(Cellzome AG), PI3-gamma inhibitors, Pathway Therapeutics (Pathway
Therapeutics Ltd.), PI3K delta/gamma inhibitors, Intellikine-1
(Intellikine Inc.), PI3K delta/gamma inhibitors, Intellikine-1
(Intellikine Inc.), pictilisib (Roche Holdings Inc.), PIK-90 (CAS
#677338-12-4), SC-103980 (Pfizer, New York, N.Y.), SF-1126
(Semafore Pharmaceuticals, Indianapolis, Ind.), SH-5, SH-6,
Tetrahydro Curcumin, TG100-115 (Targegen Inc., San Diego, Calif.),
Triciribine, X-339 (Xcovery, West Palm Beach, Fla.), XL-499
(Evotech, Hamburg, Germany), pharmaceutically acceptable salts
thereof, and combinations thereof.
[0091] In the present invention, the term "toxin" means an
antigenic poison or venom of plant or animal origin. An example is
diphtheria toxin or portions thereof.
[0092] In the present invention, the term "radionuclide" means a
radioactive substance administered to the patient, e.g.,
intravenously or orally, after which it penetrates via the
patient's normal metabolism into the target organ or tissue, where
it delivers local radiation for a short time. Examples of
radionuclides include, but are not limited to, I-125, At-211,
Lu-177, Cu-67, I-131, Sm-153, Re-186, P-32, Re-188, In-114m, and
Y-90.
[0093] In the present invention, the term "immunomodulator" means a
substance that alters the immune response by augmenting or reducing
the ability of the immune system to produce antibodies or
sensitized cells that recognize and react with the antigen that
initiated their production. Immunomodulators may be recombinant,
synthetic, or natural preparations and include cytokines,
corticosteroids, cytotoxic agents, thymosin, and immunoglobulins.
Some immunomodulators are naturally present in the body, and
certain of these are available in pharmacologic preparations.
Examples of immunomodulators include, but are not limited to,
granulocyte colony-stimulating factor (G-CSF), interferons,
imiquimod and cellular membrane fractions from bacteria, IL-2,
IL-7, IL-12, CCL3, CCL26, CXCL7, and synthetic cytosine
phosphate-guanosine (CpG).
[0094] In the present invention, the term "photoactive therapeutic
agent" means compounds and compositions that become active upon
exposure to light. Certain examples of photoactive therapeutic
agents are disclosed, e.g., in U.S. Patent Application Serial No.
2011/0152230 A1, "Photoactive Metal Nitrosyls For Blood Pressure
Regulation And Cancer Therapy."
[0095] In the present invention, the term "radiosensitizing agent"
means a compound that makes tumor cells more sensitive to radiation
therapy. Examples of radiosensitizing agents include misonidazole,
metronidazole, tirapazamine, and trans sodium crocetinate.
[0096] In the present invention, the term "hormone" means a
substance released by cells in one part of a body that affects
cells in another part of the body. Examples of hormones include,
but are not limited to, prostaglandins, leukotrienes, prostacyclin,
thromboxane, amylin, antimullerian hormone, adiponectin,
adrenocorticotropic hormone, angiotensinogen, angiotensin,
vasopressin, atriopeptin, brain natriuretic peptide, calcitonin,
cholecystokinin, corticotropin-releasing hormone, encephalin,
endothelin, erythropoietin, follicle-stimulating hormone, galanin,
gastrin, ghrelin, glucagon, gonadotropin-releasing hormone, growth
hormone-releasing hormone, human chorionic gonadotropin, human
placental lactogen, growth hormone, inhibin, insulin, somatomedin,
leptin, liptropin, luteinizing hormone, melanocyte stimulating
hormone, motilin, orexin, oxytocin, pancreatic polypeptide,
parathyroid hormone, prolactin, prolactin releasing hormone,
relaxin, renin, secretin, somatostain, thrombopoietin,
thyroid-stimulating hormone, testosterone, dehydroepiandrosterone,
androstenedione, dihydrotestosterone, aldosterone, estradiol,
estrone, estriol, cortisol, progesterone, calcitriol, and
calcidiol.
[0097] Some compounds interfere with the activity of certain
hormones or stop the production of certain hormones. These
hormone-interfering compounds include, but are not limited to,
tamoxifen (Nolvadex.RTM.), anastrozole (Arimidex.RTM.), letrozole
(Femara.RTM.), and fulvestrant (Faslodex.RTM.). Such compounds are
also within the meaning of hormone in the present invention.
[0098] As used herein, an "anti-angiogenesis" agent means a
substance that reduces or inhibits the growth of new blood vessels,
such as, e.g., an inhibitor of vascular endothelial growth factor
(VEGF) and an inhibitor of endothelial cell migration.
Anti-angiogenesis agents include without limitation
2-methoxyestradiol, angiostatin, bevacizumab, cartilage-derived
angiogenesis inhibitory factor, endostatin, IFN-.alpha., IL-12,
itraconazole, linomide, platelet factor-4, prolactin, SU5416,
suramin, tasquinimod, tecogalan, tetrathiomolybdate, thalidomide,
thrombospondin, thrombospondin, TNP-470, ziv-aflibercept,
pharmaceutically acceptable salts thereof, prodrugs, and
combinations thereof.
[0099] Another embodiment of the present invention is a method for
treating or ameliorating the effects of a cancer in a subject. The
method comprises:
[0100] (a) identifying a subject with cancer that has become
refractory or resistant to BRAF inhibitor therapy, MEK inhibitor
therapy, or BRAF and MEK inhibitor therapy; and
[0101] (b) administering to the subject with said refractory or
resistant cancer an effective amount of an ERK inhibitor, which is
BVD-523 or a pharmaceutically acceptable salt thereof.
[0102] Suitable and preferred subjects are as disclosed herein. In
this embodiment, the methods may be used to treat the cancers
disclosed above. In accordance with the present invention, the
cancer may have MAPK activity.
[0103] In one aspect of this embodiment, identifying a subject with
cancer that is refractory or resistant to BRAF and/or MEK inhibitor
therapy comprises:
[0104] (a) obtaining a biological sample from the subject; and
[0105] (b) screening the sample to determine whether the subject
has become resistant to an inhibitor therapy selected from the
group consisting of BRAF inhibitor therapy, MEK inhibitor therapy,
and combinations thereof.
[0106] In the present invention, biological samples include, but
are not limited to, blood, plasma, urine, skin, saliva, and
biopsies. Biological samples are obtained from a subject by routine
procedures and methods which are known in the art.
[0107] Preferably, screening for a cancer that is refractory or
resistant to BRAF inhibitor therapy may comprise, e.g., identifying
(i) a switch between RAF isoforms, (ii) upregulation of RTK or NRAS
signaling, (iii) reactivation of mitogen activated protein kinase
(MAPK) signaling, (iv) the presence of a MEK activating mutation,
and combinations thereof.
[0108] A switch between RAF isoforms may occur in subjects having
acquired resistance to BRAF inhibitor therapy. To detect such a
switch, BRAF inhibitor-resistant tumor cells may be retrieved from
a patient and analyzed via Western blotting for ERK and phospho-ERK
levels in the presence of a BRAF inhibitor. Comparison with BRAF
inhibitor-sensitive cells treated with a BRAF inhibitor may reveal
higher levels of phospho-ERK in BRAF inhibitor-resistant tumor
cells, implying that a switch has taken place in which another RAF
isoform phosphorylates ERK in place of BRAF. Confirmation of which
RAF isoform has taken over may involve sh/siRNA-mediated knockdown
of ARAF and CRAF individually in BRAF inhibitor-resistant cells
exposed to a BRAF inhibitor, followed by subsequent Western
blotting for ERK and phospho-ERK levels. If, for example, ARAF
knockdown in BRAF inhibitor-resistant cells exposed to a BRAF
inhibitor still results in high levels of phospho-ERK, it would
indicate that CRAF has taken over phosphorylating ERK. Likewise, if
CRAF was knocked down in BRAF inhibitor-resistant cells exposed to
BRAF inhibitor and ERK was still highly phosphorylated, it would
mean that ARAF has taken over ERK phosphorylation. RAF isoform
switching may also involve simultaneous knockdown of ARAF and CRAF
in BRAF inhibitor-resistant cells in the presence of BRAF
inhibitor, effectively blocking all RAF-mediated phosphorylation. A
resulting decrease in ERK phosphorylation would indicate that the
BRAF inhibitor-resistant cells have the capacity to switch between
RAF isoforms in order to phosphorylate ERK (Villanueva, et al.,
2010).
[0109] Upregulation of RTK or NRAS signaling may also be a cause of
BRAF inhibitor resistance. Detection may, e.g., first involve using
Western blotting protocols with phospho-specific antibodies to
analyze the activation of the downstream RAF effectors MEK1/2 and
ERK1/2. If BRAF inhibitor-resistant cells show high activation
levels of these proteins in the presence of a BRAF inhibitor, RTK
or NRAS upregulation may be the cause. Gene expression profiling
(or other related methods) of BRAF inhibitor-resistant cells in the
presence of a BRAF inhibitor may reveal higher expression levels of
KIT, MET, EGFR, and PDGFR.beta. RTKs as compared to BRAF
inhibitor-sensitive cells. Real-time quantitative polymerase chain
reaction experiments, or other similar procedures, focusing on any
of these genes may confirm higher expression levels while
phospho-RTK arrays (R&D Systems, Minneapolis, Minn.) may show
elevated activation-associated tyrosine phosphorylation.
Alternatively, NRAS activation may be detected by various gene
sequencing protocols. Activating mutations in NRAS, particularly
Q61K, may indicate that B-RAF signaling has been bypassed. In
melanoma cells, activated NRAS uses C-RAF to signal to MEK-ERK.
Thus, activated NRAS may enable a similar bypass pathway in BRAF
inhibitor-resistant cells exposed to BRAF inhibitor. Further
confirmation of these mechanisms in a given BRAF
inhibitor-resistant sample may be accomplished, for example, using
sh/siRNA-mediated knockdown of upregulated RTKs or activated NRAS
in the presence of BRAF inhibitor. Any significant levels of growth
inhibition may indicate that upregulation of RTK or NRAS signaling
is the cause of BRAF inhibition in that particular sample
(Nazarian, et al., 2010).
[0110] Detecting reactivation of MAPK signaling in BRAF
inhibitor-resistant cells may indicate another bypass mechanism for
BRAF inhibitor resistance. COT and C-RAF have been shown to be
upregulated in a BRAF V600E background exposed to BRAF inhibitor.
Quantitative real-time RT-PCR, e.g., may reveal increased COT
expression in BRAF inhibitor-resistant cells in the presence of
BRAF inhibitor. Furthermore, sh/siRNA-mediated knockdown of COT in
BRAF inhibitor-resistant cells in the presence of BRAF inhibitor
may reduce the viability of BRAF inhibitor-resistant cells,
indicating that these particular cells may be sensitive to COT
inhibition and/or combination BRAF inhibitor/MEK inhibitor
treatments (Johannessen, et al., 2010).
[0111] Reactivation of MAPK signaling may also be accomplished in a
BRAF inhibitor-resistant background by activating mutations in
MEK1. Targeted, massively parallel sequencing of genomic DNA from a
BRAF inhibitor-resistant tumor may reveal activating mutations in
MEK1, such as C121S, G128D, N122D, and Y130, among others. Other,
undocumented mutations in MEK1 may be analyzed by, for example,
expressing the particular mutation in a BRAF inhibitor-sensitive
cell line such as A375. Determining levels of growth inhibition in
these cells upon exposure to BRAF inhibitor may indicate if the
MEK1 mutation is causing resistance to BRAF inhibitory therapy. To
confirm such a finding, Western blotting for elevated levels of
phospho-ERK1/2 in cells ectopically expressing the MEK1 mutation
may indicate that the MEK1 mutation is allowing the BRAF
inhibitor-resistant tumor to bypass BRAF and promote
phosphorylation of ERK through MEK1 (Wagle, et al., 2011).
[0112] In accordance with the present invention, screening for a
cancer that is refractory or resistant to MEK inhibitor therapy may
comprise, e.g., identifying (i) amplification of mutant BRAF, (ii)
STAT3 upregulation, (iii) mutations in the allosteric pocket of MEK
that directly block binding of inhibitors to MEK or lead to
constitutive MEK activity, and combinations thereof.
[0113] Amplification of mutant BRAF may cause MEK inhibitor
resistance. MEK inhibitor resistance is typically associated with
high levels of phosphorylated ERK and MEK in the presence of a MEK
inhibitor, which may be assessed via, for example, Western
blotting. Amplification of mutant BRAF in MEK inhibitor-resistant
cell lines may be detected by, for example, fluorescence in situ
hybridization (FISH) or quantitative PCR from genomic DNA of the
resistant cell lines. Confirmation that BRAF amplification is a
primary cause of MEK inhibitor resistance may entail using
BRAF-targeted sh/siRNAs in resistant cells. If a significant
decrease in MEK or ERK phosphorylation is observed, BRAF
amplification may be a suitable target for further therapeutic
approaches. (Corcoran, et al., 2010).
[0114] Identifying STAT3 upregulation may indicate that a
particular tumor sample is resistant to MEK inhibitor therapy.
Genome-wide expression profiling may reveal the STAT3 pathway to be
upregulated in a tumor. Other techniques, such as Western blotting
for phospho-STAT3 and real-time qPCR for the STAT
pathway-associated genes JAK1 and IL6ST may reveal upregulated
STAT3. Further confirmation that STAT3 upregulation causes MEK
inhibitor resistance in a particular sample may comprise the use of
sh/siRNAs against STAT3 in the sample followed by appropriate
Western blotting for MEK and ERK activation as well as
phospho-STAT3 and total STAT3. Growth inhibition studies may show
that STAT3 knockdown sensitizes previously MEK inhibitor-resistant
cells to MEK inhibition. A similar effect may be seen if the sample
were exposed to a STAT3 inhibitor such as JSI-124. Additional
confirmation that STAT3 upregulation is the cause of MEK inhibitor
resistance in a particular tumor could arise from Western blotting
for BIM expression, including BIM-EL, BIM-L, and BIM-SL. BIM
expression leads to MEK inhibitor-induced apoptosis, thus STAT3
upregulation may lower BIM levels. STAT3 is known to regulate the
expression of miR 17-92, which suppresses BIM expression.
Upregulated STAT3 may lead to higher levels of miR 17-92, which
will lower BIM levels and promote resistance to MEK inhibition.
Thus, real-time qPCR of miR 17-92 levels may also assist in
assessing whether STAT3 upregulation is causing MEK inhibition
resistance in a particular sample. (Dai, et al., 2011).
[0115] Mutations in the allosteric pocket of MEK that can directly
block binding of inhibitors to MEK or lead to constitutive MEK
activity may be detected by methods disclosed below. Such mutations
have been identified previously by Emery and colleagues (Emery, et
al., 2009) as well as Wang and colleagues (Wang et al., 2011).
Other mutations may affect MEK1 codons located within or abutting
the N-terminal negative regulatory helix, such as P124L and Q56P.
(Id.).
[0116] Methods for identifying mutations in nucleic acids, such as
the above identified MEK genes, are known in the art. Nucleic acids
may be obtained from biological samples. In the present invention,
biological samples include, but are not limited to, blood, plasma,
urine, skin, saliva, and biopsies. Biological samples are obtained
from a subject by routine procedures and methods which are known in
the art.
[0117] Non-limiting examples of methods for identifying mutations
include PCR, sequencing, hybrid capture, in-solution capture,
molecular inversion probes, fluorescent in situ hybridization
(FISH) assays, and combinations thereof.
[0118] Various sequencing methods are known in the art. These
include, but are not limited to, Sanger sequencing (also referred
to as dideoxy sequencing) and various sequencing-by-synthesis (SBS)
methods as disclosed in, e.g., Metzker 2005, sequencing by
hybridization, by ligation (for example, WO 2005021786), by
degradation (for example, U.S. Pat. Nos. 5,622,824 and 6,140,053)
and nanopore sequencing (which is commercially available from
Oxford Nanopore Technologies, UK). In deep sequencing techniques, a
given nucleotide in the sequence is read more than once during the
sequencing process. Deep sequencing techniques are disclosed in
e.g., U.S. Patent Publication No. 20120264632 and International
Patent Publication No. WO2012125848.
[0119] PCR-based methods for detecting mutations are known in the
art and employ PCR amplification, where each target sequence in the
sample has a corresponding pair of unique, sequence-specific
primers. For example, the polymerase chain reaction-restriction
fragment length polymorphism (PCR-RFLP) method allows for rapid
detection of mutations after the genomic sequences are amplified by
PCR. The mutation is discriminated by digestion with specific
restriction endonucleases and is identified by electrophoresis.
See, e.g., Ota et al., 2007. Mutations may also be detected using
real time PCR. See, e.g., International Application publication No.
WO2012046981.
[0120] Hybrid capture methods are known in the art and are
disclosed in e.g., U.S. Patent Publication No. 20130203632 and U.S.
Pat. Nos. 8,389,219 and 8,288,520. These methods are based on the
selective hybridization of the target genomic regions to
user-designed oligonucleotides. The hybridization can be to
oligonucleotides immobilized on high or low density microarrays
(on-array capture), or solution-phase hybridization to
oligonucleotides modified with a ligand (e.g. biotin) which can
subsequently be immobilized to a solid surface, such as a bead
(in-solution capture).
[0121] Molecular Inversion Probe (MIP) techniques are known in the
art and are disclosed in e.g., Absalan et al., 2008. This method
uses MIP molecules, which are special "padlock" probes (Nilsson et
al, 1994) for genotyping. A MIP molecule is a linear
oligonucleotide that contains specific regions, universal
sequences, restriction sites and a Tag (index) sequence (16-22 bp).
A MIP hybridizes directly around the genetic marker/SNP of
interest. The MIP method may also use a number of "padlock" probe
sets that hybridize to genomic DNA in parallel (Hardenbol et al.,
2003). In case of a perfect match, genomic homology regions are
ligated by undergoing an inversion in configuration (as suggested
by the name of the technique) and creating a circular molecule.
After the first restriction, all molecules are amplified with
universal primers. Amplicons are restricted again to ensure short
fragments for hybridization on a microarray. Generated short
fragments are labeled and, through a Tag sequence, hybridized to a
cTag (complementary strand for index) on an array. After the
formation of Tag-cTag duplex, a signal is detected.
[0122] The following Tables 1, 2, and 3 show the SEQ ID Nos. of
representative nucleic acid and amino acid sequences of wild type
BRAF, N-RAS, and MEK1 from various animals in the sequence listing.
These sequences may be used in methods for identifying subjects
with mutant BRAF, N-RAS, and MEK1 genotypes.
TABLE-US-00001 TABLE 1 BRAF sequences polypeptide or nucleic acid
Other SEQ ID NO. sequence Organism information 1 nucleic acid human
2 polypeptide human 3 nucleic acid rat (Rattus norvegicus) 4
polypeptide rat (Rattus norvegicus) 5 nucleic acid mouse, Mus
musculus 6 polypeptide mouse, Mus musculus 7 nucleic acid rabbit,
Oryctolagus cuniculus 8 polypeptide rabbit, Oryctolagus cuniculus 9
nucleic acid guinea pig, Cavia porcellus 10 polypeptide guinea pig,
Cavia porcellus 11 nucleic acid dog, Canis lupus variant x1
familiaris 12 polypeptide dog, Canis lupus variant x1 familiaris 13
nucleic acid dog, Canis lupus variant x2 familiaris 14 polypeptide
dog, Canis lupus variant x2 familiaris 15 nucleic acid cat, Felis
catus 16 polypeptide cat, Felis catus 17 nucleic acid cow, Bos
taurus variant X1 18 polypeptide cow, Bos taurus variant X1 19
nucleic acid cow, Bos taurus variant X2 20 polypeptide cow, Bos
taurus variant X2 21 nucleic acid cow, Bos taurus variant X3 22
polypeptide cow, Bos taurus variant X3 23 nucleic acid cow, Bos
taurus variant X4 24 polypeptide cow, Bos taurus variant X4 25
nucleic acid cow, Bos taurus variant X5 26 polypeptide cow, Bos
taurus variant X5 27 nucleic acid cow, Bos taurus variant X6 28
polypeptide cow, Bos taurus variant X6 29 nucleic acid cow, Bos
taurus variant X7 30 polypeptide cow, Bos taurus variant X7 31
nucleic acid cow, Bos taurus variant X8 32 polypeptide cow, Bos
taurus variant X8 33 nucleic acid cow, Bos taurus variant X9 34
polypeptide cow, Bos taurus variant X9 35 nucleic acid cow, Bos
taurus variant X10 36 polypeptide cow, Bos taurus variant X10 37
nucleic acid cow, Bos taurus variant X11 38 polypeptide cow, Bos
taurus variant X11 39 nucleic acid cow, Bos taurus variant 2 40
polypeptide cow, Bos taurus variant 2 41 nucleic acid horse, Equus
caballus 42 polypeptide horse, Equus caballus 43 nucleic acid
chicken, Gallus gallus 44 polypeptide chicken, Gallus gallus
TABLE-US-00002 TABLE 2 N-RAS sequences polypeptide or nucleic acid
Other SEQ ID NO. sequence Organism information 45 nucleic acid
human 46 polypeptide human 47 nucleic acid rat (Rattus norvegicus)
48 polypeptide rat (Rattus norvegicus) 49 nucleic acid mouse, Mus
musculus 50 polypeptide mouse, Mus musculus 51 nucleic acid guinea
pig, Cavia porcellus 52 polypeptide guinea pig, Cavia porcellus 53
nucleic acid guinea pig, Cavia porcellus variant X1 54 polypeptide
guinea pig, Cavia porcellus variant X1 55 nucleic acid dog, Canis
lupus familiaris 56 polypeptide dog, Canis lupus familiaris 57
nucleic acid cat, Felis catus 58 polypeptide cat, Felis catus 59
nucleic acid cow, Bos taurus 60 polypeptide cow, Bos taurus 61
nucleic acid chicken, Gallus gallus 62 polypeptide chicken, Gallus
gallus
TABLE-US-00003 TABLE 3 MEK1 sequences polypeptide or nucleic acid
SEQ ID NO. sequence Organism 63 nucleic acid human 64 polypeptide
human 65 nucleic acid rat (Rattus norvegicus) 66 polypeptide rat
(Rattus norvegicus) 67 nucleic acid mouse, Mus musculus 68
polypeptide mouse, Mus musculus 69 nucleic acid rabbit, Oryctolagus
cuniculus 70 polypeptide rabbit, Oryctolagus cuniculus 71 nucleic
acid guinea pig, Cavia porcellus 72 polypeptide guinea pig, Cavia
porcellus 73 nucleic acid dog, Canis lupus familiaris 74
polypeptide dog, Canis lupus familiaris 75 nucleic acid cat, Felis
catus 76 polypeptide cat, Felis catus 77 nucleic acid cow, Bos
taurus 78 polypeptide cow, Bos taurus 79 nucleic acid horse, Equus
caballus 80 polypeptide horse, Equus caballus 81 nucleic acid
chicken, Gallus gallus 82 polypeptide chicken, Gallus gallus
[0123] In another aspect of this embodiment, the method further
comprises administering at least one additional therapeutic agent,
preferably an inhibitor of the PI3K/Akt pathway, as disclosed
herein.
[0124] A further embodiment of the present invention is a method
for treating or ameliorating the effects of cancer in a subject,
which cancer is refractory or resistant to BRAF inhibitor therapy,
MEK inhibitor therapy, or both. The method comprises administering
to the subject an effective amount of BVD-523 or a pharmaceutically
acceptable salt thereof.
[0125] Suitable and preferred subjects are as disclosed herein. In
this embodiment, the methods may be used to treat the cancers
disclosed above, including those cancers with the mutational
backgrounds, resistance profiles, and MAPK activity identified
above. Methods of identifying such mutations are also as set forth
above.
[0126] In a further aspect of this embodiment, the method further
comprises administering to the subject at least one additional
therapeutic agent, preferably an inhibitor of the PI3K/Akt pathway,
as disclosed herein.
[0127] Another embodiment of the present invention is a method for
identifying a subject having cancer who would benefit from therapy
with an ERK inhibitor. The method comprises: [0128] (a) obtaining a
biological sample from the subject; and [0129] (b) screening the
sample to determine whether the subject has one or more of the
following markers: [0130] (i) a switch between RAF isoforms, [0131]
(ii) upregulation of RTK or NRAS signaling, [0132] (iii)
reactivation of mitogen activated protein kinase (MAPK) signaling,
[0133] (iv) the presence of a MEK activating mutation, [0134] (v)
amplification of mutant BRAF, [0135] (vi) STAT3 upregulation,
[0136] (vii) mutations in the allosteric pocket of MEK that
directly block binding of inhibitors to MEK or lead to constitutive
MEK activity, wherein the presence of one or more of the markers
confirms that the subject's cancer is refractory or resistant to
BRAF and/or MEK inhibitor therapy and that the subject would
benefit from therapy with an ERK inhibitor, which is BVD-523 or a
pharmaceutically acceptable salt thereof.
[0137] Suitable and preferred subjects are as disclosed herein. In
this embodiment, the methods may be used to identify a subject
having cancers disclosed above, including those cancers with the
mutational backgrounds, resistance profiles, and MAPK activity
identified above. Methods of identifying such mutations are also as
set forth above.
[0138] In one aspect of this embodiment, the method further
comprises administering BVD-523 or a pharmaceutically acceptable
salt thereof to a subject having one or more of the markers.
Preferably, the method additionally comprises administering to the
subject having one or more of the markers at least one additional
therapeutic agent, preferably an inhibitor of the PI3K/Akt pathway,
as disclosed herein.
[0139] An additional embodiment of the present invention is a
pharmaceutical composition for treating or ameliorating the effects
of a cancer in a subject, which cancer is refractory or resistant
to non-ERK MAPK pathway therapy. The composition comprises a
pharmaceutically acceptable carrier or diluent and an effective
amount of BVD-523 or a pharmaceutically acceptable salt
thereof.
[0140] Suitable and preferred subjects and types of non-ERK MAPK
pathway inhibitor therapy are as disclosed herein. In this
embodiment, the pharmaceutical composition may be used to treat the
cancers disclosed above, including those cancers with the
mutational backgrounds, resistance profiles, and MAPK activity
identified above. Methods of identifying such mutations are also as
set forth above.
[0141] In one aspect of this embodiment, the pharmaceutical
composition further comprises at least one additional therapeutic
agent, preferably an inhibitor of the PI3K/Akt pathway, as
disclosed herein.
[0142] Another embodiment of the present invention is a kit for
treating or ameliorating the effects of a cancer in a subject,
which cancer is refractory or resistant to non-ERK MAPK pathway
therapy. This kit comprises any pharmaceutical composition
according to the present invention packaged together with
instructions for its use.
[0143] The kits may also include suitable storage containers, e.g.,
ampules, vials, tubes, etc., for each pharmaceutical composition
and other reagents, e.g., buffers, balanced salt solutions, etc.,
for use in administering the pharmaceutical compositions to
subjects. The pharmaceutical compositions and other reagents may be
present in the kits in any convenient form, such as, e.g., in a
solution or in a powder form. The kits may further include a
packaging container, optionally having one or more partitions for
housing the pharmaceutical composition and other optional
reagents.
[0144] Suitable and preferred subjects and types of non-ERK MAPK
pathway inhibitor therapy are as disclosed herein. In this
embodiment, the kit may be used to treat the cancers disclosed
above, including those cancers with the mutational backgrounds,
resistance profiles, and MAPK activity identified herein. Methods
of identifying such mutations are as set forth above.
[0145] In one aspect of this embodiment, the kit further comprises
at least one additional therapeutic agent, preferably an inhibitor
of the PI3K/Akt pathway, as disclosed herein.
[0146] Another embodiment of the present invention is a method for
inhibiting phosphorylation of RSK in a cancer cell that is
refractory or resistant to a non-ERK MAPK pathway inhibitor. The
method comprises contacting the cancer cell with an effective
amount of BVD-523 or a pharmaceutically acceptable salt thereof for
a period of time sufficient for phosphorylation of RSK in the
cancer cell to be inhibited. In this embodiment, "contacting" means
bringing BVD-523 or a pharmaceutically acceptable salt thereof and
optionally one or more additional therapeutic agents into close
proximity to the cancer cells. This may be accomplished using
conventional techniques of drug delivery to mammals, or in the in
vitro situation by, e.g., providing BVD-523 or a pharmaceutically
acceptable salt thereof and optionally other therapeutic agents to
a culture media in which the cancer cells are located. In the ex
vivo situation, contacting may be carried out by, e.g., providing
BVD-523 or a pharmaceutically acceptable salt thereof and
optionally other therapeutic agents to a cancerous tissue.
[0147] Suitable and preferred types of non-ERK MAPK pathway
inhibitors are as disclosed herein. In this embodiment, effecting
cancer cell death may be accomplished in cancer cells having
various mutational backgrounds, resistance profiles, and MAPK
activity as disclosed above. Methods of identifying such mutations
are also as set forth above.
[0148] The methods of this embodiment, which may be carried out in
vitro, ex vivo, or in vivo, may be used to effect cancer cell
death, by e.g., killing cancer cells, in cells of the types of
cancer disclosed herein.
[0149] In one aspect of this embodiment, greater than 50% of RSK
phosphorylation is inhibited. In another aspect of this embodiment,
greater than 75% of RSK phosphorylation is inhibited. In an
additional aspect of this embodiment, greater than 90% of RSK
phosphorylation is inhibited. In a further aspect of this
embodiment, greater than 95% of RSK phosphorylation is inhibited.
In another aspect of this embodiment, greater than 99% of RSK
phosphorylation is inhibited. In an additional aspect of this
embodiment, 100% of RSK phosphorylation is inhibited.
[0150] In a further aspect of this embodiment, the cancer cell is a
mammalian cancer cell. Preferably, the mammalian cancer cell is
obtained from a mammal selected from the group consisting of
humans, primates, farm animals, and domestic animals. More
preferably, the mammalian cancer cell is a human cancer cell.
[0151] In a further aspect of this embodiment, the contacting step
comprises administering BVD-523 or a pharmaceutically acceptable
salt to a subject from whom the cancer cell was obtained.
[0152] In the present invention, an "effective amount" or a
"therapeutically effective amount" of a compound or composition
disclosed herein is an amount of such compound or composition that
is sufficient to effect beneficial or desired results as described
herein when administered to a subject. Effective dosage forms,
modes of administration, and dosage amounts may be determined
empirically, and making such determinations is within the skill of
the art. It is understood by those skilled in the art that the
dosage amount will vary with the route of administration, the rate
of excretion, the duration of the treatment, the identity of any
other drugs being administered, the age, size, and species of
mammal, e.g., human patient, and like factors well known in the
arts of medicine and veterinary medicine. In general, a suitable
dose of a compound or composition according to the invention will
be that amount of the composition, which is the lowest dose
effective to produce the desired effect. The effective dose of a
compound or composition of the present invention may be
administered as two, three, four, five, six or more sub-doses,
administered separately at appropriate intervals throughout the
day.
[0153] A suitable, non-limiting example of a dosage of a BVD-523
and other anti-cancer agents disclosed herein is from about 1 mg/kg
to about 2400 mg/kg per day, such as from about 1 mg/kg to about
1200 mg/kg per day, 75 mg/kg per day to about 300 mg/kg per day,
including from about 1 mg/kg to about 100 mg/kg per day. Other
representative dosages of such agents include about 1 mg/kg, 5
mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 35 mg/kg,
40 mg/kg, 45 mg/kg, 50 mg/kg, 60 mg/kg, 70 mg/kg, 75 mg/kg, 80
mg/kg, 90 mg/kg, 100 mg/kg, 125 mg/kg, 150 mg/kg, 175 mg/kg, 200
mg/kg, 250 mg/kg, 300 mg/kg, 400 mg/kg, 500 mg/kg, 600 mg/kg, 700
mg/kg, 800 mg/kg, 900 mg/kg, 1000 mg/kg, 1100 mg/kg, 1200 mg/kg,
1300 mg/kg, 1400 mg/kg, 1500 mg/kg, 1600 mg/kg, 1700 mg/kg, 1800
mg/kg, 1900 mg/kg, 2000 mg/kg, 2100 mg/kg, 2200 mg/kg, and 2300
mg/kg per day. The effective dose of BVD-523 and other anti-cancer
agents disclosed herein, may be administered as two, three, four,
five, six or more sub-doses, administered separately at appropriate
intervals throughout the day.
[0154] The BVD-523, other inhibitors, and various other anti-cancer
agents disclosed herein, or a pharmaceutical composition of the
present invention may be administered in any desired and effective
manner: for oral ingestion, or as an ointment or drop for local
administration to the eyes, or for parenteral or other
administration in any appropriate manner such as intraperitoneal,
subcutaneous, topical, intradermal, inhalation, intrapulmonary,
rectal, vaginal, sublingual, intramuscular, intravenous,
intraarterial, intrathecal, or intralymphatic. Further, BVD-523,
other inhibitors, and various other anti-cancer agents disclosed
herein, or a pharmaceutical composition of the present invention
may be administered in conjunction with other treatments. BVD-523,
other inhibitors, and various other anti-cancer agents disclosed
herein, or a pharmaceutical composition of the present invention
may be encapsulated or otherwise protected against gastric or other
secretions, if desired.
[0155] The pharmaceutical compositions of the invention comprise
one or more active ingredients in admixture with one or more
pharmaceutically-acceptable diluents or carriers and, optionally,
one or more other compounds, drugs, ingredients and/or materials.
Regardless of the route of administration selected, the
agents/compounds of the present invention are formulated into
pharmaceutically-acceptable dosage forms by conventional methods
known to those of skill in the art. See, e.g., Remington, The
Science and Practice of Pharmacy (21.sup.st Edition, Lippincott
Williams and Wilkins, Philadelphia, Pa.).
[0156] Pharmaceutically acceptable diluents or carriers are well
known in the art (see, e.g., Remington, The Science and Practice of
Pharmacy (21.sup.st Edition, Lippincott Williams and Wilkins,
Philadelphia, Pa.) and The National Formulary (American
Pharmaceutical Association, Washington, D.C.)) and include sugars
(e.g., lactose, sucrose, mannitol, and sorbitol), starches,
cellulose preparations, calcium phosphates (e.g., dicalcium
phosphate, tricalcium phosphate and calcium hydrogen phosphate),
sodium citrate, water, aqueous solutions (e.g., saline, sodium
chloride injection, Ringer's injection, dextrose injection,
dextrose and sodium chloride injection, lactated Ringer's
injection), alcohols (e.g., ethyl alcohol, propyl alcohol, and
benzyl alcohol), polyols (e.g., glycerol, propylene glycol, and
polyethylene glycol), organic esters (e.g., ethyl oleate and
tryglycerides), biodegradable polymers (e.g.,
polylactide-polyglycolide, poly(orthoesters), and
poly(anhydrides)), elastomeric matrices, liposomes, microspheres,
oils (e.g., corn, germ, olive, castor, sesame, cottonseed, and
groundnut), cocoa butter, waxes (e.g., suppository waxes),
paraffins, silicones, talc, silicylate, etc. Each pharmaceutically
acceptable diluent or carrier used in a pharmaceutical composition
of the invention must be "acceptable" in the sense of being
compatible with the other ingredients of the formulation and not
injurious to the subject. Diluents or carriers suitable for a
selected dosage form and intended route of administration are well
known in the art, and acceptable diluents or carriers for a chosen
dosage form and method of administration can be determined using
ordinary skill in the art.
[0157] The pharmaceutical compositions of the invention may,
optionally, contain additional ingredients and/or materials
commonly used in pharmaceutical compositions. These ingredients and
materials are well known in the art and include (1) fillers or
extenders, such as starches, lactose, sucrose, glucose, mannitol,
and silicic acid; (2) binders, such as carboxymethylcellulose,
alginates, gelatin, polyvinyl pyrrolidone, hydroxypropylmethyl
cellulose, sucrose and acacia; (3) humectants, such as glycerol;
(4) disintegrating agents, such as agar-agar, calcium carbonate,
potato or tapioca starch, alginic acid, certain silicates, sodium
starch glycolate, cross-linked sodium carboxymethyl cellulose and
sodium carbonate; (5) solution retarding agents, such as paraffin;
(6) absorption accelerators, such as quaternary ammonium compounds;
(7) wetting agents, such as cetyl alcohol and glycerol
monostearate; (8) absorbents, such as kaolin and bentonite clay;
(9) lubricants, such as talc, calcium stearate, magnesium stearate,
solid polyethylene glycols, and sodium lauryl sulfate; (10)
suspending agents, such as ethoxylated isostearyl alcohols,
polyoxyethylene sorbitol and sorbitan esters, microcrystalline
cellulose, aluminum metahydroxide, bentonite, agar-agar and
tragacanth; (11) buffering agents; (12) excipients, such as
lactose, milk sugars, polyethylene glycols, animal and vegetable
fats, oils, waxes, paraffins, cocoa butter, starches, tragacanth,
cellulose derivatives, polyethylene glycol, silicones, bentonites,
silicic acid, talc, salicylate, zinc oxide, aluminum hydroxide,
calcium silicates, and polyamide powder; (13) inert diluents, such
as water or other solvents; (14) preservatives; (15) surface-active
agents; (16) dispersing agents; (17) control-release or
absorption-delaying agents, such as hydroxypropylmethyl cellulose,
other polymer matrices, biodegradable polymers, liposomes,
microspheres, aluminum monostearate, gelatin, and waxes; (18)
opacifying agents; (19) adjuvants; (20) wetting agents; (21)
emulsifying and suspending agents; (22), solubilizing agents and
emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, oils (in particular,
cottonseed, groundnut, corn, germ, olive, castor and sesame oils),
glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty
acid esters of sorbitan; (23) propellants, such as
chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons,
such as butane and propane; (24) antioxidants; (25) agents which
render the formulation isotonic with the blood of the intended
recipient, such as sugars and sodium chloride; (26) thickening
agents; (27) coating materials, such as lecithin; and (28)
sweetening, flavoring, coloring, perfuming and preservative agents.
Each such ingredient or material must be "acceptable" in the sense
of being compatible with the other ingredients of the formulation
and not injurious to the subject. Ingredients and materials
suitable for a selected dosage form and intended route of
administration are well known in the art, and acceptable
ingredients and materials for a chosen dosage form and method of
administration may be determined using ordinary skill in the
art.
[0158] The pharmaceutical compositions of the present invention
suitable for oral administration may be in the form of capsules,
cachets, pills, tablets, powders, granules, a solution or a
suspension in an aqueous or non-aqueous liquid, an oil-in-water or
water-in-oil liquid emulsion, an elixir or syrup, a pastille, a
bolus, an electuary or a paste. These formulations may be prepared
by methods known in the art, e.g., by means of conventional
pan-coating, mixing, granulation or lyophilization processes.
[0159] Solid dosage forms for oral administration (capsules,
tablets, pills, dragees, powders, granules and the like) may be
prepared, e.g., by mixing the active ingredient(s) with one or more
pharmaceutically-acceptable diluents or carriers and, optionally,
one or more fillers, extenders, binders, humectants, disintegrating
agents, solution retarding agents, absorption accelerators, wetting
agents, absorbents, lubricants, and/or coloring agents. Solid
compositions of a similar type may be employed as fillers in soft
and hard-filled gelatin capsules using a suitable excipient. A
tablet may be made by compression or molding, optionally with one
or more accessory ingredients. Compressed tablets may be prepared
using a suitable binder, lubricant, inert diluent, preservative,
disintegrant, surface-active or dispersing agent. Molded tablets
may be made by molding in a suitable machine. The tablets, and
other solid dosage forms, such as dragees, capsules, pills and
granules, may optionally be scored or prepared with coatings and
shells, such as enteric coatings and other coatings well known in
the pharmaceutical-formulating art. They may also be formulated so
as to provide slow or controlled release of the active ingredient
therein. They may be sterilized by, for example, filtration through
a bacteria-retaining filter. These compositions may also optionally
contain opacifying agents and may be of a composition such that
they release the active ingredient only, or preferentially, in a
certain portion of the gastrointestinal tract, optionally, in a
delayed manner. The active ingredient can also be in
microencapsulated form.
[0160] Liquid dosage forms for oral administration include
pharmaceutically-acceptable emulsions, microemulsions, solutions,
suspensions, syrups and elixirs. The liquid dosage forms may
contain suitable inert diluents commonly used in the art. Besides
inert diluents, the oral compositions may also include adjuvants,
such as wetting agents, emulsifying and suspending agents,
sweetening, flavoring, coloring, perfuming and preservative agents.
Suspensions may contain suspending agents.
[0161] The pharmaceutical compositions of the present invention for
rectal or vaginal administration may be presented as a suppository,
which may be prepared by mixing one or more active ingredient(s)
with one or more suitable nonirritating diluents or carriers which
are solid at room temperature, but liquid at body temperature and,
therefore, will melt in the rectum or vaginal cavity and release
the active compound. The pharmaceutical compositions of the present
invention which are suitable for vaginal administration also
include pessaries, tampons, creams, gels, pastes, foams or spray
formulations containing such pharmaceutically-acceptable diluents
or carriers as are known in the art to be appropriate.
[0162] Dosage forms for the topical or transdermal administration
include powders, sprays, ointments, pastes, creams, lotions, gels,
solutions, patches, drops and inhalants. The active
agent(s)/compound(s) may be mixed under sterile conditions with a
suitable pharmaceutically-acceptable diluent or carrier. The
ointments, pastes, creams and gels may contain excipients. Powders
and sprays may contain excipients and propellants.
[0163] The pharmaceutical compositions of the present invention
suitable for parenteral administrations may comprise one or more
agent(s)/compound(s) in combination with one or more
pharmaceutically-acceptable sterile isotonic aqueous or non-aqueous
solutions, dispersions, suspensions or emulsions, or sterile
powders which may be reconstituted into sterile injectable
solutions or dispersions just prior to use, which may contain
suitable antioxidants, buffers, solutes which render the
formulation isotonic with the blood of the intended recipient, or
suspending or thickening agents. Proper fluidity can be maintained,
for example, by the use of coating materials, by the maintenance of
the required particle size in the case of dispersions, and by the
use of surfactants. These pharmaceutical compositions may also
contain suitable adjuvants, such as wetting agents, emulsifying
agents and dispersing agents. It may also be desirable to include
isotonic agents. In addition, prolonged absorption of the
injectable pharmaceutical form may be brought about by the
inclusion of agents which delay absorption.
[0164] In some cases, in order to prolong the effect of a drug
(e.g., pharmaceutical formulation), it is desirable to slow its
absorption from subcutaneous or intramuscular injection. This may
be accomplished by the use of a liquid suspension of crystalline or
amorphous material having poor water solubility.
[0165] The rate of absorption of the active agent/drug then depends
upon its rate of dissolution which, in turn, may depend upon
crystal size and crystalline form. Alternatively, delayed
absorption of a parenterally-administered agent/drug may be
accomplished by dissolving or suspending the active agent/drug in
an oil vehicle. Injectable depot forms may be made by forming
microencapsule matrices of the active ingredient in biodegradable
polymers. Depending on the ratio of the active ingredient to
polymer, and the nature of the particular polymer employed, the
rate of active ingredient release can be controlled. Depot
injectable formulations are also prepared by entrapping the drug in
liposomes or microemulsions which are compatible with body tissue.
The injectable materials can be sterilized for example, by
filtration through a bacterial-retaining filter.
[0166] The formulations may be presented in unit-dose or multi-dose
sealed containers, for example, ampules and vials, and may be
stored in a lyophilized condition requiring only the addition of
the sterile liquid diluent or carrier, for example water for
injection, immediately prior to use. Extemporaneous injection
solutions and suspensions may be prepared from sterile powders,
granules and tablets of the type described above.
[0167] The present invention provides treatment of cancer which is
refractory or resistant to non-ERK MAPK pathway inhibitor therapy
and discloses combinations shown to enhance the effects of ERK
inhibitors. Herein, applicants have also shown that the combination
of different ERK inhibitors is likewise synergistic. Therefore, it
is contemplated that the effects of the combinations described
herein can be further improved by the use of one or more additional
ERK inhibitors. Accordingly, some embodiments of the present
invention include one or more additional ERK inhibitors.
[0168] The present invention also provides a method of treating a
subject having an unresectable or metastatic BRAF600
mutation-positive melanoma comprising administering to the subject
600 mg BID of BVD-523 or a pharmaceutically acceptable salt
thereof.
[0169] In some embodiments of the invention, the mutation is a
BRAF.sup.V600E mutation.
[0170] The present invention also provides a composition for
treating a subject having an unresectable or metastatic BRAF600
mutation-positive melanoma, the composition comprising 600 mg of
BVD-523 or a pharmaceutically acceptable salt thereof and
optionally a pharmaceutically acceptable carrier, adjuvant, or
vehicle.
[0171] The following examples are provided to further illustrate
the methods of the present invention. These examples are
illustrative only and are not intended to limit the scope of the
invention in any way.
EXAMPLES
Example 1
Materials and Methods
[0172] Cancer cell lines were maintained in cell culture under
standard media and serum conditions. For dose escalation studies,
A375 cells were split, grown to about 40-60% confluence, and then
treated with the initial dose of the specified drug. Table 4 shows
a summary of drug treatments that were escalated.
TABLE-US-00004 TABLE 4 Summary of Treatments Being Escalated
Treatment Inhibitor 1 Trametinib (MEKi) 2 Dabrafenib (BRAFi) 3
BVD-523 (ERKi) 4 Dabrafenib (BRAFi) + Trametinib (MEKi) 5
Dabrafenib (BRAFi) + BVD-523 (ERKi) 6 Trametinib (MEKi) + BVD-523
(ERKi)
[0173] Single agent dose escalations were performed based on Little
et al., 2011 and are outlined in FIG. 20. Cells were then allowed
to grow until 70-90% confluence and split. Split ratios were kept
as "normal" as possible and reasonably consistent between
treatments (e.g. a minimum of 50% of the normal split ratio of the
parentals). Medium was refreshed every 3-4 days. When cells again
reached about 40-60% confluence, the dose was escalated. In the
event that the 40-60% window was missed, the cells were split again
and dosed once they reached 40-60% confluence. Again, medium was
refreshed every 3-4 days. The process was repeated as required
(FIG. 20).
[0174] For single agent treatments, starting concentrations and
dose increases were conducted by starting with the approximate
IC.sub.50, escalating in small increments or, gently, for the
initial 4-5 doses, doubling the dose, increasing by the same
increment for the next 4 doses, then moving to 1.5-fold increases
in concentration for subsequent doses.
[0175] For combination treatments, starting concentrations and dose
increases were conducted by starting with half of the approximate
IC.sub.50 of each compound (combination assay suggests this will
result in about 40-70% inhibition range), escalating as per single
agents (i.e. doing an initial doubling and then increasing by the
same increment for the next 4 doses, then moving to 1.5-fold
increases in concentration). Table 5 shows the projected dose
increases using these schemes.
TABLE-US-00005 TABLE 5 Projected Dose Increases--Month 1 Dab/Tram
Dab/523 Tram/523 Tram Dab BVD-523 Dab Tram Dab 523 Tram 523 Dose
(nM) (nM) (.mu.M) (nM) (nM) (nM) (.mu.M) (nM) (.mu.M) 1 1 5 0.16
2.5 0.5 2.5 0.08 0.5 0.08 2 2 10 0.32 5 1 5 0.16 1 0.16 3 3 15 0.48
7.5 1.5 7.5 0.24 1.5 0.24 4 4 20 0.64 10 2 10 0.32 2 0.32 5 5 25
0.80 12.5 2.5 12.5 0.40 2.5 0.40 6 8 38 1.2 19 4 19 0.6 4 0.6 7 11
56 1.8 28 6 28 0.9 6 0.9 8 17 84 2.7 42 8 42 1.4 8 1.4 9 25 127 4.1
63 13 63 2.0 13 2.0 10 38 190 6.1 95 19 95 3.0 19 3.0 11 57 285 9.1
142 28 142 4.6 28 4.6 12 85 427 13.7 214 43 214 6.8 43 6.8 13 128
641 20.5 320 64 320 10.3 64 10.3 14 192 961 30.8 481 96 481 15.4 96
15.4 15 288 1442 46.1 721 144 721 23.1 144 23.1 16 432 2162 69.2
1081 216 1081 34.6 216 34.6 17 649 3244 103.8 1622 324 1622 51.9
324 51.9 18 973 4865 155.7 2433 487 2433 77.8 487 77.8 19 1460 7298
233.5 3649 730 3649 116.8 730 116.8 20 2189 10947 350.3 5474 1095
5474 175.2 1095 175.2
[0176] Clonal resistant cell populations were derived from
resistant cell pools by limiting dilution.
[0177] Proliferation assays were used to track changes in
sensitivity to the escalated agent(s) at appropriate time intervals
(e.g. each month, although the timing is dependent on adequate cell
numbers being available). For proliferation assays, cells were
seeded in 96-well plates at 3000 cells per well in drug-free DMEM
medium containing 10% FBS and allowed to adhere overnight prior to
addition of compound or vehicle control. Compounds were prepared
from DMSO stocks to give a final concentration range as shown in
FIG. 2A-FIG. 2H. The final DMSO concentration was constant at 0.1%.
Test compounds were incubated with the cells for 96 hours at
37.degree. C. and 5% CO.sub.2 in a humidified atmosphere. Alamar
Blue 10% (v/v) was then added and incubated for 4 hours and
fluorescent product was detected using a BMG FLUOstar plate reader.
The average media only background value was deducted and the data
analyzed using a 4-parameter logistic equation in GraphPad Prism.
Paclitaxel was used as a positive control.
[0178] Proliferation assays for month 1 were initiated at day 28
using cells growing in the concentrations of each agent indicated
in Table 6.
TABLE-US-00006 TABLE 6 Initial Concentrations of Drugs Used in
Proliferation Assays - Month 1 Line Dab Tram BVD-523 Parental -- --
-- Tram -- 2 nM -- Dab 15 nM -- -- BVD-523 -- -- 0.48 .mu.M Tram +
Dab .sup. 5 nM 1 nM -- Dab + BVD-523 7.5 nM -- 0.24 .mu.M Tram +
BVD-523 -- 1 nM 0.16 .mu.M
[0179] Proliferation assays for month 2 were initiated at day 56
using cells growing in the concentrations of each agent indicated
in Table 7.
TABLE-US-00007 TABLE 7 Initial Concentrations of Drugs Used in
Proliferation Assays - Month 2 Line Dab Tram BVD-523 Parental -- --
-- Tram -- 8 nM -- Dab 127 nM -- -- BVD-523 -- -- 0.8 .mu.M Tram +
Dab 10 nM 2 nM -- Dab + BVD-523 12.5 nM -- 0.4 .mu.M Tram + BVD-523
-- 2 nM 0.32 .mu.M
[0180] At the end of the 3 month escalation period, cultures were
maintained at the top concentration for 2 weeks prior to the final
round of proliferation assays and potential single cell cloning. As
the proliferation assays/single cell cloning required actively
proliferating cells, for treatments where cells were proliferating
very slowly at the top concentration or that were only recently
escalated, a backup culture was also maintained at a lower
concentration (Table 8). For the BVD-523 treatment, where cells
appeared to have almost completely stopped growing and looked
particularly fragile at the top concentration (1.8 .mu.M), cultures
were maintained at a lower concentration for the 2 week period.
TABLE-US-00008 TABLE 8 Details of Treatments Being Cultured at a
Fixed Concentration for 2 Weeks Treatment Inhibitor Culture 1
Backup Culture 1 Tram 160 nM 80 nM 2 Dab 3.2 .mu.M -- 3 BVD-523 1.2
.mu.M 0.8 .mu.M 4 Dab + Tram D: 160 nM D: 80 nM T: 30 nM T: 16 nM 5
Dab + BVD-523 D: 42 nM D: 28 nM 523: 1.4 .mu.M 523: 0.9 .mu.M 6
Tram + BVD-523 T: 4 nM T: 2.5 nM 523: 0.6 .mu.M 523: 0.4 .mu.M
[0181] Proliferation assays for month 3 used cells growing in the
concentrations of each agent indicated in Table 9.
TABLE-US-00009 TABLE 9 Initial Concentrations of Drugs Used in
Proliferation Assays - Month 3 Line Dab Tram BVD-523 Parental -- --
-- Tram -- 160 nM -- Dab 3.2 .mu.M -- -- BVD-523 -- -- 1.2 .mu.M
Tram + Dab 80 nM 16 nM -- Dab + BVD-523 28 nM -- 0.9 .mu.M Tram +
BVD-523 -- 2.5 nM 0.4 .mu.M
[0182] For combination studies, A375 cells (ATCC) were seeded into
triplicate 96-well plates at a cell density of 3000 cells/well in
DMEM plus 10% FBS and allowed to adhere overnight prior to addition
of test compound or vehicle control. Combinations were tested using
a 10.times.8 dose matrix with a final DMSO concentration of 0.2%. A
96 hour assay incubation period followed, with subsequent addition
of Alamar Blue 10% (v/v) and 4 hours incubation prior to reading on
a fluorescent plate reader. After reading Alamar Blue, the
medium/Alamar Blue mix was flicked off and 100 .mu.l of
CellTiter-Glo/PBS (1:1) added and the plates processed as per the
manufacturers instructions (Promega). Media only background values
were subtracted before the data was analysed. The Bliss additivity
model was then applied.
[0183] In brief, predicted fractional inhibition values for
combined inhibition were calculated using the equation
C.sub.bliss=A+B-(A.times.B) where A and B are the fractional
inhibitions obtained by drug A alone or drug B alone at specific
concentrations. C.sub.bliss is the fractional inhibition that would
be expected if the combination of the two drugs were exactly
additive. C.sub.bliss values are subtracted from the experimentally
observed fractional inhibition values to give an `excess over
Bliss` value. Excess over Bliss values greater than 0 indicate
synergy, whereas values less than 0 indicate antagonism. Excess
over Bliss values are plotted as heat maps.+-.SD.
[0184] The single and combination data are also presented as
dose-response curves generated in GraphPad Prism (plotted using %
viability relative to DMSO only treated controls).
[0185] For focused combination studies, the Alamar Blue viability
assays were performed as described above for combination studies.
Additionally, Caspase-Glo 3/7 assays were performed. In brief,
HCT116 cells were seeded in triplicate in white 96-well plates at a
cell density of 5000 cells/well in McCoy's 5A plus 10% FBS. A375
cells were seeded at a density of 5000 cells/well in DMEM plus 10%
FBS. Cells were allowed to adhere overnight prior to addition of
test compound or vehicle control. The final concentration of DMSO
was 0.2%, and 800 nM staurosporine was included as a positive
control. 24 and 48 hour assay incubation periods were used. Then,
Caspase-Glo.RTM. 3/7 50% (v/v) was added, plates were mixed for 5
minutes on an orbital shaker and incubated for 1 hour at room
temperature prior to reading on a luminescent plate reader. Media
only background values were subtracted before the data was
analysed.
[0186] For Differential Scanning Fluorimetry, SYPRO orange
(5,000.times. solution, Invitrogen) was diluted (1:1,000) in buffer
solution (10 mM HEPES, 150 mM NaCl, pH 7.5). HisX6 tagged proteins
included inactive ERK2, active ERK2 (ppERK2), or p38a at a final
concentration of 1 .mu.M. The protein/dye solution and compounds in
100% DMSO were added to wells (2% v/v final DMSO concentration) to
achieve the desired final concentrations, mixed, and placed into an
RT-PCR instrument. Next, a melting curve was run from 25-95.degree.
C. at a rate of 1.degree. C. per minute and the melting temperature
(Tm) was determined for each protein in the absence or presence of
compounds. The change in Tm (.DELTA.Tm) in the presence of various
drug concentrations is presented.
[0187] For Ki determination of ERK1, activated ERK1 (10 nM) was
incubated with various concentrations of the compounds in 2.5%
(v/v) DMSO for 10 minutes at 30.degree. C. in 0.1 M HEPES buffer
(pH 7.5), 10 mM MgCl.sub.2, 2.5 mM phosphoenolpyruvate, 200 .mu.M
nicotinamide adenine dinucleotide (NADH), 150 .mu.g/mL pyruvate
kinase, 50 .mu.g/mL lactate dehydrogenase, and 200 .mu.M Erktide
peptide. The reaction was initiated by the addition of 65 .mu.M of
ATP. Decreased absorbance rate (340 nm) was monitored and the
IC.sub.50 was determined as a function of inhibitor
concentration.
[0188] For Ki determination of ERK2, the inhibitory activity of
BVD-523 against ERK2 was determined using a radiometric assay, with
final concentration of the components being 100 mM HEPES (pH 7.5),
10 mM MgCl.sub.2, 1 mM dithiothreitol (DTT), 0.12 nM ERK2, 10 .mu.M
myelin basic protein (MBP), and 50 .mu.M .sup.33P-.gamma.-ATP. All
reaction components, with the exception of ATP and MBP, were
premixed and aliquoted (33 .mu.L) into a 96-well plate. A stock
solution of compound in DMSO was used to make up to 500-fold
dilutions; a 1.5-.mu.L aliquot of DMSO or inhibitor in DMSO was
added to each well. The reaction was initiated by adding the
substrates .sup.33P-.gamma.-ATP and MBP (33 .mu.L). After 20
minutes the reaction was quenched with 20% (w/v) tricholoracetic
acid (TCA) (55 .mu.L) containing 4 mM ATP, transferred to the GF/B
filter plates, and washed 3 times with 5% (w/v) TCA). Following the
addition of Ultimate Gold.TM. scintillant (50 .mu.L), the samples
were counted in a Packard TopCount. From the activity versus
concentration titration curve, the Ki value was determined by
fitting the data to an equation for competitive tight binding
inhibition kinetics using Prism software, version 3.0.
[0189] For IC.sub.50 determination of ERK2, activity was assayed by
a standard coupled-enzyme assay. The final concentrations were as
follows: 0.1 M HEPES (pH 7.5), 10 mM MgCl.sub.2, 1 mM DTT, 2.5 mM
phosphoenolpyruvate, 200 .mu.M NADH, 50 .mu.g/mL pyruvate kinase,
10 .mu.g/mL lactate dehydrogenase, 65 .mu.M ATP, and 800 .mu.M
peptide (ATGPLSPGPFGRR). All of the reaction components except ATP
were premixed with ERK and aliquoted into assay-plate wells.
BVD-523 in DMSO was introduced into each well, keeping the
concentration of DMSO per well constant. BVD-523 concentrations
spanned a 500-fold range for each titration. The assay-plate was
incubated at 30.degree. C. for 10 minutes in the plate reader
compartment of the spectrophotometer (molecular devices) before
initiating the reaction by adding ATP. The absorbance change at 340
nm was monitored as a function of time; the initial slope
corresponds to the rate of the reaction. The rate versus
concentration of the BVD-523 titration curve was fitted either to
an equation for competitive tight-binding inhibition kinetics to
determine a value for Ki or to a 3-parameter fit to determine the
IC.sub.50 using Prism software, version 3.0.
[0190] For apoptosis assays, cells were plated at 2.times.10.sup.4
cells per well in a 96-well plate and allowed to attach overnight
or grow to 50% confluency. Cells were treated with a serial
dilution of BVD-523 in media (final volume 200 .mu.L, concentration
ranges 4-0.25 .mu.M) and incubated for 48 hours in a 37.degree. C.
CO.sub.2 incubator. Cells were washed with 100 .mu.L of PBS, and 60
.mu.L of radioimmunoprecipitation assay buffer was added (50 mM
Tris-HCl, pH 8.0, 150 mM NaCl, 1.0% [w/v] NP-40, 0.5% [w/v] sodium
deoxycholate, 1% [w/v] SDS), then incubated for 10 minutes at
4.degree. C. to lyse the cells. A 30-.mu.L lysate aliquot was added
to 100 .mu.L of caspase assay buffer (120 mM HEPES, 12 mM EDTA, 20
mM dithiothreitol, 12.5 .mu.g/mL AC-DEVD-AMC caspase substrate) and
incubated at RT from 4 hours to overnight. The plate was read in a
fluorimeter (excitation wavelength 360 nm, emission wavelength 460
mm). The remaining 30 .mu.L of lysate was analyzed for total
protein content using the BioRad Protein Assay Kit
(sample-to-working reagent ratio of 1:8). Final normalized caspase
activity was derived as fluorescence units per .mu.g protein and
converted to a fold increase in caspase activity when compared with
DMSO controls.
[0191] For measurement of antitumor activity in A375 xenografts,
xenografts were initiated with A375 cells maintained by serial
subcutaneous transplantation in female athymic nude mice. Each test
mouse received an A375 tumor fragment (1 mm.sup.3) implanted
subcutaneously in the right flank. Once tumors reached target size
(80-120 mm.sup.3), animals were randomized into treatment and
control groups, and drug treatment was initiated.
[0192] To evaluate BVD-523 monotherapy, BVD-523 in 1% (w/v)
carboxymethylcellulose (CMC) was administered orally, per os
(p.o.), BID at doses of 5, 25, 50, 100, or 150 mg/kg. Oral
temozolomide was administered as a positive reference compound at
75 or 175 mg/kg once daily (QD) for a total of five treatments
(QD.times.5).
[0193] The efficacy of BVD-523 in combination with dabrafenib was
evaluated in mice randomized into 9 groups of 15 and 1 group of 10
(Group 10). Dabrafenib was administered p.o. at 50 or 100 mg/kg QD
and BVD-523 was administered p.o. at 50 or 100 mg/kg BID, alone and
in combination, until study end; vehicle-treated and
temozolomide-treated (150 mg/kg QD.times.5) control groups were
also included. Combination dosing was stopped on Day 20 to monitor
for tumor regrowth. Animals were monitored individually and
euthanized when each tumor reached an endpoint volume of 2000
mm.sup.3, or the final day (Day 45), whichever came first, and
median time to endpoint (TTE) calculated. The combination was also
evaluated in an upstaged A375 model where larger tumors in the
range 228-1008 mm.sup.3 were evaluated. Here, mice were randomized
into 1 group (Group 1) of 14 and 4 groups (Groups 2-5) of 20.
Dosing was initiated on Day 1 with dabrafenib plus BVD-523 (25
mg/kg dabrafenib+50 mg/kg BVD-523 or 50 mg/kg dabrafenib+100 mg/kg
BVD-523), with each agent given p.o. BID until study end. The study
included 50-mg/kg dabrafenib and 100-mg/kg BVD-523 monotherapy
groups as well as a vehicle-treated control group. Tumors were
measured twice weekly. Combination dosing was stopped on Day 42 to
monitor for tumor regrowth through study end (Day 60). Treatment
outcome was determined from % TGD, defined as the percent increase
in median TTE for treated versus control mice, with differences
between groups analyzed via log rank survival analysis. For TGI
analysis, % TGI values were calculated and reported for each
treatment (T) group versus the control (C) using the initial (i)
and final (f) tumor measurements based on the following formula: %
TGI=1-Tf-Ti/Cf-C. Mice were also monitored for CR and PR responses.
Animals with a CR at the end of the study were additionally
classified as TFS.
[0194] For measurement of BVD-523 activity in Colo205 xenografts,
human Colo205 cells were cultured in RPMI 1640 supplemented with
10% (v/v) fetal bovine serum (FBS), 100 units/mL penicillin, 100
.mu.g/mL streptomycin (Invitrogen), and 2 mM L-glutamine. Cells
were cultured for fewer than four passages prior to implantation.
Female athymic nude mice (19-23 g) were injected subcutaneously
with 2.times.10.sup.6 Colo205 cells into the right dorsal axillary
region on Day 0.
[0195] Mice with an approximate tumor volume of 200 mm.sup.3 were
randomized into 6 experimental groups. Vehicle control, 1% CMC
(w/v), was prepared weekly. BVD-523 was suspended in 1% (w/v) CMC
at the desired concentration and homogenized on ice at 6,500 rpm
for 50 minutes. BVD-523 suspensions were prepared weekly and
administered p.o. BID at total daily doses of 50, 100, 150, and 200
mg/kg (n=12/group) on an 8- or 16-hour dosing schedule for 13 days.
The vehicle control (n=12) was administered using the same dosing
regimen. CPT-11 was administered as a positive reference compound
(n=12). Each 1 mL of CPT-11 injection contained 20 mg irinotecan,
45 mg sorbitol, and 0.9 mg lactic acid. CPT-11 was administered at
100 mg/kg/day intraperitoneally every 4 days for 2 consecutive
doses.
[0196] For measurement of ERK1/2 Isotope-Tagged Internal Standard
(ITIS) Mass Spectrometry in Colo205 Xenografts, frozen tumors were
lysed in 10 volumes of ice cold lysis buffer (10 mM TRIS-HCl, pH
8.0, 10 mM MgCl.sub.2, 1% (v/v) Triton X-100, Complete.TM. Protease
Inhibitor Cocktail [Roche, cat. No. 1836170], Phosphatase Inhibitor
Cocktail I [Sigma, cat. No. P-2850], Phosphatase Inhibitor Cocktail
II [Sigma cat. No. 5726], and benzonase [Novagen cat. No. 70664]).
Lysates were clarified by centrifugation (100,000.times.g for 60
minutes at 4.degree. C.) and the supernatants adjusted to 2 mg/mL
with lysis buffer. ERK1 was immunoprecipitated using
agarose-coupled and pan-anti-ERK1 (Santa Cruz Biotechnology cat.
No. sc-93ac) antibodies. Immunoprecipitated proteins were resolved
by SDS-PAGE and stained with SYPRO Ruby (Invitrogen), and the ERK
bands excised via razor. Gel slices were washed in 300 .mu.L of 20
mM NH.sub.4HCO.sub.3, diced into small pieces, and placed in Page
Eraser Tip (The Nest Group cat no. SEM0007). Gel fragments were
reduced and alkylated prior to trypsin digestion. Tryptic fragments
were isolated in 75 .mu.L of 50% (v/v) Acetonitrile, 0.2% (v/v)
trifluoroacetic acid and the resulting sample concentrated to 0-10
.mu.L in a SpeedVac.
[0197] For ITIS analysis, digested samples were spiked with
heavy-atom labeled peptide standards and fractional phosphorylation
was quantified by coupled liquid chromatography-tandem mass
spectrometry (MS). Nanocapillary chromatography was performed using
a Rheos 2000 binary pump from Flux Instruments delivering nanoscale
flow after 1:750 splitting, an LC Packings Inertsil nano-precolumn
(C18, 5 mm, 100 .ANG., 30 mm ID.times.1 mm), and a New Objective
PicoFrit AQUASIL resolving column (C18, 5 mm, 75/15 mm ID.times.10
cm), which also served as an electrospray ionization (ESI) emitter.
An Applied Biosystem API 3000 mass spectrometer coupled with a
nano-ESI source was used for MS analysis. An in-house-made gas
nozzle connected to a nebulizing gas source was used to help steady
nano-flow spray. Data were acquired in a multiple reaction
monitoring (MRM) mode: nebulizing gas at 3; curtain gas at 7;
collision gas at 5; ion spray voltage at 2150 volts, exit potential
at 10 volts; Q1/Q3 resolution Low/Unit; and dwell time of 65 msec
for all MRM channels. All raw MS data were processed using a
combination of the Analyst software suite from Applied Biosystem
and custom tools.
[0198] For assessment of drug sensitivity in cell-line models of
acquired resistance, drug sensitivity of dose-escalated A375 cells
and isogenic RKO cells was assessed in 96-hour proliferation
assays. RKO isogenic cells (McCoy's 5A containing 10% [v/v] FBS) or
dose-escalated A375 cells (DMEM containing 10% FBS were seeded into
96-well plates and allowed to adhere overnight prior to addition of
compound or vehicle control. Note that the dose-escalated A375
cells were seeded in the absence of inhibitor. Compounds were
prepared from 0.1% (v/v) DMSO stocks to give a final concentration
as indicated. Test compounds were incubated with the cells for 96
hours at 37.degree. C. in a 5% CO.sub.2 humidified atmosphere. For
the RKO cells, CellTiter-Glo.RTM. reagent (Promega) was added
according to manufacturer's instructions and luminescence detected
using a BMG FLUOstar plate reader. For the A375 assays Alamar blue
(ThermoFisher) 10% (v/v) was added and incubated for 4 h, and
fluorescent product was then detected using a BMG FLUOstar. The
average media only background value was deducted and the data
analyzed using a 4-parameter logistic equation in GraphPad
Prism.
[0199] IC.sub.50 Determination of ERK1 was measured in a final
reaction volume of 25 .mu.L. ERK1 (human) (5-10 mU) was incubated
with 25 mM Tris (pH 7.5), 0.02 mM ethyleneglycoltetracetic acid,
250 .mu.M peptide, 10 mM Mg acetate, and .gamma.-.sup.33P-ATP
(specific activity approximately 500 cpm/pmol, concentration as
required). Adding Mg ATP initiated the reaction. After incubation
for 40 minutes at room temperature (RT), the reaction was stopped
by adding 5 .mu.L of a 3% (w/v) phosphoric acid solution. Then, 10
.mu.L of the reaction was spotted onto a P30 filtermat, and washed
3 times for 5 minutes in 75 mM of phosphoric acid then once in
methanol before drying and scintillation counting.
[0200] RKO MEK1 Q56P Isogenic cells were produced by Horizon
Discovery (Cambridge, UK; #HD 106-019) using a recombinant
AAV-mediated gene targeting strategy. Briefly, rAAV virus was
generated following transfection of the appropriate targeting
vector and helper vectors in HEK293T cells, purified using an AAV
purification kit (Virapur, San Diego, USA) and titrated using qPCR.
Parental homozygous RKO cells (homozygous wild type for MEK1) were
then infected with rAAV virus and clones that had integrated the
selection cassette were identified by G418 selection and expanded.
Correctly targeted clones that were heterozygous for knock-in of
the MEK1 Q56P point mutation into a single allele were identified
by PCR and sequencing.
[0201] Isogenic SW48 cell lines heterozygous for knock-in of mutant
KRAS (De Roock et al 2010, JAMA, 304, 1812-1820) were obtained from
Horizon Discovery (Catalogue numbers; HD 103-002, HD 103-006 HD
103-007, HD 103-009, HD 103-010, HD 103-011, HD 103-013). For
proliferation assay, cells were seeded into 96-well plates in
McCoy's 5A medium supplemented with 10% FBS and allowed to adhere
overnight prior to addition of compound or vehicle control. Test
compounds were incubated with the cells for 96 hours at 37.degree.
C. in a 5% CO.sub.2 atmosphere. Viability was then assessed using
Alamar blue.
[0202] The proprietary KinaseProfiler assay was conducted at
Upstate Discovery and employed radiometric detection similar to
that employed by Davies et al, was used to profile the selectivity
of BVD-523 against a panel of 70 kinases.
[0203] A drug sensitivity analysis was carried out as part of The
Genomics of Drug Sensitivity in Cancer Project using
high-throughput screening, as previously described (Yang et al.
2013).
[0204] For Western blot analysis, A375 cells were seeded onto 10 cm
dishes in Dulbecco's Modified Eagle's Medium plus 10% (v/v) FBS.
Cells were allowed to adhere overnight prior to the addition of
test compound or vehicle. For experiments with RKO cells, these
cells were seeded in 6-well plates or 10 cm dishes with McCoy's
5A+10% (v/v) FBS. Cells were then treated at the desired
concentration and duration. Cells were harvested by trypsinization,
pelleted, and snap frozen. Lysates were prepared with RIPA buffer
supplemented with protease and phosphatase inhibitor cocktails
(Roche), clarified by centrifugation at 11,000 rpm for 10 minutes,
and quantitated by bicinchoninic acid assay. Samples were resolved
by SDS-PAGE, blotted onto polyvinylidene difluoride membranes, and
probed using antibodies (i.e., pRB [Ser780], cat. no. 9307; CCND1,
cat. no. ab6152; BCL-xL, cat. no. 2762; PARP, cat. no. 9542; DUSP6,
cat. no. 3058S) directed to the indicated targets.
[0205] For Reverse Phase Protein Analysis (RPPA), A375, MIAPaCa-2,
HCT116, Colo205, HT-29, and AN3Ca cells (ATCC) were plated at 80%
confluence, allowed to recover overnight (MIAPaCa-2 cells were
plated at 30% confluence and allowed to recover for 3 days), then
treated with 10 .mu.M of each compound (i.e., BVD-523, SCH722984,
GDC-0994, or Vx-11e) for 6 hours at 37.degree. C. Control wells
were treated with DMSO at 0.1% (v/v) for 6 hours prior to cell
lysate generation. Samples were then analyzed using reverse-phase
protein microarray technology (Theranostics Health).
[0206] For analysis of pERK IHC in Colo205 xenografts, xenograft
tumors were processed overnight in 70% through 100% graded
ethanols, cleared in two changes of xylene, infiltrated with
paraffin, and embedded into paraffin blocks. Then, 5-.mu.m sections
were cut and placed onto positively charged glass slides and baked
for at least 30 minutes, but not longer than 1 hour, at 60.degree.
C. A single section from each animal and dose group was probed with
anti-phospho p42/p44 MAPK antibody (pERK [1:100], CST; Cat no.
9101; Lot no. 16), counterstained with hematoxylin, and then
analyzed microscopically using a Zeiss Axioplan 2 microscope. An
isotype control (rabbit, Zymed laboratories, catalog no. 08-6199,
lot no. 40186458) was run as a negative control.
[0207] For FACS analysis, cells were scraped and pelleted at 1,500
rpm for 5 minutes, then re-suspended in 1 mL of buffer and frozen
at -70.degree. C. The frozen cells were thawed and centrifuged
again, followed by 10 minutes of re-suspension in 0.25 mL of Buffer
A (trypsin in spermine tetrahydrochloride detergent buffer) to
disaggregate cell clumps and digest cell membranes and
cytoskeletons. Buffer B (trypsin inhibitor and Ribonuclease I in
buffer, 0.2 mL) was added for 10 minutes in the dark. The resulting
DNA-stained nuclei were filtered and analyzed by FACS. The
histograms were analyzed to establish the proportion of cells in
the G1, S, and G2/M phases of the cell cycle based on the presence
of n and 2n DNA (or higher) content.
[0208] For measurement of in vitro combination activity, five
thousand G-361 cells were seeded into triplicate 96-well plates
containing McCoy's 5A with 10% (v/v) FBS and allowed to adhere
overnight. The vemurafenib/BVD-523 combination was tested using a
10.times.8 dose matrix. Compounds were incubated with the cells for
72 hours at 37.degree. C. in a 5% CO.sub.2 humidified atmosphere.
CellTiter-Glo reagent was added according to manufacturer's
instructions and luminescence detected using a MBG FLUOstar plate
reader. The interactions across the dose matrix were determined by
the Loewe Additivity and Bliss independence models using Horizon's
Chalice Combination Analysis Software.
[0209] For generating compound resistance in vitro by dose
escalation, A375 parental cells (ATCC CRL-1619) were grown to
.about.40-60% confluence in Dulbecco's Modified Eagle's Medium
(DMEM) supplemented with 10% heat-inactivated FBS and
penicillin/streptomycin, then treated with initial doses of
BVD-523, trametinib, or dabrafenib either alone or in combination
at or slightly below each compound's IC.sub.50; for combination
studies, initial dosing was half of each compound's IC.sub.50.
Cells were allowed to grow until .about.70-90% confluence and
split; medium was refreshed every 3-4 days. When cells again
reached .about.40-60% confluence, the dose was escalated by the
same increment (equivalent to the starting concentration) then
moved to 1.5-fold increases in concentration followed by a further
move to 2-fold increases if the cells continued to adapt rapidly
(e.g., the first six doses of the dabrafenib escalation were: 5,
10, 15, 20, 25, and 37.5 nM). This process was repeated as
required.
[0210] Cell viability assays for FIG. 30A were performed by a
Resazurin (Alamar Blue) metabolic assay after 5 days in drug in
full serum under high glucose conditions. Cells were seeded in
384-well microplates at .about.15%-50% confluence in medium with
10% FBS and penicillin/streptavidin plus high glucose (18-25 mM).
The optimal cell number for each cell line was determined to
optimize growth during drugging. For adherent cell lines, after
overnight incubation cells were treated with 9 concentrations of
each compound (2-fold dilutions series) using liquid handling
robotics, and returned to the incubator for assay at a 96-h time
point. For suspension cell lines, cells were treated with compound
immediately after plating and returned to the incubator for a 96-h
time point. Cells were then stained with 55 .mu.g/ml Resazurin
(Sigma) prepared in glutathione-free media for 4 hours.
Quantitation of fluorescent signal intensity was performed using a
fluorescent plate reader at excitation and emission wavelengths of
535/595 nm for Resazurin. All screening plates were subjected to
stringent quality control measures. Effects on cell viability were
measured and a curve-fitting algorithm was applied to the raw
dataset to derive a multi-parameter description of drug response,
including the half maximal inhibitory concentration (IC.sub.50).
IC.sub.50 is expressed in natural log of the IC.sub.50 in .mu.M
(LN_IC.sub.50; EXP returns IC.sub.50 in .mu.M). Extrapolation of
the IC.sub.50 was allowed for where it yielded very high values. If
desired the data was restricted to the tested concentration range
by capping IC.sub.50 values at the maximum tested concentration
(and the minimum tested concentration for low values).
[0211] For efficacy testing of BVD-523 in a patient-derived
xenograft (AT052C) representing melanoma from a BRAF.sup.V600E
patient that had become clinically refractory to vemurafenib. Tumor
fragments were harvested from host animals and implanted into
immune-deficient mice. The study was initiated at a mean tumor
volume of approximately 170 mm.sup.3, at which point the animals
were randomized into four groups including a control (1% [v/v] CMC
p.o., BID.times.31) and three treatment groups (BVD-523 [100
mg/kg], dabrafenib [50 mg/kg], or BVD-523/dabrafenib [100/50
mg/kg], n=10/group); All treatment drugs were administered p.o. on
a BID.times.31 schedule.
[0212] For IC.sub.50 determination for the inhibition of
PMA-stimulated RSK1 phosphorylation by BVD-523 in human whole blood
samples, IC.sub.50 values for the inhibition of PMA stimulated RSK1
phosphorylation by BVD-523 were determined for 10 healthy donors
(aged 22-61 years) using an 8-point concentration curve ranging
from 10 .mu.M to 5 nM of BVD-523. Controls consisted of 3
unstimulated samples and 3 PMA-stimulated samples for each donor.
Both phosphor-RSK (pRSK) and total RSK levels were determined and
data were calculated using pRSK/RSK levels for each sample.
[0213] Thirty milliliters of blood was drawn from each donor into
sodium heparin vacutainers. One mL of whole blood was added to each
of twenty-two 2-mL microtubes per donor. The microtubes tubes were
labeled with the donor number (1 through 10) and the subsequent
treatment designation: "A" for PMA stimulation only (maximum), "B"
for BVD-523-containing samples that received PMA stimulation; and
"C" for the unstimulated samples (minimum). Dimethyl sulfoxide
(DMSO) was added to all tubes in groups A and C to a final
concentration of 0.1%. Samples were then rocked gently at room
temperature.
[0214] BVD-523 (10 mM in 100% DMSO) was serially diluted with
3-fold dilutions into 100% DMSO. These serially diluted BVD-523
samples in 100% DMSO were then diluted 10-fold in Dulbecco's
Modified Eagle Medium containing 10% fetal bovine serum and
penicillin/streptomycin/glutamine, and 10 .mu.L of each of these
working solutions was added per mL of blood for each designated
BVD-523 concentration. Each concentration of BVD-523 was run in
duplicate, two 1-mL blood samples each, yielding 16 total samples
for the full 8-point concentration curve. Samples were then rocked
gently at room temperature for a minimum of 2 hours but not longer
than 3 hours.
[0215] Human whole blood samples in groups A and B for all donors
were stimulated with PMA at a final concentration of 100 nM for 20
minutes at room temperature. Samples in group C were not treated
with PMA but were rocked and handled as all other samples.
[0216] Upon completion of PMA treatment for each sample, peripheral
blood mononuclear cells were isolated from the human whole blood.
One mL of blood from each sample was gently layered onto 0.75 mL of
room-temperature Histopaque 1077 in a 2-mL microcentrifuge tube.
The samples were centrifuged for 2 minutes at 16,000.times.g in an
Eppendorf microcentrifuge. The interface and upper layers were
removed and added to tubes containing 1 mL of cold Dulbecco's
phosphate-buffered saline (DPBS). These samples were then
centrifuged for 30 seconds at 16,000.times.g to pellet the cells.
The buffer supernatant was removed by aspiration and the pellets
were re-suspended in 1 mL of cold DPBS. The pellets from each
sample were then re-pelleted as above. The buffer was removed by
aspiration and the pellets were lysed as indicated below.
[0217] Complete lysis buffer consisted of Meso Scale Discovery Tris
lysis buffer, 1.times. Halt Protease inhibitor cocktail, 1.times.
Phosphatase inhibitor cocktail 2, 1.times. Phosphatase inhibitor
cocktail 3, 2 mM phenylmethanesulfonyl fluoride, and 0.1% sodium
dodecyl sulfate. Lysis buffer was kept on ice and made fresh for
each sample group. Final cell pellets were lysed by the addition of
120 L of complete lysis buffer. Samples were vortexed until the
cell pellet disappeared and then flash frozen on dry ice. Samples
were stored at -20.degree. C. prior to measurement of pRSK and
total RSK by ELISA.
[0218] For the pRSK ELISA (PathScan), thawed lysates were combined
1:1 with sample diluent (provided in ELISA kit): 120 .mu.L of
lysate added to 120 .mu.L of sample diluent in a round bottom
96-well plate. This combination was then transferred to the pRSK
microwells at 100 .mu.L per well. For the total RSK ELISA
(PathScan), 20 .mu.L of the lysate already diluted 1:1 in sample
diluent was further diluted in 200 .mu.L of sample diluent in a
round bottom 96-well plate. This combination was then transferred
to the total RSK microwells at 100 .mu.L per well. The plates were
sealed with a plate seal and incubated 16 to 18 hours at 4.degree.
C., a time that was shown to yield the best detection of the target
protein. Both ELISAs were developed according to the kit
instructions.
[0219] Patients aged .gtoreq.18 years were eligible for
participation if they had noncurable, histologically confirmed
metastatic or advanced stage malignant tumors; an ECOG performance
status of 0 or 1; adequate renal, hepatic, bone marrow, and cardiac
function; and a life expectancy .gtoreq.3 months. Patients may have
received up to 2 prior lines of chemotherapy for their metastatic
disease. Exclusion criteria were known uncontrolled brain
metastases; gastrointestinal conditions which could impair
absorption of study medication; history or current evidence/risk of
retinal vein occlusion or central serous retinopathy; and
concurrent therapy with drugs known to be strong inhibitors of
CYP1A2, CYP2D6, and CYP3A4 or strong inducers of CYP3A4. All
participants provided informed consent prior to initiation of any
study procedures.
[0220] Patients that received at least one dose of BVD-523 were
included in the analysis using SAS (version 9.3) software. The data
cutoff was Dec. 1, 2016. This study is registered with
ClinicalTrials.gov, number NCT01781429.
[0221] The present invention presents data from an open-label,
multicenter phase I study to assess the safety, pharmacokinetics,
and pharmacodynamics of escalating doses of BVD-523 in patients
with advanced malignancies. The dosing regimen combined both
accelerated titration and standard cohort 3+3 dose escalation
schema, which were used jointly to identify the MTD and RP2D of
BVD-523 in patients with advanced solid tumors. One to 6 patients
per treatment cohort were assigned to receive sequentially higher
oral doses of BVD-523 on a BID schedule (12-hour intervals) in
21-day cycles, starting at a dose of 10 mg BID. BVD-523 was
administered BID continuously in 21-day cycles at the following
doses: 10 mg (n=1); 20 mg (n=1); 40 mg (n=1); 75 mg (n=1); 150 mg
(n=1); 300 mg (n=4); 600 mg (n=7); 750 mg (n=4); and 900 mg
(n=7).
[0222] Patients received BID oral doses until disease progression,
unacceptable toxicity, or a clinical observation satisfying another
withdrawal criterion. Dose escalations occurred in up to 100%
increments in single-patient cohorts until 1 patient experienced a
.gtoreq.Grade 2 toxicity (excluding alopecia or diarrhea). Cohorts
were then expanded to at least 3 patients each and subsequent
dose-escalation increments were reduced from up to 100% to a
maximum of 50%. When at least 1 patient in a 3-patient cohort
experienced a DLT, up to 3 additional patients were treated at this
dose level. When more than 1 DLT occurred in .ltoreq.6 patients,
this dose level was defined as the nontolerated dose and dose
escalation was stopped. Intrapatient dose escalation was allowed,
provided the patients receiving the highest current dose had been
observed for at least 3 weeks and dose-limiting side effects were
reported in fewer than 2 of 6 patients assigned to a given dose.
Patients experiencing DLTs or unacceptable toxicity had their
treatment interrupted until the toxicity returned to .ltoreq.Grade
1. Resumption of BVD-523 treatment was then initiated at the next
lower dose level tested or at a 20% to 30% dose decrease, aligning
with capsule dosage.
[0223] The primary objective of the phase I study was to define the
safety and tolerability of BVD-523 by determining the dose-limiting
toxicities, the MTD, and the RP2D. The secondary objectives
included the determination of the pharmacokinetic profile of
BVD-523 in patients with advanced malignancies and the
investigation of any preliminary clinical effects on tumor
response, as assessed by physical or radiologic exam using RECIST
v1.1. The exploratory objectives included evaluation of
pharmacodynamic marker (biomarker) measures and investigation of
preliminary clinical effects on tumor response assessed by
.sup.18F-FDG-PET as indicated.
[0224] For determination of MTD, DLT, and RP2D, MTD was defined as
the highest dose cohort at which .ltoreq.33% of patients
experienced BVD-523-related DLTs in the first 21 days of treatment.
DLT was defined as a BVD-related toxicity in the first 21 days of
treatment that resulted in .gtoreq.Grade 4 hematologic toxicity for
>1 day; Grade 3 hematologic toxicity with complications (e.g.,
thrombocytopenia with bleeding); .gtoreq.Grade 3 nonhematologic
toxicity, except untreated nausea, vomiting, constipation, pain,
and rash (these become DLTs if the AE persisted despite adequate
treatment); or a treatment interruption exceeding 3 days in Cycle 1
(or the inability to being in Cycle 2 for >7 days) due to
BVD-523-related toxicity.
[0225] The RP2D could be as high as the MTD and was determined in
discussion with the clinical investigators, the medical monitor,
and the sponsor. Observations related to pharmacokinetics,
pharmacodynamics, and any cumulative toxicity observed after
multiple cycles were included in the rationale supporting the
RP2D.
[0226] With regard to safety assessments, AEs were defined as any
untoward medical occurrence in a patient who was administered a
medicinal product that does not necessarily have a causal
relationship with BVD-523, and was coded using the MedDRA coding
dictionary. An SAE was any untoward medical occurrence that
occurred at any dose that resulted in death, was life-threatening,
required inpatient hospitalization or prolongation of existing
hospitalization, or resulted in persistent or significant
disability/incapacity or a congenital anomaly/birth defect. The
severity of AEs were graded according to the National Cancer
Institute Common Terminology Criteria for Adverse Events, Grading
Scale, version 4.
[0227] Safety evaluations were conducted at baseline, on Days 8,
15, 22, 29, 36, and 43, and, in patients who continued treatment,
every 3 weeks or if clinically indicated thereafter. Each
evaluation included a physical examination and clinical laboratory
studies. Electrocardiograms were repeated if clinically significant
and at the discretion of the investigator. The investigators made
judgments regarding whether or not AEs were related to study drug
and followed up until resolution or stabilization, or the AE was
judged to be no longer clinically significant.
[0228] For pharmacokinetic analysis, the pharmacokinetic population
consisted of patients who received at least one dose of BVD-523 and
had evaluable pharmacokinetic data for plasma and/or urine. Blood
samples were collected prior to dosing, and then at 0.5 (.+-.5
min), 1 (.+-.5 min), 2 (.+-.10 min), 4 (.+-.10 min), 6 (.+-.10
min), 8 (.+-.10 min), and 12 (.+-.2 hr) hours on Day 1 (Visit 2;
baseline/initiation of treatment) and Day 15 (Visit 4; at steady
state) after the morning dose. On Day 22, prior to dose
administration, a final blood sample was collected for
pharmacokinetic analyses. Urine samples were collected predose and
at the 1- to 6-hour and 6- to 12-.+-.2-hour intervals postdose on
Days 1 and 15. Plasma and urine samples were analyzed for BVD-523
and metabolites using validated LC/MS/MS methods. Standard
pharmacokinetic parameters were obtained using Phoenix WinNonlin
(Pharsight) with a noncompartmental method. Relationship between
dose and exposure was calculated using standard least-squares
regression analysis.
[0229] For pharmacodynamic confirmation of target inhibition by
BVD-523, targeted ERK inhibition by BVD-523 was determined by
examining pRSK as a target biomarker in human whole blood samples
obtained from patients with advanced solid tumors (N=27) who had
received different doses of BVD-523 (10-900 mg BID) during the
phase I study. The activity of BVD-523 from 4 timepoints (baseline
predose, baseline 4 hours postdose, Day 15 predose, and Day 15 4
hours postdose) was expressed as a percent activity (pRSK) of
PMA-stimulated blood incubated with BVD-523.
[0230] For measurement of antitumor response, tumor measurements
based on physical examination occurred at baseline and on the first
day of each treatment cycle. CT and other assessments were made
every 2 to 3 cycles. Findings were assessed in accordance with
RECIST v1.1: CR was defined as disappearance of all target lesions;
PR was defined as a .gtoreq.30% decrease in the sum of the longest
diameters of target lesions, taking baseline measurements as
reference; stable disease was defined as being of neither
sufficient shrinkage to qualify for PR nor sufficient increase to
qualify for progressive disease, taking as reference the baseline
measurement. Metabolic response was assessed by visualizing tumor
uptake of .sup.18F-glucose via .sup.18F-FDG-PET scanning prior to
receiving the first dose of BVD-523 and at Day 15 (Visit 4).
Example 2
Dose Escalation and Proliferation Assays
Month 1
Dose Escalation Progress--Month 1
[0231] A375 cells were dose escalated using BVD-523, dabrafenib,
and trametinib either as single agents or in combination. Doses
were increased in small increments during the first month. Other
than a marked reduction in growth rate, cells generally tolerated
the escalations well and the doses were planned to be more
aggressively escalated using larger increments in month 2. FIG.
1A-FIG. 1C show month 1 progress for the dose escalation
studies.
Proliferation Assay Results--Month 1
[0232] Proliferation assays were performed to assess the response
of the escalated cells lines vs. parental cell line, to BVD-523,
dabrafenib, and trametinib treatments.
[0233] FIG. 2A-FIG. 2H show normalized and raw proliferation assay
results from month 1 of the studies. Note that differences in max
signals in DMSO controls between different treatments (FIG. 2D,
FIG. 2F, and FIG. 2H) suggest differential growth rates between
treatments. These differences may influence the responses of lines
to inhibitors in the proliferation assays.
[0234] Table 10 shows IC.sub.50 data for month 1 of the
studies.
TABLE-US-00010 TABLE 10 IC.sub.50 Data--Month 1 Cell Line, Relative
IC.sub.50 (nM) BVD- Compound Par* Tram Dab 523 Dab/Tram Dab/523
Tram/523 Dabrafenib 6 29 about 8 58 68 11 161 Trametinib 0.5 2.2
2.5 0.7 3.9 3.1 2.5 BVD-523 189 335 350 268 300 412 263 Paclitaxel
2.2 3.0 3.3 3.4 3.5 3.4 3.4 *Par = Parental cell line
[0235] There were early hints that cells grown in the presence of
escalating doses of dabrafenib or trametinib, either as single
agents or in combinations, were exhibiting decreased responses to
these two agents in proliferation assays.
[0236] In the early stages of month 2, the growth rate of cells in
the dabrafenib only treatment notably increased relative to the
early stages of month 1. This enabled an increased rate of
progression and suggested that resistance was becoming
apparent.
Example 3
Dose Escalation and Proliferation Assays
Month 2
Dose Escalation Progress--Month 2
[0237] The second month of studies saw most treatments move into a
phase where doses were increased in greater increments (1.5-fold)
compared to the initial gentle escalation phase. The single agent
escalation of dabrafenib and trametinib was quickest, with cells
growing in concentrations equivalent to 100.times. parental cell
IC.sub.50 (FIG. 3A and FIG. 3B). The single agent escalation of
BVD-523 progressed more slowly compared to dabrafenib and
trametinib (FIG. 3C). See FIG. 3D for a comparison of the single
agent escalations. BVD-523 escalated cells had a more "fragile"
appearance and there was a greater number of floating cells
compared to the dabrafenib and trametinib escalated
populations.
[0238] The combined agent escalations progressed more slowly than
the single agent treatments. The BVD-523/trametinib combination was
particularly effective in preventing cells from progressing.
Proliferation Assay Results--Month 2
[0239] Proliferation assays on single agent escalated dabrafenib
and trametinib cell populations revealed modest shifts in the dose
response curves, suggesting that an additional period of escalation
would be beneficial to further enrich for resistant cells.
Interestingly, in the proliferations assay, there was evidence to
suggest that cells exposed to BVD-523 grew less well upon inhibitor
withdrawal, perhaps indicating a level of addiction.
[0240] FIG. 4A-FIG. 4H show normalized and raw proliferation assay
results from month 2 of the studies. Note that differences in max
signals in DMSO controls between different treatments (FIG. 4D,
FIG. 4F, and FIG. 4H) suggest differential growth rates between
treatments. These differences may influence the responses of lines
to inhibitors in the proliferation assays.
[0241] FIG. 5A-FIG. 5H show normalized and raw proliferation assay
results from month 2 of the studies with a focus on parental and
BVD-523 line data only.
TABLE-US-00011 TABLE 11 IC.sub.50 Data--Month 2 Cell Line, Relative
IC.sub.50 (nM) BVD- Compound Par* Tra Dab 523 Dab/Tram Dab/523
Tram/523 Dabrafenib 4.1 6.2 11.5 697 256 218 68 Trametinib 0.4 0.7
1.1 24.3 12.6 6.2 4.6 BVD-523 187 252 284 1706 561 678 435
Paclitaxel 3.7 8.9 1.9 6.5 4.7 4.2 8.9 *Par = Parental cell
line
Example 4
Dose Escalation and Proliferation Assays
Month 3
Dose Escalation Progress--Month 3
[0242] FIG. 6A-FIG. 6C show single and combination agent escalation
for month 3 of the studies. FIG. 6D shows a comparison of single
agent escalations.
Proliferation Assay Results--Month 3
[0243] FIG. 7 shows an assessment of growth during the
proliferation assay in DMSO control wells. FIG. 8A-FIG. 8D show
results from month 3 of the studies. FIG. 9A-FIG. 9D show results
from month 3 of the studies with a focus on single treatment cell
lines.
[0244] Table 12 shows IC.sub.50 data for month 3 of the studies.
Relative IC.sub.50s were determined from 4-parameter curve fits in
Prism. IC.sub.50 values were not determined for the cell line
escalated with trametinib due to a lack of growth during the assay
(ND: not done).
TABLE-US-00012 TABLE 12 IC.sub.50 Data--Month 3 Cell Line, Relative
IC.sub.50 (nM) BVD- Compound Par* Tram Dab 523 Dab/Tram Dab/523
Tram/523 Dabrafenib 2.1 ND 2.5 18.4 17.9 337 73 Trametinib 0.2 ND
0.4 1.7 2.7 90 11.2 BVD-523 129 ND 198 433 323 1151 296 Paclitaxel
1.9 ND 1.9 6.5 4.7 4.2 8.9 *Par = Parental cell line
[0245] FIG. 19 shows single and combination agent escalation for
month 3 of the studies. Cell line variants were obtained that could
grow in the presence of dabrafenib or trametinib at concentrations
greater than 100 times the IC.sub.50 of these agents in parental
A375 cell. In comparison, cell lines resistant to BVD-523 could
only be maintained in less than 10.times. of parental IC.sub.50
concentration. Sensitivity testing suggested dabrafenib and
trametinib-resistant cell lines remained relatively sensitive to
BVD-523; the increased IC.sub.50 "shift" for BVD-523 in resistant
cell lines was more modest than those corresponding IC.sub.50
increases following dabrafenib or trametinib treatment. Likewise,
compared to dabrafenib or trametinib treatment, more complete
inhibition of cell growth was observed when resistant cell lines
were treated with BVD-523 at concentrations 10-fold above its
IC.sub.50 in the parental A375 line. In total, patterns of
resistance and cross-sensitivity suggest BVD-523 may remain
effective in settings of acquired resistance.
Example 5
Combination Study Results
[0246] As expected, A375 cells, which carry a BRAF (V600E)
mutation, were sensitive to dabrafenib. Single agent IC.sub.50
values calculated using Alamar Blue (FIG. 10A-FIG. 10E, FIG.
12A-FIG. 12E, and FIG. 14A-FIG. 14E) were generally slightly lower
for Dabrafenib and BVD-523 compared to those derived using
CellTiter-Glo (FIG. 11A-FIG. 11E, FIG. 13A-FIG. 13E, and FIG.
15A-FIG. 15E). Published IC.sub.50 values for Dabrafenib and
Trametinib in a 72 hour CellTiter-Glo assay were 28.+-.16 nM and
5.+-.3 nM respectively (Greger et al., 2012; King et al.,
2013)--the single agent results reported here are consistent with
these values. There was some evidence for a window of synergy in
all treatments. Variation between triplicates was low, however,
there was some evidence of edge effects that likely explains the
apparent enhanced growth observed in some treatments versus the no
drug control (e.g. particularly apparent in the Trametinib/BVD-523
combination). This makes the interpretation of the Bliss analysis
more challenging as in some treatments it may have resulted in the
artefactual enhancement in the level of synergy.
[0247] The combination assays were repeated for A375 cells. Single
agent BVD-523, Trametinib and Dabrafenib potencies were consistent
with those reported in the previous studies disclosed herein.
[0248] In sum, taken together the data show that MEK and BRAF
resistant cells could be overcome by treatment with the ERK
inhibitor, BVD-523.
Example 6
BVD-523 Altered Markers of MAPK Kinase Activity and Effector
Function
[0249] For Western blot studies, HCT116 cells (5.times.10.sup.6)
were seeded into 10 cm dishes in McCoy's 5A plus 10% FBS. A375
cells (2.5.times.10.sup.6) were seeded into 10 cm dishes in DMEM
plus 10% FBS. Cells were allowed to adhere overnight prior to
addition of the indicated amount of test compound (BVD-523) or
vehicle control. Cells were treated for either 4 or 24 hours before
isolation of whole-cell protein lysates, as specified below. Cells
were harvested by trypsinisation, pelleted and snap frozen. Lysates
were prepared with RIPA (Radio-Immunoprecipitation Assay) buffer,
clarified by centrifugation and quantitated by bicinchoninic acid
assay (BCA) assay. 20-50 .mu.g of protein was resolved by SDS-PAGE
electrophoresis, blotted onto PVDF membrane and probed using the
antibodies detailed in Table 13 (for the 4-hour treatment) and
Table 14 (for the 24-hour treatment) below.
TABLE-US-00013 TABLE 13 Antibody Details Incubation/Block Antigen
Size (kDa) Supplier Cat No Dilution Conditions Secondary pRSK1/2 90
Cell 9335 1:1000 o/n 4.degree. C. 5% anti-rabbit pS380 Signaling
BSA pRSK1/2 90 Cell 11989 1:2000 o/n 4.degree. C. 5% anti-rabbit
pS380 Signaling BSA pRSK- 90 Millipore 04-419 1:40000 o/n 4.degree.
C. 5% anti-rabbit T359/5363 BSA Total RSK 90 Cell 9333 1:1000 o/n
4.degree. .C 5% anti-rabbit Signaling BSA pErk 1/2 42/44 Cell 9106S
1:500 o/n 4.degree. C. 5% anti-mouse Signaling milk Total ERK 42/44
Cell 9102 1:2000 o/n 4.degree. C. 5% anti-rabbit Signaling milk
pMEK1/2 45 Cell 9154 1:1000 o/n 4.degree. C. 5% anti-rabbit
Signaling BSA Total MEK 45 Cell 9126 1:1000 o/n 4.degree. C. 5%
anti-rabbit Signaling BSA pS6- 32 Cell 2211S 1:3000 o/n 4.degree.
C. 5% anti-rabbit pS235 Signaling milk Total S6 32 Cell 2217 1:2000
o/n 4.degree. C. 5% anti-rabbit Signaling milk DUSP6 48 Cell 3058S
1:1000 o/n 4.degree. C. 5% anti-rabbit Signaling BSA Total 73 BD
Bio- 610152 1:2000 o/n 4.degree. C. 5% anti-mouse CRAF sciences
milk pCRAF- 73 Cell 9427 1:1000 o/n 4.degree. C. 5% anti-rabbit
Ser338 Signaling BSA pRB 105 Cell 9307 1:2000 o/n 4.degree. C. 5%
anti-rabbit (Ser780) Signaling BSA .beta.-Actin 42 Sigma A5441
1:500,000 o/n 4.degree. C. 5% anti-mouse milk
TABLE-US-00014 TABLE 14 Antibody details Size Incubation/Block
Antigen (kDa) Supplier Cat No Dilution Conditions Secondary pRB 105
Cell 9307 1:2000 o/n 4.degree. C. 5% anti-rabbit (Ser780) Signaling
BSA CCND1 34 Abcam ab6152 1:500 o/n 4.degree. C. 5% anti-mouse milk
Bim-EL 23 Millipore AB17003 1:1000 o/n 4.degree. C. 5% anti-rabbit
BSA Bim-EL 23 Cell 2933 1:1000 o/n 4.degree. C. 5% anti-rabbit
Signaling BSA BCL-xL 30 Cell 2762 1:2000 o/n 4.degree. C. 5%
anti-rabbit Signaling BSA PARP 116/89 Cell 9542 1:1000 o/n
4.degree. C. 5% anti-rabbit Signaling milk Cleaved 17, 19 Cell
9664X 1:1000 o/n 4.degree. C. 5% anti-rabbit Caspase 3 Signaling
milk DUSP6 48 Cell 3058S 1:1000 o/n 4.degree. C. 5% anti-rabbit
Signaling BSA pRSK1/2 90 Cell 9335 1:1000 o/n 4.degree. C. 5%
anti-rabbit pS380 Signaling BSA pRSK1/2 90 Cell 11989 1:2000 o/n
4.degree. C. 5% anti-rabbit pS380 Signaling BSA pRSK- 90 Millipore
04-419 1:40000 o/n 4.degree. C. 5% anti-rabbit T359/5363 BSA Total
RSK 90 Cell 9333 1:1000 o/n 4.degree. C. 5% anti-rabbit Signaling
BSA pErk 1/2 42/44 Cell 9106S 1:500 o/n 4.degree. C. 5% anti-mouse
Signaling milk Total ERK 42/44 Cell 9102 1:2000 o/n 4.degree. C. 5%
anti-rabbit Signaling milk B-Actin 42 Sigma A5441 1:500,000 o/n
4.degree. C. 5% anti-mouse milk
[0250] FIG. 16A-FIG. 16D, FIG. 17A-FIG. 17D, and FIG. 18A-FIG. 18D
show Western blot analyses of cells treated with BVD-523 at various
concentrations for the following: 1) MAPK signaling components in
A375 cells after 4 hours; 2) cell cycle and apoptosis signaling in
A375 24 hours treatment with various amounts of BVD-523; and 3)
MAPK signaling in HCT-116 cells treated for 4 hours. The results
show that acute and prolonged treatment with BVD-523 in RAF and RAS
mutant cancer cells in-vitro affects both substrate phosphorylation
and effector targets of ERK kinases. The concentrations of BVD-523
required to induce these changes is typically in the low micromolar
range.
[0251] Changes in several specific activity markers are noteworthy.
First, the abundance of slowly migrating isoforms of ERK kinase
increase following BVD-523 treatment; modest changes can be
observed acutely, and increase following prolonged treatment. While
this could indicate an increase in enzymatically active,
phosphorylated forms of ERK, it remains noteworthy that multiple
proteins subject to both direct and indirect regulation by ERK
remain "off" following BVD-523 treatment. First, RSK1/2 proteins
exhibit reduced phosphorylation at residues that are strictly
dependent on ERK for protein modification (T359/S363). Second,
BVD-523 treatment induces complex changes in the MAPK feedback
phosphatase, DUSP6: slowly migrating protein isoforms are reduced
following acute treatment, while total protein levels are greatly
reduced following prolonged BVD-523 treatment. Both of these
findings are consistent with reduced activity of ERK kinases, which
control DUSP6 function through both post-translational and
transcriptional mechanisms. Overall, despite increases in cellular
forms of ERK that are typically thought to be active, it appears
likely that cellular ERK enzyme activity is fully inhibited
following either acute or prolonged treatment with BVD-523.
[0252] Consistent with these observations, effector genes that
require MAPK pathway signaling are altered following treatment with
BVD-523. The G1/S cell-cycle apparatus is regulated at both
post-translational and transcriptional levels by MAPK signaling,
and cyclin-D1 protein levels are greatly reduced following
prolonged BVD-523 treatment. Similarly, gene expression and protein
abundance of apoptosis effectors often require intact MAPK
signaling, and total levels of Bim-EL increase following prolonged
BVD-523 treatment. As noted above, however, PARP protein cleavage
and increased apoptosis were not noted in the A375 cell background;
this suggests that additional factors may influence whether changes
in BVD-523/ERK-dependent effector signaling are translated into
definitive events such as cell death and cell cycle arrest.
[0253] Consistent with the cellular activity of BVD-523, marker
analysis suggests that ERK inhibition alters a variety of molecular
signaling events in cancer cells, making them susceptible to both
decreased cell proliferation and survival.
[0254] In sum, FIG. 16A-FIG. 16D, FIG. 17A-FIG. 17D, and FIG.
18A-FIG. 18D show that BVD-523 inhibits the MAPK signaling pathway
and may be more favorable compared to RAF or MEK inhibition in this
setting.
[0255] Finally, properties of BVD-523 may make this a preferred
agent for use as an ERK inhibitor, compared to other agents with a
similar activity. It is known that kinase inhibitor drugs display
unique and specific interactions with their enzyme targets, and
that drug efficacy is strongly influenced by both the mode of
direct inhibition, as well as susceptibility to adaptive changes
that occur following treatment. For example, inhibitors of ABL,
KIT, EGFR and ALK kinases are effective only when their cognate
target is found in active or inactive configurations. Likewise,
certain of these inhibitors are uniquely sensitive to either
secondary genetic mutation, or post-translational adaptive changes,
of the protein target. Finally, RAF inhibitors show differential
potency to RAF kinases present in certain protein complexes and/or
subcellular localizations. In summary, as ERK kinases are similarly
known to exist in diverse, variable, and complex biochemical
states, it appears likely that BVD-523 may interact with and
inhibit these targets in a fashion that is distinct and highly
preferable to other agents.
Example 7
Effects of BVD-523 and Benchmark ERK BRAF and MEK Inhibitors on
Viability and MAPK Signalling Single Agent Proliferation Assay
[0256] Cells were seeded in 96-well plates at the densities
indicated in Table 15 in McCoy's 5A containing 10% FBS and allowed
to adhere overnight prior to addition of compound or vehicle
control. Compounds were prepared from DMSO stocks to give the
desired final concentrations. The final DMSO concentration was
constant at 0.1%. Test compounds were incubated with the cells for
96 h at 37.degree. C., 5% CO.sub.2 in a humidified atmosphere.
CellTiter-Glo.RTM. reagent (Promega, Madison, Wis.) was added
according to manufacturer's instructions and luminescence detected
using the BMG FLUOstar plate reader (BMG Labtech, Ortenberg,
Germany). The average media only background value was deducted and
the data analysed using a 4-parameter logistic equation in GraphPad
Prism (GraphPad Software, La Jolla, Calif.).
Combination Proliferation Assay
[0257] Cells were seeded into triplicate 96-well plates at the
densities indicated in Table 15 in McCoy's 5A containing 10% FBS
and allowed to adhere overnight prior to addition of test compound
or vehicle control. Combinations were tested using a 10.times.8
dose matrix. The final DMSO concentration was constant at 0.2%.
[0258] Test compounds were incubated with the cells for 96 h at
37.degree. C., 5% CO.sub.2 in a humidified atmosphere. Cells were
stained with Hoechst stain and fluorescence detected as described
above. The average media only background value was deducted and the
data analysed.
[0259] Combination interactions across the dose matrix were
determined by the Loewe Additivity and Bliss independence models
using Chalice.TM. Combination Analysis Software (Horizon Discovery
Group, Cambridge, Mass.) as outlined in the user manual (available
at
chalice.horizondiscovery.com/chalice-portal/documentation/analyzer/home.j-
sp). Synergy is determined by comparing the experimentally observed
level of inhibition at each combination point with the value
expected for additivity, which is derived from the single-agent
responses along the edges of the matrix. Potential synergistic
interactions were identified by displaying the calculated excess
inhibition over that predicted as being additive across the dose
matrix as a heat map, and by reporting a quantitative `Synergy
Score` based on the Loewe model. The single agent data derived from
the combination assay plates were presented as dose-response curves
generated in Chalice.TM.
TABLE-US-00015 TABLE 15 Cell Line Seeding Density Seeding density
(cells/well) 96-well 6-Well 10 cm dish Cell Line Proliferation
Western Westerns RKO Parental 1000 .sup. 1 .times. 10.sup.6 2.9
.times. 10.sup.6 RKO MEK1 1250 Not tested Not tested (Q56P/+) Clone
1 RKO MEK1 1000 7.5 .times. 10.sup.5 .sup. 2 .times. 10.sup.6
(Q56P/+) Clone 2
Western Blotting
[0260] Cells were seeded into 6-well plates (Experiment 1) or 10 cm
dishes (Experiment 2) at the densities indicated in Table 15 in
McCoy's 5A containing 10% FBS and allowed to adhere overnight prior
to addition of compound or vehicle control. Test compounds were
added and incubated with the cells for 4 or 24 h at 37.degree. C.,
5% CO.sub.2 in a humidified atmosphere. Cells were harvested by
trypsinisation, pelleted by centrifugation and snap frozen on dry
ice.
[0261] Lysates were prepared using RIPA buffer (50 mM
Tris-hydrochloride, pH 8.0; 150 mM sodium chloride; 1.0% Igepal
CA-630 (NP-40); 0.5% sodium deoxycholate; 0.1% sodium dodecyl
sulphate; 1.times. complete EDTA-free protease inhibitor cocktail
(Roche, Nutley, N.J.; cat 05 892 791 001); 1.times. phosSTOP
phosphatase inhibitor cocktail (Roche Nutley, N.J.; cat. 04 906 837
001)) and clarified by centrifugation at 11,000 rpm for 10 min in a
bench-top centrifuge.
[0262] Total protein in the lysates was quantitated by BCA assay
according to the manufacturer's instructions (Pierce.TM. BCA
Protein Assay Kit; Thermo Scientific, Waltham, Mass.; cat. 23225),
boiled in sample buffer (NuPAGE LDS Sample Buffer; (Invitrogen,
Carlsbad, Calif.; cat. NP0007)) and stored at -80.degree. C.
[0263] Equal amounts of protein (40 .mu.g) were resolved on NuPAGE
4-12% Bis-Tris gels (Invitrogen, Carlsbad, Calif.; cat. WG1402BOX)
and blotted onto PVDF membranes using iBlot gel transfer stacks
(Invitrogen, Carlsbad, Calif.; cat. IB4010-01) on an iBlot gel
transfer device (Invitrogen Carlsbad, Calif.) according to the
manufacturer's instructions.
[0264] Blots were probed using the antibodies and block conditions
detailed in Table 16. Western blots were developed using Pierce.TM.
ECL2 Western blotting substrate (Thermo Scientific, Waltham, Mass.;
cat. 80196) and imaged using a FluorChem M Western blot imager
(ProteinSimple, San Jose, Calif.).
TABLE-US-00016 TABLE 16 Antibodies and Western Blotting Conditions
Size Incubation/block Antigen (kDa) Supplier Cat No Dilution
Conditions Secondary pRSK- 90 Millipore 04-419 1:20000 o/n
4.degree. C. 5% BSA anti-rabbit T359/S353 Total RSK 90 Cell
Signaling 9333 1:1000 o/n: 4.degree. C. 5% BSA anti-rabbit pErk 1/2
42/44 Cell Signaling 9106S 1:500 o/n 4.degree. C. 5% milk
anti-mouse Total ERK 42/44 Cell Signaling 9102 1:2000 o/n 4.degree.
C. 5% milk anti-rabbit pMEK1/2 45 Cell Signaling 9154 1:1000 o/n
4.degree. C. 5% BSA anti-rabbit Total MEK 45 Cell Signaling 9126
1:1000 o/n 4.degree. C. 5% BSA anti-rabbit DUSP6 45 Cell Signaling
3058S 1:1000 o/n 4.degree. C. 5% BSA anti-rabbit pRB (Ser780) 105
Cell Signaling 9307 1:2000 o/n 4.degree. C. 5% BSA anti-rabbit
CCND1 34 Abcam ab6152 1:500 o/n 4.degree. C. 5% milk anti-mouse
B-Actin 42 Sigma A5441 1:100,000 o/n 4.degree. C. 5% milk
anti-mouse Anti-rabbit -- Cell Signaling 7074S 1:2000 1 h room
temp. -- HRP- Block matched to conjugated primary Antibody
secondary Anti-mouse -- Cell Signaling 7076 1:5000 1 h room temp.
-- HRP- Block matched to conjugated primary Antibody secondary
[0265] The MEK1 (Q56P) mutation exemplifies a class of clinically
relevant MEK1/2 activating mutations known to up-regulate the MAPK
pathway and drive acquired resistance to BRAF or MEK
inhibitors.
[0266] This study used a pair of RKO BRAF(V600E) cell lines that
are isogenic for the presence or absence of a MEK1 (Q56P)
activating mutation, to assess the effect that activating MEK
mutations have in response to the novel ERK inhibitor BVD-523
versus other benchmark MAPK inhibitors.
[0267] Effects of on cell viability were assessed by quantitating
cellular ATP levels using CellTiter-Glo.RTM. after 96 h. Single
agent assays demonstrated that the double mutant
BRAF(V600E)::MEK1(Q56P) cells displayed a markedly reduced
sensitivity to inhibition with benchmark clinical BRAF (exemplified
by Dabrafenib) or MEK (exemplified by Trametinib) inhibitors
relative to the parental BRAF(V600E) cells, which demonstrates the
suitability of this isogenic model for recapitulating the acquired
resistance known to be associated with this class of mutation in
the clinic (Table 17).
TABLE-US-00017 TABLE 17 Single Agent IC.sub.50 Values RKO MEK1 RKO
MEK1 Q56P/+ Q56P/+ Compound RKO Parental Cl. 1 Cl. 2 BVD-523 0.20
0.17 0.18 SCH772984 0.04 0.14 0.12 Dabrafenib n.d. n.d. n.d.
Trametinib 0.006 0.093 0.080 Paclitaxel 0.002 0.002 0.002 n.d.--not
determined, only a partial dose response achieved
[0268] In contrast, response to BVD-523 was identical in both the
parental and double mutant cells, indicating that BVD-523 is not
susceptible to this mechanism of acquired resistance.
[0269] These results were identical in two independently derived
double mutant BRAF(V600E)::MEK1(Q56P) cell line clones confirming
that these differences in response versus the parental cells were
specifically related to the presence of the MEK1 mutation rather
than an unrelated clonal artifact (FIG. 22A-FIG. 22E). Similar
results were also observed with a second mechanistically distinct
benchmark ERK inhibitor (SCH772984), which supports the notion that
these observations are specifically related to inhibition of ERK
and not due to an off-target effect.
[0270] The effect of combining BVD-523 with a BRAF inhibitor
(exemplified by Dabrafenib) was also assessed in these cell lines
across a matrix of concentrations using the Loewe Addivity or Bliss
Independence models with Horizon's Chalice.TM. combination analysis
software (FIG. 23-FIG. 23O and FIG. 24A-FIG. 24O). The presence of
potentially synergistic interactions was then assessed by
displaying the calculated excess inhibition over that predicted as
being additive across the dose matrix as a heat map, and by
calculating a `Volume Score` that shows whether the overall
response to a combination is synergistic (positive values),
antagonistic (negative values) or additive (.about.0).
[0271] The results suggest that the BVD-523::Dabrafenib combination
was mainly additive in the parental and mutant cell line. In
contrast, the combination of a MEK inhibitor (trametinib) plus
Dabrafenib, while being mostly additive in the parental cell line,
showed strong synergy in the double mutant BRAF(V600E)::MEK1(Q56P)
cell line (FIG. 25A-FIG. 25O). Loewe Volumes, Bliss Volumes and
Synergy scores for the combinations tested are shown in Tables
18-20, respectively and are shown graphed in FIG. 26A-FIG. 26C.
TABLE-US-00018 TABLE 18 Loewe Volumes RKO MEK1 RKO MEK1 (Q56P) -
(Q56P) - RKO Parental Clone 1 Clone 2 BVD-523 .times. Dabrafenib
3.54 2.88 2.35 Dabrafenib .times. SCH772984 5.2 6.79 6.14
Dabrafenib .times. Trametinib 5.68 12.6 11.6
TABLE-US-00019 TABLE 19 Bliss Volumes RKO MEK1 RKO MEK1 (Q56P) -
(Q56P) - RKO Parental Clone 1 Clone 2 BVB-523 .times. Dabrafenib
-0.894 0.527 1.42 Dabrafenib .times. SCH772984 0.209 4.3 5.07
Dabrafenib .times. Trametinib 0.353 10.8 9.87
TABLE-US-00020 TABLE 20 Synergy Scores RKO MEK1 RKO MEK1 (Q56P) -
(Q56P) - RKO Parental Clone 1 Clone 2 BVD-523 .times. Dabrafenib
3.18 2.31 1.77 Dabrafenib .times. SCH772984 4.56 5.57 4.36
Dabrafenib .times. Trametinib 5.58 11 9.83
[0272] Effects on MAPK pathway signally was assessed by Western
blotting. The levels of basal ERK phosphorylation (DMSO samples)
was markedly up-regulated in the MEK1(Q56P)-expressing line
relative to parental further confirming that this isogenic model
faithfully recapitulates the expected phenotype for the expression
of MEK activating acquired resistance mutations.
[0273] In the parental BRAF(V600E) RKO cells, a reduced level of
RSK1/2 phosphorylation is observed following acute treatment with
RAF, MEK and ERK kinase inhibitors at pharmacologically active
concentrations. In contrast, isogenic, double mutant
BRAFV600E::MEK1Q56P cells do not exhibit reduced RSK
phosphorylation following BRAF or MEK inhibitor treatment, while
BVD-523 remains effective at similar concentrations (FIG. 27A-FIG.
27I). The dotted lines indicate that the trametinib-treated samples
(plus matched DMSO control) and blots are derived from a separate
experiment to the BRAFi and BVD-523 treated samples.
[0274] Changes in effector gene signaling consistent with cell
growth inhibition patterns are observed following prolonged
inhibitor treatment. In parental RKO lines, a reduced level of
phosphorylated pRB is observed following prolonged MEK and ERK
inhibitor treatment. At the level of pRB modulation, MEK1 mutant
lines appear insensitive to low concentration MEK inhibitor
treatment, while higher concentrations remain effective.
Critically, BVD-523 potency against pRB activity does not appear to
be strongly affected by MEK mutation. Surprisingly, RAF inhibitor
treatment does not affect pRB status, despite potent inhibition of
upstream signaling, in both parental and MEK mutant
backgrounds.
[0275] In summary, these results show that BVD-523 is not
susceptible to acquired resistance driven by MEK activating
mutations such as MEK1 (Q56P). In addition they suggest that in
combination the interactions between BVD-523 and BRAFi (exemplified
by Dabrafenib) are additive irrespective of the presence of a MEK
activating mutation.
Example 8
Combination Interactions Between ERK Inhibitors
[0276] RAF mutant melanoma cell line A375 cells were cultured in
DMEM with 10% FBS and seeded into triplicate 96-well plates at an
initial density of 2000 cells per well. Combination interactions
between ERK inhibitors BVD-523 and SCH772984 were analized after 72
hours as described above in Example 4. Viability was determined
using CellTiter-Glo.RTM. reagent (Promega, Madison, Wis.) according
to manufacturer's instructions and luminescence was detected using
the BMG FLUOstar plate reader (BMG Labtech, Ortenberg,
Germany).
[0277] Visualization of the Loewe and Bliss `excess inhibition`
heat maps suggested that the combination of BVD-523 and SCH772984
was mainly additive with windows of potential synergy in mid-range
doses (FIG. 28A-FIG. 28E).
[0278] In summary, these results suggest that interactions between
BVD-523 and SCH772984 are at least additive, and in some cases
synergistic.
Example 9
Targeting the MAPK Signaling Pathway in Cancer
Promising Activity with the Novel Selective ERK1/2 Inhibitor
BVD-523 (Ulixertinib)
[0279] Treatment strategies for cancer have evolved from classic
cytotoxic-based approaches to agents that counteract the effects of
genetic lesions that drive aberrant signaling essential to tumor
proliferation and survival. For example, the ERK module of the
mitogen-activated protein kinase (MAPK) signaling cascade
(RAS-RAF-MEK-ERK) (Cargnello and Rouxx 2011) can be engaged by
several receptor tyrosine kinases (e.g., EGFR and ErbB-2) in
addition to constitutively activated mutations of pathway
components such as RAS and BRAF (Gollob et al. 2006). Through
aberrant activation of ERK signaling, genetic alterations in RAS or
BRAF result in rapid tumor growth, increased cell survival, and
resistance to apoptosis (Poulikakos et al. 2011, Corcoran et al.
2010, Nazarian et al. 2010, Shi et al. 2014, Wagle et al. 2011).
Activating mutations of RAS family members KRAS and NRAS are found
in .about.30% of all human cancers, with particularly high
incidence in pancreatic (Kanda et al. 2012) and colorectal cancer
(Arrington et al. 2014). Constitutively activating mutations in the
BRAF gene that normally encodes for valine at amino acid 600 have
been observed in melanoma, thyroid carcinoma, colorectal cancer,
and non-small cell lung cancer (Hall et al. 2014). Cancers bearing
genetic mutations that result in changes of the downstream
components ERK and MEK have also been reported (Ojesina et al.
2014, Arcila et al. 2015). Alterations that activate the MAPK
pathway are also common in the setting of resistance to targeted
therapies (Groenendijk et al. 2014). Thus, targeting the MAPK
pathway terminal master kinases (ERK1/2) is a promising strategy
for tumors harboring such pathway activating alterations (e.g.,
BRAF, NRAS, and KRAS).
[0280] Three MAPK pathway-targeting drugs have been approved by the
US Food and Drug Administration (FDA) for single-agent treatment of
nonresectable or metastatic cutaneous melanoma with BRAF.sup.V600
mutations: the BRAF inhibitors vemurafenib and dabrafenib and the
MEK inhibitor trametinib. Furthermore, the combination of
dabrafenib and trametinib is also approved in this indication
(Queirolo et al. 2015 and Massey et al. 2015). An additional MEK
inhibitor, cobimetinib, is approved in this indication as part of a
combination regimen with BRAF inhibitors. Clinical experience with
these drugs validates the MAPK pathway as a therapeutic target. In
phase III trials of patients with BRAF.sup.V600-mutant melanoma,
the single agents vemurafenib and dabrafenib demonstrated superior
response rates (approximately 50% vs. 5-19%) and median
progression-free survival (PFS, 5.1-5.3 months vs. 1.6-2.7 months)
over cytotoxic chemotherapy (dacarbazine) (Chapman et al. 2011 and
Hauschild et al. 2012). Furthermore, clinical use of concomitant
BRAF-plus MEK-targeted therapies has demonstrated that simultaneous
targeting of different nodes in the MAPK pathway can enhance the
magnitude and duration of response. First-line use of BRAF plus
MEK-targeted agents (dabrafenib/trametinib or
cobimetinib/vemurafenib) further improved median overall survival
compared with single-agent BRAF inhibition (Robert et al. 2015,
Long et al. 2015, Larkin et al. 2014). Thus, combined
BRAF-/MEK-targeted therapy is a valuable treatment option for
patients with metastatic melanoma with BRAF.sup.V600 mutations.
[0281] Despite improvements in clinical outcomes seen with
BRAF-/MEK-inhibitor combination therapies, durable benefit is
limited by the eventual development of acquired resistance and
subsequent disease progression, with median PFS ranging from
approximately 9 to 11 months. (Robert et al. 2015, Long et al.
2015, Larkin et al. 2014, and Flaherty et al. 2012). Genetic
mechanisms of acquired resistance to single-agent BRAF inhibition
have been intensely studied, and identification of resistance
mechanisms include splice variants of BRAF (Poulikakos et al.
2011), BRAF.sup.V600E amplification (Corcoran et al. 2010), MEK
mutations (Wagle et al. 2014), NRAS mutations, and RTK activation
(Nazarian et al. 2010 and Shi et al. 2014). Resistance mechanisms
in the setting of BRAF-/MEK-inhibitor combination therapy are
beginning to emerge and mirror that of BRAF single-agent resistance
(Wagle et al. 2014 and Long et al. 2014). These genetic events all
share in common the ability to reactivate ERK signaling. Indeed,
reactivated MAPK pathway signaling as measured by ERK
transcriptional targets is common in tumor biopsies from BRAF
inhibitor-resistant patients (Rizos et al. 2014). Furthermore,
ERK1/2 reactivation has been observed in the absence of a genetic
mechanism of resistance (Carlino et al. 2015). Therefore, the quest
to achieve durable clinical benefit has led researchers to focus on
evaluating additional agents that target the downstream MAPK
components ERK1/2. Inhibiting ERK may provide important clinical
benefit to patients with acquired resistance to BRAF/MEK
inhibition. ERK family kinases have shown promise as therapeutic
targets in preclinical cancer models, including those cancers
resistant to BRAF or MEK inhibitors (Morris et al. 2013 and
Hatzivassiliou et al. 2012). However, the potential use of such
ERK1/2 inhibitors expands beyond acquired-resistance in
melanoma.
[0282] Targeting ERK1/2 is a rational strategy in any tumor type
harboring known drivers of MAPK, not only BRAF/MEK therapy-relapsed
patients. As ERK1 and ERK2 reside downstream in the pathway, they
represent a particularly attractive treatment strategy within the
MAPK cascade that may avoid upstream resistance mechanisms. Here,
preclinical characterization of BVD-523 (ulixertinib) in models of
MAPK pathway-dependent cancers is reported, including drug-naive
and BRAF/MEK therapy acquired-resistant models. Results of a phase
I dose-finding study of BVD-523 are included as a companion
publication in this journal. See, Examples 17-24.
[0283] In the present invention, BVD-523 was shown to be a potent,
highly selective, reversible, small molecule ATP-competitive
inhibitor of ERK1/2 with in vitro and in vivo anticancer
activity.
[0284] BVD-523 (ulixertinib) was identified and characterized as a
novel, reversible, ATP-competitive ERK1/2 inhibitor with high
potency and ERK1/2 selectivity. BVD-523 caused reduced
proliferation and enhanced caspase activity, most notably in cells
harboring MAPK (RAS-RAF-MEK) pathway mutations. In in vivo
BRAF.sup.V600E xenograft studies, BVD-523 showed dose-dependent
growth inhibition and tumor regressions. Interestingly, BVD-523
inhibited phosphorylation of target substrates despite increased
phosphorylation of ERK1/2. BVD-523 also demonstrated antitumor
activity in models of acquired resistance to single-agent and
combination BRAF/MEK targeted therapy. Synergistic
antiproliferative effects in a BRAF.sup.V600E-mutant melanoma cell
line xenograph model were also demonstrated when BVD-523 was used
in combination with BRAF inhibition. These studies suggest that
BVD-523 holds promise as a treatment for ERK-dependent cancers,
including those whose tumors have acquired resistance to other
treatments targeting upstream nodes of the MAPK pathway.
Example 10
Discovery and Initial Characterization of a Novel ERK1/2 Inhibitor,
BVD-523 (Ulixertinib)
[0285] Following extensive optimization of leads originally
identified using a high-throughput, small-molecule screen (Aronov
et al. 2009), a novel adenosine triphosphate (ATP)-competitive
ERK1/2 inhibitor, BVD-523 (ulixertinib) was identified (FIG. 29 A).
BVD-523 is a potent ERK inhibitor with a K.sub.i of 0.04.+-.0.02 nM
against ERK2. It was shown to be a reversible, competitive
inhibitor of ATP, as the IC.sub.50 values for ERK2 inhibition
increased linearly with increasing ATP concentration (FIG. 29B and
FIG. 29C). The IC.sub.50 remained nearly constant for incubation
times 210 minutes, suggesting rapid equilibrium and binding of
BVD-523 with ERK2 (FIG. 29D). BVD-523 is also a tight-binding
inhibitor of recombinant ERK1 (Rudolph et al. 2015), exhibiting a
K.sub.i of <0.3 nM.
[0286] Binding of BVD-523 to ERK2 was demonstrated using
calorimetric studies and compared to data generated using the ERK
inhibitors SCH772984 and pyrazolylpyrrole (Arovov et al. 2007). All
compounds bound and stabilized inactive ERK2 with increasing
concentration, as indicated by positive .DELTA.Tm values (FIG.
29E). The 10- to 15-degree change in .DELTA.Tm observed with
BVD-523 and SCH-772984 is consistent with compounds that have
low-nanomolar binding affinities (Fedorov et al. 2012). BVD-523
demonstrated a strong binding affinity to both phosphorylated
active ERK2 (pERK2) and inactive ERK2 (FIG. 29F). A stronger
affinity to pERK2 compared with inactive ERK2 was observed. BVD-523
did not interact with the negative control protein p38a MAP kinase
(FIG. 29F).
[0287] BVD-523 demonstrated excellent ERK1/2 kinase selectivity
based on biochemical counter-screens against 75 kinases in addition
to ERK1 and ERK2. The ATP concentrations were approximately equal
to the K.sub.m in all assays. Kinases inhibited to greater than 50%
by 2 .mu.M BVD-523 were retested to generate K.sub.i values (or
apparent Ki; Table 21). Twelve of the 14 kinases had a K.sub.i of
<1 .mu.M. The selectivity of BVD-523 for ERK2 was >7000-fold
for all kinases tested except ERK1, which was inhibited with a Ki
of <0.3 nM (10-fold). Therefore, BVD-523 is a highly potent and
selective inhibitor of ERK1/2.
TABLE-US-00021 TABLE 21 BVD-523 displays selectivity for ERK1 and
ERK2 kinases. Kinase Ki (.mu.M) CDK1/cyclinB 0.07.sup.a
CDK2/cyclinA 0.36 CDK5/p35 0.09.sup.a CDK6/cycinD3 0.09.sup.a ERK1
0.0003 ERK2 0.00004 GSK3b 0.32 JNK2.alpha. 0.65.sup.a JNK3 1.3
P38.gamma. 0.45.sup.a P38.delta. 0.24.sup.a ROCKI 11.1 ROCKII
0.27.sup.a RSK3 0.45 .sup.aApparent. <50% inhibition at 2 .mu.M:
ABL, AKT3, AMPK, AUR1, AUR2, AXL, BLK, CAMKII, CAMKIV, CHK1, CHK2,
CK1, CK2, CSK, EGFR, EPHB4, FES, FGFR3, FLT3, FYN, IGF1R,
IKK.alpha., IKK.beta., IKKi, IRAK4, IRTK, ITK, JAK3, JNK1.alpha. 1,
KDR, LCK, LYN, cMET, MKK4, MKK6, MKK7.beta., MLK2, MSK1, MST2, NAK,
NEK2, p38.alpha., p38.beta., p70S6K, PAK2, PDGFR.alpha., PDK1, PKA,
PKC.alpha., PKC.beta. II, PKC.gamma., PKCi, PKC.theta., PRAK, PRK2,
cRAF, SGK, SRC, SYK, TAK1, TIE2, ZAP70
Example 11
BVD-523 Preferentially Inhibits Cellular Proliferation and Enhances
Caspase-3/7 Activity In Vitro in Cancer Cell Lines with MAPK
Pathway-Activating Mutations
[0288] BVD-523 cellular activity was assessed in a panel of
approximately 1,000 cancer cell lines of various lineages and
genetic backgrounds (FIG. 30A and Table 22). Cell lines were
classified as MAPK wild type (wt) or mutant depending on the
absence or presence of mutations in RAS family members and BRAF.
Although some MAPK-wt cell lines were sensitive to BVD-523,
generally BVD-523 inhibited proliferation preferentially in cells
with MAPK pathway alterations.
[0289] Next, the growth and survival impact of BVD-523 treatment on
sensitive cells was characterized. Fluorescence activated cell
sorting (FACS) analysis was performed on BRAF.sup.V600E-mutant
melanoma cell line UACC-62 following treatment with BVD-523 at 500
nM or 2000 nM for 24 hours. Treated cells were arrested in the G1
phase of the cell cycle in a concentration-dependent manner (FIG.
30B).
[0290] In addition, caspase-3/7 activity was analyzed as a measure
of apoptosis in multiple human cancer cell lines. A concentration-
and cell-line-dependent increase in caspase 3/7 was observed
following treatment with BVD-523 for 72 hours (FIG. 30C). BVD-523
treatment resulted in pronounced caspase-3/7 induction in a subset
of MAPK-activated cell lines harboring a BRAF.sup.V600 mutation
(A375, WM266, and LS411N). This is consistent with earlier
observations for preferential inhibition of proliferation by
BVD-523 in MAPK pathway-mutant cancer cell lines (FIG. 30A).
[0291] To further characterize the mechanism of action and effects
on signaling elicited by BVD-523, the levels of various effector
and MAPK-related proteins were assessed in BVD-523-treated
BRAF.sup.V600E-mutant A375 melanoma cells (FIG. 30D).
Phospho-ERK1/2 levels increased in a concentration-dependent manner
after 4 and 24 hours of BVD-523 treatment. Despite prominent
concentration-dependent increases in pERK1/2 observed with 2 .mu.M
BVD-523 treatment, phosphorylation of the ERK1/2 target RSK1/2 was
reduced at both 4 and 24 hours, which is consistent with sustained
inhibition. Total protein levels of DUSP6, a distal marker of
ERK1/2 activity, were also attenuated at 4 and 24 hours. Following
24 hours of treatment with BVD-523, the apoptotic marker BIM-EL
increased in a dose-dependent manner, while cyclin D-1 and pRB was
attenuated at 2 .mu.M. All effects are consistent with on-target
ERK1/2 inhibition.
TABLE-US-00022 TABLE 22 Cell Compound Fitted Viability ratio
Barcode Organ ID Cell Line No MGH_IC50 20 uM 10 uM 5 uM 2.5 uM 1.25
uM 0.625 uM 0.3125 uM 0.15625 uM 0.078125 uM 026_8049_00277140
Biliary Tract 8049 ETK-1 456 3.525905 0.587 0.805 0.874 0.9304
0.8796 0.954 1.0285 1.094 0.9918 026_664_00277150 Biliary Tract 664
HuCCT1 456 3.600435 0.663 0.734 0.847 0.9661 0.9652 1.029 0.9656
1.0172 0.9981 026_653_00278500 Biliary Tract 653 EGI-1 456 4.229085
0.693 0.686 0.74 0.7843 0.8546 0.889 0.967 0.9286 0.9525
026_8204_00278540 Biliary Tract 8204 TGBC24TKB 456 5.609877 0.768
0.875 0.826 0.8122 0.8629 0.862 0.8909 0.9353 0.9353
026_8188_00293390 Biliary Tract 8188 TGBC1TKB 456 6.179372 0.915
0.929 0.913 0.9808 0.9201 1.144 1.0128 0.9048 0.9496
026_330_00278580 Bone 330 H-EMC-SS 456 0.038629 0.36 0.392 0.383
0.4615 0.437 0.566 0.7772 0.9442 0.9662 026_8047_00283120 Bone 8047
ES7 456 1.846677 0.515 0.521 0.539 0.5362 0.7569 0.769 0.8353
0.9371 0.9503 026_8053_00287650 Bone 8053 EW-13 456 2.197657 0.333
0.545 0.665 0.8165 0.9099 0.962 0.9571 0.9945 1.0406
026_8227_00288230 Bone 8227 CADO-ES1 456 2.29467 0.359 0.554 0.585
1.0389 0.9034 0.952 1.1264 1.2671 1.0023 026_8050_00279380 Bone
8050 EW-1 456 2.409222 0.487 0.555 0.593 0.6879 0.6908 0.788 0.8057
0.892 0.9331 026_306_00278530 Bone 306 SK-ES-1 456 2.4607 0.527
0.556 0.577 0.682 0.673 0.855 0.8706 0.8641 0.8867 026_305_00277180
Bone 305 U-2 OS 456 2.847932 0.145 0.599 0.657 0.7441 0.8231 0.793
0.8449 0.9969 0.8925 026_337_00283440 Bone 337 HuO9 456 2.916396
0.448 0.696 0.939 0.8026 0.8478 0.903 0.9796 0.8682 1.0954
26_8227_00304340 Bone 8227 CADO-ES1 456 2.975471 0.542 0.666 0.884
0.8971 0.9918 0.974 1.0247 1.0031 1.0728 026_8043_00283110 Bone
8043 ES1 456 2.981717 0.543 0.663 0.705 0.7865 0.811 0.835 0.7995
0.9278 0.8292 026_8142_00282550 Bone 8142 NOS-1 456 3.203574 0.547
0.775 0.825 0.892 0.7556 0.894 0.8477 1.1038 0.9881
026_8055_00290580 Bone 8055 EW-18 456 3.311765 0.638 0.688 0.718
1.1012 0.9293 0.993 1.0646 1.1078 1.0895 026_8058_00293350 Bone
8058 EW-3 456 3.365484 0.715 0.595 0.72 0.7195 0.8774 0.8 0.8628
0.9101 1.2148 026_339_00277160 Bone 339 NY 456 3.400937 0.59 0.789
0.875 0.9222 0.9594 0.975 0.933 1.1433 0.9637 026_8165_00287690
Bone 8165 SK-PN-DW 456 3.476926 0.621 0.812 0.933 0.9706 0.9188
0.991 1.0026 1.0012 1.0432 026_326_00282540 Bone 326 MHH-ES-1 456
3.524605 0.618 0.719 0.824 0.8235 0.8745 0.966 1.1187 1.0416 0.913
026_8048_00279370 Bone 8048 ES8 456 3.530755 0.572 0.698 0.765
0.8053 0.8255 0.912 0.931 0.9069 0.9095 026_331_00278590 Bone 331
HOS 456 3.602447 0.53 0.798 0.749 0.7609 0.8727 0.839 0.8786 0.8236
0.972 026_8045_00282660 Bone 8045 ES5 456 3.665641 0.704 0.653
0.874 0.8827 0.7407 0.913 0.8294 1.0585 1.162 026_8059_00283090
Bone 8059 EW-7 456 3.827217 0.662 0.72 0.76 0.831 0.8857 0.95
0.9309 0.9266 0.9963 026_8201_00282520 Bone 8201 ES3 456 3.982944
0.707 0.721 0.778 0.8414 0.8385 0.989 0.9588 1.0248 1.0261
026_8056_00314310 Bone 8056 EW-22 456 4.196343 0.701 0.827 0.819
0.8824 0.9413 0.894 0.9848 1.0221 1.1139 026_329_00282700 Bone 329
G-292 Clone A141B1 456 4.264519 0.705 0.918 0.893 0.949 1.0435
1.016 0.9814 0.9215 1.1209 026_324_00278550 Bone 324 CAL-72 456
4.286956 0.731 0.942 0.942 0.9262 0.9863 0.966 0.9797 0.9547 1.0113
026_304_00283460 Bone 304 Saos-2 456 4.597501 0.742 0.784 0.9
0.8765 0.9306 0.926 0.9516 0.95 1.0352 026_325_00283060 Bone 325
CAL-78 456 4.650689 0.727 0.913 0.918 0.9149 0.8999 0.897 0.9065
1.0242 1.0181 026_1138_00278560 Bone 1138 CS1 456 4.765346 0.744
0.848 0.864 0.8601 0.8818 1.016 1.0005 0.9749 0.977
026_8162_00282560 Bone 8162 SJSA-1 456 4.867747 0.802 0.803 0.805
0.7765 0.762 0.974 1.0651 1.0412 1.0623 026_336_00283430 Bone 336
HuO-3N1 456 4.900372 0.724 0.86 0.806 0.8427 0.8308 0.87 0.8972
0.9772 1.0073 026_328_00278600 Bone 328 TC-71 456 5.117725 0.844
0.939 0.97 1.0221 0.9857 1.072 1.023 1.0842 1.0771
026_8054_00282530 Bone 8054 EW-16 456 5.180986 0.835 0.775 0.925
0.8021 0.8155 0.934 0.9339 1.058 1.1414 026_335_00308220 Bone 335
MG-63 456 5.257203 0.884 0.841 0.852 0.8745 1.0185 1.081 1.1007
1.0325 1.0709 026_1241_00283070 Bone 1241 CHSA8926 456 5.394244
0.883 0.851 0.947 0.9788 0.9327 1.019 1.0879 1.0294 1.119
026_8044_00279340 Bone 8044 ES4 456 6.158016 0.822 0.876 0.92
0.8612 0.8592 0.873 0.8969 0.9914 0.9632 026_8057_00283080 Bone
8057 EW-24 456 6.273162 0.927 0.899 0.97 0.9817 0.9704 1.012 0.9822
0.9596 0.9793 026_8051_00285230 Bone 8051 EW-11 456 6.340509 0.924
1.008 0.846 0.9309 1.047 0.868 0.9762 1.0834 1.048
026_8046_00279351 Bone 8046 ES6 456 6.745328 0.882 1.065 1.046
1.0203 1.0617 0.85 0.8881 1.0547 0.9578 026_8146_00285140 Brain
8146 ONS-76 456 1.017095 0.362 0.369 0.387 0.52 0.6146 0.733 0.8773
0.9176 1.1016 026_8009_00285111 Brain 8009 AM-38 456 2.859683 0.545
0.617 0.664 0.5881 0.5592 0.692 0.826 0.9522 1.078
026_8091_00285281 Brain 8091 KS-1 456 2.979142 0.617 0.59 0.62
0.7057 0.7844 0.875 0.9025 1.0584 1.1343 026_388_00285240 Brain 388
MOG-G-CCM 456 3.029922 0.505 0.754 0.852 0.977 0.9597 0.989 1.1211
1.0361 1.0747 026_352_00283150 Brain 352 LN-229 456 3.053877 0.552
0.647 0.662 0.7206 0.8119 0.932 0.9285 1.0381 1.0748
026_8214_00290680 Brain 8214 YH-13 456 3.068585 0.558 0.628 0.773
1.0443 0.9614 1.113 1.1139 1.093 1.2619 026_8214_00288290 Brain
8214 YH-13 456 3.531592 0.627 0.651 0.866 0.8387 0.8073 0.922 1.077
1.0619 1.1182 026_358_00293700 Brain 358 D283 Med 456 3.551231 0.64
0.743 0.916 0.9078 0.9464 0.928 0.871 0.9767 1.4204
026_8061_00290830 Brain 8061 GB-1 456 3.686496 0.613 0.692 0.809
0.8255 0.8977 0.877 0.9922 0.9416 0.8923 026_374_00283180 Brain 374
U-251 MG 456 3.933399 0.654 0.733 0.861 0.8164 0.8981 0.891 0.9222
0.9976 1.1342 026_343_00283160 Brain 343 PFSK-1 456 3.963833 0.626
0.904 1.007 1.0229 0.934 1.078 1.0411 1.0084 1.052 026_393_00283190
Brain 393 YKG-1 456 3.987729 0.646 0.714 0.77 0.8178 0.8374 0.848
0.953 0.8805 1.0209 026_8028_00287630 Brain 8028 D-263MG 456
4.157483 0.662 0.709 0.734 0.7813 0.8341 0.839 0.9023 0.9333 1.0435
026_379_00283140 Brain 379 GAMG 456 4.214093 0.681 0.701 0.829
0.8043 0.8412 0.83 0.9322 0.8908 1.046 026_8019_00293320 Brain 8019
CAS-1 456 4.309856 0.732 0.772 0.825 0.9222 0.9783 1.227 0.8865
0.8407 1.1628 026_8001_00285100 Brain 8001 8-MG-BA 456 4.345495
0.688 0.845 0.88 0.8958 0.9285 0.912 0.9148 0.9398 1.07
026_351_00283450 Brain 351 LN-18 456 4.481815 0.685 0.822 0.862
0.8746 0.8966 0.902 0.9022 0.9311 0.9325 026_357_00283410 Brain 357
H4 456 4.481909 0.721 0.808 0.876 0.858 0.8822 0.94 1.0232 0.9287
1.0614 026_8085_00293731 Brain 8085 KINGS-1 456 4.48888 0.797 0.781
0.82 0.9645 0.9641 1.015 0.9981 1.0442 0.974 026_350_00284910 Brain
350 M059J 456 4.537622 0.723 0.834 0.869 0.8888 0.9473 0.909 0.9144
1.0191 1.0818 026_8015_00308070 Brain 8015 Becker 456 4.548091
0.696 0.814 0.876 0.8121 0.825 0.889 0.917 0.9113 1.0138
026_8160_00287680 Brain 8160 SF539 456 4.738405 0.726 0.919 0.827
0.8875 0.9392 0.934 0.952 1.0525 0.9285 026_8159_00287670 Brain
8159 SF268 456 4.804304 0.786 0.825 0.923 0.9225 0.9772 0.99 0.978
0.9754 0.9699 026_359_00283100 Brain 359 Daoy 456 4.81575 0.683
0.74 0.796 0.8102 0.876 0.896 0.9135 0.9254 0.9257
026_8217_00290890 Brain 8217 SK-MG-1 456 4.83361 0.725 0.784 0.83
0.844 0.8849 0.901 0.9119 0.9353 0.9859 026_342_00285160 Brain 342
SW 1783 456 4.84471 0.796 0.905 0.896 0.9349 1.0662 0.935 1.0347
1.0148 1.08 026_8029_00288240 Brain 8029 D-336MG 456 4.925588 0.792
0.892 0.997 0.8872 0.9556 0.944 1.105 1.0724 1.1057
026_8030_00295500 Brain 8030 D-392MG 456 4.966575 0.791 0.866 0.824
0.9331 0.8638 1.022 0.9531 0.9688 1.0531 026_8089_00287440 Brain
8089 KNS-81-FD 456 5.077406 0.804 0.89 0.869 0.8138 0.8421 1.049
1.0747 0.9317 1.1222 026_8138_00285290 Brain 8138 NMC-G1 456
5.086457 0.771 0.817 0.763 0.762 0.8138 0.887 0.968 1.0104 1.049
026_8139_00285130 Brain 8139 no-10 456 5.147267 0.824 0.888 0.928
0.9069 0.9436 1.031 0.943 1.05 1.0543 026_8083_00293720 Brain 8083
KALS-1 456 5.203248 0.817 0.765 0.902 0.9843 0.9376 0.915 0.9382
1.0116 0.9496 026_378_00284880 Brain 378 DK-MG 456 5.2953 0.789
0.757 0.713 0.7179 0.7297 0.849 0.8425 0.9101 1.0114
026_383_00284900 Brain 383 LN-405 456 5.313289 0.839 0.886 0.901
0.937 0.8868 1.041 0.949 1.0816 1.0986 026_8032_00293340 Brain 8032
D-542MG 456 5.342096 0.822 0.856 0.927 0.9108 0.8977 0.934 0.9191
0.9743 0.9895 026_344_00282720 Brain 344 LNZTA3WT4 456 5.43481
0.777 1.136 0.931 0.9386 0.9371 1.018 1.0441 1.0001 0.9944
026_8167_00290910 Brain 8167 SNB75 456 5.474524 0.76 0.772 0.83
0.8023 0.8147 1.01 0.8804 1.0523 0.9382 026_8087_00285270 Brain
8087 KNS-42 456 5.484622 0.862 0.858 0.846 0.8777 0.9301 0.972
0.9923 1.0701 1.0135 026_354_00287481 Brain 354 U-87 MG 456
5.588679 0.851 0.949 0.942 0.9605 0.9939 1.004 1.0222 0.9964 1.0855
026_8140_00285300 Brain 8140 no-11 456 5.608459 0.844 0.885 0.896
0.959 0.9411 0.893 0.9627 1.0352 0.9432 026_8221_00284860 Brain
8221 D-423MG 456 5.731372 0.807 0.783 0.798 0.8469 0.8833 0.945
0.9877 0.9131 0.9215 026_348_00283400 Brain 348 DBTRG-05MG 456
5.749405 0.792 0.75 0.763 0.7919 0.7917 0.856 0.9393 0.9013 0.9922
026_341_00285310 Brain 341 SW 1088 456 5.805148 0.869 0.907 0.895
0.8893 0.9105 0.919 0.966 0.8961 1.0957 026_356_00283420 Brain 356
Hs 683 456 5.858982 0.841 0.993 0.888 0.8789 0.8908 1.04 0.8743
0.906 1.058 026_8031_00287640 Brain 8031 D-502MG 456 5.99897 0.825
0.819 0.771 0.7649 0.8658 0.834 0.9457 0.9922 0.9572
026_8224_00284870 Brain 8224 D-566MG 456 6.026403 0.839 0.84 0.858
0.8841 0.9252 0.941 0.968 0.9815 0.9816 026_389_00284920 Brain 389
MOG-G-UVW 456 6.074777 0.861 0.876 0.879 0.9129 0.9004 0.902 1.0597
0.9108 1.0304 026_341_00283470 Brain 341 SW 1088 456 6.105387 0.913
0.89 0.897 0.897 0.9463 0.931 0.9486 0.9506 1.1024 026_375_00284850
Brain 375 42-MG-BA 456 6.106423 0.896 0.892 0.932 0.944 0.9633
0.976 0.9735 0.9871 1.0285 026_1122_00283170 Brain 1122 SF-295 456
6.112956 0.879 0.909 0.909 0.9301 0.9215 0.946 0.9199 0.9433 1.0329
026_8158_00290650 Brain 8158 SF126 456 6.158755 0.86 0.97 1 1.0676
1.0025 1.187 0.9851 0.9627 1.1324 026_340_00285250 Brain 340
CCF-STTG1 456 6.170298 0.851 0.911 0.916 0.9011 0.9748 0.919 0.9109
0.9323 0.9837 026_380_00284890 Brain 380 GMS-10 456 6.23472 0.842
0.885 0.853 0.8466 0.8914 0.914 0.8882 1.0019 1.0545
026_354_00290361 Brain 354 U-87 MG 456 6.315816 0.93 0.874 0.977
0.9346 0.9336 1.084 0.9295 1.062 1.073 026_8063_00290841 Brain 8063
GI-1 456 6.443002 0.809 0.877 0.913 0.9018 0.9184 0.86 0.9241
0.9072 0.9709 026_8027_00293330 Brain 8027 D-247MG 456 6.453915
0.928 0.905 0.908 0.8398 0.8592 0.911 0.9463 0.9529 0.9044
026_346_00283390 Brain 346 A172 456 6.640511 0.941 0.953 0.995
1.0208 0.9242 1.026 1.0141 1.0236 1.0356 026_8089_00291170 Brain
8089 KNS-81-FD 456 7.067458 0.854 0.823 0.854 0.8437 0.8524 0.889
0.8917 0.9075 0.9111 026_355_00285180 Brain 355 U-118 MG 456
7.408088 0.965 1.005 0.945 0.9352 0.9307 0.932 0.9407 0.945 0.9851
026_347_00282740 Brain 347 T98G 456 7.802123 0.986 1.098 0.951
1.0602 0.9957 1.038 1.0462 0.9753 0.982 026_417_00271110 Breast 417
DU4475 456 -3.0044 0.119 0.118 0.124 0.1202 0.1235 0.121 0.164
0.4789 0.6922 26_465_00271670 Breast 465 MRK-nu-1 456 1.744742
0.115 0.398 0.572 0.6893 0.7807 0.868 0.921 0.9463 1.1175
026_438_00273540 Breast 438 HCC1599 456 1.968855 0.352 0.459 0.617
0.7517 0.8021 0.899 0.9102 0.9757 1.0216 026_435_00271290 Breast
435 HCC1187 456 2.132259 0.375 0.482 0.801 0.7466 0.7917 0.794
0.9164 0.9352 0.9707 026_403_00271400 Breast 403 MCF7 456 2.857648
0.493 0.634 0.765 0.8761 0.9706 0.973 1.0249 1.0247 1.0456
026_401_00273450 Breast 401 MDA-MB-468 456 3.05753 0.415 0.718
0.931 0.9526 0.909 0.954 0.9512 1.0282 0.9601 26_451_00271640
Breast 451 CAL-85-1 456 3.06898 0.489 0.695 0.808 0.7646 0.8147
0.885 0.8459 1.0885 0.9906 026_404_00273430 Breast 404 MDA-MB-231
456 3.086092 0.483 0.731 0.783 0.824 0.8358 0.874 0.9446 0.9609
1.0599 026_418_00271550 Breast 418 Hs 578T 456 3.125956 0.071 0.614
0.832 0.7916 0.8611 0.836 0.9569 0.9556 1.0186 026_402_00272120
Breast 402 CAMA-1 456 3.166443 0.55 0.636 0.773 0.8152 0.9017 0.886
0.906 0.9417 0.9695 026_426_00274200 Breast 426 HCC1569 456
3.228782 0.571 0.713 0.945 0.9056 0.9414 0.938 0.9814 1.028 1.0046
026_431_00271130 Breast 431 HCC1806 456 3.337319 0.566 0.672 0.68
0.6852 0.6987 0.765 0.8378 1.0081 1.1468 026_414_00271900 Breast
414 AU565 456 3.409308 0.533 0.816 0.949 0.9831 0.9841 0.924 0.9729
0.9876 1.1475 026_452_00272130 Breast 452 COLO-824 456 3.645131
0.358 0.743 0.843 0.9519 0.9643 0.904 0.9021 1.0288 0.9665
026_416_00271360 Breast 416 BT-549 456 3.723874 0.659 0.818 0.911
0.9492 0.9446 0.965 0.9707 0.9785 1.0314 026_8144_00274240 Breast
8144 OCUB-M 456 3.727884 0.44 0.801 0.89 0.9127 1.0139 1.046 0.9729
0.96 1.0167 026_432_00271960 Breast 432 HCC70 456 3.73179 0.605
0.733 0.855 0.8647 0.8214 0.887 0.9582 0.9394 1.1987
026_457_00273420 Breast 457 EVSA-T 456 3.968951 0.686 0.879 0.927
0.9385 0.9505 0.975 0.9713 0.9606 1.0214 026_466_00274370 Breast
466 YMB-1-E 456 3.997753 0.654 0.836 0.87 0.9113 0.9082 0.908
0.9623 1.0457 0.9738 026_441_00285120 Breast 441 HCC2157 456
3.997874 0.474 1.044 0.849 0.8244 0.6122 0.948 1.0739 0.6695
1.0405
026_443_00271990 Breast 443 MDA-MB-330 456 4.004084 0.601 0.92 0.94
0.9085 0.9663 0.971 0.9817 0.9257 1.0607 026_436_00271300 Breast
436 HCC1395 456 4.036641 0.703 0.897 0.946 0.9395 1.0136 1.033
1.0207 1.0407 1.1296 026_412_00277190 Breast 412 UACC-893 456
4.234383 0.421 0.859 0.854 0.8448 0.9188 0.999 0.9512 1.0422 0.9971
026_450_00271390 Breast 450 CAL-51 456 4.319545 0.678 0.725 0.783
0.8043 0.8395 0.899 0.9313 0.9642 1.0975 026_449_00271380 Breast
449 CAL-148 456 4.389344 0.747 0.676 1.08 0.9571 0.7093 0.888 0.981
0.8213 1.2196 026_434_00271920 Breast 434 HCC1143 456 4.464516
0.726 0.808 0.808 0.822 0.8993 0.923 0.919 0.949 1.1427
026_433_00276270 Breast 433 HCC202 456 4.571252 0.686 0.9 0.944
0.8549 0.8115 0.943 0.972 0.9241 0.9618 026_422_00274230 Breast 422
MDA-MB-175-VII 456 4.594595 0.7 0.73 0.74 0.7666 0.8545 0.848
0.9155 0.9541 0.9487 026_461_00272170 Breast 461 MFM-223 456
4.656681 0.787 0.875 0.908 0.9465 0.988 1.047 1.0015 1.1431 1.0159
026_427_00271330 Breast 427 MDA-MB-453 456 4.669025 0.795 0.819
0.921 0.9998 0.9757 0.998 1.0349 1.0264 1.1651 026_448_00271370
Breast 448 CAL-120 456 4.779687 0.744 0.746 0.796 0.8032 0.8728
0.85 0.9852 1.0071 1.0287 026_411_00271420 Breast 411 UACC-812 456
5.072094 0.82 0.894 0.873 0.9564 0.9958 0.944 0.9931 1.0326 0.9659
026_442_00273480 Breast 442 HCC2218 456 5.225292 0.614 0.923 0.963
1.0597 1.042 1.043 1.0714 1.0498 1.0318 026_398_00272150 Breast 398
HCC1428 456 5.256241 0.85 0.899 1.013 0.8915 0.9851 0.983 0.9959
0.9937 1.0331 026_464_00308490 Breast 464 T47D 456 5.26439 0.825
0.853 0.869 0.8588 0.823 0.978 0.9397 0.9562 0.968 026_400_00273440
Breast 400 MDA-MB-436 456 5.286367 0.725 0.844 0.848 0.8326 0.8721
0.851 0.8881 0.9805 0.933 026_437_00280161 Breast 437 HCC1500 456
5.288806 0.796 0.982 0.913 0.8847 0.9397 1.088 0.8523 0.9109 0.9883
026_440_00271950 Breast 440 HCC1954 456 5.303093 0.807 0.813 0.885
0.8155 0.8463 0.861 1.0199 1.0209 0.9481 026_413_00271930 Breast
413 HCC1419 456 5.337084 0.839 0.926 0.932 0.9634 0.9633 0.941
0.9595 0.9516 1.1518 026_410_00272180 Breast 410 ZR-75-30 456
5.373081 0.83 0.857 0.88 0.8529 0.9183 0.913 0.911 1.03 0.9685
026_439_00271940 Breast 439 HCC1937 456 5.44243 0.841 0.952 0.975
1.0253 0.9632 0.953 1.0302 1.0262 1.0774 026_408_00271350 Breast
408 BT-20 456 5.735872 0.871 0.955 0.93 0.9913 0.9764 0.972 1.0674
0.9818 1.1818 026_399_00272160 Breast 399 MDA-MB-415 456 5.879088
0.884 0.875 0.885 0.8851 0.9175 0.914 0.9179 0.9049 0.9735
026_458_00274220 Breast 458 HDQ-P1 456 6.259493 0.84 1.072 1.136
1.0763 1.0797 1.042 1.0437 1.0231 1.0073 026_397_00274350 Breast
397 HCC38 456 6.744219 0.874 1.053 1.023 1.0222 0.9856 0.93 0.9274
0.9402 0.9941 026_405_00274360 Breast 405 MDA-MB-361 456 6.792889
0.935 0.959 0.987 0.9819 0.9119 0.921 0.9597 0.998 0.9922
026_425_00280231 Breast 425 MDA-MB-157 456 7.000504 0.906 1.069
1.05 0.9597 0.9229 0.966 0.9422 0.9712 0.9545 026_454_00272140
Breast 454 EFM-192A 456 7.097342 0.942 1.032 1.041 1.0545 0.9812
0.979 1.0186 1.0191 0.9742 026_420_00271541 Breast 420 BT-474 456
7.458914 0.191 1.089 1.053 1.0824 1.0008 0.955 0.9764 0.9678 1.0149
026_453_00273410 Breast 453 EFM-19 456 8.207256 1.096 1.286 1.208
1.1112 1.0544 1.042 1.0064 1.0474 1.0572 026_415_00316440 Breast
415 BT-483 456 8.21654 1.148 1.275 1.181 1.1462 1.089 1.147 1.0671
1.0911 1.1123 026_8176_00316650 Cervix 8176 TC-YIK 456 0.842618
0.29 0.416 0.488 0.5211 0.5993 0.669 0.7672 0.7877 1.0572
026_479_00264920 Cervix 479 HT-3 456 1.420025 0.257 0.608 0.574
0.5844 0.5901 0.637 0.8649 0.8189 0.8617 026_478_00271910 Cervix
478 C-33 A 456 2.72591 0.472 0.615 0.658 0.6776 0.6753 0.734 0.7964
0.8383 0.9143 026_478_00269410 Cervix 478 C-33 A 456 2.976483 0.445
0.701 0.714 0.731 0.8172 0.771 0.9038 0.852 0.9634 026_493_00268830
Cervix 493 ME-180 456 3.07379 0.509 0.667 0.695 0.7355 0.6828 0.732
0.7953 0.9097 1.0456 026_476_00269050 Cervix 476 C-4 I 456 3.232632
0.508 0.792 0.857 0.8645 0.8671 0.933 1.0078 1.0484 1.1636
026_8145_00271140 Cervix 8145 OMC-1 456 3.295968 0.531 0.786 0.84
0.8546 0.9307 0.911 1.0261 1.0441 1.0392 026_484_00263710 Cervix
484 Ca Ski 456 3.639931 0.327 0.715 0.879 0.8811 0.8284 0.817
0.9599 1.0361 1.165 026_469_00264610 Cervix 469 HeLa 456 3.981495
0.311 0.783 0.933 0.9784 0.9807 0.982 0.9321 0.9429 0.9946
026_493_00262480 Cervix 493 ME-180 456 4.007923 0.639 0.799 0.885
0.9474 0.9337 0.873 0.9457 0.8694 0.8949 026_474_00269100 Cervix
474 SiHa 456 4.596545 0.742 0.846 0.832 0.8807 0.9276 0.898 1.0004
0.9967 1.1646 026_482_00262520 Cervix 482 SISO 456 5.313141 0.851
0.946 0.96 0.9752 0.9888 0.276 0.975 0.947 0.9669 026_482_00274250
Cervix 482 SISO 456 5.375837 0.842 0.881 0.887 0.9541 0.9605 0.96
0.9679 1.044 1.0481 026_482_00269740 Cervix 482 SISO 456 5.709731
0.838 0.89 0.925 0.9202 0.9445 0.931 0.9609 0.9695 0.9938
026_468_00264600 Cervix 468 DoTc2 4510 456 5.749229 0.363 0.92
0.961 0.9796 0.9907 0.984 1.01 0.9525 0.9858 026_473_00264650
Cervix 473 SW756 456 5.953892 0.162 0.868 0.813 0.7938 0.8104 0.779
0.9163 0.8951 0.9568 026_491_00264830 Cervix 491 5KG-IIIa 456
6.261878 0.721 1.002 1.008 1.007 1.0125 0.97 1.0049 0.9721 0.9931
026_476_00264900 Cervix 476 C-4 I 456 6.792994 0.225 0.991 0.994
0.9083 0.8797 0.89 0.982 0.9123 0.9144 026_474_00264930 Cervix 474
SiHa 456 7.505839 0.532 0.824 0.865 0.9092 0.9029 0.879 0.8904
0.8746 0.8785 026_472_00264630 Cervix 472 MS751 456 7.679336 1.023
0.945 0.949 0.9806 0.9744 0.979 0.9915 0.9775 0.9824
026_8180_00276230 Esophagus 8180 TE-15 456 0.823778 0.313 0.365
0.448 0.4963 0.618 0.64 0.8902 0.6859 1.0361 026_502_00276550
Esophagus 502 KYSE-450 456 1.219587 0.422 0.41 0.448 0.5484 0.5334
0.703 0.7332 0.8892 0.853 026_497_00274050 Esophagus 497 KYSE-150
456 1.304746 0.328 0.402 0.467 0.567 0.6585 0.801 0.8957 0.9976
0.9719 026_8252_00276570 Esophagus 8252 OACp4C 456 1.678468 0.101
0.3 0.567 0.6503 0.7826 0.861 0.8531 0.9243 0.9446 026_496_00276530
Esophagus 496 KYSE-140 456 2.209734 0.522 0.516 0.476 0.5824 0.6741
0.815 0.8515 0.806 0.8896 026_8233_00278570 Esophagus 8233 ESO26
456 2.595405 0.493 0.568 0.679 0.7293 0.7791 0.852 0.8801 1.0449
0.8251 026_8184_00282680 Esophagus 8184 TE-6 456 2.946928 1.152
0.677 0.704 0.6755 0.7901 0.89 1.1989 1.0724 0.9825
026_8277_00276670 Esophagus 8277 TE-4 456 3.072573 0.518 0.605
0.768 0.8018 0.8549 0.883 0.9093 0.8952 0.9579 026_506_00277170
Esophagus 506 OE19 456 3.143883 0.544 0.752 0.743 0.8799 1.0437
0.937 1.0332 1.0004 1.0701 026_8179_00276220 Esophagus 8179 TE-12
456 3.163617 0.183 0.742 0.633 0.6476 0.7356 0.839 0.7964 0.8189
0.8318 026_8185_00276250 Esophagus 8185 TE-8 456 3.50176 0.201
0.606 0.658 0.7374 0.6587 0.744 0.775 0.681 0.9258
026_8184_00293680 Esophagus 8184 TE-6 456 3.545508 0.552 0.767
0.935 0.8745 0.8626 0.742 1.0239 0.863 0.9232 026_8178_00280260
Esophagus 8178 TE-10 456 3.804003 0.664 0.735 0.78 0.9124 0.8781
1.01 1.0331 0.9777 1.0127 026_499_00276630 Esophagus 499 KYSE-270
456 4.027681 0.678 0.727 0.767 0.7928 0.8774 0.984 0.9911 1.0386
1.0007 026_509_00276620 Esophagus 509 KYSE-220 456 4.075564 0.637
0.656 0.682 0.7368 0.7969 0.844 0.873 0.9181 0.9229
026_8235_00276520 Esophagus 8235 FLO-1 456 4.100409 0.604 0.732
0.778 0.7882 0.8071 0.864 0.8729 0.8842 0.9483 026_8251_00276640
Esophagus 8251 OACM5-1 456 4.181038 0.673 0.784 0.878 0.8883 0.9366
0.926 0.9214 0.9309 0.9714 026_8186_00282690 Esophagus 8186 TE-9
456 4.291684 0.689 0.795 0.742 0.8146 0.856 0.846 1.0123 0.8588
1.0654 026_495_00274190 Esophagus 495 COLO-680N 456 4.332573 0.742
0.818 0.865 0.9122 0.97 0.96 1.1262 0.9684 1.0444 026_510_00273560
Esophagus 510 KYSE-50 456 4.431288 0.658 0.743 0.781 0.8044 0.8395
0.815 0.8665 0.9009 1.0802 026_8186_00292740 Esophagus 8186 TE-9
456 4.438695 0.739 0.769 0.893 0.8667 0.8573 0.944 1.0623 1.0025
1.1458 026_503_00274070 Esophagus 503 KYSE-510 456 4.484808 0.731
0.796 0.816 0.8942 0.9487 0.912 0.9785 1.0104 1.0221
026_504_00276560 Esophagus 504 KYSE-520 456 4.773857 0.716 0.731
0.748 0.7768 0.8124 0.867 0.9459 0.995 0.9576 026_8208_00276600
Esophagus 8208 HCE-4 456 4.88732 0.748 0.785 0.808 0.8645 0.9033
0.954 0.9197 0.9231 0.9878 026_512_00274080 Esophagus 512 T.T 456
4.943912 0.781 0.839 0.89 0.9002 0.9769 0.929 0.9845 0.9744 1.0152
026_8268_00276650 Esophagus 8268 SK-GT-4 456 5.079273 0.835 0.788
0.758 0.735 0.7503 0.833 0.9063 0.9163 0.972 026_505_00274210
Esophagus 505 KYSE-70 456 5.137973 0.794 0.857 0.892 0.8943 0.9411
0.957 0.9493 0.9907 1.0431 026_508_00278520 Esophagus 508 OE33 456
5.424479 0.701 0.81 0.783 0.7873 0.7763 0.833 0.8596 0.903 0.9287
026_498_00276540 Esophagus 498 KYSE-180 456 5.533652 0.84 0.853
0.898 0.9109 0.9353 0.924 0.9336 0.9199 0.942 026_8202_00276660
Esophagus 8202 TE-11 456 5.740526 0.822 0.845 0.848 0.8684 0.8821
0.913 0.9297 0.9627 1.0605 026_8039_00276580 Esophagus 8039
EC-GI-10 456 5.844841 0.878 0.905 0.892 0.951 0.9277 0.926 0.9555
1.04 1.0068 026_501_00274060 Esophagus 501 KYSE-410 456 6.135532
0.762 0.836 0.846 0.8398 0.8691 0.887 0.9278 0.9729 0.9854
026_8246_00276610 Esophagus 8246 KYAE-1 456 6.1525 0.887 0.93 0.92
0.9326 0.9185 0.978 0.97 0.9663 0.9315 026_507_00278510 Esophagus
507 OE21 456 6.23551 0.922 0.838 0.875 0.7771 0.8079 0.985 0.9753
0.9667 0.9919 026_8183_00276240 Esophagus 8183 TE-5 456 6.777236
0.111 1.028 0.936 1.0341 1.0898 0.876 0.8843 0.9699 0.8561
026_8177_00282670 Esophagus 8177 TE-1 456 7.848711 1.022 1.1 0.912
1.0973 1.0839 1.065 1.0874 1.0775 1.0749 026_545_00260020 Head
& Neck 545 DOK 456 -0.22061 0.308 0.451 0.33 0.4014 0.4436
0.534 0.6634 0.7589 0.8693 026_1217_00255750 Head & Neck 1217
H3118 456 0.314816 0.254 0.413 0.453 0.4692 0.5199 0.565 0.691
0.6788 0.7738 026_526_00308740 Head & Neck 526 PCI-4B 456
1.887412 0.418 0.409 0.489 0.7595 0.7429 0.873 0.9067 0.918 1.0403
026_530_00260620 Head & Neck 530 PCI-30 456 1.98819 0.142 0.356
0.628 0.7119 0.7851 0.825 0.9139 0.8747 0.8098 026_552_00252890
Head & Neck 552 SAT 456 2.534279 0.382 0.613 0.727 0.8554
0.7778 0.977 0.8474 0.7957 0.9605 026_550_00258980 Head & Neck
550 SCC-4 456 2.544916 0.464 0.564 0.904 0.7519 0.7827 0.709 0.7715
0.8596 0.9574 026_1224_00256200 Head & Neck 1224 SCC-9 456
2.68588 0.466 0.633 0.641 0.6635 0.6231 0.758 0.8287 0.8854 0.9263
026_1223_00259190 Head & Neck 1223 SCC-25 456 2.934793 0.193
0.664 0.718 0.8957 0.937 0.932 1.0111 0.988 1.0055 026_548_00261030
Head & Neck 548 RPMI 2650 456 3.095229 0.489 0.696 0.761 0.8664
0.8685 0.911 0.9177 0.9218 0.9624 026_517_00308680 Head & Neck
517 JHU-011 456 3.112112 0.536 0.616 0.696 0.7845 0.8433 0.813
0.9157 0.9495 1.0827 026_8011_00257080 Head & Neck 8011
BB30-HNC 456 3.217966 0.541 0.677 0.734 0.7445 0.7909 0.917 0.9622
0.9832 1.0203 026_553_00259140 Head & Neck 553 OSC-20 456
3.378306 0.587 0.751 0.792 0.8761 0.9722 0.983 1.0097 0.974 0.999
026_556_00257220 Head & Neck 556 SKN-3 456 3.392608 0.207 0.626
0.805 0.7174 0.6353 0.718 0.757 0.8593 0.8383 026_536_00256080 Head
& Neck 536 BHY 456 3.443799 0.593 0.821 0.877 0.8931 0.9405
1.049 1.0289 0.9991 1.0281 026_561_00257110 Head & Neck 561
Ca9-22 456 3.710495 0.634 0.867 0.849 0.916 0.9493 0.99 1.0155
1.0225 1.0222 026_532_00308750 Head & Neck 532 PCI-6A 456
3.742916 0.628 0.684 0.811 0.8452 0.8862 0.945 0.9298 0.9056 0.9974
026_8012_00266550 Head & Neck 8012 BB49-HNC 456 3.77782 0.46
0.775 0.9 0.8967 0.924 0.968 0.9717 1.0116 0.9946 026_8100_00256170
Head & Neck 8100 LB771 -HNC 456 3.926882 0.701 0.899 0.929
0.9987 0.9867 0.989 1.0145 0.9621 0.9588 026_1222_00253030 Head
& Neck 1222 SCC-15 456 4.189701 0.411 0.772 0.855 0.8291 0.7747
1.02 0.8664 0.932 0.9717 026_533_00260900 Head & Neck 533
PCI-15A 456 4.236361 1.325 1.15 0.832 0.8691 0.8188 0.863 0.9549
1.0427 1.0471 26_547_00314070 Head & Neck 547 KOSC-2 cl3-43 456
4.563275 0.708 0.872 0.897 0.9 0.8378 0.941 1.0131 0.9825 0.9292
026_544_00256140 Head & Neck 544 Detroit 562 456 4.601961 0.746
0.964 0.951 0.9801 0.9656 0.961 0.9765 0.9793 0.9061
026_543_00256100 Head & Neck 543 BICR 78 456 4.851894 0.728
0.818 0.868 0.8349 0.9283 0.925 0.9696 0.966 0.9649
026_537_00256120 Head & Neck 537 CAL-33 456 5.580216 0.834
0.892 0.921 0.9097 0.9511 0.935 0.9977 1.0062 0.9837
026_549_00256220 Head & Neck 549 HO-1-N-1 456 5.63511 0.866
0.873 0.883 0.8726 0.8907 0.97 0.987 0.9898 1.0714 026_557_00259180
Head & Neck 557 SAS 456 6.035072 0.913 0.97 0.967 0.9738 0.9866
1.043 1.0896 1.0407 1.0277 026_534_00256110 Head & Neck 534 CAL
27 456 6.045151 0.85 0.904 0.912 0.9312 0.925 0.939 0.9758 0.9901
0.9927 026_542_00269190 Head & Neck 542 BICR 31 456 6.045414
0.903 0.908 0.952 0.9408 0.977 0.97 0.9966 1.0002 1.0269
026_530_00262500 Head & Neck 530 PCI-30 456 6.119674 0.884
0.955 1.145 0.9682 1.009 0.988 0.9261 1.0121 0.9861
026_540_00258490 Head & Neck 540 BICR 10 456 6.128463 0.839
1.048 1.049 1.0556 1.0464 1.041 1.0301 0.974 0.9953
026_541_00256090 Head & Neck 541 BICR 22 456 6.156446 0.828
1.076 1.058 0.9602 1.0898 1.064 1.0874 0.9689 1.0462
026_8003_00263440 Head & Neck 8003 A253 456 6.291179 0.877
0.857 0.859 0.8594 0.9753 0.982 0.9985 0.9367 0.969
026_535_00256160 Head & Neck 535 FaDu 456 6.303565 0.841 1.063
1.091 1.0795 1.0793 1.034 1.0404 1.0191 1.0095 026_554_00257210
Head & Neck 554 OSC-19 456 6.322489 0.938 0.942 0.914 0.9247
0.9682 1.004 1.0285 1.0504 1.0506 026_559_00256250 Head & Neck
559 HSC-3 456 6.797258 0.909 1.024 1.057 1.0632 1.0572 1.029 1.0059
0.9701 0.9782 026_8071_00256760 Head & Neck 8071 HCE-T 456
6.917353 0.576 1.146 1.274 1.3514 1.1688 1.173 1.0336 1.1999 0.8849
026_521_00257180 Head & Neck 521 JHU-022 456 7.049785 0.896
0.846 0.901 0.8757 0.9278 0.886 0.9103 0.9436 0.9699
026_555_00256270 Head & Neck 555 KON 456 7.114386 0.946 0.982
0.977 0.9826 0.9957 0.96 0.9991 0.961 0.9207 026_538_00258510 Head
& Neck 538 HN 456 7.118392 0.947 1.007 0.984 1.0014
1.0052 0.998 1.0105 0.9648 0.9952 026_546_00259150 Head & Neck
546 PE/CA-PJ15 456 7.26219 0.96 0.963 0.931 0.9179 0.9698 0.969
0.9884 1.006 0.9974 026_560_00256260 Head & Neck 560 HSC-4 456
7.41962 1.005 0.994 1.003 0.9486 1.2114 0.897 0.9756 0.9433 0.9255
026_551_00259130 Head & Neck 551 HO-1-u-1 456 7.903602 0.999
1.037 1.039 1.0722 1.0616 1.067 1.0655 1.0038 0.9405
026_558_00256240 Head & Neck 558 HSC-2 456 8.185707 1.148 1.077
0.985 1.1269 1.0034 1.093 0.9608 0.9252 0.9888 026_531_00258970
Head & Neck 531 PCI-38 456 8.493869 1.046 1.067 1.063 1.0751
1.0712 0.965 1.0236 1.0421 0.9848 026_570_00293670 Intestine 570
SK-CO-1 456 -0.30187 0.268 0.258 0.267 0.2555 0.3743 0.418 0.7306
0.9824 0.9276 026_8153_00295901 Intestine 8153 RKO 456 0.044041
0.222 0.197 0.246 0.3161 0.4612 0.693 0.7818 0.9061 1.1131 18
Intestine 586 COLO 205 456 0.350012 0.083 0.182 0.268 0.3978 0.5206
0.706 0.8518 0.9472 0.9745 026_582_00295550 Intestine 582 LoVo 456
0.399717 0.325 0.396 0.385 0.4168 0.4097 0.533 0.75 0.8239 0.9276
026_8108_00298530 Intestine 8108 LS-513 456 0.423432 0.094 0.146
0.195 0.2527 0.5673 0.876 1.1755 1.0179 1.1926 026_8274_00258540
Intestine 8274 SNU-61 456 0.467862 0.432 0.599 0.539 0.5369 0.5735
0.603 0.7283 0.7462 0.9243 026_8136_00260060 Intestine 8136
NCI-H747 456 0.578701 0.356 0.465 0.488 0.5338 0.5471 0.613 0.7238
0.8107 0.9473 026_574_00298390 Intestine 574 CL-11 456 0.894127
0.381 0.401 0.395 0.4593 0.6002 0.69 0.749 0.8474 0.9789
026_589_00295371 Intestine 589 HCT 116 456 0.931657 0.336 0.381
0.435 0.4732 0.5791 0.862 1.1049 1.0627 1.1072 026_608_00293620
Intestine 608 CCK-81 456 1.046144 0.242 0.394 0.399 0.4912 0.6325
0.738 0.9653 0.9069 0.9123 026_610_00293660 Intestine 610 RCM-1 456
1.475794 0.45 0.416 0.414 0.6252 0.6775 0.804 0.981 1.0277 1.0565
026_569_00295390 Intestine 569 HT-29 456 1.526342 0.48 0.464 0.5
0.5547 0.5893 0.74 0.9536 0.9048 0.9241 026_8271_00314300 Intestine
8271 SNU-175 456 1.739925 0.382 0.522 0.535 0.5651 0.6481 0.701
0.7957 0.9458 0.9851 026_8108_00296610 Intestine 8108 LS-513 456
1.815428 0.37 0.448 0.511 0.6156 0.8074 0.933 0.9905 0.9579 0.9991
026_592_00295380 Intestine 592 HT115 456 1.869689 0.377 0.436 0.5
0.6258 0.8213 1.101 1.0858 1.0169 1.1148 026_606_00293630 Intestine
606 HCC-56 456 1.929923 0.646 0.514 0.536 0.5915 0.7207 0.867
0.8829 0.9131 0.9417 026_8107_00296450 Intestine 8107 LS-411N 456
2.297731 0.497 0.55 0.585 0.5585 0.6884 0.755 0.8626 0.8921 0.9253
026_595_00295420 Intestine 595 LS180 456 2.32965 0.453 0.515 0.551
0.6999 0.8666 0.91 1.0102 0.9178 1.1155 026_8169_00295910 Intestine
8169 SNU-C2B 456 2.617884 0.503 0.566 0.598 0.7373 0.671 0.774
0.9066 0.8796 0.9886 026_564_00292860 Intestine 564 NCI-H630 456
2.627492 0.496 0.556 0.55 0.7713 1.0007 0.964 0.9563 0.8783 0.9169
026_603_00292731 Intestine 603 SW837 456 2.797981 0.525 0.597 0.586
0.7811 0.7763 1.002 1.023 1.0218 1.0696 026_588_00295360 Intestine
588 GP5d 456 2.925873 0.521 0.65 0.67 0.679 0.8325 0.964 1.0479
1.1013 1.0678 026_598_00302650 Intestine 598 SW 1417 456 3.077195
0.616 0.696 0.742 0.6531 1.034 1.029 1.0798 0.8321 1.2207
026_8276_00296000 Intestine 8276 SNU-C5 456 3.153347 0.603 0.592
0.612 0.6327 0.7283 0.812 0.8824 0.9411 0.9768 026_593_00295530
Intestine 593 HT55 456 3.164186 0.589 0.646 0.61 0.7857 0.7974 1.03
1.1134 1.0941 1.1175 026_8106_00264670 Intestine 8106 LS-123 456
3.191118 0.547 0.687 0.783 0.7554 0.8823 0.879 0.9202 0.965 0.9375
026_599_00296340 Intestine 599 SW 1463 456 3.237942 0.572 0.642
0.631 0.6255 0.6275 0.757 0.9237 0.9401 0.9681 026_8086_00296431
Intestine 8086 KM12 456 3.240654 0.564 0.657 0.65 0.6739 0.8062
0.903 0.8847 0.9825 0.9793 026_587_00302320 Intestine 587 COLO 741
456 3.269221 0.615 0.622 0.694 0.6624 0.7236 0.816 1.1047 1.0995
1.0783 026_8273_00295970 Intestine 8273 SNU-407 456 3.390403 0.577
0.622 0.663 0.7325 0.7399 0.745 0.8059 0.8644 1.0342
026_8270_00304630 Intestine 8270 SNU-1040 456 3.643993 0.604 0.78
0.799 0.8256 0.8028 0.92 0.9246 0.9919 1.2066 026_8168_00295990
Intestine 8168 SNU-C1 456 3.717495 0.645 0.673 0.685 0.6875 0.8368
0.739 0.8513 1.1115 1.0883 026_8275_00256210 Intestine 8275 SNU-81
456 3.903109 0.338 0.659 0.68 0.6942 0.7065 0.761 0.7749 0.8653
0.9224 026_583_00295921 Intestine 583 SW-948 456 3.908038 0.735
0.672 0.851 0.9918 1.0392 0.979 0.9178 0.9736 0.9758
026_8274_00295980 Intestine 8274 SNU-61 456 4.14154 0.663 0.72
0.672 0.6639 0.7521 0.794 0.8775 0.8761 0.9371 026_8105_00296440
Intestine 8105 LS-1034 456 4.222062 0.732 0.754 0.781 0.8085 0.8857
1.006 1.1032 1.0973 1.0698 026_580_00295930 Intestine 580 COLO-678
456 4.449849 0.891 0.793 0.767 0.8394 0.8741 0.917 1.0326 0.9344
1.0884 026_580_00266560 Intestine 580 COLO-678 456 4.550688 0.724
0.686 0.785 0.8147 0.8701 0.885 0.9546 0.9558 0.9803
026_581_00295830 Intestine 581 HCT-15 456 4.608693 0.811 0.75 0.973
0.9405 1.0972 0.98 1.098 1.0722 1.1405 026_573_00296370 Intestine
573 SW620 456 4.816766 0.775 0.787 0.797 0.8519 0.8683 0.915 0.9626
1.0901 1.097 026_8021_00296390 Intestine 8021 COLO-320-HSR 456
4.87923 0.817 0.861 0.828 1.0219 0.8974 1.103 1.0927 0.9967 1.1145
026_8106_00298521 Intestine 8106 LS-123 456 4.895207 0.706 0.749
0.852 0.7598 0.7605 0.923 0.8934 0.9319 1.0175 026_600_00296361
Intestine 600 SW 48 456 4.951306 0.706 0.776 0.775 0.7937 0.8445
0.857 0.9382 0.915 0.9609 026_8070_00296411 Intestine 8070 HCC2998
456 4.984157 0.701 0.788 0.743 0.7463 0.8151 0.891 0.9115 0.9572
0.9432 026_8135_00295950 Intestine 8135 NCI-H716 456 4.988916 0.813
0.864 0.948 0.9832 0.9658 0.94 0.9372 0.9633 1.1237
026_8136_00295961 Intestine 8136 NCI-H747 456 5.220086 0.699 0.693
0.763 0.7315 0.7863 0.815 0.8715 0.8842 1.1461 026_574_00263900
Intestine 574 CL-11 456 5.294016 0.826 0.812 0.761 0.7543 0.8087
0.891 0.9784 0.9577 0.9649 026_8026_00300671 Intestine 8026 CW-2
456 5.403672 0.827 0.955 0.974 0.9551 0.9288 0.954 0.9824 0.9866
1.0636 026_607_00293610 Intestine 607 CaR-1 456 5.551737 0.834
0.873 0.912 0.8795 0.9318 0.939 0.9636 0.9483 0.9415
026_8074_00296420 Intestine 8074 HUTU-80 456 5.701754 0.841 0.852
0.876 0.9027 0.8952 1.007 1.0026 0.966 0.9322 026_563_00316540
Intestine 563 C2BBe1 456 5.783056 0.853 0.949 0.926 0.9333 0.9253
0.879 0.9164 1.0369 1.0733 026_596_00296280 Intestine 596 MDST8 456
6.402689 0.885 0.885 0.874 0.8768 0.9374 0.931 0.9758 0.9779 1.0734
026_597_00300651 Intestine 597 SW 1116 456 6.447765 0.647 1.058
0.925 1.031 0.9839 0.997 1.0631 1.0277 0.905 026_601_00296480
Intestine 601 T84 456 7.233631 0.973 0.939 0.93 0.9736 0.9545 0.891
0.964 0.9372 1.0259 026_622_00288160 Kidney 622 G-401 456 1.158385
0.353 0.358 0.433 0.5045 0.6202 0.865 0.8778 1.0261 1.0061
026_626_00298790 Kidney 626 BFTC-909 456 1.589633 0.292 0.387 0.526
0.5522 0.8154 0.895 0.9347 1.005 1.054 026_623_00288201 Kidney 623
SK-NEP-1 456 1.837955 0.412 0.444 0.507 0.7421 0.8141 0.813 0.9099
0.9209 0.981 026_8264_00290630 Kidney 8264 RCC-JF 456 2.377362
0.452 0.535 0.585 0.6574 0.9675 0.851 0.9828 1.0591 1.0712
026_619_00290290 Kidney 619 769-P 456 2.463867 0.439 0.58 0.576
0.7655 0.8898 0.994 1.0776 1.0508 1.1513 026_627_00291130 Kidney
627 CAL-54 456 2.931844 0.586 0.588 0.649 0.6813 0.7826 0.858
0.9811 0.9726 1.0003 026_617_00290310 Kidney 617 ACHN 456 2.983353
0.545 0.592 0.627 0.7345 0.7447 0.733 0.7712 0.919 1.1152
026_8263_00290620 Kidney 8263 RCC-FG2 456 3.111338 0.517 0.689
0.691 0.7857 0.7896 0.884 0.8746 0.9754 1.0284 026_8190_00290280
Kidney 8190 TK10 456 3.318654 0.638 0.654 0.671 0.7606 0.892 0.932
1.0085 1.0009 0.9983 026_638_00288220 Kidney 638 VMRC-RCZ 456
3.36569 0.14 0.709 0.838 0.9211 0.92 0.981 0.9922 0.9588 0.989
026_8261_00308760 Kidney 8261 RCC-AB 456 3.394556 0.566 0.724 0.758
0.85 0.8258 0.948 0.9492 0.9471 0.9351 026_628_00288210 Kidney 628
SW 13 456 3.41636 0.454 0.733 0.735 0.7572 0.8068 0.826 0.8584
0.9361 1.0195 026_8262_00290610 Kidney 8262 RCC-ER 456 3.745967
0.583 0.783 0.794 0.8415 0.9082 0.87 0.8861 0.9471 0.9855
026_8265_00302360 Kidney 8265 RCC-JW 456 3.749626 0.686 0.758 0.874
0.9595 1.0107 1.003 1.0135 1.0068 1.0345 026_8261_00311200 Kidney
8261 RCC-AB 456 3.919417 0.288 0.72 0.73 0.765 0.8515 0.878 0.9287
0.8965 0.9174 026_618_00290300 Kidney 618 786-O 456 3.967789 0.657
0.719 0.822 0.8016 0.8829 0.898 1.0239 0.917 1.0472
026_625_00290670 Kidney 625 UO-31 456 3.998088 0.662 0.756 0.767
0.9002 0.8304 0.876 0.9012 1.0588 1.1507 026_8096_00290860 Kidney
8096 LB2241-RCC 456 3.99857 0.643 0.686 0.74 0.8137 0.8533 0.902
0.9235 0.9107 0.9322 026_614_00291210 Kidney 614 SW 156 456
4.137243 0.651 0.665 0.71 0.7683 0.8365 0.896 0.903 0.9074 0.9013
026_8249_00295560 Kidney 8249 NCC021 456 4.201277 0.701 0.897 0.903
0.9199 0.9735 0.942 0.9933 1.0276 1.0113 026_633_00290250 Kidney
633 KMRC-20 456 4.24311 0.713 0.872 0.83 0.9644 0.8737 1.135 1.027
1.0923 1.0373 026_8068_00290231 Kidney 8068 HA7-RCC 456 4.403879
0.761 0.764 0.7 0.8322 0.7944 0.89 1.0474 1.1224 1.0563
026_8095_00290260 Kidney 8095 LB1047-RCC 456 4.460236 0.772 0.824
0.914 1.0356 1.0154 1.004 1.0739 1.1192 1.1745 026_626_00258890
Kidney 626 BFTC-909 456 4.464205 0.741 0.932 1.057 1.0395 1.0888
1.03 1.0217 1.0625 1.0049 026_8147_00290270 Kidney 8147 OS-RC-2 456
4.480976 0.767 0.794 0.799 0.8302 0.9419 1.023 1.0718 1.0847 1.1694
026_8013_00290220 Kidney 8013 BB65-RCC 456 4.50555 0.714 0.769
0.765 0.8909 0.8509 0.865 0.9509 1.0096 0.903 026_8006_00293300
Kidney 8006 A704 456 4.630889 0.755 0.917 0.918 0.9328 0.9788 1.112
0.981 0.9813 1.0413 026_637_00290240 Kidney 637 KMRC-1 456 4.786417
0.748 0.889 0.97 0.9322 0.836 1.012 1.0239 0.9503 1.088
026_624_00290320 Kidney 624 Caki-1 456 4.808296 0.821 0.803 0.908
0.9738 0.9722 1.083 1.1676 1.1464 1.1228 026_8266_00290640 Kidney
8266 RCC-MF 456 4.811179 0.754 0.903 0.819 0.8455 0.8764 0.999
0.9885 0.935 1.1762 026_620_00288170 Kidney 620 G-402 456 4.865533
0.694 0.721 0.686 0.7919 0.7766 0.804 0.8404 0.885 0.9093
026_8152_00293381 Kidney 8152 RCC10RGB 456 5.110675 0.821 0.719
0.783 0.9032 0.8829 0.858 0.8991 0.9287 1.0343 026_640_00291220
Kidney 640 VMRC-RCW 456 5.287926 0.73 0.737 0.793 0.8088 0.8744
0.905 0.9003 0.9127 0.9163 026_8102_00293360 Kidney 8102 LB996-RCC
456 5.484542 0.855 0.901 0.954 0.9116 0.9754 0.98 0.958 0.9786
0.9921 026_1119_00290901 Kidney 1119 SN-12C 456 5.586587 0.818
0.749 0.747 0.783 0.8058 0.856 0.9038 0.9508 0.9573
026_626_00290810 Kidney 626 BFTC-909 456 5.753321 0.799 0.854 0.903
0.9293 0.9289 0.92 0.9333 0.9322 0.9535 026_8005_00266530 Kidney
8005 A498 456 5.843211 0.594 0.917 1.009 1.0079 0.9307 0.996 1.0002
1.0033 0.9941 026_8157_00296471 Kidney 8157 RXF393 456 6.166218
0.853 0.878 0.91 0.8924 0.9071 0.932 0.9081 0.9505 0.9889
026_8102_00253000 Kidney 8102 LB996-RCC 456 6.306723 0.959 0.873
0.878 0.9741 0.9596 1.025 0.9525 0.9849 0.9931 026_8006_00263880
Kidney 8006 A704 456 6.969306 1.351 0.929 1.157 0.9881 1.0206 0.957
1.0009 0.9583 0.9931 026_8152_00256190 Kidney 8152 RCC10RGB 456
7.15535 1.178 0.835 1.176 0.846 0.8683 1.123 1.1733 1.1056 1.0876
026_8005_00296380 Kidney 8005 A498 456 8.190783 1.022 1.025 1.024
1.0158 1.03 1.032 1.0128 1.0239 1.0104 026_233_00277420 Leukemia
233 SIG-M5 456 -5.883853 0.228 0.243 0.233 0.2401 0.2362 0.247
0.2641 0.2732 0.4405 026_217_00277380 Leukemia 217 OCI-AML2 456
-1.42786 0.09 0.102 0.106 0.1256 0.1044 0.132 0.3277 0.8333 0.8618
026_179_00314500 Leukemia 179 KMOE-2 456 -0.249088 0.13 0.135 0.138
0.1591 0.3658 0.608 0.865 0.8967 1.0615 026_214_00285590 Leukemia
214 NB-4 456 -0.011745 0.18 0.323 0.315 0.3611 0.4215 0.592 0.8144
0.8865 0.9464 026_168_00280410 Leukemia 168 JURL-MK1 456 0.181843
0.145 0.179 0.186 0.2526 0.4638 0.727 0.9379 0.904 0.9907
026_194_00280680 Leukemia 194 ML-2 456 0.294246 0.418 0.395 0.379
0.4439 0.4739 0.633 0.7874 0.8519 1.0223 026_186_00280670 Leukemia
186 LAMA-84 456 0.295979 0.152 0.15 0.191 0.3623 0.6147 0.774 0.777
0.8716 0.8816 026_234_00314650 Leukemia 234 SKM-1 456 0.349518 0.37
0.347 0.369 0.4439 0.4995 0.57 0.729 0.9566 1.2103 026_221_00280300
Leukemia 221 OCI-M1 456 0.656393 0.183 0.235 0.344 0.5052 0.5942
0.759 0.8267 0.8418 1.1212 026_260_00280430 Leukemia 260 KO52 456
0.674413 0.348 0.376 0.357 0.3446 0.4195 0.753 0.7217 0.7637 1.0996
026_45_00274530 Leukemia 45 HL-60 456 0.756274 0.344 0.389 0.403
0.4669 0.6154 0.78 0.9211 0.9579 1.0107 026_218_00279140 Leukemia
218 OCI-AML3 456 0.868641 0.368 0.38 0.401 0.4571 0.5459 0.76
0.8359 0.9311 0.9403 026_219_00280290 Leukemia 219 OCI-AML5 456
0.879404 0.245 0.404 0.427 0.4943 0.686 0.712 0.8112 0.9498 0.9021
026_199_00314510 Leukemia 199 MOLT-13 456 0.892109 0.227 0.339
0.454 0.5097 0.5774 0.766 0.8101 0.9633 1.1416 026_226_00280310
Leukemia 226 PL-21 456 0.938066 0.406 0.425 0.463 0.4834 0.537
0.657 0.7338 0.7978 0.8482 026_8141_00274380 Leukemia 8141 NOMO-1
456 1.040856 0.375 0.445 0.435 0.4798 0.5465 0.636 0.9597 0.9214
0.9288 026_8069_00279180 Leukemia 8069 HAL-01 456 1.147185 0.349
0.345 0.454 0.634 0.6764 1.103 0.7322 0.8261 1.0097 026_68_00273640
Leukemia 68 MV-4-11 456 1.155322 0.061 0.367 0.445 0.5488 0.6586
0.777 0.8697 0.9167 1.045 026_175_00282940 Leukemia 175 KARPAS-620
456 1.190853 0.562 0.517 0.444 0.6051 0.7411 0.916 0.9551 1.1003
1.0193 026_89_00278880 Leukemia 89 MEG-01 456 1.211836 0.223 0.376
0.433 0.6062 0.6931 0.86 0.9075 0.9993 1.0376 026_8017_00279150
Leukemia 8017 BV-173 456 1.464827 0.147 0.454 0.57 0.8503 0.5534
0.638 0.7679 0.8629 1.0089 026_225_00277400 Leukemia 225 PF-382 456
1.646942 0.317 0.403 0.525 0.6985 0.7396 0.932 0.9611 0.9018 1.0833
026_285_00282960 Leukemia 285 KY821A3 456 1.656661 0.422 0.426
0.445 0.6672 0.7837 0.935 1.0015 1.0293 1.0583 026_8008_00280620
Leukemia 8008 ALL-PO 456 1.659142 0.143 0.469 0.549 0.6225 0.6911
0.725 0.845 0.8482 1.019
026_177_00276860 Leukemia 177 KE-37 456 1.711407 0.195 0.451 0.523
0.6745 0.9068 0.897 0.9087 0.9169 1.0477 026_126_00280270 Leukemia
126 GDM-1 456 1.723253 0.297 0.444 0.595 0.682 0.7944 0.786 0.9053
0.9081 0.8666 026_261_00274440 Leukemia 261 MY-M12 456 1.811821
0.395 0.471 0.59 0.6443 0.7461 0.832 0.8778 1.0023 1.0569
026_201_00277390 Leukemia 201 MOLT-16 456 1.821177 0.276 0.43 0.578
0.7394 0.7801 0.857 0.9379 1.0322 0.9678 026_148_00277330 Leukemia
148 CMK 456 1.894859 0.275 0.484 0.637 0.6101 0.744 0.771 0.9002
0.8949 0.8668 026_28_00280320 Leukemia 28 SUP-B15 456 1.921651
0.261 0.506 0.579 0.7214 0.7165 0.82 0.908 0.9679 0.9913
026_190_00287920 Leukemia 190 ME-1 456 1.924633 0.611 0.596 0.587
0.5806 0.5987 0.684 0.7906 0.8581 0.9106 026_8150_00273700 Leukemia
8150 QIMR-WIL 456 1.928333 0.441 0.546 0.595 0.6574 0.7011 0.771
0.852 0.962 0.9148 026_8156_00273710 Leukemia 8156 RPMI-8866 456
1.939976 0.339 0.474 0.598 0.694 0.7428 0.867 0.9518 0.9936 0.9682
026_161_00277350 Leukemia 161 HC-1 456 1.948766 0.447 0.517 0.568
0.6165 0.6342 0.768 0.7642 0.9074 1.0155 026_195_00279120 Leukemia
195 MOLM-13 456 1.961596 0.394 0.455 0.518 0.6868 0.817 0.872
0.9157 0.983 1.0345 026_209_00279130 Leukemia 209 NALM-6 456
2.000316 0.14 0.471 0.592 0.6915 0.7587 0.808 0.8658 0.9205 0.9768
026_127_00278810 Leukemia 127 CESS 456 2.000616 0.342 0.505 0.625
0.6869 0.7679 0.843 0.9355 0.9565 0.9827 026_8196_00278790 Leukemia
8196 697 456 2.032634 0.337 0.476 0.617 0.7249 0.8167 0.885 1.0481
1.0695 1.0723 026_223_00276841 Leukemia 223 P12-ICHIKAWA 456
2.094412 0.266 0.472 0.62 1.0139 0.9551 0.864 0.9219 1.0283 0.907
026_157_00279291 Leukemia 157 DND-41 456 2.109279 0.354 0.498 0.624
0.7174 0.8189 0.89 0.9495 0.9928 1.0144 026_174_00276850 Leukemia
174 KARPAS-45 456 2.156973 0.368 0.541 0.63 0.778 0.8566 0.998
1.0142 0.9561 0.9985 026_223_00274461 Leukemia 223 P12-ICHIKAWA 456
2.170641 0.274 0.465 0.567 0.909 0.9914 1.041 1.0272 0.9908 1.0038
026_231_00277410 Leukemia 231 RPMI-8402 456 2.171412 0.292 0.525
0.673 0.8379 0.894 0.956 1.0525 1.0069 1.0163 026_176_00278840
Leukemia 176 KCL-22 456 2.178418 0.342 0.551 0.66 0.7185 0.7521
0.822 0.8942 1.034 0.9817 026_198_00279320 Leukemia 198 MOLP-8 456
2.182175 0.422 0.613 0.604 0.6789 0.7123 0.753 0.8469 0.8678 0.9555
026_35_00274541 Leukemia 35 MOLT-4 456 2.185513 0.368 0.518 0.657
0.7557 0.8266 0.893 0.9057 1.0649 1.0505 26_256_00273800 Leukemia
256 U266B1 456 2.191112 0.423 0.53 0.596 0.6351 0.7316 0.782 0.8701
0.9616 0.9089 026_41_00278850 Leukemia 41 KG-1 456 2.260524 0.445
0.507 0.569 0.7321 0.8443 0.89 0.9006 0.9237 0.9648 026_38_00278900
Leukemia 38 THP-1 456 2.296961 0.389 0.568 0.692 0.7667 0.7899
0.895 0.9332 0.9649 0.9902 026_153_00277340 Leukemia 153 CTV-1 456
2.301756 0.308 0.563 0.682 0.7882 0.8949 0.972 1.0362 1.0213 1.0568
26_284_00273770 Leukemia 284 KY821 456 2.31293 0.459 0.524 0.599
0.642 0.7677 0.831 0.9119 0.9596 0.9938 026_256_00304780 Leukemia
256 U266B1 456 2.374209 0.453 0.543 0.608 0.6629 0.7604 0.806
0.9117 0.9547 0.9487 026_8033_00279160 Leukemia 8033 DEL 456
2.390133 0.433 0.595 0.647 0.7617 0.7648 0.851 0.9096 0.9116 0.9641
026_227_00314640 Leukemia 227 RCH-ACV 456 2.405281 0.39 0.571 0.742
0.8517 0.9043 0.946 0.9917 1.0175 1.1197 026_33_00274521 Leukemia
33 CCRF-CEM 456 2.408403 0.354 0.606 0.685 0.7594 0.8191 0.904
0.9597 0.9629 1.0269 026_141_00276820 Leukemia 141 K-562 456
2.457405 0.51 0.504 0.624 0.8081 0.8379 1.163 0.9404 1.0057 1.0193
026_8137_00273680 Leukemia 8137 NKM-1 456 2.543263 0.464 0.601
0.622 0.6486 0.7425 0.801 0.8993 0.9038 0.9449 026_36_00274550
Leukemia 36 Reh 456 2.569808 0.418 0.628 0.742 0.8367 0.88 0.896
0.9262 0.8979 0.9849 026_8042_00285400 Leukemia 8042 EoL-1 -cell
456 2.64235 0.232 0.569 0.651 0.7409 0.7743 0.894 0.8333 0.905
0.9094 026_281_00273690 Leukemia 281 P30/OHK 456 2.804566 0.473
0.654 0.769 0.9121 0.8865 1.022 1.0746 1.0233 1.0407
026_59_00278820 Leukemia 59 J.RT3-T3.5 456 2.830428 0.457 0.659
0.795 0.8429 0.9259 1.002 1.0587 0.9873 1.1396 026_164_00277360
Leukemia 164 HEL 456 2.835925 0.426 0.745 0.939 0.9081 1.0194 0.987
1.0611 1.0159 0.9841 026_183_00277370 Leukemia 183 L-363 456
2.864138 0.468 0.685 0.82 0.9025 0.9548 1.046 1.0249 0.9812 1.02
026_90_00282840 Leukemia 90 KU812 456 2.874545 0.446 0.74 0.806
0.917 0.9589 0.978 0.9646 1.0111 1.0409 026_27_00278890 Leukemia 27
RS4; 11 456 2.914096 0.47 0.684 0.809 0.8895 0.9144 0.975 0.9547
0.9619 1.0194 026_167_00314490 Leukemia 167 JURKAT 456 2.954834
0.503 0.713 0.842 0.9399 0.9544 0.985 0.9987 1.034 1.0702
026_181_00278860 Leukemia 181 KOPN-8 456 2.957727 0.484 0.669 0.731
0.7843 0.8662 0.92 0.9473 0.9339 0.9555 026_8155_00309110 Leukemia
277 RPMI 8226 456 2.959542 0.648 0.63 0.689 0.8376 0.888 0.99
0.9938 1.0182 1.0157 026_8041_00280640 Leukemia 8041 EM-2 456
3.022652 0.557 0.687 0.614 0.634 0.706 0.679 0.8613 0.8617 1.9538
026_142_00279280 Leukemia 142 ALL-SIL 456 3.085854 0.187 0.624
0.769 0.7092 0.7975 0.937 0.9525 0.9717 0.8276 026_277_00280420
Leukemia 277 RPMI 8226 456 3.102844 0.659 0.636 0.566 0.6117 0.6594
0.722 0.8693 0.9508 0.9057 026_180_00279310 Leukemia 180 KMS-12-BM
456 3.125028 0.512 0.704 0.788 0.8503 0.8703 0.949 0.905 0.9806
0.9931 026_8066_00279170 Leukemia 8066 GR-ST 456 3.192114 0.532
0.681 0.762 0.8318 0.8448 0.865 1.0258 0.9609 0.9989
26_283_00273780 Leukemia 283 LC4-1 456 3.243599 0.592 0.732 0.916
0.9522 1.0268 1.078 0.9517 1.0295 1.0135 026_114_00273650 Leukemia
114 SUP-T1 456 3.397019 0.637 0.807 0.908 1.0154 0.984 1.031 0.99
0.9877 1.0134 026_138_00287901 Leukemia 138 Loucy 456 3.501279
0.289 0.749 0.772 0.942 0.9225 0.948 0.9678 0.9653 1.0654
026_8014_00282911 Leukemia 8014 BE-13 456 3.615529 0.68 0.757 0.965
1.1246 1.0528 1.075 1.0884 1.0694 1.1037 26_274_00273860 Leukemia
274 BALL-1 456 3.704228 0.65 0.709 0.777 0.8768 0.9725 0.983 0.9819
0.9839 0.9978 026_222_00280480 Leukemia 222 OPM-2 456 3.757754
0.681 0.619 0.771 0.9443 0.8789 1.073 0.9014 0.9191 0.9051
026_8164_00283540 Leukemia 8164 SK-MM-2 456 3.790344 0.669 0.788
0.861 0.8903 0.938 0.941 1.0087 0.9277 0.9972 026_166_00278830
Leukemia 166 JJN-3 456 3.901981 0.636 0.791 0.85 0.8566 0.8711
0.916 0.9455 0.9892 1.057 026_230_00282970 Leukemia 230 ROS-50 456
4.130585 0.741 0.775 0.805 0.9413 0.9813 1.11 1.1334 1.0559 0.999
026_159_00314480 Leukemia 159 EJM 456 4.251735 0.714 0.845 0.899
0.9099 1.0285 0.923 1.0677 1.0768 1.0543 026_278_00304730 Leukemia
278 KMS-12-PE 456 4.269602 0.765 0.814 0.84 1.0641 0.988 1.023
1.0045 1.0284 1.0665 026_8219_00282850 Leukemia 8219 Mo-T 456
4.313664 0.73 0.825 0.804 0.9425 0.9233 1.033 0.8454 1.1551 1.0561
026_279_00274450 Leukemia 279 P31/FUJ 456 4.386897 0.705 0.805
0.799 0.819 0.8632 1.043 1.0364 0.9822 1.0363 026_171_00285190
Leukemia 171 KARPAS-231 456 4.404825 0.703 0.9 0.906 0.9339 0.9295
0.904 0.9725 0.9496 0.9164 026_244_00273660 Leukemia 244 TALL-1 456
4.411016 0.698 0.758 0.785 0.8345 0.8593 0.861 1.0445 1.0026 1.0194
026_158_00291340 Leukemia 158 EHEB 456 4.509743 0.665 0.745 0.765
0.7841 0.8282 0.839 0.92 0.9348 1.0344 026_134_00278800 Leukemia
134 ARH-77 456 4.545002 0.781 0.856 0.96 0.848 1.0568 1.029 1.0104
1.0156 1.0086 026_246_00273670 Leukemia 246 U-698-M 456 4.580953
0.754 0.891 0.936 0.9419 0.9607 0.937 1.032 0.9352 1.0229
026_8113_00280440 Leukemia 8113 MHH-CALL-2 456 4.648418 0.771 0.883
0.929 0.9704 0.9337 0.987 1.0345 1.0223 1.0471 026_159_00311690
Leukemia 159 EJM 456 4.699945 0.746 0.894 0.921 0.9275 0.9393 0.941
0.9575 0.9338 0.9675 026_159_00282920 Leukemia 159 EJM 456 4.832393
0.746 0.976 1.016 0.9859 1.0015 1.002 1.1587 1.1041 1.0548
026_8115_00280450 Leukemia 8115 MN-60 456 4.861437 0.826 0.804
0.822 1.0586 0.9555 0.947 1.0471 1.128 1.0314 026_204_00280460
Leukemia 204 MONO-MAC-6 456 4.946039 0.736 0.802 0.771 0.8409
0.8712 0.833 0.8634 1.0677 0.855 026_8117_00280470 Leukemia 8117
MUTZ-1 456 5.304526 0.823 0.984 1.063 1.051 1.0142 1.007 0.9986
0.9654 0.9525 026_188_00282930 Leukemia 188 LP-1 456 5.373997 0.831
0.887 0.972 0.8808 0.9105 0.886 0.9533 0.9576 1.138
026_8081_00285580 Leukemia 8081 JVM-3 456 5.581328 0.884 0.904
0.907 0.9131 1.01 1.097 1.0593 0.9629 1.0333 026_159_00309070
Leukemia 159 EJM 456 5.810283 0.882 0.986 0.998 1.0992 1.0048 1.045
1.0126 1.024 1.0649 026_8080_00280660 Leukemia 8080 JVM-2 456
5.982881 0.867 0.876 0.901 0.8865 0.9235 0.909 0.9782 0.8964 0.9789
026_278_00306920 Leukemia 278 KMS-12-PE 456 6.629507 0.971 0.92
0.95 0.9554 0.9046 0.905 0.9657 1.0243 0.9236 026_8010_00280630
Leukemia 8010 ATN-1 456 7.14863 1.03 0.958 0.835 0.8896 0.8651
0.907 0.9392 0.8582 0.9763 26_262_00273790 Leukemia 262 MLMA 456
7.68552 0.485 1.033 1.077 1.0216 1.0341 0.962 1.0378 0.9821 1.0245
026_278_00282950 Leukemia 278 KMS-12-PE 456 7.74945 0.958 1.101
1.04 1.1177 1.0046 1.029 1.0272 1.0311 1.0037 026_649_00264910
Liver 649 Hep 3B2.1-7 456 -0.30042 0.134 0.098 0.087 0.1121 0.2203
0.601 0.8432 0.8926 0.9089 026_658_00262810 Liver 658 JHH-1 456
-0.185127 0.054 0.329 0.515 0.3993 0.5194 0.524 0.6608 0.6499
0.6863 026_649_00266180 Liver 649 Hep 3B2.1-7 456 -0.037335 0.073
0.105 0.093 0.0908 0.1037 0.202 0.8123 0.9207 1.007
026_667_00273550 Liver 667 HuH-7 456 1.289162 0.279 0.565 0.539
0.5535 0.7134 0.77 0.8626 0.9675 0.9662 026_659_00255780 Liver 659
JHH-2 456 1.758427 0.283 0.473 0.657 0.7149 0.551 0.819 0.8858
0.8891 1.0202 026_643_00266460 Liver 643 SNU-398 456 2.578054 0.415
0.687 0.643 0.6854 0.6879 0.717 0.7896 0.8076 0.9193
026_667_00269210 Liver 667 HuH-7 456 2.760591 0.501 0.59 0.665
0.6852 0.6886 0.781 0.95 1.0287 1.0142 026_647_00269110 Liver 647
SNU-387 456 2.930627 0.494 0.772 0.829 0.9402 0.9686 1.046 1.0372
1.0223 1.2144 026_661_00252500 Liver 661 JHH-7 456 3.286573 0.539
0.889 0.794 0.8777 1.0776 0.953 0.9918 1.037 0.9476
026_643_00263980 Liver 643 SNU-398 456 3.293137 0.487 0.768 0.854
0.8235 0.8763 0.883 0.9616 1.068 1.0718 026_642_00308440 Liver 642
C3A 456 3.554832 0.587 0.68 0.706 0.7687 0.817 0.878 0.9697 0.9574
0.9245 026_648_00258350 Liver 648 SNU-423 456 3.576003 0.139 0.699
0.902 0.9157 0.9485 0.909 0.9218 0.9469 1.0028 026_656_00252510
Liver 656 JHH-4 456 3.61979 0.701 0.653 0.631 0.8187 0.7968 0.977
0.9161 0.9897 0.9288 026_660_00252490 Liver 660 JHH-6 456 3.831185
0.696 0.88 1.034 0.9294 0.9465 1.04 0.9417 1.0528 0.952
026_644_00252750 Liver 644 SNU-449 456 4.40481 0.664 0.733 0.764
0.7954 0.8597 0.896 0.9485 0.9457 0.9162 026_644_00306170 Liver 644
SNU-449 456 6.450644 0.96 0.961 0.745 1.0716 1.0197 1.08 0.991
0.9007 0.9369 26_646_00314060 Liver 646 SNU-475 456 6.501654 0.181
0.97 0.971 0.9863 1.0061 1.007 1.0271 1.0202 0.057 026_654_00255800
Liver 654 SK-HEP-1 456 6.515068 0.927 0.956 0.961 0.9827 0.9589
0.98 0.9783 0.9726 0.9998 026_668_00252690 Liver 668 HLE 456
7.25378 0.434 1.006 1.087 0.9897 1.1103 0.964 1.006 1.106 0.9541
026_662_00252460 Liver 662 huH-1 456 7.722693 0.466 1.165 1.072
1.0401 1.0408 1.136 1.015 1.123 0.9778 026_645_00306160 Liver 645
SNU-182 456 7.852049 1.105 1.061 1.063 1.0055 1.0268 1.053 1.0365
1.0318 1.0842 026_642_00252670 Liver 642 C3A 456 7.864415 0.386 1.1
1.007 1.108 0.9463 1.051 0.9803 1.029 0.9218 026_830_00304760 Lung
830 NCI-H2135 456 -0.374655 0.298 0.262 0.246 0.3049 0.3692 0.528
0.7269 0.8905 0.977 026_698_00300170 Lung 698 NCI-H524 456 2.279246
0.443 0.506 0.63 0.8741 0.9302 1.005 1.0131 1.0304 1.0567
026_672_00314460 Lung 672 NCI-H510A 456 2.367223 0.323 0.585 0.763
0.783 0.8906 0.969 0.9972 1.1809 1.0131 026_761_00300410 Lung 761
COR-L279 456 3.131703 0.559 0.745 0.868 0.9821 1.0079 1.06 1.0144
1.0182 1.0709 026_726_00304770 Lung 726 NCI-H2171 456 3.185965
0.524 0.729 0.842 0.8655 0.9088 0.871 1.0252 0.9207 1.129
026_740_00302760 Lung 740 NCI-H82 456 3.210581 0.542 0.755 0.865
0.9592 1.0522 0.888 0.9137 1.004 1.0151 026_787_00302910 Lung 787
SBC-3 456 3.410219 0.582 0.655 0.694 0.802 0.8502 0.902 0.9485
0.9451 0.9885 026_695_00300150 Lung 695 NCI-H211 456 3.515811 0.644
0.645 0.719 0.8327 0.8177 0.933 0.9519 0.9812 1.0305
026_721_00302860 Lung 721 NCI-H2029 456 3.591515 0.606 0.8 0.891
0.9011 0.8579 0.931 0.9429 1.0032 1.0011 026_776_00303250 Lung 776
MS-1-L 456 3.614256 0.642 0.767 0.855 0.8458 1.1115 0.929 1.0414
1.0285 1.162 026_8197_00304741 Lung 8197 LU-139 456 3.667637 0.66
0.84 0.901 1.0158 0.9097 0.912 1.0844 1.1048 1.0641
026_702_00302900 Lung 702 NCI-H847 456 3.686524 0.818 0.707 0.668
0.6815 0.7184 0.743 0.7999 0.9145 0.9524 026_8203_00309050 Lung
8203 IST-SL1 456 3.697133 0.635 0.801 0.87 0.931 0.9112 1 1.0127
0.9507 1.0704 026_724_00300140 Lung 724 NCI-H2081 456 3.857634
0.623 0.866 0.895 0.9027 0.9349 0.948 0.969 0.9681 1.0072
026_765_00300050 Lung 765 DMS 273 456 3.890251 0.702 0.844 0.904
0.9648 1.028 0.968 1.0795 1.0165 1.0825 026_829_00305160 Lung 829
NCI-H2110 456 3.972695 0.67 0.694 0.776 0.8765 0.8765 0.946 1.0573
0.91 0.9398 026_742_00303230 Lung 742 DMS 53 456 3.990104 0.711
0.802 0.867 0.9734 1.0075 1.037 1.0307 1.0204 1.0405
026_710_00316710 Lung 710 NCI-H1341 456 4.024081 0.591 0.852 0.7
0.7407 0.8259 0.789 0.8069 0.8386 0.865 026_738_00302800 Lung 738
NCI-H446 456 4.043153 0.692 0.871 0.917 1.0421 0.9341 0.915 1.0657
0.9649 1.0382 026_751_00308570 Lung 751 NCI-H209 456 4.061511 0.657
0.848 0.89 0.9403 0.8939 0.91 0.9316 0.9524 0.9513 026_716_00298900
Lung 716 NCI-H1876 456 4.083964 0.744 0.866 0.936 0.9448 0.9881
0.979 0.9782 0.9994 1.0261 026_725_00303280 Lung 725 NCI-H2141 456
4.09696 0.711 0.842 0.928 0.9702 0.9155 0.924 1.0044 1.0101 0.9559
026_688_00302810 Lung 688 SW 1271 456 4.124557 0.602 0.761 0.79
0.8093
0.807 0.823 0.896 0.8782 0.9341 026_720_00302380 Lung 720 NCI-H1994
456 4.130367 0.696 0.694 0.73 0.8114 0.9245 0.935 0.9484 0.9393
0.9738 026_811_00311720 Lung 811 NCI-H1435 456 4.138326 0.635 0.803
0.855 0.8503 0.8571 0.89 0.8912 0.955 0.9956 026_704_00300250 Lung
704 NCI-H1048 456 4.178908 0.166 0.762 0.867 0.8914 0.9318 0.923
0.9201 1.0122 0.9635 026_746_00302780 Lung 746 SHP-77 456 4.20487
0.79 0.843 0.955 1.029 0.9896 1.021 1.0111 0.9931 1.0025
026_829_00311740 Lung 829 NCI-H2110 456 4.22315 0.739 0.83 0.914
0.9675 0.9597 0.983 0.9791 1.032 0.9995 026_736_00300181 Lung 736
NCI-H69 456 4.26966 0.798 0.87 0.945 1.0535 1.0472 1.143 1.0521
1.0846 1.1743 026_724_00303270 Lung 724 NCI-H2081 456 4.274134
0.708 0.95 1.023 0.9666 1.0134 0.99 1.0037 1.0388 1.0531
026_714_00300260 Lung 714 NCI-H1694 456 4.276996 0.701 0.831 0.843
0.8708 0.9426 0.93 0.9095 1.1071 1.0848 026_715_00298890 Lung 715
NCI-H1836 456 4.291554 0.73 0.833 0.997 0.9315 0.9434 0.955 0.9291
0.9423 1 026_757_00302870 Lung 757 CPC-N 456 4.345126 0.718 0.819
0.887 0.8883 0.9167 0.938 0.9641 1.0154 1.0153 026_8229_00304990
Lung 8229 COR-L303 456 4.468573 0.685 0.828 0.788 0.868 0.6881
0.855 0.9579 0.8466 0.9772 026_691_00308560 Lung 691 NCI-H526 456
4.602758 0.748 0.932 0.944 0.9861 0.9744 0.954 1.0035 1.0955 1.0007
026_8099_00316740 Lung 8099 LB647-SCLC 456 4.613258 0.755 0.954
0.998 0.9372 0.957 1.041 1.0171 1.0259 0.9506 026_725_00302750 Lung
725 NCI-H2141 456 4.618451 0.747 0.869 0.906 0.9009 0.9283 0.901
1.003 0.9238 1.1424 026_705_00314520 Lung 705 NCI-H1092 456
4.678169 0.77 0.861 0.911 1.02 0.9158 1.082 0.9997 0.9139 0.961
026_684_00303260 Lung 684 NCI-H1688 456 4.865947 0.832 0.831 0.988
1.0672 0.9906 1.099 1.0391 1.0378 1.0698 026_725_00300160 Lung 725
NCI-H2141 456 4.894458 0.785 0.908 0.936 0.9468 0.9309 0.94 0.9635
1.0477 0.9963 026_8281_00300930 Lung 8281 COR-L311 456 5.024667
0.792 0.892 0.947 0.9155 0.8604 0.871 1.0268 1.0989 1.045
026_757_00300940 Lung 757 CPC-N 456 5.046784 0.796 0.904 0.984
1.0041 1.0137 0.919 0.905 0.9063 1.0167 026_701_00309060 Lung 701
NCI-H841 456 5.157768 0.826 0.808 0.939 0.891 0.8811 0.919 0.9514
1.0474 1.0933 026_814_00304750 Lung 814 NCI-H1568 456 5.159278
0.784 0.726 0.739 0.745 0.7127 0.774 0.9411 0.9 0.9817
026_741_00305010 Lung 741 NCI-H345 456 5.184481 0.801 0.818 0.835
0.9503 0.9116 0.894 0.8849 0.9853 1.083 026_705_00311700 Lung 705
NCI-H1092 456 5.191642 0.849 0.892 0.945 1.0426 0.9184 0.981 1.0366
1.0158 1.0581 026_786_00300950 Lung 786 SBC-5 456 5.281883 0.841
1.002 0.991 1.0148 0.9974 0.998 1.0298 1.0075 1.0015
026_723_00316720 Lung 723 NCI-H2066 456 5.313778 0.793 0.815 0.866
0.86 0.9117 0.892 0.9084 0.923 0.9757 026_705_00309080 Lung 705
NCI-H1092 456 5.329516 0.829 0.941 0.999 0.9672 0.8963 0.935 0.9991
0.9888 1.0506 026_709_00305000 Lung 709 NCI-H1304 456 5.330039
0.838 0.882 0.906 0.8939 0.9384 0.978 1.0127 1.0322 1.0575
026_739_00314630 Lung 739 NCI-H146 456 5.38813 0.858 0.893 0.978
1.0115 1.0583 0.937 0.9685 1.0018 1.0831 026_8110_00314450 Lung
8110 LU-165 456 5.539798 0.87 0.909 0.986 0.9964 0.9809 1.094
0.9305 0.9513 1.1416 026_811_00305140 Lung 811 NCI-H1435 456
5.586661 0.886 0.953 0.978 0.9914 1.0102 0.994 0.9871 0.9783 0.9731
026_711_00311710 Lung 711 NCI-H1417 456 5.606717 0.875 0.91 0.911
0.9612 1.0374 0.917 1.267 1.0195 0.9447 026_712_00309090 Lung 712
NCI-H1436 456 5.636542 0.85 0.918 0.976 0.9527 0.9331 0.932 0.9583
0.9477 1.0343 026_711_00305130 Lung 711 NCI-H1417 456 5.804325
0.849 1.022 0.916 0.9247 0.9771 0.939 0.9326 1.0168 1.0143
026_728_00311760 Lung 728 NCI-H2196 456 5.845371 0.89 0.908 0.918
0.9801 0.875 1.033 0.9519 0.931 1.0431 026_831_00311750 Lung 831
NCI-H2172 456 5.851811 0.879 0.986 1.002 0.9709 0.9562 0.984 0.9781
1.0144 0.974 026_1216_00300060 Lung 1216 H292 456 5.909493 0.869
0.823 0.808 0.8192 0.8429 0.864 0.9245 0.9789 1.0777
026_8109_00314440 Lung 8109 LU-134-A 456 5.930549 0.868 0.95 0.972
0.9375 0.9332 0.959 0.9516 1.1207 1.1025 026_831_00305170 Lung 831
NCI-H2172 456 6.053261 0.837 0.856 0.908 0.8908 0.9397 0.905 0.9677
0.9512 0.9839 026_728_00305180 Lung 728 NCI-H2196 456 6.171009 0.9
0.903 0.976 0.9979 0.9199 0.928 0.9453 0.9369 0.941
026_689_00300910 Lung 689 NCI-H187 456 6.191556 0.887 1.009 0.976
0.9779 0.9535 1.047 1.021 1.0846 0.959 026_785_00302770 Lung 785
SBC-1 456 6.234067 0.875 1.037 1.061 0.9376 1.0385 0.937 0.9403
1.0039 1.1602 026_8280_00306910 Lung 8280 COR-L321 456 6.254083
0.965 0.937 1.019 0.8492 0.948 1.048 1.0366 1.0761 0.9586
026_706_00318720 Lung 706 NCI-H1105 456 6.355 0.897 0.965 1.009
0.9821 0.9646 1.037 0.9974 0.9667 0.9589 026_712_00305150 Lung 712
NCI-H1436 456 6.498513 0.959 0.818 0.938 1.1033 0.9924 0.922 1.0128
1.0035 0.9263 026_8079_00306720 Lung 8079 IST-SL2 456 6.56418 0.947
0.972 0.935 0.9658 0.9445 0.966 0.9554 1.0191 1.0006
026_743_00303220 Lung 743 DMS 114 456 6.614771 0.862 0.853 0.785
0.7831 0.861 0.851 0.8815 0.9129 1.0374 026_8022_00306711 Lung 8022
COLO-668 456 6.912523 0.97 0.968 0.956 1 0.9608 0.917 1.0093 0.9217
0.9308 026_8109_00308510 Lung 8109 LU-134-A 456 6.98132 0.924 1.012
1.105 0.9946 0.9684 1.081 0.9878 0.9397 0.9606 026_758_00303240
Lung 758 HCC-33 456 7.022636 0.931 1.056 1.002 1.0822 1.0508 1.011
1.0081 1.1612 1.1932 026_771_00306940 Lung 771 Lu-135 456 7.040802
0.988 0.969 0.972 1.0271 0.9989 1.006 0.993 1.0649 1.0235
026_694_00302790 Lung 694 NCI-H196 456 7.041262 0.976 0.954 0.975
0.9548 0.9357 0.922 0.9564 0.9433 1.019 026_764_00308550 Lung 764
COR-L95 456 7.251052 0.978 1.021 0.944 1.002 1.0708 0.947 0.9637
0.9383 0.903 026_763_00300900 Lung 763 COR-L88 456 7.365052 0.96
1.168 0.891 1.1434 1.1503 0.976 1.0066 1.0636 1.052
026_8134_00306730 Lung 8134 NCI-H64 456 7.782872 1.026 1.003 0.976
1.0025 0.951 1.047 0.9338 0.8997 1.0076 026_712_00311730 Lung 712
NCI-H1436 456 7.804947 1.019 1.049 0.947 0.9403 0.9104 0.931 0.9349
0.9472 0.9659 026_8018_00304801 Lung: NSCLC 8018 Calu-6 456
-0.899538 0.106 0.165 0.183 0.2075 0.2699 0.394 0.8182 0.9121
1.0531 026_1246_00304570 Lung: NSCLC 1246 NCI-H1770 456 -0.072444
0.109 0.175 0.267 0.3543 0.4184 0.552 0.6475 1.0183 0.9838
026_847_00304580 Lung: NSCLC 847 NCI-H2087 456 1.048994 0.368 0.402
0.458 0.4756 0.5065 0.683 0.7888 0.9774 0.9846 026_680_00298830
Lung: NSCLC 680 NCI-H727 456 1.334143 0.329 0.434 0.475 0.4492
0.5064 0.604 0.8522 0.9134 0.98 026_748_00304590 Lung: NSCLC 748
NCI-H226 456 1.690128 0.342 0.457 0.514 0.5901 0.7687 0.757 0.7854
1.0039 0.932 026_851_00298380 Lung: NSCLC 851 CAL-12T 456 1.708943
0.452 0.449 0.49 0.5839 0.7337 0.842 0.9205 0.938 0.9948
026_861_00300230 Lung: NSCLC 861 LCLC-97TM1 456 2.249036 0.424
0.573 0.622 0.7209 0.7517 0.873 1.0412 0.9724 0.974
026_1245_00304550 Lung: NSCLC 1245 NCI-H1648 456 2.511131 0.507
0.571 0.662 0.5623 0.7611 0.961 1.0696 1.0483 1.4204
026_1180_00308140 Lung: NSCLC 1180 NCI-H3122 456 2.550953 0.534
0.517 0.616 0.6044 0.7162 0.883 0.9512 0.9659 0.9657
026_802_00298451 Lung: NSCLC 802 NCI-H358 456 2.662533 0.509 0.53
0.626 0.7229 0.8072 0.806 0.8743 0.9218 0.9534 026_815_00311140
Lung: NSCLC 815 NCI-H1623 456 2.802368 0.523 0.592 0.685 0.6168
0.6264 0.746 0.872 0.8493 1.0805 026_1180_00302350 Lung: NSCLC 1180
NCI-H3122 456 2.945291 0.724 0.649 0.605 0.75 0.7455 0.913 0.9083
1.0538 1.1763 026_8040_00304501 Lung: NSCLC 8040 EKVX 456 3.134434
0.532 0.642 0.782 0.752 0.7074 0.804 0.9087 1.0521 1.0032
026_865_00308451 Lung: NSCLC 865 COR-L23 456 3.225036 0.58 0.657
0.735 0.83 0.9489 0.935 1.0144 1.0355 1.0002 026_1243_00304541
Lung: NSCLC 1243 NCI-H1395 456 3.23207 0.553 0.739 0.944 0.948
0.9289 0.926 0.9919 1.0441 1.1113 026_884_00308160 Lung: NSCLC 884
RERF-LC-MS 456 3.239905 0.585 0.663 0.662 0.6574 0.7423 0.886
0.9636 0.9621 0.9957 026_796_00295871 Lung: NSCLC 796 NCI-H2009 456
3.244878 0.582 0.62 0.631 0.635 0.7036 0.747 0.8586 0.9443 0.9943
026_799_00295880 Lung: NSCLC 799 NCI-H661 456 3.295179 0.667 0.587
0.685 0.6862 0.8151 0.782 1.0055 0.9053 0.9863 026_822_00311150
Lung: NSCLC 822 NCI-H1869 456 3.570833 0.153 0.716 0.736 0.7399
0.8095 0.819 1.026 0.9674 1.0055 026_756_00302670 Lung: NSCLC 756
BEN 456 3.65632 0.639 0.833 0.942 0.9322 0.9128 0.965 0.9486 1.0116
1.0192 026_876_00299781 Lung: NSCLC 876 LU65 456 3.69068 0.635
0.695 0.704 0.7208 0.8211 0.878 0.9851 1.0211 1.0551
026_805_00304531 Lung: NSCLC 805 NCI-H1155 456 3.716538 0.61 0.788
0.787 0.8467 0.9573 0.91 0.953 1.0683 1.0508 026_834_00304610 Lung:
NSCLC 834 NCI-H2347 456 3.72347 0.653 0.736 0.756 0.8026 0.8524
0.984 0.9516 1.2251 1.0554 026_822_00304840 Lung: NSCLC 822
NCI-H1869 456 3.749478 0.594 0.736 0.791 0.7802 0.8592 0.824 0.8872
0.9703 1.0592 026_835_00302390 Lung: NSCLC 835 NCI-H2405 456
3.789193 0.693 0.659 0.651 0.6574 0.7337 0.768 0.8849 0.9046 1.0375
026_678_00304981 Lung: NSCLC 678 UMC-11 456 3.815948 0.655 0.751
0.848 0.8851 0.9411 1.013 0.9806 1.0842 1.1132 026_807_00314280
Lung: NSCLC 807 NCI-H650 456 3.884087 0.604 0.778 0.772 0.8577
0.8346 0.815 1.0068 0.831 0.9772 026_871_00299771 Lung: NSCLC 871
LK-2 456 3.898613 0.684 0.806 0.903 0.9071 0.958 1.018 0.9981
1.0604 1.0786 026_1249_00308150 Lung: NSCLC 1249 NCI-H720 456
3.907633 0.632 0.829 0.932 1.0847 1.526 0.799 0.8751 1.2042 1.1215
026_8231_00304510 Lung: NSCLC 8231 EMC-BAC-1 456 3.943946 0.675
0.713 0.751 0.8781 0.831 0.931 0.976 1.0569 1.0459 026_820_00304560
Lung: NSCLC 820 NCI-H1755 456 3.964037 0.658 0.769 0.785 0.8744
0.8799 0.931 0.9563 1.1261 1.0206 026_815_00304830 Lung: NSCLC 815
NCI-H1623 456 3.981628 0.646 0.709 0.712 0.7108 0.8124 0.879 0.7844
1.0245 1.0786 026_839_00302410 Lung: NSCLC 839 SW 900 456 3.982367
0.65 0.725 0.782 0.8242 0.8576 0.897 0.9535 1.0086 0.9978
026_804_00308480 Lung: NSCLC 804 NCI-H810 456 4.03318 0.731 0.763
0.91 0.9442 0.9087 1.003 0.9482 0.9716 1.0139 026_678_00309011
Lung: NSCLC 678 UMC-11 456 4.0744 0.514 0.846 0.86 0.931 0.9037
0.925 0.9312 0.9354 1.0786 026_842_00298540 Lung: NSCLC 842
NCI-H520 456 4.082969 0.729 0.811 0.916 1.0516 1.0142 1.002 1.0457
1.0025 0.9761 026_824_00314260 Lung: NSCLC 824 NCI-H1944 456
4.107799 0.695 0.731 0.691 0.7704 0.893 0.886 0.905 0.9791 1.0866
026_888_00298370 Lung: NSCLC 888 ABC-1 456 4.123019 0.64 0.676
0.717 0.7343 0.8328 0.84 0.8774 0.9195 0.9448 026_823_00298430
Lung: NSCLC 823 NCI-H1915 456 4.260241 0.691 0.707 0.702 0.7393
0.7957 0.834 0.9053 0.9612 0.9902 026_8232_00304520 Lung: NSCLC
8232 EMC-BAC-2 456 4.327709 0.712 0.795 0.825 0.8558 0.8815 0.959
0.9014 1.1842 1.0135 026_755_00300611 Lung: NSCLC 755 NCI-H1975 456
4.450033 0.666 0.968 0.963 0.9052 0.9346 0.928 0.9969 0.9681 0.973
026_868_00295440 Lung: NSCLC 868 PC-14 456 4.458594 0.726 0.893
0.842 0.8971 0.9361 1.057 0.9146 1.1281 0.933 026_872_00299750
Lung: NSCLC 872 HARA 456 4.480998 0.731 0.742 0.798 0.8152 0.8972
0.941 0.9735 1.0074 1.0301 026_800_00298441 Lung: NSCLC 800 NCI-H23
456 4.511394 0.702 0.698 0.786 0.7938 0.8549 0.889 0.9216 0.9447
1.013 026_836_00304620 Lung: NSCLC 836 NCI-H2444 456 4.530511 0.684
0.798 0.819 0.8507 0.863 0.86 0.9116 1.0106 1.0518 026_865_00296401
Lung: NSCLC 865 COR-L23 456 4.577187 0.791 0.78 0.815 0.8605 0.9148
0.924 1.1179 1.1287 1.1293 026_858_00300591 Lung: NSCLC 858 HCC-78
456 4.583505 0.703 0.983 0.936 1.06 0.9813 0.96 1.0028 0.9854
1.0584 026_854_00300681 Lung: NSCLC 854 EPLC-272H 456 4.590654
0.736 0.914 0.943 1.0019 1.0017 0.864 0.8937 0.9686 1.0373
026_837_00311160 Lung: NSCLC 837 NCI-H2122 456 4.595192 0.771 0.917
0.969 0.9878 0.9667 1.017 1.0165 1.0062 1.0324 026_8111_00308110
Lung: NSCLC 8111 LXF-289 456 4.689278 0.711 0.796 0.884 0.8648
0.868 0.89 0.9108 0.8979 0.9642 026_1136_00308471 Lung: NSCLC 1136
NCI-H1993 456 4.705234 0.725 0.768 0.682 0.7469 0.7866 0.883 0.9296
0.914 0.9652 026_827_00308860 Lung: NSCLC 827 NCI-H2085 456
4.711775 0.79 0.823 0.87 0.9311 0.924 1.007 0.967 1.0124 1.1093
026_859_00311090 Lung: NSCLC 859 HCC-827 456 4.735253 0.737 0.86
0.852 0.7863 0.8298 0.853 1.0366 1.1949 0.9783 026_886_00296261
Lung: NSCLC 886 EBC-1 456 4.744821 0.761 0.848 0.884 0.9053 0.9307
0.969 0.9903 0.9432 1.0947 026_8132_00308130 Lung: NSCLC 8132
NCI-H2126 456 4.781635 0.749 0.796 0.839 0.8331 0.8916 0.939 0.9049
0.9661 0.9286 026_793_00299711 Lung: NSCLC 793 NCI-H1781 456
4.904802 0.775 0.801 0.686 0.7429 0.7804 0.897 0.9136 0.9608 1.0112
026_1247_00314271 Lung: NSCLC 1247 NCI-H2291 456 4.945975 0.773
0.893 0.836 0.831 0.851 1.029 1.0615 0.9828 1.0493 026_860_00298400
Lung: NSCLC 860 LCLC-103H 456 4.97209 0.751 0.819 0.803 0.8985
0.8725 0.898 0.9606 0.9519 1.061 026_806_00300270 Lung: NSCLC 806
NCI-H647 456 5.044917 0.792 0.845 0.882 0.8698 0.8856 0.881 1.0769
0.9333 1.0648 026_877_00300630 Lung: NSCLC 877 PC-3 [JPC-3] 456
5.056812 0.847 0.832 0.968 0.9943 1.0985 1.01 1.0152 1.0063 1.1608
026_753_00298460 Lung: NSCLC 753 NCI-H460 456 5.061076 0.839 0.888
0.945 0.9015 1.041 1.039 1.021 0.9905 1.0126 026_844_00295461 Lung:
NSCLC 844 SW 1573 456 5.077017 0.798 0.834 0.833 0.8824 0.8946
0.971 1.0223 0.9943 1.0074 026_8088_00314320 Lung: NSCLC 8088
KNS-62 456 5.130485 0.86 0.847 0.879 0.9874 0.9738 1.047 1.0602
1.0303 0.9876 026_848_00300641 Lung: NSCLC 848 SK-LU-1 456 5.2771
0.811 0.91 0.946 0.9174 0.9429 0.922 0.9544 1.0372 1.0115
026_864_00304961 Lung: NSCLC 864 COR-L 105 456 5.319498 0.718 0.78
0.762 0.8018 0.7683 0.837 0.9721 0.8644 0.9527 026_677_00298361
Lung: NSCLC 677 A549 456 5.414187 0.763 0.799 0.843 0.8595 0.9001
0.919 0.9405 0.9783 0.996 026_8207_00304810 Lung: NSCLC 8207 LC-1F
456 5.423228 0.825 0.794 0.845 0.8457 0.8483 0.976 0.8421 0.8626
0.9602 026_833_00299790 Lung: NSCLC 833 NCI-H2342 456 5.548934
0.856 0.888 0.917 0.9241 0.9293 1.015 0.9449 1.0616 0.9624
026_870_00299801 Lung: NSCLC 870 RERF-LC-KJ 456 5.685511 0.892 0.96
0.999 0.9623 1.0469 1.054 1.0305 1.0128 1.0476 026_845_00311341
Lung: NSCLC 845 NCI-H1838 456 5.711103 0.847 0.867 0.909 0.9088
0.9208 0.932 0.9775 0.9857 0.9755
026_8103_00306210 Lung: NSCLC 8103 LC-2-ad 456 5.718708 0.836 0.96
1.004 0.9427 1.0257 1.064 1.0682 0.9834 1.007 026_794_00300601
Lung: NSCLC 794 NCI-H1792 456 5.721238 0.339 0.904 0.992 0.9248
0.9615 1.005 0.9405 0.9595 1.0225 026_890_00300220 Lung: NSCLC 890
H3255 456 5.722728 0.903 0.862 0.844 0.9926 1.005 0.998 1.1503
1.1306 1.1501 026_862_00304820 Lung: NSCLC 862 LOU-NH91 456
5.738178 0.886 0.864 0.837 0.9386 0.9258 0.887 0.948 1.0561 1.1752
026_812_00298410 Lung: NSCLC 812 NCI-H1437 456 5.791603 0.894 0.946
0.993 0.9231 0.9704 1.003 1.0591 1.0305 1.0361 026_862_00316560
Lung: NSCLC 862 LOU-NH91 456 5.867157 0.871 0.905 0.923 0.9415
0.9143 0.955 0.9491 0.9721 0.9923 026_857_00296270 Lung: NSCLC 857
HCC-44 456 5.930045 0.904 0.868 0.893 0.9249 0.9348 0.956 1.0259
0.9727 1.0027 026_679_00296240 Lung: NSCLC 679 ChaGo-K-1 456
5.965866 0.878 0.915 0.949 0.8968 0.942 0.953 0.9732 0.9711 1.0338
026_791_00298421 Lung: NSCLC 791 NCI-H1650 456 5.971987 0.888 0.896
0.979 0.9429 0.9543 0.921 0.9516 0.9551 0.9966 026_864_00311551
Lung: NSCLC 864 COR-L 105 456 5.975297 0.765 0.838 0.843 0.8384
0.8225 0.848 0.8917 0.8749 0.9811 026_846_00300620 Lung: NSCLC 846
NCI-H2030 456 6.014339 0.886 0.979 0.985 0.9316 1.0428 0.925 0.9778
1.056 0.9904 026_841_00296291 Lung: NSCLC 841 NCI-H2170 456 6.01763
0.897 0.938 0.928 0.9356 0.9557 0.955 1.0968 1.0708 1.0963
026_813_00308231 Lung: NSCLC 813 NCI-H1563 456 6.06364 0.856 0.979
0.977 0.9369 0.9242 0.912 0.8859 0.9059 0.8856 026_816_00295860
Lung: NSCLC 816 NCI-H1651 456 6.160326 0.864 1.039 1.026 1.008
1.0776 0.871 1.0553 1.0199 0.9978 026_818_00308241 Lung: NSCLC 818
NCI-H1703 456 6.172519 0.946 0.829 0.858 0.9042 0.9197 0.887 0.9097
0.9729 0.9634 026_808_00296300 Lung: NSCLC 808 NCI-H838 456
6.172929 0.922 0.941 0.955 1.022 0.964 1.043 1.0358 1.0134 1.0658
026_855_00302330 Lung: NSCLC 855 HCC-15 456 6.195295 0.868 0.911
0.901 0.8895 0.9071 0.95 0.9608 1.0091 1.0383 026_803_00298470
Lung: NSCLC 803 NCI-H522 456 6.200929 0.878 0.933 0.953 0.8695
0.9015 0.833 0.8882 0.8789 1.1066 026_832_00303110 Lung: NSCLC 832
NCI-H2228 456 6.240586 0.908 0.872 0.907 0.9005 0.9724 0.974 0.9776
0.9706 0.968 026_874_00308171 Lung: NSCLC 874 RERF-LC-Sql 456
6.282697 1.064 0.854 0.829 0.8009 0.8914 0.943 0.8612 0.9931 0.9393
026_879_00308101 Lung: NSCLC 879 LU99A 456 6.327465 0.881 0.987
0.995 1.0199 0.9725 0.948 0.9588 0.9756 0.9692 026_856_00299760
Lung: NSCLC 856 HCC-366 456 6.328052 0.936 0.943 0.959 1.0286
0.9991 1.002 1.0072 1.0077 1.0015 026_798_00304790 Lung: NSCLC 798
Calu-3 456 6.338698 0.926 0.986 0.869 0.9635 1.0683 1.166 1.1053
1.1123 1.1003 026_825_00308250 Lung: NSCLC 825 NCI-H2023 456
6.435057 0.938 0.885 0.929 0.961 0.9166 0.959 0.9502 0.9835 0.9846
026_8072_00306200 Lung: NSCLC 8072 HOP-62 456 6.481774 0.973 0.958
0.931 0.9405 0.9799 0.901 1.0374 0.9717 1.1512 026_843_00296331
Lung: NSCLC 843 SK-MES-1 456 6.499026 0.91 0.99 0.934 0.9509 0.9694
1.003 0.9804 0.9674 1.0295 026_1251_00318710 Lung: NSCLC 1251
NCI-H835 456 6.684349 0.964 1.054 0.854 0.9853 0.77 0.963 0.8714
0.9637 1.1416 026_816_00302370 Lung: NSCLC 816 NCI-H1651 456
6.769433 0.903 1.072 1.021 1.002 1.0583 0.94 1.0664 1.0462 1.0774
026_801_00295941 Lung: NSCLC 801 NCI-H1299 456 6.808075 1.036 0.898
0.912 0.9194 0.9832 0.955 1.0252 0.9946 1.085 026_850_00311050
Lung: NSCLC 850 201T 456 6.874432 0.942 0.957 0.988 0.989 0.9596
0.985 0.9759 0.9839 0.9726 026_818_00309001 Lung: NSCLC 818
NCI-H1703 456 7.192409 0.889 0.857 0.855 0.8355 0.8207 0.856 0.8191
0.9124 0.9468 026_1247_00304851 Lung: NSCLC 1247 NCI-H2291 456
7.252153 1.162 1.047 0.877 1.034 0.8759 0.694 1.2149 1.0391 1.1021
026_819_00306250 Lung: NSCLC 819 NCI-H1734 456 7.342705 1.046 1.007
1.012 1.018 0.9902 0.945 0.9742 0.9718 1.0221 026_821_00306261
Lung: NSCLC 821 NCI-H1793 456 7.345651 1.015 1.1 1.058 1.0171
1.0121 1.065 1.0342 1.0776 1.0468 026_798_00308080 Lung: NSCLC 798
Calu-3 456 7.389624 1.032 1 0.914 0.9927 1.0342 1.004 1.0095 0.9327
0.938 026_678_00306181 Lung: NSCLC 678 UMC-11 456 7.458364 1.03
1.058 1.054 1.0958 1.0021 1.03 1.0784 1.0779 1.0688
026_797_00308730 Lung: NSCLC 797 NCI-H596 456 7.556662 1.124 1.076
0.92 0.9992 0.9285 0.936 0.911 1.0276 0.8862 026_790_00306231 Lung:
NSCLC 790 NCI-H1573 456 7.606756 1.049 1 1.054 1.0146 1.0291 0.99
1.0304 1.0221 1.0512 026_752_00306191 Lung: NSCLC 752 A-427 456
7.608444 1.109 0.988 1.054 1.0224 0.9429 1.084 1.0956 1.0273 1.0528
026_845_00306271 Lung: NSCLC 845 NCI-H1838 456 7.666847 1.074 1.08
1.064 1.0779 1.0805 1.046 1.0674 0.9941 1.02 026_8133_00306280
Lung: NSCLC 8133 NCI-H322M 456 7.713112 1.158 1.06 1.135 1.1499
1.1305 1.103 1.1464 1.1475 1.0913 026_8130_00306220 Lung: NSCLC
8130 NCI-H1355 456 7.765888 1.12 1.188 1.002 0.9001 0.8103 1.095
0.9608 0.9714 0.9997 026_683_00306241 Lung: NSCLC 683 NCI-H1581 456
7.774034 1.163 1.024 0.949 1.0959 1.0605 1.114 1.0118 0.9537 0.9212
026_792_00306140 Lung: NSCLC 792 NCI-H1666 456 7.825548 1.136 1.069
1.132 1.0901 1.0688 1.097 1.0684 1.0463 1.1115 026_840_00306151
Lung: NSCLC 840 NCI-H441 456 7.946249 1.088 1.059 1.052 1.0622
1.0476 1.049 1.0439 1.045 1.0714 026_8075_00306131 Lung: NSCLC 8075
IA-LM 456 7.984543 1.046 1.042 1.043 1.0424 1.0323 1.048 1.0441
1.0279 1.0694 026_61_00285570 Lymphoma 61 JSC-1 456 0.316646 0.313
0.329 0.341 0.3908 0.5654 0.63 0.724 0.8346 0.8527
026_8222_00291350 Lymphoma 8222 H9 456 0.402947 0.102 0.284 0.272
0.3996 0.5762 0.732 0.8412 0.8346 0.8529 026_140_00291320 Lymphoma
140 A3/KAW 456 0.717935 0.185 0.296 0.408 0.4762 0.5561 0.736
0.8297 0.8761 0.8425 026_237_00291380 Lymphoma 237 SU-DHL-16 456
0.982467 0.155 0.278 0.405 0.5737 0.6759 0.816 0.9129 0.9363 0.9318
026_220_00288720 Lymphoma 220 OCI-LY-19 456 1.28484 0.324 0.393
0.455 0.5338 0.6443 0.826 0.9343 0.9969 1.073 026_257_00285640
Lymphoma 257 WIL2 NS 456 1.523653 0.32 0.456 0.509 0.6199 0.6926
0.771 0.8518 0.9524 0.934 026_239_00288750 Lymphoma 239 SU-DHL-5
456 1.616577 0.174 0.577 0.461 0.7856 0.7368 0.845 0.9512 0.9535
1.2005 026_124_00287850 Lymphoma 124 BC-1 456 1.816766 0.239 0.53
0.6 0.6778 0.8612 0.946 0.9754 0.9846 1.0038 026_104_00287960
Lymphoma 104 TUR 456 2.217781 0.396 0.51 0.597 0.7468 0.9062 0.94
0.9822 0.9761 1.0165 026_8199_00291330 Lymphoma 8199 CTB-1 456
2.341488 0.445 0.509 0.654 0.6993 0.7792 0.767 0.8214 0.8543 0.9676
026_69_00283480 Lymphoma 69 CA46 456 2.43077 0.358 0.6 0.775 0.78
0.8817 0.896 0.9912 0.9446 0.9358 026_112_00285620 Lymphoma 112 SR
456 2.466522 0.224 0.573 0.653 0.7668 0.8482 0.866 0.9087 0.9409
0.9255 026_241_00288760 Lymphoma 241 SU-DHL-8 456 2.509783 0.398
0.583 0.743 0.8249 0.935 0.956 1.0439 0.9903 1.0527
026_255_00285610 Lymphoma 255 Sci-1 456 2.705483 0.445 0.61 0.788
0.9473 0.9153 1.003 0.9035 0.8642 0.8757 026_62_00293930 Lymphoma
62 IM-9 456 2.809638 0.572 0.562 0.594 0.6839 0.7773 1.18 0.876
1.0089 1.0329 026_93_00287880 Lymphoma 93 HH 456 2.828886 0.523
0.575 0.731 0.804 0.9283 0.94 8 0.9965 0.999 1.0069
026_216_00290710 Lymphoma 216 NU-DUL-1 456 2.880782 0.459 0.664
0.767 0.8093 0.7784 0.884 0.9338 0.9363 0.9519 026_123_00287860
Lymphoma 123 BC-3 456 2.893681 0.495 0.678 0.776 0.9051 1.0108
1.003 1.0056 0.9841 0.9941 026_113_00288520 Lymphoma 113 DB 456
2.901204 0.479 0.705 0.868 0.9097 0.9559 0.984 1.0104 0.992 1.0316
026_8035_00303290 Lymphoma 8035 DOHH-2 456 2.927461 0.508 0.78
0.831 0.8009 0.9998 0.997 1.0295 0.9393 1.0203 026_240_00300290
Lymphoma 240 SU-DHL-6 456 2.933827 0.208 0.663 0.733 0.7473 0.7814
0.839 0.8554 0.9327 1.029 026_248_00285630 Lymphoma 248 VAL 456
2.964017 0.528 0.639 0.739 0.9112 0.9265 0.96 1.0023 0.9827 0.9859
026_162_00290790 Lymphoma 162 HDLM-2 456 2.980685 0.56 0.811 0.678
0.7341 0.7627 0.782 0.7915 0.9638 0.7387 026_105_00287940 Lymphoma
105 RPMI 6666 456 3.064381 0.516 0.701 0.691 0.7257 0.8234 0.904
0.9883 1.0013 1.033 026_240_00302920 Lymphoma 240 SU-DHL-6 456
3.114948 0.326 0.629 0.741 0.7802 0.8119 0.831 0.9347 0.921 0.9838
026_163_00287730 Lymphoma 163 HD-MY-Z 456 3.123344 0.582 0.606
0.654 0.6825 0.785 0.896 0.9395 0.98 1.0207 026_139_00283530
Lymphoma 139 MC116 456 3.12577 0.085 0.686 0.917 0.9565 0.9611 0.99
0.9875 0.9789 0.9559 026_133_00293941 Lymphoma 133 NK-92M1 456
3.297366 0.637 0.637 0.757 0.8911 0.8633 1.136 1.0548 0.89 0.9001
026_282_00287750 Lymphoma 282 P32/ISH 456 3.328203 0.416 0.744
0.875 0.8947 0.8754 0.907 0.9959 0.9144 0.9277 026_80_00287950
Lymphoma 80 ST486 456 3.520498 0.263 0.688 0.699 0.7969 0.8071
0.904 0.8902 0.9142 1.017 026_73_00283500 Lymphoma 73 EB-3 456
3.528971 0.566 0.739 0.806 0.8214 0.8547 1.09 0.8321 0.8619 0.8548
026_60_00287891 Lymphoma 60 JM1 456 3.618767 0.577 0.678 0.734
0.7932 0.8209 0.833 0.8444 0.8567 0.902 026_185_00288820 Lymphoma
185 L-540 456 3.666174 0.626 0.88 0.936 1.0302 1.0245 1.037 1.0524
1.0175 1.0525 026_70_00283490 Lymphoma 70 Daudi 456 3.687192 0.603
0.789 0.925 0.8867 0.9929 0.987 0.8586 0.9555 0.8243
026_228_00291370 Lymphoma 228 RC-K8 456 3.710403 0.605 0.678 0.732
0.7776 0.8387 0.943 0.9437 0.9368 0.9149 026_74_00285600 Lymphoma
74 Raji 456 3.736373 0.639 0.768 0.89 0.8258 0.9671 0.97 0.9444
0.9665 1.046 026_173_00288920 Lymphoma 173 KARPAS-422 456 3.812903
0.638 0.787 0.876 0.8937 0.8652 0.959 0.9976 0.9896 1.005
026_125_00287910 Lymphoma 125 MC/CAR 456 3.860302 0.687 0.792 0.873
0.8871 1.009 1.011 0.9976 0.987 1.0162 026_280_00285420 Lymphoma
280 SCC-3 456 3.903427 0.648 0.86 0.895 0.9301 0.9335 0.934 1.0707
0.9 0.9445 026_242_00288770 Lymphoma 242 SUP-HD1 456 4.023177 0.671
0.815 0.875 0.8573 0.8914 0.965 1.0137 0.9754 1.1043
026_160_00285410 Lymphoma 160 GRANTA-519 456 4.112789 0.631 0.706
0.761 0.7798 0.7991 0.771 0.9167 0.9366 0.9971 026_144_00287710
Lymphoma 144 BL-41 456 4.441898 0.536 1.076 0.941 0.9045 0.8922
0.893 0.9542 0.9696 1.0289 026_184_00290800 Lymphoma 184 L-428 456
4.489131 0.614 0.705 0.723 0.7315 0.7351 0.721 1.0011 0.7501 0.8687
026_128_00290690 Lymphoma 128 Farage 456 4.585156 0.766 0.834 0.861
0.8151 0.9324 0.908 1.3723 1.1697 1.2229 026_250_00285660 Lymphoma
250 WSU-NHL 456 4.693667 0.719 0.765 0.866 0.8242 0.9178 0.947
0.9725 0.9585 0.9714 026_182_00290700 Lymphoma 182 L-1236 456
4.714251 0.76 0.859 0.875 0.8522 0.9702 0.962 1.0483 0.9432 1.3018
026_95_00283510 Lymphoma 95 HT 456 4.721481 0.719 0.842 0.88 0.8266
0.8534 0.902 1.0348 0.9564 0.966 026_264_00288840 Lymphoma 264 TK
456 4.750226 0.794 0.966 0.936 0.9709 1.0212 1.024 1.0348 1.0318
1.0681 026_266_00287760 Lymphoma 266 SLVL 456 4.90866 0.731 0.786
0.794 0.8012 0.8742 0.873 0.9435 0.9199 0.977 026_75_00282830
Lymphoma 75 Jiyoye 456 4.937844 0.801 0.99 0.974 1.0523 1.196 1.107
1.0641 1.1044 1.0749 026_111_00291360 Lymphoma 111 Hs 445 456
5.022248 0.704 0.734 0.703 0.794 0.7451 0.779 0.8243 0.8675 0.946
026_8151_00288540 Lymphoma 8151 Ramos-2G6-4C10 456 5.147746 0.242
0.868 0.954 0.9827 1.022 0.974 0.9967 0.9674 1.0529
026_156_00287720 Lymphoma 156 DG-75 456 5.222394 0.834 1.001 1.03
1.0349 0.9848 0.996 1.012 0.9965 0.9857 026_243_00296620 Lymphoma
243 SUP-M2 456 5.25553 0.648 0.88 0.96 0.9351 0.9518 0.968 0.9742
0.9627 0.9659 026_86_00287870 Lymphoma 86 EB2 456 5.287284 0.86
1.017 1.01 1.0356 1.0217 1.05 1.0267 1.0118 1.0227 026_162_00288790
Lymphoma 162 HDLM-2 456 5.425219 0.877 0.903 0.797 0.9228 1.0293
1.043 1.0157 1.0517 1.0837 026_172_00287740 Lymphoma 172 KARPAS-299
456 5.433867 0.832 0.891 0.95 0.9494 0.955 0.954 0.9499 0.9633
0.9487 026_235_00285430 Lymphoma 235 SU-DHL-1 456 6.401484 0.885
0.997 0.987 0.9424 0.9387 0.884 0.8642 0.9464 0.9439
026_143_00288690 Lymphoma 143 AMO-1 456 6.692908 0.924 1.02 5 1.129
1.034 1.1112 1.034 1.0278 1.0136 1.0255 026_193_00288830 Lymphoma
193 MHH-PREB-1 456 6.799634 0.622 0.958 1.012 1.0149 1.0368 1.025
1.0064 0.9803 0.989 026_236_00288730 Lymphoma 236 SU-DHL-10 456
6.99662 0.175 0.993 1.05 1.0763 1.0476 1.029 1.0509 1.0397 1.0494
026_81_00288530 Lymphoma 81 GA-10 456 7.031749 0.608 1.003 1.009
1.1031 1.0252 1.021 1.0113 1.0077 1.0848 026_251_00287840 Lymphoma
251 YT 456 7.411746 0.64 0.986 1.008 1.0201 1.0204 1.022 1.0267
0.9848 1.0079 026_178_00288810 Lymphoma 178 KM-H2 456 7.739504
1.008 1.009 0.976 1.0197 1.0404 1.036 1.0412 1.0156 1.0276
026_249_00285650 Lymphoma 249 WSU-DLCL2 456 8.212885 1.047 1.226
1.172 1.1142 1.0869 1.071 0.9837 1.0247 1.0207 026_238_00288740
Lymphoma 238 SU-DHL-4 456 8.647656 1.061 1.077 1.045 1.0257 1.0384
1.029 1.0236 1.018 1.0204 026_131_00287930 Lymphoma 131 RL 456
8.679308 1.124 1.094 1.048 1.024 1.0293 1.012 1.0293 0.9824 1.0061
026_915_00269070 Miscellaneous 915 Hs 633T 456 1.797943 0.344 0.532
0.607 0.6061 0.6357 0.736 0.8936 1.0346 1.1375 026_911_00269060
Miscellaneous 911 GCT 456 2.424125 0.433 0.617 0.642 0.6508 0.655
0.804 0.7763 0.9066 0.9967 026_8172_00269460_ Miscellaneous 8172
SW872 456 3.196748 0.511 0.716 0.719 0.7459 0.8264 0.881 0.9107
0.9313 1.0224 026_913_00271970 Miscellaneous 913 JAR 456 3.876744
0.685 0.695 0.794 0.851 0.9936 1.011 0.9013 1.0881 1.0491
026_8194_00271980 Miscellaneous 8194 JEG-3 456 3.901223 0.623 0.808
0.876 0.9097 0.9037 0.874 0.9014 0.943 1.1354 026_8171_00269120
Miscellaneous 8171 SW684 456 3.978538 0.693 0.91 0.98 1.1114 1.0275
1.032 0.9881 0.9218 1.0725 026_8112_00269450 Miscellaneous 8112
MFH-ino 456 4.013788 0.691 0.674 0.795 0.8068 0.8378 0.91 0.9824
1.0389 1.0625 026_916_00271320 Miscellaneous 916 HT 1080 456
4.254215 0.695 0.804 0.766 0.7943 0.8276 0.873 1.1135 1.1006 1.1093
026_8004_00271280 Miscellaneous 8004 A388 456 4.494172 0.743 0.793
0.841 0.8441 0.8899 0.888 1.1408 0.925 1.1327 026_8112_00271340
Miscellaneous 8112 MFH-ino 456 4.624376 0.721 0.732 0.741 0.7501
0.7753 0.799 1.017 0.9044 1.0866 026_8004_00269400 Miscellaneous
8004 A388 456 4.778414 0.747 0.764 0.776 0.8164 0.8628 0.861 0.9217
0.8924 1.0697 026_8192_00269470 Miscellaneous 8192 VA-ES-BJ 456
4.956405 0.78 0.867 0.874 0.984 0.9192 0.889 0.9737 1.0227 1.0186
026_8194_00269440 Miscellaneous 8194 JEG-3 456 5.058221 0.786 0.87
1.048
0.908 0.8876 0.984 0.8804 1.0294 1.0609 026_913_00269430
Miscellaneous 913 JAR 456 5.170211 0.816 0.841 0.849 0.8709 0.8661
0.88 1.0502 1.0585 1.0591 026_916_00269420 Miscellaneous 916 HT
1080 456 5.302771 0.831 0.891 0.904 0.9053 0.9327 0.978 0.9465
1.1023 1.1108 026_8175_00269140 Miscellaneous 8175 SW982 456
5.586364 0.832 0.968 0.988 1.1946 1.0638 1.017 1.0728 1.1006 1.2781
026_1225_00269660 Muscle 1225 RD 456 2.694207 0.612 0.595 0.602
0.575 0.6051 0.678 0.8404 0.9183 0.947 26_135_00271680 Muscle 135
SJCRH30 456 2.740947 0.247 0.61 0.713 0.8654 0.8633 0.854 0.9433
0.9991 1.105 026_924_00269640 Muscle 924 A673 456 2.779569 0.563
0.554 0.607 0.6884 0.7085 0.808 0.8122 0.8796 0.9437
26_562_00271660 Muscle 562 KYM-1 456 2.818237 0.433 0.665 0.727
0.7663 0.7943 0.835 0.9132 0.9948 0.9596 026_135_00271410 Muscle
135 SJCRH30 456 3.059385 0.345 0.672 0.79 0.8395 0.8741 0.957
0.9768 1.032 1.1796 026_923_00269680 Muscle 923 RH-41 456 3.133389
0.309 0.65 0.731 0.7895 0.6971 0.795 0.7303 0.9458 0.9325
026_562_00270060 Muscle 562 KYM-1 456 3.396905 0.586 0.748 0.78
0.8707 0.8698 0.955 1.0372 1.0323 1.0217 026_920_00269670 Muscle
920 RH-1 456 3.433085 0.581 0.772 0.846 0.9032 0.9238 0.951 0.9496
0.9615 0.9852 026_135_00270070 Muscle 135 SJCRH30 456 3.759361
0.326 0.727 0.825 0.8737 0.9084 0.931 0.9701 0.9946 0.9435
026_919_00269630 Muscle 919 A-204 456 5.25605 0.778 0.798 0.868
0.8744 0.861 0.89 0.9735 0.9392 0.9765 026_921_00285151 Muscle 921
RH-18 456 5.863422 0.84 0.987 0.961 0.9653 0.9811 0.903 0.8875
0.9228 0.9815 026_8182_00293791 Muscle 8182 TE-441-T 456 6.133177
0.848 1.001 1.103 1.0954 1.1007 0.855 0.8882 1.0698 0.8069
026_369_00258500 Nervous System 369 CHP-212 456 -4.151196 0.165
0.184 0.183 0.1873 0.1985 0.219 0.2948 0.3285 0.4946
026_390_00262920 Nervous System 390 NB69 456 -1.3827 0.056 0.074
0.068 0.0948 0.096 0.163 0.4343 0.6938 0.8833 026_629_00316570
Nervous System 629 NB(TU)1-10 456 -0.654294 0.322 0.361 0.388 0.398
0.4347 0.594 0.7533 0.6321 0.6782 026_366_00257090 Nervous System
366 BE(2)-M17 456 -0.258429 0.204 0.352 0.333 0.3872 0.4128 0.547
0.6415 0.7687 0.9298 026_384_00314240 Nervous System 384 MHH-NB-11
456 1.316182 0.368 0.42 0.48 0.6318 0.5903 0.783 0.7802 0.9243
0.5317 026_385_00314290 Nervous System 385 SIMA 456 1.359572 0.402
0.496 0.488 0.5135 0.6006 0.629 0.8787 0.8808 1.0864
026_630_00264810 Nervous System 630 NH-12 456 1.516245 0.143 0.443
0.537 0.5763 0.6816 0.684 0.7263 0.9504 0.791 026_8124_00308720
Nervous System 8124 NB14 456 1.784337 0.802 0.401 0.6 0.6379 0.6871
0.943 0.8827 0.8247 0.9343 026_639_00271120 Nervous System 639 GOTO
456 1.784838 0.174 0.52 0.54 0.6353 0.6485 0.726 0.9341 0.8908
1.1295 026_8094_00314230 Nervous System 8094 LAN-6 456 1.824343
0.55 0.565 0.476 0.5734 0.6862 0.656 0.7893 0.9264 0.7016
026_8076_00260401 Nervous System 8076 IMR-5 456 1.864038 0.346
0.489 0.575 1.2246 0.8619 0.823 1.0781 0.7607 0.8894
026_8127_00258950 Nervous System 8127 NB5 456 2.106125 0.202 0.628
0.611 0.6823 0.725 0.845 0.9541 0.9703 1.0012 026_8124_00311130
Nervous System 8124 NB14 456 2.216808 0.544 0.563 0.516 0.5711
0.5857 0.776 0.9014 0.9091 1.0395 026_8126_00264800 Nervous System
8126 NB17 456 2.289546 0.347 0.54 0.684 0.7567 0.8851 0.929 0.8729
0.9752 0.9196 026_641_00308180 Nervous System 641 TGW 456 2.326645
0.456 0.502 0.593 0.7234 0.8006 0.88 0.9296 0.9651 0.9821
026_8121_00256290 Nervous System 8121 NB10 456 2.356578 0.646 0.568
0.562 0.5835 0.6642 0.613 0.5487 0.6035 1.0005 026_8220_00308810
Nervous System 8220 KP-N-YN 456 2.470465 0.366 0.619 0.752 0.7815
0.7642 0.87 0.8457 0.8335 0.8942 026_8090_00263920 Nervous System
8090 KP-N-YS 456 2.547617 0.267 0.583 0.69 0.9135 0.905 0.924
0.9505 0.9232 1.012 026_363_00252730 Nervous System 363 SK-N-FI 456
2.667265 0.519 0.583 0.574 0.6332 0.5059 0.571 0.7766 0.6898 0.9188
026_382_00311110 Nervous System 382 KELLY 456 3.39659 0.518 0.757
0.767 0.7123 0.7538 0.883 0.8718 0.8733 0.9755 026_8064_00259120
Nervous System 8064 GI-ME-N 456 3.426922 0.582 0.763 0.78 0.8295
0.8371 0.955 1.0539 1.0652 1.104 026_8195_00318640 Nervous System
8195 CHP-134 456 3.519288 0.6 0.8 0.86 0.909 0.9931 0.919 0.9533
0.941 1.0273 026_382_00308690 Nervous System 382 KELLY 456 3.678148
0.59 0.706 0.666 0.7413 0.8277 0.814 0.8335 0.8742 0.9843
026_8129_00314250 Nervous System 8129 NB7 456 3.824042 0.644 0.861
0.945 0.972 0.9655 0.998 1.0941 1.0771 1.0471 026_8126_00280241
Nervous System 8126 NB17 456 3.845261 0.611 0.771 0.746 1.0569
0.8567 0.864 0.9408 0.9041 0.972 026_8007_00252850 Nervous System
8007 ACN 456 3.989797 0.411 0.72 0.821 0.7359 0.7717 0.797 0.8528
1.0702 0.9564 026_8122_00273460 Nervous System 8122 NB12 456
4.358809 0.15 0.809 0.911 0.9912 0.9113 1.021 0.9055 0.8723 1.0083
026_8226_00252530 Nervous System 8226 NBsusSR 456 4.365592 1.14
0.81 0.878 0.6567 0.6737 0.94 0.7615 0.8399 1.1136
026_8064_00256670 Nervous System 8064 GI-ME-N 456 4.809816 0.398
0.871 0.839 0.9402 0.8757 0.96 0.9284 0.9838 1.0451
026_8128_00257200 Nervous System 8128 NB6 456 4.886145 0.836 0.93
0.956 1.0121 0.9926 1.008 0.9893 0.9944 0.9472 026_8123_00256300
Nervous System 8123 NB13 456 5.775254 0.837 1.028 1.034 0.9756
1.0272 0.974 0.9893 0.9781 0.9486 026_396_00261020 Nervous System
396 NB-1 456 5.873385 0.813 1.039 0.892 0.8527 0.9085 0.934 0.9583
0.9709 1.0441 026_370_00252740 Nervous System 370 SK-N-SH 456
6.176056 0.926 0.811 0.891 1.0803 2.0427 1.01 0.892 0.9269 0.9864
026_364_00252720 Nervous System 364 SK-N-DZ 456 6.251974 0.895
1.012 0.958 1.0963 0.9642 0.976 1.0005 0.9787 0.9774
026_370_00258530 Nervous System 370 SK-N-SH 456 6.266144 0.36 0.933
1.052 1.0961 1.0754 0.948 0.9798 1.0134 0.9668 026_362_00252710
Nervous System 362 SK-N-AS 456 6.874316 0.911 1.075 0.916 1.0354
0.9179 0.939 1.178 0.9451 0.9362 026_368_00252700 Nervous System
368 MC-IXC 456 6.982443 0.084 0.616 1.087 1.1559 1.0678 1.11 1.0784
0.9917 0.9902 026_8122_00262490 Nervous System 8122 NB12 456
7.149853 0.063 0.973 1.019 1.3067 1.0528 0.999 1.4063 0.9955 0.9632
026_934_00287430 Ovary 934 A2780 456 -0.227264 0.302 0.3 0.342
0.3503 0.3758 0.467 0.7239 0.7489 0.9268 026_1126_00293760 Ovary
1126 OV-90 456 0.856076 0.323 0.361 0.384 0.4703 0.5785 0.701
0.8275 0.885 0.873 026_1129_00290660 Ovary 1129 TOV-112D 456
2.270257 0.47 0.481 0.592 0.7498 0.7692 0.925 0.9217 0.9942 1.1258
026_1220_00291150 Ovary 1220 ES-2 456 2.397868 0.584 0.504 0.584
0.5655 0.9851 0.957 1.1602 1.1706 1.0966 026_949_00290350 Ovary 949
TYK-nu 456 2.489488 0.511 0.577 0.587 0.5768 0.6153 0.686 0.7718
1.0687 1.0378 026_925_00290850 Ovary 925 IGROV-1 456 2.753994 0.494
0.608 0.578 0.7594 0.7462 0.857 0.9169 0.8966 0.8943
026_8244_00314220 Ovary 8244 JHOS-4 456 2.779507 0.63 0.601 0.597
0.6381 0.7711 0.88 0.955 0.9128 0.9028 026_8279_00293400 Ovary 8279
UWB1.289 456 2.893774 0.764 0.625 0.723 0.7182 0.783 1.079 0.9597
1.044 1.0467 026_940_00290340 Ovary 940 RMG-I 456 3.11591 0.587
0.604 0.66 0.6995 0.6871 0.846 0.8531 0.9424 1.0257
26_8238_00304370 Ovary 8238 IOSE-364- 456 3.149354 0.505 0.763
0.761 0.7451 0.8102 0.848 1.0558 1.0226 1.0295 026_8237_00295840
Ovary 8237 Hey 456 3.317969 0.551 0.783 0.688 0.776 0.8484 0.917
1.1038 1.0681 1.1892 026_8260_00292720 Ovary 8260 PEO1 456 3.508406
0.648 0.61 0.642 0.7269 0.6891 0.74 1.0027 0.9408 0.9148
026_8230_00303100 Ovary 8230 DOV13 456 3.68994 0.636 0.73 0.886
0.851 0.9406 0.961 0.9388 0.9681 1.0998 026_8230_00292650 Ovary
8230 DOV13 456 3.809459 0.641 0.685 0.768 0.9488 0.8764 0.863
0.9076 0.9226 1.0565 026_8092_00295850 Ovary 8092 KURAMOCHI 456
3.819916 0.757 0.729 0.642 0.6589 0.7541 0.914 0.9211 1.0035 1.0531
026_8240_00291160 Ovary 8240 IOSE-523- 456 3.834885 0.676 0.747
0.827 0.9249 0.9868 1 0.9181 0.9742 1.0197 026_932_00291140 Ovary
932 EFO-27 456 3.943589 0.65 0.668 0.65 0.7216 0.7753 0.887 0.9104
0.9136 0.9152 026_8084_00292670 Ovary 8084 KGN 456 4.00968 0.776
0.686 0.68 0.804 0.816 0.93 0.911 0.9826 0.9092 026_1125_00290571
Ovary 1125 Caov-3 456 4.050855 0.676 0.787 0.855 0.951 0.9041 1.029
0.9092 0.8992 1.0107 026_8256_00292920 Ovary 8256 OV-7 456 4.064163
0.666 0.844 0.821 0.9099 0.9293 0.961 1.0098 0.9638 0.9379
026_8241_00292900 Ovary 8241 IOSE-75-16SV40 456 4.094216 0.698
0.804 0.834 0.8672 0.894 1.02 1.0443 1.0711 1.107 026_938_00287450
Ovary 938 OAW42 456 4.158942 0.689 0.707 0.728 0.8613 0.7453 0.912
0.7815 1.0258 1.0515 026_1235_00298501 Ovary 1235 Caov-4 456
4.19878 0.673 0.815 0.851 0.7794 0.8482 0.895 1.0046 1.0048 0.9449
026_1221_00293780 Ovary 1221 SW 626 456 4.261643 0.636 0.704 0.69
0.6894 0.7014 0.755 0.8172 0.8859 1.0281 026_933_00295510 Ovary 933
FU-OV-1 456 4.35795 0.921 0.786 0.66 0.785 0.7855 0.932 1.0797
1.0643 1.0563 026_8259_00292710 Ovary 8259 OVK-18 456 4.419332
0.714 0.849 0.926 0.7816 0.8328 1.11 1.0158 0.9241 1.1619
026_8242_00295400 Ovary 8242 JHOS-2 456 4.46038 0.744 0.845 0.843
0.8069 0.9687 1.09 0.9491 1.0683 1.0333 026_1128_00292620 Ovary
1128 PA-1 456 4.524728 0.741 0.875 0.809 0.8925 0.9409 1.005 1.0033
1.0111 1.0682 026_938_00290330 Ovary 938 OAW42 456 4.600882 0.772
0.772 0.843 0.9103 0.8456 1.001 1.0305 1.0314 1.0721
026_8148_00292700 Ovary 8148 OVCAR-4 456 4.649732 0.715 0.824 0.723
0.6704 0.8391 0.855 0.9034 1.0922 1.0169 026_8243_00295410 Ovary
8243 JHOS-3 456 4.699022 0.801 0.758 0.691 0.7427 0.8184 0.86
1.0104 0.9653 1.0478 026_1130_00308501 Ovary 1130 TOV-21G 456
4.743414 0.744 0.838 0.882 0.8952 0.9369 0.922 0.9212 0.9456 0.9683
026_931_00252940 Ovary 931 EFO-21 456 4.786052 0.749 0.962 0.957
1.0202 1.0143 0.983 0.9976 0.9688 0.9523 026_931_00290820 Ovary 931
EFO-21 456 4.852802 0.791 0.924 0.954 0.9827 1.0247 0.961 0.9353
0.9274 0.9237 026_932_00288450 Ovary 932 EFO-27 456 4.890908 0.759
0.729 0.783 0.7813 0.828 0.851 0.9362 1.0032 1.0515
026_8258_00291180 Ovary 8258 OVCA433 456 4.901177 0.703 0.713 0.753
0.7366 0.79 0.864 0.9468 0.9244 0.9669 026_928_00290870 Ovary 928
OVCAR-5 456 4.903025 0.791 0.718 0.699 0.6913 0.7147 0.8 0.831
0.8925 0.9149 026_8257_00293650 Ovary 8257 OVCA420 456 4.98883
0.716 0.738 0.79 0.8055 0.8238 0.819 0.8677 0.8812 1.0222
026_8257_00292690 Ovary 8257 OVCA420 456 5.050268 0.789 0.822 0.798
0.9932 0.8382 0.876 0.928 0.9213 1.1024 026_8239_00292890 Ovary
8239 IOSE-397 456 5.092878 0.842 0.8 0.866 0.8796 0.8926 0.921
1.2015 0.9976 1.2125 026_941_00288490 Ovary 941 RKN 456 5.095259
0.777 0.883 0.798 0.796 0.8682 1.049 1.195 1.0697 1.0119
026_929_00290880 Ovary 929 OVCAR-8 456 5.147098 0.763 0.805 0.822
0.8457 0.8675 0.888 0.9211 0.9687 1.0047 026_8215_00287660 Ovary
8215 OC-314 456 5.38803 0.869 0.99 0.986 1.0451 1.0092 1.01 1.0495
0.9963 1.0021 026_939_00287460 Ovary 939 SK-OV-3 456 5.430154 0.806
0.848 0.834 0.8094 0.8719 0.891 0.9424 0.903 1.1026
026_945_00288480 Ovary 945 OVMIU 456 5.797932 0.75 0.737 0.748
0.7949 0.7921 0.844 0.8857 0.9368 0.9866 026_948_00291230 Ovary 948
OVTOKO 456 5.89738 0.829 0.912 0.942 0.9105 0.938 0.909 0.9025
0.9329 0.9575 026_8255_00293640 Ovary 8255 OV-56 456 5.989644 0.874
0.894 0.904 0.9159 0.9189 0.911 0.9267 0.9453 0.9277
026_937_00288460 Ovary 937 OAW28 456 6.027686 0.859 0.878 0.802
0.8512 0.7724 0.814 1.0447 0.8649 1.0363 026_1127_00296460 Ovary
1127 NIH:OVCAR-3 456 6.109694 0.886 0.95 0.992 0.9785 0.8788 0.986
0.9619 1.0732 0.8745 026_947_00263500 Ovary 947 OVKATE 456 6.266319
0.819 0.823 0.875 0.8874 0.9093 0.906 0.9175 0.9563 0.9507
026_938_00292910 Ovary 938 OAW42 456 6.299032 0.918 0.963 0.958
0.9614 0.9778 1.041 1.0213 0.9899 1.0087 026_945_00256180 Ovary 945
OVMIU 456 6.360298 0.891 0.987 1.001 0.9225 0.8452 0.776 0.8273
0.9233 1.0102 026_946_00291190 Ovary 946 OVISE 456 6.600815 0.805
0.739 0.737 0.7358 0.7593 0.759 0.8539 0.8664 0.8624
026_8254_00295890 Ovary 8254 OV-17R 456 6.877329 0.913 1.041 0.974
1.0282 1.0077 1.015 1.0376 0.9484 1.0604 026_973_00295540 Pancreas
973 HUP-T4 456 0.866739 0.321 0.315 0.362 0.4721 0.5603 0.814
1.0984 0.8938 1.1407 026_983_00295450 Pancreas 983 SUIT-2 456
1.197889 0.397 0.339 0.382 0.581 0.6565 0.942 0.9937 1.019 1.1926
026_982_00292940 Pancreas 982 QGP-1 456 1.430625 0.538 0.437 0.492
0.5682 0.6307 0.8 0.8372 0.9601 1.0476 026_8118_00295430 Pancreas
8118 MZ1-PC 456 1.48113 0.363 0.489 0.495 0.5335 0.6471 0.671
0.9118 1.008 0.9838 026_8149_00293771 Pancreas 8149 PSN1 456
1.829333 0.531 0.553 0.5 0.5231 0.6104 0.71 0.8375 0.909 0.9509
026_953_00295470 Pancreas 953 AsPC-1 456 1.847893 0.476 0.515 0.508
0.4987 0.5741 0.746 0.8281 1.0377 1.0843 026_1256_00260300 Pancreas
1256 950-MP5 456 1.902786 0.332 0.473 0.515 0.7193 0.8035 0.916
0.9125 1.0441 0.9009 026_976_00298480 Pancreas 976 Panc 04.03 456
2.685969 0.592 0.594 0.57 0.562 0.6645 0.706 0.8319 0.9296 1.0176
026_954_00292870 Pancreas 954 BxPC-3 456 2.90077 0.574 0.607 0.558
0.6572 0.7404 0.813 1.0541 1.0916 0.9092 026_967_00292570 Pancreas
967 Capan-1 456 3.140577 0.522 0.69 0.758 0.8527 0.6713 0.874
0.9506 0.894 0.958 026_975_00295591 Pancreas 975 YAPC 456 3.507979
0.613 0.705 0.797 0.9196 0.8833 0.921 1.0616 1.0323 1.0675
026_1135_00292930 Pancreas 1135 PL18 456 3.513426 0.636 0.623 0.694
0.6978 0.7917 0.832 0.9096 0.9334 0.9721 026_977_00308210 Pancreas
977 KP-1N 456 3.657778 0.607 0.764 0.684 0.7525 0.7655 0.908 0.9944
0.9837 0.9553 026_969_00295580 Pancreas 969 PA-TU-8988T 456
3.657963 0.664 0.653 0.725 0.7145 0.7879 0.992 1.0299 1.0048 1.0863
026_1491_00273490 Pancreas 1491 SNU-324 456 3.732679 0.387 0.683
0.791 0.7879 0.8486 0.832 0.9455 0.9813 0.9695 026_961_00295570
Pancreas 961 Panc 02.03 456 4.088522 0.702 0.699 0.633 0.701 0.7003
0.795 0.8748 0.9019 1.1113 026_974_00292601 Pancreas 974 HUP-T3 456
4.280085 0.684 0.754 0.793 0.909 0.7596 0.88 0.9895 0.9696 1.0099
026_963_00293710 Pancreas 963 Hs 766T 456 4.34359 0.709 0.746 0.833
0.7623 0.8169 1.052 0.9605 0.9551 1.0544
026_959_00292630 Pancreas 959 Panc 03.27 456 4.482408 0.701 0.682
0.805 0.8272 0.7565 0.838 0.9087 0.9434 1.0258 026_981_00293750
Pancreas 981 KP-4 456 4.755774 0.733 0.9 0.784 0.9218 0.8875 1.007
0.9325 1.0003 0.8788 026_968_00292580 Pancreas 968 CFPAC-1 456
4.814885 0.772 0.822 0.87 0.9195 0.9082 0.935 0.8889 1.0106 1.1286
026_1134_00300280 Pancreas 1134 PL4 456 4.852316 0.738 0.739 0.727
0.7443 0.7816 0.797 0.9521 0.9663 1.0376 026_979_00293740 Pancreas
979 KP-3 456 4.900084 0.956 0.777 0.839 0.8825 0.8836 0.909 0.9593
0.9092 0.9686 026_956_00292591 Pancreas 956 HPAF-II 456 5.005523
0.734 0.786 0.762 0.6523 0.7269 0.755 0.9185 0.979 1.0195
026_953_00257150 Pancreas 953 AsPC-1 456 5.141432 0.761 0.708 0.731
0.7295 0.7369 0.79 0.8534 0.8911 0.9577 26_968_00304350 Pancreas
968 CFPAC-1 456 5.151366 0.817 0.828 0.864 0.8724 0.8715 1.008
1.007 0.9779 1.001 026_957_00296350 Pancreas 957 SW 1990 456
5.575766 0.849 0.911 0.906 0.8954 0.9494 0.957 0.977 0.9769 1.1173
026_960_00296311 Pancreas 960 Panc 08.13 456 5.65459 0.875 0.894
0.924 0.9669 0.9243 0.96 1.1005 1.065 1.0309 026_951_00256230
Pancreas 951 HPAC 456 5.736336 0.862 1.024 0.931 0.8532 0.9386
1.035 1.0328 1.0507 1.0612 026_964_00295480 Pancreas 964 Capan-2
456 5.74256 0.877 0.928 0.759 0.8346 0.9905 0.925 1.0364 1.027
0.9499 026_1134_00298560 Pancreas 1134 PL4 456 5.771259 0.844 0.812
0.91 0.8479 0.7738 0.977 0.9068 1.059 0.9774 026_970_00293370
Pancreas 970 PA-TU-8902 456 5.772043 0.841 0.97 0.919 0.8896 0.9177
0.966 0.9181 1.322 0.9871 026_952_00292610 Pancreas 952 MIA PaCa-2
456 5.946631 0.923 0.885 0.718 0.8919 0.8505 1.058 1.0586 0.8995
1.1403 026_972_00296250 Pancreas 972 DAN-G 456 5.955955 0.897 0.924
0.945 0.9646 0.9703 0.978 1.0074 1.032 1.0585 026_951_00295520
Pancreas 951 HPAC 456 5.962773 0.923 0.889 0.787 0.9719 0.9562
1.118 0.986 0.9708 1.1145 026_975_00252910 Pancreas 975 YAPC 456
5.989883 1.015 0.914 0.897 0.8713 0.8903 1.037 0.9777 1.1409 0.981
026_963_00252950 Pancreas 963 Hs 766T 456 6.264463 0.496 0.974
0.953 1.0445 0.9118 1.028 0.9324 1.0245 1.0186 026_958_00296320
Pancreas 958 Panc 10.05 456 6.318194 0.851 0.835 0.851 0.824 0.8929
0.879 0.9003 1.0065 1.0291 026_955_00292640 Pancreas 955 SU.86.86
456 6.593699 1.017 0.82 0.986 0.9378 1.1258 1.072 1.0873 1.0381
0.9789 026_759_00300240 Pleura 759 MSTO-211H 456 3.066599 0.666
0.637 0.664 0.6954 0.8026 0.901 1.0485 1.0165 1.0436
026_1213_00303050 Pleura 1213 H2818 456 3.157415 0.577 0.674 0.728
0.8558 0.9032 0.962 0.9285 1.0744 1.1008 026_8116_00303080 Pleura
8116 MPP-89 456 3.723631 0.416 0.808 0.823 0.9733 0.9887 0.968
1.0012 0.9912 1.0294 026_1206_00302600 Pleura 1206 H2722 456
3.764667 0.604 0.768 0.801 0.8049 0.8692 0.937 0.9073 0.9842 0.9698
026_1210_00302610 Pleura 1210 H2803 456 4.117283 0.686 0.723 0.669
0.7081 0.719 0.892 0.9533 1.0129 0.9547 026_1215_00311260 Pleura
1215 H290 456 4.122283 0.629 0.753 0.69 0.7652 0.7781 0.737 0.8199
0.9853 0.9812 026_1206_00308200 Pleura 1206 H2722 456 4.184817
0.699 0.769 0.85 0.8838 0.889 0.906 1.0135 0.9948 1.0129
026_1212_00300580 Pleura 1212 H2810 456 4.445211 0.69 0.929 0.942
0.9243 0.9927 0.884 0.9289 0.9051 0.9626 026_682_00304600 Pleura
682 NCI-H2452 456 4.452468 0.722 0.847 0.891 0.906 0.9407 0.899
0.9567 1.0786 1.1287 026_1200_00300690 Pleura 1200 H2373 456
4.612295 0.695 0.911 0.887 0.9257 0.9258 0.849 0.9218 0.9772 0.9399
026_1214_00308460 Pleura 1214 H2869 456 4.934927 0.775 0.881 0.878
0.926 0.9241 0.914 0.9475 0.9536 1.0138 026_1214_00308790 Pleura
1214 H2869 456 4.961713 0.802 0.879 0.908 0.9132 0.9528 0.991
0.9959 1.0132 1.0202 026_1202_00303040 Pleura 1202 H2591 456
5.067997 0.82 0.918 0.945 1.0327 0.9338 0.983 0.9541 1.0755 1.1707
026_1209_00300210 Pleura 1209 H28 456 5.341766 0.817 0.821 0.827
0.8331 0.8999 0.927 0.9491 1.0072 0.9857 026_8078_00303060 Pleura
8078 IST-MES1 456 5.358253 0.782 0.897 0.859 0.8822 0.8445 0.907
0.9752 0.926 1.0513 026_1199_00300560 Pleura 1199 H2369 456
5.439636 0.82 0.91 0.99 0.8596 0.9181 0.984 0.9172 0.9896 1.1394
026_8078_00304970 Pleura 8078 IST-MES1 456 5.505825 0.847 0.88
0.853 0.8587 0.8938 0.89 0.9416 0.9423 1.0968 026_1207_00298510
Pleura 1207 H2731 456 5.56988 0.819 1.018 1.003 0.8986 0.9778 0.921
0.9543 0.9856 0.9695 026_1198_00302580 Pleura 1198 H2052 456
5.579483 0.859 0.966 0.995 0.9551 0.9651 1.01 1.0265 1.0233 1.0095
026_1201_00302590 Pleura 1201 H2461 456 5.654117 0.856 0.868 0.914
0.904 0.8852 0.908 0.9343 1.0239 1.1079 026_1213_00300720 Pleura
1213 H2818 456 5.828097 0.843 0.946 0.86 0.9682 0.9236 0.942 0.9493
0.9688 0.9576 026_1211_00300570 Pleura 1211 H2804 456 5.890027
0.881 0.886 0.907 0.8753 0.9153 0.955 0.969 0.9857 1.0448
026_1218_00300730 Pleura 1218 H513 456 5.953268 0.9 0.905 0.905
1.0722 1.0284 0.976 0.9507 1.0341 1.2023 026_1208_00300710 Pleura
1208 H2795 456 6.063798 0.3 0.943 0.961 0.8752 0.9823 0.905 0.9466
0.9643 0.975 026_1203_00300850 Pleura 1203 H2595 456 6.184806 0.92
0.909 0.899 1.0357 1.0632 0.905 0.974 1.018 0.9037
026_8245_00282710 pleural effusion 8245 KMS-11 456 0.702427 0.356
0.381 0.404 0.4804 0.491 0.652 0.7128 1.0645 1.1048
026_996_00298490 Prostate 996 22RV1 456 2.346887 0.542 0.521 0.592
0.6234 0.6635 0.707 0.7764 0.8517 0.9168 026_985_00303070 Prostate
985 LNCaP clone FGC 456 4.018762 0.727 0.863 0.923 1.1164 1.0922
0.984 1.0179 1.1749 1.0533 026_987_00298550 Prostate 987 PC-3 456
4.097696 0.698 0.761 0.871 0.9338 0.9109 0.914 0.9938 1.2109 0.991
026_1001_00300200 Prostate 1001 DU 145 456 5.044916 0.768 0.832
0.867 0.8699 0.8879 0.906 0.9933 0.9168 1.1136 026_988_00308270
Prostate 988 PWR-1E 456 5.228199 0.804 0.854 0.877 0.8566 0.8621
0.923 0.9727 0.9516 0.9806 026_997_00300660 Prostate 997 BPH-1 456
5.448713 0.479 0.868 1.201 1.1209 1.1016 0.674 0.9562 1.1603 0.8983
026_1000_00300740 Prostate 1000 VCaP 456 5.968581 0.692 1.071 1.005
1.0988 0.9203 0.91 0.9225 0.9154 1.1301 026_1009_00264700 Skin 1009
WM35 456 -2.372512 0.112 0.201 0.203 0.2168 0.2138 0.242 0.3676
0.463 0.7025 026_8212_00264590 Skin 8212 CP50-MEL-B 456 -1.651586
0.151 0.398 0.392 0.3817 0.3556 0.35 0.4631 0.5633 0.8209
026_1039_00265170 Skin 1039 SK-MEL-30 456 -1.476493 0.263 0.352
0.317 0.3236 0.3515 0.424 0.5534 0.8565 0.8549 026_1023_00269690
Skin 1023 SK-MEL-2 456 -1.357658 0.147 0.207 0.235 0.2621 0.2935
0.343 0.5053 0.6866 0.8936 026_1034_00264680 Skin 1034 MEL-HO 456
-0.964561 0.144 0.372 0.375 0.3139 0.3537 0.327 0.4189 0.6188
0.8613 026_8073_00263480 Skin 8073 HT-144 456 -0.641233 0.154 0.241
0.236 0.3036 0.3695 0.397 0.6432 0.7444 0.9033 026_8114_00259970
Skin 8114 MMAC-SF 456 -0.613008 0.211 0.399 0.492 0.3813 0.4238
0.662 0.5683 0.6942 1.0563 026_8209_00266540 Skin 8209 A4-Fuk 456
-0.557176 0.179 0.476 0.467 0.4234 0.4176 0.498 0.6644 0.7338
0.7609 026_1046_00263470 Skin 1046 HMVII 456 -0.454742 0.389 0.419
0.403 0.3903 0.4097 0.503 0.6117 0.8285 1.0625 026_8120_00262840
Skin 8120 MZ7-mel 456 -0.254795 0.073 0.323 0.41 0.4001 0.4106
0.468 0.6687 0.6311 0.8761 026_8191_00260580 Skin 8191 UACC-257 456
-0.10266 0.456 0.468 0.426 0.404 0.4006 0.534 0.6654 0.7954 0.894
026_1176_00266430 Skin 1176 451Lu 456 -0.00066 0.26 0.428 0.38
0.386 0.4254 0.499 0.6795 0.8423 0.9729 026_1149_00263460 Skin 1149
G-MEL 456 0.018696 0.065 0.23 0.181 0.1928 0.2465 0.404 0.7707
0.8991 0.9576 026_1147_00263750 Skin 1147 SK-MEL-28 456 0.142676
0.187 0.48 0.376 0.4566 0.4611 0.565 0.6083 0.8116 0.9418
026_1037_00260080 Skin 1037 SK-MEL-1 456 0.168848 0.176 0.417 0.368
0.3716 0.3919 0.613 0.6978 0.9091 0.966 026_1190_00266570 Skin 1190
Hs 939.T 456 0.185332 0.283 0.631 0.615 0.5802 0.5603 0.559 0.5576
0.7187 0.8082 026_1024_00265150 Skin 1024 M-14 456 0.192152 0.418
0.433 0.383 0.391 0.4651 0.618 0.742 0.7678 0.9177
026_1025_00269710 Skin 1025 COLO-679 456 0.228168 0.267 0.288 0.296
0.3664 0.4611 0.681 0.8141 0.9334 1.1196 026_8161_00263740 Skin
8161 SH-4 456 0.433257 0.203 0.383 0.33 0.3147 0.3323 0.488 0.7539
0.8959 0.983 026_8097_00262820 Skin 8097 LB2518-MEL 456 0.437866
0.076 0.501 0.37 0.3843 0.4237 0.706 0.7414 0.8914 0.9396
026_8120_00260550 Skin 8120 MZ7-mel 456 0.447706 0.299 0.428 0.435
0.4782 0.4839 0.498 0.6521 0.7326 0.8788 026_1033_00262470 Skin
1033 IPC-298 456 0.460127 0.363 0.36 0.404 0.4297 0.5245 0.562
0.7868 0.9191 0.8529 026_1031_00264030 Skin 1031 IGR-37 456 0.57267
0.159 0.498 0.512 0.4794 0.5367 0.602 0.8381 0.9754 1.0032
026_1006_00260110 Skin 1006 WM-115 456 0.616077 0.079 0.437 0.583
0.4912 0.4928 0.627 0.6176 0.6485 0.9586 026_8023_00265291 Skin
8023 COLO-829 456 0.740747 0.164 0.659 0.533 0.5534 0.5976 0.709
0.8699 0.9148 0.9877 026_1036_00264690 Skin 1036 RVH-421 456
0.784255 0.099 0.511 0.489 0.5329 0.5785 0.531 0.5954 0.5867 0.7408
026_1011_00269770 Skin 1011 WM278 456 0.944573 0.289 0.463 0.457
0.4545 0.4605 0.568 0.7513 0.8836 0.9597 026_1003_00269650 Skin
1003 G-361 456 0.948854 0.099 0.326 0.434 0.5233 0.6155 0.705 0.83
0.8998 1.0756 026_8002_00263700 Skin 8002 A101D 456 0.960267 0.088
0.468 0.524 0.5105 0.5276 0.55 0.5835 0.7643 0.8791
026_8104_00260540 Skin 8104 LOXIMVI 456 1.072404 0.278 0.403 0.39
0.4769 0.6158 0.839 0.877 0.864 0.9209 026_1005_00268780 Skin 1005
A-375 456 1.191442 0.45 0.499 0.485 0.4604 0.5149 0.506 0.6624
0.892 0.928 026_1004_00260870 Skin 1004 C32 456 1.195832 0.288
0.331 0.256 0.265 0.3055 0.504 0.8497 0.9278 0.9753
026_1030_00264020 Skin 1030 IGR-1 456 1.307792 0.075 0.388 0.556
0.5173 0.6922 0.688 0.8911 0.7819 0.8658 026_1035_00265160 Skin
1035 MEL-JUSO 456 1.414781 0.305 0.481 0.494 0.5153 0.7013 0.739
0.9127 0.9597 1.0093 026_8119_00259980 Skin 8119 MZ2-MEL. 456
1.446447 0.402 0.467 0.438 0.4246 0.4152 0.679 0.7345 0.8985 0.9484
026_8097_00260530 Skin 8097 LB2518-MEL 456 1.447504 0.439 0.438
0.44 0.3578 0.3688 0.635 0.8111 0.8622 0.8731 026_8225_00264640
Skin 8225 SK-MEL-5 456 1.574212 0.07 0.432 0.581 0.5597 0.6534
0.603 0.7648 0.9285 1.0194 026_8104_00262830 Skin 8104 LOXIMVI 456
1.676415 0.172 0.565 0.594 0.5546 0.7085 0.865 1.2098 1.0454 1.1061
026_1042_00274340 Skin 1042 COLO 792 456 1.75025 0.329 0.514 0.573
0.6166 0.6518 0.787 0.8993 0.9066 0.9485 026_1026_00269720 Skin
1026 COLO-783 456 1.791757 0.479 0.599 0.603 0.6144 0.5728 0.634
0.6979 0.8575 0.9077 026_1010_00262530 Skin 1010 WM1552C 456
2.109919 0.364 0.794 0.685 0.6806 0.6737 0.881 0.8586 0.9992 0.9359
026_1041_00263450 Skin 1041 A431 456 2.481335 0.436 0.563 0.633
0.821 0.8517 0.858 1.0792 0.8962 0.9346 026_8098_00269090 Skin 8098
LB373-MEL-D 456 2.623655 0.496 0.615 0.627 0.6225 0.6708 0.815
0.8109 0.9711 1.0201 026_1022_00262940 Skin 1022 RPMI-7951 456
2.647594 0.249 0.552 0.678 0.6664 0.7181 0.824 0.8966 0.8605 0.908
026_1181_00269080 Skin 1181 Hs 944.T 456 2.824283 0.206 0.612 0.779
0.8499 0.8608 0.892 0.9924 1.1546 1.1214 026_1008_00262540 Skin
1008 WM793B 456 2.933974 0.371 0.744 0.693 0.6962 0.7393 0.731
0.8508 0.9586 0.8942 026_1027_00265280 Skin 1027 COLO-800 456
3.132083 0.537 0.717 0.804 0.8959 0.9417 0.926 0.9678 0.954 1.0136
026_8034_00265300 Skin 8034 DJM-1 456 3.234358 0.554 0.778 0.806
0.9452 0.9331 1.005 0.9966 1.0061 1.0084 026_1181_00264620 Skin
1181 Hs 944.T 456 3.491747 0.094 0.698 0.822 0.9152 0.8448 0.9
0.9921 0.9743 0.9841 026_8225_00263510 Skin 8225 SK-MEL-5 456
3.505326 0.234 0.653 0.694 0.7331 0.7681 0.79 0.9216 0.9437 0.9528
026_1145_00263720 Skin 1145 CHL-1 456 3.811805 0.609 0.895 0.833
0.8778 0.9025 0.855 0.9286 0.9316 1.0458 026_8060_00265311 Skin
8060 GAK 456 4.2621 0.37 0.758 0.827 0.7891 0.8733 0.894 0.9899
0.9692 0.9911 026_1047_00265320 Skin 1047 MEWO 456 4.342961 0.779
0.931 0.98 0.9757 0.9449 0.991 1.0008 0.9996 1 026_1022_00260910
Skin 1022 RPMI-7951 456 4.58823 0.257 0.748 0.801 0.8703 0.8751
0.884 0.9495 0.9173 0.9542 026_1038_00260090 Skin 1038 SK-MEL-3 456
4.834702 0.151 0.731 0.777 0.7955 0.7955 0.827 0.8793 0.913 0.9361
026_1002_00260861 Skin 1002 A2058 456 5.928358 0.795 0.897 0.85
0.894 0.8474 0.885 1.0053 0.8902 0.984 026_8025_00306491 Skin 8025
CP66-MEL 456 6.278644 0.914 0.888 1.059 0.9053 1.0334 0.961 0.8798
1.0258 0.9475 026_1120_00264061 Skin 1120 UACC-62 456 6.317961
0.417 0.99 0.941 0.9531 0.9635 1.005 1.0535 1.0033 0.9903
026_1049_00269761 Skin 1049 VMRC-MELG 456 6.363031 0.904 0.991
0.949 0.974 0.9849 1.004 1.032 1.0282 1.0264 026_8077_00311281 Skin
8077 IST-MEL1 456 6.486215 0.817 0.849 0.783 0.8212 0.8361 0.894
0.8282 0.9342 0.9134 026_1002_00262871 Skin 1002 A2058 456 6.768535
0.494 1.023 0.918 0.9957 0.9334 0.899 0.9223 0.931 0.9502
026_1191_00266580 Skin 1191 Hs 940.T 456 7.017835 0.562 0.98 1.123
1.1931 0.9727 1.159 1.1334 1.0446 1.0643 026_8211_00306531 Skin
8211 SK-MEL-24 456 7.240348 0.995 1.062 1.047 0.9379 1.0364 1.075
1.0521 1.0504 1.0137 026_1004_00262880 Skin 1004 C32 456 7.407136
1.327 0.196 1.169 0.1356 1.1547 0.213 1.1935 0.7281 1.1468
026_8077_00263490 Skin 8077 IST-MEL1 456 8.189893 1.25 1.21 1.184
1.1565 0.9453 0.931 1.1642 1.0861 1.0387 026_1076_00260350 Stomach
1076 OCUM-1 456 -1.975727 0.138 0.299 0.203 0.1441 0.1472 0.137
0.1598 0.2644 0.5747 026_1050_00262790 Stomach 1050 AGS 456
0.402284 0.198 0.304 0.337 0.4411 0.6201 0.452 0.6355 0.7729 1.0239
026_1070_00258920 Stomach 1070 HSC-39 456 1.218484 0.163 0.38 0.469
0.5806 0.7048 0.892 1.16 0.922 1.1787 026_8193_00311240 Stomach
8193 ECC10 456 1.328742 0.306 0.42 0.445 0.5445 0.7218 0.758 0.8813
0.9883 0.9993 026_1052_00255810 Stomach 1052 SNU-1 456 1.425654
0.369 0.469 0.497 0.5342 0.5814 0.77 0.7984 0.8285 0.947
026_1056_00316590 Stomach 1056 KATO III 456 1.949149 0.309 0.531
0.622 0.6777 0.7504 0.912 0.9052 1.0407 1.0158 026_1060_00256830
Stomach 1060 MKN45 456 2.285447 0.504 0.586 0.532 0.5224
0.5043 0.577 0.7757 0.8156 0.9028 026_1072_00258880 Stomach 1072
23132/87 456 2.907608 0.473 0.717 0.911 0.8748 1.0846 1.055 1.07
0.981 1.001 026_1060_00262910 Stomach 1060 MKN45 456 2.913103 0.498
0.498 0.474 0.9422 0.5296 0.623 1.0575 0.9958 0.8867
026_1078_00263960 Stomach 1078 IM-95 456 3.209661 0.348 0.625 0.723
1.042 0.7453 0.866 0.8746 0.9835 0.9734 026_1054_00308870 Stomach
1054 SNU-16 456 3.34225 0.574 0.78 0.883 0.9578 0.9599 0.973 0.9861
1.0293 1.027 026_1064_00258960 Stomach 1064 NUGC-3 456 3.516892
0.253 0.754 0.947 0.9636 0.9596 1.019 1.0426 1.0278 1.0027
026_1075_00308260 Stomach 1075 NUGC-4 456 3.652276 0.617 0.827
0.703 0.9984 1.0102 1.068 1.1379 0.9929 1.1512 026_1065_00273570
Stomach 1065 MKN7 456 3.792655 0.621 0.777 0.872 0.891 0.9122 0.904
0.9197 0.9359 0.9547 026_1067_00311560 Stomach 1067 RERF-GC-1B 456
4.166168 0.732 0.677 0.761 0.8438 0.781 0.877 0.8716 0.958 1.0772
26_1067_00314080 Stomach 1067 RERF-GC-1B 456 4.402339 0.708 0.833
0.831 0.8383 0.8843 0.854 0.9212 0.9276 1.1227 026_1068_00258930
Stomach 1068 MKN28 456 4.480011 0.554 0.885 0.897 0.9579 0.9774
1.009 1.0156 1.0042 1.0326 026_1057_00271310 Stomach 1057 Hs 746T
456 4.481195 0.745 0.743 0.677 0.6977 0.6882 0.86 0.875 0.9503
1.0984 026_1060_00260890 Stomach 1060 MKN45 456 4.604315 0.753
0.738 0.731 0.6895 0.6891 0.836 1.0727 1.0008 0.9928
026_1051_00269200 Stomach 1051 FU97 456 4.755599 0.738 0.851 0.914
0.8985 0.8947 0.955 0.9241 0.9866 1.0457 026_8187_00316461 Stomach
8187 TGBC11TKB 456 4.931222 0.775 0.953 0.925 0.9227 0.9488 0.988
0.9649 0.9982 0.9763 026_1058_00263730 Stomach 1058 NCI-N87 456
5.400356 0.754 0.824 0.808 0.8382 0.8486 0.868 0.9105 0.9132 0.9828
026_1064_00264040 Stomach 1064 NUGC-3 456 6.53754 0.915 1.021 1.007
1.0316 0.9966 1.014 1.0431 0.9819 1.0068 026_1077_00264820 Stomach
1077 SCH 456 6.77844 0.466 1.271 1.195 1.2103 1.1255 0.977 0.9179
1.1315 1.1153 026_1051_00265140 Stomach 1051 FU97 456 6.812551
0.149 0.752 0.783 0.8562 0.833 0.758 0.8748 0.8287 0.8438
026_8067_00258320 Stomach 8067 GT3TKB 456 7.007727 0.623 0.977
1.004 1.0575 1.0443 1.026 1.0598 1.0143 1.0267 026_1053_00258360
Stomach 1053 SNU-5 456 7.265878 1 0.967 0.868 0.95 0.958 1.018
1.0763 1.0284 1.0105 026_1073_00258910 Stomach 1073 HGC-27 456
7.474785 0.055 1.003 1.012 1.0288 1.0225 1.013 1.0331 0.9845 0.9972
026_8062_00266170 Stomach 8062 GCIY 456 7.70033 1.076 1.218 0.962
1.0422 0.9155 1.202 0.9466 0.9302 1.1001 026_1059_00256820 Stomach
1059 MKN1 456 7.84127 1.023 1.223 1.097 1.1806 1.0334 0.924 1.0209
0.9691 0.9885 026_8216_00258330 Stomach 8216 RF-48 456 8.041101
0.627 1.056 1.075 1.0873 1.0084 1.031 1.0144 1.008 0.9914
026_8143_00302630 Testes 8143 NTERA-S-cl-D1 456 4.238289 0.709 0.74
0.781 0.8439 0.8942 0.971 0.9553 0.9264 1.0878 026_1081_00299690
Testes 1081 NCC-IT-A3 456 4.69755 0.834 0.925 0.99 1.0022 1.0753
0.959 1.0664 0.9907 0.9674 026_1082_00300430 Testes 1082 NEC8 456
5.794775 0.518 0.919 1.026 1.1263 1.0267 0.882 1.2293 0.8206 0.9481
026_1087_00311210 Thyroid 1087 BHT-101 456 0.094631 0.409 0.442
0.405 0.4025 0.4677 0.606 0.749 0.9156 0.9734 026_1098_00252970
Thyroid 1098 IHH-4 456 0.734299 0.193 0.418 0.365 0.5027 0.4309
0.608 0.6796 0.9903 0.9333 026_1093_00253040 Thyroid 1093
TT2609-C02 456 1.438563 0.33 0.462 0.489 0.4674 0.6027 0.719 0.8623
0.9552 1.0031 026_1100_00252990 Thyroid 1100 KMH-2 456 1.898194
0.142 0.5 0.577 0.6566 0.6074 0.682 0.7694 0.8888 0.9822
026_1085_00252920 Thyroid 1085 8505C 456 2.3348 0.536 0.498 0.605
0.5369 0.584 0.788 0.7985 1.0523 0.937 026_1088_00252930 Thyroid
1088 CAL-62 456 2.495611 0.451 0.785 0.606 0.8114 0.6653 0.836
1.0285 1.0053 0.944 026_1089_00261010 Thyroid 1089 HTC-C3 456
2.538171 0.577 0.55 0.623 0.7757 0.5886 0.616 0.7161 0.7476 0.9625
026_1090_00311330 Thyroid 1090 ML-1 456 2.64388 0.368 0.64 0.793
0.8005 0.8127 0.931 0.9783 0.9389 1.0355 026_1090_00253010 Thyroid
1090 ML-1 456 3.121258 0.165 0.778 0.787 0.6376 0.6432 0.975 1.0524
0.7737 1.0765 026_1090_00308850 Thyroid 1090 ML-1 456 3.18393 0.308
0.673 0.724 0.8112 0.8338 0.925 0.9016 0.8988 1.0407
026_1086_00255730 Thyroid 1086 B-CPAP 456 3.682081 0.171 0.695
0.763 0.7585 0.7239 0.856 0.9452 0.9473 1.0079 026_8020_00314210
Thyroid 8020 CGTH-W-1 456 4.407018 0.694 0.874 0.871 0.8186 0.9478
0.965 0.9497 1.0206 1.0176 026_1084_00266520 Thyroid 1084 8305C 456
4.442179 0.592 0.739 0.746 0.7517 0.73 0.793 0.8273 0.8894 0.8901
026_8213_00259200 Thyroid 8213 TT 456 4.648028 0.769 0.966 0.965
0.9831 0.9968 0.984 0.9765 0.9632 0.942 026_8082_00252980 Thyroid
8082 K5 456 6.244906 0.27 0.935 0.007 0.9319 0.951 0.93 0.997
0.9506 0.9691 026_1099_00306470 Thyroid 1099 ASH-3 456 6.262224
0.912 0.905 0.964 0.9293 0.932 0.965 0.9777 0.9947 1.0629
026_1094_00259110 Thyroid 1094 FTC-133 456 6.573461 0.084 0.958
0.978 1.0951 1.0879 1.057 1.063 1.0254 0.8654 026_1092_00259170
Thyroid 1092 S-117 456 6.767861 0.271 0.973 0.988 1.0669 1.039
1.047 1.018 1.0563 1.0542 026_1097_00259160 Thyroid 1097 RO82-W-1
456 7.698514 0.685 1.079 1.053 1.0554 1.0411 1.049 1.0682 1.0349
1.0563 26_8036_00304360 UrinaryTrack 8036 DSH1 456 1.200178 0.377
0.348 0.397 0.5545 0.7402 0.871 0.9778 0.9884 1.0288
026_24_00298840 UrinaryTrack 24 RT4 456 2.281802 0.504 0.543 0.577
0.6272 0.7473 0.889 1.0564 1.0628 1.0768 026_18_00299720
UrinaryTrack 18 RT-112 456 2.448355 0.479 0.581 0.587 0.6728 0.7768
0.797 0.8705 1.106 1.03 026_24_00252540 UrinaryTrack 24 RT4 456
2.668527 0.216 0.59 0.634 0.6994 0.7144 0.855 0.9368 0.9696 1.0194
026_8036_00257170 UrinaryTrack 8036 DSH1 456 2.728478 0.679 0.605
0.639 0.6941 0.7355 0.829 0.9411 1.0033 1.0113 026_9_00298850
UrinaryTrack 9 SW 780 456 2.896276 0.523 0.587 0.676 0.7538 0.7859
0.866 0.9594 0.9717 1.0348 026_15_00316530 UrinaryTrack 15 BFTC-905
456 3.218144 0.607 0.59 0.688 0.6971 0.8338 0.848 0.8978 1.0448
1.0799 026_8_00298880 UrinaryTrack 8 UM-UC-3 456 3.221517 0.557
0.72 0.789 0.8471 0.8599 0.995 1.0484 1.045 1.0264 026_6_00303090
UrinaryTrack 6 5637 456 3.348554 0.553 0.725 0.744 0.7649 0.9313
0.918 0.9221 0.9563 1.0354 026_6_00256650 UrinaryTrack 6 5637 456
3.837233 0.726 0.703 0.715 0.7187 0.7189 0.755 0.9193 0.965 1.0004
026_8101_00256280 UrinaryTrack 8101 LB831-BLC 456 3.939929 0.504
0.835 0.952 1.0143 1.0014 1.01 0.9848 0.9739 0.9713 026_11_00298870
UrinaryTrack 11 T24 456 4.120815 0.697 0.744 0.763 0.8533 0.8788
0.941 0.9969 1.0222 1.1105 026_19_00298860 UrinaryTrack 19 SW-1710
456 4.713264 0.758 0.847 0.828 0.8254 0.8943 0.979 0.9968 1.0264
1.0955 026_7_00302400 UrinaryTrack 7 SCaBER 456 4.780909 0.779
0.775 0.708 0.7824 0.8163 0.882 1.0463 0.9494 1.0621
026_8101_00302620 UrinaryTrack 8101 LB831-BLC 456 4.856105 0.76
0.882 0.929 0.9235 0.9393 0.926 0.9386 0.9281 1.0507
026_22_00298810 UrinaryTrack 22 HT 1376 456 4.888331 0.823 0.879
1.047 0.8692 1.0283 0.988 1.0731 1.1432 1.097 026_20_00299740
UrinaryTrack 20 VM-CUB1 456 4.987745 0.751 0.753 0.758 0.8989
0.8726 0.877 0.9581 0.9471 0.9762 026_14_00298780 UrinaryTrack 14
647-V 456 5.049696 0.854 0.851 0.873 0.8859 0.9584 1.097 1.1389
1.2074 1.0083 026_12_00299730 UrinaryTrack 12 TCCSUP 456 5.306108
0.821 0.876 0.897 0.9178 0.9315 0.962 0.9176 1.0469 1.0217
026_13_00298770 UrinaryTrack 13 639-V 456 5.658711 0.867 0.933
0.849 0.9317 0.8466 1.046 1.0223 1.1266 1.2765 026_16_00298800
UrinaryTrack 16 CAL-29 456 5.956916 0.811 0.834 0.909 0.8884 0.9073
0.924 0.956 0.9709 0.9841 026_3_00308800 UrinaryTrack 3 HT-1197 456
6.060787 0.877 0.935 0.984 0.8762 0.9541 0.87 1.0393 1.0266 1.0429
026_17_00300420 UrinaryTrack 17 KU-19-19 456 6.388593 0.216 0.953
0.975 0.9996 0.9834 0.744 0.9959 0.9777 1.0473 026_10_00298820
UrinaryTrack 10 J82 456 6.755266 0.951 0.965 0.925 0.9536 1.0393
1.06 1.0882 1.0657 1.037 026_8154_00262510 Uterus 8154 RL95-2 456
0.033885 0.116 0.241 0.307 0.315 0.5516 0.413 0.6863 0.814 0.8778
026_1116_00268860 Uterus 1116 SNG-M 456 2.236003 0.377 0.605 0.635
0.4653 0.7155 0.805 0.9708 1.0702 1.1724 026_8154_00280250 Uterus
8154 RL95-2 456 2.655708 0.475 0.611 0.69 0.5902 0.6865 0.677 0.826
0.8426 0.9723 026_8166_00269750 Uterus 8166 SK-UT-1 456 2.769805
0.399 0.642 0.861 0.9305 0.8691 0.946 0.8217 0.867 0.9308
026_1115_00268850 Uterus 1115 SKN 456 3.232484 0.402 0.724 0.819
0.8345 0.9291 0.941 1.0001 0.9878 0.9814 026_1112_00268820 Uterus
1112 Ishikawa (Heraklio) 02 ER- 456 3.572844 0.62 0.659 0.62 0.6814
0.702 0.72 0.9036 0.8807 1.0744 026_1107_00311310 Uterus 1107
MFE-296 456 3.590101 0.601 0.697 0.647 0.7074 0.7773 0.802 0.8186
0.9166 1.0105 026_1108_00308840 Uterus 1108 MFE-319 456 3.953038
0.7 0.723 0.698 0.8033 0.8993 0.819 0.9773 1.1189 1.0369
026_1109_00269701 Uterus 1109 COLO 684 456 4.059524 0.645 0.818
0.925 0.873 0.8541 0.934 0.9505 0.9782 1.0023 026_1102_00268790
Uterus 1102 AN3CA 456 4.238238 0.395 0.808 0.815 0.8203 0.9291
0.817 0.9786 0.9815 0.9877 026_1108_00314330 Uterus 1108 MFE-319
456 4.375739 0.745 0.784 0.794 0.8526 0.9259 0.907 1.0498 1.0643
1.1339 026_1113_00268840 Uterus 1113 MES-SA 456 4.83984 0.832 0.962
1.003 0.9833 0.9906 0.984 0.9981 1.0121 0.9684 026_1117_00268810
Uterus 1117 HEC-1 456 5.215029 0.824 0.904 0.885 0.8856 0.9834
0.972 0.9217 1.0697 1.0963 026_1113_00318661 Uterus 1113 MES-SA 456
5.307659 0.807 0.995 0.947 0.9632 1.0122 1.033 1.0384 1.0103 0.9636
026_8206_00264270 Uterus 8206 KLE 456 5.888467 0.885 1 1.029 1.0338
1.0454 1.029 0.9964 1.0291 0.9684 026_1105_00308190 Uterus 1105
ESS-1 456 6.082924 0.887 0.915 0.911 0.9349 0.8927 0.94 0.9007
0.9407 0.9198 026_1104_00306500 Uterus 1104 EN 456 7.302741 1.047
0.983 0.999 0.9204 1.0053 1.011 1.0162 1.0749 1.0308
026_1106_00306510 Uterus 1106 MFE-280 456 7.686964 1.124 0.983 1
0.9683 1.0601 1.156 1.0799 1.0206 0.9018 026_8163_00302640 Vulva
8163 SK-LMS-1 456 2.432701 0.479 0.526 0.621 0.7198 0.8786 0.991
1.0278 1.0369 1.1613 026_8173_00302660 Vulva 8173 SW954 456
2.875791 0.539 0.622 0.644 0.6956 0.8294 0.866 1.072 1.0089 1.087
26_8174_00304380 Vulva 8174 SW962 456 4.641057 0.756 0.704 0.824
0.7868 0.8056 0.875 0.851 0.96 1.0607 026_481_00302570 Vulva 481
CAL-39 456 5.164144 0.803 0.856 0.841 0.8864 0.9389 0.924 0.983
0.9909 1.1127 026_8174_00306540 Vulva 8174 SW962 456 6.896884 0.97
0.963 1.002 0.9267 1.0771 0.973 1.0027 1.0435 1.0588
Example 12
BVD-523 Demonstrates In Vivo Antitumor Activity in
BRAF.sup.V600E-Mutant Cancer Cell Line Xenograft Models
[0292] Based on our in vitro findings that BVD-523 reduced
proliferation and induced apoptosis in a concentration-dependent
manner, BVD-523 was administered by oral gavage to demonstrate its
in vivo anti-tumor activity in models with MAPK/ERK-pathway
dependency. Xenograft models of melanoma (cell line A375), and
colorectal cancer (cell line Colo205), were utilized, both of which
harbor a BRAF.sup.V600E mutation.
[0293] In A375 cell line xenografts, BVD-523 efficacy was compared
with the control cytotoxic alkylating agent temozolomide following
14 days of treatment. BVD-523 demonstrated significant
dose-dependent antitumor activity starting at 50 mg/kg twice daily
(BID) (FIG. 31A). Doses of 50 and 100 mg/kg BID significantly
attenuated tumor growth, with tumor growth inhibition (TGI) of 71%
(P=0.004) and 99% (P<0.001), respectively. Seven partial
regressions (PRs) were noted in the 100 mg/kg BID group; no
regression responses were noted in any other group. The efficacy
observed compared favorably with that of temozolomide, which when
administered at 75 and 175 mg/kg resulted in modest dose-dependent
TGI of 34% (P>0.05) and 78% (P=0.005), respectively.
[0294] Additionally, BVD-523 demonstrated antitumor efficacy in a
Colo205 human colorectal cancer cell line xenograft model (FIG.
31B). BVD-523 again showed significant dose-dependent tumor
regressions at doses of 50, 75, and 100 mg/kg BID, yielding mean
tumor regressions T/T.sub.i (T=End of treatment, T.sub.i=Treatment
initiation) of -48.2%, -77.2%, and -92.3%, respectively (all
P<0.0001). Regression was not observed at the lowest dose of
BVD-523 (25 mg/kg BID); however, significant tumor growth
inhibition, with a T/C (T=Treatment, C=Control) of 25.2%
(P<0.0001), was observed. Although not well tolerated, the
positive control chemotherapeutic agent irinotecan (CPT-11) showed
significant antitumor activity, inhibiting Colo205 tumor growth
with a T/C of 6.4% (P<0.0001). However, even at its maximum
tolerated dose in mice, CPT-11 was not as effective as BVD-523 at
doses of 50, 75, or 100 mg/kg BID.
[0295] To establish the relationship between pharmacokinetics and
pharmacodynamics, BVD-523 plasma concentrations were compared with
pERK1/2 levels measured in the tumor by immunohistochemistry and
isotope-tagged internal standard mass spectrometry over a 24-hour
period following a single 100 mg/kg oral dose of BVD-523 (FIG.
31C). Phosphorylation of ERK1/2 was low in untreated tumors (0
hours). Following treatment with BVD-523, ERK1/2 phosphorylation
steadily increased from 1 hour post-dose to maximal levels at 8
hours post-dose, then returned to pre-dose levels by 24 hours. This
increase in pERK1/2 correlated with BVD-523 drug plasma
concentrations. The in vivo observation of increased pERK1/2 with
BVD-523 treatment is consistent with earlier in vitro findings
(FIG. 30D).
Example 13
BVD-523 Results in ERK1/2 Substrate Inhibition Despite Increased
ERK1/2 Phosphorylation
[0296] To examine the effects of BVD-523 on signaling relative to
other known ERK1/2 inhibitors (SCH772984, GDC-0994, and Vx-11e)
(Morris et al. 2013 and Liu et al. 2015), a large-scale reverse
phase protein array (RPPA) of approximately 40 proteins was
employed in a variety of cell lines with sensitivity to ERK
inhibition. Cell lines with common alterations in BRAF and RAS were
assayed: BRAF.sup.V600E mutant lines A375, Colo205, and HT29;
KRAS.sup.G12C-mutant cell line MIAPACa-2; KRAS.sup.G13D-mutant cell
line HCT116; and AN3Ca with atypical HRAS.sup.F82L mutation.
Changes in protein levels are shown as a percentage change from
dimethyl sulfoxide (DMSO)-treated parental control (FIG. 32A and
Table 23). All ERK inhibitors elicited qualitatively similar
protein effects, with the exception of phosphorylation of ERK1/2
(pERK1/2 [ERK1/2-T202, -Y204]); SCH7722984 inhibited pERK1/2 in all
cell lines, while BVD-523, GDC-0994, and Vx-11e markedly increased
pERK1/2. Phospho-p90 RSK (pRSK1) and cyclin D1, which are proximal
and distal targets of pERK1/2, respectively, were similarly
inhibited by all inhibitors tested regardless of the degree of
ERK1/2 phosphorylation (FIG. 32B). These independent findings for
BVD-523 are consistent with studies showing that phosphorylation of
ERK1/2 substrates RSK1/2 remained inhibited despite dramatically
elevated pERK1/2 by Western blots in A375 cells (FIG. 32D), in
addition to protein-binding studies demonstrating BVD-523 binding
and stabilization of pERK1/2 and inactive ERK1/2 (FIG. 29E and FIG.
29F). Therefore, measuring increased pERK1/2 levels could be
considered as a clinical pharmacodynamic biomarker for BVD-523,
while quantifying inhibition of ERK1/2 targets such as pRSK1 and
DUSP6 as well could serve a similar purpose.
[0297] Additional protein changes are of note in this RPPA dataset
(FIG. 32A). Decreased pS6-ribosomal protein appears to be another
pharmacodynamic marker of ERK1/2 inhibition, as evidenced in all
cell lines with all compounds (FIG. 32B). Furthermore, prominent
induction of pAKT appears to be a cell line-dependent observation,
where each ERK1/2 inhibitor induced pAKT in cell lines A375 and
AN3CA cells (FIG. 33). Interestingly, the degree of inhibition of
survival marker pBAD appears to differ between compounds, with only
modest inhibition of pBAD by GDC-0994 compared with the other
ERK1/2 inhibitors tested (FIG. 32A).
[0298] Next, how BVD-523 affects cellular localization of ERK1/2
and downstream target pRSK in a BRAF.sup.V600E-mutant RKO
colorectal cell line (FIG. 32C) was investigated. In resting cells,
ERK1/2 localizes to the cytoplasm, and once stimulated pERK1/2
migrates to target organelles, particularly the nucleus where
transcriptional targets are activated (Wainstein et al. 2016). In
DMSO-treated control cells, pERK1/2 is evident in both nuclear and
cytoplasmic fractions, which is likely reflective of MAPK pathway
activity due to the presence of BRAF.sup.V600E in this cell line.
Treatment with BVD-523 resulted in elevated pERK1/2 in the nucleus
and cytoplasm as well as a modest increase in nuclear total ERK1/2
compared with DMSO-treated cells, suggesting that compound-induced
stabilization of pERK1/2 stimulates some nuclear translocation.
Despite increased pERK1/2 in both compartments, pRSK levels are
lower in the cytoplasmic and nuclear compartments compared with
DMSO control. Comparator MAPK signaling inhibitors (i.e.,
trametinib, SCH7722984, dabrafenib) inhibited phosphorylation of
ERK1/2 and RSK, as reflected by lower levels in the nuclear and
cytoplasmic compartments. These data again suggest that
BVD-523-associated increases in pERK1/2 are evident in both the
cytoplasm and nucleus; however, this does not translate to
activation of target substrates. This is consistent with data
presented in FIG. 30D and FIG. 32A.
TABLE-US-00023 TABLE 23 % change from DMSO (matched cell line)
Treat- Avg(S6 Ribo Avg(S6 Ribo Avg(Cyclin Avg(p90 Avg(bad Avg(4ebp1
Cell Line ment Prot S235 236) Prot S240 244) D1) RSK S380) S112)
T70) A375 BVD -95.3 -91.98 -81.45 -71 -72.37 -31.82 A375 Vx (Empty)
(Empty) -85.46 -65.25 -69.29 23.33 A375 GDC -87.61 -80.3 -81.65
-60.74 -55.32 -29.47 A375 SCH -94.71 -91.78 -84.05 -72.44 -71.44
-31.75 AN3Ca BVD -43.41 -22.2 -0.69 -28.11 -54.1 32.17 AN3Ca Vx
-18.54 -12.55 -9.59 -28.41 -45.63 16.67 AN3Ca GDC -30.74 -23.47
0.34 -29.44 2.53 11.28 AN3Ca SCH -61.99 -35.88 -11.33 -40.26 -39.14
61.57 COLO205 BVD -96.15 -97.33 -23.65 -50.84 -31.51 -36.35 COLO205
Vx -93.04 -94.89 -39.79 -58.6 -30.28 -43.71 COLO205 GDC -91.19
-91.59 -28.02 -57.5 -6.12 -36.69 COLO205 SCH -94.67 -95.09 -36.7
-62.31 -29.4 -27.51 HCT116 BVD -96.31 -96.26 -69.62 -31.81 -34.27
4.03 HCT116 Vx -90.03 -86.06 -72.72 -33.05 -23.88 10.35 HCT116 GDC
-94.82 -95.1 -63.59 -22.25 -12.36 20.5 HCT116 SCH -93.86 -91.07
-73.21 -33.7 -31.29 5.6 HT29 BVD -44.68 -25.67 -37.21 -60.5 -20.66
-41.47 HT29 Vx -32.8 -24.35 -35.2 -43.41 -35.62 -2.89 HT29 GDC
-41.45 -21.74 -35.69 -30.59 -12.98 1.95 HT29 SCH -44.9 -25.73
-36.66 -53.88 -33.9 -40.58 MIAPaca2 BVD -79.46 -88.03 -37.9 -35
-30.29 -9.42 MIAPaca2 Vx -63.36 -74.82 -33.96 -39.91 -20.85 -15.72
MIAPaca2 GDC -67.9 -75.59 -31.92 -39.09 -10.08 -34.01 MIAPaca2 SCH
-77.57 -86.61 -39.88 -38.58 -33.07 19.27 % change from DMSO
(matched cell line) Treat- Avg(p70 S6 Avg(p70 S6 Avg(ERK
Avg(Caspase Cell Line ment Kinase T389 T412) Kinase S371 S394) 1 2)
Avg(Akt) Avg(Raptor) 3 CL D175) A375 BVD -23.79 -8.58 -22.91 -14.03
-7.77 -8.06 A375 Vx -25.54 -17.32 -5.39 -30.34 -12.7 -9.27 A375 GDC
-31.9 -17.34 31.55 -20.7 -14.32 -16.74 A375 SCH -42.73 -28.72
-21.65 -23.26 -11.66 -9.87 AN3Ca BVD -14.78 32.26 -9.05 -22.43
-13.82 -10.8 AN3Ca Vx 0.56 44.04 -11.27 -24.62 -2.47 -12.7 AN3Ca
GDC 26.01 29.09 -2.87 -26.04 -8.05 1.55 AN3Ca SCH -16.63 24.56
-9.27 -16.35 -11.09 1.25 COLO205 BVD -36.4 -18.11 -18.83 -3.85
-7.14 -3.18 COLO205 Vx -28 -13.64 -12.32 -12.51 -0.05 -2.67 COLO205
GDC -32.2 -13.02 -3.33 -11.83 -5.48 23.06 COLO205 SCH -30.4 -14.59
-31.87 -10.31 -2.2 14.08 HCT116 BVD -28.11 -16.9 -29.42 4.41 -7.06
-10.11 HCT116 Vx -20.99 -9.89 -24.01 -18.15 -4.32 -5.19 HCT116 GDC
-24.73 -11.47 -1.9 -6.13 -6.2 -8.36 HCT116 SCH -24.63 -12.3 -10.22
-9.86 -9.66 -4.63 HT29 BVD -24.58 -35.94 -44.3 -13.41 -8.53 -7.03
HT29 Vx -12.31 -22.86 0.24 -17.84 -6.53 -2.86 HT29 GDC -20.86
-25.73 4.66 -10.01 -6.85 -3.44 HT29 SCH -9.55 -20.52 -37 -16.93
-12.18 -7.9 MIAPaca2 BVD -39.23 -28.27 -40.33 23.63 21.15 22.35
MIAPaca2 Vx -30.66 -30.35 -14.85 -0.15 5.4 6.17 MIAPaca2 GDC -40.99
-14.4 -6.88 4.33 22.43 10.47 MIAPaca2 SCH -50.97 -40.47 -23.09
13.47 17.66 21.05 % change from DMSO (matched cell line) Treat-
Avg(mTOR Avg(Bad Avg(c Avg(p70 Avg(Raptor Avg(Bcl Cell Line ment
S2448) S155) Fos) Avg(Rictor) S6 Kinase) S792) 2 T56) A375 BVD
-27.87 -21.9 4.39 -20.11 -7.6 -10.33 -8.54 A375 Vx -21.66 -6.3
-13.65 -8.16 -6.42 -0.86 -7.53 A375 GDC -23.61 -13.31 -12.46 -23.29
-18.11 -15.93 -4.1 A375 SCH -26.17 -13.86 -12.51 -22.13 -17.66
-6.89 -19.55 AN3Ca BVD -10.79 0.66 -5.15 -4.52 -10.27 -8.47 -12.85
AN3Ca Vx -2.37 4.59 -5.52 0.02 -2.33 0.37 -11.73 AN3Ca GDC -2.96
17.31 -0.63 -9.21 -3.67 4.85 1.23 AN3Ca SCH -4.84 12.92 -9.18
-10.89 -7.71 -4.03 -10.73 COLO205 BVD -23.51 -18.18 -12.25 -5.21
0.14 0.41 -11.84 COLO205 Vx -8.52 -9.72 -19.34 1.65 -3.42 0.2
-12.02 COLO205 GDC -7.36 -9.11 -21.33 -5.04 5.83 -9.04 -4.6 COLO205
SCH -9.44 -10.96 -15.17 -19.07 -2.85 -4.17 -6.73 HCT116 BVD -12.78
-30.72 -14.08 -13.05 -12.86 -22.04 -8.36 HCT116 Vx -10.12 -15.59
-13.89 1.78 -4.45 -11.21 -25 HCT116 GDC -19.33 -19.71 -10.36 -10.98
-9.9 -15.77 -11.96 HCT116 SCH -16.05 -22.96 -15.27 -18.5 -14.12
-18.18 -15.89 HT29 BVD -20.68 -25.9 -13.48 -18.76 -10.64 -10.48
-8.18 HT29 Vx -13.94 -11.26 -8.23 -2.6 -1.72 -2.13 -25.01 HT29 GDC
-11.44 -6.7 -12.92 -10.62 -2.83 -1.75 1.61 HT29 SCH -22.65 -7.98
-9.26 -13.03 -2.36 -6.38 -17.9 MIAPaca2 BVD -11.73 -5.65 -18.44
-11.11 -3.59 5.95 -1.78 MIAPaca2 Vx -5.38 7.38 -4.43 -13.24 -9.27
-3.74 -9.86 MIAPaca2 GDC 11.84 8.18 -16.12 -11.25 -3.25 2.18 -2.12
MIAPaca2 SCH -11.48 -1.71 -8.99 -15.15 -7.51 4.43 -1.98 % change
from DMSO (matched cell line) Treat- Avg(Bcl2 Avg(Bcl Avg(mTOR
Avg(Tuberin Avg(Chk1 Avg(CREB Cell Line ment S70) 2) Avg(Bax)
S2481) TSC2 Y1571) S345) S133) A375 BVD -7.91 -12.55 -6.98 -3.36
-6.32 -7.43 -12.65 A375 Vx -4.02 -9.93 -5.05 -2.9 -3.55 0.56 -15.8
A375 GDC 6.3 -3.13 3.97 -19.37 -16.74 -18.35 -1.18 A375 SCH -12.91
-18.53 -17.32 -15.82 -13.7 -10.59 -1.55 AN3Ca BVD -14.81 -16.83
-10.47 2.75 2.86 -13.34 1.03 AN3Ca Vx -14.39 -18.77 -7.89 7.22 3.8
-9.54 0.16 AN3Ca GDC -0.65 7.8 23.83 25.42 25.18 15.46 5.03 AN3Ca
SCH -12.42 -5.14 0.15 11.24 13.44 14.56 3.64 COLO205 BVD -10.2
-0.13 0.28 -1.33 -3.48 -4.63 -2.67 COLO205 Vx -20.82 -14.09 -9.16
-9.43 -9.45 13.12 -10.64 COLO205 GDC -9 17.95 8.2 0.96 0.66 6.08
-11.85 COLO205 SCH -18.52 -14.56 -6.44 -6.68 -1.38 1 -14.56 HCT116
BVD -8.23 -3.59 -0.8 -34.09 -29.89 -39.65 -30.64 HCT116 Vx -17.79
-17.3 -20.46 -17.82 -16.51 -16.49 -9.17 HCT116 GDC -12.53 -4.56
-20.11 -18.34 -9.4 -18.77 -8.56 HCT116 SCH -18.34 -10.58 -13.86
-22.53 -16.17 -18.51 -24.18 HT29 BVD 2.83 10.68 8.39 -8.64 0.66
-13.48 -3.46 HT29 Vx -12.55 -16.73 -18.47 0.52 2.85 6.44 14.16 HT29
GDC 10.59 18.16 1.1 3.23 11.86 6.88 9.54 HT29 SCH -11.45 -10.01
-20.59 13.93 13.25 19.05 9.87 MIAPaca2 BVD 0.89 5.86 23.44 -0.24
1.7 1.2 -19.26 MIAPaca2 Vx -7.83 -2.3 4.33 -3.12 -0.01 2.09 -13.28
MIAPaca2 GDC -6.78 -7.8 3.48 3.65 5.71 32.96 -21.37 MIAPaca2 SCH
-4.23 4.96 15.67 2.43 8.86 13.39 -6.63 % change from DMSO (matched
cell line) Treat- Avg(Caspase Avg(MSK1 Avg(c Cell LIne ment 7 Cl
D198) Avg(Stat3) Avg(Bak) S360) Avg(mTOR) Myc) Avg(Stat1) Avg(Mcl1)
A375 BVD 3.18 6.02 -3.26 3.35 0.2 2.56 1.32 10.67 A375 Vx -7.62
1.71 0.65 -7.63 -3.31 -0.97 0.23 1.6 A375 GDC -5.53 14.24 2.76
-5.21 -6.2 -9.11 -5.67 -9.09 A375 SCH -6 -1.42 -2.37 0.97 -5.18
1.86 -6.01 2.11 AN3Ca BVD -12.35 -4.97 3.38 0.49 5.58 -0.58 -5.65
-4.9 AN3Ca Vx -8.09 -7.67 -1.6 6.83 -6.27 -5.47 -0.83 -4.24 AN3Ca
GDC -15.56 -13.05 7.33 6.84 -2.85 3.99 9.15 -4.16 AN3Ca SCH -24.17
-6.01 9.73 -5.02 -5.33 0.88 5.52 -7.15 COLO205 BVD 6.76 -2.58 3.79
14.69 12.77 -1.11 0.53 -2.7 COLO205 Vx 0.03 -7.96 -2.79 6.22 3.62
-7.42 -6.1 -10.64 COLO205 GDC 1.37 -0.86 6.58 0.82 3.39 -6.22 7.26
2.5 COLO205 SCH 12.36 -6.1 4.59 7.54 -5.62 -0.64 -4.31 -0.14 HCT116
BVD -21.09 -5.52 -13.16 -13.45 -11.96 -11.15 -13.76 -8.72 HCT116 Vx
-11.41 -7.76 -3.07 -13.73 -0.59 -18.56 -14.06 -7.79 HCT116 GDC
-13.99 -0.25 -1.74 -12.93 -4.85 -12.03 -6.32 -4.77 HCT116 SCH
-16.35 -7.67 -4.66 -15.32 -12.56 -12.86 -10.74 -6.55 HT29 BVD -6.93
-6.46 -1.42 1.49 5.02 17.85 4.74 -2.83 HT29 Vx 8.28 -4.28 2.69 5.64
16.94 10.71 4.29 1.36 HT29 GDC -0.88 -3.69 3.56 -0.04 10.45 7.56
2.58 3.71 HT29 SCH 7.02 -2.15 2.78 1.25 9.31 13.55 5.79 14.05
MIAPaca2 BVD -6.44 -1.23 -15.15 -10.72 -3.27 -8.77 8.62 -7.27
MIAPaca2 Vx -0.32 0.61 -1.93 -0.87 -1.58 -5.94 -1.64 -6.58 MIAPaca2
GDC -4.92 -9.44 -8.11 -5.25 -9.8 -7.84 7.4 -1.79 MIAPaca2 SCH 8.35
-9.19 -10.88 2.74 1.64 2.11 7.17 -1.88 % change from DMSO (matched
cell line) Treat- Avg(Bad Avg(Akt Avg(ERK Cell Line ment S136)
Avg(Chk1) Avg(Bim) S473) 1 2 T202 Y204) A375 BVD 12.31 16.94 14.57
73.07 43.34 A375 Vx 5.06 -0.86 12.32 93.85 128.93 A375 GDC -6.21
-7.72 9.8 53.66 142.37 A375 SCH 5.32 2.83 17.52 58.13 -90.63 AN3Ca
BVD -1.02 -8.45 -12.57 52.11 733.27 AN3Ca Vx -3.81 -0.46 -1.46
56.17 718.94 AN3Ca GDC 0.71 9.43 -11.63 82.37 645.51 AN3Ca SCH 5.26
1.35 -14.09 66.17 19.75 COLO205 BVD 5.05 -2.86 41.73 -5.78 14.39
COLO205 Vx -1.01 -9.16 34.1 -10.96 98.48 COLO205 GDC 4 5.06 20.59
4.45 20.01 COLO205 SCH 1.88 7.49 29.22 -1.74 -91.43 HCT116 BVD
-11.49 -11.26 12.44 -6.14 849.12 HCT116 Vx -5.39 -8.63 4.82 -16.94
873.33 HCT116 GDC -1.24 2.28 6.2 4.69 526.64 HCT116 SCH -10.55
-5.58 1.95 -8.76 -75.21 HT29 BVD 10.73 7.4 2.81 -3.06 54.82 HT29 Vx
5.53 4.68 3.12 -20.5 435.68 HT29 GDC 2 5.68 7.74 -10.5 268.99 HT29
SCH 9.67 14.64 0.69 -22.14 -74.84 MIAPaca2 BVD -4.12 -3.51 -12.51
9.9 209.14 MIAPaca2 Vx 0.36 -1.8 2.1 0.48 729.27 MIAPaca2 GDC 2.25
6.56 5.07 2.24 199.59 MIAPaca2 SCH 8.62 12.07 2.08 2.84 -76.71
Example 14
BVD-523 Exhibited Activity in In Vitro Models of BRAF and MEK
Inhibitor Resistance
[0299] Emergence of resistance to BRAF and MEK inhibitors limits
their clinical efficacy. Here, the experiments sought to model and
compare the development of resistance to BRAF (dabrafenib), MEK
(trametinib), and ERK1/2 (BVD-523) inhibition in vitro. Over
several months, BRAF.sup.V600E-mutant A375 cells were cultured in
progressively increasing concentrations of each inhibitor.
Drug-resistant A375 cell lines were readily obtained following
growth in high concentrations of trametinib or dabrafenib, while
developing cell lines with resistance to BVD-523 proved challenging
(FIG. 34A). Overall, these in vitro data suggest that at
concentrations yielding similar target inhibition, resistance to
BVD-523 is delayed compared with dabrafenib or trametinib, and may
translate to durable responses in the clinic.
[0300] Reactivation and dependence on ERK1/2 signaling is a common
feature of acquired resistance to BRAF/MEK inhibition (Morris et
al. 2013 and Hatzivassiliou et al. 2012); therefore, the activity
of BVD-523 in in vitro models of acquired resistance was evaluated.
First, a dabrafenib and trametinib combination-resistant A375
population was obtained using the increased concentration method
described. The IC.sub.50 and IC.sub.50-fold change from parental
A375 for dabrafenib, trametinib, and BVD-523 in the BRAF/MEK
combination-resistant population is shown in Table 24. BVD-523
IC.sub.50 was modestly shifted (2.5-fold), while dabrafenib and
trametinib were more significantly shifted (8.5-fold and 13.5-fold,
respectively) (Table 24). The cytotoxic agent paclitaxel was tested
as a control with only a modest shift in potency observed. These
data support the investigation of BVD-523 in the setting of
BRAF/MEK therapy resistance, although the mechanism of resistance
in this cell population remains to be characterized.
TABLE-US-00024 TABLE 24 BVD-523 activity in models of BRAF/MEK
inhibition Cell Line Dabrafenib Trametinib BVD-523 Paclitaxel
Parental 2.1 0.2 129 1.9 (IC.sub.50 nM) BRAFi- + 17.9 2.7 323 3.5
MEKi-resistant (IC.sub.50 nM) Fold +8.5 +13.5 +2.5 +1.8 Change
[0301] To further investigate the tractability of ERK1/2 inhibition
in a model with a known mechanism of BRAF inhibitor resistance,
AAV-mediated gene targeting was used to generate a pair of RKO
BRAF.sup.V600E-mutant cell lines isogenic for the presence or
absence of an engineered heterozygous knock-in of
MEK1.sup.Q56P-activating mutation (Trunzer et al. 2013 and Emery et
al. 2009). MEK1/2 mutations, including MEK1.sup.Q56P, have been
implicated in both single-agent BRAF and combination BRAF/MEK
therapy-acquired resistance in patients (Wagle et al. 2011, Wagle
et al. 2014, Emery et al. 2009 and Johnson et al. 2015).
Single-agent assays demonstrated that relative to the parental
BRAF.sup.V600E::MEK1 wt cells, the double-mutant
BRAF.sup.V600E::MEK1.sup.Q56P cells displayed a markedly reduced
sensitivity to the BRAF inhibitors vemurafenib and dabrafenib and
the MEK inhibitor trametinib (FIG. 34B). In contrast, response to
BVD-523 was essentially identical in both the parental and
MEK.sup.Q56P-mutant cells, indicating that BVD-523 is not
susceptible to this mechanism of acquired resistance. These results
were confirmed in 2 independently derived double-mutant
BRAF.sup.V600E::MEK1.sup.Q56P cell line clones, thus validating
that results were specifically related to the presence of the
MEK1.sup.Q56P mutation rather than an unrelated clonal artifact
(data not shown). Similar results were also observed with a second
mechanistically distinct ERK1/2 inhibitor (SCH772984), supporting
the expectation that these observations are specifically related to
mechanistic inhibition of ERK1/2 and not due to an off-target
compound effect.
[0302] To further characterize the mechanistic effects of BVD-523
on MAPK pathway signaling in BRAF.sup.V600E::MEK1.sup.Q56P cell
lines, protein levels were assessed by Western blot (FIG. 34C). In
the parental BRAF.sup.V600E RKO cells, a reduced level of pRSK1/2
was observed following 4-hour treatment with BRAF (vemurafenib),
MEK (trametinib), or ERK1/2 (BVD-523) inhibitors at
pharmacologically active concentrations. In contrast, isogenic
double-mutant BRAF.sup.V600E::MEK1.sup.Q56P cells did not exhibit
reduced RSK phosphorylation following BRAF or MEK inhibitor
treatment, while BVD-523 remained effective in inhibiting pRSK1/2
to a level comparable to parental RKO. Similarly, pRB is reduced,
indicating G0/G1 arrest, by 24 hours of BVD-523 treatment in both
parental RKO and BRAF.sup.V600E::MEK1.sup.Q56P.
[0303] Acquired KRAS mutations are also known drivers of resistance
to MAPK pathway inhibitors. To understand the susceptibility of
BVD-523 to this mechanism of resistance, an isogenic panel of
clinically relevant KRAS mutations in colorectal cell line SW48 was
used. Sensitivity to BVD-523 was compared with MEK inhibitors
selumetinib and trametinib (FIG. 34D). Sensitivity to paclitaxel
was unaltered (FIG. 37A). While several mutant KRAS alleles
conferred robust to intermediate levels of resistance to MEK
inhibition, sensitivity to BVD-523 was unaltered by the majority of
alleles, and where a shift in sensitivity was observed, it was not
to the extent observed with trametinib or selumetinib. Overall,
these data suggest that BVD-523 is more efficacious in this context
than MEK inhibitors.
Example 15
BVD-523 Demonstrates In Vivo Activity in a BRAF Inhibitor-Resistant
Patient-Derived Melanoma Xenograft Model
[0304] To confirm and extend the antitumor effects of BVD-523
observed in in vitro models of BRAF-/MEK-acquired resistance, a
BRAF-resistant xenograft model derived from a patient with
resistance to vemurafenib was utilized. BVD-523 was dosed by oral
gavage at 100 mg/kg BID for 28 days, both alone and in combination
with dabrafenib at 50 mg/kg BID (FIG. 35). As expected, minimal
antitumor activity was demonstrated for single-agent dabrafenib
(22% TGI). BVD-523 activity was significant compared with vehicle
control (P.ltoreq.0.05), with a TGI of 78%. In this model,
combining BVD-523 with dabrafenib resulted in a TGI of 76%
(P.ltoreq.0.05); therefore, further benefit was not gained for the
combination compared with single-agent BVD-523 in this model of
BRAF-acquired resistance.
Example 16
Combination Therapy with BVD-523 and a BRAF Inhibitor Provides
Promising Antitumor Activity
[0305] Patients with BRAF-mutant cancer may acquire resistance to
combined BRAF/MEK therapy (Wagle et al. 2014), warranting
consideration of other combination approaches within the MAPK
pathway. The anti-proliferative effects of combining BVD-523 with
the BRAF inhibitor vemurafenib was assessed in the
BRAF.sup.V600E-mutant melanoma cell line G-361. As anticipated,
single agents BVD-523 and vemurafenib were both active, and modest
synergy was observed when combined (FIG. 37B). This indicates that
BVD-523 combined with BRAF inhibitors are at least additive and
potentially synergistic in melanoma cell lines carrying a
BRAF.sup.V600E mutation. Furthermore, generating acquired
resistance in vitro following continuous culturing of
BRAF.sup.V600E mutant cell line (A375) in BRAF inhibitor plus
BVD-523 was challenging. In contrast generating resistance to
dabrafenib alone occurred relatively rapidly (FIG. 37C). Even
resistance to combined dabrafenib and trametinib emerged before
dabrafenib plus trametinib.
[0306] The benefit of combined BRAF and ERK inhibition may not be
fully realized in in vitro combination studies where concentrations
are not limited by tolerability. To understand the benefit of the
combination, efficacy was assessed in vivo utilizing xenografts of
the BRAF.sup.V600E-mutant human melanoma cell line A375. Due to the
noteworthy response to combination treatment, dosing in the
combination groups was stopped on Day 20 to monitor for tumor
regrowth, and was reinitiated on Day 42 (FIG. 36A). Tumors were
measured twice weekly until the study was terminated on Day 45. The
median time to endpoint (TTE) for controls was 9.2 days, and the
maximum possible tumor growth delay (TGD) of 35.8 days was defined
as 100%. Temozolomide treatment resulted in a TGD of 1.3 days (4%)
and no regressions. The 50- and 100-mg/kg dabrafenib monotherapies
produced TGDs of 6.9 days (19%) and 19.3 days (54%), respectively,
a significant survival benefit (P<0.001), and 1 PR in the
100-mg/kg group. The 100-mg/kg BVD-523 monotherapy resulted in a
TGD of 9.3 days (26%), a significant survival benefit (P<0.001),
and 2 durable complete responses. The combinations of dabrafenib
with BVD-523 each produced the maximum possible 100% TGD with
noteworthy regression responses, and statistically superior overall
survival compared with their corresponding monotherapies
(P<0.001). The lowest dose combination produced a noteworthy
7/15 tumor-free survivors (TFS), and the 3 higher-dosage
combinations produced a total of 43/44 TFS, consistent with
curative or near-curative activity (FIG. 36B). In summary, the
combination of dabrafenib with BVD-523 produced a greater number of
TFS and superior efficacy to either single agent.
[0307] Based on the activity of BVD-523 plus dabrafenib in A375
xenograft models with a starting tumor volume of approximately
75-144 mm.sup.3, a follow-up experiment was conducted to determine
the efficacy of combination therapy in "upstaged" A375 xenografts
(average tumor start volume, 700-800 mm.sup.3) (FIG. 36C). The
median TTE for controls was 6.2 days, establishing a maximum
possible TGD of 53.8 days, which was defined as 100% TGD for the
60-day study. BVD-523 100-mg/kg monotherapy produced a negligible
TGD (0.7 day, 1%) and no significant survival difference from
controls (P>0.05). The distribution of TTEs and 2 PRs suggested
there may have been a subset of responders to treatment with
BVD-523 alone. Dabrafenib 50-mg/kg monotherapy was efficacious,
yielding a TGD of 46.2 days (86%) and a significant survival
benefit compared with controls (P<0.001). This group had 5 PRs
and 5 CRs, including 3 TFS, among the 11 evaluable mice (FIG. 36D).
Both combinations of dabrafenib with BVD-523 produced the maximum
100% TGD and a significant survival benefit compared with controls
(P<0.001). Each combination produced 100% regression responses
among evaluable mice, though there were distinctions in regression
activity. The 25-mg/kg dabrafenib and 50-mg/kg BVD-523 combination
had 2 PRs and 8 CRs, with 6/10 TFS, whereas the 50-mg/kg dabrafenib
and 100-mg/kg BVD-523 combination had 11/11 TFS on Day 60 (FIG.
36D). Overall, these data support the rationale for frontline
combination of BVD-523 with BRAF-targeted therapy in
BRAF.sup.V600-mutant melanoma, and this is likely to extend to
other tumor types harboring this alteration.
Discussion
[0308] BVD-523 is a potent, highly selective, reversible, small
molecule ATP-competitive inhibitor of ERK1/2 with activity in in
vivo and in vitro cancer models. In vitro, BVD-523 demonstrated
potent inhibition against several human tumor cell lines,
particularly those harboring activating mutations in the MAPK
signaling pathway, consistent with its mechanism of action. BVD-523
elicited changes in downstream target and effector proteins,
including inhibition of direct substrate of ERK1/2, pRSK, and total
DUSP6 protein levels. These findings are in line with those of
previous studies of other ERK1/2 inhibitors, which demonstrated
effective suppression of pRSK with ERK1/2 inhibition (Morris et al.
2013 and Hatzivassiliou et al. 2012). Interestingly, BVD-523
treatment resulted in a marked increase in ERK1/2 phosphorylation
in vitro and in vivo. Similar to our findings, an increase in
pERK1/2 has been reported with the ERK1/2 inhibitor Vx11e;
conversely, pERK1/2 inhibition occurs with SCH772984 (Morris et al.
2013). Although differences in pERK1/2 levels were observed among
the various ERK1/2 inhibitors tested, downstream effectors (i.e.,
pRSK1 and total DUSP6) were similarly inhibited. These findings
suggest quantifying ERK1/2 target substrates, such as pRSK1, may
serve as reliable pharmacodynamic biomarkers for BVD-523-mediated
inhibition of ERK1/2 activity.
[0309] While BRAF (dabrafenib, vemurafenib) and MEK (trametinib,
cobimetinib) inhibitors validate the MAPK pathway as a therapeutic
target, particularly in patients with BRAF.sup.V600 mutations, the
antitumor response is limited by the emergence of acquired
resistance and subsequent disease progression. Resistance has been
attributed to the upregulation and activation of compensatory
signaling molecules (Nazarian et al. 2010, Villanueva et al. 2010,
Johannessen et al. 2010 and Wang et al. 2011), amplification of the
target genes (Corcoran et al. 2010), and activating mutations of
pathway components (e.g., RAS, MEK) (Wagle et al. 2011, Emery et
al. 2009 and Wang et al. 2011). Reactivation of the ERK1/2 pathway
is one common consequence of acquired resistance mechanism. When
introduced into the BRAF.sup.V600E-mutant melanoma cell line A375,
MEK.sup.Q56P conferred resistance to MEK and BRAF inhibition (Wagle
et al. 2011). By contrast, BVD-523 retained its potent inhibitory
activity in the engineered MEK.sup.Q56P cell line, indicating that
ERK1/2 inhibition is effective in the setting of upstream
activating alterations which can arise in response to BRAF/MEK
treatment. As further evidence of a role for BVD-523 in the context
of acquired resistance, efficacy of BVD-523 was evident in a
xenograft model derived from a tumor sample from a patient whose
disease progressed on vemurafenib; the BRAF inhibitor dabrafenib
was not effective in this model. These data support a role for
targeting ERK1/2 in the setting of BRAF/MEK resistance, and
complement previously published findings (Morris et al. 2013 and
Hatzivassiliou et al. 2012). To further characterize resistance to
inhibitors of the MAPK pathway, the emergence of resistance to
BVD-523 itself was investigated. It was found that single-agent
treatment of cancer cells with BVD-523 was durable and more
challenging to develop resistance compared with other agents
targeting upstream MAPK signaling components (i.e., dabrafenib,
trametinib). This may suggest that acquiring resistance to
ERK1/2-targeting agents is harder to achieve than acquiring
resistance to BRAF or MEK therapy, potentially due to the fact that
BVD-523 preferentially targets the more conserved active
confirmation of the ATP binding site. However, in vitro studies
with other ERK1/2 inhibitors have identified specific mutants in
ERK1/2 that drive resistance (Jha et al. 2016 and Goetz et al.
2014); these specific mutations have yet to be identified in
clinical samples from ERK1/2 inhibitor-relapsed patients.
[0310] The potential clinical benefit of ERK1/2 inhibition with
BVD-523 extends beyond the setting of BRAF/MEK therapy-resistant
patients. As ERK1/2 is a downstream master node within this MAPK
pathway, its inhibition is attractive in numerous cancer settings
where tumor growth depends on MAPK signaling. Approximately 30% of
all cancers harbor RAS mutations; therefore, targeting downstream
ERK1/2 with BVD-523 is a rational treatment approach for these
cancers. Furthermore, results from a study by Hayes et al. indicate
that prolonged ERK1/2 inhibition in KRAS-mutant pancreatic cancer
is associated with senescent-like growth suppression (Hayes et al.
2016). However, a combination approach may be required for maximal
and durable attenuation of MAPK signaling in the setting of RAS
mutations. For example, MEK inhibition in KRAS-mutant colorectal
cancer cell results in an adaptive response of ErbB family
activation, which dampens the response to MEK inhibition (Sun et
al. 2014). Similar context-specific adaptive responses may occur
following ERK1/2 inhibition with BVD-523. The optimal treatment
combinations for various genetic profiles and cancer histologies
are the subject of ongoing research. In addition to BRAF.sup.V600
and RAS mutations, other alterations which drive MAPK are emerging.
For example, novel RAF fusions and atypical non-V600 BRAF mutations
which promote RAF dimerization activate the MAPK pathway (Yao et
al. 2015). BRAF inhibitors such as vemurafenib and dabrafenib which
inhibit BRAFV600-mutant monomer proteins have been shown to be
inactive in atypical RAF alterations which drive MAPK signaling in
a dimerization-dependent manner (Yao et al. 2015). However,
treatment with BVD-523 to target downstream ERK1/2 in these tumors
may be a novel approach to addressing this unmet medical need.
[0311] In the setting of BRAF.sup.V600-mutant melanoma tumors,
combined BRAF and MEK inhibition exemplifies how agents targeting
different nodes of the same pathway can improve treatment response
and duration. Our combination studies in BRAF.sup.V600E-mutant
xenografts of human melanoma cell line A375 provides support for
combination therapy with BVD-523 and BRAF inhibitors. The
combination demonstrated superior benefit relative to single-agent
treatments, including results consistent with curative responses.
The clinical efficacy and tolerability of combined BRAF/BVD-523
therapy remains to be determined. It would not be unreasonable to
expect that a BRAF/ERK1/2 combination will at least be comparable
in efficacy to a targeted BRAF/MEK combination. Furthermore, the in
vitro observation that acquired resistance to BVD-523 is more
challenging to achieve compared with other MAPK pathway inhibitors
suggests that the BRAF/BVD-523 inhibitor combination has the
potential to provide a more durable response.
[0312] Significant progress has also been made using immunotherapy
for melanoma. The US FDA has approved various immune checkpoint
inhibitors for the treatment of advanced melanoma, including the
cytotoxic T-lymphocyte antigen-4 targeted agent ipilimumab and the
programmed death-1 inhibitors pembrolizumab and nivolumab.
Combining BVD-523 with such immunotherapies is an attractive
therapeutic option; further investigation is warranted to explore
dosing schedules and to assess whether synergistic response can be
achieved.
[0313] Based on the preclinical data, BVD-523 may hold promise for
treatment of patients with malignancies dependent on MAPK
signaling, including those whose tumors have acquired resistance to
other treatments. The clinical development of BVD-523 is described
below. See, Examples 17-24
Example 17
Phase I Dose-Escalation Study of the First-in-Class Novel Oral
ERK1/2 Kinase Inhibitor BVD-523 (Ulixertinib) in Patients with
Advanced Solid Tumors
[0314] The present invention describes the first-in-human dose
escalation study of an ERK1/2 inhibitor for the treatment of
patients with advanced solid tumors. BVD-523 has an acceptable
safety profile with favorable pharmacokinetics and early evidence
of clinical activity.
[0315] Mitogen-activated protein kinase (MAPK) signaling via the
RAS-RAF-MEK-ERK cascade plays a critical role in oncogenesis; thus
attracting significant interest as a therapeutic target. This
ubiquitous pathway is composed of RAS upstream of a cascade of the
protein kinases RAF, MEK1/2, and ERK1/2. RAS is activated by GTP
binding, which in turn results in activation of each protein kinase
sequentially. Although they appear to be the only physiologic
substrates for MEK1/2, ERK1/2 have many targets in the cytoplasm
and nucleus, including the transcription factors Elk1, c-Fos, p53,
Ets1/2, and c-Jun (Shaul et al. 2007). ERK1/2 activation and kinase
activity influences cellular proliferation, differentiation, and
survival through a variety of mechanisms (Rasola et al. 2010),
including activation of the ribosomal S6 kinase (RSK) family
members (Romeo et al. 2012).
[0316] Constitutive, aberrant activation of the
RAS-RAF-MEK1/2-ERK1/2 signaling pathway has been identified and
implicated in the development or maintenance of many cancers
(Schubbert et al. 2007 and Gollob et al. 2006). Mutations in RAS
family genes, such as KRAS, NRAS, and HRAS are the most common,
with activating RAS mutations occurring in .apprxeq.30% of human
cancers (Schubbert et al. 2007). KRAS mutations are prevalent in
pancreatic (>90%) (Kanda et al. 2012), biliary tract (3%-50%)
(Hezel et al. 2014), colorectal (30%-50%) (Arrington et al. 2012),
lung (27%) (Pennycuick et al. 2012), ovarian (15%-39%) (Dobrzycka
et al. 2009), and endometrioid endometrial (18%) (O'Hara and Bell
2012) cancers; NRAS mutations are prevalent in melanoma (20%)
(Khattak et al. 2013) and myeloid leukemia (8%-13%) (Yohe 2015);
and HRAS mutations are prevalent in bladder (12%) cancer
(Fernandez-Medarde and Santos 2011). Mutations in RAF family genes,
most notably BRAF, are frequent, particularly in melanoma. BRAF
mutations have been identified in 66% of malignant melanomas and in
.about.7% of a wide range of other cancers (Davies et al. 2002),
while MEK mutations are rarer, occurring at an overall frequency of
8% in melanomas (Nikolaev et al. 2012). In contrast, ERK mutations
resulting in tumorigenesis have been reported only rarely to date
(Deschenes-Simard et al. 2014).
[0317] The US Food and Drug Administration (FDA) has approved two
selective BRAF inhibitors, vemurafenib and dabrafenib, as
monotherapies for patients with BRAF.sup.V600-mutant metastatic
melanoma (Taflinar [package insert] and Zelboraf [package insert]).
Though response rates for these targeted therapies can be as high
as 50% in in patients with BRAF.sup.V600 mutations, duration of
response is often measured in months, not years (Hauschild et al.
2012 and McArthur et al. 2014). The MEK1/2 inhibitor trametinib is
also approved as a monotherapy in this setting (Mekinist [package
insert]), but is more commonly used in combination with the BRAF
inhibitor dabrafenib. First-line use of trametinib administered in
combination with dabrafenib offers an even greater improvement in
overall survival compared with vemurafenib monotherapy without
increased overall toxicity (Robert et al. 2015), highlighting the
potential utility of simultaneously targeting multiple proteins of
this MAPK signaling pathway. This therapeutic combination was also
associated with a lower incidence of MEK inhibitor-associated rash
and BRAF inhibitor-induced hyperproliferative skin lesions compared
with each single agent alone (Flaherty et al. 2012). Recently, a
phase III trial also demonstrated significant improvements in
overall survival (25.1 vs. 18.7 months, hazard ratio [HR]0.71,
P=0.0107), progression-free survival (PFS) (11.0 vs. 8.8 months, HR
0.67, P=0.0004), and overall response (69% vs. 53%; P=0.0014) with
dabrafenib plus trametinib versus dabrafenib alone in patients with
BRAF.sup.V600E/K mutation-positive melanoma (Long et al. 2015).
Similarly, significant improvements in PFS (9.9 vs. 6.2 months, HR
0.51, P<0.001) and the rate of complete response (CR) or partial
response (PR) (68% vs. 45%; P<0.001) have been demonstrated with
the combination of cobimetinib plus vemurafenib compared with
vemurafenib alone (Larkin et al. 2014). To this end, FDA approval
was recently granted for the combination of vemurafenib and
cobemetinib for BRAF.sup.V600E/K-mutated melanoma. Based on these
and related findings, the combination of a BRAF inhibitor plus a
MEK inhibitor has become a standard targeted treatment option for
patients with metastatic melanoma containing BRAF.sup.V600E/K
mutations.
[0318] Though BRAF/MEK-targeted combination therapy has been
demonstrated to provide significant additional benefit beyond
single-agent options, most patients eventually develop resistance
and disease progression after .about.12 months (Robert et al. 2015,
Flaherty et al. 2012 and Long et al. 2015). Several mechanisms of
acquired resistance following either single-agent or combination
therapies have been identified, including the generation of BRAF
splicing variants, BRAF amplification, development of NRAS or MEK
mutations, and upregulation of bypass pathways (Poulikakos et al.
2011, Corcoran et al. 2010, Nazarian et al. 2010, Shi et al. 2014,
Johannessen et al. 2010, Wagle et al. 2011, Wagle et al. 2014 and
Ahronian et al. 2015). Central to many of these mechanisms of
resistance is the reactivation of ERK signaling, which enables the
rapid recovery of MAPK pathway signaling and escape of tumor cells
from single-agent BRAF or combination BRAF/MEK inhibitor therapies
(Paraiso et al. 2010). ERK inhibition may provide the opportunity
to avoid or overcome resistance from upstream mechanisms, as it is
the most distal master kinase of this MAPK signaling pathway. This
is supported by preclinical evidence that inhibition of ERK by
small molecule inhibitors acted to both inhibit the emergence of
resistance and overcome acquired resistance to BRAF and MEK
inhibitors (Morris et al. 2013 and Hatzivassiliou et al. 2012).
[0319] BVD-523 is a highly potent, selective, reversible,
ATP-competitive ERK1/2 inhibitor which has been shown to reduce
tumor growth and induce tumor regression in BRAF and RAS mutant
xenograft models. Furthermore, single-agent BVD-523 inhibited human
xenograft models that were cross-resistant to both BRAF and MEK
inhibitors. See, Examples 9-16. Therefore, an open-label,
first-in-human study (Clinicaltrials.gov identifier, NCT01781429)
of oral BVD-523 to identify both the maximum tolerated dose and the
recommended dose for further study was undertaken. The present
study also aimed to assess pharmacokinetic and pharmacodynamic
properties as well as preliminary efficacy in patients with
advanced cancers.
Example 18
Patient Characteristics
[0320] A total of 27 patients were enrolled and received at least
one dose of study drug from Apr. 4, 2013 to Dec. 1, 2015. Baseline
demographics and disease characteristics are shown in Table 25. The
median patient age was 61 years (range, 33-86 years). Fifty-two
percent (14/27) of patients were male and 63% (17/27) had an
Eastern Cooperative Oncology Group (ECOG) performance status of 1.
Melanoma was the most common cancer (30%; BRAF mutation present in
7/8 of these patients). The remaining patients had colorectal (19%;
5/27), papillary thyroid (15%; 4/27), or non-small cell lung cancer
(NSCLC) (7%; 2/27), and 8 (30%) were classified as having other
cancers (2 pancreatic, 1 appendiceal, 1 nonseminomatous germ cell,
1 ovarian and 3 with unknown primary). The majority of patients had
received 2 or more prior lines of systemic therapy, with 41%
(11/27) receiving 2 to 3 and 48% (13/27) receiving >3 prior
lines of systemic therapy.
TABLE-US-00025 TABLE 25 Baseline demographics and clinical
characteristics of patients Parameter N = 27 Median age, years
(range) 61 (33-86) Sex, n (%) Female 13 (48) Male 14 (52)
Ethnicity, n (%) Not Hispanic/Latino 27 (100) ECOG performance
status 0 10 (37) 1 17 (63) Cancer type, n (%) Melanoma.sup.a 8 (30)
Colorectal 5 (19) Papillary thyroid 4 (15) Non-small cell lung 2
(7) Other.sup.b 8 (30) Molecular abnormalities, n (%).sup.c BRAF
mutant 13 (48) KRAS mutant 6 (22) NRAS mutant 2 (7) Other.sup.d 7
(26) Unknown 4 (15) Number of prior systemic anticancer regimens, n
(%) 0 1 (4) 1 2 (7) 2-3 11 (41) >3 13 (48) Prior
BRAF/MEK-targeted therapy.sup.e, n (%) 11 (41) BRAF 5 (19) MEK 6
(22) BRAF/MEK 2 (7) .sup.aSeven were BRAF mutant and 1 was unknown.
.sup.bTwo pancreatic, 1 appendiceal, 1 non-seminomatous germ cell,
1 ovarian, 3 unknown primary. .sup.cPatients may have more than 1
molecular abnormality. .sup.dOther molecular abnormalities included
ERCC1, RRM1, thymidylate synthetase, GNAS, MEK1, TP53, CREBBP,
ROS1, PTEN, AKT3, and PIK3CA. .sup.eSome patients were treated with
more than one BRAF inhibitor. Abbreviation: ECOG, Eastern
Cooperative Oncology Group.
Example 19
Ex Vivo Effects of BVD-523 on RSK1/2 Phosphorylation
[0321] An ex vivo biomarker assay that could be used to support
clinical studies was developed to demonstrate the inhibitory
effects of BVD-523 on ERK activity. The assay extends preclinical
cellular data where inhibitors of MAPK signaling, such as BVD-523,
dabrafenib, trametinib, and vemurafenib, have been shown to inhibit
RSK phosphorylation as a function of inhibitor concentration in
BRAF mutant cancer cell lines. See, Examples 9-16. Specifically,
ERK inhibitor-dependent inhibition of phorbol 12-myristate
13-acetate (PMA)-stimulated phosphorylation of the ERK substrate
RSK1 in whole blood was used as a target marker. When BVD-523 was
added directly to whole blood from healthy volunteers,
PMA-stimulated RSK phosphorylation decreased with increasing
concentrations of BVD-523 (FIG. 38). The mean IC.sub.50 for the
cumulative data was 461.+-.20 nM for BVD-523, with a maximum
inhibition of 75.8.+-.2.7% at 10 .mu.M BVD-523. Maximum inhibition
was defined as the RSK phosphorylation measured in the presence of
10 .mu.M BVD-523. Patient-derived whole blood samples, collected
just prior to dosing or at defined timepoints following dosing with
BVD-523, were similarly treated and RSK phosphorylation levels
quantitated.
Example 20
Dose Escalation, Dose-Limiting Toxicities (DLTs), Maximum Tolerated
Dose (MTD), and Recommended Phase II Dose (RP2D)
[0322] As per protocol, 5 single-patient cohorts (from 10 to 150 mg
twice-daily [BID]) proceeded without evidence of a DLT. The 300-mg
BID cohort was expanded to more fully characterize BVD-523
exposures. One of 6 patients given 600 mg BID experienced a DLT of
Grade 3 rash. The 900-mg BID dose exceeded the MTD, with one
patient experiencing Grade 3 pruritus and elevated aspartate
aminotransferase (AST) and another patient experiencing Grade 3
diarrhea, vomiting, dehydration, and elevated creatinine (Table
26). The subsequent intermediate dose of 750 mg BID also exceeded
the MTD, with DLTs of Grade 3 rash and Grade 2 diarrhea in 1
patient and Grade 2 hypotension, elevated creatinine, and anemia in
another patient. Therefore, the MTD and RP2D were determined to be
600 mg BID.
TABLE-US-00026 TABLE 26 Dose-limiting toxicities in Cycle 1 (21
days) Dose, mg DLT (BID) Frequency DLT Description 10 0/1 N/A 20
0/1 N/A 40 0/1 N/A 75 0/1 N/A 150 0/1 N/A 300 0/4 N/A 600 1/8 Rash
(Grade 3) 750.sup.a 2/4 Rash (Grade 3), diarrhea (Grade 2)
Hypotension (Grade 2), elevated creatinine (Grade 2), anemia (Grade
2), delay to cycle 2 dosing 900 2/7 Pruritus (Grade 3), elevated
AST (Grade 3) Diarrhea (Grade 3), vomiting (Grade 3), dehydration
(Grade 3), elevated creatinine (Grade 3) .sup.aIntermediate dose.
Abbreviations: AST, aspartate transaminase, BID, twice daily; DLT,
dose-limiting toxicity; N/A, not applicable.
Example 21
Adverse Events (AEs)
[0323] Investigator-assessed treatment-related AEs of any grade
were noted in 26 of 27 patients (96%). The most common
treatment-related AEs (>30%) were rash (predominately acneiform)
(70%), fatigue (59%), diarrhea (52%), and nausea (52%) (Table 27).
No patients experienced a Grade 4 or 5 treatment-related AE or
discontinued treatment due to a treatment-related AE. Most events
were Grade 1 to 2, with treatment-related Grade 3 events noted in
13 of 27 patients (48%). The only Grade 3 treatment-related events
present in >10% of patients were diarrhea (15%) and increased
liver function tests (11%), all of which occurred above the 600-mg
BID dose.
TABLE-US-00027 TABLE 27 Adverse events possibly/definitely related
to BVD-523 in .gtoreq.10% of patients N = 27 Any grade, Grade 1 or
2, Grade 3.sup.a, Event n (%) n (%) n (%) Rash 20 (74) 18 (67)
2.sup.b (7).sup. Fatigue 17 (63) 16 (59) 1 (4) Diarrhea 16 (59) 12
(44) 4 (15) Nausea 14 (52) 14 (52) 0 Vomiting 8 (30) 7 (26) 1 (4)
Anorexia 6 (22) 6 (22) 0 Pruritus 6 (22) 6 (22) 0 Anemia 5 (19) 3
(11) 2 (7) Increased creatinine 5 (19) 4 (15) 1 (4) Dehydration 5
(19) 3 (11) 2 (7) Peripheral edema 4 (15) 4 (15) 0 Increased LFTs 4
(14) 1 (4) 3 (11) (ALT and AST) Blurry/dimmed vision.sup.c 3 (11) 3
(11) 0 Constipation 3 (11) 3 (11) 0 Fever 3 (11) 3 (11) 0 .sup.aNo
patients experienced Grade 4 or 5 AEs that were possibly or
definitely related to BVD-523 treatment. .sup.bAcneiform and
maculo-papular rash. .sup.cOne Grade 1 event of related central
serous retinopathy. Analysis cut-off date: Dec. 1, 2015.
Abbreviations: AEs, adverse events; ALT, alanine transaminase; AST,
aspartate transaminase; LFTs, liver function tests.
[0324] Fourteen patients experienced a total of 28 serious AEs
(SAEs). Nine of these were considered to be related or possibly
related to BVD-523 by the investigator, which included dehydration,
diarrhea, or elevated creatinine (2 patients each), vomiting,
nausea, and fever (1 patient each). All other SAEs were considered
to be unrelated to treatment with BVD-523. Dose reductions
resulting from AEs occurred in 3 patients during the study: 1
patient reduced from 600 mg BID to 300 mg BID and 2 patients
reduced from 900 mg BID to 600 mg BID.
Example 22
Pharmacokinetics
[0325] Single-dose and steady-state pharmacokinetics of BVD-523 are
summarized in FIG. 39A and Table 28. Generally, orally administered
BVD-523 was slowly absorbed in patients with advanced malignancies.
After reaching the maximum concentration (C.sub.max), plasma
BVD-523 levels remained sustained for approximately 2 to 4 hours.
Subsequently, plasma drug concentrations slowly declined. Since
plasma drug concentrations were measured only up to 12 hours after
the morning dose, it was not possible to calculate an effective or
terminal phase elimination rate. BVD-523 pharmacokinetics were
linear and dose proportional in terms of both C.sub.max and area
under the curve (AUC) when administered up to 600 mg BID. A further
increase in exposure was not observed as the dose increased from
600 to 900 mg BID. The C.sub.max reached the level of the EC.sub.50
based on the ex vivo whole blood assay (.apprxeq.200 ng/mL) for all
doses above 20 mg BID. Additionally, steady-state exposures
remained at or above the target EC.sub.50 for dose levels of
.gtoreq.150 mg BID throughout the dosing period. Minimal plasma
accumulation of BVD-523 and its metabolites were observed on Day 15
at the lower (<75 mg BID) dose levels, whereas accumulation
ranged from approximately 1.3- to 4.0-fold for the higher dose
levels. Predose concentrations on Day 22 were generally similar to
those on Day 15, indicating that steady state had already been
attained by Day 15 (data not shown). The degree of interpatient
variability in plasma exposure to BVD-523 and its metabolites was
considered moderate and not problematic.
TABLE-US-00028 TABLE 28 Steady-state BVD-523 pharmacokinetics
(Cycle 1, Day 15) C.sub.max, ng/mL .+-. SD AUC.sub.0-22, ng hr/mL
.+-. SD Dose, mg.sup.a n.infin. Day 1 Day 15 Day 1 Day 15 10 1 48.2
45.7 220 234 20 1 14.9 15.8 91.7 98.7 40 1 100 191 614 999 150 1
133 326 817 2770 300 4.sup.b 765 .+-. 234 586 .+-. 257 4110 .+-.
1140 4460 .+-. 2460 600.sup.c 7.sup.d 1110 .+-. 589 2750 .+-. 1740
2750 .+-. 1740 24400 .+-. 16200 750 4.sup.b 1450 .+-. 539 2290 .+-.
1790.sup.f 10700 .+-. 1120.sup.g 23300 .+-. 19800.sup.f 900 7.sup.e
1430 .+-. 1010 1720 .+-. 328 10800 .+-. 6320.sup.h 15900 .+-.
1300.sup.g .sup.aDose level administered twice daily; .sup.bn = 3
on Day 15; .sup.cNumber of subjects for Day 15 at the 600 mg dose
level includes two subjects who started Day 1 dosing at 900 mg and
were later reduced to 600 mg; .sup.dn = 8 on Day 15; .sup.en = 4 on
Day 15; .sup.fOne subject started on Day 1 dosing at 750 mg and was
later reduced to 450 mg. Day 15 parameters for this subject reflect
at least 10 consecutive doses at 450 mg/dose. Individual Day 15
parameters were 1300 ng/mL for C.sub.max and 10700 ng hr/mL for
AU.sub.max; .sup.gn = 3; .sup.hn = 5.
[0326] The urinary excretion after first dose and at steady state
of BVD-523 was negligible (<0.2% of the dose) at all dose levels
within 12 hours postdose, and not dose-related within this very low
percentage range. Renal clearance appeared to be dose-independent.
Individual renal clearance values ranged from 0.128 to 0.0895 L/hr
(where n=1 per dose level) and mean values ranged from 0.0149 to
0.0300 L/hr (where n.gtoreq.3).
Example 23
Pharmacodynamic Confirmation of Target Inhibition by BVD-523
[0327] To confirm on-target and pathway inhibition by BVD-523,
RSK-1 phosphorylation was examined as a target biomarker in human
whole blood samples from patients with solid tumors who received
BVD-523. Steady state whole blood samples collected just prior to
Day 15 dosing from BVD-523-treated patients displayed
concentration-dependent inhibition of PMA stimulated ERK activity
(FIG. 39B), ranging from 0% ERK inhibition with BVD-523 dosing at
10 mg BID to 93.+-.8% ERK inhibition with dosing at 900 mg BID. The
plasma concentrations of BVD-523 that yielded 50% inhibition of ERK
phosphorylation were similar whether BVD-523 was spiked directly
into healthy volunteer plasma or was present following oral dosing
of patients.
Example 24
Antitumor Effects
[0328] Tumor response to BVD-523 was assessed in 25 evaluable
patients using Response Evaluation Criteria in Solid Tumors version
1.1 (RECIST v1.1); 2 patients did not receive both scans of target
lesions and were thus not evaluated using RECIST v1.1. No patients
achieved a complete response, but 3 patients (all patients with
melanoma with BRAF.sup.V600 mutations) achieved a partial response
(129 days [BRAF/MEK-inhibitor naive], 294 days ongoing at
[refractory to prior BRAF/MEK inhibitors], 313 days ongoing by the
data cutoff date [intolerant to other BRAF/MEK inhibitors]) (FIG.
40A). Interestingly, all 3 partial responders had BRAF-mutant
melanoma. One partial responder, who was receiving BVD-523 at a
dose of 450 mg BID, had an approximate 70% reduction in the sum of
target lesions from baseline, while the other partial responders
showed reductions of 47.0% and 33.6%. Stable disease was
demonstrated in 18 patients, with 6 having stable disease for more
than 6 months, and 6 additional patients having stable disease for
more than 3 months. In this study, 4 patients displayed progressive
disease at first evaluation.
[0329] FIG. 40B shows computed tomography (CT) scans of 1 of the 3
partial responders (RECIST v1.1) who had progressed on prior
vemurafenib and subsequent dabrafenib/trametinib treatment; a
durable partial response was observed following dosing of BVD-523
600 mg BID for >300 days. BVD-523 was associated with a
metabolic response using fluorodeoxyglucose-positive emission
tomography (.sup.18F-FDG-PET) in 5 of 16 evaluable patients.
[0330] FIG. 41 depicts the time to response and the duration of
response in the study population. The two patients who demonstrated
responses to BVD-523 remained on study and continued with BVD-523
treatment as of the study cutoff date (>500 days); additionally,
one patient with bronchoalveolar NSCLC (not enough tissue for
molecular profiling) had been on treatment for >700 days with
stable disease. Twenty-four of 27 patients (90%) discontinued
treatment due to progressive disease (22/27, 82%) or other reasons
(2/27, 7%). The mean duration of BVD-523 treatment before
discontinuation was 4.7 months.
Discussion
[0331] The present invention presents results from a first-in-human
study evaluating the safety, pharmacokinetics, pharmacodynamics,
and preliminary efficacy of BVD-523 in 27 patients with advanced
solid tumors. In this dose-escalation study, oral treatment with
BVD-523 resulted in both radiographic responses by RECIST v1.1 (3
partial responses) and prolonged disease stabilization in some
patients, the majority of whom had been treated with 22 prior
systemic therapies. Evidence of BVD-523-dependent inhibition of
metabolic response in tumors was established in a subset of
patients by imaging tumor uptake of .sup.18F-glucose. Drug
exposures increased linearly with increasing doses up to 600 mg
BID, with exposures at 600 mg BID providing near complete 24/7
inhibition of ERK-dependent substrate (RSK-1) phosphorylation in an
ex vivo whole blood assay. Furthermore, tolerability to BVD-523 was
manageable when administered up to its MTD and RP2D, determined to
be 600 mg BID.
[0332] BVD-523 was generally well tolerated, with manageable and
reversible toxicity. The most common AEs were rash (usually
acneiform), fatigue, and gastrointestinal side effects, including
nausea, vomiting, and diarrhea. The safety profile of BVD-523 is
consistent with its selective inhibition of the MAPK pathway; the
AE profile shows considerable overlap with MEK inhibitor
experience. However, toxicities associated with any targeted
therapy may include dependence on both the specific mechanism and
the degree of target inhibition as well as any off-target effects
(Zelboraf [package insert] and Hauschild et al. 2012). Ongoing and
future investigations will extend both the efficacy and safety
profile demonstrated in this dose-escalation study, and will guide
how the unique profile of the ERK inhibitor BVD-523 might be used
as a single agent or in combination with other agents.
[0333] Durable responses by RAF and MEK inhibitors are often
limited by intrinsic and eventual acquired resistance, with a
common feature often involving reactivation of the ERK pathway
(Poulikakos et al. 2011, Corcoran et al. 2010, Nazarian et al.
2010, Shi et al. 2014, Johannessen et al. 2010, Wagle et al. 2011,
Wagle et al. 2014, Ahronian et al. 2015 and Paraiso et al. 2010).
Thus, ERK inhibition with BVD-523 alone or in combination with
other MAPK signaling pathway inhibitors may have the potential to
delay the development of resistance to existing therapies and to
benefit a broader patient population. That ERK inhibitors,
including BVD-523, retain their potency in BRAF- and MEK-resistant
cell lines provide preclinical evidence for the use of ERK
inhibitors in patients with acquired resistance to standard of care
(BRAF/MEK combination therapy) See, e.g., Examples 9-16.
Importantly, in this study, a patient whose cancer had progressed
after experiencing stable disease when treated initially with a
BRAF inhibitor (vemurafenib) and subsequently with a combination of
BRAF and MEK inhibitors (dabrafenib/trametinib) had a partial
response when receiving single-agent BVD-523. This patient has
remained on-study for a total of 708 days, as of the cutoff date of
the study reported herein. Based in part on the antitumor effects
observed in this patient, the FDA has designated as a Fast Track
development program the investigation of BVD-523 for the treatment
of patients with unresectable or metastatic BRAF.sup.V600
mutation-positive melanoma that is refractory to or has progressed
following treatment with a BRAF and/or MEK inhibitor(s). Precise
definition of exactly how BVD-523 might best support patient care
(eg, as a single agent or in various combinations) requires
additional clinical studies.
[0334] In summary, the present examples present data from an
initial data from the dose escalation portion of a phase I study
evaluating BVD-523, a novel first-in-class ERK inhibitor, as a
treatment for patients with advanced cancers. Continuous,
twice-daily oral treatment with BVD-523 resulted in antitumor
effects in several patients, including patients either naive to or
having progressed on available MAPK pathway-targeted therapies.
BVD-523 was generally well tolerated in this advanced cancer
patient population and toxicities were manageable; the MTD and RP2D
were 600 mg BID. BVD-523 exposures increased linearly up to the
RP2D and robust pharmacodynamics effects were evident at this dose
level. An expansion of this phase I clinical study is currently
underway to confirm and extend the observations made in the
dose-escalation phase. Specifically, patients are being enrolled
into molecularly classified expansion cohorts (e.g., NRAS, BRAF,
MEK or ERK alterations) across various tumor histologies.
Furthermore, expansion cohorts are evaluating the use of BVD-523 in
patients with cancer who are either naive to available MAPK pathway
therapies or those whose disease has progressed on such
treatments.
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[0417] All documents cited in this application are hereby
incorporated by reference as if recited in full herein.
[0418] Although illustrative embodiments of the present invention
have been described herein, it should be understood that the
invention is not limited to those described, and that various other
changes or modifications may be made by one skilled in the art
without departing from the scope or spirit of the invention.
Sequence CWU 1
1
8212949DNAHomo sapiens 1cgcctccctt ccccctcccc gcccgacagc ggccgctcgg
gccccggctc tcggttataa 60gatggcggcg ctgagcggtg gcggtggtgg cggcgcggag
ccgggccagg ctctgttcaa 120cggggacatg gagcccgagg ccggcgccgg
cgccggcgcc gcggcctctt cggctgcgga 180ccctgccatt ccggaggagg
tgtggaatat caaacaaatg attaagttga cacaggaaca 240tatagaggcc
ctattggaca aatttggtgg ggagcataat ccaccatcaa tatatctgga
300ggcctatgaa gaatacacca gcaagctaga tgcactccaa caaagagaac
aacagttatt 360ggaatctctg gggaacggaa ctgatttttc tgtttctagc
tctgcatcaa tggataccgt 420tacatcttct tcctcttcta gcctttcagt
gctaccttca tctctttcag tttttcaaaa 480tcccacagat gtggcacgga
gcaaccccaa gtcaccacaa aaacctatcg ttagagtctt 540cctgcccaac
aaacagagga cagtggtacc tgcaaggtgt ggagttacag tccgagacag
600tctaaagaaa gcactgatga tgagaggtct aatcccagag tgctgtgctg
tttacagaat 660tcaggatgga gagaagaaac caattggttg ggacactgat
atttcctggc ttactggaga 720agaattgcat gtggaagtgt tggagaatgt
tccacttaca acacacaact ttgtacgaaa 780aacgtttttc accttagcat
tttgtgactt ttgtcgaaag ctgcttttcc agggtttccg 840ctgtcaaaca
tgtggttata aatttcacca gcgttgtagt acagaagttc cactgatgtg
900tgttaattat gaccaacttg atttgctgtt tgtctccaag ttctttgaac
accacccaat 960accacaggaa gaggcgtcct tagcagagac tgccctaaca
tctggatcat ccccttccgc 1020acccgcctcg gactctattg ggccccaaat
tctcaccagt ccgtctcctt caaaatccat 1080tccaattcca cagcccttcc
gaccagcaga tgaagatcat cgaaatcaat ttgggcaacg 1140agaccgatcc
tcatcagctc ccaatgtgca tataaacaca atagaacctg tcaatattga
1200tgacttgatt agagaccaag gatttcgtgg tgatggagga tcaaccacag
gtttgtctgc 1260taccccccct gcctcattac ctggctcact aactaacgtg
aaagccttac agaaatctcc 1320aggacctcag cgagaaagga agtcatcttc
atcctcagaa gacaggaatc gaatgaaaac 1380acttggtaga cgggactcga
gtgatgattg ggagattcct gatgggcaga ttacagtggg 1440acaaagaatt
ggatctggat catttggaac agtctacaag ggaaagtggc atggtgatgt
1500ggcagtgaaa atgttgaatg tgacagcacc tacacctcag cagttacaag
ccttcaaaaa 1560tgaagtagga gtactcagga aaacacgaca tgtgaatatc
ctactcttca tgggctattc 1620cacaaagcca caactggcta ttgttaccca
gtggtgtgag ggctccagct tgtatcacca 1680tctccatatc attgagacca
aatttgagat gatcaaactt atagatattg cacgacagac 1740tgcacagggc
atggattact tacacgccaa gtcaatcatc cacagagacc tcaagagtaa
1800taatatattt cttcatgaag acctcacagt aaaaataggt gattttggtc
tagctacagt 1860gaaatctcga tggagtgggt cccatcagtt tgaacagttg
tctggatcca ttttgtggat 1920ggcaccagaa gtcatcagaa tgcaagataa
aaatccatac agctttcagt cagatgtata 1980tgcatttgga attgttctgt
atgaattgat gactggacag ttaccttatt caaacatcaa 2040caacagggac
cagataattt ttatggtggg acgaggatac ctgtctccag atctcagtaa
2100ggtacggagt aactgtccaa aagccatgaa gagattaatg gcagagtgcc
tcaaaaagaa 2160aagagatgag agaccactct ttccccaaat tctcgcctct
attgagctgc tggcccgctc 2220attgccaaaa attcaccgca gtgcatcaga
accctccttg aatcgggctg gtttccaaac 2280agaggatttt agtctatatg
cttgtgcttc tccaaaaaca cccatccagg cagggggata 2340tggtgcgttt
cctgtccact gaaacaaatg agtgagagag ttcaggagag tagcaacaaa
2400aggaaaataa atgaacatat gtttgcttat atgttaaatt gaataaaata
ctctcttttt 2460ttttaaggtg aaccaaagaa cacttgtgtg gttaaagact
agatataatt tttccccaaa 2520ctaaaattta tacttaacat tggattttta
acatccaagg gttaaaatac atagacattg 2580ctaaaaattg gcagagcctc
ttctagaggc tttactttct gttccgggtt tgtatcattc 2640acttggttat
tttaagtagt aaacttcagt ttctcatgca acttttgttg ccagctatca
2700catgtccact agggactcca gaagaagacc ctacctatgc ctgtgtttgc
aggtgagaag 2760ttggcagtcg gttagcctgg gttagataag gcaaactgaa
cagatctaat ttaggaagtc 2820agtagaattt aataattcta ttattattct
taataatttt tctataacta tttcttttta 2880taacaatttg gaaaatgtgg
atgtctttta tttccttgaa gcaataaact aagtttcttt 2940ttataaaaa
29492765PRTHomo sapiens 2Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly
Ala Glu Pro Gly Gln Ala 1 5 10 15 Leu Phe Asn Gly Asp Met Glu Pro
Glu Ala Gly Ala Gly Ala Gly Ala 20 25 30 Ala Ala Ser Ser Ala Ala
Asp Pro Ala Ile Pro Glu Glu Val Trp Asn 35 40 45 Ile Lys Gln Met
Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu 50 55 60 Asp Lys
Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala 65 70 75 80
Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln 85
90 95 Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser
Ser 100 105 110 Ser Ala Ser Met Asp Thr Val Thr Ser Ser Ser Ser Ser
Ser Leu Ser 115 120 125 Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn
Pro Thr Asp Val Ala 130 135 140 Arg Ser Asn Pro Lys Ser Pro Gln Lys
Pro Ile Val Arg Val Phe Leu 145 150 155 160 Pro Asn Lys Gln Arg Thr
Val Val Pro Ala Arg Cys Gly Val Thr Val 165 170 175 Arg Asp Ser Leu
Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu 180 185 190 Cys Cys
Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly 195 200 205
Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu 210
215 220 Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys
Thr 225 230 235 240 Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys
Leu Leu Phe Gln 245 250 255 Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys
Phe His Gln Arg Cys Ser 260 265 270 Thr Glu Val Pro Leu Met Cys Val
Asn Tyr Asp Gln Leu Asp Leu Leu 275 280 285 Phe Val Ser Lys Phe Phe
Glu His His Pro Ile Pro Gln Glu Glu Ala 290 295 300 Ser Leu Ala Glu
Thr Ala Leu Thr Ser Gly Ser Ser Pro Ser Ala Pro 305 310 315 320 Ala
Ser Asp Ser Ile Gly Pro Gln Ile Leu Thr Ser Pro Ser Pro Ser 325 330
335 Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His
340 345 350 Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro
Asn Val 355 360 365 His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp
Leu Ile Arg Asp 370 375 380 Gln Gly Phe Arg Gly Asp Gly Gly Ser Thr
Thr Gly Leu Ser Ala Thr 385 390 395 400 Pro Pro Ala Ser Leu Pro Gly
Ser Leu Thr Asn Val Lys Ala Leu Gln 405 410 415 Lys Ser Pro Gly Pro
Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu 420 425 430 Asp Arg Asn
Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp 435 440 445 Trp
Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser 450 455
460 Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala
465 470 475 480 Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln
Leu Gln Ala 485 490 495 Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr
Arg His Val Asn Ile 500 505 510 Leu Leu Phe Met Gly Tyr Ser Thr Lys
Pro Gln Leu Ala Ile Val Thr 515 520 525 Gln Trp Cys Glu Gly Ser Ser
Leu Tyr His His Leu His Ile Ile Glu 530 535 540 Thr Lys Phe Glu Met
Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala 545 550 555 560 Gln Gly
Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu 565 570 575
Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly 580
585 590 Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His
Gln 595 600 605 Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro
Glu Val Ile 610 615 620 Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln
Ser Asp Val Tyr Ala 625 630 635 640 Phe Gly Ile Val Leu Tyr Glu Leu
Met Thr Gly Gln Leu Pro Tyr Ser 645 650 655 Asn Ile Asn Asn Arg Asp
Gln Ile Ile Phe Met Val Gly Arg Gly Tyr 660 665 670 Leu Ser Pro Asp
Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met 675 680 685 Lys Arg
Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro 690 695 700
Leu Phe Pro Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser Leu 705
710 715 720 Pro Lys Ile His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg
Ala Gly 725 730 735 Phe Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys Ala
Ser Pro Lys Thr 740 745 750 Pro Ile Gln Ala Gly Gly Tyr Gly Ala Phe
Pro Val His 755 760 765 3 3906DNARattus norvegicus 3atggcggcgc
tgagtggcgg cggtggcagc agcagcggtg gcggtggcgg cggcggcggc 60ggcggtggtg
gcggcggcgg cggcggcgcc gaacagggac aggctctgtt caatggcgac
120atggagccgg aggccggcgc tggcgccgcg gcctcttcgg ccgcggaccc
ggccattcct 180gaagaggtgt ggaatatcaa gcaaatgatt aagttgacac
aggaacatat agaggcccta 240ttggacaagt ttggtgggga gcataaccca
ccgtcaatat acctggaggc ctatgaagag 300tacaccagca agctagatgc
ccttcagcag agagagcagc agctgttgga atccctggtt 360tttcaaactc
ccacagatgt atcacggaac aaccccaagt caccacagaa acctatcgtt
420cgtgtcttcc tgcccaacaa acagaggaca gtggtgcccg caagatgtgg
tgtaacggtc 480cgagacagtc taaagaaagc actaatgatg aggggtctca
tcccagagtg ctgtgctgtt 540tacagaattc aggacggaga gaagaaacca
attggctggg acactgacat ttcctggctt 600actggagagg agctacatgt
tgaagtacta gagaatgttc ctctgacaac ccacaacttc 660gtacggaaaa
cttttttcac cttagcattt tgtgactttt gccgaaagct gcttttccag
720ggtttccgct gtcaaacatg tggttataag tttcaccagc gttgtagtac
agaggttcca 780ctgatgtgtg ttaattatga ccaacttgat ttgctgtttg
tctccaagtt ctttgagcat 840cacccagtac cacaggagga ggccttctca
gcagagacta cccttccatc tggatgctct 900tccgcacccc cctcagactc
tattgggccc caaatcctca ccagtccatc tccttcaaaa 960tccattccaa
ttccacagcc cttccggcca gcagatgaag atcatcgcaa tcagtttggg
1020caacgagacc gctcctcctc cgctcccaat gttcatataa acacaatcga
acctgtcaat 1080attgatgaaa aattcccaga agtggaatta caggatcaaa
gggatttgat tagagaccag 1140gggtttcgtg gggatggagc ccctttgaac
cagctgatgc gctgtcttcg gaaataccaa 1200tcccggactc ccagccccct
cctccattct gtccccagtg aaatagtgtt tgattttgag 1260cctggcccag
tgttcagagg gtcaaccaca ggcttgtcgg ccaccccacc tgcctcatta
1320cctggctcac tcactaacgt gaaagcctta cagaaatctc caggacctca
gcgggaaagg 1380aagtcctcct cctcctcctc ctccacggaa gacagaagtc
ggatgaaaac acttggtaga 1440agagattcaa gtgatgattg ggagattcct
gatggacaga ttacagtggg acagagaatt 1500ggatctgggt cctttggaac
tgtctacaag ggaaagtggc atggcgacgt ggcagtgaaa 1560atgctgaatg
tgacagcacc cacacctcag cagttacagg ccttcaaaaa cgaagtcgga
1620gtactcagga aaactcgaca tgtgaacatc ctccttttca tgggctattc
tacaaagcca 1680cagctggcta ttgttacaca gtggtgtgaa ggctccagct
tatatcacca tctccacatc 1740attgagacca aatttgagat gatcaaactt
atagatattg cacggcagac tgcacagggc 1800atggattact tacacgccaa
gtcaatcatc cacagagacc tcaagagtaa taatatattt 1860cttcatgaag
acctcacggt aaaaataggt gactttggtt tagccacagt gaagtcccga
1920tggagtgggt cccatcagtt tgaacagttg tctggatcta ttttgtggat
ggcacccgaa 1980gtaatcagaa tgcaagataa aaacccatat agctttcagt
cagacgtgta tgcatttggg 2040attgttctgt atgaactgat gactggtcag
ctaccttatt caaacatcaa caacagggat 2100cagataattt ttatggtggg
acgaggatac ctatctccag atctcagtaa ggtacggagt 2160aactgtccaa
aagccatgaa gagattaatg gcagagtgcc tcaaaaagaa aagagacgag
2220agaccactct ttccccaaat tctcgcctct attgagctgc tggcccgctc
attgccaaaa 2280attcaccgca gtgcatcaga accctccttg aatcgggctg
gtttccaaac agaagatttt 2340agtctgtatg cttgtgcttc tccaaaaaca
cccatccaag cagggggata tggagaattt 2400gcagccttca agtagccact
ccatcatggc agcatctact ctttatttct taagtcttgt 2460gttcatacaa
tttgttaaca tcaaaacaca gttctgttcc tcaaattttt tttaaagata
2520caaaattttc aatgcataag ctcgtgtgga acagaatgga atttcctatt
caacaaaaga 2580gggaagaatg ttttaggaac cagaattctc tgctgcccgt
gtttcttctt caacacaaat 2640atcatgtgca tacaactctg cccattccca
agaagaaaga ggagagaccc cgaattctgc 2700ccttttggtg gtcaggcatg
atggaaagaa tttgctgctg cagcttggga aaaattgcta 2760tggaaagtct
gccagtcaac tttgcccttc taaccaccag atccatttgt ggctggtcat
2820ctgatggggc gatttcaatc accaagcatc gttcttgcct gttgtgggat
tatgtcgtgg 2880agcactttcc ctatccacca ccgttaattt ccgagggatg
gagtaaatgc agcataccct 2940ttgtgtagca cctgtccagt cctcaaccaa
tgctatcaca gtgaagctct ttaaatttaa 3000gtggtgggtg agtgttgagg
agagactgcc ttgggggcag agaaaagggg atgctgcatc 3060ttcttcctca
cctccagctc tctcacctcg ggttgccttg cacactgggc tccgcctaac
3120cactcgggct gggcagtgct ggcacacatt gccgcctttt ctcattgggt
ccagcaattg 3180agcagagggt tgggggattg tttcctccac aatgtagcaa
attctcagga aaatacagtc 3240catatcttcc tctcagctct tccagtcacc
aaatacttac gtggctcctt tgtccaggac 3300ataaaacacc gtggacaaca
cctaattaaa agcctacaaa actgcttact gacagttttg 3360aatgtgagac
atttgtgtaa tttaaatgta aggtacaggt cttaatttct tctattaagt
3420ttcttctatt tttatttaaa cgaagaaaat aattttcagg tttaattgga
ataaacgaat 3480acttcccaaa agactatata ccctgaaaat tatatttttg
ttaattgtaa acaactttta 3540aaaaatggtt attatccttt tctctaccta
aaattatggg aaatcttagc ataatgacaa 3600ttatttatac tttttaaata
aatggtactt gctggatcca cactaacatc tttgctaaca 3660ttcccattgt
ttcttccaac ttcactccta cactacatcc tccatcctct ttctagtctt
3720ttatctataa tatgcaacct aaaataaaag tggtggtgtc tccattcatt
cttcttcttc 3780cttttttccc caagcctggt cttcaaaagg ttgggtaatt
tagtagctga gttccctagg 3840tagaaataga actattaggg acattggggt
tgtaggaaag cgtgaggcct gtcaccagtt 3900gttctt 39064804PRTRattus
norvegicus 4Met Ala Ala Leu Ser Gly Gly Gly Gly Ser Ser Ser Gly Gly
Gly Gly 1 5 10 15 Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly
Gly Ala Glu Gln 20 25 30 Gly Gln Ala Leu Phe Asn Gly Asp Met Glu
Pro Glu Ala Gly Ala Gly 35 40 45 Ala Ala Ala Ser Ser Ala Ala Asp
Pro Ala Ile Pro Glu Glu Val Trp 50 55 60 Asn Ile Lys Gln Met Ile
Lys Leu Thr Gln Glu His Ile Glu Ala Leu 65 70 75 80 Leu Asp Lys Phe
Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu 85 90 95 Ala Tyr
Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu 100 105 110
Gln Gln Leu Leu Glu Ser Leu Val Phe Gln Thr Pro Thr Asp Val Ser 115
120 125 Arg Asn Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe
Leu 130 135 140 Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly
Val Thr Val 145 150 155 160 Arg Asp Ser Leu Lys Lys Ala Leu Met Met
Arg Gly Leu Ile Pro Glu 165 170 175 Cys Cys Ala Val Tyr Arg Ile Gln
Asp Gly Glu Lys Lys Pro Ile Gly 180 185 190 Trp Asp Thr Asp Ile Ser
Trp Leu Thr Gly Glu Glu Leu His Val Glu 195 200 205 Val Leu Glu Asn
Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr 210 215 220 Phe Phe
Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln 225 230 235
240 Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser
245 250 255 Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp
Leu Leu 260 265 270 Phe Val Ser Lys Phe Phe Glu His His Pro Val Pro
Gln Glu Glu Ala 275 280 285 Phe Ser Ala Glu Thr Thr Leu Pro Ser Gly
Cys Ser Ser Ala Pro Pro 290 295 300 Ser Asp Ser Ile Gly Pro Gln Ile
Leu Thr Ser Pro Ser Pro Ser Lys 305 310 315 320 Ser Ile Pro Ile Pro
Gln Pro Phe Arg Pro Ala Asp Glu Asp His Arg 325 330 335 Asn Gln Phe
Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val His 340 345 350 Ile
Asn Thr Ile Glu Pro Val Asn Ile Asp Glu Lys Phe Pro Glu Val 355 360
365 Glu Leu Gln Asp Gln Arg Asp Leu Ile Arg Asp Gln Gly Phe Arg Gly
370 375 380 Asp Gly Ala Pro Leu Asn Gln Leu Met Arg Cys Leu Arg Lys
Tyr Gln 385 390 395 400 Ser Arg Thr Pro Ser Pro Leu Leu His Ser Val
Pro Ser Glu Ile Val 405 410 415 Phe Asp Phe Glu Pro Gly Pro Val Phe
Arg Gly Ser Thr Thr Gly Leu 420 425 430 Ser Ala Thr Pro Pro Ala Ser
Leu Pro Gly Ser Leu Thr Asn Val Lys 435 440 445 Ala Leu Gln Lys Ser
Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser 450 455 460
Ser Ser Ser Ser Thr Glu Asp Arg Ser Arg Met Lys Thr Leu Gly Arg 465
470 475 480 Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro Asp Gly Gln Ile
Thr Val 485 490 495 Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly Thr Val
Tyr Lys Gly Lys 500 505 510 Trp His Gly Asp Val Ala Val Lys Met Leu
Asn Val Thr Ala Pro Thr 515 520 525 Pro Gln Gln Leu Gln Ala Phe Lys
Asn Glu Val Gly Val Leu Arg Lys 530 535 540 Thr Arg His Val Asn Ile
Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro 545 550 555 560 Gln Leu Ala
Ile Val Thr Gln Trp Cys Glu Gly Ser Ser Leu Tyr His 565 570 575 His
Leu His Ile Ile Glu Thr Lys Phe Glu Met Ile Lys Leu Ile Asp 580 585
590 Ile Ala Arg Gln Thr Ala Gln Gly Met Asp Tyr Leu His Ala Lys Ser
595 600 605 Ile Ile His Arg Asp Leu Lys Ser Asn Asn Ile Phe Leu His
Glu Asp 610 615 620 Leu Thr Val Lys Ile Gly Asp Phe Gly Leu Ala Thr
Val Lys Ser Arg 625 630 635 640 Trp Ser Gly Ser His Gln Phe Glu Gln
Leu Ser Gly Ser Ile Leu Trp 645 650 655 Met Ala Pro Glu Val Ile Arg
Met Gln Asp Lys Asn Pro Tyr Ser Phe 660 665 670 Gln Ser Asp Val Tyr
Ala Phe Gly Ile Val Leu Tyr Glu Leu Met Thr 675 680 685 Gly Gln Leu
Pro Tyr Ser Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe 690 695 700 Met
Val Gly Arg Gly Tyr Leu Ser Pro Asp Leu Ser Lys Val Arg Ser 705 710
715 720 Asn Cys Pro Lys Ala Met Lys Arg Leu Met Ala Glu Cys Leu Lys
Lys 725 730 735 Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln Ile Leu Ala
Ser Ile Glu 740 745 750 Leu Leu Ala Arg Ser Leu Pro Lys Ile His Arg
Ser Ala Ser Glu Pro 755 760 765 Ser Leu Asn Arg Ala Gly Phe Gln Thr
Glu Asp Phe Ser Leu Tyr Ala 770 775 780 Cys Ala Ser Pro Lys Thr Pro
Ile Gln Ala Gly Gly Tyr Gly Glu Phe 785 790 795 800 Ala Ala Phe Lys
59728DNAMus musculus 5ccctcaggct cggctgcgcc ggggccgccg gcgggttcca
gaggtggcct ccgccccggc 60cgctccgccc acgccccccg cgcctccgcg cccgcctccg
cccgccctgc gcctcccttc 120cccctccccg ccccgcggcg gccgctcggc
ccggctcgcg cttcgaagat ggcggcgctg 180agtggcggcg gtggcagcag
cagcggtggc ggcggcggcg gtggcggcgg cggtggcggt 240ggcgacggcg
gcggcggcgc cgagcagggc caggctctgt tcaatggcga catggagccg
300gaggccggcg ctggcgccgc ggcctcttcg gctgcggacc cggccattcc
tgaagaggta 360tggaatatca agcaaatgat taagttgaca caggaacata
tagaggccct attggacaaa 420tttggtggag agcataaccc accatcaata
tacctggagg cctatgaaga gtacaccagc 480aagctagatg cccttcagca
aagagaacag cagcttttgg aatccctggt ttttcaaact 540cccacagatg
catcacggaa caaccccaag tcaccacaga aacctatcgt tagagtcttc
600ctgcccaaca aacagaggac agtggtaccc gcaagatgtg gtgttacagt
tcgagacagt 660ctaaagaaag cactgatgat gagaggtctc atcccagaat
gctgtgctgt ttacagaatt 720caggatggag agaagaaacc aattggctgg
gacacggaca tttcctggct tactggagag 780gagttacatg ttgaagtact
ggagaatgtc ccacttacaa cacacaactt tgtacggaaa 840acttttttca
ccttagcatt ttgtgacttt tgccgaaagc tgcttttcca gggtttccgt
900tgtcaaacat gtggttataa atttcaccag cgttgtagta cagaggttcc
actgatgtgt 960gtaaattatg accaacttga tttgctgttt gtctccaagt
tctttgagca tcacccagta 1020ccacaggagg aggcctcctt cccagagact
gcccttccat ctggatcctc ttccgcaccc 1080ccctcagact ctactgggcc
ccaaatcctc accagtccat ctccttcaaa atccattcca 1140attccacagc
ccttccgacc agcagatgaa gatcatcgca atcagtttgg gcaacgagac
1200cggtcctcct cagctcccaa tgttcatata aacacaattg agcctgtgaa
tatcgatgaa 1260aaattcccag aagtggaatt acaggatcaa agggatttga
ttagagacca ggggtttcgt 1320ggtgatggag cccccttgaa ccaactgatg
cgctgtcttc ggaaatacca atcccggact 1380cccagccccc tcctccattc
tgtccccagt gaaatagtgt ttgattttga gcctggccca 1440gtgttcagag
ggtcaaccac aggcttgtcc gccaccccgc ctgcctcatt acctggctca
1500ctcactaacg tgaaagcctt acagaaatct ccaggtcctc agcgggaaag
gaagtcatct 1560tcttcctcat cctcggagga cagaagtcgg atgaaaacac
ttggtagaag agattcaagt 1620gatgactggg agattcctga tggacagatt
acagtgggac agagaattgg atctgggtca 1680tttggaactg tctacaaggg
aaagtggcat ggtgatgtgg cagtgaaaat gttgaatgtg 1740acagcaccca
cacctcaaca gctacaggcc ttcaaaaatg aagtaggagt gctcaggaaa
1800actcgacatg tgaatatcct ccttttcatg ggctattcta caaagccaca
actggcaatt 1860gttacacagt ggtgtgaggg ctccagctta tatcaccatc
tccacatcat tgagaccaaa 1920tttgagatga tcaaacttat agatattgct
cggcagactg cacagggcat ggattactta 1980cacgccaagt caatcatcca
cagagacctc aagagtaata atatatttct tcatgaagac 2040ctcacggtaa
aaataggtga ctttggtcta gccacagtga aatctcggtg gagtgggtcc
2100catcagtttg aacagttgtc tggatctatt ttgtggatgg caccagaagt
aatcagaatg 2160caagataaaa acccgtatag ctttcagtca gacgtgtatg
cgtttgggat tgttctgtac 2220gaactgatga ccggccagct accttattca
aacatcaaca acagggatca gataattttt 2280atggtgggac gaggatacct
atctccagat ctcagtaagg tacggagtaa ctgtccaaaa 2340gccatgaaga
gattaatggc agagtgcctc aaaaagaaaa gagacgagag accactcttt
2400ccccaaattc tcgcctccat tgagctgctg gcccgctcat tgccaaaaat
tcaccgcagt 2460gcatcagaac cttccttgaa tcgggctggt ttccaaacag
aagattttag tctgtatgct 2520tgtgcttctc cgaaaacacc catccaagca
gggggatatg gagaatttgc agccttcaag 2580tagccagtcc atcatggcag
catctactct ttatttctta agtcttgtgt tcatacagtt 2640tgttaacatc
aaaacacagt tctgttcctc aaaaaatttt ttaaagatac aaaattttca
2700atgcataagt tcatgtggaa cagaatggaa tttcctattc aacaaaagag
ggaagaatgt 2760tttaggaacc agaattctct gctgcccgtg tttcttcttc
aacataacta tcacgtgcat 2820acaagtctgc ccattcccaa gaagaaagag
gagagaccct gaattctgcc cttttggtgg 2880tcaggcatga tggaaagaat
ttgctgctgc agcttgggaa aattgctatg gaaagtctgc 2940cagtcgactt
tgcccttcta accaccagat cagcctgtgg ctggtcatct gatggggcga
3000tttccatcac caagcatcgt tcttgcctat tctgggatta tgttgtggag
cactttccct 3060gtccagcacc gttcatttct gagggatgga gtaaatgcag
cattcccttg tgtagcgcct 3120gttcagtcct cagcagctgc tgtcacagcg
aagcttttta cagttaagtg gtgggggaga 3180gttgaggaga gcctgcctcg
gggcagagaa aagggggtgc tgcatcttct tcctcacctc 3240cagctctctc
acctcgggtt gccttgctca ctgggctccg cctaaccact caggctgctc
3300agtgctggca cacattgcct tcttttctca ttgggtccag caattgagga
gagggttggg 3360ggattgtttc ctcctcaatg tagcaaattc tcaggaaaat
acagtccata tcttcctctc 3420agctcttcca gtcaccaaat acttacgtgg
ctcctttgtc caggacataa aacaccgtgg 3480acaacaccta attaaaagcc
tacaaaactg cttactgaca gttttgaatg tgagacactt 3540gtgtaattta
aatgtaaggt acaggtttta atttctgagt ttcttctatt tttatttaaa
3600agaagaaaat aattttcagt tttaattgga ataaatgagt acttcccaca
agactatata 3660ccctgaaaat tatatttttg ttaattgtaa acaactttta
aagaataatt attatccttt 3720tctctaccta aaaattatgg ggaatcttag
cataatgaca attatttata ctttttaaat 3780aaatggtact tgctggatcc
acactaacat ctttgctaac aatcccattg tttcttccaa 3840cttaactcct
acactacatc ctacatcctc tttctagtct tttatctata atatgcaacc
3900taaaataaac gtggtggcgt ctccattcat tctccctctt cctgttttcc
ccaagcctgg 3960tcttcaaaag gttgggtaat cggtccctga gctccctagc
tggcaatgca actattaggg 4020acattggagt tgcaggagag caggaagcct
gtccccagct gttcttctag aaccctaaat 4080cttatctttg cacagatcaa
aagtatcacc tcgtcacagt tctccttagc ctttacttac 4140aggtaatata
aataaaaatc accatagtag taaagaaaac aactggatgg attgatgacc
4200agtacctctc agagccagga atcttgaatc tccaggattt atacgtgcaa
atttaaggag 4260atgtacttag caacttcaag ccaagaactt ccaaaatact
agcgaatcta aaataaaatg 4320gaattttgag ttatttttaa agttcaaatt
ataattgata ccactatgta tttaagccta 4380ctcacagcaa gttagatgga
ttttgctaaa ctcattgcca gactgtggtg gtggtggtgg 4440tagtgtgcac
ctttaatcca agcaactcag caatcagaat gaggtaaatc tctgtgaata
4500caaggcctgc ctagtctgca gcgctagttc caggatagcc agggctacac
acacaaaaac 4560cctctctcaa aaaaaacaaa attaattagt tgataataaa
aaataactaa agtatcatca 4620aaggaaggcc tactggaagt tttatatatt
cccagtaaat tgaaaaatat tctgaagtta 4680ttaaccagtt agcaacaatg
tgtttttaag tcttacataa acagagcaaa gtcttcaaat 4740gtttcagagc
tgagaagata attgtgcttg atatgaaaaa tagcctctcc atatgatgtg
4800ccacattgaa aggcgtcatt acccttttaa atacttctta atgtggcttt
gttcccttta 4860cccaggatta gctagaaaga gctaggtagg cttcggccac
agttgcacat ttcgggcctg 4920ctgaagaatg ggagctttga aggctggcct
tggtggagga gcccctcagt gctggagggt 4980ggggcgtgta cgcagcatgg
aagtggtcta gacagagtgc aaagggacag acttctttct 5040cattttagta
tagggtgatg tctcacttga aatgagaaag tagagttgat attaaacgaa
5100gctgtgccca gaaaccaggc tcagggtatt gtgagatttt ctttttaaat
agagaatata 5160aaagatagaa ataaatattt aaaccttcct tcttattttc
tatcaaatag atttttttta 5220tcatttgcaa acaacataaa aaaaggtttc
ttttgtgggg ttttctttcc ttcttttttt 5280tttttttttt tttttaagac
tgcagataat cttgttgagc tcctcggaaa atacaaggaa 5340gtccgtgttt
gtgcagagcg ctttatgagt aactgtatag acagtgtggc tgcttcactc
5400atcccagagg gctgcagctg tcggcccatg aagtggctgc agtgcctcgt
gagatctgct 5460ttgttttgtt tggagtgaag tctttgaaag gtttgagtgc
aactatatag gactgttttt 5520aaataagtag tattcctcat gaactttctc
attgttaagc tacaggaccc aaactctacc 5580actaagatat tattaacctc
aaaatgtagt ttatagaagg aatttgcaaa tagaatatcc 5640agttcgtact
tatatgcatc ttcaacaaag attctctgtg acttgttgga tttggttcct
5700gaacagccca tttctgtatt tgaggttagg agggcataat gaggcatcct
aaaagacaat 5760ctgatataaa ctgtatgcta gatgtatgct ggtaggggag
aaagcattct gtaaagacat 5820gatttaagac ttcagctctg tcaaccagaa
accttgtaaa tacttcctgt cttggtgcag 5880ccccgcccct ttgatcacac
gatgttgtct tgtgcttgtc agacactgtc agagctgctg 5940ttcgtccctc
tgcagatctc acctgtcccc actgcacacc cacctcctgc ctcttgcaga
6000cctcagcatc tagctttagt tggaaacagt tcagggttca ggtgacttct
taaaaaaaaa 6060aaaaaaccct acctcctcag aatgaggtaa tgaatagtta
tttatttaaa gtatgaagag 6120tcaggagcgc tcgaacatga aggtgattta
agatggttcc tttcgtgtgt attgtagctg 6180agcacttgtt tttgtcctaa
agggcattat acatttaagc agtgattctg tttaaagatg 6240tttttcttta
aaggtgtagc tcagagtatc tgttgttgga attggtgcca gagtctgctt
6300aatagatttc agaatcctaa gcttaagtca gtcgcatgaa gttaagtagt
tatggtaaca 6360ctttgctagc catgatataa ttctactttt taggagtagg
tttggcaaaa ctgtatgcct 6420tcaaagtgag ttggccacag ctttgtcaca
tgcacagata ctcatctgaa gagactgccc 6480agctaagagg gcggaaggat
accctttttt cctacgattc gcttctttgt ccacgttggc 6540attgttagta
ctagtttatc agcaccttga ccagcagatg tcaaccaata agctattttt
6600aaaaccatag ccagagatgg agaggtcact gtgagtagaa acagcaggac
gcttacagga 6660gtgaaatggt gtagggaggc tctagaaaaa tatcttgaca
atttgccaaa tgatcttact 6720gtgccttcat gatgcaataa aaaagctaac
attttagcag aaatcagtga tttacgaaga 6780gagtggccag tctggtttaa
ctcagctggg ataatatttt tagagtgcaa tttagactgc 6840gaagataaat
gcactaaaga gtttatagcc aattcacatt tgaaaaataa gaaaatggta
6900aattttcagt gaaatatttt tttaaagcac ataatcccta gtgtagccag
aaatatttac 6960cacatagagc agctaggctg agatacagtc cagtgacatt
tctagagaaa ccttttctac 7020tcccacgggc tcctcaaagc atggaaattt
tatacaaaat gtttgacatt ttaagatact 7080gctgtagttt agttttgaaa
tagtatgtgc tgagcagcaa tcatgtacta actcagagag 7140agaaaacaac
aacaaattgt gcatctgatt tgttttcaga gaaatgctgc caacttagat
7200actgagttct cagagcttca agtgtaaact tgcctcccaa gtcctgtttg
caaatgaagt 7260tggctagtgc tactgactgc tccagcacat gatggaaggc
agggggctgt ctctgaagtg 7320tcttctataa agggacaata gaatagtgag
agacctggtc agtgtgtgtc agctggacac 7380tccatgctat gggacttgca
tcttctgtcc tcaccatccc caagacattg tgctttcctc 7440agttgtcctc
tagctgtttc actcagacac caagatgaat tactgatgcc agaaggggcc
7500aaaatggcca gtgtgttttg ggggttgtat cagttgactg gacaataact
ttaatagttt 7560cagatcattt atttttactt ccattttgac agacatttaa
atggaaattt agtcctaact 7620tttgtcattt gaaaggaaaa attaacagtt
cctataagat acttttgagg tggaatctga 7680catcctaatt ttttttcttt
tcagtgggtt tgcagcgagg gtcttgtatg cactaggcaa 7740gggttctacc
actaagccac atttcccagg aaataaaatg ttaacagtta aaacatacac
7800acaaatacac aaacacctta ttaccacttt agtaaagtga gagatgtgcg
tcctttgtct 7860cagtctccac gatttcagct gccccttgta tgaataactc
agtctcgcta aactgtttac 7920ttttatttac ctggtttgac tagttgcagc
tatataacca gttgtgcatg aggacaacag 7980ccagtgtgtt tgttttgttt
ttggtttttt gtggtacatt ttttgtaaag aattctgtag 8040attgaagtgc
tctttgaaaa cagaactgag atatatttat tcttgttagc atcaaaaaac
8100attttgtgca aatgatttgc ttttcctggc aggctgagta ccatatccag
cgcccacaat 8160tgcgggttcc catctaccat gtccacaggg gagacagacg
ggaagcacat gaggggtgtg 8220tttacagagt tgtaggagtt atgtagttct
cttgttgcct tggaaatcac tgttgtttta 8280agactgttga acccgtgtgt
ttggctgggc tgtgagttac atgaagaaac tgcaaactag 8340catatgcaga
caaagctcac agactaggcg taaatggagg aaaatggacc aaaataaggc
8400agggtgacac ataaaccttg ggcttcggag aaaactaagg gtggagatga
actataatca 8460cctgaataca atgtaagagt gcaataagtg tgcttattct
aagctgtgaa cttcttttaa 8520atcattcctt tctaatacat ttatgtatgt
tccattgctg actaaaacca gctatgagaa 8580catatgcctt tttattcatg
ttaactacca gtttaagtgg ctaaccttaa tgtcttattt 8640atcttcattt
tgtattagtt tacataccag gtatgtgtgt gtgctgtact cttcttccct
8700ttatttgaaa acacttttca ctgggtcatc tccttggcca ttccacaaca
caactttggt 8760ttggctttca atgtcacctt atttgatggc ctgtgtccca
gtagcagaat ttatggtatt 8820cccattgctg gctgctcttc cgaccctttg
cttctacagc acttgtctct cctaagatag 8880tcagaaacta actgatcagg
ggatggactt caccattcat cgtgtctctt caattctatt 8940aaatagacca
ctcttgggct ttagaccagg aaaaaggaga cagctctagc catctaccaa
9000gcctcaccct aaaaggtcac ccgtacttct tggtctgagg acaagtctcc
actccagtaa 9060gggagagggg aggaaatgct tcctgtttga aatgcagtga
attcctatgg ctcctgtttc 9120accacccgca cctatggcaa cccatataca
ttcctcttgt ctgtaactgc caaaggttgg 9180gtttatgtca cttcagttcc
actcaagcat tgaaaaggtt ctcatggagt ctggggtgtg 9240cccagtgaaa
agatggggac tttttcatta tccacagacc tctctatacc tgctttgcaa
9300aaattataat ggagtaacta tttttaaagc ttatttttca attcataaga
aaaagacatt 9360tattttcaat caaatggatg atgtctctta tcccttatcc
ctcaatgttt gcttgaattt 9420tgtttgttcc ctatacctac tccctaattc
tttagttcct tcctgctcag gtcccttcat 9480ttgtactttg gagtttttct
catgtaaatt tgtataatgg aaaatattgt tcagtttgga 9540tagaaagcat
ggagaaataa ataaaaaaag atagctgaaa atcaaattga agaaatttat
9600ttctgtgtaa agttatttaa aaactctgta ttatatttaa agaaaaaagc
ccaacccccc 9660aaaaagtgct atgtaattga tgtgaatatg cgaatactgc
tataataaag attgactgca 9720tggagaaa 97286804PRTMus musculus 6Met Ala
Ala Leu Ser Gly Gly Gly Gly Ser Ser Ser Gly Gly Gly Gly 1 5 10 15
Gly Gly Gly Gly Gly Gly Gly Gly Gly Asp Gly Gly Gly Gly Ala Glu 20
25 30 Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly
Ala 35 40 45 Gly Ala Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro
Glu Glu Val 50 55 60 Trp Asn Ile Lys Gln Met Ile Lys Leu Thr Gln
Glu His Ile Glu Ala 65 70 75 80 Leu Leu Asp Lys Phe Gly Gly Glu His
Asn Pro Pro Ser Ile Tyr Leu 85 90 95 Glu Ala Tyr Glu Glu Tyr Thr
Ser Lys Leu Asp Ala Leu Gln Gln Arg 100 105 110 Glu Gln Gln Leu Leu
Glu Ser Leu Val Phe Gln Thr Pro Thr Asp Ala 115 120 125 Ser Arg Asn
Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe 130 135 140 Leu
Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr 145 150
155 160 Val Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile
Pro 165 170 175 Glu Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys
Lys Pro Ile 180 185 190 Gly Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly
Glu Glu Leu His Val 195 200 205 Glu Val Leu Glu Asn Val Pro Leu Thr
Thr His Asn Phe Val Arg Lys 210 215 220 Thr Phe Phe Thr Leu Ala Phe
Cys Asp Phe Cys Arg Lys Leu Leu Phe 225 230 235 240 Gln Gly Phe Arg
Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys 245 250 255 Ser Thr
Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu 260 265 270
Leu Phe Val Ser Lys Phe Phe Glu His His Pro Val Pro Gln Glu Glu 275
280 285 Ala Ser Phe Pro Glu Thr Ala Leu Pro Ser Gly Ser Ser Ser Ala
Pro 290 295 300 Pro Ser Asp Ser Thr Gly Pro Gln Ile Leu Thr Ser Pro
Ser Pro Ser 305 310 315 320 Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg
Pro Ala Asp Glu Asp His 325 330 335 Arg Asn Gln Phe Gly Gln Arg Asp
Arg Ser Ser Ser Ala Pro Asn Val 340 345 350 His Ile Asn Thr Ile Glu
Pro Val Asn Ile Asp Glu Lys Phe Pro Glu 355 360 365 Val Glu Leu Gln
Asp Gln Arg Asp Leu Ile Arg Asp Gln Gly Phe Arg 370 375 380 Gly Asp
Gly Ala Pro Leu Asn Gln Leu Met Arg Cys Leu Arg Lys Tyr 385 390 395
400 Gln Ser Arg Thr Pro Ser Pro Leu Leu His Ser Val Pro Ser Glu Ile
405 410 415 Val Phe Asp Phe Glu Pro Gly Pro Val Phe Arg Gly Ser Thr
Thr Gly 420 425 430 Leu Ser Ala Thr Pro Pro Ala Ser Leu Pro Gly Ser
Leu Thr Asn Val 435 440 445 Lys Ala Leu Gln Lys Ser Pro Gly Pro Gln
Arg Glu Arg Lys Ser Ser 450 455 460
Ser Ser Ser Ser Ser Glu Asp Arg Ser Arg Met Lys Thr Leu Gly Arg 465
470 475 480 Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro Asp Gly Gln Ile
Thr Val 485 490 495 Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly Thr Val
Tyr Lys Gly Lys 500 505 510 Trp His Gly Asp Val Ala Val Lys Met Leu
Asn Val Thr Ala Pro Thr 515 520 525 Pro Gln Gln Leu Gln Ala Phe Lys
Asn Glu Val Gly Val Leu Arg Lys 530 535 540 Thr Arg His Val Asn Ile
Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro 545 550 555 560 Gln Leu Ala
Ile Val Thr Gln Trp Cys Glu Gly Ser Ser Leu Tyr His 565 570 575 His
Leu His Ile Ile Glu Thr Lys Phe Glu Met Ile Lys Leu Ile Asp 580 585
590 Ile Ala Arg Gln Thr Ala Gln Gly Met Asp Tyr Leu His Ala Lys Ser
595 600 605 Ile Ile His Arg Asp Leu Lys Ser Asn Asn Ile Phe Leu His
Glu Asp 610 615 620 Leu Thr Val Lys Ile Gly Asp Phe Gly Leu Ala Thr
Val Lys Ser Arg 625 630 635 640 Trp Ser Gly Ser His Gln Phe Glu Gln
Leu Ser Gly Ser Ile Leu Trp 645 650 655 Met Ala Pro Glu Val Ile Arg
Met Gln Asp Lys Asn Pro Tyr Ser Phe 660 665 670 Gln Ser Asp Val Tyr
Ala Phe Gly Ile Val Leu Tyr Glu Leu Met Thr 675 680 685 Gly Gln Leu
Pro Tyr Ser Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe 690 695 700 Met
Val Gly Arg Gly Tyr Leu Ser Pro Asp Leu Ser Lys Val Arg Ser 705 710
715 720 Asn Cys Pro Lys Ala Met Lys Arg Leu Met Ala Glu Cys Leu Lys
Lys 725 730 735 Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln Ile Leu Ala
Ser Ile Glu 740 745 750 Leu Leu Ala Arg Ser Leu Pro Lys Ile His Arg
Ser Ala Ser Glu Pro 755 760 765 Ser Leu Asn Arg Ala Gly Phe Gln Thr
Glu Asp Phe Ser Leu Tyr Ala 770 775 780 Cys Ala Ser Pro Lys Thr Pro
Ile Gln Ala Gly Gly Tyr Gly Glu Phe 785 790 795 800 Ala Ala Phe Lys
72232DNAOryctolagus cuniculus 7atggggaatg tgtggaatat caaacaaatg
attaagttga cacaggagca tatagaggcc 60ctattggaca aatttggtgg ggagcataat
ccaccatcaa tatatctgga ggcctacgaa 120gaatacacca gcaagctaga
tgccctccaa caaagagaac agcagttatt ggaatcccta 180gtttttcaaa
atcccacaga tgtgtcacgg agcaacccca agtcaccaca aaaacctatt
240gttagagtct tcctgcccaa caaacagagg acagtggtac ctgcaagatg
tggagttacg 300gttcgagaca gtctaaagaa agcgctgatg atgagaggtc
tgatcccaga atgctgtgct 360gtttacagaa ttcaggatgg agagaagaag
ccaattggct gggacactga tatttcctgg 420ctcactggag aagagctgca
tgtggaagtg ttagagaatg tcccactcac cacacataac 480tttgtacgga
aaactttttt caccttagca ttttgtgact tctgtagaaa gctgcttttc
540cagggtttcc gctgtcaaac atgtggctac aaatttcacc agcgttgtag
tacggaagtt 600ccactgatgt gtgttaatta tgaccaactt gatttgctgt
ttgtctccaa gttctttgaa 660caccacccag taccacagga ggaggcctcc
ttagcagaga ctgccctcac atctgggtca 720tcgccttccg cacctccctc
agactctatt gggcaccaaa ttctcaccag tccgtcccct 780tcaaaatcca
ttccgattcc acagtccttc cgaccagcag atgaagatca tcgaaatcag
840tttgggcaac gagaccggtc ttcatcagcg cctaatgttc acattaacac
aatagaacct 900gtcaatattg atgaaaaatt cccagaagtg gaattacagg
atcaaaggga cttgattaga 960gaccaagggt ttcgtggtga tggagcccct
ttgaaccagc tgatgcgctg tcttcggaaa 1020taccaatccc ggactcccag
tcccctccta ccttctgtcc ccagtgacat agtgtttgat 1080tttgagcctg
gcccagtgtt cagaggatcg accacgggtt tgtctgccac tccccctgcc
1140tcattacctg gctcactcac tagtgtgaaa gctgtacaga gatccccagg
acctcagcga 1200gagaggaagt cgtcttcctc ctcagaagac aggaatcgaa
tgaaaactct tggtagacgg 1260gattcaagtg atgattggga gattcctgat
gggcagatca ccgtgggaca gagaattgga 1320tctggatcat ttggaaccgt
ctacaaggga aaatggcacg gtgatgtggc agtaaaaatg 1380ttgaatgtga
cagcacctac acctcagcag ttacaggcct tcaaaaatga agtaggagta
1440ctcaggaaaa cacgacatgt gaatatccta cttttcatgg gctattccac
aaagccacag 1500ctggctattg ttacccagtg gtgtgagggc tccagtttat
atcaccatct ccacatcatt 1560gagaccaaat tcgagatgat caaacttata
gatattgcac ggcagactgc acagggcatg 1620gattacttac acgccaagtc
aatcatccac agagacctca agagtaataa tatatttctt 1680catgaagacc
tcacagtaaa aataggtgat tttggtctag ccacagtgaa atctcgatgg
1740agtgggtccc atcagtttga acaattgtct ggatccattt tgtggatggc
accagaagta 1800atcagaatgc aagacaaaaa cccatatagc tttcagtcag
atgtatatgc atttgggatt 1860gttctgtatg aattgatgac tgggcagtta
ccttactcaa acatcaacaa cagggaccag 1920atcattttta tggtgggacg
tggctacctg tctccagacc tcagtaaggt acggagtaac 1980tgtccgaaag
ccatgaagag attaatggca gagtgcctca aaaagaaaag agatgagaga
2040ccactctttc cccaaattct cgcctccatt gagctgctgg cccgctcatt
gccaaaaatc 2100caccgcagtg catcagaacc ctccttgaat cgggctggtt
tccagacaga ggattttagt 2160ctatatgctt gtgcttctcc aaaaacaccc
atccaggcag ggggatatgg agaatttgca 2220gccttcaagt ag
22328743PRTOryctolagus cuniculus 8Met Gly Asn Val Trp Asn Ile Lys
Gln Met Ile Lys Leu Thr Gln Glu 1 5 10 15 His Ile Glu Ala Leu Leu
Asp Lys Phe Gly Gly Glu His Asn Pro Pro 20 25 30 Ser Ile Tyr Leu
Glu Ala Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala 35 40 45 Leu Gln
Gln Arg Glu Gln Gln Leu Leu Glu Ser Leu Val Phe Gln Asn 50 55 60
Pro Thr Asp Val Ser Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile 65
70 75 80 Val Arg Val Phe Leu Pro Asn Lys Gln Arg Thr Val Val Pro
Ala Arg 85 90 95 Cys Gly Val Thr Val Arg Asp Ser Leu Lys Lys Ala
Leu Met Met Arg 100 105 110 Gly Leu Ile Pro Glu Cys Cys Ala Val Tyr
Arg Ile Gln Asp Gly Glu 115 120 125 Lys Lys Pro Ile Gly Trp Asp Thr
Asp Ile Ser Trp Leu Thr Gly Glu 130 135 140 Glu Leu His Val Glu Val
Leu Glu Asn Val Pro Leu Thr Thr His Asn 145 150 155 160 Phe Val Arg
Lys Thr Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg 165 170 175 Lys
Leu Leu Phe Gln Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe 180 185
190 His Gln Arg Cys Ser Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp
195 200 205 Gln Leu Asp Leu Leu Phe Val Ser Lys Phe Phe Glu His His
Pro Val 210 215 220 Pro Gln Glu Glu Ala Ser Leu Ala Glu Thr Ala Leu
Thr Ser Gly Ser 225 230 235 240 Ser Pro Ser Ala Pro Pro Ser Asp Ser
Ile Gly His Gln Ile Leu Thr 245 250 255 Ser Pro Ser Pro Ser Lys Ser
Ile Pro Ile Pro Gln Ser Phe Arg Pro 260 265 270 Ala Asp Glu Asp His
Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser 275 280 285 Ser Ala Pro
Asn Val His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp 290 295 300 Glu
Lys Phe Pro Glu Val Glu Leu Gln Asp Gln Arg Asp Leu Ile Arg 305 310
315 320 Asp Gln Gly Phe Arg Gly Asp Gly Ala Pro Leu Asn Gln Leu Met
Arg 325 330 335 Cys Leu Arg Lys Tyr Gln Ser Arg Thr Pro Ser Pro Leu
Leu Pro Ser 340 345 350 Val Pro Ser Asp Ile Val Phe Asp Phe Glu Pro
Gly Pro Val Phe Arg 355 360 365 Gly Ser Thr Thr Gly Leu Ser Ala Thr
Pro Pro Ala Ser Leu Pro Gly 370 375 380 Ser Leu Thr Ser Val Lys Ala
Val Gln Arg Ser Pro Gly Pro Gln Arg 385 390 395 400 Glu Arg Lys Ser
Ser Ser Ser Ser Glu Asp Arg Asn Arg Met Lys Thr 405 410 415 Leu Gly
Arg Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro Asp Gly Gln 420 425 430
Ile Thr Val Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly Thr Val Tyr 435
440 445 Lys Gly Lys Trp His Gly Asp Val Ala Val Lys Met Leu Asn Val
Thr 450 455 460 Ala Pro Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu
Val Gly Val 465 470 475 480 Leu Arg Lys Thr Arg His Val Asn Ile Leu
Leu Phe Met Gly Tyr Ser 485 490 495 Thr Lys Pro Gln Leu Ala Ile Val
Thr Gln Trp Cys Glu Gly Ser Ser 500 505 510 Leu Tyr His His Leu His
Ile Ile Glu Thr Lys Phe Glu Met Ile Lys 515 520 525 Leu Ile Asp Ile
Ala Arg Gln Thr Ala Gln Gly Met Asp Tyr Leu His 530 535 540 Ala Lys
Ser Ile Ile His Arg Asp Leu Lys Ser Asn Asn Ile Phe Leu 545 550 555
560 His Glu Asp Leu Thr Val Lys Ile Gly Asp Phe Gly Leu Ala Thr Val
565 570 575 Lys Ser Arg Trp Ser Gly Ser His Gln Phe Glu Gln Leu Ser
Gly Ser 580 585 590 Ile Leu Trp Met Ala Pro Glu Val Ile Arg Met Gln
Asp Lys Asn Pro 595 600 605 Tyr Ser Phe Gln Ser Asp Val Tyr Ala Phe
Gly Ile Val Leu Tyr Glu 610 615 620 Leu Met Thr Gly Gln Leu Pro Tyr
Ser Asn Ile Asn Asn Arg Asp Gln 625 630 635 640 Ile Ile Phe Met Val
Gly Arg Gly Tyr Leu Ser Pro Asp Leu Ser Lys 645 650 655 Val Arg Ser
Asn Cys Pro Lys Ala Met Lys Arg Leu Met Ala Glu Cys 660 665 670 Leu
Lys Lys Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln Ile Leu Ala 675 680
685 Ser Ile Glu Leu Leu Ala Arg Ser Leu Pro Lys Ile His Arg Ser Ala
690 695 700 Ser Glu Pro Ser Leu Asn Arg Ala Gly Phe Gln Thr Glu Asp
Phe Ser 705 710 715 720 Leu Tyr Ala Cys Ala Ser Pro Lys Thr Pro Ile
Gln Ala Gly Gly Tyr 725 730 735 Gly Glu Phe Ala Ala Phe Lys 740
92186DNACavia porcellus 9atggcggcgc tcagcggcgg cggtggcgcg
gagcagggcc aggctctgtt caacggggac 60atggagctcg aggccggcgc cggcgccgca
gcctcttcgg ctgcagaccc tgccattccc 120gaggaggtat ggaatatcaa
acaaatgatt aagttgacgc aggaacacat agaggcccta 180ttggacaaat
ttggtggaga gcataatcca ccatcaatat acctggaggc ctatgaagaa
240tacaccagca aactagatgc cctccaacaa agagaacagc agttactgga
atccctcggg 300aatggaactg atttttctgt ttctagctct gcatcactgg
acaccgttac atcttcttct 360tcttctagcc tttcagtact accttcatct
ctttcagttt ttcaaaatcc tacagatgtg 420tcacggagca accccaaatc
accacaaaaa cctattgtta gagtcttcct gcccaacaaa 480cagaggacag
tggtacctgc aaggtgtgga gttacagtcc gagacagtct gaagaaagca
540ctcatgatga gaggtcttat cccagagtgc tgtgctgtgt acagaattca
ggatggagaa 600aagaaaccaa ttggctggga cactgacatt tcctggctta
ctggggaaga attacatgta 660gaagtattgg agaatgttcc acttacaaca
cacaattttg tatgtatctt tatatttttt 720ttgctgtttg tctccaagtt
ctttgaacac cacccaatac cacaggagga ggcttcctta 780gcagagacca
cccttacatc tggatcatcc ccttctgcac ccccctcaga gtccattggg
840cccccaattc tcaccagccc atctccttca aaatccattc caattccaca
gcctttccgg 900ccaggagagg aagatcatcg aaatcaattt gggcagcgag
accggtcctc atctgctccc 960aatgtgcata taaacacaat agaacctgtc
aatattgatg atttgattag agaccaaggg 1020tttcgtagtg atggaggatc
aactacaggt ttgtctgcca ccccacctgc ctcattacct 1080ggctcactca
ctaatgtgaa agccttacag aaatctccag gacctcagcg agaaaggaag
1140tcatcttcat cctcagaaga cagaaatcga atgaaaacgc ttggtagacg
ggactcaagt 1200gatgattggg agattcctga tgggcagatt acagtgggac
aaagaattgg atctgggtca 1260tttggaacag tctacaaggg gaagtggcat
ggtgacgtgg cagtgaaaat gttgaatgtg 1320acagcaccca cacctcaaca
gttacaggcc ttcaaaaatg aagtaggagt actcaggaaa 1380acacgacatg
tgaatatcct actcttcatg ggctattcca caaagccaca gctagctatt
1440gttacccagt ggtgtgaggg ctccagctta taccaccatc tccacatcat
cgagaccaaa 1500tttgagatga tcaaacttat agatattgca cgacagactg
cccagggcat ggattactta 1560cacgccaagt caatcatcca cagagacctc
aagagtaata atatatttct tcacgaagac 1620ctcacggtta aaataggtga
ttttggtcta gccacagtga aatctcgatg gagtgggtcc 1680catcagtttg
aacagttgtc tggatccatt ttgtggatgg caccagaagt aatcagaatg
1740cgagataaaa acccatacag ttttcagtcc gatgtatatg catttgggat
tgttctatat 1800gaattgatga ctgggcagtt accctattca aatatcaaca
acagggacca gataattttt 1860atggtgggac gaggatatct atctccagat
ctcagcaagg tacggagtaa ctgtccaaaa 1920gccatgaaga ggttaatggc
ggagtgcctc aaaaagaaaa gagatgagag accactcttt 1980ccccaaattc
tcgcctctat tgagctgctg gcccgctcat tgccaaaaat tcaccgcagt
2040gcatcagaac cctccttgaa tcgggctggt ttccaaacag aggattttag
tctctatgct 2100tgtgcttctc caaaaacacc catccaggca gggggatatg
gtgcgtttcc tgtccactga 2160tgcaaattaa atgagtgaga aataaa
218610719PRTCavia porcellus 10Met Ala Ala Leu Ser Gly Gly Gly Gly
Ala Glu Gln Gly Gln Ala Leu 1 5 10 15 Phe Asn Gly Asp Met Glu Leu
Glu Ala Gly Ala Gly Ala Ala Ala Ser 20 25 30 Ser Ala Ala Asp Pro
Ala Ile Pro Glu Glu Val Trp Asn Ile Lys Gln 35 40 45 Met Ile Lys
Leu Thr Gln Glu His Ile Glu Ala Leu Leu Asp Lys Phe 50 55 60 Gly
Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala Tyr Glu Glu 65 70
75 80 Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln Gln Leu
Leu 85 90 95 Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser
Ser Ala Ser 100 105 110 Leu Asp Thr Val Thr Ser Ser Ser Ser Ser Ser
Leu Ser Val Leu Pro 115 120 125 Ser Ser Leu Ser Val Phe Gln Asn Pro
Thr Asp Val Ser Arg Ser Asn 130 135 140 Pro Lys Ser Pro Gln Lys Pro
Ile Val Arg Val Phe Leu Pro Asn Lys 145 150 155 160 Gln Arg Thr Val
Val Pro Ala Arg Cys Gly Val Thr Val Arg Asp Ser 165 170 175 Leu Lys
Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu Cys Cys Ala 180 185 190
Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly Trp Asp Thr 195
200 205 Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu Val Leu
Glu 210 215 220 Asn Val Pro Leu Thr Thr His Asn Phe Val Cys Ile Phe
Ile Phe Phe 225 230 235 240 Leu Leu Phe Val Ser Lys Phe Phe Glu His
His Pro Ile Pro Gln Glu 245 250 255 Glu Ala Ser Leu Ala Glu Thr Thr
Leu Thr Ser Gly Ser Ser Pro Ser 260 265 270 Ala Pro Pro Ser Glu Ser
Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser 275 280 285 Pro Ser Lys Ser
Ile Pro Ile Pro Gln Pro Phe Arg Pro Gly Glu Glu 290 295 300 Asp His
Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro 305 310 315
320 Asn Val His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile
325 330 335 Arg Asp Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly
Leu Ser 340 345 350 Ala Thr Pro Pro Ala Ser Leu Pro Gly Ser Leu Thr
Asn Val Lys Ala 355 360 365 Leu Gln Lys Ser Pro Gly Pro Gln Arg Glu
Arg Lys Ser Ser Ser Ser 370 375 380 Ser Glu Asp Arg Asn Arg Met Lys
Thr Leu Gly Arg Arg Asp Ser Ser 385 390 395 400 Asp Asp Trp Glu Ile
Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile 405 410 415 Gly Ser Gly
Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp 420 425 430 Val
Ala Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu 435 440
445 Gln Ala Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val
450 455 460 Asn Ile Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu
Ala Ile 465 470 475 480 Val Thr Gln Trp Cys Glu Gly Ser Ser Leu Tyr
His His Leu His Ile 485 490 495 Ile Glu Thr Lys Phe Glu Met Ile Lys
Leu Ile Asp Ile Ala Arg Gln 500 505 510 Thr Ala Gln Gly Met Asp Tyr
Leu His Ala Lys Ser Ile
Ile His Arg 515 520 525 Asp Leu Lys Ser Asn Asn Ile Phe Leu His Glu
Asp Leu Thr Val Lys 530 535 540 Ile Gly Asp Phe Gly Leu Ala Thr Val
Lys Ser Arg Trp Ser Gly Ser 545 550 555 560 His Gln Phe Glu Gln Leu
Ser Gly Ser Ile Leu Trp Met Ala Pro Glu 565 570 575 Val Ile Arg Met
Arg Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val 580 585 590 Tyr Ala
Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro 595 600 605
Tyr Ser Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe Met Val Gly Arg 610
615 620 Gly Tyr Leu Ser Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro
Lys 625 630 635 640 Ala Met Lys Arg Leu Met Ala Glu Cys Leu Lys Lys
Lys Arg Asp Glu 645 650 655 Arg Pro Leu Phe Pro Gln Ile Leu Ala Ser
Ile Glu Leu Leu Ala Arg 660 665 670 Ser Leu Pro Lys Ile His Arg Ser
Ala Ser Glu Pro Ser Leu Asn Arg 675 680 685 Ala Gly Phe Gln Thr Glu
Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro 690 695 700 Lys Thr Pro Ile
Gln Ala Gly Gly Tyr Gly Ala Phe Pro Val His 705 710 715
113229DNACanis familiaris 11gtaatgctgg attttcatgg aataagtttg
acctgtgctg cagtggcctc cagcaaggta 60cccgcaagat gtggagttac agtccgggac
agtctaaaga aagctctgat gatgagaggt 120ctaatcccag agtgctgtgc
tgtttacaga attcaggatg gagagaagaa accgattggc 180tgggacactg
atatttcctg gctcactgga gaggaattgc atgtagaagt gttggaaaat
240gttccgctta ccacacacaa ctttgtacgg aaaacttttt tcaccttagc
attttgtgac 300ttttgtcgaa agctgctttt ccagggtttt cgctgtcaaa
catgtggtta taaatttcac 360cagcgttgta gtacagaggt tccactgatg
tgtgttaatt atgaccaact tgatttgctg 420tttgtctcca agttctttga
acaccaccca ataccacagg aggaggcctc catagcagag 480actgccctta
cgtctggatc atccccttct gctcccccct ccgattctcc tgggccccca
540attctgacca gtccgtctcc ttcaaaatcc attccaattc cacagccttt
ccgaccagca 600gatgaagatc atcgaaatca gtttggacaa cgagaccggt
cctcatcagc tccaaatgtg 660catataaaca caatagaacc cgtcaacatt
gatgacttga ttagagacca agggtttcgt 720agtgatggag gatcaaccac
aggtttgtct gccacccccc ctgcctcatt gcctggctca 780ctcactaatg
taaaagcatt acagaaatct ccaggacctc agcgggaaag aaaatcatct
840tcatcctcag aagataggaa tcgaatgaaa acacttggta gacgggattc
aagtgatgat 900tgggagatac ctgatgggca gatcacagtg ggacagagaa
ttggatccgg gtcatttggg 960acagtctaca agggaaagtg gcatggtgac
gtggcagtga aaatgttgaa tgtgacagca 1020cccacacctc agcagttaca
ggccttcaaa aatgaagtag gagtactcag gaaaactcga 1080catgtgaata
tcctactctt tatgggctat tcaacaaagc cccaactggc tattgttacc
1140cagtggtgtg agggctccag cttatatcac catctccaca tcattgagac
caaatttgag 1200atgataaagc ttatagatat tgcacggcag actgcacagg
gcatggatta cttacacgcc 1260aagtcaatca tccacagaga cctcaagagt
aataatattt ttcttcatga agacctcaca 1320gtaaaaatag gtgattttgg
tctagccaca gtgaaatctc gatggagtgg gtcccatcag 1380tttgaacagt
tgtctggatc cattttgtgg atggcaccag aagtgatccg aatgcaagac
1440aaaaacccat atagcttcca gtcagatgta tacgcatttg ggattgttct
atatgaattg 1500atgacagggc agttacctta ttcaaacatc aacaacaggg
accagataat ttttatggtg 1560ggacgaggat atctttctcc agatctcagt
aaggtacgga gtaactgtcc aaaagccatg 1620aagagattga tggcagagtg
cctaaaaaag aaaagagatg agaggccact ctttccccaa 1680attctcgcct
ctattgagct gctggcccgc tcattgccaa aaattcaccg cagtgcatca
1740gaaccctcct tgaatcgggc tggcttccaa acagaggatt ttagtctcta
tgcttgcgct 1800tctccaaaaa cacccatcca ggcaggggga tacggagaat
ttgcagcctt caagtagcca 1860caccatcatg gcaacaacta ctcttatttc
ttaagtcttg tgttcgtaca atttgttaac 1920atcaaaacac agttctgttc
ctcaaatctt tttttaaaga tacagaattt tcaatgcata 1980agctggtgtg
gaacagaatg gaatttccca tccaacaaaa gagggaagaa tgttttagga
2040accagaattc tctgctgcca gtgtttcttc ttcaacacaa ataccacgtg
catacaagtc 2100tgcccactcc caggaaggaa gaggagagcc tgagttctga
ccttttgatg gtcaggcatg 2160atggaaagaa actgctgcta cagcttggga
gattggctgt ggagagcctg cccgtcagct 2220ctgcccttct aaccgccaga
tgagtgtgtg gctggtcacc tgacagggca gctgcaatcg 2280ccaagcatcg
ttctctttcc tgtcctggga ttttgtcgtg gagctctttc cccctagtca
2340ccaccggttc atttctgagg gatggaacaa aaatgcagca tggcctttct
gtgtggtgca 2400tgtccggtct ttgacaaatt tttatcaagt gaagctcttg
tatttaaatg gagaatgaga 2460ggcgaggggg ggggatcacg ttttggtgta
ggggcaaagg gaatgctgca tctttttcct 2520gacccactgg gtttctggcc
tttgtttcct tgctcactga gggtgtctgc ctataaccac 2580gcaggctgga
aagtgctggc acacattgcc ttctcttctc actgggtcca gcaatgaaga
2640caagtgttgg ggattttttt ttttgccctc cacaatgtag caagttctca
ggaaaataca 2700gttaatatct tcctcctaag ctcttccagt catcaagtac
ttatgtggct actttgtcca 2760gggcacaaaa tgccatggcg gtatccaatt
aaaagcctac aaaactgctt gataacagtt 2820ttgaatgtgt gagacattta
tgtaatttaa atgtaaggta caagttttaa tttctgagtt 2880tctctattat
atttttatta aaaagaaaat aattttcaga tttaattgaa ttggaataaa
2940ataatacttc ccaccagaat tatatatcct ggaaaattgt atttttgtta
tataaacaac 3000ttttaaagaa agatcattat ccttttctct acctaaatat
ggggagtctt agcataatga 3060cagatattta taatttttaa attaatggta
cttgctggat ccacactaac atctttgcta 3120atatctcatg ttttcctcca
acttactcct acactacatc ctccatcctc tttccagtct 3180tttatctaga
atatgcaacc taaaataaaa atggtggtgt ctccattca 322912617PRTCanis
familiaris 12Met Leu Asp Phe His Gly Ile Ser Leu Thr Cys Ala Ala
Val Ala Ser 1 5 10 15 Ser Lys Val Pro Ala Arg Cys Gly Val Thr Val
Arg Asp Ser Leu Lys 20 25 30 Lys Ala Leu Met Met Arg Gly Leu Ile
Pro Glu Cys Cys Ala Val Tyr 35 40 45 Arg Ile Gln Asp Gly Glu Lys
Lys Pro Ile Gly Trp Asp Thr Asp Ile 50 55 60 Ser Trp Leu Thr Gly
Glu Glu Leu His Val Glu Val Leu Glu Asn Val 65 70 75 80 Pro Leu Thr
Thr His Asn Phe Val Arg Lys Thr Phe Phe Thr Leu Ala 85 90 95 Phe
Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln Gly Phe Arg Cys Gln 100 105
110 Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser Thr Glu Val Pro Leu
115 120 125 Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu Phe Val Ser
Lys Phe 130 135 140 Phe Glu His His Pro Ile Pro Gln Glu Glu Ala Ser
Ile Ala Glu Thr 145 150 155 160 Ala Leu Thr Ser Gly Ser Ser Pro Ser
Ala Pro Pro Ser Asp Ser Pro 165 170 175 Gly Pro Pro Ile Leu Thr Ser
Pro Ser Pro Ser Lys Ser Ile Pro Ile 180 185 190 Pro Gln Pro Phe Arg
Pro Ala Asp Glu Asp His Arg Asn Gln Phe Gly 195 200 205 Gln Arg Asp
Arg Ser Ser Ser Ala Pro Asn Val His Ile Asn Thr Ile 210 215 220 Glu
Pro Val Asn Ile Asp Asp Leu Ile Arg Asp Gln Gly Phe Arg Ser 225 230
235 240 Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr Pro Pro Ala Ser
Leu 245 250 255 Pro Gly Ser Leu Thr Asn Val Lys Ala Leu Gln Lys Ser
Pro Gly Pro 260 265 270 Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu
Asp Arg Asn Arg Met 275 280 285 Lys Thr Leu Gly Arg Arg Asp Ser Ser
Asp Asp Trp Glu Ile Pro Asp 290 295 300 Gly Gln Ile Thr Val Gly Gln
Arg Ile Gly Ser Gly Ser Phe Gly Thr 305 310 315 320 Val Tyr Lys Gly
Lys Trp His Gly Asp Val Ala Val Lys Met Leu Asn 325 330 335 Val Thr
Ala Pro Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu Val 340 345 350
Gly Val Leu Arg Lys Thr Arg His Val Asn Ile Leu Leu Phe Met Gly 355
360 365 Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr Gln Trp Cys Glu
Gly 370 375 380 Ser Ser Leu Tyr His His Leu His Ile Ile Glu Thr Lys
Phe Glu Met 385 390 395 400 Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr
Ala Gln Gly Met Asp Tyr 405 410 415 Leu His Ala Lys Ser Ile Ile His
Arg Asp Leu Lys Ser Asn Asn Ile 420 425 430 Phe Leu His Glu Asp Leu
Thr Val Lys Ile Gly Asp Phe Gly Leu Ala 435 440 445 Thr Val Lys Ser
Arg Trp Ser Gly Ser His Gln Phe Glu Gln Leu Ser 450 455 460 Gly Ser
Ile Leu Trp Met Ala Pro Glu Val Ile Arg Met Gln Asp Lys 465 470 475
480 Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala Phe Gly Ile Val Leu
485 490 495 Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn
Asn Arg 500 505 510 Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr Leu
Ser Pro Asp Leu 515 520 525 Ser Lys Val Arg Ser Asn Cys Pro Lys Ala
Met Lys Arg Leu Met Ala 530 535 540 Glu Cys Leu Lys Lys Lys Arg Asp
Glu Arg Pro Leu Phe Pro Gln Ile 545 550 555 560 Leu Ala Ser Ile Glu
Leu Leu Ala Arg Ser Leu Pro Lys Ile His Arg 565 570 575 Ser Ala Ser
Glu Pro Ser Leu Asn Arg Ala Gly Phe Gln Thr Glu Asp 580 585 590 Phe
Ser Leu Tyr Ala Cys Ala Ser Pro Lys Thr Pro Ile Gln Ala Gly 595 600
605 Gly Tyr Gly Glu Phe Ala Ala Phe Lys 610 615 131889DNACanis
familiaris 13ggaatatcaa acaaatgatt aagttgacac aggaacatat agaagcccta
ttggacaagt 60ttggtgggga gcataatcca ccatcaatat atctggaggc ctatgaagaa
tacaccagca 120aactagatgc cctccaacag cgagaacaac agttattgga
atccctgggg aatggaactg 180atttttctgt ttctagctct gcatcaacgg
acaccgttac atcttcttcc tcttctagcc 240tttcagtgct accttcatct
ctttcagttt ttcaaaatcc cacagatata tcacggagca 300atcccaagtc
accacaaaaa cctatcgtta gagtcttcct gcccaataaa cagaggacgg
360tggtacccgc aagatgtgga gttacagtcc gggacagtct aaagaaagct
ctgatgatga 420gaggtctaat cccagagtgc tgtgctgttt acagaattca
ggatggagag aagaaaccga 480ttggctggga cactgatatt tcctggctca
ctggagagga attgcatgta gaagtgttgg 540aaaatgttcc gcttaccaca
cacaactttg tacggaaaac ttttttcacc ttagcatttt 600gtgacttttg
tcgaaagctg cttttccagg gttttcgctg tcaaacatgt ggttataaat
660ttcaccagcg ttgtagtaca gaggttccac tgatgtgtgt taattatgac
caacttgatt 720tgctgtttgt ctccaagttc tttgaacacc acccaatacc
acaggaggag gcctccatag 780cagagactgc ccttacgtct ggatcatccc
cttctgctcc cccctccgat tctcctgggc 840ccccaattct gaccagtccg
tctccttcaa aatccattcc aattccacag cctttccgac 900cagcagatga
agatcatcga aatcagtttg gacaacgaga ccggtcctca tcagctccaa
960atgtgcatat aaacacaata gaacccgtca acattgatga cttgattaga
gaccaagggt 1020ttcgtagtga tggaggatca accacaggtt tgtctgccac
cccccctgcc tcattgcctg 1080gctcactcac taatgtaaaa gcattacaga
aatctccagg acctcagcgg gaaagaaaat 1140catcttcatc ctcagaagat
aggaatcgaa tgaaaacact tggtagacgg gattcaagtg 1200atgattggga
gatacctgat gggcagatca cagtgggaca gagaattgga tccgggtcat
1260ttgggacagt ctacaaggga aagtggcatg gtgacgtggc agtgaaaatg
ttgaatgtga 1320cagcacccac acctcagcag ttacaggcct tcaaaaatga
agtaggagta ctcaggaaaa 1380ctcgacatgt gaatatccta ctctttatgg
gctattcaac aaagccccaa ctggctattg 1440ttacccagtg gtgtgagggc
tccagcttat atcaccatct ccacatcatt gagaccaaat 1500ttgagatgat
aaagcttata gatattgcac ggcagactgc acagggcatg gattacttac
1560acgccaagtc aatcatccac agagacctca agagtaataa tatttttctt
catgaagacc 1620tcacagtaaa aataggtgat tttggtctag ccacagtgaa
atctcgatgg agtgggtccc 1680atcagtttga acagttgtct ggatccattt
tgtggatggc accagaagtg atccgaatgc 1740aagacaaaaa cccatatagc
ttccagtcag atgtatacgc atttgggatt gttctatatg 1800aattgatgac
agggcagtta ccttattcaa acatcaacaa cagggaccag ctcagatcat
1860gatcacggtg tcatgagatc aagccccac 188914615PRTCanis familiaris
14Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu Asp Lys Phe 1
5 10 15 Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala Tyr Glu
Glu 20 25 30 Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln
Gln Leu Leu 35 40 45 Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val
Ser Ser Ser Ala Ser 50 55 60 Thr Asp Thr Val Thr Ser Ser Ser Ser
Ser Ser Leu Ser Val Leu Pro 65 70 75 80 Ser Ser Leu Ser Val Phe Gln
Asn Pro Thr Asp Ile Ser Arg Ser Asn 85 90 95 Pro Lys Ser Pro Gln
Lys Pro Ile Val Arg Val Phe Leu Pro Asn Lys 100 105 110 Gln Arg Thr
Val Val Pro Ala Arg Cys Gly Val Thr Val Arg Asp Ser 115 120 125 Leu
Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu Cys Cys Ala 130 135
140 Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly Trp Asp Thr
145 150 155 160 Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu
Val Leu Glu 165 170 175 Asn Val Pro Leu Thr Thr His Asn Phe Val Arg
Lys Thr Phe Phe Thr 180 185 190 Leu Ala Phe Cys Asp Phe Cys Arg Lys
Leu Leu Phe Gln Gly Phe Arg 195 200 205 Cys Gln Thr Cys Gly Tyr Lys
Phe His Gln Arg Cys Ser Thr Glu Val 210 215 220 Pro Leu Met Cys Val
Asn Tyr Asp Gln Leu Asp Leu Leu Phe Val Ser 225 230 235 240 Lys Phe
Phe Glu His His Pro Ile Pro Gln Glu Glu Ala Ser Ile Ala 245 250 255
Glu Thr Ala Leu Thr Ser Gly Ser Ser Pro Ser Ala Pro Pro Ser Asp 260
265 270 Ser Pro Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser Lys Ser
Ile 275 280 285 Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His
Arg Asn Gln 290 295 300 Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro
Asn Val His Ile Asn 305 310 315 320 Thr Ile Glu Pro Val Asn Ile Asp
Asp Leu Ile Arg Asp Gln Gly Phe 325 330 335 Arg Ser Asp Gly Gly Ser
Thr Thr Gly Leu Ser Ala Thr Pro Pro Ala 340 345 350 Ser Leu Pro Gly
Ser Leu Thr Asn Val Lys Ala Leu Gln Lys Ser Pro 355 360 365 Gly Pro
Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu Asp Arg Asn 370 375 380
Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp Trp Glu Ile 385
390 395 400 Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser Gly
Ser Phe 405 410 415 Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val
Ala Val Lys Met 420 425 430 Leu Asn Val Thr Ala Pro Thr Pro Gln Gln
Leu Gln Ala Phe Lys Asn 435 440 445 Glu Val Gly Val Leu Arg Lys Thr
Arg His Val Asn Ile Leu Leu Phe 450 455 460 Met Gly Tyr Ser Thr Lys
Pro Gln Leu Ala Ile Val Thr Gln Trp Cys 465 470 475 480 Glu Gly Ser
Ser Leu Tyr His His Leu His Ile Ile Glu Thr Lys Phe 485 490 495 Glu
Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala Gln Gly Met 500 505
510 Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu Lys Ser Asn
515 520 525 Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly Asp
Phe Gly 530 535 540 Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His
Gln Phe Glu Gln 545 550 555 560 Leu Ser Gly Ser Ile Leu Trp Met Ala
Pro Glu Val Ile Arg Met Gln 565 570 575 Asp Lys Asn Pro Tyr Ser Phe
Gln Ser Asp Val Tyr Ala Phe Gly Ile 580 585 590 Val Leu Tyr Glu Leu
Met Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn 595 600 605 Asn Arg Asp
Gln Leu Arg Ser 610 615 153521DNAFelis
catusmisc_feature(630)..(649)n is a, c, g, or t 15atgcctaacc
tcagtctctg ccaccacggc caatttgctc atgtgcccac tgtgtcggca 60ctgggatatt
ttgtgatttg ccttggccat tgtccactgt ccttgacatt gcctgaagga
120gaaccactga tgctaatgtt gaaggtgacc tttgcaggct ctccactact
cataccaaag 180atgcggcccc ctgataatcc cagagccact gtctgcacat
gggcaaaaca ggctacattc 240tgtgcagact ggggagaaag gttccaagaa
cacagtgcca tagttttggg cagagagttt 300caacacagca tagtgtctat
ggcagtatct ggatttggcc gggggaagtg cccaagagga 360gacagtcagg
ctgtgtccta
cggccaagga cctgcactta tttttgcatg cagtggttta 420gcacagggaa
gagaacgaag taggaaatcg gagccatgga aacggcagag cggaggaaac
480gtgcacgcgc gagggtgggc acgaaaggaa agaaccctcc ccagaagact
gcgcgagggc 540gctcctagga ttacgtcacg caccccgcga aaactgaaat
gtactgtgtg tggtctttta 600attgaactat cttccttatg tgcacttaan
nnnnnnnnnn nnnnnnnnng cggcggcggc 660ggtggcgcgg agcagggcca
ggctctgttc aacggggaca tggagcccga agccggcgcc 720gcggcctctt
cggctgcgga ccctgccatt cccgaggagg tgtggaatat caaacaaatg
780attaagttga cacaggaaca tatagaggcc ctattggaca aatttggtgg
ggagcataat 840ccaccatcaa tatatctaga ggcctatgaa gaatacacca
gcaagctaga tgccctccaa 900cagagagaac aacagttatt ggaatccctg
gggaatggaa ctgatttttc tgtttctagc 960tctgcatcaa cagacaccgt
tacatcttcc tcctcttcta gcctttcagt gctaccttca 1020tctctttcag
tttttcaaaa ccccacagat gtgtcacgga gcaatcccaa gtcaccacag
1080aaacctatcg ttagagtctt cctgcctaat aaacagagga cagtggtacc
tgcaagatgt 1140ggagttacag tccgggacag tctaaagaaa gctctgatga
tgagaggtct aatccctgag 1200tgctgtgctg tttacagaat tcaggatgga
gagaagaaac caattggctg ggacactgat 1260atctcctggc tcaccggaga
ggaattgcat gtagaagtgt tggaaaatgt tccacttaca 1320actcacaact
ttgtatgtac ggaaaacgtt ttcaccttag cattttgtga cttttgtcga
1380aagctgcttt tccaaggttt tcgctgtcaa acgtgtggtt ataaatttca
ccagcgttgt 1440agtacagagg ttccactgat gtgtgttaat tatgaccaac
ttgatttgct gtttgtctcc 1500aagttctttg aacaccaccc aataccacag
gaggaggcct ccatagcaga gactgcccta 1560acgtctggat cgtccccttc
tgcccccccc tccgattcta ctgggcccca aattctcacc 1620agtccgtctc
cttcaaaatc cattccaatt ccacagcctt tccgaccagc agatgaagat
1680catcgaaatc aatttggaca gcgagaccgg tcctcatcag ctccaaatgt
gcatataaat 1740acaatagaac ctgtcaatat tgatgacttg attagagacc
aggggtttcg tagtgatgga 1800ggatcaacca caggcttgtc tgccaccccc
cctgcctcat tgccgggctc tctcactaat 1860gtaaaagcat tacagaaatc
tccagggcct cagcgggaaa ggaaatcttc ttcatcctca 1920gaagatagga
atcgaatgaa aacacttggt agaagggatt caagtgatga ttgggagatt
1980cctgatgggc agatcacagt gggacagaga attggatccg ggtcatttgg
gacagtctac 2040aagggaaagt ggcatggtga tgtggcagtg aaaatgttga
atgtgacagc acccacacct 2100cagcagttac aggccttcaa aaatgaagta
ggagtactca ggaaaactcg gcatgtgaac 2160atcctgctct tcatgggcta
ttcaacaaag ccccagctgg ctattgtcac ccagtggtgt 2220gagggctcca
gcttatacca ccatctccac atcatcgaga ccaaattcga gatgatcaag
2280ctgatagata ttgctcggca gactgcgcag ggcatggatt acttacacgc
caagtcaatc 2340atccacagag acctcaagag taataatatt tttcttcacg
aagacctcac agtaaaaata 2400ggtgattttg gtctagccac agtgaaatct
cgatggagtg ggtcccatca gtttgaacag 2460ttgtctggat ccattttgtg
gatggcacca gaagtaattc gaatgcaaga taaaaaccca 2520tatagctttc
agtcagatgt atatgcattt gggattgttc tatatgaatt gatgactgga
2580cagttacctt attcaaacat caacaacagg gaccagataa tttttatggt
gggacgagga 2640tatctttctc cagatctcag taaggtacga agtaactgtc
caaaagccat gaagagattg 2700atggcagagt gcctaaaaaa gaaaagagat
gagaggccac tgtttcccca aattcttgcc 2760tctattgagc tgctggcccg
ctcattgcca aaaattcacc gcagtgcatc agaaccctcc 2820ttgaatcggg
ctggcttcca gacagaggat tttagtctct atgcttgtgc ttctccaaaa
2880acacccatcc aggcaggggg atatggtgcg tttcccgtcc actgagataa
gttagatgag 2940tgcgcgagtg cagggggccg gggccaagga ggtggaaatg
tgcgtgcttc tgtactaagt 3000tggatagcat cttctttttt aaaaaaagat
gaaccaaaga atgtgtatgt ttttaaagac 3060tagatataat tatttcctga
tctaaaatgt atacttagct ttggattttc aatatccaag 3120ggttttcaaa
atgcacagac attgctgaac atttgcagta cctcttctgg aggctttact
3180tcctgttaca aattggtttt gtttactggc ttatcctaat tattaaactt
caattaaact 3240tttctcctgc accttttgtt atgagctatc acatgtccct
tagggactcg caagagcagt 3300actgcccccg tgtacgggct tgcaggtaga
aaggggatga cgggttttaa cacctgtgtg 3360aggcaaggca gtccgaacag
atctcattta ggaagccacg agagttgaat aagttatttt 3420tattcttagt
attttttctg taactacttt ttattataac ttggaaaata tggatgtcct
3480ttatacacct tagcaataga ctgaatttct ttttataaat t 352116974PRTFelis
catusmisc_feature(210)..(217)Xaa can be any naturally occurring
amino acid 16Met Pro Asn Leu Ser Leu Cys His His Gly Gln Phe Ala
His Val Pro 1 5 10 15 Thr Val Ser Ala Leu Gly Tyr Phe Val Ile Cys
Leu Gly His Cys Pro 20 25 30 Leu Ser Leu Thr Leu Pro Glu Gly Glu
Pro Leu Met Leu Met Leu Lys 35 40 45 Val Thr Phe Ala Gly Ser Pro
Leu Leu Ile Pro Lys Met Arg Pro Pro 50 55 60 Asp Asn Pro Arg Ala
Thr Val Cys Thr Trp Ala Lys Gln Ala Thr Phe 65 70 75 80 Cys Ala Asp
Trp Gly Glu Arg Phe Gln Glu His Ser Ala Ile Val Leu 85 90 95 Gly
Arg Glu Phe Gln His Ser Ile Val Ser Met Ala Val Ser Gly Phe 100 105
110 Gly Arg Gly Lys Cys Pro Arg Gly Asp Ser Gln Ala Val Ser Tyr Gly
115 120 125 Gln Gly Pro Ala Leu Ile Phe Ala Cys Ser Gly Leu Ala Gln
Gly Arg 130 135 140 Glu Arg Ser Arg Lys Ser Glu Pro Trp Lys Arg Gln
Ser Gly Gly Asn 145 150 155 160 Val His Ala Arg Gly Trp Ala Arg Lys
Glu Arg Thr Leu Pro Arg Arg 165 170 175 Leu Arg Glu Gly Ala Pro Arg
Ile Thr Ser Arg Thr Pro Arg Lys Leu 180 185 190 Lys Cys Thr Val Cys
Gly Leu Leu Ile Glu Leu Ser Ser Leu Cys Ala 195 200 205 Leu Xaa Xaa
Xaa Xaa Xaa Xaa Xaa Xaa Gly Gly Gly Gly Gly Ala Glu 210 215 220 Gln
Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala 225 230
235 240 Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp
Asn 245 250 255 Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu
Ala Leu Leu 260 265 270 Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser
Ile Tyr Leu Glu Ala 275 280 285 Tyr Glu Glu Tyr Thr Ser Lys Leu Asp
Ala Leu Gln Gln Arg Glu Gln 290 295 300 Gln Leu Leu Glu Ser Leu Gly
Asn Gly Thr Asp Phe Ser Val Ser Ser 305 310 315 320 Ser Ala Ser Thr
Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser 325 330 335 Val Leu
Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser 340 345 350
Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu 355
360 365 Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr
Val 370 375 380 Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu
Ile Pro Glu 385 390 395 400 Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly
Glu Lys Lys Pro Ile Gly 405 410 415 Trp Asp Thr Asp Ile Ser Trp Leu
Thr Gly Glu Glu Leu His Val Glu 420 425 430 Val Leu Glu Asn Val Pro
Leu Thr Thr His Asn Phe Val Cys Thr Glu 435 440 445 Asn Val Phe Thr
Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe 450 455 460 Gln Gly
Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys 465 470 475
480 Ser Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu
485 490 495 Leu Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln
Glu Glu 500 505 510 Ala Ser Ile Ala Glu Thr Ala Leu Thr Ser Gly Ser
Ser Pro Ser Ala 515 520 525 Pro Pro Ser Asp Ser Thr Gly Pro Gln Ile
Leu Thr Ser Pro Ser Pro 530 535 540 Ser Lys Ser Ile Pro Ile Pro Gln
Pro Phe Arg Pro Ala Asp Glu Asp 545 550 555 560 His Arg Asn Gln Phe
Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn 565 570 575 Val His Ile
Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg 580 585 590 Asp
Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala 595 600
605 Thr Pro Pro Ala Ser Leu Pro Gly Ser Leu Thr Asn Val Lys Ala Leu
610 615 620 Gln Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser
Ser Ser 625 630 635 640 Glu Asp Arg Asn Arg Met Lys Thr Leu Gly Arg
Arg Asp Ser Ser Asp 645 650 655 Asp Trp Glu Ile Pro Asp Gly Gln Ile
Thr Val Gly Gln Arg Ile Gly 660 665 670 Ser Gly Ser Phe Gly Thr Val
Tyr Lys Gly Lys Trp His Gly Asp Val 675 680 685 Ala Val Lys Met Leu
Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln 690 695 700 Ala Phe Lys
Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn 705 710 715 720
Ile Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val 725
730 735 Thr Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile
Ile 740 745 750 Glu Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala
Arg Gln Thr 755 760 765 Ala Gln Gly Met Asp Tyr Leu His Ala Lys Ser
Ile Ile His Arg Asp 770 775 780 Leu Lys Ser Asn Asn Ile Phe Leu His
Glu Asp Leu Thr Val Lys Ile 785 790 795 800 Gly Asp Phe Gly Leu Ala
Thr Val Lys Ser Arg Trp Ser Gly Ser His 805 810 815 Gln Phe Glu Gln
Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val 820 825 830 Ile Arg
Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr 835 840 845
Ala Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr 850
855 860 Ser Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe Met Val Gly Arg
Gly 865 870 875 880 Tyr Leu Ser Pro Asp Leu Ser Lys Val Arg Ser Asn
Cys Pro Lys Ala 885 890 895 Met Lys Arg Leu Met Ala Glu Cys Leu Lys
Lys Lys Arg Asp Glu Arg 900 905 910 Pro Leu Phe Pro Gln Ile Leu Ala
Ser Ile Glu Leu Leu Ala Arg Ser 915 920 925 Leu Pro Lys Ile His Arg
Ser Ala Ser Glu Pro Ser Leu Asn Arg Ala 930 935 940 Gly Phe Gln Thr
Glu Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro Lys 945 950 955 960 Thr
Pro Ile Gln Ala Gly Gly Tyr Gly Ala Phe Pro Val His 965 970
173853DNABos taurus 17ctcagctgcg ccgggtctca caagacggtt cccgaggtgg
cccaggcgcc gtcccaccgc 60cgacgccgcc cgggccgccc gggccgtccc tccccgctgc
cccccgtcct ccgcctccgc 120ctccccccgc cctcagcctc ccttccccct
ccccgcccag cagcggtcgc tcgggcccgg 180ctctcggtta taagatggcg
gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg 240agcagggcca
ggctctgttc aacggggaca tggagcccga ggccggcgcc gcggcctctt
300cggctgcgga ccccgccatt cccgaggagg tgtggaatat caaacaaatg
attaagttga 360cacaggagca tatagaggcc ctattggaca aatttggtgg
ggagcataat ccaccatcaa 420tatatctgga ggcctatgaa gaatacacca
gcaagctaga tgccctccaa caaagagaac 480aacagttatt ggaatccctg
gggaatggaa ctgatttttc tgtttctagc tctgcatcaa 540cggacaccgt
tacatcttct tcctcttcta gcctttcagt gctgccttca tctctttcag
600tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa gtcaccacaa
aaacctatcg 660ttagagtctt cctgcccaat aaacagagga cagtggtacc
tgcacggtgt ggagtcacag 720tccgggacag cctgaagaag gcactgatga
tgagaggtct aatcccagag tgctgtgctg 780tttacagaat tcaggatggg
gagaagaaac caattggctg ggacactgat atttcctggc 840ttactggaga
ggagttgcat gtagaagtgt tggagaatgt tccacttaca acacacaact
900ttgtacggaa aacttttttc accttagcat tttgtgactt ctgtagaaag
ctgcttttcc 960agggattccg ctgtcaaaca tgtggttata aatttcacca
gcgttgtagt acagaggttc 1020cactgatgtg tgttaattat gaccaactag
atttgctgtt tgtctccaag ttctttgaac 1080accacccaat accacaggag
gaggcctcct tagcagagac tacccttcca tgtggctcat 1140ccccttctgc
acccccctcc gattctattg ggcccccaat tctcaccagt ccatctcctt
1200caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat
cgaaatcagt 1260ttggacaacg agaccggtcc tcatcagctc caaatgtgca
tataaacaca atagaacccg 1320tcaatattga tgacttgatt agagaccaag
ggtttcgtag tgatggagga tcaaccacag 1380gtttatccgc cacaccccct
gcctcattac ctggctcact ctctaatgtg aaagcattgc 1440agaaatctcc
aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc
1500gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct
gatggacaga 1560tcacagtggg acaaagaatt ggatcagggt catttgggac
agtctacaag ggaaagtggc 1620atggtgatgt ggcagtgaaa atgttgaatg
tgacagcacc cacacctcag cagttacagg 1680ccttcaaaaa tgaagtagga
gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1740tgggttattc
aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt
1800tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt
atagatattg 1860cacggcagac tgcacagggc atggattact tacacgccaa
gtcaatcatc cacagagacc 1920tcaagagtaa taatattttt cttcatgaag
acctcacagt aaaaataggt gattttggtc 1980tagccacagt gaaatctcga
tggagtgggt cccatcagtt tgaacagttg tctggatcca 2040ttttgtggat
ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt
2100cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag
ttaccttatt 2160caaatatcaa caacagggac cagataattt ttatggtggg
acgaggatat ctgtctccag 2220atctcagtaa ggtacggagt aactgtccaa
aagccatgaa gagattaatg gcagagtgcc 2280taaaaaagaa aagagatgaa
agaccactct ttccccaaat tctcgcctct attgagctgc 2340tggcccgctc
attgccaaaa attcaccgca gtgcatcaga accctccttg aatcgggctg
2400gcttccaaac agaggatttt agtctatatg cttgtgcttc tccaaaaaca
cccattcagg 2460cagggggata tggtacgttt cctgttcact gaaacaaacc
gagtgagtga cagcatgtag 2520gagggtaggg acaaaagaaa gtgaacaaat
gtttgcttat atatttgtta aattgaatag 2580gattttcttt ttctttaaag
gtgaacaaga gaacatgtgt gtttttaaag tttggatata 2640gttttcttcc
cagtctaaaa cccatagtta gcattacatt ttcaacatcg aatttttttt
2700taattcatag acattgctga aaatttataa taccttttcc agaggcttta
cttcccattc 2760caagtttgtt ttgtttactt ggttagtcta atcattaaac
tttaaacttt ccccacctac 2820cttttgctgt tagctatccc gcatccatta
ggggctccaa gaacagcact gtctgcgtgt 2880gtgtgttggc aggtgggaag
ctgatggtaa gttaggctgt gttagtgaag gtaaactgac 2940caggtctaat
taggagtcac tagaattgaa taagcttatt tttattaata ttttttctta
3000taactatttc tttttgtaat aatttagaaa atataattgt tctttattcc
cttacagcag 3060tataaattat tggtgcaggt aaccaaagat attactgagg
agtggcatgt ttgacatgag 3120tgacatggtt taactttgga tttttagtta
atatttcttt atatattaag gatgtcttac 3180acattataga agtcaaattt
actgacaaag gtattgcctc ctcttcctcc ccaaaaacac 3240agcaaaattc
tctgggaact cgtagcattg ttggttttct tttggatgac tatggttgcc
3300aaacaaccaa gtaattgatt ttttttaaat tattattgct ttagattata
ctcacctctc 3360atgatgcctg ttagcaatca cctttatcca tgtgtcttgt
aaaatatctt tcctccttat 3420attctttgcc caacaagagt ctacttgtta
tgaatgagta ctattttctt tttttgattc 3480cccagtataa ttagtatgtt
tagtgctttc taggacttcc actttcttat gttaaaaaaa 3540aaaacaaact
aatgtggcag tcagtatatt cttactgtga atcagagtct ttactgggaa
3600tcaaagtgaa agaagcagct gttctgactt cagagtcagc ctagggacca
aaaccagcct 3660cttaaataca ccttcattta ttcagtttgg atttgtgatg
attttcatta tagctgacag 3720ttcaaggtta ttcagtggca cacagatagc
atctgcataa atgcctttct tcttgaaaat 3780aaaggagaaa attgggaaga
ctttacacca atagtttagt ctttaagtac cacagataac 3840acacaccata aat
385318764PRTBos taurus 18Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly
Gly Gly Gly Ala Glu Gln 1 5 10 15 Gly Gln Ala Leu Phe Asn Gly Asp
Met Glu Pro Glu Ala Gly Ala Ala 20 25 30 Ala Ser Ser Ala Ala Asp
Pro Ala Ile Pro Glu Glu Val Trp Asn Ile 35 40 45 Lys Gln Met Ile
Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu Asp 50 55 60 Lys Phe
Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala Tyr 65 70 75 80
Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln Gln 85
90 95 Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser
Ser 100 105 110 Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser
Leu Ser Val 115 120 125 Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro
Thr Asp Val Ser Arg 130 135 140 Ser Asn Pro Lys Ser Pro Gln Lys Pro
Ile Val Arg Val Phe Leu Pro 145 150 155 160 Asn Lys Gln Arg Thr Val
Val Pro Ala Arg Cys Gly Val Thr Val Arg 165 170 175 Asp Ser Leu Lys
Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu Cys 180 185 190 Cys Ala
Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly Trp 195 200 205
Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu Val 210
215 220 Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr
Phe 225
230 235 240 Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe
Gln Gly 245 250 255 Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln
Arg Cys Ser Thr 260 265 270 Glu Val Pro Leu Met Cys Val Asn Tyr Asp
Gln Leu Asp Leu Leu Phe 275 280 285 Val Ser Lys Phe Phe Glu His His
Pro Ile Pro Gln Glu Glu Ala Ser 290 295 300 Leu Ala Glu Thr Thr Leu
Pro Cys Gly Ser Ser Pro Ser Ala Pro Pro 305 310 315 320 Ser Asp Ser
Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser Lys 325 330 335 Ser
Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His Arg 340 345
350 Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val His
355 360 365 Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg
Asp Gln 370 375 380 Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu
Ser Ala Thr Pro 385 390 395 400 Pro Ala Ser Leu Pro Gly Ser Leu Ser
Asn Val Lys Ala Leu Gln Lys 405 410 415 Ser Pro Gly Pro Gln Arg Glu
Arg Lys Ser Ser Ser Ser Ser Glu Asp 420 425 430 Arg Asn Arg Met Lys
Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp Trp 435 440 445 Glu Ile Pro
Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser Gly 450 455 460 Ser
Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala Val 465 470
475 480 Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala
Phe 485 490 495 Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val
Asn Ile Leu 500 505 510 Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu
Ala Ile Val Thr Gln 515 520 525 Trp Cys Glu Gly Ser Ser Leu Tyr His
His Leu His Ile Ile Glu Thr 530 535 540 Lys Phe Glu Met Ile Lys Leu
Ile Asp Ile Ala Arg Gln Thr Ala Gln 545 550 555 560 Gly Met Asp Tyr
Leu His Ala Lys Ser Ile Ile His Arg Asp Leu Lys 565 570 575 Ser Asn
Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly Asp 580 585 590
Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln Phe 595
600 605 Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile
Arg 610 615 620 Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val
Tyr Ala Phe 625 630 635 640 Gly Ile Val Leu Tyr Glu Leu Met Thr Gly
Gln Leu Pro Tyr Ser Asn 645 650 655 Ile Asn Asn Arg Asp Gln Ile Ile
Phe Met Val Gly Arg Gly Tyr Leu 660 665 670 Ser Pro Asp Leu Ser Lys
Val Arg Ser Asn Cys Pro Lys Ala Met Lys 675 680 685 Arg Leu Met Ala
Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro Leu 690 695 700 Phe Pro
Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser Leu Pro 705 710 715
720 Lys Ile His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg Ala Gly Phe
725 730 735 Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro Lys
Thr Pro 740 745 750 Ile Gln Ala Gly Gly Tyr Gly Thr Phe Pro Val His
755 760 19 4936DNABos taurus 19ctcagctgcg ccgggtctca caagacggtt
cccgaggtgg cccaggcgcc gtcccaccgc 60cgacgccgcc cgggccgccc gggccgtccc
tccccgctgc cccccgtcct ccgcctccgc 120ctccccccgc cctcagcctc
ccttccccct ccccgcccag cagcggtcgc tcgggcccgg 180ctctcggtta
taagatggcg gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg
240agcagggcca ggctctgttc aacggggaca tggagcccga ggccggcgcc
gcggcctctt 300cggctgcgga ccccgccatt cccgaggagg tgtggaatat
caaacaaatg attaagttga 360cacaggagca tatagaggcc ctattggaca
aatttggtgg ggagcataat ccaccatcaa 420tatatctgga ggcctatgaa
gaatacacca gcaagctaga tgccctccaa caaagagaac 480aacagttatt
ggaatccctg gggaatggaa ctgatttttc tgtttctagc tctgcatcaa
540cggacaccgt tacatcttct tcctcttcta gcctttcagt gctgccttca
tctctttcag 600tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa
gtcaccacaa aaacctatcg 660ttagagtctt cctgcccaat aaacagagga
cagtggtacc tgcacggtgt ggagtcacag 720tccgggacag cctgaagaag
gcactgatga tgagaggtct aatcccagag tgctgtgctg 780tttacagaat
tcaggatggg gagaagaaac caattggctg ggacactgat atttcctggc
840ttactggaga ggagttgcat gtagaagtgt tggagaatgt tccacttaca
acacacaact 900ttgtacggaa aacttttttc accttagcat tttgtgactt
ctgtagaaag ctgcttttcc 960agggattccg ctgtcaaaca tgtggttata
aatttcacca gcgttgtagt acagaggttc 1020cactgatgtg tgttaattat
gaccaactag atttgctgtt tgtctccaag ttctttgaac 1080accacccaat
accacaggag gaggcctcct tagcagagac tacccttcca tgtggctcat
1140ccccttctgc acccccctcc gattctattg ggcccccaat tctcaccagt
ccatctcctt 1200caaaatccat tccaattcca cagcctttcc gaccagcaga
tgaagatcat cgaaatcagt 1260ttggacaacg agaccggtcc tcatcagctc
caaatgtgca tataaacaca atagaacccg 1320tcaatattga tgacttgatt
agagaccaag ggtttcgtag tgatggagga tcaaccacag 1380gtttatccgc
cacaccccct gcctcattac ctggctcact ctctaatgtg aaagcattgc
1440agaaatctcc aggacctcag cgagaaagaa agtcctcttc atcctcagaa
gacaggaatc 1500gaatgaaaac gcttggtaga cgggattcaa gtgacgattg
ggagattcct gatggacaga 1560tcacagtggg acaaagaatt ggatcagggt
catttgggac agtctacaag ggaaagtggc 1620atggtgatgt ggcagtgaaa
atgttgaatg tgacagcacc cacacctcag cagttacagg 1680ccttcaaaaa
tgaagtagga gtactcagga aaacgcgaca tgtgaatatc ctcctcttca
1740tgggttattc aacaaagcca caactggcta ttgttaccca gtggtgtgag
ggctccagtt 1800tatatcatca tctccacatc attgagacca aattcgagat
gatcaaactt atagatattg 1860cacggcagac tgcacagggc atggattact
tacacgccaa gtcaatcatc cacagagacc 1920tcaagagtaa taatattttt
cttcatgaag acctcacagt aaaaataggt gattttggtc 1980tagccacagt
gaaatctcga tggagtgggt cccatcagtt tgaacagttg tctggatcca
2040ttttgtggat ggcaccagaa gtaatcagaa tgcaagataa aaacccatat
agctttcagt 2100cagatgtata tgcatttggg attgttctgt atgaattgat
gaccggacag ttaccttatt 2160caaatatcaa caacagggac cagataattt
ttatggtggg acgaggatat ctgtctccag 2220atctcagtaa ggtacggagt
aactgtccaa aagccatgaa gagattaatg gcagagtgcc 2280taaaaaagaa
aagagatgaa agaccactct ttccccaagt aggaaagact ctcctaagca
2340agagacaaaa ttcagaagtt atcagggaaa aagataagca gattctcgcc
tctattgagc 2400tgctggcccg ctcattgcca aaaattcacc gcagtgcatc
agaaccctcc ttgaatcggg 2460ctggcttcca aacagaggat tttagtctat
atgcttgtgc ttctccaaaa acacccattc 2520aggcaggggg atatgaagca
gatttggctc ttacatcaaa taaaaataga gtagaagttg 2580ggatttagag
atttcctgac atgcaagaag gaataagcaa gaaaaaaagg tttgttttcc
2640ccaaatcata tctattgtct tttacttcta ttttttctta aattttttgt
gatttcagag 2700acatgtagag ttttattgat acctaaacta tgagttcttt
tttttttttt tttttcatta 2760ttttgatttt tttggccaag aggcatatgg
gatcttagct tgagaaagca acaattttct 2820tgatgtcatt ttgggtgagg
gcacatattg ctgtgaacag tgtggtgata gccaccaggg 2880accaaactca
cacccgctgc attgaaaggt gaagtcttaa acactggacc agcagagaaa
2940ttcctactct atgagttctt tttgtcatcc cctccccgca ccctccaccc
ccaacctaaa 3000gtctgatgat gaaatcaaca actattccat tagaagcagt
agattctggt agcatgatct 3060ttagtttgtt agtaagattt tgtgctttgt
ggggttgtgt cgttttaagg ctaatattta 3120agtttgtcaa atagaatgct
gttcagattg taaaaatgag taataaacat ctgaagtttt 3180ttttaagtta
tttttaacat ggtatataca gttgagctta gagtttatca ttttctgata
3240ttctcttact tagtagatga attctagcca ttttttataa agatttctgt
taagcaaatc 3300ctgttttcac atgggcttcc tttaagggat tttagattct
gctggatatg gtgactgctc 3360ataagactgt tgaaaattac ttttaagatg
tattagaata cttctgaaaa aaaatagcaa 3420ccttaaaacc ataagcaaaa
gtagtaaggg tgtttataca tttctagagt ccctgtttag 3480gtaatagcct
cctatgattg tactttaaat gttttgctct ccaaggtttt agtaacttgg
3540ctttttttct aatcagtgcc aaactccccc agttttttta actttaaata
tgaggtaata 3600aatcttttac ccttccttga tcttttgact tataatacct
tggtcagttg tttcttaaaa 3660ggaatcctta aatggaaaga gacaatatca
ctgtctgcag ttctgattag tagttttatt 3720cagaatggaa aaacagatta
ttcatttttg aaaattgttc aggggtatgt tcattgttag 3780gaccttggac
tttggagtca gtgcctagct atgcattcca ggtctgccat tttctggctg
3840tgaaattttg gacaagttac ttaaccactt taaaccccag ctttaagaag
taaattaacc 3900ccagtaaatt aagaagtaat agcagccact tcgtagagtt
gttatgaggc tcagatgcag 3960tgcaaatgtg tataaagtat tcagggagtc
acctggtata ctataataga cactagaata 4020gttgccaata ttatcagcat
acaatctgag gattctgtca gccaatcatt agcaatctgt 4080tgtttgttgg
gacatgccag tgttctccag ttgaaatcag tagcaatcta aaaatggata
4140gattattcct catttaaata gtgtgttcat ataagtgatt gcttggatcc
ttatcagaag 4200ttgctgttac tgaaaaatga taaggctgac taaattgtga
tagttgtcag ttactaacca 4260actcccagaa atgaataaga ggaacctatc
tctagttcct agtagaaggt atggacaaaa 4320tagtaggtga aaaataatgt
cttgaacccc caaattaagt aagctttaaa gagtacaata 4380cctcaaaggg
tctttgcggt ttaaaatttg tatgctgaga atgatgttca ttgacatgtg
4440cctatatgta attttttgat agtttaaaag gtgaaatgaa ctacagatgg
gagaggtctg 4500aattttcttg ccttcagtca aatgtgtaat gtggacatat
tatttgacct gtgaatttta 4560tcttttaaaa aagattaatt cctgcttctt
ccttcctaat agttgcatta taataatgaa 4620aatgagttga taatttgggg
ggaaagtatt ctacaaatca accttattat tttaccattg 4680gtttctgaga
aattttgttc atttgaaccg tttatagctt gattagaatc atagcatgta
4740aaacccaact gagggattat ctgcagactt aatgtagtat tatgtaagtt
gtcttctttc 4800atttcgacct tttttgcttt tgttgttgct agatctgtag
tatgtagcta gtcacctttc 4860agcgaggttt cagcgaggct tttctgtgtc
tctaggttat ttgagataac ttttttaaaa 4920ttagctcttg tcctcc
493620797PRTBos taurus 20Met Ala Ala Leu Ser Gly Gly Gly Gly Gly
Gly Gly Gly Gly Ala Glu 1 5 10 15 Gln Gly Gln Ala Leu Phe Asn Gly
Asp Met Glu Pro Glu Ala Gly Ala 20 25 30 Ala Ala Ser Ser Ala Ala
Asp Pro Ala Ile Pro Glu Glu Val Trp Asn 35 40 45 Ile Lys Gln Met
Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu 50 55 60 Asp Lys
Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala 65 70 75 80
Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln 85
90 95 Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser
Ser 100 105 110 Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser
Ser Leu Ser 115 120 125 Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn
Pro Thr Asp Val Ser 130 135 140 Arg Ser Asn Pro Lys Ser Pro Gln Lys
Pro Ile Val Arg Val Phe Leu 145 150 155 160 Pro Asn Lys Gln Arg Thr
Val Val Pro Ala Arg Cys Gly Val Thr Val 165 170 175 Arg Asp Ser Leu
Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu 180 185 190 Cys Cys
Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly 195 200 205
Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu 210
215 220 Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys
Thr 225 230 235 240 Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys
Leu Leu Phe Gln 245 250 255 Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys
Phe His Gln Arg Cys Ser 260 265 270 Thr Glu Val Pro Leu Met Cys Val
Asn Tyr Asp Gln Leu Asp Leu Leu 275 280 285 Phe Val Ser Lys Phe Phe
Glu His His Pro Ile Pro Gln Glu Glu Ala 290 295 300 Ser Leu Ala Glu
Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro 305 310 315 320 Pro
Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser 325 330
335 Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His
340 345 350 Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro
Asn Val 355 360 365 His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp
Leu Ile Arg Asp 370 375 380 Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr
Thr Gly Leu Ser Ala Thr 385 390 395 400 Pro Pro Ala Ser Leu Pro Gly
Ser Leu Ser Asn Val Lys Ala Leu Gln 405 410 415 Lys Ser Pro Gly Pro
Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu 420 425 430 Asp Arg Asn
Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp 435 440 445 Trp
Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser 450 455
460 Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala
465 470 475 480 Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln
Leu Gln Ala 485 490 495 Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr
Arg His Val Asn Ile 500 505 510 Leu Leu Phe Met Gly Tyr Ser Thr Lys
Pro Gln Leu Ala Ile Val Thr 515 520 525 Gln Trp Cys Glu Gly Ser Ser
Leu Tyr His His Leu His Ile Ile Glu 530 535 540 Thr Lys Phe Glu Met
Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala 545 550 555 560 Gln Gly
Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu 565 570 575
Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly 580
585 590 Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His
Gln 595 600 605 Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro
Glu Val Ile 610 615 620 Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln
Ser Asp Val Tyr Ala 625 630 635 640 Phe Gly Ile Val Leu Tyr Glu Leu
Met Thr Gly Gln Leu Pro Tyr Ser 645 650 655 Asn Ile Asn Asn Arg Asp
Gln Ile Ile Phe Met Val Gly Arg Gly Tyr 660 665 670 Leu Ser Pro Asp
Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met 675 680 685 Lys Arg
Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro 690 695 700
Leu Phe Pro Gln Val Gly Lys Thr Leu Leu Ser Lys Arg Gln Asn Ser 705
710 715 720 Glu Val Ile Arg Glu Lys Asp Lys Gln Ile Leu Ala Ser Ile
Glu Leu 725 730 735 Leu Ala Arg Ser Leu Pro Lys Ile His Arg Ser Ala
Ser Glu Pro Ser 740 745 750 Leu Asn Arg Ala Gly Phe Gln Thr Glu Asp
Phe Ser Leu Tyr Ala Cys 755 760 765 Ala Ser Pro Lys Thr Pro Ile Gln
Ala Gly Gly Tyr Glu Ala Asp Leu 770 775 780 Ala Leu Thr Ser Asn Lys
Asn Arg Val Glu Val Gly Ile 785 790 795 214154DNABos taurus
21ctcagctgcg ccgggtctca caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc
60cgacgccgcc cgggccgccc gggccgtccc tccccgctgc cccccgtcct ccgcctccgc
120ctccccccgc cctcagcctc ccttccccct ccccgcccag cagcggtcgc
tcgggcccgg 180ctctcggtta taagatggcg gcgctgagtg gcggcggcgg
cggcggcggc ggtggcgcgg 240agcagggcca ggctctgttc aacggggaca
tggagcccga ggccggcgcc gcggcctctt 300cggctgcgga ccccgccatt
cccgaggagg tgtggaatat caaacaaatg attaagttga 360cacaggagca
tatagaggcc ctattggaca aatttggtgg ggagcataat ccaccatcaa
420tatatctgga ggcctatgaa gaatacacca gcaagctaga tgccctccaa
caaagagaac 480aacagttatt ggaatccctg gggaatggaa ctgatttttc
tgtttctagc tctgcatcaa 540cggacaccgt tacatcttct tcctcttcta
gcctttcagt gctgccttca tctctttcag 600tttttcaaaa tcccacagat
gtgtcacgga gcaaccccaa gtcaccacaa aaacctatcg 660ttagagtctt
cctgcccaat aaacagagga cagtggtacc tgcacggtgt ggagtcacag
720tccgggacag cctgaagaag gcactgatga tgagaggtct aatcccagag
tgctgtgctg 780tttacagaat tcaggatggg gagaagaaac caattggctg
ggacactgat atttcctggc 840ttactggaga ggagttgcat gtagaagtgt
tggagaatgt tccacttaca acacacaact 900ttgtacggaa aacttttttc
accttagcat tttgtgactt ctgtagaaag ctgcttttcc 960agggattccg
ctgtcaaaca tgtggttata aatttcacca gcgttgtagt acagaggttc
1020cactgatgtg tgttaattat gaccaactag atttgctgtt tgtctccaag
ttctttgaac 1080accacccaat accacaggag gaggcctcct tagcagagac
tacccttcca tgtggctcat 1140ccccttctgc acccccctcc gattctattg
ggcccccaat tctcaccagt ccatctcctt 1200caaaatccat tccaattcca
cagcctttcc gaccagcaga tgaagatcat cgaaatcagt 1260ttggacaacg
agaccggtcc tcatcagctc caaatgtgca tataaacaca atagaacccg
1320tcaatattga
tgacttgatt agagaccaag ggtttcgtag tgatggagga tcaaccacag
1380gtttatccgc cacaccccct gcctcattac ctggctcact ctctaatgtg
aaagcattgc 1440agaaatctcc aggacctcag cgagaaagaa agtcctcttc
atcctcagaa gacaggaatc 1500gaatgaaaac gcttggtaga cgggattcaa
gtgacgattg ggagattcct gatggacaga 1560tcacagtggg acaaagaatt
ggatcagggt catttgggac agtctacaag ggaaagtggc 1620atggtgatgt
ggcagtgaaa atgttgaatg tgacagcacc cacacctcag cagttacagg
1680ccttcaaaaa tgaagtagga gtactcagga aaacgcgaca tgtgaatatc
ctcctcttca 1740tgggttattc aacaaagcca caactggcta ttgttaccca
gtggtgtgag ggctccagtt 1800tatatcatca tctccacatc attgagacca
aattcgagat gatcaaactt atagatattg 1860cacggcagac tgcacagggc
atggattact tacacgccaa gtcaatcatc cacagagacc 1920tcaagagtaa
taatattttt cttcatgaag acctcacagt aaaaataggt gattttggtc
1980tagccacagt gaaatctcga tggagtgggt cccatcagtt tgaacagttg
tctggatcca 2040ttttgtggat ggcaccagaa gtaatcagaa tgcaagataa
aaacccatat agctttcagt 2100cagatgtata tgcatttggg attgttctgt
atgaattgat gaccggacag ttaccttatt 2160caaatatcaa caacagggac
cagataattt ttatggtggg acgaggatat ctgtctccag 2220atctcagtaa
ggtacggagt aactgtccaa aagccatgaa gagattaatg gcagagtgcc
2280taaaaaagaa aagagatgaa agaccactct ttccccaagt aggaaagact
ctcctaagca 2340agagacaaaa ttcagaagtt atcagggaaa aagataagca
gattctcgcc tctattgagc 2400tgctggcccg ctcattgcca aaaattcacc
gcagtgcatc agaaccctcc ttgaatcggg 2460ctggcttcca aacagaggat
tttagtctat atgcttgtgc ttctccaaaa acacccattc 2520aggcaggggg
atatggctga gcacattgtc catcacccac aagtggctgg ttctcatcgc
2580agaatctacg tagggaatcg ggcgtgaaat tcacttaaga gatagagcag
aggaagtgtt 2640ctgtttacag gaatggagat gagagttatg agtaagttgc
ttagtcagtt ggctttgttt 2700tgaaaattat tgtgttatat ttgtgttaac
ctacttgtgt tttgacagta tatgtcacat 2760aggaagaaac ctcagactag
cataataaca aagctcagac taggcacaga tgtacacaga 2820atggaccaaa
atgggatggg ggaaggtatg ggaataagtc taggggtagg gaaaaattga
2880tgtgagggtg ggaaataaac tgtaattacc tgaaataaaa tgtaagagtg
caataagtgt 2940gctttttatt ctaagctgtg aatgggtttt ttaaaaaaag
cattccttcc caatgcattt 3000gcctatgttc catagctgat taaaaccagc
tatataaaca tatgcctttt tattcatgtt 3060aattaccaat ataaatggct
aacctttacg tcttatttat cttcatgtta tgttagttta 3120catacaggga
tgtgtgtgtg tgtgtatgct ataaattttc cctccttcgt ttaaaaacgc
3180gtttgttgga tcctctctgt ttccttaggc catgccacag ctcatagtct
cagcttggcc 3240ttcctgtcac ctgatctgaa ggactatcac agtgacgtag
ctcgttcatt ggttgtacac 3300actctaaccc ttttccttgc tcagcaatta
ctgtgtcttc taaaacagga gtgtacaacc 3360atgagattgc aattaattgt
ttgacatatg tccctttgaa ttctatttat tagttatgat 3420tgattgctct
ttggtttgga ccaagaaaaa cgaaatccca cctccccacc ttttcactta
3480tttcttactt tgaggacaat tctgtaagag agaggaaagg gaactccttc
atgttttaac 3540tgcagcaagt taatggccct ggtttacacc aaacattatg
gtgattcaca ttcacattcc 3600tctcctctct tgctgccaga ggtttgggtt
ttgttcagtt ctgctcaagc actgaaaaag 3660ttttcatgga gtctggagag
tgcccagtga aaagatggtt tttaattgtc cacagacctt 3720tctgttcctg
ctttgcaaaa attacaaagg agtaactatt tttaaagctt atttttcaat
3780tcataaaaaa gacatttatt ttcagtcaga tgatgtctcc ttgtccctta
atcctcaatg 3840tttgcttgaa tctttttttt ttttctgatt ttctcccatc
cccacttctt gatacttctt 3900gagttctctt tcctgctcag gtcctttcat
ttgtactttg gagttttttc tcatgtaaat 3960ttgtacaatg gaaaatattg
ttcagtttgg atagaacgca tggagaatta aataaaaaag 4020atagctgaaa
ttcagattga aatttatttg tgtaaagtta tttaaaaact ctgtactata
4080taaaaggcaa aaaaagttct atgtacttga tgtgaatatg cgaatactgc
tataataaag 4140attgactgca tgga 415422781PRTBos taurus 22Met Ala Ala
Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu 1 5 10 15 Gln
Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala 20 25
30 Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn
35 40 45 Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala
Leu Leu 50 55 60 Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile
Tyr Leu Glu Ala 65 70 75 80 Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala
Leu Gln Gln Arg Glu Gln 85 90 95 Gln Leu Leu Glu Ser Leu Gly Asn
Gly Thr Asp Phe Ser Val Ser Ser 100 105 110 Ser Ala Ser Thr Asp Thr
Val Thr Ser Ser Ser Ser Ser Ser Leu Ser 115 120 125 Val Leu Pro Ser
Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser 130 135 140 Arg Ser
Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu 145 150 155
160 Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val
165 170 175 Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile
Pro Glu 180 185 190 Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys
Lys Pro Ile Gly 195 200 205 Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly
Glu Glu Leu His Val Glu 210 215 220 Val Leu Glu Asn Val Pro Leu Thr
Thr His Asn Phe Val Arg Lys Thr 225 230 235 240 Phe Phe Thr Leu Ala
Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln 245 250 255 Gly Phe Arg
Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser 260 265 270 Thr
Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu 275 280
285 Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala
290 295 300 Ser Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser
Ala Pro 305 310 315 320 Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr
Ser Pro Ser Pro Ser 325 330 335 Lys Ser Ile Pro Ile Pro Gln Pro Phe
Arg Pro Ala Asp Glu Asp His 340 345 350 Arg Asn Gln Phe Gly Gln Arg
Asp Arg Ser Ser Ser Ala Pro Asn Val 355 360 365 His Ile Asn Thr Ile
Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp 370 375 380 Gln Gly Phe
Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr 385 390 395 400
Pro Pro Ala Ser Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln 405
410 415 Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser
Glu 420 425 430 Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser
Ser Asp Asp 435 440 445 Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly
Gln Arg Ile Gly Ser 450 455 460 Gly Ser Phe Gly Thr Val Tyr Lys Gly
Lys Trp His Gly Asp Val Ala 465 470 475 480 Val Lys Met Leu Asn Val
Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala 485 490 495 Phe Lys Asn Glu
Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile 500 505 510 Leu Leu
Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr 515 520 525
Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu 530
535 540 Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr
Ala 545 550 555 560 Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile
His Arg Asp Leu 565 570 575 Lys Ser Asn Asn Ile Phe Leu His Glu Asp
Leu Thr Val Lys Ile Gly 580 585 590 Asp Phe Gly Leu Ala Thr Val Lys
Ser Arg Trp Ser Gly Ser His Gln 595 600 605 Phe Glu Gln Leu Ser Gly
Ser Ile Leu Trp Met Ala Pro Glu Val Ile 610 615 620 Arg Met Gln Asp
Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala 625 630 635 640 Phe
Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser 645 650
655 Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr
660 665 670 Leu Ser Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro Lys
Ala Met 675 680 685 Lys Arg Leu Met Ala Glu Cys Leu Lys Lys Lys Arg
Asp Glu Arg Pro 690 695 700 Leu Phe Pro Gln Val Gly Lys Thr Leu Leu
Ser Lys Arg Gln Asn Ser 705 710 715 720 Glu Val Ile Arg Glu Lys Asp
Lys Gln Ile Leu Ala Ser Ile Glu Leu 725 730 735 Leu Ala Arg Ser Leu
Pro Lys Ile His Arg Ser Ala Ser Glu Pro Ser 740 745 750 Leu Asn Arg
Ala Gly Phe Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys 755 760 765 Ala
Ser Pro Lys Thr Pro Ile Gln Ala Gly Gly Tyr Gly 770 775 780
237914DNABos taurus 23ctcagctgcg ccgggtctca caagacggtt cccgaggtgg
cccaggcgcc gtcccaccgc 60cgacgccgcc cgggccgccc gggccgtccc tccccgctgc
cccccgtcct ccgcctccgc 120ctccccccgc cctcagcctc ccttccccct
ccccgcccag cagcggtcgc tcgggcccgg 180ctctcggtta taagatggcg
gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg 240agcagggcca
ggctctgttc aacggggaca tggagcccga ggccggcgcc gcggcctctt
300cggctgcgga ccccgccatt cccgaggagg tgtggaatat caaacaaatg
attaagttga 360cacaggagca tatagaggcc ctattggaca aatttggtgg
ggagcataat ccaccatcaa 420tatatctgga ggcctatgaa gaatacacca
gcaagctaga tgccctccaa caaagagaac 480aacagttatt ggaatccctg
gggaatggaa ctgatttttc tgtttctagc tctgcatcaa 540cggacaccgt
tacatcttct tcctcttcta gcctttcagt gctgccttca tctctttcag
600tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa gtcaccacaa
aaacctatcg 660ttagagtctt cctgcccaat aaacagagga cagtggtacc
tgcacggtgt ggagtcacag 720tccgggacag cctgaagaag gcactgatga
tgagaggtct aatcccagag tgctgtgctg 780tttacagaat tcaggatggg
gagaagaaac caattggctg ggacactgat atttcctggc 840ttactggaga
ggagttgcat gtagaagtgt tggagaatgt tccacttaca acacacaact
900ttgtacggaa aacttttttc accttagcat tttgtgactt ctgtagaaag
ctgcttttcc 960agggattccg ctgtcaaaca tgtggttata aatttcacca
gcgttgtagt acagaggttc 1020cactgatgtg tgttaattat gaccaactag
atttgctgtt tgtctccaag ttctttgaac 1080accacccaat accacaggag
gaggcctcct tagcagagac tacccttcca tgtggctcat 1140ccccttctgc
acccccctcc gattctattg ggcccccaat tctcaccagt ccatctcctt
1200caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat
cgaaatcagt 1260ttggacaacg agaccggtcc tcatcagctc caaatgtgca
tataaacaca atagaacccg 1320tcaatattga tgacttgatt agagaccaag
ggtttcgtag tgatggagga tcaaccacag 1380gtttatccgc cacaccccct
gcctcattac ctggctcact ctctaatgtg aaagcattgc 1440agaaatctcc
aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc
1500gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct
gatggacaga 1560tcacagtggg acaaagaatt ggatcagggt catttgggac
agtctacaag ggaaagtggc 1620atggtgatgt ggcagtgaaa atgttgaatg
tgacagcacc cacacctcag cagttacagg 1680ccttcaaaaa tgaagtagga
gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1740tgggttattc
aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt
1800tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt
atagatattg 1860cacggcagac tgcacagggc atggattact tacacgccaa
gtcaatcatc cacagagacc 1920tcaagagtaa taatattttt cttcatgaag
acctcacagt aaaaataggt gattttggtc 1980tagccacagt gaaatctcga
tggagtgggt cccatcagtt tgaacagttg tctggatcca 2040ttttgtggat
ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt
2100cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag
ttaccttatt 2160caaatatcaa caacagggac cagataattt ttatggtggg
acgaggatat ctgtctccag 2220atctcagtaa ggtacggagt aactgtccaa
aagccatgaa gagattaatg gcagagtgcc 2280taaaaaagaa aagagatgaa
agaccactct ttccccaaat tctcgcctct attgagctgc 2340tggcccgctc
attgccaaaa attcaccgca gtgcatcaga accctccttg aatcgggctg
2400gcttccaaac agaggatttt agtctatatg cttgtgcttc tccaaaaaca
cccattcagg 2460cagggggata tggagaattt gcagccttca agtagccaca
ccatcatgac agcatctact 2520cttatttctt aagtcttgtg ttcgtacaat
ttgttaacat caaaacacag ttctgttcct 2580caactctttt taaagttaaa
atttttcagt gcataagctg gtgtggaaca gaaggaaatt 2640tcccatccaa
caaaagaggg aagaatgttt taggaaccag aattctctgc tgccagtgtt
2700tcttcttcaa cacaaatatc acaagtctgc ccactcccag gaagaaagag
gagagaccct 2760gagttctgac cttttgatgg tcaggcatga tggaaagaaa
ctgctgctac agcttgggag 2820atttgctctg ggaagtctgc cagtcaactt
tgcccttcta accaccagat caatatgtgg 2880ctgatcatct gatggggcag
ttgcaatcac caagccttgt tctctttcct gttctgggat 2940tgtgttgtgg
aacccttttc cctagccacc accagttcat ttctgaggga tggaacaaaa
3000atgcagcatg cccttcctgt gtggtgcatg ttcagtcctt gacaaatttt
taccaaaatg 3060aagctacttt atttaaaagg agggtgagag gtgaggaggt
cactttgggt gtggcggaaa 3120gggaatgctg catctttttc ctgggctgct
ggggctctgg ccttggcttg ccagccggaa 3180gcgctggcac gcatcgcctt
cttttcccat tgggtccagc aatgaagacg agtgtttggg 3240gttttttttt
tctccaccat gtagcaagtt ctcaggaaaa tacaattgat atcttcctcc
3300taagctcttc caatcagtca ccaagtactt atgtggttac tttgtccagg
gcacaaaatg 3360cctgtatcta attaaaagcc tacaaaactg cttgataaca
gttttgaatg tgagacattt 3420atgtaattta aatgtaaggt acaagtttta
atttctgagt ttcttctatt atatttttat 3480taaaaaaaga aaataatttt
cagattgaat tggagtaaaa taatattact tcccactaga 3540attatatatc
ctggaaaatt gtatttttgt tacataagca gcttttaaag aaagatcatt
3600acccttttct ctacataaat atatggggag tcttagccta atgacaaata
tttataattt 3660ttaaattaat ggtacttgct ggatccatac taacatcttt
actaatacct cattgtttct 3720tccaacttac tcctacacta catcctacat
cttcttccta gtcttttatc tagaatatgc 3780aacctcaaat aaaaatggtg
gtgtcctcat tcattctcct ccttcctttt ttcccaagcc 3840tgatcttcaa
aaggttggtt aatttggcag ctgagttcct ccccaggcag agaatagacc
3900aattttaggt gtattgggac tgagggagga tgtgtaaaga ttaacatcag
taaagaaccg 3960ctgtggagta attaagaact ttgttcttta taactggaga
atataaccta accctaacat 4020ccctcagcct ttactaaagt gtggcgtaaa
tcacagtagt agcaaagaaa gtgactctgg 4080atgtgttcct ggccagtacc
tcccttatca tgaatgtaga ctctctcatc aagatttagg 4140aatataaatc
aaatcaaatg tgcccagcca agctatgtag taagggactt gaacaatatt
4200aggcagaacc tataaaataa atcagggaat tagaaattat ttaaagtttt
caaattgtaa 4260attgccccgg tgtctttcag cctactgcca ttatttttgc
tacaatacct acatttcaga 4320ggagggccta ctgaaaattc catgcaagtg
gaaaataatc ctcaagttat taatgagttt 4380gaaaagcaat gagttcttaa
gtctttgtga gtagagcaag atcctacaaa attcagaaat 4440agtaaaaatg
gattcatgct gatttgaaga gcatctgtgt gcataatata atgctgcatc
4500tcttttaaaa gcagtctatt tttcttttta aatttgtccc catagatgct
tttgaacatg 4560aacatgctta tgttaccttt tccgaggttg ggaagagcca
ggagctctca ggcagggccc 4620cctccctcag ctgggcagga gctgctcagg
aggagctagt tatagaggaa gcttagcgtt 4680ggcattttca aaattcaagg
tgataacgct ttcttcttcc tttctgtttt agaatagatt 4740gctgtctgat
ttgaaaaagg gaaatagatt tgatctcaaa tgaatctgtg cccagaagcc
4800aggctcaggg tattcagaga tttgtatagt gccctcaaaa aataacaaaa
ttttagcttt 4860ccttttttct tcttttctcc atcaaattct tttttctcta
gtttacaaat gacatggaaa 4920aggaatttcc cctgagtttt gtatgccttt
ttttttttgg cttagactat agataggcgt 4980gttgagctcc taagaaaata
caaggaggaa ctctttgttg tgcagagcac tttatgagta 5040gtttgtgtgg
ataatatgtg actgcttccc tgacgagctt gtgaggctgt acttatgtct
5100ttcctgtaag gcagcttcag tgccttctgt agtgtatata aggaaagatt
acgccttctg 5160aaaaatctca gagcaaccat aagattattt taaaatatgt
agtatgactg atggactttt 5220tcatcattaa attagtctag catctaaact
tttaccactg aaataatatt gaccaaaaag 5280caatttataa aaggtatttg
tgaatagaaa atacaatgtg atcatttgta cttatgtgca 5340ccttaaaaga
ggaattctgt ctagctgtca aattctggtt ccttaacatc cagtccttga
5400ttgtgattga gatctggtag gacgtgctgg ggcacgctag cagataaaat
cccgtatact 5460ttaggataga tgttacattt atgtcagtgt tggcaaagag
cattgtgtag taataaagaa 5520ttcaagactt cagcaatgtc aacctgaaac
tttgtaaata tttcctagat tgttatttga 5580tgcagtcaca gctctttatc
acacaatgtt gtctttccct catcaggcaa ttttagaact 5640gctgcacacc
cctcctcaga tctcacctgc ccctcctgta cattcacctc tccagccttg
5700tgcacacctc atttagcttt agtttgaaac acattgcagg gttcaggtga
cctcttcaaa 5760aactacctcc tcagaatgag gtaatgaata gttatttatt
ttaaaatatg aaaagtcagg 5820agctctagaa tatgaagatg atctaagatt
ttaactttta tgtatacttg ttgagcactc 5880tccttttgtc ctaaagggca
ttatacattt aagcagtaat actgaaaaat gtagctcaga 5940gtaactgaat
gttgttgaaa gtggtgccag aatctgtttt aggggtacgt atcagaatct
6000taatcttaaa tcggttacat gaaattaaat agttaatggt aacacttgac
taacagatat 6060aattttaatt ttcggtaggc ttttagcaag acagtaagta
catcttcata atgagttagc 6120cacagcttca tcacatgcac agattttcct
gttgagagac tgcccagtta agagggtaga 6180atgatgaacc atttttcagg
attctcttct ttgtccaaac tggcattgtg agtgctagaa 6240tatcagcact
ttcaaactag tgattccaac tattaggcta ttaaaaagca aaacaaacca
6300aacaaaccat agccagacat gggaagttta ctatgagtat aaacagcaaa
tagcttacag 6360gtcatacatt gaaatggtgt aggtaaggcg ttagaaaaat
accttgacaa tttgccaaat 6420gatcttactg tgccttcatg atgcaataaa
aaaaaaaaaa atttagcata aatcagtgat 6480ttgtgaagag agcagccacc
ctggtctaac tcagctgtgt taatattttt tagcgtgcaa 6540tttagactgc
aaagataaat gcactaaaga gtttatagcc aaaatcacat ttaaaaaatg
6600agagaaaaca caggtaaatt ttcagtgaac aaaattattt ttttaaagta
cataatccct 6660agtatagtca gatatattta tcacatagag caaataggtt
gaaatcacaa ttcagtgaca 6720tttctagaga aactttttct actcccatag
gttcttcaaa gcatggaact tttatataac 6780agaaatgtgt gacggtcatt
ttaaattgct gtagtttggg gctgaagtac tgtgtgctgg 6840gcagcaatca
catgtattaa ctagtgagaa aggagaaatt aagatatagg acagaatttg
6900attttcttgt tcccagatta ctgctgccaa cctagacact gagtttccag
aggctgaaac 6960gtaaacttgc agctcagcaa ctgttttgca aagttagtgg
gactgtcctg cttatgctgt 7020tcaaaaatgc tctgagggcc aggtggggcc
tccaggggct
cctctctgag gggacatcag 7080actagctaac gacctggcgg gcggatgtga
accggacaca ctccatggtg tgcttcttgt 7140atcggtccct cgccaccctc
aagaaaggct tcagcgggtt ctctagacgt ctccactaag 7200gtgtgttact
aacagccatg ggttgttgag cacccgagga gtgcaatagc atctctgcat
7260gattgtatat tggcccgaag agaatgaagt ggccagtgta ctcatgttcc
atgttgctag 7320ctctggtaaa ctgaaaatac tggtaagatt tttgttttat
cagtacacta gagagtaagc 7380tttgttttgt tgtttttaga taatgttttc
acttccattt ggaaagacat ttaaattgag 7440tttcagtcct aaattttgcc
agtcatggta attagcagtt tctatcaggt atttttaagg 7500tagaagagga
tagaaacata agttctaaaa gcttaaggta accgtggttt attttaaaat
7560gtttaggggt ggttagtctc tacctcaaaa aaagtgagtg aatcttttat
ttcagcattc 7620acaagttcgg ctgttgtttt tgtaatacat ttttttttta
accttttgac ccccctttac 7680ctaagtgtca atgtagtttt attaattact
aagtcagttt cattaaaatg tttatttagc 7740agttttgact aattgcaatg
attaatatag ccagttgtgc atgaggacac agccagtgag 7800tatatctggg
ttttttttgt gatgcttttt ttcttaagac ttctgtagat ttatgaagta
7860ctcattgaaa acaactaaaa tacgtttatt cgtgttaata tggaaaaaaa aaaa
791424766PRTBos taurus 24Met Ala Ala Leu Ser Gly Gly Gly Gly Gly
Gly Gly Gly Gly Ala Glu 1 5 10 15 Gln Gly Gln Ala Leu Phe Asn Gly
Asp Met Glu Pro Glu Ala Gly Ala 20 25 30 Ala Ala Ser Ser Ala Ala
Asp Pro Ala Ile Pro Glu Glu Val Trp Asn 35 40 45 Ile Lys Gln Met
Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu 50 55 60 Asp Lys
Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala 65 70 75 80
Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln 85
90 95 Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser
Ser 100 105 110 Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser
Ser Leu Ser 115 120 125 Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn
Pro Thr Asp Val Ser 130 135 140 Arg Ser Asn Pro Lys Ser Pro Gln Lys
Pro Ile Val Arg Val Phe Leu 145 150 155 160 Pro Asn Lys Gln Arg Thr
Val Val Pro Ala Arg Cys Gly Val Thr Val 165 170 175 Arg Asp Ser Leu
Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu 180 185 190 Cys Cys
Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly 195 200 205
Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu 210
215 220 Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys
Thr 225 230 235 240 Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys
Leu Leu Phe Gln 245 250 255 Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys
Phe His Gln Arg Cys Ser 260 265 270 Thr Glu Val Pro Leu Met Cys Val
Asn Tyr Asp Gln Leu Asp Leu Leu 275 280 285 Phe Val Ser Lys Phe Phe
Glu His His Pro Ile Pro Gln Glu Glu Ala 290 295 300 Ser Leu Ala Glu
Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro 305 310 315 320 Pro
Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser 325 330
335 Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His
340 345 350 Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro
Asn Val 355 360 365 His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp
Leu Ile Arg Asp 370 375 380 Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr
Thr Gly Leu Ser Ala Thr 385 390 395 400 Pro Pro Ala Ser Leu Pro Gly
Ser Leu Ser Asn Val Lys Ala Leu Gln 405 410 415 Lys Ser Pro Gly Pro
Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu 420 425 430 Asp Arg Asn
Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp 435 440 445 Trp
Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser 450 455
460 Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala
465 470 475 480 Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln
Leu Gln Ala 485 490 495 Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr
Arg His Val Asn Ile 500 505 510 Leu Leu Phe Met Gly Tyr Ser Thr Lys
Pro Gln Leu Ala Ile Val Thr 515 520 525 Gln Trp Cys Glu Gly Ser Ser
Leu Tyr His His Leu His Ile Ile Glu 530 535 540 Thr Lys Phe Glu Met
Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala 545 550 555 560 Gln Gly
Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu 565 570 575
Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly 580
585 590 Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His
Gln 595 600 605 Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro
Glu Val Ile 610 615 620 Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln
Ser Asp Val Tyr Ala 625 630 635 640 Phe Gly Ile Val Leu Tyr Glu Leu
Met Thr Gly Gln Leu Pro Tyr Ser 645 650 655 Asn Ile Asn Asn Arg Asp
Gln Ile Ile Phe Met Val Gly Arg Gly Tyr 660 665 670 Leu Ser Pro Asp
Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met 675 680 685 Lys Arg
Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro 690 695 700
Leu Phe Pro Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser Leu 705
710 715 720 Pro Lys Ile His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg
Ala Gly 725 730 735 Phe Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys Ala
Ser Pro Lys Thr 740 745 750 Pro Ile Gln Ala Gly Gly Tyr Gly Glu Phe
Ala Ala Phe Lys 755 760 765 25 4670DNABos taurus 25ggtgtgtcat
agtgcagcag attgaatgca gaagatatga aaattcagat gtcttctgtt 60aaggtgtgga
atatcaaaca aatgattaag ttgacacagg agcatataga ggccctattg
120gacaaatttg gtggggagca taatccacca tcaatatatc tggaggccta
tgaagaatac 180accagcaagc tagatgccct ccaacaaaga gaacaacagt
tattggaatc cctggggaat 240ggaactgatt tttctgtttc tagctctgca
tcaacggaca ccgttacatc ttcttcctct 300tctagccttt cagtgctgcc
ttcatctctt tcagtttttc aaaatcccac agatgtgtca 360cggagcaacc
ccaagtcacc acaaaaacct atcgttagag tcttcctgcc caataaacag
420aggacagtgg tacctgcacg gtgtggagtc acagtccggg acagcctgaa
gaaggcactg 480atgatgagag gtctaatccc agagtgctgt gctgtttaca
gaattcagga tggggagaag 540aaaccaattg gctgggacac tgatatttcc
tggcttactg gagaggagtt gcatgtagaa 600gtgttggaga atgttccact
tacaacacac aactttgtac ggaaaacttt tttcacctta 660gcattttgtg
acttctgtag aaagctgctt ttccagggat tccgctgtca aacatgtggt
720tataaatttc accagcgttg tagtacagag gttccactga tgtgtgttaa
ttatgaccaa 780ctagatttgc tgtttgtctc caagttcttt gaacaccacc
caataccaca ggaggaggcc 840tccttagcag agactaccct tccatgtggc
tcatcccctt ctgcaccccc ctccgattct 900attgggcccc caattctcac
cagtccatct ccttcaaaat ccattccaat tccacagcct 960ttccgaccag
cagatgaaga tcatcgaaat cagtttggac aacgagaccg gtcctcatca
1020gctccaaatg tgcatataaa cacaatagaa cccgtcaata ttgatgactt
gattagagac 1080caagggtttc gtagtgatgg aggatcaacc acaggtttat
ccgccacacc ccctgcctca 1140ttacctggct cactctctaa tgtgaaagca
ttgcagaaat ctccaggacc tcagcgagaa 1200agaaagtcct cttcatcctc
agaagacagg aatcgaatga aaacgcttgg tagacgggat 1260tcaagtgacg
attgggagat tcctgatgga cagatcacag tgggacaaag aattggatca
1320gggtcatttg ggacagtcta caagggaaag tggcatggtg atgtggcagt
gaaaatgttg 1380aatgtgacag cacccacacc tcagcagtta caggccttca
aaaatgaagt aggagtactc 1440aggaaaacgc gacatgtgaa tatcctcctc
ttcatgggtt attcaacaaa gccacaactg 1500gctattgtta cccagtggtg
tgagggctcc agtttatatc atcatctcca catcattgag 1560accaaattcg
agatgatcaa acttatagat attgcacggc agactgcaca gggcatggat
1620tacttacacg ccaagtcaat catccacaga gacctcaaga gtaataatat
ttttcttcat 1680gaagacctca cagtaaaaat aggtgatttt ggtctagcca
cagtgaaatc tcgatggagt 1740gggtcccatc agtttgaaca gttgtctgga
tccattttgt ggatggcacc agaagtaatc 1800agaatgcaag ataaaaaccc
atatagcttt cagtcagatg tatatgcatt tgggattgtt 1860ctgtatgaat
tgatgaccgg acagttacct tattcaaata tcaacaacag ggaccagata
1920atttttatgg tgggacgagg atatctgtct ccagatctca gtaaggtacg
gagtaactgt 1980ccaaaagcca tgaagagatt aatggcagag tgcctaaaaa
agaaaagaga tgaaagacca 2040ctctttcccc aagtaggaaa gactctccta
agcaagagac aaaattcaga agttatcagg 2100gaaaaagata agcagattct
cgcctctatt gagctgctgg cccgctcatt gccaaaaatt 2160caccgcagtg
catcagaacc ctccttgaat cgggctggct tccaaacaga ggattttagt
2220ctatatgctt gtgcttctcc aaaaacaccc attcaggcag ggggatatga
agcagatttg 2280gctcttacat caaataaaaa tagagtagaa gttgggattt
agagatttcc tgacatgcaa 2340gaaggaataa gcaagaaaaa aaggtttgtt
ttccccaaat catatctatt gtcttttact 2400tctatttttt cttaaatttt
ttgtgatttc agagacatgt agagttttat tgatacctaa 2460actatgagtt
cttttttttt tttttttttc attattttga tttttttggc caagaggcat
2520atgggatctt agcttgagaa agcaacaatt ttcttgatgt cattttgggt
gagggcacat 2580attgctgtga acagtgtggt gatagccacc agggaccaaa
ctcacacccg ctgcattgaa 2640aggtgaagtc ttaaacactg gaccagcaga
gaaattccta ctctatgagt tctttttgtc 2700atcccctccc cgcaccctcc
acccccaacc taaagtctga tgatgaaatc aacaactatt 2760ccattagaag
cagtagattc tggtagcatg atctttagtt tgttagtaag attttgtgct
2820ttgtggggtt gtgtcgtttt aaggctaata tttaagtttg tcaaatagaa
tgctgttcag 2880attgtaaaaa tgagtaataa acatctgaag ttttttttaa
gttattttta acatggtata 2940tacagttgag cttagagttt atcattttct
gatattctct tacttagtag atgaattcta 3000gccatttttt ataaagattt
ctgttaagca aatcctgttt tcacatgggc ttcctttaag 3060ggattttaga
ttctgctgga tatggtgact gctcataaga ctgttgaaaa ttacttttaa
3120gatgtattag aatacttctg aaaaaaaata gcaaccttaa aaccataagc
aaaagtagta 3180agggtgttta tacatttcta gagtccctgt ttaggtaata
gcctcctatg attgtacttt 3240aaatgttttg ctctccaagg ttttagtaac
ttggcttttt ttctaatcag tgccaaactc 3300ccccagtttt tttaacttta
aatatgaggt aataaatctt ttacccttcc ttgatctttt 3360gacttataat
accttggtca gttgtttctt aaaaggaatc cttaaatgga aagagacaat
3420atcactgtct gcagttctga ttagtagttt tattcagaat ggaaaaacag
attattcatt 3480tttgaaaatt gttcaggggt atgttcattg ttaggacctt
ggactttgga gtcagtgcct 3540agctatgcat tccaggtctg ccattttctg
gctgtgaaat tttggacaag ttacttaacc 3600actttaaacc ccagctttaa
gaagtaaatt aaccccagta aattaagaag taatagcagc 3660cacttcgtag
agttgttatg aggctcagat gcagtgcaaa tgtgtataaa gtattcaggg
3720agtcacctgg tatactataa tagacactag aatagttgcc aatattatca
gcatacaatc 3780tgaggattct gtcagccaat cattagcaat ctgttgtttg
ttgggacatg ccagtgttct 3840ccagttgaaa tcagtagcaa tctaaaaatg
gatagattat tcctcattta aatagtgtgt 3900tcatataagt gattgcttgg
atccttatca gaagttgctg ttactgaaaa atgataaggc 3960tgactaaatt
gtgatagttg tcagttacta accaactccc agaaatgaat aagaggaacc
4020tatctctagt tcctagtaga aggtatggac aaaatagtag gtgaaaaata
atgtcttgaa 4080cccccaaatt aagtaagctt taaagagtac aatacctcaa
agggtctttg cggtttaaaa 4140tttgtatgct gagaatgatg ttcattgaca
tgtgcctata tgtaattttt tgatagttta 4200aaaggtgaaa tgaactacag
atgggagagg tctgaatttt cttgccttca gtcaaatgtg 4260taatgtggac
atattatttg acctgtgaat tttatctttt aaaaaagatt aattcctgct
4320tcttccttcc taatagttgc attataataa tgaaaatgag ttgataattt
ggggggaaag 4380tattctacaa atcaacctta ttattttacc attggtttct
gagaaatttt gttcatttga 4440accgtttata gcttgattag aatcatagca
tgtaaaaccc aactgaggga ttatctgcag 4500acttaatgta gtattatgta
agttgtcttc tttcatttcg accttttttg cttttgttgt 4560tgctagatct
gtagtatgta gctagtcacc tttcagcgag gtttcagcga ggcttttctg
4620tgtctctagg ttatttgaga taactttttt aaaattagct cttgtcctcc
467026761PRTBos taurus 26Met Lys Ile Gln Met Ser Ser Val Lys Val
Trp Asn Ile Lys Gln Met 1 5 10 15 Ile Lys Leu Thr Gln Glu His Ile
Glu Ala Leu Leu Asp Lys Phe Gly 20 25 30 Gly Glu His Asn Pro Pro
Ser Ile Tyr Leu Glu Ala Tyr Glu Glu Tyr 35 40 45 Thr Ser Lys Leu
Asp Ala Leu Gln Gln Arg Glu Gln Gln Leu Leu Glu 50 55 60 Ser Leu
Gly Asn Gly Thr Asp Phe Ser Val Ser Ser Ser Ala Ser Thr 65 70 75 80
Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser Val Leu Pro Ser 85
90 95 Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser Arg Ser Asn
Pro 100 105 110 Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu Pro
Asn Lys Gln 115 120 125 Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr
Val Arg Asp Ser Leu 130 135 140 Lys Lys Ala Leu Met Met Arg Gly Leu
Ile Pro Glu Cys Cys Ala Val 145 150 155 160 Tyr Arg Ile Gln Asp Gly
Glu Lys Lys Pro Ile Gly Trp Asp Thr Asp 165 170 175 Ile Ser Trp Leu
Thr Gly Glu Glu Leu His Val Glu Val Leu Glu Asn 180 185 190 Val Pro
Leu Thr Thr His Asn Phe Val Arg Lys Thr Phe Phe Thr Leu 195 200 205
Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln Gly Phe Arg Cys 210
215 220 Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser Thr Glu Val
Pro 225 230 235 240 Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu
Phe Val Ser Lys 245 250 255 Phe Phe Glu His His Pro Ile Pro Gln Glu
Glu Ala Ser Leu Ala Glu 260 265 270 Thr Thr Leu Pro Cys Gly Ser Ser
Pro Ser Ala Pro Pro Ser Asp Ser 275 280 285 Ile Gly Pro Pro Ile Leu
Thr Ser Pro Ser Pro Ser Lys Ser Ile Pro 290 295 300 Ile Pro Gln Pro
Phe Arg Pro Ala Asp Glu Asp His Arg Asn Gln Phe 305 310 315 320 Gly
Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val His Ile Asn Thr 325 330
335 Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp Gln Gly Phe Arg
340 345 350 Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr Pro Pro
Ala Ser 355 360 365 Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln
Lys Ser Pro Gly 370 375 380 Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser
Ser Glu Asp Arg Asn Arg 385 390 395 400 Met Lys Thr Leu Gly Arg Arg
Asp Ser Ser Asp Asp Trp Glu Ile Pro 405 410 415 Asp Gly Gln Ile Thr
Val Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly 420 425 430 Thr Val Tyr
Lys Gly Lys Trp His Gly Asp Val Ala Val Lys Met Leu 435 440 445 Asn
Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu 450 455
460 Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile Leu Leu Phe Met
465 470 475 480 Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr Gln
Trp Cys Glu 485 490 495 Gly Ser Ser Leu Tyr His His Leu His Ile Ile
Glu Thr Lys Phe Glu 500 505 510 Met Ile Lys Leu Ile Asp Ile Ala Arg
Gln Thr Ala Gln Gly Met Asp 515 520 525 Tyr Leu His Ala Lys Ser Ile
Ile His Arg Asp Leu Lys Ser Asn Asn 530 535 540 Ile Phe Leu His Glu
Asp Leu Thr Val Lys Ile Gly Asp Phe Gly Leu 545 550 555 560 Ala Thr
Val Lys Ser Arg Trp Ser Gly Ser His Gln Phe Glu Gln Leu 565 570 575
Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile Arg Met Gln Asp 580
585 590 Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala Phe Gly Ile
Val 595 600 605 Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser Asn
Ile Asn Asn 610 615 620 Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly
Tyr Leu Ser Pro Asp 625 630 635 640 Leu Ser Lys Val Arg Ser Asn Cys
Pro Lys Ala Met Lys Arg Leu Met 645 650 655 Ala Glu Cys Leu Lys Lys
Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln 660 665 670 Val Gly Lys Thr
Leu Leu Ser Lys Arg Gln Asn Ser Glu Val Ile Arg 675
680 685 Glu Lys Asp Lys Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg
Ser 690 695 700 Leu Pro Lys Ile His Arg Ser Ala Ser Glu Pro Ser Leu
Asn Arg Ala 705 710 715 720 Gly Phe Gln Thr Glu Asp Phe Ser Leu Tyr
Ala Cys Ala Ser Pro Lys 725 730 735 Thr Pro Ile Gln Ala Gly Gly Tyr
Glu Ala Asp Leu Ala Leu Thr Ser 740 745 750 Asn Lys Asn Arg Val Glu
Val Gly Ile 755 760 27 4816DNABos taurus 27ctcagctgcg ccgggtctca
caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc 60cgacgccgcc cgggccgccc
gggccgtccc tccccgctgc cccccgtcct ccgcctccgc 120ctccccccgc
cctcagcctc ccttccccct ccccgcccag cagcggtcgc tcgggcccgg
180ctctcggtta taagatggcg gcgctgagtg gcggcggcgg cggcggcggc
ggtggcgcgg 240agcagggcca ggctctgttc aacggggaca tggagcccga
ggccggcgcc gcggcctctt 300cggctgcgga ccccgccatt cccgaggagg
tgtggaatat caaacaaatg attaagttga 360cacaggagca tatagaggcc
ctattggaca aatttggtgg ggagcataat ccaccatcaa 420tatatctgga
ggcctatgaa gaatacacca gcaagctaga tgccctccaa caaagagaac
480aacagttatt ggaatccctg gggaatggaa ctgatttttc tgtttctagc
tctgcatcaa 540cggacaccgt tacatcttct tcctcttcta gcctttcagt
gctgccttca tctctttcag 600tttttcaaaa tcccacagat gtgtcacgga
gcaaccccaa gtcaccacaa aaacctatcg 660ttagagtctt cctgcccaat
aaacagagga cagtggtacc tgcacggtgt ggagtcacag 720tccgggacag
cctgaagaag gcactgatga tgagaggtct aatcccagag tgctgtgctg
780tttacagaat tcaggatggg gagaagaaac caattggctg ggacactgat
atttcctggc 840ttactggaga ggagttgcat gtagaagtgt tggagaatgt
tccacttaca acacacaact 900ttgtacggaa aacttttttc accttagcat
tttgtgactt ctgtagaaag ctgcttttcc 960agggattccg ctgtcaaaca
tgtggttata aatttcacca gcgttgtagt acagaggttc 1020cactgatgtg
tgttaattat gaccaactag agcccccaat tctcaccagt ccatctcctt
1080caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat
cgaaatcagt 1140ttggacaacg agaccggtcc tcatcagctc caaatgtgca
tataaacaca atagaacccg 1200tcaatattga tgacttgatt agagaccaag
ggtttcgtag tgatggagga tcaaccacag 1260gtttatccgc cacaccccct
gcctcattac ctggctcact ctctaatgtg aaagcattgc 1320agaaatctcc
aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc
1380gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct
gatggacaga 1440tcacagtggg acaaagaatt ggatcagggt catttgggac
agtctacaag ggaaagtggc 1500atggtgatgt ggcagtgaaa atgttgaatg
tgacagcacc cacacctcag cagttacagg 1560ccttcaaaaa tgaagtagga
gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1620tgggttattc
aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt
1680tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt
atagatattg 1740cacggcagac tgcacagggc atggattact tacacgccaa
gtcaatcatc cacagagacc 1800tcaagagtaa taatattttt cttcatgaag
acctcacagt aaaaataggt gattttggtc 1860tagccacagt gaaatctcga
tggagtgggt cccatcagtt tgaacagttg tctggatcca 1920ttttgtggat
ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt
1980cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag
ttaccttatt 2040caaatatcaa caacagggac cagataattt ttatggtggg
acgaggatat ctgtctccag 2100atctcagtaa ggtacggagt aactgtccaa
aagccatgaa gagattaatg gcagagtgcc 2160taaaaaagaa aagagatgaa
agaccactct ttccccaagt aggaaagact ctcctaagca 2220agagacaaaa
ttcagaagtt atcagggaaa aagataagca gattctcgcc tctattgagc
2280tgctggcccg ctcattgcca aaaattcacc gcagtgcatc agaaccctcc
ttgaatcggg 2340ctggcttcca aacagaggat tttagtctat atgcttgtgc
ttctccaaaa acacccattc 2400aggcaggggg atatgaagca gatttggctc
ttacatcaaa taaaaataga gtagaagttg 2460ggatttagag atttcctgac
atgcaagaag gaataagcaa gaaaaaaagg tttgttttcc 2520ccaaatcata
tctattgtct tttacttcta ttttttctta aattttttgt gatttcagag
2580acatgtagag ttttattgat acctaaacta tgagttcttt tttttttttt
tttttcatta 2640ttttgatttt tttggccaag aggcatatgg gatcttagct
tgagaaagca acaattttct 2700tgatgtcatt ttgggtgagg gcacatattg
ctgtgaacag tgtggtgata gccaccaggg 2760accaaactca cacccgctgc
attgaaaggt gaagtcttaa acactggacc agcagagaaa 2820ttcctactct
atgagttctt tttgtcatcc cctccccgca ccctccaccc ccaacctaaa
2880gtctgatgat gaaatcaaca actattccat tagaagcagt agattctggt
agcatgatct 2940ttagtttgtt agtaagattt tgtgctttgt ggggttgtgt
cgttttaagg ctaatattta 3000agtttgtcaa atagaatgct gttcagattg
taaaaatgag taataaacat ctgaagtttt 3060ttttaagtta tttttaacat
ggtatataca gttgagctta gagtttatca ttttctgata 3120ttctcttact
tagtagatga attctagcca ttttttataa agatttctgt taagcaaatc
3180ctgttttcac atgggcttcc tttaagggat tttagattct gctggatatg
gtgactgctc 3240ataagactgt tgaaaattac ttttaagatg tattagaata
cttctgaaaa aaaatagcaa 3300ccttaaaacc ataagcaaaa gtagtaaggg
tgtttataca tttctagagt ccctgtttag 3360gtaatagcct cctatgattg
tactttaaat gttttgctct ccaaggtttt agtaacttgg 3420ctttttttct
aatcagtgcc aaactccccc agttttttta actttaaata tgaggtaata
3480aatcttttac ccttccttga tcttttgact tataatacct tggtcagttg
tttcttaaaa 3540ggaatcctta aatggaaaga gacaatatca ctgtctgcag
ttctgattag tagttttatt 3600cagaatggaa aaacagatta ttcatttttg
aaaattgttc aggggtatgt tcattgttag 3660gaccttggac tttggagtca
gtgcctagct atgcattcca ggtctgccat tttctggctg 3720tgaaattttg
gacaagttac ttaaccactt taaaccccag ctttaagaag taaattaacc
3780ccagtaaatt aagaagtaat agcagccact tcgtagagtt gttatgaggc
tcagatgcag 3840tgcaaatgtg tataaagtat tcagggagtc acctggtata
ctataataga cactagaata 3900gttgccaata ttatcagcat acaatctgag
gattctgtca gccaatcatt agcaatctgt 3960tgtttgttgg gacatgccag
tgttctccag ttgaaatcag tagcaatcta aaaatggata 4020gattattcct
catttaaata gtgtgttcat ataagtgatt gcttggatcc ttatcagaag
4080ttgctgttac tgaaaaatga taaggctgac taaattgtga tagttgtcag
ttactaacca 4140actcccagaa atgaataaga ggaacctatc tctagttcct
agtagaaggt atggacaaaa 4200tagtaggtga aaaataatgt cttgaacccc
caaattaagt aagctttaaa gagtacaata 4260cctcaaaggg tctttgcggt
ttaaaatttg tatgctgaga atgatgttca ttgacatgtg 4320cctatatgta
attttttgat agtttaaaag gtgaaatgaa ctacagatgg gagaggtctg
4380aattttcttg ccttcagtca aatgtgtaat gtggacatat tatttgacct
gtgaatttta 4440tcttttaaaa aagattaatt cctgcttctt ccttcctaat
agttgcatta taataatgaa 4500aatgagttga taatttgggg ggaaagtatt
ctacaaatca accttattat tttaccattg 4560gtttctgaga aattttgttc
atttgaaccg tttatagctt gattagaatc atagcatgta 4620aaacccaact
gagggattat ctgcagactt aatgtagtat tatgtaagtt gtcttctttc
4680atttcgacct tttttgcttt tgttgttgct agatctgtag tatgtagcta
gtcacctttc 4740agcgaggttt cagcgaggct tttctgtgtc tctaggttat
ttgagataac ttttttaaaa 4800ttagctcttg tcctcc 481628757PRTBos taurus
28Met Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu 1
5 10 15 Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly
Ala 20 25 30 Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu
Val Trp Asn 35 40 45 Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His
Ile Glu Ala Leu Leu 50 55 60 Asp Lys Phe Gly Gly Glu His Asn Pro
Pro Ser Ile Tyr Leu Glu Ala 65 70 75 80 Tyr Glu Glu Tyr Thr Ser Lys
Leu Asp Ala Leu Gln Gln Arg Glu Gln 85 90 95 Gln Leu Leu Glu Ser
Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser 100 105 110 Ser Ala Ser
Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser 115 120 125 Val
Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser 130 135
140 Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu
145 150 155 160 Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly
Val Thr Val 165 170 175 Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg
Gly Leu Ile Pro Glu 180 185 190 Cys Cys Ala Val Tyr Arg Ile Gln Asp
Gly Glu Lys Lys Pro Ile Gly 195 200 205 Trp Asp Thr Asp Ile Ser Trp
Leu Thr Gly Glu Glu Leu His Val Glu 210 215 220 Val Leu Glu Asn Val
Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr 225 230 235 240 Phe Phe
Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln 245 250 255
Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser 260
265 270 Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Glu Pro
Pro 275 280 285 Ile Leu Thr Ser Pro Ser Pro Ser Lys Ser Ile Pro Ile
Pro Gln Pro 290 295 300 Phe Arg Pro Ala Asp Glu Asp His Arg Asn Gln
Phe Gly Gln Arg Asp 305 310 315 320 Arg Ser Ser Ser Ala Pro Asn Val
His Ile Asn Thr Ile Glu Pro Val 325 330 335 Asn Ile Asp Asp Leu Ile
Arg Asp Gln Gly Phe Arg Ser Asp Gly Gly 340 345 350 Ser Thr Thr Gly
Leu Ser Ala Thr Pro Pro Ala Ser Leu Pro Gly Ser 355 360 365 Leu Ser
Asn Val Lys Ala Leu Gln Lys Ser Pro Gly Pro Gln Arg Glu 370 375 380
Arg Lys Ser Ser Ser Ser Ser Glu Asp Arg Asn Arg Met Lys Thr Leu 385
390 395 400 Gly Arg Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro Asp Gly
Gln Ile 405 410 415 Thr Val Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly
Thr Val Tyr Lys 420 425 430 Gly Lys Trp His Gly Asp Val Ala Val Lys
Met Leu Asn Val Thr Ala 435 440 445 Pro Thr Pro Gln Gln Leu Gln Ala
Phe Lys Asn Glu Val Gly Val Leu 450 455 460 Arg Lys Thr Arg His Val
Asn Ile Leu Leu Phe Met Gly Tyr Ser Thr 465 470 475 480 Lys Pro Gln
Leu Ala Ile Val Thr Gln Trp Cys Glu Gly Ser Ser Leu 485 490 495 Tyr
His His Leu His Ile Ile Glu Thr Lys Phe Glu Met Ile Lys Leu 500 505
510 Ile Asp Ile Ala Arg Gln Thr Ala Gln Gly Met Asp Tyr Leu His Ala
515 520 525 Lys Ser Ile Ile His Arg Asp Leu Lys Ser Asn Asn Ile Phe
Leu His 530 535 540 Glu Asp Leu Thr Val Lys Ile Gly Asp Phe Gly Leu
Ala Thr Val Lys 545 550 555 560 Ser Arg Trp Ser Gly Ser His Gln Phe
Glu Gln Leu Ser Gly Ser Ile 565 570 575 Leu Trp Met Ala Pro Glu Val
Ile Arg Met Gln Asp Lys Asn Pro Tyr 580 585 590 Ser Phe Gln Ser Asp
Val Tyr Ala Phe Gly Ile Val Leu Tyr Glu Leu 595 600 605 Met Thr Gly
Gln Leu Pro Tyr Ser Asn Ile Asn Asn Arg Asp Gln Ile 610 615 620 Ile
Phe Met Val Gly Arg Gly Tyr Leu Ser Pro Asp Leu Ser Lys Val 625 630
635 640 Arg Ser Asn Cys Pro Lys Ala Met Lys Arg Leu Met Ala Glu Cys
Leu 645 650 655 Lys Lys Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln Val
Gly Lys Thr 660 665 670 Leu Leu Ser Lys Arg Gln Asn Ser Glu Val Ile
Arg Glu Lys Asp Lys 675 680 685 Gln Ile Leu Ala Ser Ile Glu Leu Leu
Ala Arg Ser Leu Pro Lys Ile 690 695 700 His Arg Ser Ala Ser Glu Pro
Ser Leu Asn Arg Ala Gly Phe Gln Thr 705 710 715 720 Glu Asp Phe Ser
Leu Tyr Ala Cys Ala Ser Pro Lys Thr Pro Ile Gln 725 730 735 Ala Gly
Gly Tyr Glu Ala Asp Leu Ala Leu Thr Ser Asn Lys Asn Arg 740 745 750
Val Glu Val Gly Ile 755 29 2499DNABos taurus 29ctcagctgcg
ccgggtctca caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc 60cgacgccgcc
cgggccgccc gggccgtccc tccccgctgc cccccgtcct ccgcctccgc
120ctccccccgc cctcagcctc ccttccccct ccccgcccag cagcggtcgc
tcgggcccgg 180ctctcggtta taagatggcg gcgctgagtg gcggcggcgg
cggcggcggc ggtggcgcgg 240agcagggcca ggctctgttc aacggggaca
tggagcccga ggccggcgcc gcggcctctt 300cggctgcgga ccccgccatt
cccgaggagg tgtggaatat caaacaaatg attaagttga 360cacaggagca
tatagaggcc ctattggaca aatttggtgg ggagcataat ccaccatcaa
420tatatctgga ggcctatgaa gaatacacca gcaagctaga tgccctccaa
caaagagaac 480aacagttatt ggaatccctg gggaatggaa ctgatttttc
tgtttctagc tctgcatcaa 540cggacaccgt tacatcttct tcctcttcta
gcctttcagt gctgccttca tctctttcag 600tttttcaaaa tcccacagat
gtgtcacgga gcaaccccaa gtcaccacaa aaacctatcg 660ttagagtctt
cctgcccaat aaacagagga cagtggtacc tgcacggtgt ggagtcacag
720tccgggacag cctgaagaag gcactgatga tgagaggtct aatcccagag
tgctgtgctg 780tttacagaat tcaggatggg gagaagaaac caattggctg
ggacactgat atttcctggc 840ttactggaga ggagttgcat gtagaagtgt
tggagaatgt tccacttaca acacacaact 900ttgtacggaa aacttttttc
accttagcat tttgtgactt ctgtagaaag ctgcttttcc 960agggattccg
ctgtcaaaca tgtggttata aatttcacca gcgttgtagt acagaggttc
1020cactgatgtg tgttaattat gaccaactag atttgctgtt tgtctccaag
ttctttgaac 1080accacccaat accacaggag gaggcctcct tagcagagac
tacccttcca tgtggctcat 1140ccccttctgc acccccctcc gattctattg
ggcccccaat tctcaccagt ccatctcctt 1200caaaatccat tccaattcca
cagcctttcc gaccagcaga tgaagatcat cgaaatcagt 1260ttggacaacg
agaccggtcc tcatcagctc caaatgtgca tataaacaca atagaacccg
1320tcaatattga tgacttgatt agagaccaag ggtttcgtag tgatggagga
tcaaccacag 1380gtttatccgc cacaccccct gcctcattac ctggctcact
ctctaatgtg aaagcattgc 1440agaaatctcc aggacctcag cgagaaagaa
agtcctcttc atcctcagaa gacaggaatc 1500gaatgaaaac gcttggtaga
cgggattcaa gtgacgattg ggagattcct gatggacaga 1560tcacagtggg
acaaagaatt ggatcagggt catttgggac agtctacaag ggaaagtggc
1620atggtgatgt ggcagtgaaa atgttgaatg tgacagcacc cacacctcag
cagttacagg 1680ccttcaaaaa tgaagtagga gtactcagga aaacgcgaca
tgtgaatatc ctcctcttca 1740tgggttattc aacaaagcca caactggcta
ttgttaccca gtggtgtgag ggctccagtt 1800tatatcatca tctccacatc
attgagacca aattcgagat gatcaaactt atagatattg 1860cacggcagac
tgcacagggc atggattact tacacgccaa gtcaatcatc cacagagacc
1920tcaagagtaa taatattttt cttcatgaag acctcacagt aaaaataggt
gattttggtc 1980tagccacagt gaaatctcga tggagtgggt cccatcagtt
tgaacagttg tctggatcca 2040ttttgtggat ggcaccagaa gtaatcagaa
tgcaagataa aaacccatat agctttcagt 2100cagatgtata tgcatttggg
attgttctgt atgaattgat gaccggacag ttaccttatt 2160caaatatcaa
caacagggac cagataattt ttatggtggg acgaggatat ctgtctccag
2220atctcagtaa ggtacggagt aactgtccaa aagccatgaa gagattaatg
gcagagtgcc 2280taaaaaagaa aagagatgaa agaccactct ttccccaagt
aggaaagact ctcctaagca 2340agagacaaaa ttcagaagtt atcagggaaa
aagataagca ggaaaagtat gtttctttag 2400tacattccag gcatttggga
ttacagtaaa aacaatattc tcgcctctat tgagctgctg 2460gcccgctcat
tgccaaaaat tcaccgcagt gcatcagaa 249930744PRTBos taurus 30Met Ala
Ala Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu 1 5 10 15
Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala 20
25 30 Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp
Asn 35 40 45 Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu
Ala Leu Leu 50 55 60 Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser
Ile Tyr Leu Glu Ala 65 70 75 80 Tyr Glu Glu Tyr Thr Ser Lys Leu Asp
Ala Leu Gln Gln Arg Glu Gln 85 90 95 Gln Leu Leu Glu Ser Leu Gly
Asn Gly Thr Asp Phe Ser Val Ser Ser 100 105 110 Ser Ala Ser Thr Asp
Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser 115 120 125 Val Leu Pro
Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser 130 135 140 Arg
Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu 145 150
155 160 Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr
Val 165 170 175 Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu
Ile Pro Glu 180 185 190 Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu
Lys Lys Pro Ile Gly 195 200 205 Trp Asp Thr Asp Ile Ser Trp Leu Thr
Gly Glu Glu Leu His Val Glu 210 215 220 Val Leu Glu Asn Val Pro Leu
Thr Thr His Asn Phe Val Arg Lys Thr 225 230 235 240 Phe Phe Thr Leu
Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln 245 250 255 Gly Phe
Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser 260 265 270
Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu 275
280 285 Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu
Ala 290 295 300 Ser Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro
Ser Ala Pro 305 310 315 320 Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu
Thr Ser Pro Ser Pro Ser
325 330 335 Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu
Asp His 340 345 350 Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser
Ala Pro Asn Val 355 360 365 His Ile Asn Thr Ile Glu Pro Val Asn Ile
Asp Asp Leu Ile Arg Asp 370 375 380 Gln Gly Phe Arg Ser Asp Gly Gly
Ser Thr Thr Gly Leu Ser Ala Thr 385 390 395 400 Pro Pro Ala Ser Leu
Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln 405 410 415 Lys Ser Pro
Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu 420 425 430 Asp
Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp 435 440
445 Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser
450 455 460 Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp
Val Ala 465 470 475 480 Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro
Gln Gln Leu Gln Ala 485 490 495 Phe Lys Asn Glu Val Gly Val Leu Arg
Lys Thr Arg His Val Asn Ile 500 505 510 Leu Leu Phe Met Gly Tyr Ser
Thr Lys Pro Gln Leu Ala Ile Val Thr 515 520 525 Gln Trp Cys Glu Gly
Ser Ser Leu Tyr His His Leu His Ile Ile Glu 530 535 540 Thr Lys Phe
Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala 545 550 555 560
Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu 565
570 575 Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile
Gly 580 585 590 Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly
Ser His Gln 595 600 605 Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met
Ala Pro Glu Val Ile 610 615 620 Arg Met Gln Asp Lys Asn Pro Tyr Ser
Phe Gln Ser Asp Val Tyr Ala 625 630 635 640 Phe Gly Ile Val Leu Tyr
Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser 645 650 655 Asn Ile Asn Asn
Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr 660 665 670 Leu Ser
Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met 675 680 685
Lys Arg Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro 690
695 700 Leu Phe Pro Gln Val Gly Lys Thr Leu Leu Ser Lys Arg Gln Asn
Ser 705 710 715 720 Glu Val Ile Arg Glu Lys Asp Lys Gln Glu Lys Tyr
Val Ser Leu Val 725 730 735 His Ser Arg His Leu Gly Leu Gln 740 31
2404DNABos taurus 31ctcagctgcg ccgggtctca caagacggtt cccgaggtgg
cccaggcgcc gtcccaccgc 60cgacgccgcc cgggccgccc gggccgtccc tccccgctgc
cccccgtcct ccgcctccgc 120ctccccccgc cctcagcctc ccttccccct
ccccgcccag cagcggtcgc tcgggcccgg 180ctctcggtta taagatggcg
gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg 240agcagggcca
ggctctgttc aacggggaca tggagcccga ggccggcgcc gcggcctctt
300cggctgcgga ccccgccatt cccgaggagg tgtggaatat caaacaaatg
attaagttga 360cacaggagca tatagaggcc ctattggaca aatttggtgg
ggagcataat ccaccatcaa 420tatatctgga ggcctatgaa gaatacacca
gcaagctaga tgccctccaa caaagagaac 480aacagttatt ggaatccctg
gggaatggaa ctgatttttc tgtttctagc tctgcatcaa 540cggacaccgt
tacatcttct tcctcttcta gcctttcagt gctgccttca tctctttcag
600tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa gtcaccacaa
aaacctatcg 660ttagagtctt cctgcccaat aaacagagga cagtggtacc
tgcacggtgt ggagtcacag 720tccgggacag cctgaagaag gcactgatga
tgagaggtct aatcccagag tgctgtgctg 780tttacagaat tcaggatggg
gagaagaaac caattggctg ggacactgat atttcctggc 840ttactggaga
ggagttgcat gtagaagtgt tggagaatgt tccacttaca acacacaact
900ttgtacggaa aacttttttc accttagcat tttgtgactt ctgtagaaag
ctgcttttcc 960agggattccg ctgtcaaaca tgtggttata aatttcacca
gcgttgtagt acagaggttc 1020cactgatgtg tgttaattat gaccaactag
atttgctgtt tgtctccaag ttctttgaac 1080accacccaat accacaggag
gaggcctcct tagcagagac tacccttcca tgtggctcat 1140ccccttctgc
acccccctcc gattctattg ggcccccaat tctcaccagt ccatctcctt
1200caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat
cgaaatcagt 1260ttggacaacg agaccggtcc tcatcagctc caaatgtgca
tataaacaca atagaacccg 1320tcaatattga tgacttgatt agagaccaag
ggtttcgtag tgatggagga tcaaccacag 1380gtttatccgc cacaccccct
gcctcattac ctggctcact ctctaatgtg aaagcattgc 1440agaaatctcc
aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc
1500gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct
gatggacaga 1560tcacagtggg acaaagaatt ggatcagggt catttgggac
agtctacaag ggaaagtggc 1620atggtgatgt ggcagtgaaa atgttgaatg
tgacagcacc cacacctcag cagttacagg 1680ccttcaaaaa tgaagtagga
gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1740tgggttattc
aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt
1800tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt
atagatattg 1860cacggcagac tgcacagggc atggattact tacacgccaa
gtcaatcatc cacagagacc 1920tcaagagtaa taatattttt cttcatgaag
acctcacagt aaaaataggt gattttggtc 1980tagccacagt gaaatctcga
tggagtgggt cccatcagtt tgaacagttg tctggatcca 2040ttttgtggat
ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt
2100cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag
ttaccttatt 2160caaatatcaa caacagggac cagataattt ttatggtggg
acgaggatat ctgtctccag 2220atctcagtaa ggtacggagt aactgtccaa
aagccatgaa gagattaatg gcagagtgcc 2280taaaaaagaa aagagatgaa
agaccactct ttccccaaga tctctcttcc caccatagac 2340acaaaaattt
cagatggcta caggtttaca tgtaaaaaac agaattataa caaatgattt 2400ttat
240432726PRTBos taurus 32Met Ala Ala Leu Ser Gly Gly Gly Gly Gly
Gly Gly Gly Gly Ala Glu 1 5 10 15 Gln Gly Gln Ala Leu Phe Asn Gly
Asp Met Glu Pro Glu Ala Gly Ala 20 25 30 Ala Ala Ser Ser Ala Ala
Asp Pro Ala Ile Pro Glu Glu Val Trp Asn 35 40 45 Ile Lys Gln Met
Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu 50 55 60 Asp Lys
Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala 65 70 75 80
Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln 85
90 95 Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser
Ser 100 105 110 Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser
Ser Leu Ser 115 120 125 Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn
Pro Thr Asp Val Ser 130 135 140 Arg Ser Asn Pro Lys Ser Pro Gln Lys
Pro Ile Val Arg Val Phe Leu 145 150 155 160 Pro Asn Lys Gln Arg Thr
Val Val Pro Ala Arg Cys Gly Val Thr Val 165 170 175 Arg Asp Ser Leu
Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu 180 185 190 Cys Cys
Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly 195 200 205
Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu 210
215 220 Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys
Thr 225 230 235 240 Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys
Leu Leu Phe Gln 245 250 255 Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys
Phe His Gln Arg Cys Ser 260 265 270 Thr Glu Val Pro Leu Met Cys Val
Asn Tyr Asp Gln Leu Asp Leu Leu 275 280 285 Phe Val Ser Lys Phe Phe
Glu His His Pro Ile Pro Gln Glu Glu Ala 290 295 300 Ser Leu Ala Glu
Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro 305 310 315 320 Pro
Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser 325 330
335 Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His
340 345 350 Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro
Asn Val 355 360 365 His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp
Leu Ile Arg Asp 370 375 380 Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr
Thr Gly Leu Ser Ala Thr 385 390 395 400 Pro Pro Ala Ser Leu Pro Gly
Ser Leu Ser Asn Val Lys Ala Leu Gln 405 410 415 Lys Ser Pro Gly Pro
Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu 420 425 430 Asp Arg Asn
Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp 435 440 445 Trp
Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser 450 455
460 Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala
465 470 475 480 Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln
Leu Gln Ala 485 490 495 Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr
Arg His Val Asn Ile 500 505 510 Leu Leu Phe Met Gly Tyr Ser Thr Lys
Pro Gln Leu Ala Ile Val Thr 515 520 525 Gln Trp Cys Glu Gly Ser Ser
Leu Tyr His His Leu His Ile Ile Glu 530 535 540 Thr Lys Phe Glu Met
Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala 545 550 555 560 Gln Gly
Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu 565 570 575
Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly 580
585 590 Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His
Gln 595 600 605 Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro
Glu Val Ile 610 615 620 Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln
Ser Asp Val Tyr Ala 625 630 635 640 Phe Gly Ile Val Leu Tyr Glu Leu
Met Thr Gly Gln Leu Pro Tyr Ser 645 650 655 Asn Ile Asn Asn Arg Asp
Gln Ile Ile Phe Met Val Gly Arg Gly Tyr 660 665 670 Leu Ser Pro Asp
Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met 675 680 685 Lys Arg
Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro 690 695 700
Leu Phe Pro Gln Asp Leu Ser Ser His His Arg His Lys Asn Phe Arg 705
710 715 720 Trp Leu Gln Val Tyr Met 725 332331DNABos taurus
33ctcagctgcg ccgggtctca caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc
60cgacgccgcc cgggccgccc gggccgtccc tccccgctgc cccccgtcct ccgcctccgc
120ctccccccgc cctcagcctc ccttccccct ccccgcccag cagcggtcgc
tcgggcccgg 180ctctcggtta taagatggcg gcgctgagtg gcggcggcgg
cggcggcggc ggtggcgcgg 240agcagggcca ggctctgttc aacggggaca
tggagcccga ggccggcgcc gcggcctctt 300cggctgcgga ccccgccatt
cccgaggagg tgtggaatat caaacaaatg attaagttga 360cacaggagca
tatagaggcc ctattggaca aatttggtgg ggagcataat ccaccatcaa
420tatatctgga ggcctatgaa gaatacacca gcaagctaga tgccctccaa
caaagagaac 480aacagttatt ggaatccctg gggaatggaa ctgatttttc
tgtttctagc tctgcatcaa 540cggacaccgt tacatcttct tcctcttcta
gcctttcagt gctgccttca tctctttcag 600tttttcaaaa tcccacagat
gtgtcacgga gcaaccccaa gtcaccacaa aaacctatcg 660ttagagtctt
cctgcccaat aaacagagga cagtggtacc tgcacggtgt ggagtcacag
720tccgggacag cctgaagaag gcactgatga tgagaggtct aatcccagag
tgctgtgctg 780tttacagaat tcaggatggg gagaagaaac caattggctg
ggacactgat atttcctggc 840ttactggaga ggagttgcat gtagaagtgt
tggagaatgt tccacttaca acacacaact 900ttgtacggaa aacttttttc
accttagcat tttgtgactt ctgtagaaag ctgcttttcc 960agggattccg
ctgtcaaaca tgtggttata aatttcacca gcgttgtagt acagaggttc
1020cactgatgtg tgttaattat gaccaactag atttgctgtt tgtctccaag
ttctttgaac 1080accacccaat accacaggag gaggcctcct tagcagagac
tacccttcca tgtggctcat 1140ccccttctgc acccccctcc gattctattg
ggcccccaat tctcaccagt ccatctcctt 1200caaaatccat tccaattcca
cagcctttcc gaccagcaga tgaagatcat cgaaatcagt 1260ttggacaacg
agaccggtcc tcatcagctc caaatgtgca tataaacaca atagaacccg
1320tcaatattga tgacttgatt agagaccaag ggtttcgtag tgatggagga
tcaaccacag 1380gtttatccgc cacaccccct gcctcattac ctggctcact
ctctaatgtg aaagcattgc 1440agaaatctcc aggacctcag cgagaaagaa
agtcctcttc atcctcagaa gacaggaatc 1500gaatgaaaac gcttggtaga
cgggattcaa gtgacgattg ggagattcct gatggacaga 1560tcacagtggg
acaaagaatt ggatcagggt catttgggac agtctacaag ggaaagtggc
1620atggtgatgt ggcagtgaaa atgttgaatg tgacagcacc cacacctcag
cagttacagg 1680ccttcaaaaa tgaagtagga gtactcagga aaacgcgaca
tgtgaatatc ctcctcttca 1740tgggttattc aacaaagcca caactggcta
ttgttaccca gtggtgtgag ggctccagtt 1800tatatcatca tctccacatc
attgagacca aattcgagat gatcaaactt atagatattg 1860cacggcagac
tgcacagggc atggattact tacacgccaa gtcaatcatc cacagagacc
1920tcaagagtaa taatattttt cttcatgaag acctcacagt aaaaataggt
gattttggtc 1980tagccacagt gaaatctcga tggagtgggt cccatcagtt
tgaacagttg tctggatcca 2040ttttgtggat ggcaccagaa gtaatcagaa
tgcaagataa aaacccatat agctttcagt 2100cagatgtata tgcatttggg
attgttctgt atgaattgat gaccggacag ttaccttatt 2160caaatatcaa
caacagggac caggtgcttt gtcctccatg ggagtgtaat aaatgctgtg
2220caagggctta cttcccatga gagaagtgag tgaccaacag aaggataatt
tttatggtgg 2280gacgaggata tctgtctcca gatctcagta aggtacggag
taactgtcca a 233134681PRTBos taurus 34Met Ala Ala Leu Ser Gly Gly
Gly Gly Gly Gly Gly Gly Gly Ala Glu 1 5 10 15 Gln Gly Gln Ala Leu
Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala 20 25 30 Ala Ala Ser
Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn 35 40 45 Ile
Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu 50 55
60 Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala
65 70 75 80 Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg
Glu Gln 85 90 95 Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe
Ser Val Ser Ser 100 105 110 Ser Ala Ser Thr Asp Thr Val Thr Ser Ser
Ser Ser Ser Ser Leu Ser 115 120 125 Val Leu Pro Ser Ser Leu Ser Val
Phe Gln Asn Pro Thr Asp Val Ser 130 135 140 Arg Ser Asn Pro Lys Ser
Pro Gln Lys Pro Ile Val Arg Val Phe Leu 145 150 155 160 Pro Asn Lys
Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val 165 170 175 Arg
Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu 180 185
190 Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly
195 200 205 Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His
Val Glu 210 215 220 Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe
Val Arg Lys Thr 225 230 235 240 Phe Phe Thr Leu Ala Phe Cys Asp Phe
Cys Arg Lys Leu Leu Phe Gln 245 250 255 Gly Phe Arg Cys Gln Thr Cys
Gly Tyr Lys Phe His Gln Arg Cys Ser 260 265 270 Thr Glu Val Pro Leu
Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu 275 280 285 Phe Val Ser
Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala 290 295 300 Ser
Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro 305 310
315 320 Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro
Ser 325 330 335 Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp
Glu Asp His 340 345 350 Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser
Ser Ala Pro Asn Val 355 360 365 His Ile Asn Thr Ile Glu Pro Val Asn
Ile Asp Asp Leu Ile Arg Asp 370 375 380 Gln Gly Phe Arg Ser Asp Gly
Gly Ser Thr Thr Gly Leu Ser Ala Thr 385 390 395 400 Pro Pro Ala Ser
Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln 405 410 415 Lys Ser
Pro Gly Pro Gln Arg Glu Arg
Lys Ser Ser Ser Ser Ser Glu 420 425 430 Asp Arg Asn Arg Met Lys Thr
Leu Gly Arg Arg Asp Ser Ser Asp Asp 435 440 445 Trp Glu Ile Pro Asp
Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser 450 455 460 Gly Ser Phe
Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala 465 470 475 480
Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala 485
490 495 Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn
Ile 500 505 510 Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala
Ile Val Thr 515 520 525 Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His
Leu His Ile Ile Glu 530 535 540 Thr Lys Phe Glu Met Ile Lys Leu Ile
Asp Ile Ala Arg Gln Thr Ala 545 550 555 560 Gln Gly Met Asp Tyr Leu
His Ala Lys Ser Ile Ile His Arg Asp Leu 565 570 575 Lys Ser Asn Asn
Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly 580 585 590 Asp Phe
Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln 595 600 605
Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile 610
615 620 Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr
Ala 625 630 635 640 Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln
Leu Pro Tyr Ser 645 650 655 Asn Ile Asn Asn Arg Asp Gln Val Leu Cys
Pro Pro Trp Glu Cys Asn 660 665 670 Lys Cys Cys Ala Arg Ala Tyr Phe
Pro 675 680 35 2319DNABos taurus 35ctcagctgcg ccgggtctca caagacggtt
cccgaggtgg cccaggcgcc gtcccaccgc 60cgacgccgcc cgggccgccc gggccgtccc
tccccgctgc cccccgtcct ccgcctccgc 120ctccccccgc cctcagcctc
ccttccccct ccccgcccag cagcggtcgc tcgggcccgg 180ctctcggtta
taagatggcg gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg
240agcagggcca ggctctgttc aacggggaca tggagcccga ggccggcgcc
gcggcctctt 300cggctgcgga ccccgccatt cccgaggagg tgtggaatat
caaacaaatg attaagttga 360cacaggagca tatagaggcc ctattggaca
aatttggtgg ggagcataat ccaccatcaa 420tatatctgga ggcctatgaa
gaatacacca gcaagctaga tgccctccaa caaagagaac 480aacagttatt
ggaatccctg gggaatggaa ctgatttttc tgtttctagc tctgcatcaa
540cggacaccgt tacatcttct tcctcttcta gcctttcagt gctgccttca
tctctttcag 600tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa
gtcaccacaa aaacctatcg 660ttagagtctt cctgcccaat aaacagagga
cagtggtacc tgcacggtgt ggagtcacag 720tccgggacag cctgaagaag
gcactgatga tgagaggtct aatcccagag tgctgtgctg 780tttacagaat
tcaggatggg gagaagaaac caattggctg ggacactgat atttcctggc
840ttactggaga ggagttgcat gtagaagtgt tggagaatgt tccacttaca
acacacaact 900ttgtacggaa aacttttttc accttagcat tttgtgactt
ctgtagaaag ctgcttttcc 960agggattccg ctgtcaaaca tgtggttata
aatttcacca gcgttgtagt acagaggttc 1020cactgatgtg tgttaattat
gaccaactag atttgctgtt tgtctccaag ttctttgaac 1080accacccaat
accacaggag gaggcctcct tagcagagac tacccttcca tgtggctcat
1140ccccttctgc acccccctcc gattctattg ggcccccaat tctcaccagt
ccatctcctt 1200caaaatccat tccaattcca cagcctttcc gaccagcaga
tgaagatcat cgaaatcagt 1260ttggacaacg agaccggtcc tcatcagctc
caaatgtgca tataaacaca atagaacccg 1320tcaatattga tgacttgatt
agagaccaag ggtttcgtag tgatggagga tcaaccacag 1380gtttatccgc
cacaccccct gcctcattac ctggctcact ctctaatgtg aaagcattgc
1440agaaatctcc aggacctcag cgagaaagaa agtcctcttc atcctcagaa
gacaggaatc 1500gaatgaaaac gcttggtaga cgggattcaa gtgacgattg
ggagattcct gatggacaga 1560tcacagtggg acaaagaatt ggatcagggt
catttgggac agtctacaag ggaaagtggc 1620atggtgatgt ggcagtgaaa
atgttgaatg tgacagcacc cacacctcag cagttacagg 1680ccttcaaaaa
tgaagtagga gtactcagga aaacgcgaca tgtgaatatc ctcctcttca
1740tgggttattc aacaaagcca caactggcta ttgttaccca gtggtgtgag
ggctccagtt 1800tatatcatca tctccacatc attgagacca aattcgagat
gatcaaactt atagatattg 1860cacggcagac tgcacagggc atggattact
tacacgccaa gtcaatcatc cacagagacc 1920tcaagagtaa taatattttt
cttcatgaag acctcacagt aaaaataggt gattttggtc 1980tagccacagt
gaaatctcga tggagtgggt cccatcagtt tgaacagttg tctggatcca
2040ttttgtggat ggcaccagaa gtaatcagaa tgcaagataa aaacccatat
agctttcagt 2100cagatgtata tgcatttggg attgttctgt atgaattgat
gaccggacag ttaccttatt 2160caaatatcaa caacagggac caggtgcttt
gtcctccatg ggagtgtaat aaatgctgtg 2220caagggctta cttcccatga
gagaagtgag tgaccaacag aaggtctgtg caaggaaaag 2280agacaaagcc
acggatcaga agcacatggc cataactga 231936681PRTBos taurus 36Met Ala
Ala Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu 1 5 10 15
Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala 20
25 30 Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp
Asn 35 40 45 Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu
Ala Leu Leu 50 55 60 Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser
Ile Tyr Leu Glu Ala 65 70 75 80 Tyr Glu Glu Tyr Thr Ser Lys Leu Asp
Ala Leu Gln Gln Arg Glu Gln 85 90 95 Gln Leu Leu Glu Ser Leu Gly
Asn Gly Thr Asp Phe Ser Val Ser Ser 100 105 110 Ser Ala Ser Thr Asp
Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser 115 120 125 Val Leu Pro
Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser 130 135 140 Arg
Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu 145 150
155 160 Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr
Val 165 170 175 Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu
Ile Pro Glu 180 185 190 Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu
Lys Lys Pro Ile Gly 195 200 205 Trp Asp Thr Asp Ile Ser Trp Leu Thr
Gly Glu Glu Leu His Val Glu 210 215 220 Val Leu Glu Asn Val Pro Leu
Thr Thr His Asn Phe Val Arg Lys Thr 225 230 235 240 Phe Phe Thr Leu
Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln 245 250 255 Gly Phe
Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser 260 265 270
Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu 275
280 285 Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu
Ala 290 295 300 Ser Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro
Ser Ala Pro 305 310 315 320 Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu
Thr Ser Pro Ser Pro Ser 325 330 335 Lys Ser Ile Pro Ile Pro Gln Pro
Phe Arg Pro Ala Asp Glu Asp His 340 345 350 Arg Asn Gln Phe Gly Gln
Arg Asp Arg Ser Ser Ser Ala Pro Asn Val 355 360 365 His Ile Asn Thr
Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp 370 375 380 Gln Gly
Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr 385 390 395
400 Pro Pro Ala Ser Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln
405 410 415 Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser
Ser Glu 420 425 430 Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp
Ser Ser Asp Asp 435 440 445 Trp Glu Ile Pro Asp Gly Gln Ile Thr Val
Gly Gln Arg Ile Gly Ser 450 455 460 Gly Ser Phe Gly Thr Val Tyr Lys
Gly Lys Trp His Gly Asp Val Ala 465 470 475 480 Val Lys Met Leu Asn
Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala 485 490 495 Phe Lys Asn
Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile 500 505 510 Leu
Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr 515 520
525 Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu
530 535 540 Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln
Thr Ala 545 550 555 560 Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile
Ile His Arg Asp Leu 565 570 575 Lys Ser Asn Asn Ile Phe Leu His Glu
Asp Leu Thr Val Lys Ile Gly 580 585 590 Asp Phe Gly Leu Ala Thr Val
Lys Ser Arg Trp Ser Gly Ser His Gln 595 600 605 Phe Glu Gln Leu Ser
Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile 610 615 620 Arg Met Gln
Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala 625 630 635 640
Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser 645
650 655 Asn Ile Asn Asn Arg Asp Gln Val Leu Cys Pro Pro Trp Glu Cys
Asn 660 665 670 Lys Cys Cys Ala Arg Ala Tyr Phe Pro 675 680 37
2661DNABos taurus 37tcagctgcgc cgggtctcac aagacggttc ccgaggtggc
ccaggcgccg tcccaccgcc 60gacgccgccc gggccgcccg ggccgtccct ccccgctgcc
ccccgtcctc cgcctccgcc 120tccccccgcc ctcagcctcc cttccccctc
cccgcccagc agcggtcgct cgggcccggc 180tctcggttat aagatggcgg
cgctgagtgg cggcggcggc ggcggcggcg gtggcgcgga 240gcagggccag
gctctgttca acggggacat ggagcccgag gccggcgccg cggcctcttc
300ggctgcggac cccgccattc ccgaggaggt gtggaatatc aaacaaatga
ttaagttgac 360acaggagcat atagaggccc tattggacaa atttggtggg
gagcataatc caccatcaat 420atatctggag gcctatgaag aatacaccag
caagctagat gccctccaac aaagagaaca 480acagttattg gaatccctgg
ggaatggaac tgatttttct gtttctagct ctgcatcaac 540ggacaccgtt
acatcttctt cctcttctag cctttcagtg ctgccttcat ctctttcagt
600ttttcaaaat cccacagatg tgtcacggag caaccccaag tcaccacaaa
aacctatcgt 660tagagtcttc ctgcccaata aacagaggac agtggtacct
gcacggtgtg gagtcacagt 720ccgggacagc ctgaagaagg cactgatgat
gagaggtcta atcccagagt gctgtgctgt 780ttacagaatt caggatgggg
agaagaaacc aattggctgg gacactgata tttcctggct 840tactggagag
gagttgcatg tagaagtgtt ggagaatgtt ccacttacaa cacacaactt
900tgtacggaaa acttttttca ccttagcatt ttgtgacttc tgtagaaagc
tgcttttcca 960gggattccgc tgtcaaacat gtggttataa atttcaccag
cgttgtagta cagaggttcc 1020actgatgtgt gttaattatg accaactaga
tttgctgttt gtctccaagt tctttgaaca 1080ccacccaata ccacaggagg
aggcctcctt agcagagact acccttccat gtggctcatc 1140cccttctgca
cccccctccg attctattgg gcccccaatt ctcaccagtc catctccttc
1200aaaatccatt ccaattccac agcctttccg accagcagat gaagatcatc
gaaatcagtt 1260tggacaacga gaccggtcct catcagctcc aaatgtgcat
ataaacacaa tagaacccgt 1320caatattgat gacttgatta gagaccaagg
gtttcgtagt gatggaggat caaccacagg 1380tttatccgcc acaccccctg
cctcattacc tggctcactc tctaatgtga aagcattgca 1440gaaatctcca
ggacctcagc gagaaagaaa gtcctcttca tcctcagaag acaggaatcg
1500aatgaaaacg cttggtagac gggattcaag tgacgattgg gagattcctg
atggacagat 1560cacagtggga caaagaattg gatcagggtc atttgggaca
gtctacaagg gaaagtggca 1620tggtgatgtg gcagtgaaaa tgttgaatgt
gacagcaccc acacctcagc agttacaggc 1680cttcaaaaat gaagtaggag
tactcaggaa aacgcgacat gtgaatatcc tcctcttcat 1740gggttattca
acaaagccac aactggctat tgttacccag tggtgtgagg gctccagttt
1800atatcatcat ctccacatca ttgagaccaa attcgagatg atcaaactta
tagatattgc 1860acggcagact gcacagggca tggattactt acacgccaag
tcaatcatcc acagagacct 1920caagagtaat aatatttttc ttcatgaaga
cctcacagta aaaataggtg attttggtct 1980agccacagtg aaatctcgat
ggagtgggtc ccatcagttt gaacagttgt ctggatccat 2040tttgtggatg
gcaccagaag taatcagaat gcaagataaa aacccatata gctttcagtc
2100agatgtatat gcatttggga ttgttctgta tgaattgatg accggacagt
taccttattc 2160aaatatcaac aacagggacc agtctgtgca aggaaaagag
acaaagccac ggatcagaag 2220cacatggcca taactgaaga ttttgtgaac
tctcacaagg aaaaaatttg ctctttgaac 2280aataagaagg aactcactaa
aatgtaactg agaactgttc aacaggttga aagctgaaag 2340atgccattgg
aactgacaaa atgtttctta aacataaatg atgaaacagt gaaactacat
2400aatatctcct ctggctgaaa cattcaagaa gtttaaaatg cttaagttaa
aaataaaatc 2460ctagtaaaca atggacttac tgtgcaacat agagaatatc
ttacgataac ctgtaatgga 2520aaagaatctg aaaaagaatg tatataactg
aatcactttg ctgtaaacta gaatctgaca 2580caacactgta aatcactaca
cttttctgtt gcatgccaaa gattatttaa taacgtcatt 2640aaaaaattat
tttaataatt a 266138679PRTBos taurus 38Met Ala Ala Leu Ser Gly Gly
Gly Gly Gly Gly Gly Gly Gly Ala Glu 1 5 10 15 Gln Gly Gln Ala Leu
Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala 20 25 30 Ala Ala Ser
Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn 35 40 45 Ile
Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu 50 55
60 Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala
65 70 75 80 Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg
Glu Gln 85 90 95 Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe
Ser Val Ser Ser 100 105 110 Ser Ala Ser Thr Asp Thr Val Thr Ser Ser
Ser Ser Ser Ser Leu Ser 115 120 125 Val Leu Pro Ser Ser Leu Ser Val
Phe Gln Asn Pro Thr Asp Val Ser 130 135 140 Arg Ser Asn Pro Lys Ser
Pro Gln Lys Pro Ile Val Arg Val Phe Leu 145 150 155 160 Pro Asn Lys
Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val 165 170 175 Arg
Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu 180 185
190 Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly
195 200 205 Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His
Val Glu 210 215 220 Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe
Val Arg Lys Thr 225 230 235 240 Phe Phe Thr Leu Ala Phe Cys Asp Phe
Cys Arg Lys Leu Leu Phe Gln 245 250 255 Gly Phe Arg Cys Gln Thr Cys
Gly Tyr Lys Phe His Gln Arg Cys Ser 260 265 270 Thr Glu Val Pro Leu
Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu 275 280 285 Phe Val Ser
Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala 290 295 300 Ser
Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro 305 310
315 320 Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro
Ser 325 330 335 Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp
Glu Asp His 340 345 350 Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser
Ser Ala Pro Asn Val 355 360 365 His Ile Asn Thr Ile Glu Pro Val Asn
Ile Asp Asp Leu Ile Arg Asp 370 375 380 Gln Gly Phe Arg Ser Asp Gly
Gly Ser Thr Thr Gly Leu Ser Ala Thr 385 390 395 400 Pro Pro Ala Ser
Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln 405 410 415 Lys Ser
Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu 420 425 430
Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp 435
440 445 Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly
Ser 450 455 460 Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly
Asp Val Ala 465 470 475 480 Val Lys Met Leu Asn Val Thr Ala Pro Thr
Pro Gln Gln Leu Gln Ala 485 490 495 Phe Lys Asn Glu Val Gly Val Leu
Arg Lys Thr Arg His Val Asn Ile 500 505 510 Leu Leu Phe Met Gly Tyr
Ser Thr Lys Pro Gln Leu Ala Ile Val Thr 515 520 525 Gln Trp Cys Glu
Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu 530 535 540 Thr Lys
Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala 545 550 555
560 Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu
565 570 575 Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val
Lys
Ile Gly 580 585 590 Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser
Gly Ser His Gln 595 600 605 Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp
Met Ala Pro Glu Val Ile 610 615 620 Arg Met Gln Asp Lys Asn Pro Tyr
Ser Phe Gln Ser Asp Val Tyr Ala 625 630 635 640 Phe Gly Ile Val Leu
Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser 645 650 655 Asn Ile Asn
Asn Arg Asp Gln Ser Val Gln Gly Lys Glu Thr Lys Pro 660 665 670 Arg
Ile Arg Ser Thr Trp Pro 675 39 7434DNABos taurus 39acaccgttac
atcttcttcc tcttctagcc tttcagtgct gccttcatct ctttcagttt 60ttcaaaatcc
cacagatgtg tcacggagca accccaagtc accacaaaaa cctatcgtta
120gagtcttcct gcccaataaa cagaggacag tggtacctgc acggtgtgga
gtcacagtcc 180gggacagcct gaagaaggca ctgatgatga gaggtctaat
cccagagtgc tgtgctgttt 240acagaattca ggatggggag aagaaaccaa
ttggctggga cactgatatt tcctggctta 300ctggagagga gttgcatgta
gaagtgttgg agaatgttcc acttacaaca cacaactttg 360tacggaaaac
ttttttcacc ttagcatttt gtgacttctg tagaaagctg cttttccagg
420gattccgctg tcaaacatgt ggttataaat ttcaccagcg ttgtagtaca
gaggttccac 480tgatgtgtgt taattatgac caactagatt tgctgtttgt
ctccaagttc tttgaacacc 540acccaatacc acaggaggag gcctccttag
cagagactac ccttccatgt ggctcatccc 600cttctgcacc cccctccgat
tctattgggc ccccaattct caccagtcca tctccttcaa 660aatccattcc
aattccacag cctttccgac cagcagatga agatcatcga aatcagtttg
720gacaacgaga ccggtcctca tcagctccaa atgtgcatat aaacacaata
gaacccgtca 780atattgatga cttgattaga gaccaagggt ttcgtagtga
tggaggatca accacaggtt 840tatccgccac accccctgcc tcattacctg
gctcactctc taatgtgaaa gcattgcaga 900aatctccagg acctcagcga
gaaagaaagt cctcttcatc ctcagaagac aggaatcgaa 960tgaaaacgct
tggtagacgg gattcaagtg acgattggga gattcctgat ggacagatca
1020cagtgggaca aagaattgga tcagggtcat ttgggacagt ctacaaggga
aagtggcatg 1080gtgatgtggc agtgaaaatg ttgaatgtga cagcacccac
acctcagcag ttacaggcct 1140tcaaaaatga agtaggagta ctcaggaaaa
cgcgacatgt gaatatcctc ctcttcatgg 1200gttattcaac aaagccacaa
ctggctattg ttacccagtg gtgtgagggc tccagtttat 1260atcatcatct
ccacatcatt gagaccaaat tcgagatgat caaacttata gatattgcac
1320ggcagactgc acagggcatg gattacttac acgccaagtc aatcatccac
agagacctca 1380agagtaataa tatttttctt catgaagacc tcacagtaaa
aataggtgat tttggtctag 1440ccacagtgaa atctcgatgg agtgggtccc
atcagtttga acagttgtct ggatccattt 1500tgtggatggc accagaagta
atcagaatgc aagataaaaa cccatatagc tttcagtcag 1560atgtatatgc
atttgggatt gttctgtatg aattgatgac cggacagtta ccttattcaa
1620atatcaacaa cagggaccag ataattttta tggtgggacg aggatatctg
tctccagatc 1680tcagtaaggt acggagtaac tgtccaaaag ccatgaagag
attaatggca gagtgcctaa 1740aaaagaaaag agatgaaaga ccactctttc
cccaagtagg aaagactctc ctaagcaaga 1800gacaaaattc agaagttatc
agggaaaaag ataagcagat tctcgcctct attgagctgc 1860tggcccgctc
attgccaaaa attcaccgca gtgcatcaga accctccttg aatcgggctg
1920gcttccaaac agaggatttt agtctatatg cttgtgcttc tccaaaaaca
cccattcagg 1980cagggggata tggagaattt gcagccttca agtagccaca
ccatcatgac agcatctact 2040cttatttctt aagtcttgtg ttcgtacaat
ttgttaacat caaaacacag ttctgttcct 2100caactctttt taaagttaaa
atttttcagt gcataagctg gtgtggaaca gaaggaaatt 2160tcccatccaa
caaaagaggg aagaatgttt taggaaccag aattctctgc tgccagtgtt
2220tcttcttcaa cacaaatatc acaagtctgc ccactcccag gaagaaagag
gagagaccct 2280gagttctgac cttttgatgg tcaggcatga tggaaagaaa
ctgctgctac agcttgggag 2340atttgctctg ggaagtctgc cagtcaactt
tgcccttcta accaccagat caatatgtgg 2400ctgatcatct gatggggcag
ttgcaatcac caagccttgt tctctttcct gttctgggat 2460tgtgttgtgg
aacccttttc cctagccacc accagttcat ttctgaggga tggaacaaaa
2520atgcagcatg cccttcctgt gtggtgcatg ttcagtcctt gacaaatttt
taccaaaatg 2580aagctacttt atttaaaagg agggtgagag gtgaggaggt
cactttgggt gtggcggaaa 2640gggaatgctg catctttttc ctgggctgct
ggggctctgg ccttggcttg ccagccggaa 2700gcgctggcac gcatcgcctt
cttttcccat tgggtccagc aatgaagacg agtgtttggg 2760gttttttttt
tctccaccat gtagcaagtt ctcaggaaaa tacaattgat atcttcctcc
2820taagctcttc caatcagtca ccaagtactt atgtggttac tttgtccagg
gcacaaaatg 2880cctgtatcta attaaaagcc tacaaaactg cttgataaca
gttttgaatg tgagacattt 2940atgtaattta aatgtaaggt acaagtttta
atttctgagt ttcttctatt atatttttat 3000taaaaaaaga aaataatttt
cagattgaat tggagtaaaa taatattact tcccactaga 3060attatatatc
ctggaaaatt gtatttttgt tacataagca gcttttaaag aaagatcatt
3120acccttttct ctacataaat atatggggag tcttagccta atgacaaata
tttataattt 3180ttaaattaat ggtacttgct ggatccatac taacatcttt
actaatacct cattgtttct 3240tccaacttac tcctacacta catcctacat
cttcttccta gtcttttatc tagaatatgc 3300aacctcaaat aaaaatggtg
gtgtcctcat tcattctcct ccttcctttt ttcccaagcc 3360tgatcttcaa
aaggttggtt aatttggcag ctgagttcct ccccaggcag agaatagacc
3420aattttaggt gtattgggac tgagggagga tgtgtaaaga ttaacatcag
taaagaaccg 3480ctgtggagta attaagaact ttgttcttta taactggaga
atataaccta accctaacat 3540ccctcagcct ttactaaagt gtggcgtaaa
tcacagtagt agcaaagaaa gtgactctgg 3600atgtgttcct ggccagtacc
tcccttatca tgaatgtaga ctctctcatc aagatttagg 3660aatataaatc
aaatcaaatg tgcccagcca agctatgtag taagggactt gaacaatatt
3720aggcagaacc tataaaataa atcagggaat tagaaattat ttaaagtttt
caaattgtaa 3780attgccccgg tgtctttcag cctactgcca ttatttttgc
tacaatacct acatttcaga 3840ggagggccta ctgaaaattc catgcaagtg
gaaaataatc ctcaagttat taatgagttt 3900gaaaagcaat gagttcttaa
gtctttgtga gtagagcaag atcctacaaa attcagaaat 3960agtaaaaatg
gattcatgct gatttgaaga gcatctgtgt gcataatata atgctgcatc
4020tcttttaaaa gcagtctatt tttcttttta aatttgtccc catagatgct
tttgaacatg 4080aacatgctta tgttaccttt tccgaggttg ggaagagcca
ggagctctca ggcagggccc 4140cctccctcag ctgggcagga gctgctcagg
aggagctagt tatagaggaa gcttagcgtt 4200ggcattttca aaattcaagg
tgataacgct ttcttcttcc tttctgtttt agaatagatt 4260gctgtctgat
ttgaaaaagg gaaatagatt tgatctcaaa tgaatctgtg cccagaagcc
4320aggctcaggg tattcagaga tttgtatagt gccctcaaaa aataacaaaa
ttttagcttt 4380ccttttttct tcttttctcc atcaaattct tttttctcta
gtttacaaat gacatggaaa 4440aggaatttcc cctgagtttt gtatgccttt
ttttttttgg cttagactat agataggcgt 4500gttgagctcc taagaaaata
caaggaggaa ctctttgttg tgcagagcac tttatgagta 4560gtttgtgtgg
ataatatgtg actgcttccc tgacgagctt gtgaggctgt acttatgtct
4620ttcctgtaag gcagcttcag tgccttctgt agtgtatata aggaaagatt
acgccttctg 4680aaaaatctca gagcaaccat aagattattt taaaatatgt
agtatgactg atggactttt 4740tcatcattaa attagtctag catctaaact
tttaccactg aaataatatt gaccaaaaag 4800caatttataa aaggtatttg
tgaatagaaa atacaatgtg atcatttgta cttatgtgca 4860ccttaaaaga
ggaattctgt ctagctgtca aattctggtt ccttaacatc cagtccttga
4920ttgtgattga gatctggtag gacgtgctgg ggcacgctag cagataaaat
cccgtatact 4980ttaggataga tgttacattt atgtcagtgt tggcaaagag
cattgtgtag taataaagaa 5040ttcaagactt cagcaatgtc aacctgaaac
tttgtaaata tttcctagat tgttatttga 5100tgcagtcaca gctctttatc
acacaatgtt gtctttccct catcaggcaa ttttagaact 5160gctgcacacc
cctcctcaga tctcacctgc ccctcctgta cattcacctc tccagccttg
5220tgcacacctc atttagcttt agtttgaaac acattgcagg gttcaggtga
cctcttcaaa 5280aactacctcc tcagaatgag gtaatgaata gttatttatt
ttaaaatatg aaaagtcagg 5340agctctagaa tatgaagatg atctaagatt
ttaactttta tgtatacttg ttgagcactc 5400tccttttgtc ctaaagggca
ttatacattt aagcagtaat actgaaaaat gtagctcaga 5460gtaactgaat
gttgttgaaa gtggtgccag aatctgtttt aggggtacgt atcagaatct
5520taatcttaaa tcggttacat gaaattaaat agttaatggt aacacttgac
taacagatat 5580aattttaatt ttcggtaggc ttttagcaag acagtaagta
catcttcata atgagttagc 5640cacagcttca tcacatgcac agattttcct
gttgagagac tgcccagtta agagggtaga 5700atgatgaacc atttttcagg
attctcttct ttgtccaaac tggcattgtg agtgctagaa 5760tatcagcact
ttcaaactag tgattccaac tattaggcta ttaaaaagca aaacaaacca
5820aacaaaccat agccagacat gggaagttta ctatgagtat aaacagcaaa
tagcttacag 5880gtcatacatt gaaatggtgt aggtaaggcg ttagaaaaat
accttgacaa tttgccaaat 5940gatcttactg tgccttcatg atgcaataaa
aaaaaaaaaa atttagcata aatcagtgat 6000ttgtgaagag agcagccacc
ctggtctaac tcagctgtgt taatattttt tagcgtgcaa 6060tttagactgc
aaagataaat gcactaaaga gtttatagcc aaaatcacat ttaaaaaatg
6120agagaaaaca caggtaaatt ttcagtgaac aaaattattt ttttaaagta
cataatccct 6180agtatagtca gatatattta tcacatagag caaataggtt
gaaatcacaa ttcagtgaca 6240tttctagaga aactttttct actcccatag
gttcttcaaa gcatggaact tttatataac 6300agaaatgtgt gacggtcatt
ttaaattgct gtagtttggg gctgaagtac tgtgtgctgg 6360gcagcaatca
catgtattaa ctagtgagaa aggagaaatt aagatatagg acagaatttg
6420attttcttgt tcccagatta ctgctgccaa cctagacact gagtttccag
aggctgaaac 6480gtaaacttgc agctcagcaa ctgttttgca aagttagtgg
gactgtcctg cttatgctgt 6540tcaaaaatgc tctgagggcc aggtggggcc
tccaggggct cctctctgag gggacatcag 6600actagctaac gacctggcgg
gcggatgtga accggacaca ctccatggtg tgcttcttgt 6660atcggtccct
cgccaccctc aagaaaggct tcagcgggtt ctctagacgt ctccactaag
6720gtgtgttact aacagccatg ggttgttgag cacccgagga gtgcaatagc
atctctgcat 6780gattgtatat tggcccgaag agaatgaagt ggccagtgta
ctcatgttcc atgttgctag 6840ctctggtaaa ctgaaaatac tggtaagatt
tttgttttat cagtacacta gagagtaagc 6900tttgttttgt tgtttttaga
taatgttttc acttccattt ggaaagacat ttaaattgag 6960tttcagtcct
aaattttgcc agtcatggta attagcagtt tctatcaggt atttttaagg
7020tagaagagga tagaaacata agttctaaaa gcttaaggta accgtggttt
attttaaaat 7080gtttaggggt ggttagtctc tacctcaaaa aaagtgagtg
aatcttttat ttcagcattc 7140acaagttcgg ctgttgtttt tgtaatacat
ttttttttta accttttgac ccccctttac 7200ctaagtgtca atgtagtttt
attaattact aagtcagttt cattaaaatg tttatttagc 7260agttttgact
aattgcaatg attaatatag ccagttgtgc atgaggacac agccagtgag
7320tatatctggg ttttttttgt gatgcttttt ttcttaagac ttctgtagat
ttatgaagta 7380ctcattgaaa acaactaaaa tacgtttatt cgtgttaata
tggaaaaaaa aaaa 743440603PRTBos taurus 40Met Met Arg Gly Leu Ile
Pro Glu Cys Cys Ala Val Tyr Arg Ile Gln 1 5 10 15 Asp Gly Glu Lys
Lys Pro Ile Gly Trp Asp Thr Asp Ile Ser Trp Leu 20 25 30 Thr Gly
Glu Glu Leu His Val Glu Val Leu Glu Asn Val Pro Leu Thr 35 40 45
Thr His Asn Phe Val Arg Lys Thr Phe Phe Thr Leu Ala Phe Cys Asp 50
55 60 Phe Cys Arg Lys Leu Leu Phe Gln Gly Phe Arg Cys Gln Thr Cys
Gly 65 70 75 80 Tyr Lys Phe His Gln Arg Cys Ser Thr Glu Val Pro Leu
Met Cys Val 85 90 95 Asn Tyr Asp Gln Leu Asp Leu Leu Phe Val Ser
Lys Phe Phe Glu His 100 105 110 His Pro Ile Pro Gln Glu Glu Ala Ser
Leu Ala Glu Thr Thr Leu Pro 115 120 125 Cys Gly Ser Ser Pro Ser Ala
Pro Pro Ser Asp Ser Ile Gly Pro Pro 130 135 140 Ile Leu Thr Ser Pro
Ser Pro Ser Lys Ser Ile Pro Ile Pro Gln Pro 145 150 155 160 Phe Arg
Pro Ala Asp Glu Asp His Arg Asn Gln Phe Gly Gln Arg Asp 165 170 175
Arg Ser Ser Ser Ala Pro Asn Val His Ile Asn Thr Ile Glu Pro Val 180
185 190 Asn Ile Asp Asp Leu Ile Arg Asp Gln Gly Phe Arg Ser Asp Gly
Gly 195 200 205 Ser Thr Thr Gly Leu Ser Ala Thr Pro Pro Ala Ser Leu
Pro Gly Ser 210 215 220 Leu Ser Asn Val Lys Ala Leu Gln Lys Ser Pro
Gly Pro Gln Arg Glu 225 230 235 240 Arg Lys Ser Ser Ser Ser Ser Glu
Asp Arg Asn Arg Met Lys Thr Leu 245 250 255 Gly Arg Arg Asp Ser Ser
Asp Asp Trp Glu Ile Pro Asp Gly Gln Ile 260 265 270 Thr Val Gly Gln
Arg Ile Gly Ser Gly Ser Phe Gly Thr Val Tyr Lys 275 280 285 Gly Lys
Trp His Gly Asp Val Ala Val Lys Met Leu Asn Val Thr Ala 290 295 300
Pro Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu Val Gly Val Leu 305
310 315 320 Arg Lys Thr Arg His Val Asn Ile Leu Leu Phe Met Gly Tyr
Ser Thr 325 330 335 Lys Pro Gln Leu Ala Ile Val Thr Gln Trp Cys Glu
Gly Ser Ser Leu 340 345 350 Tyr His His Leu His Ile Ile Glu Thr Lys
Phe Glu Met Ile Lys Leu 355 360 365 Ile Asp Ile Ala Arg Gln Thr Ala
Gln Gly Met Asp Tyr Leu His Ala 370 375 380 Lys Ser Ile Ile His Arg
Asp Leu Lys Ser Asn Asn Ile Phe Leu His 385 390 395 400 Glu Asp Leu
Thr Val Lys Ile Gly Asp Phe Gly Leu Ala Thr Val Lys 405 410 415 Ser
Arg Trp Ser Gly Ser His Gln Phe Glu Gln Leu Ser Gly Ser Ile 420 425
430 Leu Trp Met Ala Pro Glu Val Ile Arg Met Gln Asp Lys Asn Pro Tyr
435 440 445 Ser Phe Gln Ser Asp Val Tyr Ala Phe Gly Ile Val Leu Tyr
Glu Leu 450 455 460 Met Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn Asn
Arg Asp Gln Ile 465 470 475 480 Ile Phe Met Val Gly Arg Gly Tyr Leu
Ser Pro Asp Leu Ser Lys Val 485 490 495 Arg Ser Asn Cys Pro Lys Ala
Met Lys Arg Leu Met Ala Glu Cys Leu 500 505 510 Lys Lys Lys Arg Asp
Glu Arg Pro Leu Phe Pro Gln Val Gly Lys Thr 515 520 525 Leu Leu Ser
Lys Arg Gln Asn Ser Glu Val Ile Arg Glu Lys Asp Lys 530 535 540 Gln
Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser Leu Pro Lys Ile 545 550
555 560 His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg Ala Gly Phe Gln
Thr 565 570 575 Glu Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro Lys Thr
Pro Ile Gln 580 585 590 Ala Gly Gly Tyr Gly Glu Phe Ala Ala Phe Lys
595 600 41 2295DNAEquus caballusmisc_feature(79)..(79)n is a, c, g,
or t 41atgaagacgc tgagcggcgg cggcggcggc gcggagcagg gccaggctct
gttcaacggg 60gacatggaac ccggaggcnc cgcgccggcg cccgcggcct cgtcggccgc
ggaccctgcc 120attcccgagg aggtatggaa tatcaaacaa atgattaaat
tgacacagga acatatagag 180gccctattgg acaaatttgg tggggagcat
aatccaccat caatatatct ggaggcctat 240gaagaataca ccagcaagct
agatgccctc caacaaagag aacaacagtt attggaatcc 300ctggggaatg
gaactgattt ttctgtttct agttctgcat caacggacac cgttacatct
360tcttcctctt ctagcctttc agtgctacct tcatctcttt cagtttttca
aaatcccaca 420gatgtgtcac ggagcaaccc taagtcacca caaaaaccta
tcgttagagt cttcctgccc 480aacaaacaga ggacagtggt acctgcaagg
tgtggcgtta cagtccggga cagtctaaag 540aaagcactga tgatgagagg
tctaatccca gagtgctgtg ctgtttacag aattcaggat 600ggagagaaga
aaccaattgg ctgggacact gatatttcct ggctcactgg agaggaattg
660catgtagaag tgttggagaa tgttccactt acaacacaca actttgtacg
gaaaactttt 720ttcaccttag cattttgtga cttttgtcga aagctgcttt
tccagggttt ccgctgtcaa 780acatgtggtt ataaatttca ccagcgttgt
agtacagagg ttccactgat gtgtgttaat 840tatgaccaac ttgatttgct
gtttgtctcc aagttctttg aacaccaccc agtatcacag 900gaggaggcct
ccttagcaga gactgccctt acatctggat catccccttc tgcacccccc
960tccgattcca ttgggcccca aattctcacc agtccatctc cttcaaaatc
cattccaatt 1020ccacagcctt tccgaccagc agatgaagat catcgaaatc
agtttggaca acgagaccgg 1080tcctcatcag ctccaaatgt acatataaac
acaatagaac ctgtcaatat tgatgacttg 1140attagagacc aagggtttcg
tagtgatgga ggatcaacca caggtttatc tgccaccccc 1200cctgcctcat
tacctggctc actcactaat gtgaaggcat tacagaaatc tccaggacct
1260caacgggaaa ggaaatcatc ttcatcctca gaagacagga atcgaatgaa
aactcttggt 1320agacgggatt caagtgacga ttgggagatt cctgatgggc
agatcacagt gggacaaaga 1380attggatctg ggtcatttgg gacagtctac
aagggaaagt ggcatggtga tgtggcagtg 1440aaaatgttga atgtgacagc
acccacacct cagcagttac aggccttcaa aaatgaagta 1500ggagtactca
ggaaaactcg acatgtgaat atcctactct tcatgggcta ttcaacaaag
1560ccacaactgg ctattgttac ccagtggtgt gagggctcca gcttatatca
ccatctccac 1620atcattgaga ccaaatttga gatgatcaaa cttatagata
ttgctcggca aactgcacag 1680ggcatggatt acttacacgc caagtcaatc
atccacagag acctcaagag taataatatt 1740tttcttcatg aagacctcac
agtaaaaata ggtgattttg gtctagccac agtgaaatct 1800cgatggagtg
ggtcccatca gtttgaacag ttgtctggat ccattttgtg gatggcacca
1860gaagtaatca gaatgcaaga taaaaacccg tatagctttc aatcagatgt
atatgccttt 1920gggattgttc tgtatgaatt gatgactgga cagttacctt
attcaaacat caacaacagg 1980gaccagataa tttttatggt gggaagagga
tatctatctc cagatctcag taaggtacgg 2040agtaactgtc caaaagccat
gaagagatta atggcagagt gcctaaaaaa gaaaagagac 2100gagagaccac
tcttccccca aattctcgcc tctattgagc tgctggcccg ctcattgcca
2160aaaattcacc gcagtgcatc agagccctcc ttgaatcggg ctggcttcca
gacagaggat 2220tttagtctat atgcttgtgc ttctccgaaa acacccatcc
aggcaggggg atatggtgcg 2280tttcctgtcc actga 229542764PRTEquus
caballusmisc_feature(27)..(27)Xaa can be any naturally occurring
amino acid 42Met Lys Thr Leu Ser Gly Gly Gly Gly Gly Ala Glu Gln
Gly Gln Ala 1 5 10 15 Leu Phe Asn Gly Asp Met Glu Pro Gly Gly Xaa
Ala Pro Ala Pro Ala 20 25 30 Ala Ser Ser Ala Ala Asp Pro Ala Ile
Pro Glu Glu Val Trp Asn Ile 35 40 45 Lys Gln Met Ile Lys Leu Thr
Gln Glu His Ile Glu Ala Leu Leu Asp 50 55 60 Lys Phe Gly Gly Glu
His Asn Pro Pro Ser Ile Tyr Leu Glu Ala Tyr 65 70 75
80 Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln Gln
85 90 95 Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser
Ser Ser 100 105 110 Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser
Ser Leu Ser Val 115 120 125 Leu Pro Ser Ser Leu Ser Val Phe Gln Asn
Pro Thr Asp Val Ser Arg 130 135 140 Ser Asn Pro Lys Ser Pro Gln Lys
Pro Ile Val Arg Val Phe Leu Pro 145 150 155 160 Asn Lys Gln Arg Thr
Val Val Pro Ala Arg Cys Gly Val Thr Val Arg 165 170 175 Asp Ser Leu
Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu Cys 180 185 190 Cys
Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly Trp 195 200
205 Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu Val
210 215 220 Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys
Thr Phe 225 230 235 240 Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys
Leu Leu Phe Gln Gly 245 250 255 Phe Arg Cys Gln Thr Cys Gly Tyr Lys
Phe His Gln Arg Cys Ser Thr 260 265 270 Glu Val Pro Leu Met Cys Val
Asn Tyr Asp Gln Leu Asp Leu Leu Phe 275 280 285 Val Ser Lys Phe Phe
Glu His His Pro Val Ser Gln Glu Glu Ala Ser 290 295 300 Leu Ala Glu
Thr Ala Leu Thr Ser Gly Ser Ser Pro Ser Ala Pro Pro 305 310 315 320
Ser Asp Ser Ile Gly Pro Gln Ile Leu Thr Ser Pro Ser Pro Ser Lys 325
330 335 Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His
Arg 340 345 350 Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro
Asn Val His 355 360 365 Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp
Leu Ile Arg Asp Gln 370 375 380 Gly Phe Arg Ser Asp Gly Gly Ser Thr
Thr Gly Leu Ser Ala Thr Pro 385 390 395 400 Pro Ala Ser Leu Pro Gly
Ser Leu Thr Asn Val Lys Ala Leu Gln Lys 405 410 415 Ser Pro Gly Pro
Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu Asp 420 425 430 Arg Asn
Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp Trp 435 440 445
Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser Gly 450
455 460 Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala
Val 465 470 475 480 Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln
Leu Gln Ala Phe 485 490 495 Lys Asn Glu Val Gly Val Leu Arg Lys Thr
Arg His Val Asn Ile Leu 500 505 510 Leu Phe Met Gly Tyr Ser Thr Lys
Pro Gln Leu Ala Ile Val Thr Gln 515 520 525 Trp Cys Glu Gly Ser Ser
Leu Tyr His His Leu His Ile Ile Glu Thr 530 535 540 Lys Phe Glu Met
Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala Gln 545 550 555 560 Gly
Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu Lys 565 570
575 Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly Asp
580 585 590 Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His
Gln Phe 595 600 605 Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro
Glu Val Ile Arg 610 615 620 Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln
Ser Asp Val Tyr Ala Phe 625 630 635 640 Gly Ile Val Leu Tyr Glu Leu
Met Thr Gly Gln Leu Pro Tyr Ser Asn 645 650 655 Ile Asn Asn Arg Asp
Gln Ile Ile Phe Met Val Gly Arg Gly Tyr Leu 660 665 670 Ser Pro Asp
Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met Lys 675 680 685 Arg
Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro Leu 690 695
700 Phe Pro Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser Leu Pro
705 710 715 720 Lys Ile His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg
Ala Gly Phe 725 730 735 Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys Ala
Ser Pro Lys Thr Pro 740 745 750 Ile Gln Ala Gly Gly Tyr Gly Ala Phe
Pro Val His 755 760 43 2678DNAGallus gallus 43tccccctccc tcgccccagc
gcttcgatcc aagatggcgg cgctgagcag cggcagcagc 60gccgaggggg cctcgctctt
caacggggac atggagcccg agccgccgcc gcccgtgctg 120ggcgcctgct
acgccgggag cggcggcggc gacccggcca tcccggagga ggtgtggaat
180atcaaacaga tgattaaatt aacacaagaa catatagaag cgctgttaga
caagtttgga 240ggagagcata acccaccatc aatatattta gaggcctatg
aggagtacac cagcaaacta 300gatgctctac agcagagaga acagcagtta
ttggaatcca tgggaaatgg aactgatttc 360tctgtttcca gttcagcttc
aacggacaca gttgcatcat cttcctcctc tagcctctct 420gtagcacctt
catccctttc agtttatcaa aatcctactg atatgtcgcg gaataaccct
480aagtctccac agaagcctat tgttagagtc ttcctgccca acaagcaaag
gactgtggtt 540ccggcaagat gtggggtgac agtccgagac agcctgaaga
aagctctgat gatgagaggt 600cttattccag aatgctgtgc tgtttacaga
atacaggatg gagagaagaa gccaattggc 660tgggacactg acatttcctg
gctaaccgga gaggagttac acgtggaggt cttggagaat 720gtgccactca
caacacacaa ttttgtacga aaaacattct tcacgttagc gttctgcgac
780ttctgtcgaa agctgctttt ccagggattc cgatgccaga catgtggcta
caaatttcac 840cagcgctgta gcacagaagt gccactgatg tgtgttaact
acgaccaact cgatttgctg 900tttgtctcca agttctttga acatcacccc
atatcgcagg aggagaccac cttaggagag 960accaccccgg catcgggatc
gtacccctca gtgcccccat cagattctgt tggaccacca 1020attctcccta
gtccttctcc ttcaaaatcc attccaatcc cacagccctt ccgaccagca
1080gatgaagacc atcggaatca gtttgggcaa cgcgaccgat cctcttcagc
tcccaatgtt 1140cacatcaata caattgagcc agtcaatatt gatgacttga
ttagagacca gggtgtacga 1200ggagagggag cccctttgaa ccagctgatg
cgctgtcttc ggaaatacca atcccggact 1260cccagtcccc tccttcattc
tgtccccagt gaaatagtgt ttgattttga gcctggccca 1320gtgttcagag
gttcaactgc aggtttgtct gcaacacctc ctgcatcttt gcctgggtca
1380cttaccaatg tgaaagcatt acagaaatca ccaggccccc aacgggaaag
gaaatcatcc 1440tcatcctcag aagacagaaa taggatgaaa acccttggtc
gacgagattc aagtgatgat 1500tgggaaatac cagatgggca gatcacagtt
ggacaaagga taggatctgg atcatttgga 1560acagtctaca aaggaaagtg
gcatggtgac gtggcagtga aaatgttgaa tgttacagca 1620cccacacctc
aacagttaca ggctttcaaa aatgaagtag gagtgctcag gaaaacacgg
1680catgtgaata tcctactttt tatgggttat tcaacaaaac ctcagttggc
tattgttaca 1740cagtggtgtg aggggtccag cttatatcac catctgcaca
taattgagac caagtttgaa 1800atgatcaaac taattgatat tgcacgacag
actgcacaag gcatggatta tttgcatgcc 1860aagtcaatca tccacagaga
cctcaagagt aataatattt ttcttcatga agacctcaca 1920gtaaaaatag
gtgacttcgg tctggctaca gtgaaatcac gatggagtgg atctcatcaa
1980tttgaacagt tatctggatc aattctatgg atggcaccgg aagtgatcag
gatgcaagac 2040aaaaacccat atagctttca gtcagatgtg tatgcattcg
ggattgtgct ttatgaactg 2100atgactggac agttaccata ctcaaacatc
aacaacaggg accagataat ttttatggtg 2160ggacgaggat atctatctcc
agacctcagt aaagtaagaa gtaactgtcc aaaagctatg 2220aagagactaa
tggcagaatg cttgaaaaag aaaagagatg agagacctct ttttccacag
2280attcttgcct ccattgagct tctggcccgg tcgttgccaa aaattcaccg
cagtgcatct 2340gagccgtcac taaaccgggc tggcttccag accgaggatt
tcagtctgta tgcttgtgct 2400tctccaaaaa cgcccatcca agcaggggga
tacggtgggt ttccagtaca ctgaaaagaa 2460atgtgaaagc gtgtgcctgt
ttgctcatgt gctggtgtgt tcctgtgtgt gcaacgcata 2520cgtacgttct
cagttcctac cagcgacttt ttaaggttta ctgagggaat gaagactcat
2580ttcctaacat ggggcattga acgtcctgag cacaagtcag tgctggtaag
gaatgtcttg 2640ggaacagctg gcaagaagaa ttagaaggta cttaaagg
267844806PRTGallus gallus 44Met Ala Ala Leu Ser Ser Gly Ser Ser Ala
Glu Gly Ala Ser Leu Phe 1 5 10 15 Asn Gly Asp Met Glu Pro Glu Pro
Pro Pro Pro Val Leu Gly Ala Cys 20 25 30 Tyr Ala Gly Ser Gly Gly
Gly Asp Pro Ala Ile Pro Glu Glu Val Trp 35 40 45 Asn Ile Lys Gln
Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu 50 55 60 Leu Asp
Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu 65 70 75 80
Ala Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu 85
90 95 Gln Gln Leu Leu Glu Ser Met Gly Asn Gly Thr Asp Phe Ser Val
Ser 100 105 110 Ser Ser Ala Ser Thr Asp Thr Val Ala Ser Ser Ser Ser
Ser Ser Leu 115 120 125 Ser Val Ala Pro Ser Ser Leu Ser Val Tyr Gln
Asn Pro Thr Asp Met 130 135 140 Ser Arg Asn Asn Pro Lys Ser Pro Gln
Lys Pro Ile Val Arg Val Phe 145 150 155 160 Leu Pro Asn Lys Gln Arg
Thr Val Val Pro Ala Arg Cys Gly Val Thr 165 170 175 Val Arg Asp Ser
Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro 180 185 190 Glu Cys
Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile 195 200 205
Gly Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val 210
215 220 Glu Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg
Lys 225 230 235 240 Thr Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg
Lys Leu Leu Phe 245 250 255 Gln Gly Phe Arg Cys Gln Thr Cys Gly Tyr
Lys Phe His Gln Arg Cys 260 265 270 Ser Thr Glu Val Pro Leu Met Cys
Val Asn Tyr Asp Gln Leu Asp Leu 275 280 285 Leu Phe Val Ser Lys Phe
Phe Glu His His Pro Ile Ser Gln Glu Glu 290 295 300 Thr Thr Leu Gly
Glu Thr Thr Pro Ala Ser Gly Ser Tyr Pro Ser Val 305 310 315 320 Pro
Pro Ser Asp Ser Val Gly Pro Pro Ile Leu Pro Ser Pro Ser Pro 325 330
335 Ser Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp
340 345 350 His Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala
Pro Asn 355 360 365 Val His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp
Asp Leu Ile Arg 370 375 380 Asp Gln Gly Val Arg Gly Glu Gly Ala Pro
Leu Asn Gln Leu Met Arg 385 390 395 400 Cys Leu Arg Lys Tyr Gln Ser
Arg Thr Pro Ser Pro Leu Leu His Ser 405 410 415 Val Pro Ser Glu Ile
Val Phe Asp Phe Glu Pro Gly Pro Val Phe Arg 420 425 430 Gly Ser Thr
Ala Gly Leu Ser Ala Thr Pro Pro Ala Ser Leu Pro Gly 435 440 445 Ser
Leu Thr Asn Val Lys Ala Leu Gln Lys Ser Pro Gly Pro Gln Arg 450 455
460 Glu Arg Lys Ser Ser Ser Ser Ser Glu Asp Arg Asn Arg Met Lys Thr
465 470 475 480 Leu Gly Arg Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro
Asp Gly Gln 485 490 495 Ile Thr Val Gly Gln Arg Ile Gly Ser Gly Ser
Phe Gly Thr Val Tyr 500 505 510 Lys Gly Lys Trp His Gly Asp Val Ala
Val Lys Met Leu Asn Val Thr 515 520 525 Ala Pro Thr Pro Gln Gln Leu
Gln Ala Phe Lys Asn Glu Val Gly Val 530 535 540 Leu Arg Lys Thr Arg
His Val Asn Ile Leu Leu Phe Met Gly Tyr Ser 545 550 555 560 Thr Lys
Pro Gln Leu Ala Ile Val Thr Gln Trp Cys Glu Gly Ser Ser 565 570 575
Leu Tyr His His Leu His Ile Ile Glu Thr Lys Phe Glu Met Ile Lys 580
585 590 Leu Ile Asp Ile Ala Arg Gln Thr Ala Gln Gly Met Asp Tyr Leu
His 595 600 605 Ala Lys Ser Ile Ile His Arg Asp Leu Lys Ser Asn Asn
Ile Phe Leu 610 615 620 His Glu Asp Leu Thr Val Lys Ile Gly Asp Phe
Gly Leu Ala Thr Val 625 630 635 640 Lys Ser Arg Trp Ser Gly Ser His
Gln Phe Glu Gln Leu Ser Gly Ser 645 650 655 Ile Leu Trp Met Ala Pro
Glu Val Ile Arg Met Gln Asp Lys Asn Pro 660 665 670 Tyr Ser Phe Gln
Ser Asp Val Tyr Ala Phe Gly Ile Val Leu Tyr Glu 675 680 685 Leu Met
Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn Asn Arg Asp Gln 690 695 700
Ile Ile Phe Met Val Gly Arg Gly Tyr Leu Ser Pro Asp Leu Ser Lys 705
710 715 720 Val Arg Ser Asn Cys Pro Lys Ala Met Lys Arg Leu Met Ala
Glu Cys 725 730 735 Leu Lys Lys Lys Arg Asp Glu Arg Pro Leu Phe Pro
Gln Ile Leu Ala 740 745 750 Ser Ile Glu Leu Leu Ala Arg Ser Leu Pro
Lys Ile His Arg Ser Ala 755 760 765 Ser Glu Pro Ser Leu Asn Arg Ala
Gly Phe Gln Thr Glu Asp Phe Ser 770 775 780 Leu Tyr Ala Cys Ala Ser
Pro Lys Thr Pro Ile Gln Ala Gly Gly Tyr 785 790 795 800 Gly Gly Phe
Pro Val His 805 454454DNAHomo sapiens 45gaaacgtccc gtgtgggagg
ggcgggtctg ggtgcggcct gccgcatgac tcgtggttcg 60gaggcccacg tggccggggc
ggggactcag gcgcctgggg cgccgactga ttacgtagcg 120ggcggggccg
gaagtgccgc tccttggtgg gggctgttca tggcggttcc ggggtctcca
180acatttttcc cggctgtggt cctaaatctg tccaaagcag aggcagtgga
gcttgaggtt 240cttgctggtg tgaaatgact gagtacaaac tggtggtggt
tggagcaggt ggtgttggga 300aaagcgcact gacaatccag ctaatccaga
accactttgt agatgaatat gatcccacca 360tagaggattc ttacagaaaa
caagtggtta tagatggtga aacctgtttg ttggacatac 420tggatacagc
tggacaagaa gagtacagtg ccatgagaga ccaatacatg aggacaggcg
480aaggcttcct ctgtgtattt gccatcaata atagcaagtc atttgcggat
attaacctct 540acagggagca gattaagcga gtaaaagact cggatgatgt
acctatggtg ctagtgggaa 600acaagtgtga tttgccaaca aggacagttg
atacaaaaca agcccacgaa ctggccaaga 660gttacgggat tccattcatt
gaaacctcag ccaagaccag acagggtgtt gaagatgctt 720tttacacact
ggtaagagaa atacgccagt accgaatgaa aaaactcaac agcagtgatg
780atgggactca gggttgtatg ggattgccat gtgtggtgat gtaacaagat
acttttaaag 840ttttgtcaga aaagagccac tttcaagctg cactgacacc
ctggtcctga cttccctgga 900ggagaagtat tcctgttgct gtcttcagtc
tcacagagaa gctcctgcta cttccccagc 960tctcagtagt ttagtacaat
aatctctatt tgagaagttc tcagaataac tacctcctca 1020cttggctgtc
tgaccagaga atgcacctct tgttactccc tgttattttt ctgccctggg
1080ttcttccaca gcacaaacac acctctgcca ccccaggttt ttcatctgaa
aagcagttca 1140tgtctgaaac agagaaccaa accgcaaacg tgaaattcta
ttgaaaacag tgtcttgagc 1200tctaaagtag caactgctgg tgattttttt
tttcttttta ctgttgaact tagaactatg 1260ctaatttttg gagaaatgtc
ataaattact gttttgccaa gaatatagtt attattgctg 1320tttggtttgt
ttataatgtt atcggctcta ttctctaaac tggcatctgc tctagattca
1380taaatacaaa aatgaatact gaattttgag tctatcctag tcttcacaac
tttgacgtaa 1440ttaaatccaa ctttcacagt gaagtgcctt tttcctagaa
gtggtttgta gacttccttt 1500ataatatttc agtggaatag atgtctcaaa
aatccttatg catgaaatga atgtctgaga 1560tacgtctgtg acttatctac
cattgaagga aagctatatc tatttgagag cagatgccat 1620tttgtacatg
tatgaaattg gttttccaga ggcctgtttt ggggctttcc caggagaaag
1680atgaaactga aagcacatga ataatttcac ttaataattt ttacctaatc
tccacttttt 1740tcataggtta ctacctatac aatgtatgta atttgtttcc
cctagcttac tgataaacct 1800aatattcaat gaacttccat ttgtattcaa
atttgtgtca taccagaaag ctctacattt 1860gcagatgttc aaatattgta
aaactttggt gcattgttat ttaatagctg tgatcagtga 1920ttttcaaacc
tcaaatatag tatattaaca aattacattt tcactgtata tcatggtatc
1980ttaatgatgt atataattgc cttcaatccc cttctcaccc caccctctac
agcttccccc 2040acagcaatag gggcttgatt atttcagttg agtaaagcat
ggtgctaatg gaccagggtc 2100acagtttcaa aacttgaaca atccagttag
catcacagag aaagaaattc ttctgcattt 2160gctcattgca ccagtaactc
cagctagtaa ttttgctagg tagctgcagt tagccctgca 2220aggaaagaag
aggtcagtta gcacaaaccc tttaccatga ctggaaaact cagtatcacg
2280tatttaaaca tttttttttc ttttagccat gtagaaactc taaattaagc
caatattctc 2340atttgagaat gaggatgtct cagctgagaa acgttttaaa
ttctctttat tcataatgtt 2400ctttgaaggg tttaaaacaa gatgttgata
aatctaagct gatgagtttg ctcaaaacag 2460gaagttgaaa ttgttgagac
aggaatggaa aatataatta attgatacct atgaggattt 2520ggaggcttgg
cattttaatt tgcagataat accctggtaa ttctcatgaa aaatagactt
2580ggataacttt tgataaaaga ctaattccaa aatggccact ttgttcctgt
ctttaatatc 2640taaatactta ctgaggtcct ccatcttcta tattatgaat
tttcatttat taagcaaatg 2700tcatattacc ttgaaattca gaagagaaga
aacatatact gtgtccagag tataatgaac 2760ctgcagagtt gtgcttctta
ctgctaattc tgggagcttt cacagtactg tcatcatttg 2820taaatggaaa
ttctgctttt ctgtttctgc tccttctgga gcagtgctac tctgtaattt
2880tcctgaggct tatcacctca gtcatttctt ttttaaatgt ctgtgactgg
cagtgattct 2940ttttcttaaa aatctattaa atttgatgtc aaattaggga
gaaagatagt tactcatctt 3000gggctcttgt gccaatagcc cttgtatgta
tgtacttaga gttttccaag tatgttctaa 3060gcacagaagt ttctaaatgg
ggccaaaatt cagacttgag tatgttcttt gaatacctta 3120agaagttaca
attagccggg catggtggcc cgtgcctgta gtcccagcta cttgagaggc
3180tgaggcagga gaatcacttc aacccaggag gtggaggtta cagtgagcag
agatcgtgcc 3240actgcactcc agcctgggtg acaagagaga cttgtctcca
aaaaaaaagt tacacctagg 3300tgtgaatttt ggcacaaagg agtgacaaac
ttatagttaa aagctgaata acttcagtgt 3360ggtataaaac gtggttttta
ggctatgttt gtgattgctg aaaagaattc tagtttacct 3420caaaatcctt
ctctttcccc aaattaagtg cctggccagc tgtcataaat tacatattcc
3480ttttggtttt tttaaaggtt acatgttcaa gagtgaaaat aagatgttct
gtctgaaggc 3540taccatgccg gatctgtaaa tgaacctgtt aaatgctgta
tttgctccaa cggcttacta 3600tagaatgtta cttaatacaa tatcatactt
attacaattt ttactatagg agtgtaatag 3660gtaaaattaa tctctatttt
agtgggccca tgtttagtct ttcaccatcc tttaaactgc 3720tgtgaatttt
tttgtcatga cttgaaagca aggatagaga aacactttag agatatgtgg
3780ggttttttta ccattccaga gcttgtgagc ataatcatat ttgctttata
tttatagtca 3840tgaactccta agttggcagc tacaaccaag aaccaaaaaa
tggtgcgttc tgcttcttgt 3900aattcatctc tgctaataaa ttataagaag
caaggaaaat tagggaaaat attttatttg 3960gatggtttct ataaacaagg
gactataatt cttgtacatt atttttcatc tttgctgttt 4020ctttgagcag
tctaatgtgc cacacaatta tctaaggtat ttgttttcta taagaattgt
4080tttaaaagta ttcttgttac cagagtagtt gtattatatt tcaaaacgta
agatgatttt 4140taaaagcctg agtactgacc taagatggaa ttgtatgaac
tctgctctgg agggagggga 4200ggatgtccgt ggaagttgta agacttttat
ttttttgtgc catcaaatat aggtaaaaat 4260aattgtgcaa ttctgctgtt
taaacaggaa ctattggcct ccttggccct aaatggaagg 4320gccgatattt
taagttgatt attttattgt aaattaatcc aacctagttc tttttaattt
4380ggttgaatgt tttttcttgt taaatgatgt ttaaaaaata aaaactggaa
gttcttggct 4440tagtcataat tctt 445446189PRTHomo sapiens 46Met Thr
Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys 1 5 10 15
Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr 20
25 30 Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp
Gly 35 40 45 Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln
Glu Glu Tyr 50 55 60 Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly
Glu Gly Phe Leu Cys 65 70 75 80 Val Phe Ala Ile Asn Asn Ser Lys Ser
Phe Ala Asp Ile Asn Leu Tyr 85 90 95 Arg Glu Gln Ile Lys Arg Val
Lys Asp Ser Asp Asp Val Pro Met Val 100 105 110 Leu Val Gly Asn Lys
Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys 115 120 125 Gln Ala His
Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr 130 135 140 Ser
Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val 145 150
155 160 Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Ser Asp
Asp 165 170 175 Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val Val Met
180 185 471326DNARattus norvegicus 47gccgttcatg gcggtttcgg
ggtctccaac agcttctcag gttgaaatcc aaaagcctcc 60cgaggcgggg tctgcggagt
ttgagatttt tgcaggtgtg aaatgactga gtacaaactg 120gtggtggttg
gagcaggtgg cgttgggaaa agtgctttga caatccagct aatccagaac
180cactttgtgg atgaatatga tcccaccata gaggattctt accgaaaaca
agtggtgatt 240gacggtgaga cctgtctact ggacatactg gacacagctg
gacaagagga gtacagtgcc 300atgagagacc aatacatgag gacaggcgaa
gggttcctct gtgtgtttgc catcaataat 360agcaaatcct ttgcagatat
taacctctac agggagcaaa ttaagcgcgt gaaagactct 420gatgatgtac
ccatggtgct ggtagggaac aagtgtgact tgccaacaag gacagttgac
480acaaagcaag cccacgagct ggccaagagt tatggaattc cattcattga
aacctcagcc 540aagacccgac agggtgtgga ggatgccttt tacacgcttg
taagggagat acgccagtac 600cggatgaaga agctcaacag cagtgaggat
ggcactcaag gctgtatggg gctgccctgt 660gtggtgatgt agtaagaccc
tttaaaagtt ctgtcatcag aaacgagcca ctttcaagcc 720tcactgatgc
cctggttctg acatccctgg aggagacgtg tttctgctgc tctctgcatc
780tcagagaagc tcctgcttcc tgcttcccca acttagttac tgagcacagc
catctaacct 840gagacctctt cagaataact acctcctcac tcggctgtcc
gaccagagaa atgaacctgt 900ttctccccag tagttctctg ccctgggttt
cccctagaaa caaacacacc tgccagctgg 960ctttgtcctc cgaaaagcag
tttacattga tgcagagaac caaactatag acaagcaatt 1020ctgttgtcaa
cagtttctta agctctaagg taacaattgc tggtgatttc cccctttgcc
1080cccaactgtt gaacttggcc ttgttagttt tgggggaaat gtcaaaaatt
aatctcttcc 1140cgagaataga attagtgttg ctgattgcct gatttgcaat
gtgatcagct atattctata 1200agctggcgtc tgctctgtat tcataaatgc
aaacatgagt actgacgtaa gtgcatccct 1260agtcttctca gctgcatgca
attaaatcca acgttcacaa caaaaaaaaa aaaaaaaaaa 1320aaaaaa
132648189PRTRattus norvegicus 48Met Thr Glu Tyr Lys Leu Val Val Val
Gly Ala Gly Gly Val Gly Lys 1 5 10 15 Ser Ala Leu Thr Ile Gln Leu
Ile Gln Asn His Phe Val Asp Glu Tyr 20 25 30 Asp Pro Thr Ile Glu
Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly 35 40 45 Glu Thr Cys
Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr 50 55 60 Ser
Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys 65 70
75 80 Val Phe Ala Ile Asn Asn Ser Lys Ser Phe Ala Asp Ile Asn Leu
Tyr 85 90 95 Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Asp Asp Val
Pro Met Val 100 105 110 Leu Val Gly Asn Lys Cys Asp Leu Pro Thr Arg
Thr Val Asp Thr Lys 115 120 125 Gln Ala His Glu Leu Ala Lys Ser Tyr
Gly Ile Pro Phe Ile Glu Thr 130 135 140 Ser Ala Lys Thr Arg Gln Gly
Val Glu Asp Ala Phe Tyr Thr Leu Val 145 150 155 160 Arg Glu Ile Arg
Gln Tyr Arg Met Lys Lys Leu Asn Ser Ser Glu Asp 165 170 175 Gly Thr
Gln Gly Cys Met Gly Leu Pro Cys Val Val Met 180 185 49 4470DNAMus
musculus 49gggactgggg cgccttgggc gcctagtgat tacgtagcgg gtggggccgg
aagtgccgct 60ccctggcggg ggctgttcat ggcggtttcg gggtctccaa cagcttctca
ggttgaagtc 120caaaagcctc ccgaggcggg gtctgcggag tttgaggttt
ttgctggtgt gaaatgactg 180agtacaaact ggtggtggtt ggagcaggtg
gtgttgggaa aagcgccttg acgatccagc 240taatccagaa ccactttgtg
gatgaatatg atcccaccat agaggattct taccgaaagc 300aagtggtgat
tgatggtgag acctgcctgc tggacatact ggacacagct ggacaagagg
360agtacagtgc catgagagac cagtacatga ggacaggcga agggttcctc
tgtgtatttg 420ccatcaataa tagcaaatca tttgcagata ttaacctcta
cagggagcaa attaagcgtg 480tgaaagattc tgatgatgtc cccatggtgc
tggtaggcaa caagtgtgac ttgccaacaa 540ggacagttga cacaaagcaa
gcccacgaac tggccaagag ttacggaatt ccattcattg 600agacctcagc
caagacccga cagggtgtgg aggatgcctt ttacacactg gtaagggaga
660tacgccagta ccgaatgaaa aagctcaaca gcagtgacga tggcactcaa
ggttgtatgg 720ggctgccctg tgtgctgatg tagtaagaca ctttgaaagt
tctgtcatca gaaaagagcc 780actttgaagc tgcactgatg ccctggttct
gacatccctg gaggagacct gttcctgctg 840ctctctgcat ctcagagaag
ctcctgcttc ctgcttcccc gactcagtta ctgagcacag 900ccatctaacc
tgagacctct tcagaataac tacctcctca ctcggctgtc tgaccagaga
960aatagacctg tctctcccgg tcgttctctg ccctgggttc ccctagaaac
agacacagcc 1020tccagctggc tttgtcctct gaaaagcagt ttacattgat
gcagagaacc aaactagaca 1080tgccattctg ttgacaacag tttcttatac
tctaaggtaa caactgctgg tgattttccc 1140ctgcccccaa ctgttgaact
tggccttgtt ggtttggggg gaaaatgtca taaattactt 1200tcttcccaaa
atataattag tgttgctgat tgatttgtaa tgtgatcagc tatattccat
1260aaactggcat ctgctctgta ttcataaatg caaacacgaa tactctcaac
tgcatgcaat 1320taaatccaac attcacaaca aagtgccttt ttcctaaaag
tgctctgtag gctccattac 1380agtttgtaat tggaatagat gtgtcaagaa
ccattgtata ggaaagtgac tctgagccat 1440ctacctttga gggaaaggtg
tatgtacctg atggcagatg ctttgtgtat gcacatgaag 1500atagtttccc
tgtctgggat tctcccagga gaaagatgga actgaaacaa ttacaagtaa
1560tttcatttaa ttctagctaa tctttttttt tttttttttt ttttttggta
gactatcacc 1620tataaatatt tggaatatct tctagcttac tgataatcta
ataattaatg agcttccatt 1680ataatgaatt ggttcatacc aggaagccct
ccatttatag tatagatact gtaaaaattg 1740gcatgttgtt actttatagc
tgtgattaat gattcctcag accttgctga gatatagtta 1800ttagcagaca
ggttatatct ttgctgcata gtttcttcat ggaatatata tctatctgta
1860tgtggagaga acgtggccct cagttccctt ctcagcatcc ctcatctctc
agcctagaga 1920agttcgagca tcctagaggg gcttgaacag ttatctcggt
taaaccatgg tgctaatgga 1980ccgggtcatg gtttcaaaac ttgaacaagc
cagttagcat cacagagaaa cagtccatcc 2040atatttgctc cctgcctatt
attcctgctt acagactttt gcctgatgcc tgctgttagt 2100gctacaagga
taaagcttgt gtggttctca ccaggactgg aagtacctgg tgagctctgg
2160ggtaagccta gatatcttta cattttcaga cccttattct tagccacgtg
gaaactgaag 2220ccagagtcca tacctccatc tccttccccc cccaaaaaaa
ttagattaat gttctttata 2280tagctttttt aaagtattta aaacatgtct
ataagttagg ctgccaacta acaaaagctg 2340atgtgtttgt tcaaataaag
aggtatcctt cgctactcga gagaagaatg taaaatgcca 2400ttgattgttg
tcacttggag gcttgatgtt tgccctgata attcattagt gggttttgtt
2460tgtcacatga tacctaagat gtaactcagc tcagtaattc taatgaaaac
ataaattgga 2520taccttaatt gaaaaaagca aacctaattc caaaatggcc
attttctctt ctgatcttgt 2580aatacctaaa attctgaggt ccttgggatt
cttttgttta taacaggatc ttgctgtgta 2640gtcctagctg gcctcaaact
cacaatactc ttcctggatc aatctcccaa gtgctgggat 2700tacaggcaca
ttccaccaca cacacctgac tgagctcgtt cctaatgagt tttcattaag
2760caaattcccc atcaccttga aactaatcag aagggggaag aaacatttgc
tatgctcctg 2820agtgctaaca ctgggatcat tcacatgggg tttgcattcc
taggcaaact aaactgctgc 2880cttttacaac aaggctcagt catcttcctg
aagctgctga gaccagcact tggtcttgtt 2940ttgttttaat atgtctatat
gactggtggt ggatccctaa atagtttatt aattaaactc 3000cagttaagga
gaaagttact caccttgacc cgtttgacca tatcccgtgt gtgtgtgtgt
3060gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgcacgcgt atgtacgtac
gtatgtatgt 3120aggtatgtag gtggtttcca gtataaacac agaaacaaat
ggagccaatt caggtttcag 3180atgcccttac taacatatat tcccacgggg
tgtgggtttt ggcacaacag tgacaaactt 3240aaaagccaag taagagccgg
gcgtggtggc gcacgccttt aatcccagca cttgggaggc 3300agaggcaggc
ggatttctga gttctaggcc atcctggtct acagtgagtt ccaggacagc
3360cagtgctaca cagagaaacc ctgtctcgaa aagccaaaaa aaaaaaaaaa
aaaaaaaaaa 3420aaaagccaag taggtccagt tggtatagta tcaaagtgtt
tttagagtaa ttagtgaagg 3480tctgctttac ctcaaagttg cagagcctct
cttcctgagt ttaagtgcct ggccggcagt 3540cacaaattaa catgttgctg
taaggcagtt agttgaagct ttgttcacac attggagagt 3600atgaaaataa
agtgttctaa gagcgctgat actggatctg tgtaaacctg gtaaatgccg
3660tttgtccagg acttagcgtg tgtgagttgg tagctcagta cgagtttact
agttccgcag 3720tgtgtacaat ggaggcgggt ttgttttagc tggccacctg
tagaatcagc ctttaaactg 3780ctgtgaactt tgtcatgact tgaatatgaa
gatagacaaa aactctgtaa agacaaatgt 3840ttgttttccc ccttacagaa
cgtgtgagct tggttttatc ttcctttgta tttagtcata 3900acctctcaag
ctggcagctc cgaccaagga tcagaagctg tgtgcgttcc acctggtgga
3960attagctcag ctctatatga gaagtggagt taatggaaaa cgtgttgact
gggtggtttc 4020tatttaaaag agtgatgata attcttgaac agtagttttt
attttgctat ttctttaagc 4080tgactgatgt gccacaaaat tattttaagg
tatttgtgtt ttaagagtgt tctcatgaga 4140ttagttgtag atatttttta
aaatacaact ggtttttaaa atctgagtat tgctctaagc 4200aagtgtttag
actcttacgg gaaggtgggt ggaagttgtt tggcttccgt atttccatgc
4260gtgccgtcag acataggtca gaacgccaac tgtgcatcct gctgtttaaa
gacctcttgg 4320cctctgtgac cctcatgaag gggctgatat tttaagttga
ctgtttgatt gtaaattaat 4380cctttctaat ttttaaagac ttgcttgact
gttttccttg ttaaataatt ttaaaaaaat 4440aaaaaactgg aagttctttg
cttaactgta 447050189PRTMus musculus 50Met Thr Glu Tyr Lys Leu Val
Val Val Gly Ala Gly Gly Val Gly Lys 1 5 10 15 Ser Ala Leu Thr Ile
Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr 20 25 30 Asp Pro Thr
Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly 35 40 45 Glu
Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr 50 55
60 Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys
65 70 75 80 Val Phe Ala Ile Asn Asn Ser Lys Ser Phe Ala Asp Ile Asn
Leu Tyr 85 90 95 Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Asp Asp
Val Pro Met Val 100 105 110 Leu Val Gly Asn Lys Cys Asp Leu Pro Thr
Arg Thr Val Asp Thr Lys 115 120 125 Gln Ala His Glu Leu Ala Lys Ser
Tyr Gly Ile Pro Phe Ile Glu Thr 130 135 140 Ser Ala Lys Thr Arg Gln
Gly Val Glu Asp Ala Phe Tyr Thr Leu Val 145 150 155 160 Arg Glu Ile
Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Ser Asp Asp 165 170 175 Gly
Thr Gln Gly Cys Met Gly Leu Pro Cys Val Leu Met 180 185
51570DNACavia porcellus 51atgactgagt ataaactggt ggtggttgga
gcaggtggtg tcgggaaaag tgcactgacc 60atccagctaa ttcagaacca ctttgtcgat
gaatatgatc ccaccataga ggattcttac 120cgaaaacagg tggttataga
tggtgaaact tgtctgttgg atattctgga tacagctgga 180caagaggagt
acagtgccat gagagaccaa tacatgagga caggcgaagg cttcctctgt
240gtgtttgcca tcaataatag caaatcattt gcagatatta acctctacag
ggagcagatt 300aaacgagtaa aagactcaga tgatgtacct atggtgctgg
tagggaacaa gtgtgatttg 360ccaacaagga ctgttgacac aaaacaagcc
catgaactgg ccaagagtta cgggattcca 420ttcattgaaa cctcagccaa
gaccagacag ggtgttgaag atgcatttta cacactcgta 480agagaaatac
gccagtacag aatgaaaaaa ctcaacagca atgatgatgg gactcaaggt
540tgtatggggt tgccatgtgt ggtgatgtaa 57052189PRTCavia porcellus
52Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys 1
5 10 15 Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu
Tyr 20 25 30 Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val
Ile Asp Gly 35 40 45 Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala
Gly Gln Glu Glu Tyr 50 55 60 Ser Ala Met Arg Asp Gln Tyr Met Arg
Thr Gly Glu Gly Phe Leu Cys 65 70 75 80 Val Phe Ala Ile Asn Asn Ser
Lys Ser Phe Ala Asp Ile Asn Leu Tyr 85 90 95 Arg Glu Gln Ile Lys
Arg Val Lys Asp Ser Asp Asp Val Pro Met Val 100 105 110 Leu Val Gly
Asn Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys 115 120 125 Gln
Ala His Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr 130 135
140 Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val
145 150 155 160 Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser
Asn Asp Asp 165 170 175 Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val
Val Met 180 185 531220DNACavia porcellus 53gttccggggt cctcaacgtt
tctcagggtt gagattctat atccttttga agctggggcg 60gcagagcttg aggttcttgc
tggtgtgaaa tgactgagta taaactggtg gtggttggag 120caggtggtgt
cgggaaaagt gcactgacca tccagctaat tcagaaccac tttgtcgatg
180aatatgatcc caccatagag gattcttacc gaaaacaggt ggttatagat
ggtgaaactt 240gtctgttgga tattctggat acagctggac aagaggagta
cagtgccatg agagaccaat 300acatgaggac aggcgaaggc ttcctctgtg
tgtttgccat caataatagc aaatcatttg 360cagatattaa cctctacagg
gagcagatta aacgagtaaa agactcagat gatgtaccta 420tggtgctggt
agggaacaag tgtgatttgc caacaaggac tgttgacaca aaacaagccc
480atgaactggc caagagttac gggattccat tcattgaaac ctcagccaag
accagacagg 540gtgttgaaga tgcattttac acactcgtaa gagaaatacg
ccagtacaga atgaaaaaac 600tcaacagcaa tgatgatggg actcaaggtt
gtatggggtt gccatgtgtg gtgatgtaac 660aagatattta acaaagttct
atcagaaaag agccactttc aagctgcact gataccctgg 720tcctgacttc
cctggaggag aagtatccct gttgctctct tcatctcaga gaagctcctg
780ctgtttgtcc acctctcagt gtatgagcac agtctctgct tgagaacttc
tcagaataac 840tacctcctca cttggttgtc tgaccagaga aatgcacctc
ttgttaattc cccaataatt 900ttctgccctg ggctctcccc aacaaaaaac
aaacacttct gccatccaaa aagcaacttg 960gtctgaaaca gaaccaaact
gtagattgaa attctcttaa aaagtcttga gctctaaagt 1020tagcaaccgc
tggtgatttt tattttcctt tttatttttg aacttggaac tgacctatgt
1080tagattttgg agaaatgtca taaagtactg ttgtgccaag aagataatta
tgttgctgaa 1140tggttgattt atagtgttat cagctatatt ttacaaactg
gcatctgctc tgtattcata 1200aatacaaaaa tgaagccagg 122054189PRTCavia
porcellus 54Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val
Gly Lys 1 5 10 15 Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe
Val Asp Glu Tyr 20 25 30 Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys
Gln Val Val Ile Asp
Gly 35 40 45 Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln
Glu Glu Tyr 50 55 60 Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly
Glu Gly Phe Leu Cys 65 70 75 80 Val Phe Ala Ile Asn Asn Ser Lys Ser
Phe Ala Asp Ile Asn Leu Tyr 85 90 95 Arg Glu Gln Ile Lys Arg Val
Lys Asp Ser Asp Asp Val Pro Met Val 100 105 110 Leu Val Gly Asn Lys
Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys 115 120 125 Gln Ala His
Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr 130 135 140 Ser
Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val 145 150
155 160 Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Asn Asp
Asp 165 170 175 Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val Val Met
180 185 55 1307DNACanis familiaris 55tgattacgta gcgggcgggg
ccggaagtgc cgctccctag tgggggctgt tcatggcggt 60tccggggtct ccaacctttc
tcctagttgt ggtcctaaat acgtcggaag cggaggcggc 120gaagcttgag
gttcttgctg gtgtgaaatg actgagtaca aactggtggt ggttggagca
180ggtggtgttg ggaaaagcgc actgacaatc cagctaatcc agaaccactt
tgtagatgaa 240tatgatccca ccatagagga ttcttaccga aaacaggtgg
ttatagacgg tgaaacctgt 300ctgttggata tactggatac agctggtcaa
gaagagtaca gtgccatgag agaccaatac 360atgaggacag gcgaaggctt
cctctgtgta tttgccatca ataatagcaa atcatttgca 420gacattaacc
tctacaggga acagattaag cgagtaaaag attcagatga tgtacctatg
480gtgctagtag gaaacaagtg tgatttgcca acaaggacag ttgacacaaa
acaagcccat 540gaactggcca agagttatgg gattccattc attgaaacct
cagccaagac cagacagggt 600gtcgaggatg ccttttacac actggtaaga
gaaatacgtc agtaccgaat gaagaaactc 660aacagcagtg atgatgggac
tcaaggttgt atggggttac catgtgtggt gatgtaacaa 720gacactttta
aagttctagc atcagaaaag agccactgtc aagctgcact gacaccctgg
780tcctgacttc cctggaggag aagtattcct gttgctatct tcagtctcac
aaagaagctc 840ctgctacttc cccaactctc agtagatcag tacaatgttc
tctatttgag aagttctccg 900aacaactacc tcctcacttg gttgtctgac
cagagaaatg aacctcttgt tccttcccgc 960tgtttttcca ccctgaattc
tcccccaaca cacataaaca aacctctgcc atcccaggtt 1020tttcatctga
aaaataattc atgctctgaa acagagaaca aaactgtaga catgaaattc
1080tgtaggaaac aaggtcttga gctcaaaagt agcaactgct ggtgaccttt
ttttcccccc 1140tttttactgt tgaacttgga actatgttgg tttttggaga
aatgtcataa gttactgttt 1200tgctgagaat atagttaagt tgacatttgg
tttgtttgta atatcattag ctattttcta 1260taaattggca tctgctctgc
attcataaat acacgagtga attctga 130756189PRTCanis familiaris 56Met
Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys 1 5 10
15 Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr
20 25 30 Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile
Asp Gly 35 40 45 Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly
Gln Glu Glu Tyr 50 55 60 Ser Ala Met Arg Asp Gln Tyr Met Arg Thr
Gly Glu Gly Phe Leu Cys 65 70 75 80 Val Phe Ala Ile Asn Asn Ser Lys
Ser Phe Ala Asp Ile Asn Leu Tyr 85 90 95 Arg Glu Gln Ile Lys Arg
Val Lys Asp Ser Asp Asp Val Pro Met Val 100 105 110 Leu Val Gly Asn
Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys 115 120 125 Gln Ala
His Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr 130 135 140
Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val 145
150 155 160 Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Ser
Asp Asp 165 170 175 Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val Val
Met 180 185 573104DNAFelis catus 57aaaaaataaa taaatttaag aaaccatttt
aaaattatgc acagttgcag cctggaaaac 60ttaaggtggc gccttatagt atcaatctta
ggagctttat ttggtgcatt taacgcaact 120ggtaattgca aaatccactt
cgcctgtgta agtgaaaaat atagactgtt atcttgttgg 180ccctatgaaa
ttctgcactt ggtatttagc atatactcta ccttcattac tatctggcaa
240gatgttctgc cttagcactc agttgcattc ttttcctttt ctttcctgtt
cattatgctt 300taattctgag gaccatatga gggtagaata tattaaaaat
tacaaaaatt ataaaaattt 360gtataggcaa accatttcct taagttgatg
gccaaatgtt aaaatgttat ttttcatatc 420atttataatc ttgtcacagt
ccacttaacg aagtttggtt agatttcagt gaaaattatc 480ttccagagta
gttttttttt ttttttcctg ggattaggga ggggggtaac tttactgcaa
540ttagtatgta tggtgcagaa tttcatgcaa atgaggtgtg ccagcagtgt
ggtaatttaa 600tcgtatttaa acaaaaacaa acaaaaaaaa aacgaatgca
caaacttgct gctgcttaga 660tcactgcagc ttctaggacc cagtttcttt
tactgatttc aaaacaaaac aaaacaaaaa 720aataaaaaaa gttgtgcctg
aaatgaatct tgtttttttt ataagtagcc gcctggttcc 780tgtgtcctgt
gaaatacagg cacttgaccc ttggtgtagc ttctgttcga ctttatatca
840cgggaatgga ttggtctgat ttcttggccc tcatcttgaa ttggccacat
ccagggtccc 900tggccagtgg actgaaggct ttgtctaaga ggacaagggc
agctcagggg atgtggggga 960gggcgctttt atcttccccg ttgtcgtttg
aggttttgat cttctctggg taaagaggcc 1020gtttatcttt gtaaacacaa
aacatttttg ctttctccag ttttctgtta atggcgaaag 1080aatggaagcg
aataaagttt tactgatttt tgagactcta gcacctagcg ctttcatttt
1140tgaaacgtcc tgtgtgggag gggcgggtct gggtgcggcc cgccgcgtga
ctcctgagtc 1200gggggcccac gtggctgggg cggggactcg gacgccccgg
gcgccgactg attacgtagc 1260gggcggggcc ggaagtgccg ctccctagtg
ggggctgttc atggcggttc cggggtctcc 1320atcctttttc ccagttgttc
taaatcagtc ggaagcggag gcagcgaagt ttgaggttct 1380cgctggtgtg
aaatgactga gtacaaactg gtggtggttg gagcaggtgg tgttgggaaa
1440agcgcactga caatccagct aatccagaac cactttgtag atgaatatga
tcccaccata 1500gaggattctt accgaaaaca ggtggttata gacggtgaaa
cctgtctgtt ggacatactg 1560gatacagctg gtcaagaaga gtacagtgcc
atgagagacc aatacatgag gacaggcgaa 1620ggcttcctct gtgtatttgc
catcaacaat agcaaatcat ttgcagatat taacctttac 1680agggaacaga
ttaagcgagt aaaagactcc gatgatgtac ctatggtgct agtaggaaac
1740aagtgtgatt tgccaacaag gaccgtcgac acaaaacaag cccacgaact
ggccaagagt 1800tatgggattc cattcattga aacctcagcc aagaccagac
agggtgttga agatgccttt 1860tacacactgg taagagaaat acgtcagtac
cgaatgaaga aactcaacag cagtgatgac 1920gggactcaag gttgtatggg
gttaccgtgt gtggtgatgt aacaagatac ttttaaagtt 1980ctagcatcag
aaaagagcca ctgtcaagct gcactgacac cctggtcctg acttccctgg
2040aggagaagcg ttcctgttgc tattttcagt ttcacaaaga agctcctgct
atttccccaa 2100ctctccgtag atcagtacat tattctctgt ttgagaagtt
ctccgaataa ctacctcctc 2160acttggttgt ctgaccagag aaatgaacct
cttgttactc cccactgttt ttccaccctg 2220gttctccccc agcacatata
aacaaacctc ccaggttttt catctgaaaa gtaattcatg 2280ctctgaaaca
gagaaccaaa ctgtagacat gaaattctgt aggaaacaat gtcttgagct
2340ctaaagtagc aactgctggt gacttttttt tttttttttt cctttttact
gttgaacttg 2400gaactatgtt ggtttttgga gaaatgtcgt aagttactgt
tttgctgagt atatagttaa 2460gtttaccatt cggtttgttt gtaatgtcat
tggctatact ctgtacctgg catctgctct 2520gcattcataa atacaaaagt
gaattctgac ttttgagtct atcctagtgt tctcaacttc 2580cacataatta
aatctaactt ttgcagcaaa gtgccttttt cctagaagtg gtttgtagat
2640ttgctttata atactttggt ggaatagatg tctcaaaaac cattatacat
gaaaatgaat 2700gtctgagata cgtctatgat ctgtctacct ttgagggaaa
aatataccga cataatagca 2760gatgccatgt cttacgtgta tgaagttgga
tttccagaga cctgatttgg gtctcttcca 2820agagaaagat gaaactggaa
acaattatga ataacttcac ttaattttta cctaatctct 2880acttcggggt
gggagggcag ggagtaggtt accacttaca aaatatatgc aatttgtttc
2940ttctagctta ctgataatga acttccattc ttatttaaat ttaggtcata
tcctaaagct 3000ttacatttgc aggtgttcga aattgtaagt ttaatgcagt
tttatttaat agctatgatc 3060aatgattttc aagcctcaga tgtattaacg
gacacatttt cact 310458189PRTFelis catus 58Met Thr Glu Tyr Lys Leu
Val Val Val Gly Ala Gly Gly Val Gly Lys 1 5 10 15 Ser Ala Leu Thr
Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr 20 25 30 Asp Pro
Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly 35 40 45
Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr 50
55 60 Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu
Cys 65 70 75 80 Val Phe Ala Ile Asn Asn Ser Lys Ser Phe Ala Asp Ile
Asn Leu Tyr 85 90 95 Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Asp
Asp Val Pro Met Val 100 105 110 Leu Val Gly Asn Lys Cys Asp Leu Pro
Thr Arg Thr Val Asp Thr Lys 115 120 125 Gln Ala His Glu Leu Ala Lys
Ser Tyr Gly Ile Pro Phe Ile Glu Thr 130 135 140 Ser Ala Lys Thr Arg
Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val 145 150 155 160 Arg Glu
Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Ser Asp Asp 165 170 175
Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val Val Met 180 185
594283DNABos taurus 59ggccgctccc tagtgggggc tgttcatggc ggttccgggg
tctcccaaca attttcccgg 60ttgtggtcgt aatctatccg aagtggaggc agtggagcta
gaggttcttg ctggtgtgaa 120atgactgagt acaaactggt ggtggttgga
gcaggtggtg ttgggaaaag tgcactgaca 180atccagctaa tccagaacca
ctttgtagat gaatatgatc ccaccataga ggattcctac 240cgaaaacagg
tggttataga tggtgaaacc tgtctgttgg acatactgga tacagctgga
300caagaggagt acagtgccat gagagaccaa tacatgagga caggcgaagg
cttcctttgt 360gtgtttgcca tcaataatag caaatcattt gcagatatta
acctctacag ggaacagata 420aagcgtgtaa aggactcgga tgatgtacct
atggtgctag taggaaacaa gtgtgatttg 480ccaacaagga cagttgacac
aaaacaagcc catgaactgg ccaaaagtta tgggattcca 540ttcattgaaa
cctcagccaa gaccagacag ggtgttgaag atgcctttta cacactggta
600agagaaatac gtcagtaccg aatgaaaaag ctcaacagca gtgatgatgg
cactcaaggc 660tgtatggggt tgccgtgtgt ggtgatgtaa caagatactt
ttaaagttct cacatcagaa 720aagagccact gtcaagctgc actgacaccc
tggtcctgac ttccctggag gagaagtatt 780cctgttgcta tcttcagttt
caaaaagaag ctcctgctat ttccccaact ctcagtagat 840caatataata
ttctctattt gagaagttct caagaataac tacctcctca cttggttgtc
900tgaccagaga attgaacctc ttgttactcc cagtattttt ccaccctggg
ttctccccca 960gcacacacaa acgcacctct gccacccagg tttttcatct
gaaaagcaat taatactctg 1020aaacagagaa ccaaactgta gaaacatgaa
attctgtaga aaacaatgtc ttgagctcta 1080aagtagcaac tgctggtgat
tttttttttt tttttttcct ttttattgtt gaacttggaa 1140ctatgttggt
ttgtggagaa atgtcataaa ttactgtttt gctgagaata tagttaatgt
1200tgctctctgg tttgtttgta atgttatcag ctatattcta taaactggca
tctactctgt 1260atttagaaat acaaaaatga atactgacct tttgagtcta
ccctcatctt ctcgactttc 1320ttgtaattaa atgtaacttt cacgatgaag
tgccttttgc ctgggagtga ctcgtagact 1380tcctttaaaa tacttcagtg
gaatagatgt ctcagaaact gttatacata agaataaatg 1440tctgagatat
gtctatgacc catctagctt tgagggaaag atataccaat atgatagcag
1500atgccatttc ttacatctat aacgttgatt ttctggagac ctattttggg
gctctccgag 1560agaaagatga gactataaat gattaggaat aatttcactt
aatttttaca taacctccac 1620tttttgtttt gtagtttact acctgcaaaa
catataattt gattcctttt agcttacaga 1680taatctaatg ttaaatgaac
ttccattcat attttaattt ggatcatatc aggaagtcta 1740catttgcagg
tgttcaaaaa ttgtaaaagt gtgatgcagt tttatttaat agctgtgatc
1800aatgattttc aagcctcaaa tatgttaata gacacatttt cactgtatat
catggtatta 1860ataattattg atgtatataa ttgtccttgg tccccttctc
tgttcatcac ctcatggcaa 1920tggcttgatt aattatttca gctgagtaaa
gcatggtgct aatagaccag ggtcacagtg 1980tcaaaacttc agtgagccag
taagcatcac agagaaagaa attctttcac atttgctcac 2040cattaactcc
agctaatagt tttgccagat gtgtgtggtt agtcctgcaa ggaaaggaga
2100agtcagttaa tacaaattct taaccaggac tggaaaaact tgttttcctg
agaagggtca 2160gcttagaagt ctttatctgg actctatttt tagccacatg
gaaatcaaat taagctgatc 2220ttttttctca agtttttgag agtgaggatg
cctcagatca acatttttaa aatattcttt 2280attcttacgt tcttttaagg
gtttaaaaca acgttgagta attagtctgg gcataccagg 2340taacaagctg
ataagtttgt gctgaacaag aagtagcctt tggattgaaa ttgctgtttt
2400gagaagggat agaaaatata attaataatt atgagacttg acttttctat
ttgcagataa 2460tatcctgata attctgatga aaatagactt ggataatttt
tgataaaaga atcgttccaa 2520aatggccact tgctgttctt gtcttctaat
gtgtaaatac ttactgaggt cctcttctaa 2580tatgagttgt catttattaa
gcaaattcca cattgccttg aaatgaattc ggaagagaag 2640aaaaagtcat
agtataccca gagaatgaaa aatccagaga attgtgctcc ttagtgttaa
2700ttctgaagcc ttcgtagtcc acacccatag acagaaactc tctgccactt
tgcttctgct 2760cctcttggag cattgcgctg tcatttcctt gaggatagat
tgaggcttgt caactcagtt 2820gtattgtctt cctcctcttc ctcttgtctg
tgtgactgac agtgtgactc ttactaatgt 2880cagatgcggg gatgcgggga
ggtggggggg agtagctcat tttaggctct tgcacccttt 2940accgttgtat
gtgtgtgtct tttagttttc tcaagaatgt tctaagcaca gaagtatcta
3000aatggggcca aaattcagac ttgaaaatgt tcttttaata gcttcttaaa
aagttacact 3060ttggtgtgaa ttttggcagg atagagtgac aaactcttaa
acgctgaata acttcagtta 3120gtgtgttata gtttttagaa tatgtttgtg
attgctgaaa acaattatag tttacctcaa 3180aatctgaaag tctctttccc
caagttaagt gcctggccag ctgtcaaaga ttacatatta 3240ctttatgttt
gtttgttttt taaaggttgc acattcaaga ttgtgaaaat aaggtgttct
3300gtctgaaagc taccatgcct gtctgtaaat gaatccactg agtgctgtac
ttgttccaac 3360agcttactac agaatgctac ttggtaatat catactcgtt
acagttttca cttcaggagt 3420gtactaggta gaatgatcct gtgtgtattg
tagtgggctc catgtttagt cttttcagca 3480tcctttaaac tgctgtgaat
ttttgtcttg acttgaaagc aaggatagag aaacacttta 3540aagagatact
ttgggttttt ttccattcca gaattggtga gcatagttag attttgcttt
3600acatttacag tcatgaactc ttaagctggc agctacaacc aagaaccaaa
agagggtgca 3660ttctgcttct tgtaattcat ctttgctaat aaattatgag
aagcaaagat aattaattag 3720agaaactatt ttatttgggt ggtttctata
aacaagggac tataattctt aaacattatt 3780tttcattttt gctgtttctt
taagaaacct aatgtgccac aacattattt taaggtgttt 3840cttaaaagaa
ttgtttttaa aagtgttctc attttcagag taattgtaga tatatttcaa
3900aatataactg ataattttta aaggcctgag tactgaccta agaagcagtt
gtatgaattc 3960tctgggggga agggaggagc tcagtgaaag ttgtatgact
tttatatttc tgtgccatca 4020aataaaggta aaaatgtctt ttgtgcagtt
ttgctgttca aacagaaact attggcctcc 4080ttggccctaa atgaaagggc
tggtatttta agttgactat tttattgtaa attaatccat 4140cttaattttt
ttaaatttgg ttgaatgttc tcttgttaaa tgtttaaaaa ataaaaactg
4200gaagttcttt gcttagtcat aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa 4260aaaaaaataa aaaaaaaaaa aaa 428360189PRTBos taurus
60Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys 1
5 10 15 Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu
Tyr 20 25 30 Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val
Ile Asp Gly 35 40 45 Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala
Gly Gln Glu Glu Tyr 50 55 60 Ser Ala Met Arg Asp Gln Tyr Met Arg
Thr Gly Glu Gly Phe Leu Cys 65 70 75 80 Val Phe Ala Ile Asn Asn Ser
Lys Ser Phe Ala Asp Ile Asn Leu Tyr 85 90 95 Arg Glu Gln Ile Lys
Arg Val Lys Asp Ser Asp Asp Val Pro Met Val 100 105 110 Leu Val Gly
Asn Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys 115 120 125 Gln
Ala His Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr 130 135
140 Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val
145 150 155 160 Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser
Ser Asp Asp 165 170 175 Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val
Val Met 180 185 614825DNAGallus gallus 61gcgccgggac cggaagccgg
aagctttgca gaagggtgtt ccgcgttcgc ggtgcgggag 60cggtcagccg gggtggcggg
gctggggccg gccggggcag gcggctccgc gctccgcact 120gggccgctgg
gagggcgatg actgaataca agctggtggt ggtgggagct ggcggcgtcg
180ggaagagcgc gttgaccatc cagctcatcc agaaccactt cgtggacgag
tacgacccca 240ccatcgagga ttcgtacaga aagcaggttg tcatcgatgg
agagacgtgc ttgttggaca 300ttctggacac tgcaggacag gaagaataca
gtgctatgcg tgatcagtac atgagaactg 360gggaaggatt cctttgtgtg
tttgccatta acaacagtaa atcattcgct gatattaacc 420tttacagaga
gcaaatcaag agagtgaaag attcagacga tgtgccaatg gtgctggtgg
480ggaataagtg cgatttgcca acaaggacag tagacaccaa acaggctcaa
gagttagcaa 540aaagctacgg cattcccttc atagagacat cagccaaaac
gagacagggt gtggaagatg 600cgttttacac actggtgagg gagattcggc
agtaccggat gaaaaagctc aacagcaacg 660aagatgggaa tcagggctgt
atggggttgt cctgcattgt gatgtgataa gatgccaggt 720tcagatgtag
ctgctggaca agtctcgatg ctactgtatt gtgtctcatg ctgatgccct
780gcagtatttt ggtgccagcg accagactct tggtaccagt taattagctc
aggatccttt 840cctgtgctcc atctgaagaa aacatctctg gtatctacct
ccttgctcag ctcacagagc 900agtcatatct cttggtgtac tgggattctt
ttctagctgt gttgtctggg tttgttcaag 960aagaaaacca gtcacaagaa
aagtgaatta cagagactaa atgctgtgaa aaagatcaca 1020ctttacctcc
agagtaaaag ctagaagtgg cgtttgaccc ctttgcattg gattcagatt
1080tgcggtgttg tcagaggagt ggcagaagta attttgccat tacaaaggtt
tctgtcacca 1140gtcggattgg tatctgctgt ctgtgcaccc acacagtgta
tctgcaacat ctgcattgtg 1200ccagaagtat cacttaactg atgaactgat
cctttatttt tctgtaataa aaaggagata 1260tctttgctaa cttaagtgcc
tgtttgctca gaaggttgga ggttgtatgc tgttcccttg 1320ggctgaggag
aaccccaagg atgaatttct tgggtgctca ttgtcttgag caggcaagtt
1380ttgtgtgggt gatctctttt catggcagga tattaaaatg ggaatttgta
gtctggaaga
1440tggagcagct gtttgtgaga ctcttgagtt agggagagaa atgtatacca
cgtctgttct 1500cgatccatca gaatggatcc atccacctct ttgtgtgtgg
aactgtgtat agtctgtatt 1560ggttttctac agcacttgga tctctttgga
ccaaattagc gagctgttca ttttaacata 1620actgccagta tttatagaca
atttcttacg gacagataat gaatttagaa actggaggtt 1680actttgggca
gctgttcctc agctctgtct gtaacttgca aattattctg agttattttc
1740tgcagaacct ccttccttat cacgggagga gcctgggagt tgaggttgac
tgtaattggg 1800tcaatggttg tcacagactt aaggtgtcca ggctgattgg
aggaggcact gagccctaac 1860agagcactga gctgacttct aattgcagca
tccttgcaaa atgaggaagg gagttcagtg 1920atgtctgcac tgaagatgta
tgatacactg atagcagttc tgggtatgtt gtaacagctt 1980caaagtagaa
ccgcagtact gcgtgagctg tgtgacttct tcctagaaca cagcactgtc
2040accccatatg gttgggacgt gcaggtgaga ccaacaccta ccaggttccc
tggcgtaccg 2100tggccttctc agttcttgtg ccagtgatac tgggttctgt
tctgtggtgt cagacagcgt 2160cctgtagcaa agctgaattc ccacttagtc
tggtgagaga ataaagagcc atcagccaac 2220agagggagcg ttcattctgc
tggagcagtg cgagctgtaa gcattacgag aggcgtagtt 2280tcagtttgtt
gcagtcaggt tcctatattt tcaaagctga aatcagaaat aagtaaatac
2340ggagaaaata agctgttgct tttaatgctc tttcctccac taattgtact
cttaattttc 2400ttcttgggag gccgaggatc catctgcata actttagctg
tgatgctcca gataagtgtt 2460tagaattcat tttatctttg actgatggga
ctgataagaa gttaacgcac aatattttta 2520catacaacat cgttttccag
tgacctcctg agcggtggga agcattatgg gatagcaccg 2580gctgtgactc
gagttcattt gaaggcgatc tcttgcctgc aggttaaatg ggacggagtc
2640agaatcactg tgagccgtct gtaatcagca aacagtctgt gggcttttct
tactgtgttc 2700tctctgtttg ccttagtttg gtgcaggaag agttccttgt
gacagcgtcc tttgaggtgt 2760gttgcaggag ctgaccattt gctccttgag
ctgtgtgatg aactgttgtc cacttaatgg 2820agttacagaa gcagcttctg
ggagtcgcat ctggtcgcat acattcagtg ttttgggaag 2880ctgtcagtgt
ggtgtttgca ctgtgtttga atggtgttca tggtgggtct gttatgctcc
2940tggatgattt ggggagatgt ggggctgctt ccgtggcaga caggatcagc
tcagggcgct 3000gctgcctatg gctgtgggaa acctcacagt tggtgtttga
atagtggcca agtatgtcaa 3060ttaaaaatac attttgaagg gaggtttgtc
atagctctgt actttggcat gctctgctta 3120ctgaaaacat actagctgta
gctcaaaaaa agttgtgaat cctcagaata atacaggagc 3180tggcaattgt
ggctgctttc tctttgtgtt ccttttctct tgggttggat gaagctttaa
3240aaaggaagga gccctggtga gggttggtca gtgtgcattt cattcttgga
accagagagg 3300aagttgcatc aactttcagg acgctgcaga gctcacttgc
acaggtggtg ctccagtcta 3360tgtgattttt ggggtcaaat cttgagatga
tcttacaaaa tcagattttg tacccatcat 3420gagcatgagg tgagtggttg
tgctcggttt ctagctgcat gtatgtatac agacacgtgt 3480atgcagacat
gtctatgtgt gagtagttcg agtcagtcaa ggttactggc agcacctaaa
3540gcgtatgcac cacataatgc atgcaggcaa aagtcctatc ttaggagcca
tctcttcatg 3600ggtttgggtt tatataggca gtatttttaa acagaatatc
cgaagcactt tctggagttc 3660tgtggtaatg cagtgacacc tatttggatg
aaggaagatg tgtctgagga gcacgtaagc 3720agatttgctg ccctaacaga
gaggttttgg taaccgtgga aaaggttttc tcctggatct 3780gtgtgtgctc
ttggtgagct gcaatccatg acagggcaca accagatgag aaggaaaccc
3840ggccatccca tgcttgagca cagctctgac tcagtagttc caccagatgt
gccctttcag 3900tcaaagtgtt ctgatctctt agagctttct gtagttcaag
ttaccactca ctctccagct 3960tgctcggtta atgtctgttg gcggcgttga
gttggacttg ggaaaggtgt gtgtggtagg 4020aacaagcaga gtgtgatgtg
cttctgttat caggacttaa gctagagtgg ttggcagata 4080ggaaatgcag
ctattccttg aaagcaagca gatcatggat ggtcagccaa actgccctgg
4140ctttggtggg agctgcactg cagaaggacc aaaccccaac aagatttggc
acatttgttt 4200agaagataag cacagatggt tttgcacaag gcagctcctc
ataatggtgg ctttgtagat 4260ttagtccaaa tgttcttatt tagatctagc
agcacatcac tgtgtccgtg cccatctaac 4320ctcgctatcc taagtagagc
agaccccaaa caaccttgtt caaaaactac cagtgcaaat 4380aactgaacta
aatatttgtt actgctgact gagaacagct gttcgagtgt agcattgtgg
4440cttgttaatg tgagtgcccc aactctatgg tcttattaaa gaaacccaaa
cattgctcag 4500attttgttct tattgtcatc ataagacttg aatagtgatg
gtaatgctta cgtagacgtg 4560tcttgtgagt gcacttcagt gatttagaaa
gaactggatt tcaagcaact ttggacctgt 4620ggggggaggg agattaatga
aggtttgaat cacattctaa ttctatgtac agtccttcat 4680tactccacaa
gcctaaatcc tatacagcct ccaggatagc tggaaactgt tgagatctgg
4740actttttttt tttaatccaa gggctaactt gttgtaactt ggtataatta
tctgctttcg 4800gaaatgcatc tctgttggtt tgaaa 482562189PRTGallus
gallus 62Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val
Gly Lys 1 5 10 15 Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe
Val Asp Glu Tyr 20 25 30 Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys
Gln Val Val Ile Asp Gly 35 40 45 Glu Thr Cys Leu Leu Asp Ile Leu
Asp Thr Ala Gly Gln Glu Glu Tyr 50 55 60 Ser Ala Met Arg Asp Gln
Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys 65 70 75 80 Val Phe Ala Ile
Asn Asn Ser Lys Ser Phe Ala Asp Ile Asn Leu Tyr 85 90 95 Arg Glu
Gln Ile Lys Arg Val Lys Asp Ser Asp Asp Val Pro Met Val 100 105 110
Leu Val Gly Asn Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys 115
120 125 Gln Ala Gln Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu
Thr 130 135 140 Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr
Thr Leu Val 145 150 155 160 Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys
Leu Asn Ser Asn Glu Asp 165 170 175 Gly Asn Gln Gly Cys Met Gly Leu
Ser Cys Ile Val Met 180 185 632603DNAHomo sapiens 63aggcgaggct
tccccttccc cgcccctccc ccggcctcca gtccctccca gggccgcttc 60gcagagcggc
taggagcacg gcggcggcgg cactttcccc ggcaggagct ggagctgggc
120tctggtgcgc gcgcggctgt gccgcccgag ccggagggac tggttggttg
agagagagag 180aggaagggaa tcccgggctg ccgaaccgca cgttcagccc
gctccgctcc tgcagggcag 240cctttcggct ctctgcgcgc gaagccgagt
cccgggcggg tggggcgggg gtccactgag 300accgctaccg gcccctcggc
gctgacggga ccgcgcgggg cgcacccgct gaaggcagcc 360ccggggcccg
cggcccggac ttggtcctgc gcagcgggcg cggggcagcg cagcgggagg
420aagcgagagg tgctgccctc cccccggagt tggaagcgcg ttacccgggt
ccaaaatgcc 480caagaagaag ccgacgccca tccagctgaa cccggccccc
gacggctctg cagttaacgg 540gaccagctct gcggagacca acttggaggc
cttgcagaag aagctggagg agctagagct 600tgatgagcag cagcgaaagc
gccttgaggc ctttcttacc cagaagcaga aggtgggaga 660actgaaggat
gacgactttg agaagatcag tgagctgggg gctggcaatg gcggtgtggt
720gttcaaggtc tcccacaagc cttctggcct ggtcatggcc agaaagctaa
ttcatctgga 780gatcaaaccc gcaatccgga accagatcat aagggagctg
caggttctgc atgagtgcaa 840ctctccgtac atcgtgggct tctatggtgc
gttctacagc gatggcgaga tcagtatctg 900catggagcac atggatggag
gttctctgga tcaagtcctg aagaaagctg gaagaattcc 960tgaacaaatt
ttaggaaaag ttagcattgc tgtaataaaa ggcctgacat atctgaggga
1020gaagcacaag atcatgcaca gagatgtcaa gccctccaac atcctagtca
actcccgtgg 1080ggagatcaag ctctgtgact ttggggtcag cgggcagctc
atcgactcca tggccaactc 1140cttcgtgggc acaaggtcct acatgtcgcc
agaaagactc caggggactc attactctgt 1200gcagtcagac atctggagca
tgggactgtc tctggtagag atggcggttg ggaggtatcc 1260catccctcct
ccagatgcca aggagctgga gctgatgttt gggtgccagg tggaaggaga
1320tgcggctgag accccaccca ggccaaggac ccccgggagg ccccttagct
catacggaat 1380ggacagccga cctcccatgg caatttttga gttgttggat
tacatagtca acgagcctcc 1440tccaaaactg cccagtggag tgttcagtct
ggaatttcaa gattttgtga ataaatgctt 1500aataaaaaac cccgcagaga
gagcagattt gaagcaactc atggttcatg cttttatcaa 1560gagatctgat
gctgaggaag tggattttgc aggttggctc tgctccacca tcggccttaa
1620ccagcccagc acaccaaccc atgctgctgg cgtctaagtg tttgggaagc
aacaaagagc 1680gagtcccctg cccggtggtt tgccatgtcg cttttgggcc
tccttcccat gcctgtctct 1740gttcagatgt gcatttcacc tgtgacaaag
gatgaagaac acagcatgtg ccaagattct 1800actcttgtca tttttaatat
tactgtcttt attcttatta ctattattgt tcccctaagt 1860ggattggctt
tgtgcttggg gctatttgtg tgtatgctga tgatcaaaac ctgtgccagg
1920ctgaattaca gtgaaatttt ggtgaatgtg ggtagtcatt cttacaattg
cactgctgtt 1980cctgctccat gactggctgt ctgcctgtat tttcgggatt
ctttgacatt tggtggtact 2040ttattcttgc tgggcatact ttctctctag
gagggagcct tgtgagatcc ttcacaggca 2100gtgcatgtga agcatgcttt
gctgctatga aaatgagcat cagagagtgt acatcatgtt 2160attttattat
tattatttgc ttttcatgta gaactcagca gttgacatcc aaatctagcc
2220agagcccttc actgccatga tagctggggc ttcaccagtc tgtctactgt
ggtgatctgt 2280agacttctgg ttgtatttct atatttattt tcagtatact
gtgtgggata cttagtggta 2340tgtctcttta agttttgatt aatgtttctt
aaatggaatt attttgaatg tcacaaattg 2400atcaagatat taaaatgtcg
gatttatctt tccccatatc caagtaccaa tgctgttgta 2460aacaacgtgt
atagtgccta aaattgtatg aaaatccttt taaccatttt aacctagatg
2520tttaacaaat ctaatctctt attctaataa atatactatg aaataaaaaa
aaaaggatga 2580aagctaaaaa aaaaaaaaaa aaa 260364393PRTHomo sapiens
64Met Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro Ala Pro Asp 1
5 10 15 Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu
Ala 20 25 30 Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln
Gln Arg Lys 35 40 45 Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys
Val Gly Glu Leu Lys 50 55 60 Asp Asp Asp Phe Glu Lys Ile Ser Glu
Leu Gly Ala Gly Asn Gly Gly 65 70 75 80 Val Val Phe Lys Val Ser His
Lys Pro Ser Gly Leu Val Met Ala Arg 85 90 95 Lys Leu Ile His Leu
Glu Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile 100 105 110 Arg Glu Leu
Gln Val Leu His Glu Cys Asn Ser Pro Tyr Ile Val Gly 115 120 125 Phe
Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile Cys Met Glu 130 135
140 His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys Lys Ala Gly Arg
145 150 155 160 Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile Ala Val
Ile Lys Gly 165 170 175 Leu Thr Tyr Leu Arg Glu Lys His Lys Ile Met
His Arg Asp Val Lys 180 185 190 Pro Ser Asn Ile Leu Val Asn Ser Arg
Gly Glu Ile Lys Leu Cys Asp 195 200 205 Phe Gly Val Ser Gly Gln Leu
Ile Asp Ser Met Ala Asn Ser Phe Val 210 215 220 Gly Thr Arg Ser Tyr
Met Ser Pro Glu Arg Leu Gln Gly Thr His Tyr 225 230 235 240 Ser Val
Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu Val Glu Met 245 250 255
Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu 260
265 270 Leu Met Phe Gly Cys Gln Val Glu Gly Asp Ala Ala Glu Thr Pro
Pro 275 280 285 Arg Pro Arg Thr Pro Gly Arg Pro Leu Ser Ser Tyr Gly
Met Asp Ser 290 295 300 Arg Pro Pro Met Ala Ile Phe Glu Leu Leu Asp
Tyr Ile Val Asn Glu 305 310 315 320 Pro Pro Pro Lys Leu Pro Ser Gly
Val Phe Ser Leu Glu Phe Gln Asp 325 330 335 Phe Val Asn Lys Cys Leu
Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu 340 345 350 Lys Gln Leu Met
Val His Ala Phe Ile Lys Arg Ser Asp Ala Glu Glu 355 360 365 Val Asp
Phe Ala Gly Trp Leu Cys Ser Thr Ile Gly Leu Asn Gln Pro 370 375 380
Ser Thr Pro Thr His Ala Ala Gly Val 385 390 652151DNARattus
norvegicus 65cggccgcgcg ctccctgctg agttgcaggc tgtttcccgg ctgcaagatg
cccaagaaga 60agccgacgcc catccagctg aacccggccc ccgatggctc cgcggttaac
gggaccagct 120cggccgagac caacctggag gccttgcaga agaagctgga
ggagctggag ctggacgagc 180agcagcggaa gcgccttgag gcctttctga
cgcagaagca gaaggtggga gagttgaagg 240atgatgactt tgagaagatc
agtgaactgg gggctggcaa tggtggagtg gtgttcaagg 300tctcccacaa
gccatctggc ctggttatgg ctaggaagct aattcacctg gagatcaaac
360ccgcaatccg gaaccagatc atccgggagc tgcaggtgct gcatgagtgc
aactccccgt 420acatagtggg cttctacggg gccttctaca gtgacggcga
gatcagcatc tgcatggagc 480acatggatgg tgggtccttg gatcaagtgc
tgaagaaagc tggaagaatt cctgagcaaa 540ttttaggaaa agtcagcatc
gctgtgataa aaggcctgac atatctacga gagaagcaca 600agattatgca
cagagatgtc aagccttcca acattctagt gaactcacgt ggggagatca
660aactctgcga ttttggggtc agcgggcagc taattgactc catggccaac
tccttcgtgg 720gaacaaggtc ctacatgtcg cctgagagac tccaggggac
tcactactct gtgcagtcgg 780acatctggag catggggctc tctctggtgg
agatggcagt tggaagatac cccattcctc 840ctcctgatgc caaggagctg
gagctgctgt ttggatgcca ggtggaagga gacgcggccg 900aaacgccacc
caggccaagg acccctggga ggcccctcag ctcatatgga atggatagcc
960gacctcccat ggcaattttt gagttgttgg attacatcgt caatgagcct
cctccaaaac 1020tgcccagtgg agtattcagt ctggaatttc aggattttgt
gaataagtgc ttaataaaga 1080accctgcaga gagagcagat ctgaagcagc
tcatggtaca tgctttcatc aagagatctg 1140atgccgagga ggtagacttc
gcaggctggc tctgctccac cattgggctt aaccagccca 1200gcacaccaac
ccacgctgcc agcatctgag cctttgggaa gcagcagaga ggaatcctct
1260gcccagtggc atgccatgtt gctttcaggc ctctcccatg cttgtctatg
ttcagatgtg 1320catctcatct gtgacaaagg atgaagaaca cagcatgtgc
caaatcgtac ttgtgtcatt 1380tttaatattg tctttatcgc tatggttact
cccctaagtg gattggcttt gtgcttgggg 1440ctatttgtct gttcatcaaa
tacatgccag gttgaactac agtgaaaccc tggtgacctg 1500ggtggtcttc
ttactgatgt ttgcgctgct gttcatcgtg actcactagc tggctgcctg
1560tattgtcagg attctcggac ccttggtact tcactcttgc tggtgacctc
tcagtctgag 1620gagagggggc cttctgagac ccttcacagg cagtgcatgc
atgaaaagca tgctttgctg 1680ctactgaaat gagcaccaga acgtgtacat
catggtattt tatttttgct tttggtatag 1740aactcagcag ttcccattta
aaaaaaaaat ctaaccagag cccatcactg ccatgatagc 1800tggggcttca
gtctgtctac tgtggtgatt tttagacttc tggttgtatt tctatattta
1860tttttaaata tactgtgtgg gatacttagt ggtatatgtc tctgagtttg
gattagtgtt 1920tctaaattgg tagttatttt gaatgtcaca aatggattaa
ggaatcaacg tatcaagagt 1980tctatctttc ttccagtcta agtaccaatg
ctattgtaaa cgtgtatagt gcctacaaat 2040tgtatgaaaa cccttttaac
cactttactc aagatgttta tcaaatctaa tctcttattc 2100taataaaaat
actatcaagt taaagtaaaa aaaaaaaaaa aaaaaaaaaa a 215166393PRTRattus
norvegicus 66Met Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro
Ala Pro Asp 1 5 10 15 Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu
Thr Asn Leu Glu Ala 20 25 30 Leu Gln Lys Lys Leu Glu Glu Leu Glu
Leu Asp Glu Gln Gln Arg Lys 35 40 45 Arg Leu Glu Ala Phe Leu Thr
Gln Lys Gln Lys Val Gly Glu Leu Lys 50 55 60 Asp Asp Asp Phe Glu
Lys Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly 65 70 75 80 Val Val Phe
Lys Val Ser His Lys Pro Ser Gly Leu Val Met Ala Arg 85 90 95 Lys
Leu Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile 100 105
110 Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr Ile Val Gly
115 120 125 Phe Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile Cys
Met Glu 130 135 140 His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys
Lys Ala Gly Arg 145 150 155 160 Ile Pro Glu Gln Ile Leu Gly Lys Val
Ser Ile Ala Val Ile Lys Gly 165 170 175 Leu Thr Tyr Leu Arg Glu Lys
His Lys Ile Met His Arg Asp Val Lys 180 185 190 Pro Ser Asn Ile Leu
Val Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp 195 200 205 Phe Gly Val
Ser Gly Gln Leu Ile Asp Ser Met Ala Asn Ser Phe Val 210 215 220 Gly
Thr Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly Thr His Tyr 225 230
235 240 Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu Val Glu
Met 245 250 255 Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys
Glu Leu Glu 260 265 270 Leu Leu Phe Gly Cys Gln Val Glu Gly Asp Ala
Ala Glu Thr Pro Pro 275 280 285 Arg Pro Arg Thr Pro Gly Arg Pro Leu
Ser Ser Tyr Gly Met Asp Ser 290 295 300 Arg Pro Pro Met Ala Ile Phe
Glu Leu Leu Asp Tyr Ile Val Asn Glu 305 310 315 320 Pro Pro Pro Lys
Leu Pro Ser Gly Val Phe Ser Leu Glu Phe Gln Asp 325 330 335 Phe Val
Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu 340 345 350
Lys Gln Leu Met Val His Ala Phe Ile Lys Arg Ser Asp Ala Glu Glu 355
360 365 Val Asp Phe Ala Gly Trp Leu Cys Ser Thr Ile Gly Leu Asn Gln
Pro 370 375 380 Ser Thr Pro Thr His Ala Ala Ser Ile 385 390
672387DNAMus musculus 67agtccctcac tgggacgtct gtgcgcggcg tctcggagcg
ccggagcagc ggtggccgca 60ctttctccaa gctggggctg tagctgagct gtgggtagtg
cgcagggagc cgtccgagcc 120cgaggaaccg gtgtgctgag gcgagagttc
ccggccggcg agcgcgcgca gctggttctc 180cgcgtgggtt gggcggaggg
tcccaggagc gcggcgttga tcgagccgcc ccgactctgg 240gcagagccga
gggaggaagc gagaagcggc cgcgcgctcc ctgctgagtt gcaggctctt
300tcccggctgc aagatgccca
agaagaagcc gacgcccatc cagctgaacc cggcccccga 360tggctcggcg
gttaacggga ccagctcggc cgagaccaac ctggaggcct tgcagaagaa
420gctggaggag ctggagcttg acgagcagca gcggaagcgg ctcgaggcct
ttctgacgca 480gaagcagaag gtgggggaac tgaaggatga tgactttgag
aagatcagcg aactgggagc 540tggcaacggt ggagtggtct tcaaggtctc
ccacaagcca tctggcctgg ttatggctag 600aaagctgatc cacctggaga
tcaaacccgc aatccggaac cagatcatcc gggagctgca 660ggtactgcac
gagtgcaact ccccgtacat cgtgggcttc tacggggcct tctacagcga
720cggcgagatc agcatctgca tggagcacat ggatggtggg tccttggatc
aagttctgaa 780gaaagctgga agaattcctg agcaaatttt aggaaaagtt
agcattgctg tgataaaagg 840cctgacctat cttcgggaga agcacaagat
tatgcacaga gatgtcaagc catccaacat 900tctagtgaac tcacgtgggg
agatcaaact ctgtgatttt ggggtcagcg ggcagctaat 960tgactctatg
gccaactcct tcgtgggcac gagatcctac atgtcgcctg agagactcca
1020ggggactcac tactctgtgc agtcggacat ctggagcatg gggctctctc
tggtggagat 1080ggcagttggg agatacccca ttcctcctcc tgatgccaag
gagctggagc tactgtttgg 1140atgccatgtg gaaggagacg cagccgaaac
accacccagg ccaaggaccc ctgggaggcc 1200tctcagctca tatggaatgg
acagccgacc tcccatggca atttttgagt tgttggatta 1260cattgtcaat
gagcctcctc caaaactgcc cagtggagta ttcagtctgg agtttcagga
1320ttttgtgaat aaatgcttaa taaagaaccc tgcagagaga gcagatctga
agcagctcat 1380ggtacatgct ttcatcaaaa gatctgacgc cgaggaggta
gacttcgcag gctggctctg 1440ctccaccatt gggcttaacc agcccagcac
accaacccac gctgccagca tctgagcctt 1500taggaagcag caaagaggaa
ttctctgccc agtggcatgc catgttgctt tcaggcctct 1560cccatgcttg
tctatgttca gacgtgcatc tcatctgtga caaaggatga agaacacagc
1620atgtgccaaa ttgtacttgt gtcattttta atatcattgt ctttatcact
atggttactc 1680ccctaagtgg attggctttg tgcttggggc tatttgtctg
ttcatcaaac acatgccagg 1740ctgaactaca gtgaaaccct agtgacctgg
gtggtcgttc ttactgatgt ttgcactgct 1800gttcatcgtg actcactagc
tggctgcctg tattgtcagg attctcggac cttggtactt 1860cactcttgct
ggtgacctct cagtctgaga gggagccttg tgagaccctt cacaggcagt
1920gcatgcatgg aaagcatgct ttgctgctac tgaaatgagc atcagaacgt
gtacgtcatg 1980gtatttttat tttttgcttt tggtatagaa ctcagcaatt
cccatcaaaa aaacctaagc 2040agagcccatc actgccatga tagctgggct
tcagtctgtc tactgtggtg atttttagac 2100ttctggttgt atttctatat
ttatttttaa atatacagtg tgggatactt agtggtgtgt 2160gtctctaagt
ttggattagt gtttctaaat tggtggttat tttgaatgtc acaaatggat
2220taaagcatca atgtatcaag agttctatct ttcttccagt ctaagtacca
atgctattgt 2280aaacaacgtg tatagtgcct acaaattgta tgaaacccct
tttaaccact ttaatcaaga 2340tgtttatcaa atctaatctc ttattctaat
aaaaatacta tcaagtt 238768393PRTMus musculus 68Met Pro Lys Lys Lys
Pro Thr Pro Ile Gln Leu Asn Pro Ala Pro Asp 1 5 10 15 Gly Ser Ala
Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu Ala 20 25 30 Leu
Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys 35 40
45 Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys Val Gly Glu Leu Lys
50 55 60 Asp Asp Asp Phe Glu Lys Ile Ser Glu Leu Gly Ala Gly Asn
Gly Gly 65 70 75 80 Val Val Phe Lys Val Ser His Lys Pro Ser Gly Leu
Val Met Ala Arg 85 90 95 Lys Leu Ile His Leu Glu Ile Lys Pro Ala
Ile Arg Asn Gln Ile Ile 100 105 110 Arg Glu Leu Gln Val Leu His Glu
Cys Asn Ser Pro Tyr Ile Val Gly 115 120 125 Phe Tyr Gly Ala Phe Tyr
Ser Asp Gly Glu Ile Ser Ile Cys Met Glu 130 135 140 His Met Asp Gly
Gly Ser Leu Asp Gln Val Leu Lys Lys Ala Gly Arg 145 150 155 160 Ile
Pro Glu Gln Ile Leu Gly Lys Val Ser Ile Ala Val Ile Lys Gly 165 170
175 Leu Thr Tyr Leu Arg Glu Lys His Lys Ile Met His Arg Asp Val Lys
180 185 190 Pro Ser Asn Ile Leu Val Asn Ser Arg Gly Glu Ile Lys Leu
Cys Asp 195 200 205 Phe Gly Val Ser Gly Gln Leu Ile Asp Ser Met Ala
Asn Ser Phe Val 210 215 220 Gly Thr Arg Ser Tyr Met Ser Pro Glu Arg
Leu Gln Gly Thr His Tyr 225 230 235 240 Ser Val Gln Ser Asp Ile Trp
Ser Met Gly Leu Ser Leu Val Glu Met 245 250 255 Ala Val Gly Arg Tyr
Pro Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu 260 265 270 Leu Leu Phe
Gly Cys His Val Glu Gly Asp Ala Ala Glu Thr Pro Pro 275 280 285 Arg
Pro Arg Thr Pro Gly Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser 290 295
300 Arg Pro Pro Met Ala Ile Phe Glu Leu Leu Asp Tyr Ile Val Asn Glu
305 310 315 320 Pro Pro Pro Lys Leu Pro Ser Gly Val Phe Ser Leu Glu
Phe Gln Asp 325 330 335 Phe Val Asn Lys Cys Leu Ile Lys Asn Pro Ala
Glu Arg Ala Asp Leu 340 345 350 Lys Gln Leu Met Val His Ala Phe Ile
Lys Arg Ser Asp Ala Glu Glu 355 360 365 Val Asp Phe Ala Gly Trp Leu
Cys Ser Thr Ile Gly Leu Asn Gln Pro 370 375 380 Ser Thr Pro Thr His
Ala Ala Ser Ile 385 390 691182DNAOryctolagus cuniculus 69atgccaaaga
agaagcccac ccccatccag ctgaatcctg cccctgacgg ctcggcggtg 60aatggtacca
gctcggcgga gaccaacctg gaggccttgc agaagaagct ggaggagctg
120gagcttgacg agcagcagcg gaagcgcctg gaggccttcc tcacccagaa
gcagaaagtg 180ggagagctga aggacgatga cttcgagaag atcagtgagc
tgggagccgg caacggcggc 240gtggtgttca aggtctccca caagcccagt
ggcctggtga tggccagaaa gcttattcac 300ctggagatca aacctgctat
ccggaaccag atcataaggg agctgcaggt tctgcacgag 360tgcaactccc
cgtacatcgt gggcttctac ggggcattct acagcgatgg cgagatcagc
420atctgcatgg agcacatgga cgggggttcc ttggatcaag tcctgaagaa
agctggacgg 480attcccgagc aaattttggg gaaagttagc attgctgtga
tcaagggcct gacgtatctg 540agggagaagc acaagatcat gcacagagat
gtgaagccct ccaacatcct ggtcaactcc 600cgcggggaga tcaagctctg
tgacttcggg gtcagtgggc agctcatcga ctccatggcc 660aactccttcg
tgggcaccag gtcttatatg tcgcccgaga gactccaggg gacacactac
720tctgtgcagt cggacatctg gagcatgggg ctgtccctgg tggagatggc
ggtggggcgg 780taccccatcc cgccccccga cgccaaggag ctggagctga
tgtttgggtg ccaggtggag 840ggcgatgcgg ccgagactcc gcccaggccc
aggacccctg ggcggcccct cagctcgtat 900ggaatggata gccggcctcc
catggcgatt tttgagctgc tggattacat cgtcaatgag 960cctcctccga
aactccccag cgcagtcttc agcctggagt ttcaagattt tgtgaataaa
1020tgcttaataa aaaaccccgc cgagagagca gacttgaagc agctcatggt
tcatgctttt 1080atcaagaggt ctgatgccga ggaggtggat tttgctggtt
ggctgtgctc caccatcggc 1140cttaaccagc ccagcacgcc gacgcacgcg
gccggtgtgt ga 118270393PRTOryctolagus cuniculus 70Met Pro Lys Lys
Lys Pro Thr Pro Ile Gln Leu Asn Pro Ala Pro Asp 1 5 10 15 Gly Ser
Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu Ala 20 25 30
Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys 35
40 45 Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys Val Gly Glu Leu
Lys 50 55 60 Asp Asp Asp Phe Glu Lys Ile Ser Glu Leu Gly Ala Gly
Asn Gly Gly 65 70 75 80 Val Val Phe Lys Val Ser His Lys Pro Ser Gly
Leu Val Met Ala Arg 85 90 95 Lys Leu Ile His Leu Glu Ile Lys Pro
Ala Ile Arg Asn Gln Ile Ile 100 105 110 Arg Glu Leu Gln Val Leu His
Glu Cys Asn Ser Pro Tyr Ile Val Gly 115 120 125 Phe Tyr Gly Ala Phe
Tyr Ser Asp Gly Glu Ile Ser Ile Cys Met Glu 130 135 140 His Met Asp
Gly Gly Ser Leu Asp Gln Val Leu Lys Lys Ala Gly Arg 145 150 155 160
Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile Ala Val Ile Lys Gly 165
170 175 Leu Thr Tyr Leu Arg Glu Lys His Lys Ile Met His Arg Asp Val
Lys 180 185 190 Pro Ser Asn Ile Leu Val Asn Ser Arg Gly Glu Ile Lys
Leu Cys Asp 195 200 205 Phe Gly Val Ser Gly Gln Leu Ile Asp Ser Met
Ala Asn Ser Phe Val 210 215 220 Gly Thr Arg Ser Tyr Met Ser Pro Glu
Arg Leu Gln Gly Thr His Tyr 225 230 235 240 Ser Val Gln Ser Asp Ile
Trp Ser Met Gly Leu Ser Leu Val Glu Met 245 250 255 Ala Val Gly Arg
Tyr Pro Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu 260 265 270 Leu Met
Phe Gly Cys Gln Val Glu Gly Asp Ala Ala Glu Thr Pro Pro 275 280 285
Arg Pro Arg Thr Pro Gly Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser 290
295 300 Arg Pro Pro Met Ala Ile Phe Glu Leu Leu Asp Tyr Ile Val Asn
Glu 305 310 315 320 Pro Pro Pro Lys Leu Pro Ser Ala Val Phe Ser Leu
Glu Phe Gln Asp 325 330 335 Phe Val Asn Lys Cys Leu Ile Lys Asn Pro
Ala Glu Arg Ala Asp Leu 340 345 350 Lys Gln Leu Met Val His Ala Phe
Ile Lys Arg Ser Asp Ala Glu Glu 355 360 365 Val Asp Phe Ala Gly Trp
Leu Cys Ser Thr Ile Gly Leu Asn Gln Pro 370 375 380 Ser Thr Pro Thr
His Ala Ala Gly Val 385 390 711872DNACavia porcellus 71cgtgtcttcg
tcgggaccgc cctcctcctt gagtcctccc cccaccggga cggccgagtg 60gagaggccgg
acgaaggcgg cggccccggc ggcggctttt cctcggcttc gctgtgcagc
120gtgcgcggcg aggttgaccg cccgcgagcg cccgtgactg agggaacagg
gagagagctc 180gggcggccga gcgcgcagcc ctccgtgggc attgccgcct
ggacctcccg gaaggcaccc 240cgggccgcgg ccgccacccg tcccgccctc
gttcggagct gagacgccgt cgccgcgcaa 300gatgcccaag aagaagccga
cgcccatcca gctgaacccg gcccccgacg gctcggcggt 360gaacgggacc
agctcggccg agaccaacct agaggctttg cagaagaagc tggaggagct
420ggagctggat gagcagcagc ggaagcgcct cgaagctttc ctgacacaga
agcagaaggt 480gggcgagctg aaggacgatg actttgagaa gatcagtgag
ctgggtgccg gcaatggcgg 540tgtggtgttc aaggtctccc acaagccatc
tggcctggtc atggcccgaa agcttatcca 600cctggagatc aagccagcca
tccgcaatca gatcatccgt gagctgcagg ttctgcacga 660gtgcaactcg
ccctacattg tgggcttcta tggggccttc tacagtgatg gcgagatcag
720catctgcatg gagcacatgg atggaggttc cttggatcaa gtcctgaaga
aagctggaag 780aattcctgag caaattttag gaaaagttag cattgctgtg
atcaaaggcc tgacatacct 840gagggagaag cacaagatta tgcacagaga
tgtcaagccc tccaacatcc tggtcaactc 900ccgcggggag atcaagctct
gtgactttgg ggtcagcggg cagctcatcg attccatggc 960caactccttc
gtgggcaccc ggtcctacat gtcgccagag agactgcagg gcacacacta
1020ctcagtgcag tcggacatct ggagcatggg actgtcactg gtggagatgg
cggttgggag 1080gtaccccatc ccccctccag atgccaagga gctggagctg
gtgttcgggt gccaggtgga 1140aggagatgca gctgagatgc cgcccaggcc
caggaccccc ggaagacccc tgagctcata 1200tggaatggac agccggcctc
ccatggcgat tttcgagctg ttggattaca tagtcaacga 1260gccacctccc
aaactgccca gtggagtctt cagtctggaa ttccaggact ttgtaaataa
1320atgcttaata aagaaccctg cggagagagc agacttgaag cagctcatgg
ttcatgcctt 1380catcaagcgc tctgatgctg aggaggtgga cttcgcaggt
tggctctgtg ccaccatcgg 1440ccttaaccag cccagtaccc cgacccacgt
ggccagcatc tgagctgcgg cccggcccag 1500acgtgctctg ccagcagccg
ctatgctctg gcctctccct cgcttctctt cagacgtgcg 1560tttcacctcc
gaccagggtg cagacacagc atgtgccaag ctgtatttgt gttccttttc
1620agtctttatt gccaccgtgt cacccgagtg gatttgcttt gtgcttaggg
ctgtttgtgc 1680tgatgatcac acacacgctg agctgaacag tgacacttgg
tgatgtggtt gtcactgttc 1740tcactccatg tggctggcct gttgcctcca
gtgtctccag acttggggat gtctggtggc 1800acttcccctg ccagggcatc
tcctcagcag agagggaggc ctctgggccc ttgtccttgg 1860cagtgcaagt ga
187272393PRTCavia porcellus 72Met Pro Lys Lys Lys Pro Thr Pro Ile
Gln Leu Asn Pro Ala Pro Asp 1 5 10 15 Gly Ser Ala Val Asn Gly Thr
Ser Ser Ala Glu Thr Asn Leu Glu Ala 20 25 30 Leu Gln Lys Lys Leu
Glu Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys 35 40 45 Arg Leu Glu
Ala Phe Leu Thr Gln Lys Gln Lys Val Gly Glu Leu Lys 50 55 60 Asp
Asp Asp Phe Glu Lys Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly 65 70
75 80 Val Val Phe Lys Val Ser His Lys Pro Ser Gly Leu Val Met Ala
Arg 85 90 95 Lys Leu Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn
Gln Ile Ile 100 105 110 Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser
Pro Tyr Ile Val Gly 115 120 125 Phe Tyr Gly Ala Phe Tyr Ser Asp Gly
Glu Ile Ser Ile Cys Met Glu 130 135 140 His Met Asp Gly Gly Ser Leu
Asp Gln Val Leu Lys Lys Ala Gly Arg 145 150 155 160 Ile Pro Glu Gln
Ile Leu Gly Lys Val Ser Ile Ala Val Ile Lys Gly 165 170 175 Leu Thr
Tyr Leu Arg Glu Lys His Lys Ile Met His Arg Asp Val Lys 180 185 190
Pro Ser Asn Ile Leu Val Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp 195
200 205 Phe Gly Val Ser Gly Gln Leu Ile Asp Ser Met Ala Asn Ser Phe
Val 210 215 220 Gly Thr Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly
Thr His Tyr 225 230 235 240 Ser Val Gln Ser Asp Ile Trp Ser Met Gly
Leu Ser Leu Val Glu Met 245 250 255 Ala Val Gly Arg Tyr Pro Ile Pro
Pro Pro Asp Ala Lys Glu Leu Glu 260 265 270 Leu Val Phe Gly Cys Gln
Val Glu Gly Asp Ala Ala Glu Met Pro Pro 275 280 285 Arg Pro Arg Thr
Pro Gly Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser 290 295 300 Arg Pro
Pro Met Ala Ile Phe Glu Leu Leu Asp Tyr Ile Val Asn Glu 305 310 315
320 Pro Pro Pro Lys Leu Pro Ser Gly Val Phe Ser Leu Glu Phe Gln Asp
325 330 335 Phe Val Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg Ala
Asp Leu 340 345 350 Lys Gln Leu Met Val His Ala Phe Ile Lys Arg Ser
Asp Ala Glu Glu 355 360 365 Val Asp Phe Ala Gly Trp Leu Cys Ala Thr
Ile Gly Leu Asn Gln Pro 370 375 380 Ser Thr Pro Thr His Val Ala Ser
Ile 385 390 731257DNACanis familiaris 73ggagagcgag acacgggccg
ctctccgctc ggagccggac gcgccttccc gcgtccaaaa 60tgcccaagaa gaagccgacg
cccatccagc tgaacccggc ccccgacggc tcggcggtga 120acgggaccag
ctcggcggag accaacctgg aggccttgca gaagaagctg gaggagctgg
180agcttgatga gcagcagcgg aagcgccttg aggcctttct cacccagaag
cagaaggtcg 240gggaactgaa ggatgacgac ttcgagaaga tcagtgagct
gggtgctggc aacggtggcg 300tggtgttcaa ggtctcccac aagccgtccg
gcctagtcat ggccagaaag ctaattcacc 360tggagatcaa acctgcaatc
cggaaccaga tcataaggga gctacaggtt ctacatgagt 420gcaactcccc
gtacatcgtg ggcttctatg gtgcattcta cagcgatggc gagatcagta
480tctgcatgga gcacatggat gggggttcct tggatcaagt cctgaagaaa
gctggaagaa 540ttcctgaaca aattctagga aaagttagca tcgctgtaat
aaaaggtctg acatacctga 600gagagaagca caagattatg cacagagatg
tcaagccttc caacatcctc gtgaactccc 660gtggggagat caagctctgt
gactttgggg tcagcgggca gctcattgac tccatggcca 720actccttcgt
gggcacaagg tcctacatgt cgccagaaag actccagggg actcattact
780ccgtgcagtc ggacatctgg agcatggggc tctctctggt ggagatggca
gttgggaggt 840atcccatccc tcctccggat gccaaggagc tggagctgat
gtttgggtgc caagtggagg 900gagacgtggc tgagacccca cccagaccaa
ggaccccggg aagacccctt agctcttatg 960gaatggacag ccgaccgccc
atggcaattt ttgagctgtt ggattacata gtcaacgagc 1020cccctccaaa
actgcccagt ggagtattca gtctggaatt tcaagatttt gtgaataaat
1080gcttaataaa aaacccagca gagagagcag atctgaagca actcatggtt
catgccttca 1140tcaagagatc tgacggtgaa gaagtggatt ttgcaggttg
gctctgctcc ccccattggc 1200cttaaccagc ccagcacgcc gacccacgca
gctggcgtct aactcgagtc tagagat 125774381PRTCanis familiaris 74Met
Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro Ala Pro Asp 1 5 10
15 Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu Ala
20 25 30 Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln Gln
Arg Lys 35 40 45 Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys Val
Gly Glu Leu Lys 50 55 60 Asp Asp Asp Phe Glu Lys Ile Ser Glu Leu
Gly Ala Gly Asn Gly Gly 65 70 75 80 Val Val Phe Lys Val Ser His Lys
Pro Ser Gly Leu Val Met Ala Arg 85 90 95 Lys Leu Ile His Leu Glu
Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile 100 105
110 Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr Ile Val Gly
115 120 125 Phe Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile Cys
Met Glu 130 135 140 His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys
Lys Ala Gly Arg 145 150 155 160 Ile Pro Glu Gln Ile Leu Gly Lys Val
Ser Ile Ala Val Ile Lys Gly 165 170 175 Leu Thr Tyr Leu Arg Glu Lys
His Lys Ile Met His Arg Asp Val Lys 180 185 190 Pro Ser Asn Ile Leu
Val Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp 195 200 205 Phe Gly Val
Ser Gly Gln Leu Ile Asp Ser Met Ala Asn Ser Phe Val 210 215 220 Gly
Thr Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly Thr His Tyr 225 230
235 240 Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu Val Glu
Met 245 250 255 Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys
Glu Leu Glu 260 265 270 Leu Met Phe Gly Cys Gln Val Glu Gly Asp Val
Ala Glu Thr Pro Pro 275 280 285 Arg Pro Arg Thr Pro Gly Arg Pro Leu
Ser Ser Tyr Gly Met Asp Ser 290 295 300 Arg Pro Pro Met Ala Ile Phe
Glu Leu Leu Asp Tyr Ile Val Asn Glu 305 310 315 320 Pro Pro Pro Lys
Leu Pro Ser Gly Val Phe Ser Leu Glu Phe Gln Asp 325 330 335 Phe Val
Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu 340 345 350
Lys Gln Leu Met Val His Ala Phe Ile Lys Arg Ser Asp Gly Glu Glu 355
360 365 Val Asp Phe Ala Gly Trp Leu Cys Ser Pro His Trp Pro 370 375
380 752095DNAFelis catus 75agccggcaag gagttgagcg tgcggggtgc
ataggcgcgg gtcgtgggag atgaagctgg 60agaggaccaa cctggaggcc ttgcagaaga
agctggagga gctggagctc gatgagcagc 120aacggaagcg cctggaggcc
tttcttaccc agaagcagaa ggtcggggaa ttgaaggatg 180acgacttcga
gaagatcagc gagctgggcg ctggcaacgg tggtgtggtg ttcaaggtct
240cccataagcc gtctggcctg gtcatggcca gaaagctaat tcacctggag
atcaaacctg 300caatccggaa ccagatcata agggagctgc aggttctaca
tgagtgcaac tccccataca 360tcgtgggctt ctatggcgcg ttctacagcg
acggcgagat cagtatctgt atggagcaca 420tggatggggg ttccttggat
caagtcctga agaaagctgg aagaattcct gaacaaattt 480taggaaaagt
tagcattgct gtaataaaag gtctgacata cctgagggag aagcacaaga
540ttatgcacag agatgtcaag ccttccaaca tcctagtgaa ctctcgtggg
gagatcaagc 600tctgtgactt tggggtcagc gggcagctca tcgactccat
ggccaactcc ttcgtgggca 660caaggtccta catgtcgcca gaaagactcc
aggggactca ttactccgtg cagtcggaca 720tctggagcat ggggctatct
ctggttgaga tggcagtcgg gaggtatccc atccctcctc 780ccgatgccaa
ggagctggag ctgatgtttg ggtgccaagt ggagggagat gcggctgaga
840cgccacccag gccgaggacc cccggaaggc ccctcagctc gtatggaatg
gacagccgac 900ctcccatggc aatttttgag ttgttggatt acatagtcaa
cgagcctcct ccaaagctgc 960ccagtggagt attcagtctg gaatttcaag
attttgtgaa taaatgcctc ataaaaaacc 1020cagcagagag agcagatctg
aaacaactca tggttcatgc ctttatcaag agatctgatg 1080gtgaggaagt
ggattttgca ggttggctct gctccaccat cggccttaac cagcccagca
1140caccgaccca cgcggccggc gtctaagtat ctgggaagca gcaaagagcg
agtcccctgc 1200ccagtggtgt gccattgtcg ctttcaggcc tctttgccat
gcctgtctcc gttcagacgt 1260gcatttcgcc tacgacaaag gatgaagaac
acagcatgtg ccaaaattct atttgtgtct 1320tttttaatat tactgtcatt
tattctgtta tttccctaag tggattggct ttgtgcttgg 1380ggctattttt
gtgtatgttg atccaaacat gcgcaacgtt cagttacagt gaaaccttgg
1440tgactgtggg tagtcattct tactgaaaat tgcactgctc ttcccccacc
gtgactggct 1500agctgcctgt agttttggga ttcttttgac acttggtggt
actgcattct tgccgggcgc 1560accttccttc tgttggggta ggagccttgt
aagatccttc acaggcactg catgtgaagc 1620atgctttgct gctatgaaaa
agaacatcag aaagtataga tcttgttatt ttattatatt 1680tttgcttttg
gtgtagaatg aagcaatttc tgtcaaaatc tagccagagc ccttcactgc
1740cacgatagct ggggcttcac cagtctgtct actgtgatga tttgtagact
tctggttgta 1800tttctatatt tattttaaaa tatattatgt gggatattta
gtggtatgtg tctctttaag 1860tttgaattag tgtttctaaa atgatggtta
ctttgaatgt tacaaatgga tcaaggcatt 1920aaaatgtatg agatttatct
ttccccaaat ccaagtaccg atgctattgt aaacaacagt 1980gtgtatagtg
cctaagaatt gtatgaaaat ccttttaacc atttcaaccc agatgtttaa
2040caaatctaat ctcttattct aataaatata ctatcaagtt aaaaggatga aaaaa
209576371PRTFelis catus 76Met Lys Leu Glu Arg Thr Asn Leu Glu Ala
Leu Gln Lys Lys Leu Glu 1 5 10 15 Glu Leu Glu Leu Asp Glu Gln Gln
Arg Lys Arg Leu Glu Ala Phe Leu 20 25 30 Thr Gln Lys Gln Lys Val
Gly Glu Leu Lys Asp Asp Asp Phe Glu Lys 35 40 45 Ile Ser Glu Leu
Gly Ala Gly Asn Gly Gly Val Val Phe Lys Val Ser 50 55 60 His Lys
Pro Ser Gly Leu Val Met Ala Arg Lys Leu Ile His Leu Glu 65 70 75 80
Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile Arg Glu Leu Gln Val Leu 85
90 95 His Glu Cys Asn Ser Pro Tyr Ile Val Gly Phe Tyr Gly Ala Phe
Tyr 100 105 110 Ser Asp Gly Glu Ile Ser Ile Cys Met Glu His Met Asp
Gly Gly Ser 115 120 125 Leu Asp Gln Val Leu Lys Lys Ala Gly Arg Ile
Pro Glu Gln Ile Leu 130 135 140 Gly Lys Val Ser Ile Ala Val Ile Lys
Gly Leu Thr Tyr Leu Arg Glu 145 150 155 160 Lys His Lys Ile Met His
Arg Asp Val Lys Pro Ser Asn Ile Leu Val 165 170 175 Asn Ser Arg Gly
Glu Ile Lys Leu Cys Asp Phe Gly Val Ser Gly Gln 180 185 190 Leu Ile
Asp Ser Met Ala Asn Ser Phe Val Gly Thr Arg Ser Tyr Met 195 200 205
Ser Pro Glu Arg Leu Gln Gly Thr His Tyr Ser Val Gln Ser Asp Ile 210
215 220 Trp Ser Met Gly Leu Ser Leu Val Glu Met Ala Val Gly Arg Tyr
Pro 225 230 235 240 Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu Leu Met
Phe Gly Cys Gln 245 250 255 Val Glu Gly Asp Ala Ala Glu Thr Pro Pro
Arg Pro Arg Thr Pro Gly 260 265 270 Arg Pro Leu Ser Ser Tyr Gly Met
Asp Ser Arg Pro Pro Met Ala Ile 275 280 285 Phe Glu Leu Leu Asp Tyr
Ile Val Asn Glu Pro Pro Pro Lys Leu Pro 290 295 300 Ser Gly Val Phe
Ser Leu Glu Phe Gln Asp Phe Val Asn Lys Cys Leu 305 310 315 320 Ile
Lys Asn Pro Ala Glu Arg Ala Asp Leu Lys Gln Leu Met Val His 325 330
335 Ala Phe Ile Lys Arg Ser Asp Gly Glu Glu Val Asp Phe Ala Gly Trp
340 345 350 Leu Cys Ser Thr Ile Gly Leu Asn Gln Pro Ser Thr Pro Thr
His Ala 355 360 365 Ala Gly Val 370 772405DNABos taurus
77ccggttgact gagggagagt gggagggaat cccgggctgc cgagctgcgc cggcggggaa
60gcccttcggt tccctgtgca ctgagcaagt gggccggggg gttcccccag accgccactg
120gccctttggc cctgacggga ccgcgcaggg cgcgcccccc gaaggcagcc
ttcgggcttg 180cggcccagac ttggccccgc gaggccggcg cggggcagct
cagagggagg aagctagagg 240ggccgccctc agagttggga gcgcctttcc
tgggtccaaa atgcccaaga agaagccgac 300gcccatccag ctgaacccgg
ccccggacgg ctccgcggtt aacgggacca gctcggcgga 360gaccaacctg
gaggccttgc agaagaagct ggaggagctg gagctcgatg aacagcagcg
420gaagcgcctc gaggcctttc tgacccagaa gcagaaggtg ggggaactga
aggatgatga 480ctttgagaag atcagtgagc tgggtgccgg caatggaggt
gtggtgttca aggtctccca 540caagccgtcc ggacttgtta tggccagaaa
gctaattcac ctggagatca aacctgccat 600ccggaaccag atcataaggg
agctgcaggt tctccatgag tgcaactcgc cttatatcgt 660gggcttctac
ggggcgttct acagcgacgg cgagatcagc atctgcatgg agcacatgga
720tgggggttcc ttggatcaag ttctgaagaa agctggaaga attcctgaac
aaattttagg 780aaaagttagc attgctgtaa taaaaggcct gacatacctg
agggagaagc acaagattat 840gcacagagat gtcaagccgt ccaacatcct
agtgaacagc cgtggagaga tcaagctctg 900tgactttggg gtcagcgggc
agctcatcga ctccatggcc aactccttcg tgggcaccag 960gtcctacatg
tcgccagagc gactccaggg gacccattac tccgtgcagt cggacatctg
1020gagcatgggg ctctctctgg ttgagatggc tgtcgggagg tatcccatcc
ctcctccaga 1080tgccaaggag ctggagctga tgtttgggtg ccaggtggag
ggagatgcgg ctgagacccc 1140gcccaggcca aggacccccg ggaggcccct
cagctcttat ggaatggaca gccgacctcc 1200aatggcaatt tttgagttgt
tggattacat agtcaatgag cctcctccaa aactgcccag 1260tggagtattc
agtctggaat ttcaagattt tgtgaataaa tgcttaataa aaaaccccgc
1320agagagagca gatttgaagc aactcatggt tcatgctttt atcaagagat
ctgatgctga 1380ggaagtggat tttgcaggtt ggctctgctc caccatcggc
cttaaccaac ccagcacacc 1440cacccatgcg gctggcgtct aagtggttgg
gaagcagcag tccctgccca agggcatgca 1500ctgttgcttc cgggcagcct
tcccatgcct gtctctgttc agacgtgcat ttcacctatg 1560acaaaggatg
aagaacacag catgtgccaa aattctattt gtgtcatttt caatattatc
1620atctttactc ttattactat tgttattccc ctaagtggat tggctttgtg
cttggggcta 1680tttttgtgta tattgatgat gaagacatgt gcaatgtaga
attacagtga aactctggtg 1740actgtgggta gtcattctta ctgaaaactg
cactgctttc ccacaccatg aactggctgg 1800tcgcctctat tttcgggatt
ctttgacact tggtggtact tcattcttgc caggcatacc 1860ttctaactga
gtaggaagga gccttgtaag atccttcaca ggcagtgcat gtgaagcatg
1920ctttgctgct ataaaaatga gcatcagaaa gtgtgtatca tgttatttta
ttatgttctt 1980gcttttggtg tagaattcag caaattttca tcaaaatcta
gccagagccc ttcactgcca 2040tgatagctgg ggcttcacca gtctgtctac
tgtgatgatt tgtagacttc tggttgtatt 2100tctgtattta tttttaaatc
taccgtgtgg atatttagtg ctatgtctct ttaagtttgg 2160attagtgttt
ctaaaatggt ggagttgctc tgaatgttac aaatggatca aggcattaaa
2220atgaatgaga tctacctttc accaagtact gatgctattg taaacaacag
tgtgtatagt 2280gcctaacaac tgtatgaaaa tccttttacc attttaatcc
agatgtttaa caagcctaat 2340ctcttactct aataaatata ctatcaaatt
caaaggaaaa aaaaaaaaaa aaaaaaaaaa 2400aaaaa 240578393PRTBos taurus
78Met Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro Ala Pro Asp 1
5 10 15 Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu
Ala 20 25 30 Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln
Gln Arg Lys 35 40 45 Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys
Val Gly Glu Leu Lys 50 55 60 Asp Asp Asp Phe Glu Lys Ile Ser Glu
Leu Gly Ala Gly Asn Gly Gly 65 70 75 80 Val Val Phe Lys Val Ser His
Lys Pro Ser Gly Leu Val Met Ala Arg 85 90 95 Lys Leu Ile His Leu
Glu Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile 100 105 110 Arg Glu Leu
Gln Val Leu His Glu Cys Asn Ser Pro Tyr Ile Val Gly 115 120 125 Phe
Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile Cys Met Glu 130 135
140 His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys Lys Ala Gly Arg
145 150 155 160 Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile Ala Val
Ile Lys Gly 165 170 175 Leu Thr Tyr Leu Arg Glu Lys His Lys Ile Met
His Arg Asp Val Lys 180 185 190 Pro Ser Asn Ile Leu Val Asn Ser Arg
Gly Glu Ile Lys Leu Cys Asp 195 200 205 Phe Gly Val Ser Gly Gln Leu
Ile Asp Ser Met Ala Asn Ser Phe Val 210 215 220 Gly Thr Arg Ser Tyr
Met Ser Pro Glu Arg Leu Gln Gly Thr His Tyr 225 230 235 240 Ser Val
Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu Val Glu Met 245 250 255
Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu 260
265 270 Leu Met Phe Gly Cys Gln Val Glu Gly Asp Ala Ala Glu Thr Pro
Pro 275 280 285 Arg Pro Arg Thr Pro Gly Arg Pro Leu Ser Ser Tyr Gly
Met Asp Ser 290 295 300 Arg Pro Pro Met Ala Ile Phe Glu Leu Leu Asp
Tyr Ile Val Asn Glu 305 310 315 320 Pro Pro Pro Lys Leu Pro Ser Gly
Val Phe Ser Leu Glu Phe Gln Asp 325 330 335 Phe Val Asn Lys Cys Leu
Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu 340 345 350 Lys Gln Leu Met
Val His Ala Phe Ile Lys Arg Ser Asp Ala Glu Glu 355 360 365 Val Asp
Phe Ala Gly Trp Leu Cys Ser Thr Ile Gly Leu Asn Gln Pro 370 375 380
Ser Thr Pro Thr His Ala Ala Gly Val 385 390 792146DNAEquus caballus
79caagtgggaa agcttgggat gcgtggagga gccagctact agtttagtgt gctgggtgcg
60tcggtgcagg tcgcaggaga tgaagccgga gaggaccaac ctggaggcct tgcagaagaa
120gctggaggag ctggagctcg atgaacagca gcgaaagcgc cttgaggcct
ttcttactca 180gaagcagaag gttggggaac tgaaggatga tgactttgag
aagatcagtg agctgggtgc 240tggcaatggt ggtgtggtat tcaaggttgc
ccacaaaccg tctggtttgg tcatggccag 300aaagctaatt cacctggaga
tcaagcctgc aatccggaac cagatcataa gggagctgca 360ggttctacat
gagtgcaact ccccatatat tgtgggcttc tatggcgcat tctacagcga
420tggtgagatc agcatctgca tggagcacat ggatgggggt tccttggatc
aagtcctaaa 480gaaagctgga agaattcctg agcaaatttt aggaaaagtt
agcattgctg taataaaagg 540cctgacgtat ctgagggaga agcacaagat
tatgcacaga gatgtcaagc cctccaacat 600cctagtgaac tcccgtgggg
agatcaagct gtgtgatttt ggggtcagcg ggcagctcat 660cgactccatg
gccaactcct tcgtgggcac aaggtcttac atgtcgccgg aaagactcca
720ggggactcat tattcagtgc agtcggacat ctggagcatg gggctctctc
tggttgagat 780ggcggtcggg aggtatccca tccctcctcc agatgccaag
gagctggagc tgatgtttgg 840gtgccaagtg gagggagatg cggctgagac
tccgcctagg ccaaggaccc ctggaagacc 900cctcagctct tatggaatgg
acagccgacc tcctatggca atttttgagt tactggatta 960catagtcaac
gagcctcctc ccaagctgcc cagtggagta ttcagtctgg aatttcagga
1020ttttgtgaat aaatgcttaa tcaaaaaccc tgcagagaga gcagatttga
agcaactcat 1080ggttcacgct tttatcaaga gatctgatgc cgaggaagtg
gattttgcag gttggctctg 1140ctccaccatt ggccttaacc agcccagcac
accaacccac gcggctggcg tctaagcgtt 1200tgggaagcag caaaaagcga
gccccctgcc gcgtggtgtg ccatgttgct ttcgggcctc 1260cttcccatgc
ctgtctgttc acacgtgcat ttcacctgtg acaaaggatg aagaacacag
1320catgtgccaa aattctattt gtgtcatttt taatagtact gtctttattc
ttattactat 1380tgttattccc ctaagtggat tggctttgtg cttgggacta
ttttgtgtat gttgatgatc 1440aaaacatgcg caatgttgaa ttaccgtgaa
actggtgact gtgggtagtc cttcttattg 1500aaaattgcac tgctcttccc
tccctgtcac tggctggctg cctgtatttc tggggttctt 1560tgacacttgg
tggtacttca ttcttgcagg gcatacctcc tattcgagta ggaaggagcc
1620tttaagatcc ttcacaggca gtgcatgtga agcatgcttt gctgctatga
aaatgagcat 1680cagaaagtgt atatcatgtt attttattat tattatgttt
ttgcttttgg tgtagaattc 1740agcaatttcc atcaagatct agccagagcc
cttcactgcc atgatagctg gggcttcacc 1800agtctgccta ctgtgatgat
ttgtagactt ctggttgtat ttctatattt atttttaaat 1860atactgtgtg
ggatatttag tggtatatgt ctctctaagt ttggagtggt gtttctaaaa
1920tggagttact ttgaatgtta tagatggatc aaggcataaa atgtatgaga
tttatttttc 1980cccaaatcca aatactgatg ctattgtaaa caacaaacag
tgtgtatagt gcctaaaaat 2040tgtatgaaag tccttttaac cattttaatc
cagatgttta acaaatctaa tctcttattc 2100taataaatat actatcaagt
taaacggaca aaagatttct actttc 214680371PRTEquus caballus 80Met Lys
Pro Glu Arg Thr Asn Leu Glu Ala Leu Gln Lys Lys Leu Glu 1 5 10 15
Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys Arg Leu Glu Ala Phe Leu 20
25 30 Thr Gln Lys Gln Lys Val Gly Glu Leu Lys Asp Asp Asp Phe Glu
Lys 35 40 45 Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly Val Val Phe
Lys Val Ala 50 55 60 His Lys Pro Ser Gly Leu Val Met Ala Arg Lys
Leu Ile His Leu Glu 65 70 75 80 Ile Lys Pro Ala Ile Arg Asn Gln Ile
Ile Arg Glu Leu Gln Val Leu 85 90 95 His Glu Cys Asn Ser Pro Tyr
Ile Val Gly Phe Tyr Gly Ala Phe Tyr 100 105 110 Ser Asp Gly Glu Ile
Ser Ile Cys Met Glu His Met Asp Gly Gly Ser 115 120 125 Leu Asp Gln
Val Leu Lys Lys Ala Gly Arg Ile Pro Glu Gln Ile Leu 130 135 140 Gly
Lys Val Ser Ile Ala Val Ile Lys Gly Leu Thr Tyr Leu Arg Glu 145 150
155 160 Lys His Lys Ile Met His Arg Asp Val Lys Pro Ser Asn Ile Leu
Val 165 170 175 Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp Phe Gly Val
Ser Gly Gln 180 185 190 Leu Ile Asp Ser Met Ala Asn Ser Phe Val Gly
Thr Arg Ser Tyr Met 195 200 205 Ser Pro Glu Arg Leu Gln Gly Thr His
Tyr Ser Val Gln Ser Asp Ile 210 215 220
Trp Ser Met Gly Leu Ser Leu Val Glu Met Ala Val Gly Arg Tyr Pro 225
230 235 240 Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu Leu Met Phe Gly
Cys Gln 245 250 255 Val Glu Gly Asp Ala Ala Glu Thr Pro Pro Arg Pro
Arg Thr Pro Gly 260 265 270 Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser
Arg Pro Pro Met Ala Ile 275 280 285 Phe Glu Leu Leu Asp Tyr Ile Val
Asn Glu Pro Pro Pro Lys Leu Pro 290 295 300 Ser Gly Val Phe Ser Leu
Glu Phe Gln Asp Phe Val Asn Lys Cys Leu 305 310 315 320 Ile Lys Asn
Pro Ala Glu Arg Ala Asp Leu Lys Gln Leu Met Val His 325 330 335 Ala
Phe Ile Lys Arg Ser Asp Ala Glu Glu Val Asp Phe Ala Gly Trp 340 345
350 Leu Cys Ser Thr Ile Gly Leu Asn Gln Pro Ser Thr Pro Thr His Ala
355 360 365 Ala Gly Val 370 812274DNAGallus gallus 81ctaaccaggc
gggagctgtc ggtgcggagc tcggtgtcgc tccgccgggc aggccgggtc 60gacggccgcg
ctgtgccgga gcggcagcgt cgcgggctcg gctccttctc ggggaggcgg
120ccgcgcgctg ctccggcgct gaggggcggc cccgaagttt gcttcgcgtc
gggaagtccg 180tcggacctgg ccgaagtggg gccgcggccg ctccgtccgt
cacgctctgc gctggccggg 240gggcaacatg cccaagaaga agccagggcc
gatccagctc aaccccgctc cggatggctc 300cgccgtcaac gggaccagct
ctgccgagac aaacttggaa gctcttcaga agaagctgga 360agagctagag
ctggatgaac agcaaaggaa gcgccttgaa gctttcctta cccagaaaca
420aaaagttggg gagctgaagg atgatgactt cgagaagatc agtgagctgg
gagcagggaa 480tggcggggtg gtgttcaaag tatctcacaa gccttctggc
ctcatcatgg caagaaaatt 540gattcatcta gaaatcaagc cagctattcg
aaaccagatc atccgtgagc tgcaggttct 600acatgagtgc aattcaccat
acatagtggg cttttatgga gctttttaca gtgatgggga 660aatcagcatt
tgcatggaac acatggatgg tggctcattg gatcaagtgc tgaaaaaggc
720tggaagaatt ccagagcaga tactgggcaa agtcagcatt gcagtaataa
aaggactcac 780atatctgaga gaaaagcata aaataatgca cagagatgtt
aaaccatcta acatcttggt 840aaactctaga ggtgagatca agctttgtga
ttttggtgtc agtggacaac tgatagattc 900tatggcaaac tcatttgttg
gcacgcgctc ctacatgtct ccggaaagac tgcagggaac 960tcattattca
gtgcagtcag atatatggag tatggggctg tctctggtag aaatggccat
1020tggcagatac ccgattcctc ctcctgactc taaggagctc gagttgatgt
ttggctgccc 1080ggtagaggga gattctccag tcacagagac ctcacccagg
caaagaacac ctggtcgacc 1140aatgagctcc tatggaccag acagcagacc
cccgatggca atctttgaac ttctggatta 1200catcgtcaat gagccacctc
caaaactgcc caatggtgtc tttggttctg aatttcaaga 1260ttttgttaac
aaatgtttaa ttaaaaatcc tgctgagaga gctgatttga agcagctgat
1320gattcacgct ttcattaaga gatctgaagc agaggaggtg gattttgcag
gatggctttg 1380ctcaaccata ggccttaacc aaccgagtac acccacgcat
gctgctggag tctgaatgtg 1440gaagagcaaa tcctgtcccg tacatctgtt
aacagcgcta ctttggtcct atttcctaag 1500cttgtacctg ttcaaacatg
tatttcacct cttaaggaag aatgtcttta tagcatgtgc 1560caaattgttt
tcaattttgt catcaactaa ttggtattgt actgggttac atttgtttgc
1620tgaccaaaat gtaaaatgtt taagttacag tgcttgctga ttttaagtga
ttatggaatt 1680atggatattc tttcttaatg aaaatatcac tgggggggaa
tttacccctg gattgtttga 1740actttatcaa gactctttgt aaactgttgg
tacttcagtc atgcttacct aatctcccat 1800gcaaaaaaag gggtagggat
gctccaaaac tgtatctgtt gagcatgctt ttgctgctgc 1860caaactgtat
cttggaagtt aggcctaatg gttccaattt ggtgttgtgt agagatcact
1920ctttccaggt aaagaaggta agagctctgc attccttggg atggacaggg
cagtatccta 1980cttgtagact tgttcatatt tctatattta tttttaaaat
gtatcatcat acttggattt 2040agtgatatat gtctttccaa ttgattttta
aaggttagct ctcagaagcg tcctacagaa 2100tcatgacaaa gatctgggct
ttcttttaac cttaagattc atgacagctg tgtttggtgt 2160ctaaaatgta
tgaagatcct ctattgtttt attctctcag atgtttagca atggtttctc
2220ttaataaata tattatcaag taaaaaaaaa aaaaaaataa aaaaaaaaaa aaaa
227482395PRTGallus gallus 82Met Pro Lys Lys Lys Pro Gly Pro Ile Gln
Leu Asn Pro Ala Pro Asp 1 5 10 15 Gly Ser Ala Val Asn Gly Thr Ser
Ser Ala Glu Thr Asn Leu Glu Ala 20 25 30 Leu Gln Lys Lys Leu Glu
Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys 35 40 45 Arg Leu Glu Ala
Phe Leu Thr Gln Lys Gln Lys Val Gly Glu Leu Lys 50 55 60 Asp Asp
Asp Phe Glu Lys Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly 65 70 75 80
Val Val Phe Lys Val Ser His Lys Pro Ser Gly Leu Ile Met Ala Arg 85
90 95 Lys Leu Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn Gln Ile
Ile 100 105 110 Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr
Ile Val Gly 115 120 125 Phe Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile
Ser Ile Cys Met Glu 130 135 140 His Met Asp Gly Gly Ser Leu Asp Gln
Val Leu Lys Lys Ala Gly Arg 145 150 155 160 Ile Pro Glu Gln Ile Leu
Gly Lys Val Ser Ile Ala Val Ile Lys Gly 165 170 175 Leu Thr Tyr Leu
Arg Glu Lys His Lys Ile Met His Arg Asp Val Lys 180 185 190 Pro Ser
Asn Ile Leu Val Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp 195 200 205
Phe Gly Val Ser Gly Gln Leu Ile Asp Ser Met Ala Asn Ser Phe Val 210
215 220 Gly Thr Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly Thr His
Tyr 225 230 235 240 Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser
Leu Val Glu Met 245 250 255 Ala Ile Gly Arg Tyr Pro Ile Pro Pro Pro
Asp Ser Lys Glu Leu Glu 260 265 270 Leu Met Phe Gly Cys Pro Val Glu
Gly Asp Ser Pro Val Thr Glu Thr 275 280 285 Ser Pro Arg Gln Arg Thr
Pro Gly Arg Pro Met Ser Ser Tyr Gly Pro 290 295 300 Asp Ser Arg Pro
Pro Met Ala Ile Phe Glu Leu Leu Asp Tyr Ile Val 305 310 315 320 Asn
Glu Pro Pro Pro Lys Leu Pro Asn Gly Val Phe Gly Ser Glu Phe 325 330
335 Gln Asp Phe Val Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg Ala
340 345 350 Asp Leu Lys Gln Leu Met Ile His Ala Phe Ile Lys Arg Ser
Glu Ala 355 360 365 Glu Glu Val Asp Phe Ala Gly Trp Leu Cys Ser Thr
Ile Gly Leu Asn 370 375 380 Gln Pro Ser Thr Pro Thr His Ala Ala Gly
Val 385 390 395
* * * * *