U.S. patent application number 14/620649 was filed with the patent office on 2015-06-04 for novel ring-substituted n-pyridinyl amides as kinase inhibitors.
The applicant listed for this patent is Novartis AG. Invention is credited to Matthew Burger, Joseph E. Drumm III, Gisele Nishiguchi, Alice Rico, Robert Lowell Simmons, Benjamin Taft, Huw Tanner.
Application Number | 20150150873 14/620649 |
Document ID | / |
Family ID | 48699902 |
Filed Date | 2015-06-04 |
United States Patent
Application |
20150150873 |
Kind Code |
A1 |
Burger; Matthew ; et
al. |
June 4, 2015 |
NOVEL RING-SUBSTITUTED N-PYRIDINYL AMIDES AS KINASE INHIBITORS
Abstract
The present invention provides a compound of formula (A):
##STR00001## as described herein, and pharmaceutically acceptable
salts, enantiomers, rotamers, tautomers, or racemates thereof. Also
provided are methods of treating a disease or condition mediated by
PIM kinase using the compounds of Formula I, and pharmaceutical
compositions comprising such compounds.
Inventors: |
Burger; Matthew; (Albany,
CA) ; Drumm III; Joseph E.; (Walnut Creek, CA)
; Nishiguchi; Gisele; (Albany, CA) ; Rico;
Alice; (Castro Valley, CA) ; Simmons; Robert
Lowell; (San Francisco, CA) ; Taft; Benjamin;
(Oakland, CA) ; Tanner; Huw; (Alameda,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Novartis AG |
Basel |
|
CH |
|
|
Family ID: |
48699902 |
Appl. No.: |
14/620649 |
Filed: |
February 12, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13897927 |
May 20, 2013 |
8987457 |
|
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14620649 |
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61695533 |
Aug 31, 2012 |
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61649645 |
May 21, 2012 |
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Current U.S.
Class: |
514/256 ;
514/278; 514/318; 514/332; 514/333; 514/342 |
Current CPC
Class: |
A61K 31/675 20130101;
C07D 403/14 20130101; A61P 43/00 20180101; C07D 409/14 20130101;
A61K 31/444 20130101; C07D 417/12 20130101; C07F 9/65583 20130101;
C07D 403/12 20130101; C07D 401/12 20130101; A61K 45/06 20130101;
A61P 29/00 20180101; A61P 37/02 20180101; C07D 213/81 20130101;
A61K 31/506 20130101; A61P 35/00 20180101; C07D 407/14 20130101;
A61P 1/04 20180101; A61K 31/4545 20130101; C07D 417/14 20130101;
A61K 31/4439 20130101; A61P 35/02 20180101; C07D 405/14 20130101;
C07D 401/14 20130101; A61P 37/00 20180101 |
International
Class: |
A61K 31/506 20060101
A61K031/506; A61K 31/4545 20060101 A61K031/4545; A61K 31/4439
20060101 A61K031/4439; A61K 31/444 20060101 A61K031/444 |
Claims
1-21. (canceled)
22. A method of treating a disease or condition selected from
carcinoma of the lungs, pancreas, thyroid, ovaries, bladder,
breast, prostate or colon, melanoma, myeloid leukemia, multiple
myeloma, erythro leukemia, villous colon adenoma, and osteosarcoma,
comprising administering to a subject in need thereof a
therapeutically effective amount of a compound selected from:
##STR00421## ##STR00422## ##STR00423## ##STR00424## ##STR00425##
##STR00426## ##STR00427## ##STR00428## ##STR00429## ##STR00430##
##STR00431## ##STR00432## ##STR00433## ##STR00434## ##STR00435##
##STR00436## ##STR00437## ##STR00438## ##STR00439## ##STR00440##
##STR00441## ##STR00442## ##STR00443## ##STR00444## ##STR00445##
##STR00446## ##STR00447## ##STR00448## ##STR00449## ##STR00450##
##STR00451## ##STR00452## ##STR00453## ##STR00454## ##STR00455##
##STR00456## ##STR00457## ##STR00458## ##STR00459## ##STR00460##
##STR00461## ##STR00462## ##STR00463## ##STR00464## ##STR00465##
##STR00466## ##STR00467## ##STR00468## ##STR00469## ##STR00470##
##STR00471## ##STR00472##
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. provisional
application Serial Nos. 61/649,645, filed 21 May 2012, and
61/695,533, filed 31 Aug. 2012, the contents of which are
incorporated herein by reference in their entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to new compounds and their
tautomers and pharmaceutically acceptable salts, esters,
metabolites or prodrugs thereof, compositions of the new compounds
together with pharmaceutically acceptable carriers, and uses of the
new compounds, either alone or in combination with at least one
additional therapeutic agent, in the prophylaxis or treatment of
cancer and other cellular proliferation disorders.
BACKGROUND
[0003] Infection with the Moloney retrovirus and genome integration
in the host cell genome results in development of lymphomas in
mice. Provirus Integration of Moloney Kinase (PIM-Kinase) was
identified as one of the frequent proto-oncogenes capable of being
transcriptionally activated by this retrovirus integration event
(Cuypers H T et al., "Murine leukemia virus-induced T-cell
lymphomagenesis: integration of proviruses in a distinct
chromosomal region," Cell 37(1):141-50 (1984); Selten G, et al.,
"Proviral activation of the putative oncogene Pim-1 in MuLV induced
T-cell lymphomas" EMBO J 4(7):1793-8 (1985)), thus establishing a
correlation between over-expression of this kinase and its
oncogenic potential. Sequence homology analysis demonstrated that
there are three highly homologous Pim-Kinases (Pim1, 2 & 3),
Pim1 being the proto-oncogene originally identified by retrovirus
integration. Furthermore, transgenic mice over-expressing Pim1 or
Pim2 show increased incidence of T-cell lymphomas (Breuer M et al.,
"Very high frequency of lymphoma induction by a chemical carcinogen
in pim-1 transgenic mice" Nature 340(6228):61-3 (1989)), while
over-expression in conjunction with c-myc is associated with
incidence of B-cell lymphomas (Verbeek S et al., "Mice bearing the
E mu-myc and E mu-pim-1 transgenes develop pre-B-cell leukemia
prenatally" Mol Cell Biol 11(2):1176-9 (1991)). Thus, these animal
models establish a strong correlation between Pim over-expression
and oncogenesis in hematopoietic malignancies.
[0004] In addition to these animal models, Pim over-expression has
been reported in many human malignancies. Pim1, 2 & 3
over-expression is frequently observed in hematopoietic
malignancies (Amson R et al., "The human protooncogene product
p33pim is expressed during fetal hematopoiesis and in diverse
leukemias," PNAS USA 86(22):8857-61 (1989); Cohen A M et al.,
"Increased expression of the hPim-2 gene in human chronic
lymphocytic leukemia and non-Hodgkin lymphoma," Leuk Lymph
45(5):951-5 (2004), Huttmann A et al., "Gene expression signatures
separate B-cell chronic lymphocytic leukaemia prognostic subgroups
defined by ZAP-70 and CD38 expression status," Leukemia
20:1774-1782 (2006)) and in prostate cancer (Dhanasekaran S M, et
al., "Delineation of prognostic biomarkers in prostate cancer,"
Nature 412(6849):822-6 (2001); Cibull T L, et al., "Overexpression
of Pim-1 during progression of prostatic adenocarcinoma," J Clin
Pathol 59(3):285-8 (2006)), while over-expression of Pim3 is
frequently observed in hepatocellular carcinoma (Fujii C, et al.,
"Aberrant expression of serine/threonine kinase Pim-3 in
hepatocellular carcinoma development and its role in the
proliferation of human hepatoma cell lines," Int J Cancer
114:209-218 (2005)) and pancreatic cancer (Li Y Y et al., "Pim-3, a
proto-oncogene with serine/threonine kinase activity, is aberrantly
expressed in human pancreatic cancer and phosphorylates bad to
block bad-mediated apoptosis in human pancreatic cancer cell
lines," Cancer Res 66(13):6741-7 (2006)).
[0005] Pim1, 2 & 3 are Serine/Threonine kinases that normally
function in survival and proliferation of hematopoietic cells in
response to growth factors and cytokines Cytokines signaling
through the Jak/Stat pathway leads to activation of transcription
of the Pim genes and synthesis of the proteins. No further
post-translational modifications are required for the Kinase Pim
activity. Thus, signaling downstream is primarily controlled at the
transcriptional/translational and protein turnover level.
Substrates for Pim kinases include regulators of apoptosis such as
the Bc1-2 family member BAD (Aho T et al., "Pim-1 kinase promotes
inactivation of the pro-apoptotic Bad protein by phosphorylating it
on the Ser112 gatekeeper site: FEBS Letters 571: 43-49 (2004)),
cell cycle regulators such as p21.sup.WFA1/CIP1 (Wang Z, et al.,
"Phosphorylation of the cell cycle inhibitor p21Cip1/WAF1 by Pim-1
kinase," Biochem Biophys Acta 1593:45-55 (2002)), CDC25A (1999),
C-TAK (Bachmann M et al., "The Oncogenic Serine/Threonine Kinase
Pim-1 Phosphorylates and Inhibits the Activity of Cdc25C-associated
Kinase 1 (C-TAK1). A novel role for Pim-1 at the G2/M cell cycle
checkpoint," J Biol Chem 179:48319-48328 (2004)) and NuMA
(Bhattacharya N, et al., "Nm-1 associates with protein complexes
necessary for mitosis," Chromosoma 111(2):80-95 (2002)) and the
protein synthesis regulator 4EBP1 (Hammerman P S et al., "Pim and
Akt oncogenes are independent regulators of hematopoietic cell
growth and survival," Blood 105(11):4477-83 (2005)). The effects of
Pim(s) in these regulators are consistent with a role in protection
from apoptosis and promotion of cell proliferation and growth.
Thus, over-expression of Pim(s) in cancer is thought to play a role
in promoting survival and proliferation of cancer cells and,
therefore, their inhibitions should be an effective way of treating
cancers in which they are over-expressed. In fact several reports
indicate that knocking down expression of Pim(s) with siRNA results
in inhibition of proliferation and cell death (Dai J M, et al.,
"Antisense oligodeoxynucleotides targeting the serine/threonine
kinase Pim-2 inhibited proliferation of DU-145 cells," Acta
Pharmacol Sin 26(3):364-8 (2005); Fujii et al. 2005; Li et al.
2006).
[0006] Furthermore, mutational activation of several well known
oncogenes in hematopoietic malignancies is thought to exert its
effects at least in part through Pim(s). For example, targeted
down-regulation of Pim expression impairs survival of hematopoietic
cells transformed by Flt3 and BCR/ABL (Adam et al. 2006). Thus,
inhibitors to Pim1, 2 and 3 would be useful in the treatment of
these malignancies.
[0007] In addition to a potential role in cancer treatment and
myeloproliferative diseases, such inhibitor could be useful to
control expansion of immune cells in other pathologic condition
such as autoimmune diseases, allergic reactions and in organ
transplantation rejection syndromes. This notion is supported by
the findings that differentiation of Th1 Helper T-cells by IL-12
and IFN-.alpha. results in induction of expression of both Pim1 and
Pim2 (Aho T et al., "Expression of human Pim family genes is
selectively up-regulated by cytokines promoting T helper type 1,
but not T helper type 2, cell differentiation," Immunology 116:
82-88 (2005)). Moreover, Pim(s) expression is inhibited in both
cell types by the immunosuppressive TGF-.beta. (Aho et al. 2005).
These results suggest that Pim kinases are involved in the early
differentiation process of Helper T-cells, which coordinate the
immunological responses in autoimmune diseases, allergic reaction
and tissue transplant rejection. Recent reports demonstrate that
Pim kinase inhibitors show activity in animal models of
inflammation and autoimmune diseases. See JE Robinson "Targeting
the Pim Kinase Pathway for Treatment of Autoimmune and Inflammatory
Diseases," for the Second Annual Conference on Anti-Inflammatories:
Small Molecule Approaches," San Diego, Calif. (Conf. April 2011;
Abstract published earlier on-line).
[0008] A continuing need exists for compounds that inhibit the
proliferation of capillaries, inhibit the growth of tumors, treat
cancer, modulate cell cycle arrest, and/or inhibit molecules such
as Pim1, Pim2 and Pim3, and pharmaceutical formulations and
medicaments that contain such compounds. A need also exists for
methods of administering such compounds, pharmaceutical
formulations, and medicaments to patients or subjects in need
thereof. The present invention addresses such needs.
[0009] Earlier patent applications have described compounds that
inhibit Pims and function as anticancer therapeutics, see, e.g.,
WO2012/004217, WO2010/026124, WO 2008/106692 and WO2011/124580, and
as treatment for inflammatory conditions such as Crohn's disease,
inflammatory bowel disease, rheumatoid arthritis, and chronic
inflammatory diseases, see e.g., WO 2008/022164. The present
invention provides compounds that inhibit activity of one or more
Pims, preferably two or more Pims, more preferably Pim1, Pim2 and
Pim3, at nanomolar levels (e.g., IC-50 under 100 nM or under 50 nM)
and exhibit distinctive characteristics that may provide improved
therapeutic effects and pharmacokinetic properties, such as reduced
drug-drug interactions associated with inhibition of cytochrome
oxidases, relative to compounds previously disclosed. Compounds of
the invention contain novel substitution combinations on one or
more rings that provide these distinctive properties and are
suitable for treating Pim-related conditions such as those
described herein.
SUMMARY OF THE INVENTION
[0010] The invention provides compounds of Formula A, having three
substituents on a cyclohexyl or piperidinyl ring that is attached
to an N-pyridinyl amide moiety of Formula (A):
##STR00002##
[0011] wherein:
[0012] groups attached to the ring containing Q that are depicted
inside the ring are all syn to each other, and all groups attached
to that ring that are depicted outside the ring are syn to one
another;
[0013] Q is C or N;
[0014] R.sup.q is H when Q is C, and R.sup.q is absent when Q is
N;
[0015] R.sup.1 is selected from H, C.sub.1-4 alkyl, C.sub.3-6
cycloalkyl, C.sub.4-6 heterocyclyl, --(CR'.sub.2).sub.1-3 OR' and
--OR',
[0016] where each R' is independently H or C.sub.1-4 alkyl,
and each alkyl, cycloalkyl and heterocyclyl is optionally
substituted with up to two groups selected from halo, CN, NH.sub.2,
hydroxy, oxo, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy, and C.sub.1-4
haloalkoxy;
[0017] R.sup.1b, R.sup.1c, and R.sup.1d are each independently
selected from H, halo, OR', R', --(CH.sub.2).sub.1-2OR', and
CONR'.sub.2;
[0018] one of R.sup.2a and R.sup.2b is H, and the other of R.sup.2a
and R.sup.2b is selected from CN, halo, azido, amino, --OR,
--O(CH.sub.2).sub.1-3OR, --NRC(O)R, --NRC(O)OR, --NHSO.sub.2R,
--SO.sub.2R, --OSO.sub.2R, --SR, --S(O)R, --OP(O)R.sub.2, and
1-pyridonyl or 1-triazolyl optionally substituted with up to two
groups selected from halo, C.sub.1-4 alkyl, hydroxy-substituted
C.sub.1-4 alkyl, C.sub.2-4 alkenyl, C.sub.3-6 cycloalkyl, C.sub.1-4
alkoxy, and C.sub.5-6 heteroaryl;
[0019] where each R is independently C.sub.5-6 heteroaryl or
C.sub.1-4 alkyl optionally substituted with up to three groups
selected from cyano, halo, hydroxy, carboxy, C.sub.1-4
alkylsulfonyl, and C.sub.1-4 alkoxy; or R.sup.2a and R.sup.2b taken
together may form a dialkyl ketal or 5-6 membered cyclic ketal,
.dbd.O or .dbd.N--OR'', where R'' is H or C.sub.1-4 alkyl;
[0020] ring A is selected from pyridinyl, pyrimidinyl, pyrazinyl,
and thiazolyl, each having N positioned as shown in Formula (I);
and
[0021] Ring A is optionally substituted with 1 or 2 groups selected
from halo, CN, NH.sub.2, hydroxy, C.sub.1-4 alkyl, C.sub.1-4
haloalkyl, C.sub.1-4 alkoxy, and C.sub.1-4 haloalkoxy; [0022] or a
pharmaceutically acceptable salt thereof.
[0023] In some embodiments of the compounds of Formula A, R1b is H
and R1 is selected from H, C1-4 alkyl, C3-6 cycloalkyl, C4-6
heterocyclyl, --(CR'2)1-3-OR' and --OR',
[0024] where each R' is independently H or C.sub.1-4 alkyl, and
each alkyl, cycloalkyl and heterocyclyl is optionally substituted
with up to two groups selected from halo, CN, NH.sub.2, hydroxy,
oxo, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy, and C.sub.1-4
haloalkoxy;
[0025] one of R.sup.2a and R.sup.2b is H, and the other of R.sup.2a
and R.sup.2b is selected from CN, halo, azido, amino, --OR,
--O(CH.sub.2).sub.1-3OR, --NRC(O)R, --NRC(O)OR, --NHSO.sub.2R,
--SO.sub.2R, --OSO.sub.2R, --SR, --S(O)R, --OP(O)R.sub.2, and
triazolyl optionally substituted with up to two groups selected
from halo, C.sub.1-4 alkyl, hydroxy-substituted C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, and C.sub.5-6 heteroaryl;
[0026] where each R is independently C.sub.1-4 alkyl optionally
substituted with up to three groups selected from cyano, halo,
hydroxy, carboxy, C.sub.1-4 alkylsulfonyl, and C.sub.1-4 alkoxy; or
R.sup.2a and R.sup.2b taken together form a dialkyl ketal or 5-6
membered cyclic ketal, .dbd.O or .dbd.N--OR'', where R'' is H or
C.sub.1-4 alkyl;
[0027] Ring A is selected from pyridinyl, pyrimidinyl, pyrazinyl,
and thiazolyl, each having N positioned as shown in Formula (I);
and
[0028] Ring A is optionally substituted with 1 or 2 groups selected
from halo, CN, NH.sub.2, hydroxy, C.sub.1-4 alkyl, C.sub.1-4
haloalkyl, C.sub.1-4 alkoxy, and C.sub.1-4 haloalkoxy;
[0029] or a pharmaceutically acceptable salt thereof.
[0030] Additional embodiments of these compounds and pharmaceutical
compositions and uses for these compounds and compositions are
described below.
[0031] These compounds are inhibitors of Pim kinases as further
discussed herein. These compounds and their pharmaceutically
acceptable salts, and pharmaceutical compositions containing these
compounds and salts are useful for therapeutic methods such as
treatment of cancers and autoimmune disorders that are caused by or
exacerbated by excessive levels of Pim kinase activity.
DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION
[0032] "PIM inhibitor" is used herein to refer to a compound that
exhibits an IC.sub.50 with respect to PIM Kinase activity of no
more than about 100 .mu.M and more typically not more than about 50
.mu.M, as measured in the PIM depletion assays described herein
below for at least one of Pim1, Pim2 and Pim3. Preferred compounds
have on IC.sub.50 below about 1 micromolar on at least one Pim, and
generally have an IC.sub.50 below 100 nM on each of Pim1, Pim2 and
Pim3.
[0033] The phrase "alkyl" refers to hydrocarbon groups that do not
contain heteroatoms, i.e., they consist of carbon atoms and
hydrogen atoms. Thus the phrase includes straight chain alkyl
groups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl,
octyl, nonyl, decyl, undecyl, dodecyl and the like. The phrase also
includes branched chain isomers of straight chain alkyl groups,
including but not limited to, the following which are provided by
way of example: --CH(CH.sub.3).sub.2,
--CH(CH.sub.3)(CH.sub.2CH.sub.3), --CH(CH.sub.2CH.sub.3).sub.2,
--C(CH.sub.3).sub.3, --C(CH.sub.2CH.sub.3).sub.3,
--CH.sub.2CH(CH.sub.3).sub.2, --CH.sub.2
CH(CH.sub.3)(CH.sub.2CH.sub.3),
--CH.sub.2CH(CH.sub.2CH.sub.3).sub.2, --CH.sub.2C(CH.sub.3).sub.3,
--CH.sub.2C(CH.sub.2CH.sub.3).sub.3,
--CH(CH.sub.3)--CH(CH.sub.3)(CH.sub.2CH.sub.3),
--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2,
--CH.sub.2CH.sub.2CH(CH.sub.3)(CH.sub.2CH.sub.3),
--CH.sub.2CH.sub.2CH(CH.sub.2CH.sub.3).sub.2,
--CH.sub.2CH.sub.2C(CH.sub.3).sub.3,
--CH.sub.2CH.sub.2C(CH.sub.2CH.sub.3).sub.3,
--CH(CH.sub.3)CH.sub.2-- CH(CH.sub.3).sub.2,
--CH(CH.sub.3)CH(CH.sub.3)CH(CH.sub.3).sub.2,
--CH(CH.sub.2CH.sub.3)CH(CH.sub.3)CH(CH.sub.3)(CH.sub.2CH.sub.3),
and others. Thus the term `alkyl` includes primary alkyl groups,
secondary alkyl groups, and tertiary alkyl groups. Typical alkyl
groups include straight and branched chain alkyl groups having 1 to
12 carbon atoms, preferably 1-6 carbon atoms. The term `lower
alkyl` or "loweralkyl" and similar terms refer to alkyl groups
containing up to 6 carbon atoms.
[0034] The term "alkenyl" refers to alkyl groups as defined above,
wherein there is at least one carbon-carbon double bond, i.e.,
wherein two adjacent carbon atoms are attached by a double bond.
The term "alkynyl" refers to alkyl groups wherein two adjacent
carbon atoms are attached by a triple bond. Typical alkenyl and
alkynyl groups contain 2-12 carbon atoms, preferably 2-6 carbon
atoms. Lower alkenyl or lower alkynyl refers to groups having up to
6 carbon atoms. An alkenyl or alkynyl group may contain more than
one unsaturated bond, and may include both double and triple bonds,
but of course their bonding is consistent with well-known valence
limitations.
[0035] The term `alkoxy" refers to --OR, wherein R is alkyl.
[0036] As used herein, the term "halogen" or "halo" refers to
chloro, bromo, fluoro and iodo groups. Typical halo substituents
are F and/or Cl. "Haloalkyl" refers to an alkyl radical substituted
with one or more halogen atoms. The term "haloalkyl" thus includes
monohalo alkyl, dihalo alkyl, trihalo alkyl, perhaloalkyl, and the
like.
[0037] "Amino" refers herein to the group --NH.sub.2. The term
"alkylamino" refers herein to the group --NRR' where R and R' are
each independently selected from hydrogen or a lower alkyl,
provided --NRR' is not --NH.sub.2. The term "arylamino" refers
herein to the group --NRR' where R is aryl and R' is hydrogen, a
lower alkyl, or an aryl. The term "aralkylamino" refers herein to
the group --NRR' where R is a lower aralkyl and R' is hydrogen, a
loweralkyl, an aryl, or a loweraralkyl. The term cyano refers to
the group --CN. The term nitro refers to the group --NO.sub.2.
[0038] The term "alkoxyalkyl" refers to the group
-alk.sub.1-O-alk.sub.2 where alk.sub.1 is an alkyl or alkenyl
linking group, and alk.sub.2 is alkyl or alkenyl. The term
"loweralkoxyalkyl" refers to an alkoxyalkyl where alk.sub.1 is
loweralkyl or loweralkenyl, and alk.sub.2 is loweralkyl or
loweralkenyl. The term "aryl-oxyalkyl" refers to the group
-alkyl-O-aryl, where -alkyl- is a C.sub.1-12 straight or branched
chain alkyl linking group, preferably C.sub.1-6. The term
"aralkoxyalkyl" refers to the group -alkylenyl-O-aralkyl, where
aralkyl is preferably a loweraralkyl.
[0039] The term "aminocarbonyl" refers herein to the group
--C(O)--NH.sub.2. "Substituted aminocarbonyl" refers herein to the
group --C(O)--NRR' where R is loweralkyl and R' is hydrogen or a
loweralkyl. In some embodiments, R and R', together with the N atom
attached to them may be taken together to form a
"heterocycloalkylcarbonyl" group. The term "arylaminocarbonyl"
refers herein to the group --C(O)--NRR' where R is an aryl and R'
is hydrogen, loweralkyl or aryl. "aralkylaminocarbonyl" refers
herein to the group --C(O)--NRR' where R is loweraralkyl and R' is
hydrogen, loweralkyl, aryl, or loweraralkyl.
[0040] "Aminosulfonyl" refers herein to the group
--S(O).sub.2--NH.sub.2. "Substituted aminosulfonyl" refers herein
to the group --S(O).sub.2--NRR' where R is loweralkyl and R' is
hydrogen or a loweralkyl. The term "aralkylaminosulfonlyaryl"
refers herein to the group -aryl-S(O).sub.2--NH-- aralkyl, where
the aralkyl is loweraralkyl.
[0041] "Carbonyl" refers to the divalent group --C(O)--. "Carboxy"
refers to --C(.dbd.O)--OH. "Alkoxycarbonyl" refers to ester
--C(.dbd.O)--OR wherein R is optionally substituted lower alkyl.
"Loweralkoxycarbonyl" refers to ester --C(.dbd.O)--OR wherein R is
optionally substituted lower loweralkyl. "Cycloalkyloxycarbonyl"
refers to --C(.dbd.O)--OR wherein R is optionally substituted C3-C8
cycloalkyl.
[0042] "Cycloalkyl" refers to a mono- or polycyclic, carbocyclic
alkyl substituent. Carbocycloalkyl groups are cycloalkyl groups in
which all ring atoms are carbon. Typical cycloalkyl substituents
have from 3 to 8 backbone (i.e., ring) atoms. When used in
connection with cycloalkyl substituents, the term "polycyclic"
refers herein to fused and non-fused alkyl cyclic structures. The
term "partially unsaturated cycloalkyl", "partially saturated
cycloalkyl", and "cycloalkenyl" all refer to a cycloalkyl group
wherein there is at least one point of unsaturation, i.e., wherein
to adjacent ring atoms are connected by a double bond or a triple
bond. Such rings typically contain 1-2 double bonds for 5-6
membered rings, and 1-2 double bonds or one triple bond for 7-8
membered rings. Illustrative examples include cyclohexenyl,
cyclooctynyl, cyclopropenyl, cyclobutenyl, cyclohexadienyl, and the
like.
[0043] The term "heterocycloalkyl" refers herein to cycloalkyl
substituents that have from 1 to 5, and more typically from 1 to 4
heteroatoms as ring members in place of carbon atoms. Preferably,
heterocycloalkyl or "heterocyclyl" groups contain one or two
heteroatoms as ring members, typically only one heteroatom for 3-5
membered rings and 1-2 heteroatoms for 6-8 membered rings. Suitable
heteroatoms employed in heterocyclic groups of the present
invention are nitrogen, oxygen, and sulfur. Representative
heterocycloalkyl moieties include, for example, pyrrolidinyl,
tetrahydrofuranyl, oxirane, oxetane, oxepane, thiirane, thietane,
azetidine, morpholino, piperazinyl, piperidinyl and the like.
[0044] The terms "substituted heterocycle", "heterocyclic group" or
"heterocycle" as used herein refers to any 3- or 4-membered ring
containing a heteroatom selected from nitrogen, oxygen, and sulfur
or a 5- or 6-membered ring containing from one to three
heteroatoms, preferably 1-2 heteroatoms, selected from the group
consisting of nitrogen, oxygen, or sulfur; wherein the 5-membered
ring has 0-2 double bonds and the 6-membered ring has 0-3 double
bonds; wherein the nitrogen and sulfur atom maybe optionally
oxidized; wherein the nitrogen and sulfur heteroatoms may be
optionally quarternized; and including any bicyclic group in which
any of the above heterocyclic rings is fused to a benzene ring or
another 5- or 6-membered heterocyclic ring as described herein.
Preferred heterocycles include, for example: diazapinyl,
pyrrolinyl, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl,
imidazolidinyl, piperidinyl, piperazinyl, N-methyl piperazinyl,
azetidinyl, N-methylazetidinyl, oxazolidinyl, isoazolidinyl,
morpholinyl, thiazolidinyl, isothiazolidinyl, and oxiranyl. The
heterocyclic groups may be attached at various positions as will be
apparent to those having skill in the organic and medicinal
chemistry arts in conjunction with the disclosure herein.
[0045] The term `syn to one another` as used herein means the
groups being described are all on the same face of the ring they
are attached to, e.g., the groups are either all above the plane of
the ring, or they are all below the plane of the ring. Compounds of
Formula I can thus be depicted as (Ia) or (Ib) when Q is C:
##STR00003##
[0046] Heterocyclic moieties can be unsubstituted or they can be
substituted with one or more substituents independently selected
from hydroxy, halo, oxo (C.dbd.O), alkylimino (RN.dbd., wherein R
is a loweralkyl or loweralkoxy group), amino, alkylamino,
dialkylamino, acylaminoalkyl, alkoxy, thioalkoxy, lower
alkoxyalkoxy, loweralkyl, cycloalkyl or haloalkyl. Typically,
substituted heterocyclic groups will have up to four substituent
groups. The term "cyclic ether" as used herein refers to a 3-7
membered ring containing one oxygen atom (0) as a ring member.
Where the cyclic ether is "optionally substituted" it can be
substituted at any carbon atom with a group suitable as a
substituent for a heterocyclic group, typically up to three
substituents selected from lower alkyl, lower alkoxy, halo,
hydroxy, --C(O)-lower alkyl, and --C(O)-lower alkoxy. In preferred
embodiments, halo, hydroxy and lower alkoxy are not attached to the
carbon atoms of the ring that are bonded directly to the oxygen
atom in the cyclic ether ring. Specific examples include oxirane,
oxetane (e.g., 3-oxetane), tetrahydrofuran (including
2-tetrahydrofuranyl and 3-tetrahydrofuranyl), tetrahydropyran
(e.g., 4-tetrahydropyranyl), and oxepane.
[0047] "Aryl" refers to monocyclic and polycyclic aromatic groups
having from 5 to 14 backbone carbon or hetero atoms, and includes
both carbocyclic aryl groups and heteroaromatic aryl groups.
Carbocyclic aryl groups are aryl groups in which all ring atoms in
the aromatic ring are carbon, typically including phenyl and
naphthyl. Exemplary aryl moieties employed as substituents in
compounds of the present invention include phenyl, pyridyl,
pyrimidinyl, thiazolyl, indolyl, imidazolyl, oxadiazolyl,
tetrazolyl, pyrazinyl, triazolyl, thiophenyl, furanyl, quinolinyl,
purinyl, naphthyl, benzothiazolyl, benzopyridyl, and
benzimidazolyl, and the like. When used in connection with aryl
substituents, the term "polycyclic aryl" refers herein to fused and
non-fused cyclic structures in which at least one cyclic structure
is aromatic, such as, for example, benzodioxozolo (which has a
heterocyclic structure fused to a phenyl group, naphthyl, and the
like. Where "aryl" is used, the group is preferably a carbocyclic
group; the term "heteroaryl" is used for aryl groups when ones
containing one or more heteroatoms are preferred.
[0048] The term "heteroaryl" refers herein to aryl groups having
from 1 to 4 heteroatoms as ring atoms in an aromatic ring with the
remainder of the ring atoms being carbon atoms, in a 5-14 atom
aromatic ring system that can be monocyclic or polycyclic.
Monocyclic heteroaryl rings are typically 5-6 atoms in size.
Exemplary heteroaryl moieties employed as substituents in compounds
of the present invention include pyridyl, pyrimidinyl, thiazolyl,
indolyl, imidazolyl, oxadiazolyl, tetrazolyl, pyrazinyl, triazolyl,
thiophenyl, furanyl, quinolinyl, purinyl, benzothiazolyl,
benzopyridyl, and benzimidazolyl, and the like.
[0049] "Aralkyl" or "arylalkyl" refers to an aryl group connected
to a structure through an alkylene linking group, e.g., a structure
such as --(CH.sub.2).sub.1-4-Ar, where Ar represents an aryl group.
"Lower aralkyl" or similar terms indicate that the alkyl linking
group has up to 6 carbon atoms.
[0050] "Optionally substituted" or "substituted" refers to the
replacement of one or more hydrogen atoms with a monovalent or
divalent radical. Alkyl, alkenyl, alkynyl, cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl groups described herein may
be substituted or unsubstituted. Suitable substitution groups
include, for example, hydroxy, nitro, amino, imino, cyano, halo,
thio, sulfonyl, thioamido, amidino, imidino, oxo, oxamidino,
methoxamidino, imidino, guanidino, sulfonamido, carboxyl, formyl,
loweralkyl, haloloweralkyl, loweralkylamino, haloloweralkylamino,
loweralkoxy, haloloweralkoxy, loweralkoxyalkyl, alkylcarbonyl,
aminocarbonyl, arylcarbonyl, aralkylcarbonyl, heteroarylcarbonyl,
heteroaralkyl-carbonyl, alkylthio, aminoalkyl, cyanoalkyl, aryl and
the like, provided that oxo, imidino or other divalent substitution
groups are not placed on aryl or heteroaryl rings due to the well
known valence limitations of such rings.
[0051] The substitution group can itself be substituted where
valence permits, i.e., where the substitution group contains at
least one CH, NH or OH having a hydrogen atom that can be replaced.
The group substituted onto the substitution group can be carboxyl,
halo (on carbon only); nitro, amino, cyano, hydroxy, loweralkyl,
loweralkoxy, C(O)R, --OC(O)R, --OC(O)OR, --NRCOR, --CONR.sub.2,
--NRCOOR, --C(S)NR.sub.2, --NRC(S)R, --OC(O)NR.sub.2, --SR,
--SO.sub.3H, --SO.sub.2R or C3-8 cycloalkyl or 3-8 membered
heterocycloalkyl, where each R is independently selected from
hydrogen, lower haloalkyl, lower alkoxyalkyl, and loweralkyl, and
where two R on the same atom or on directly connected atoms can be
linked together to form a 5-6 membered heterocyclic ring.
[0052] When a substituted substituent includes a straight chain
group, the substitution can occur either within the chain (e.g.,
2-hydroxypropyl, 2-aminobutyl, and the like) or at the chain
terminus (e.g., 2-hydroxyethyl, 3-cyanopropyl, and the like).
Substituted substituents can be straight chain, branched or cyclic
arrangements of covalently bonded carbon or heteroatoms.
[0053] It is understood that the above definitions are not intended
to include impermissible substitution patterns (e.g., methyl
substituted with five fluoro groups or a halogen atom substituted
with another halogen atom). Such impermissible substitution
patterns are well known to the skilled artisan.
[0054] "Syn" as used herein has its ordinary meaning, and is used
in connection with Formula I to indicate that the specified groups
are attached to sp.sup.a hybridized (tetrahedral) carbon centers
and extend out from one face of the cyclohexyl or piperidinyl ring,
i.e., those groups all project toward the `alpha` face of the ring,
or they all project toward the `beta` face of the ring. This is
thus used as a convenient way to define the relative orientations
of two or more groups, without limiting the compounds to a specific
chiral configuration. This reflects the fact that the compounds of
the invention have such groups in a specific relative orientation,
but are not limited to either enantiomer of that specific relative
orientation. Accordingly, unless described as optically active,
such compounds may be racemic, but also include each of the two
enantiomers having the specified relative stereochemistry. In some
embodiments, the compounds of the invention are optically active
form as further described herein, and in preferred embodiments of
the invention, the compounds are obtained and used in optically
active form. Preferably, the enantiomer having greater potency as
an inhbitor of at least two of Pim1, Pim2 and Pim3 is selected.
[0055] It will also be apparent to those skilled in the art that
the compounds of the invention, as well as the pharmaceutically
acceptable salts, esters, metabolites and prodrugs of any of them,
may be subject to tautomerization and may therefore exist in
various tautomeric forms wherein a proton of one atom of a molecule
shifts to another atom and the chemical bonds between the atoms of
the molecules are consequently rearranged. See, e.g., March,
Advanced Organic Chemistry: Reactions, Mechanisms and Structures,
Fourth Edition, John Wiley & Sons, pages 69-74 (1992). As used
herein, the term "tautomer" refers to the compounds produced by the
proton shift, and it should be understood that all tautomeric
forms, insofar as they may exist, are included within the
invention.
[0056] The compounds of the invention comprise one or more
asymmetrically substituted carbon atoms. Such asymmetrically
substituted carbon atoms can result in the compounds of the
invention existing in enantiomers, diastereomers, and other
stereoisomeric forms that may be defined, in terms of absolute
stereochemistry, such as in (R)- or (S)-forms. The compounds of the
invention are sometimes depicted herein as single enantiomers, and
are intended to encompass the specific configuration depicted and
the enantiomer of that specific configuration (the mirror image
isomer of the depicted configuration), unless otherwise specified.
The depicted structures herein describe the relative
stereochemistry of the compounds where two or more chiral centers,
but the invention is not limited to the depicted enantiomer's
absolute stereochemistry unless otherwise stated. The invention
includes both enantiomers, each of which will exhibit Pim
inhibition, even though one enantiomer will be more potent than the
other. In some instances, compounds of the invention have been
synthesized in racemic form and separated into individual isomers
by chiral chromatography or similar conventional methods, and the
analytical data about the two enantiomers do not provide definitive
information about absolute stereochemical configuration. In such
cases, the absolute stereochemistry of the most active enantiomer
has been identified based on correlation with similar compounds of
known absolute stereochemistry, rather than by a definitive
physical method such as X-ray crystallography. Therefore, in
certain embodiments, the preferred enantiomer of a compound
described herein is the specific isomer depicted or its opposite
enantiomer, whichever has the lower IC-50 for Pim kinase inhibition
using the assay methods described herein, i.e., the enantiomer that
is more potent as a Pim inhibitor for at least two of Pim1, Pim2,
and Pim3.
[0057] The terms "S" and "R" configuration, as used herein, are as
defined by the IUPAC 1974 RECOMMENDATIONS FOR SECTION E,
FUNDAMENTAL STEREOCHEMISTRY, Pure Appl. Chem. 45:13-30 (1976). The
terms .alpha. and .beta. are employed for ring positions of cyclic
compounds. The .alpha.-side of the reference plane is that side on
which the preferred substituent lies at the lower numbered
position. Those substituents lying on the opposite side of the
reference plane are assigned .beta. descriptor. It should be noted
that this usage differs from that for cyclic stereoparents, in
which ".alpha." means "below the plane" and denotes absolute
configuration. The terms .alpha. and .beta. configuration, as used
herein, are as defined by the CHEMICAL ABSTRACTS INDEX
GUIDE-APPENDIX IV (1987) paragraph 203.
[0058] As used herein, the term "pharmaceutically acceptable salts"
refers to the nontoxic acid or base addition salts of the compounds
of Formula I or II, wherein the compound acquires a positive or
negative charge as a result of adding or removing a proton; the
salt then includes a counterion of opposite charge from the
compound itself, and the counterion is preferably one suitable for
pharmaceutical administration under the conditions where the
compound would be used. These salts can be prepared in situ during
the final isolation and purification of the compounds of Formula I
or II, or by separately reacting the base or acid functions with a
suitable organic or inorganic acid or base, respectively.
Representative salts include but are not limited to the following:
acetate, adipate, alginate, citrate, aspartate, benzoate,
benzenesulfonate, bisulfate, butyrate, camphorate,
camphorsulfonate, digluconate, cyclopentanepropionate,
dodecylsulfate, ethanesulfonate, glucoheptanoate, glycerophosphate,
hemisulfate, heptanoate, hexanoate, fumarate, hydrochloride,
hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate,
maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate,
oxalate, pamoate, pectinate, persulfate, 3-phenylproionate,
picrate, pivalate, propionate, succinate, sulfate, tartrate,
thiocyanate, p-toluenesulfonate and undecanoate.
[0059] Also, a basic nitrogen-containing group in compounds of the
invention can be quaternized with such agents as loweralkyl
halides, such as methyl, ethyl, propyl, and butyl chloride,
bromides, and iodides; dialkyl sulfates like dimethyl, diethyl,
dibutyl, and diamyl sulfates, long chain halides such as decyl,
lauryl, myristyl and stearyl chlorides, bromides and iodides,
aralkyl halides like benzyl and phenethyl bromides, and others.
Water or oil-soluble or dispersible products are thereby obtained.
These quaternized ammonium salts when paired with a
pharmaceutically acceptable anion can also serve as
pharmaceutically acceptable salts.
[0060] Examples of acids which may be employed to form
pharmaceutically acceptable acid addition salts include such
inorganic acids as hydrochloric acid, sulfuric acid and phosphoric
acid and such organic acids as oxalic acid, maleic acid,
methanesulfonic acid, succinic acid and citric acid. Basic addition
salts can be prepared in situ during the final isolation and
purification of the compounds of formula (I), or separately by
reacting carboxylic acid moieties with a suitable base such as the
hydroxide, carbonate or bicarbonate of a pharmaceutically
acceptable metal cation or with ammonia, or an organic primary,
secondary or tertiary amine. Counterions for pharmaceutically
acceptable salts include, but are not limited to, cations based on
the alkali and alkaline earth metals, such as sodium, lithium,
potassium, calcium, magnesium, aluminum salts and the like, as well
as nontoxic ammonium, quaternary ammonium, and amine cations,
including, but not limited to ammonium, tetramethylammonium,
tetraethylammonium, methyl-amine, dimethylamine, trimethylamine,
triethylamine, ethylamine, and the like. Other representative
organic amines useful for the formation of base addition salts
include diethylamine, ethylenediamine, ethanolamine,
diethanolamine, piperazine and the like.
[0061] As used herein, the term "pharmaceutically acceptable ester"
refers to esters, which hydrolyze in vivo and include those that
break down readily in the human body to leave the parent compound
or a salt thereof. Suitable ester groups include, for example,
those derived from pharmaceutically acceptable aliphatic carboxylic
acids, particularly alkanoic, alkenoic, cycloalkanoic and
alkanedioic acids, in which each alkyl or alkenyl moiety
advantageously has not more than 6 carbon atoms. Examples of
particular pharmaceutically acceptable esters include formates,
acetates, propionates, maleates, lactates, hydroxyacetates,
butyrates, acrylates and ethylsuccinates.
[0062] The term "pharmaceutically acceptable prodrugs" as used
herein refers to those prodrugs of the compounds of the present
invention which are, within the scope of sound medical judgment,
suitable for use in contact with the tissues of humans and lower
animals without undue toxicity, irritation, allergic response, and
the like, commensurate with a reasonable benefit/risk ratio, and
effective for their intended use, as well as the zwitterionic
forms, where possible, of the compounds of the invention. The term
"prodrug" refers to compounds that are rapidly transformed in vivo
to yield the parent compound of the above formula, for example by
hydrolysis in blood. A thorough discussion is provided in T.
Higuchi and V. Stella, PRO-DRUGS AS NOVEL DELIVERY SYSTEMS, Vol. 14
of the A.C.S. Symposium Series, and in Edward B. Roche, ed.,
BIOREVERSIBLE CARRIERS IN DRUG DESIGN, American Pharmaceutical
Association and Pergamon Press, 1987, both of which are
incorporated herein by reference.
[0063] It will be apparent to those skilled in the art that the
compounds of the invention, or their tautomers, prodrugs and
stereoisomers, as well as the pharmaceutically acceptable salts,
esters and prodrugs of any of them, may be processed in vivo
through metabolism in a human or animal body or cell to produce
metabolites. The term "metabolite" as used herein refers to the
formula of any derivative produced in a subject after
administration of a parent compound. The derivatives may be
produced from the parent compound by various biochemical
transformations in the subject such as, for example, oxidation,
reduction, hydrolysis, or conjugation and include, for example,
oxides and demethylated derivatives. The metabolites of a compound
of the invention may be identified using routine techniques known
in the art. See, e.g., Bertolini, G. et al., J. Med. Chem.
40:2011-2016 (1997); Shan, D. et al., J. Pharm. Sci. 86(7):765-767;
Bagshawe K., Drug Dev. Res. 34:220-230 (1995); Bodor, N., Advances
in Drug Res. 13:224-331 (1984); Bundgaard, H., Design of Prodrugs
(Elsevier Press 1985); and Larsen, I. K., Design and Application of
Prodrugs, Drug Design and Development (Krogsgaard-Larsen et al.,
eds., Harwood Academic Publishers, 1991). It should be understood
that individual chemical compounds that are metabolites of the
compounds of formula (I) or their tautomers, prodrugs and
stereoisomers, as well as the pharmaceutically acceptable salts,
esters and prodrugs of any of them, are included within the
invention.
[0064] In one aspect, the invention provides compounds of Formula
A:
##STR00004##
[0065] wherein:
[0066] groups attached to the ring containing Q that are depicted
inside the ring are all syn to each other, and all groups attached
to that ring that are depicted outside the ring are syn to one
another;
[0067] Q is C or N;
[0068] R.sup.q is H when Q is C, and R.sup.q is absent when Q is
N;
[0069] R.sup.1 is selected from H, C.sub.1-4 alkyl, C.sub.3-6
cycloalkyl, C.sub.4-6 heterocyclyl, --(CR'.sub.2).sub.1-3--OR' and
--OR',
[0070] where each R' is independently H or C.sub.1-4 alkyl, and
each alkyl, cycloalkyl and heterocyclyl is optionally substituted
with up to two groups selected from halo, CN, NH.sub.2, hydroxy,
oxo, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy, and C.sub.1-4
haloalkoxy; R.sup.1b, R.sup.1c, and R.sup.1d are each independently
selected from H, halo, OR', --(CH.sub.2).sub.1-2OR', and
CONR'.sub.2;
[0071] one of R.sup.2a and R.sup.2b is H, and the other of R.sup.2a
and R.sup.2b is selected from CN, halo, azido, amino, --OR,
--O(CH.sub.2).sub.1-3OR, --NRC(O)R, --NRC(O)OR, --NHSO.sub.2R,
--SO.sub.2R, --OSO.sub.2R, --SR, --S(O)R, --OP(O)R.sub.2, and
1-pyridonyl or 1-triazolyl optionally substituted with up to two
groups selected from halo, C.sub.1-4 alkyl, hydroxy-substituted
C.sub.1-4 alkyl, C.sub.2-4 alkenyl, C.sub.3-6 cycloalkyl, C.sub.1-4
alkoxy, and C.sub.5-6 heteroaryl;
[0072] where each R is independently C.sub.5-6 heteroaryl or
C.sub.1-4 alkyl optionally substituted with up to three groups
selected from cyano, halo, hydroxy, carboxy, C.sub.1-4
alkylsulfonyl, and C.sub.1-4 alkoxy;
[0073] or R.sup.2a and R.sup.2b taken together may form a dialkyl
ketal or 5-6 membered cyclic ketal, .dbd.O or .dbd.N--OR'', where
R'' is H or C.sub.1-4 alkyl;
[0074] Ring A is selected from pyridinyl, pyrimidinyl, pyrazinyl,
and thiazolyl, each having N positioned as shown in Formula (I);
and
[0075] Ring A is optionally substituted with 1 or 2 groups selected
from halo, CN, NH.sub.2, hydroxy, C.sub.1-4 alkyl,
C.sub.1-4haloalkyl, C.sub.1-4alkoxy, and C.sub.1-4haloalkoxy;
or a pharmaceutically acceptable salt thereof
[0076] In some embodiments of the compounds of Formula (A),
R.sup.1c or R.sup.1d or both R.sup.1c and R.sup.1d represent halo,
preferably F. Typically, one of R.sup.1 and R.sup.1b, but not both,
represents H. In some embodiments, R.sup.1 is H and R.sup.1b is
halo or CONR'.sub.2, such as --CONHR' where R' is C.sub.1-4 alkyl.
In other embodiments, R.sup.1b is H, in which case R.sup.1 is
preferably selected from C.sub.1-4 alkyl, C.sub.3-6 cycloalkyl,
C.sub.4-6 heterocyclyl, --(CR'.sub.2).sub.1-3--OR' and --OR', where
each R' is independently H or C.sub.1-4 alkyl, and each alkyl,
cycloalkyl and heterocyclyl is optionally substituted with up to
two groups selected from halo, CN, NH.sub.2, hydroxy, and
C.sub.1-4alkoxy.
[0077] In preferred embodiments of the compounds of Formula (A), Q
is CH. In many embodiments of these compounds, Ring A is a pyridine
or a thiazole ring. Typically, these compounds are of this
formula:
##STR00005##
where Z.sup.3 is H or NH.sub.2, or of this formula
##STR00006##
[0078] wherein Z.sup.3 is H or NH.sub.2,
[0079] and R.sup.1, R.sup.1b, R.sup.1C, R.sup.1d, R.sup.2a and
R.sup.2b are as defined above for any of the embodiments of
compounds of Formula (A).
[0080] Specific embodiments of these compounds include Formula A2
and A3:
##STR00007##
where each Z.sup.2 is H or amino, Y is halo or H, and R.sup.1,
R.sup.1b, R.sup.1c and R.sup.2a are as defined above for any of the
embodiments of compounds of Formula (A). In some embodiments, for
example, R.sup.1c is F, and R.sup.1b is H.
[0081] In some of these embodiments, R.sup.2a is preferably
methoxy. In other embodiments wherein R.sup.2b is H, R.sup.2a is
--OMe or --O(CH.sub.2).sub.2--X, wherein X is --OMe, COOH, CN or
--SO.sub.2Me, or R.sup.2a is 1-triazolyl (e.g., 1,2,3-triazolyl)
that is optionally substituted with up to two groups selected from
halo, C.sub.1-4 alkyl, hydroxy-substituted C.sub.1-4 alkyl,
C.sub.2-4 alkenyl, and C.sub.1-4 alkoxy, or --SO.sub.2Me.
[0082] In some embodiments of these compounds of Formula (A), A1,
A2, A3 or A4, R.sup.2a is --OMe or --O(CH.sub.2).sub.2--X, wherein
X is --OMe, COOH, CN or --SO.sub.2Me; preferably R.sup.2a is --OMe
or --OCH.sub.2CH.sub.2CN. R.sup.1 in some embodiments of these
compounds can be selected from 2-hydroxy-2-propyl, methoxymethyl,
ethoxymethyl, methoxy, ethoxy, isopropoxy, cyclopropyl, cyclobutyl,
1-hydroxycyclobutyl, cyclopentyl, tetrahydropyranyl, 4-f,
4-hydroxy-4-tetrahydropyranyl, 4-tetrahydropyranyloxy, and
4-tetrahydropyranyl.
[0083] In certain embodiments, the compounds of Formula (A) have
the Formula (I):
##STR00008##
[0084] R.sup.1 is selected from H, C.sub.1-4 alkyl, C.sub.3-6
cycloalkyl, C.sub.4-6 heterocyclyl, --(CR'.sub.2).sub.1-3--OR' and
--OR',
[0085] where each R' is independently H or C.sub.1-4 alkyl,
[0086] and each alkyl, cycloalkyl and heterocyclyl is optionally
substituted with up to two groups selected from halo, CN, NH.sub.2,
hydroxy, oxo, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy, and C.sub.1-4
haloalkoxy;
[0087] one of R.sup.2a and R.sup.2b is H,
[0088] and the other of R.sup.2a and R.sup.2b is selected from CN,
halo, azido, amino, --OR, --O(CH.sub.2).sub.1-3 OR, --NRC(O)R,
--NRC(O)OR, --NHSO.sub.2R, --SO.sub.2R, --OSO.sub.2R, --SR,
--S(O)R, --OP(O)R.sub.2, and triazolyl optionally substituted with
up to two groups selected from halo, C.sub.1-4 alkyl,
hydroxy-substituted C.sub.1-4 alkyl, C.sub.3-6 cycloalkyl,
C.sub.1-4 alkoxy, and C.sub.5-6 heteroaryl;
[0089] where each R is independently C.sub.1-4 alkyl optionally
substituted with up to three groups selected from cyano, halo,
hydroxy, carboxy, C.sub.1-4 alkylsulfonyl, and C.sub.1-4
alkoxy;
[0090] or R.sup.2a and R.sup.2b taken together form a dialkyl ketal
or 5-6 membered cyclic ketal, .dbd.O or .dbd.N--OR'', where R'' is
H or C.sub.1-4 alkyl;
[0091] ring A is selected from pyridinyl, pyrimidinyl, pyrazinyl,
and thiazolyl, each having N positioned as shown in Formula (I);
and
[0092] Ring A is optionally substituted with 1 or 2 groups selected
from halo, CN, NH.sub.2, hydroxy, C.sub.1-4 alkyl,
C.sub.1-4haloalkyl, C.sub.1-4alkoxy, and C.sub.1-4 haloalkoxy;
[0093] or a pharmaceutically acceptable salt thereof.
[0094] The following enumerated embodiments represent additional
aspects and variations of the invention:
1. A compound of Formula (A)
##STR00009##
[0095] wherein:
[0096] groups attached to the ring containing Q that are depicted
inside the ring are all syn to each other, and all groups attached
to that ring that are depicted outside the ring are syn to one
another;
[0097] Q is C or N;
[0098] R.sup.q is H when Q is C, and R.sup.q is absent when Q is
N;
[0099] R.sup.1 is selected from H, C.sub.1-4 alkyl, C.sub.3-6
cycloalkyl, C.sub.4-6 heterocyclyl, --(CR'.sub.2).sub.1-3--OR' and
--OR',
[0100] where each R' is independently H or C.sub.1-4 alkyl,
[0101] and each alkyl, cycloalkyl and heterocyclyl is optionally
substituted with up to two groups selected from halo, CN, NH.sub.2,
hydroxy, oxo, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy, and C.sub.1-4
haloalkoxy;
[0102] R.sup.1b, R.sup.1c, and R.sup.1d are each independently
selected from H, halo, OR', R', --(CH.sub.2).sub.1-2OR', and
CONR'.sub.2;
[0103] one of R.sup.2a and R.sup.2b is H,
[0104] and the other of R.sup.2a and R.sup.2b is selected from CN,
halo, azido, amino, --OR, --O(CH.sub.2).sub.1-3 OR, --NRC(O)R,
--NRC(O)OR, --NHSO.sub.2R, --SO.sub.2R, --OSO.sub.2R, --SR,
--S(O)R, --OP(O)R.sub.2, and 1-pyridonyl or 1-triazolyl optionally
substituted with up to two groups selected from halo, C.sub.1-4
alkyl, hydroxy-substituted C.sub.1-4 alkyl, C.sub.2-4 alkenyl,
C.sub.3-6 cycloalkyl, C.sub.1-4 alkoxy, and C.sub.5-6
heteroaryl;
[0105] where each R is independently C.sub.5-6 heteroaryl or
C.sub.1-4 alkyl optionally substituted with up to three groups
selected from cyano, halo, hydroxy, carboxy, C.sub.1-4
alkylsulfonyl, and C.sub.1-4 alkoxy;
[0106] or R.sup.2a and R.sup.2b taken together may form a dialkyl
ketal or 5-6 membered cyclic ketal, .dbd.O or .dbd.N--OR'', where
R'' is H or C.sub.1-4 alkyl;
[0107] ring A is selected from pyridinyl, pyrimidinyl, pyrazinyl,
and thiazolyl, each having N positioned as shown in Formula (I);
and
[0108] Ring A is optionally substituted with 1 or 2 groups selected
from halo, CN, NH.sub.2, hydroxy, C.sub.1-4 alkyl, C.sub.1-4
haloalkyl, C.sub.1-4 alkoxy, and C.sub.1-4 haloalkoxy; [0109] or a
pharmaceutically acceptable salt thereof 2. The compound of Formula
(A) according to embodiment 1, wherein one but not both, of
R.sup.1b and R.sup.1 represents H. 3. The compound of embodiment 1,
wherein R.sup.1d is F. 4. The compound of embodiment 1 or 2,
wherein R.sup.1c is F. 5. The compound of any of embodiments 1-3,
wherein R.sup.1b is H. 6. The compound of any of embodiments 1-3,
wherein R.sup.1b is H or CONR'.sub.2. 7. The compound of any of
embodiments 1-6, wherein R.sup.2b is H. 8. The compound of any of
embodiments 1-7, wherein R.sup.2a is --OMe, --SO.sub.2Me,
--NHCOOMe, or --O(CH.sub.2).sub.2--X, wherein X is --OMe, COOH, CN
or --SO.sub.2Me, or R.sup.2a is 1-triazolyl (e.g., 1,2,3-triazolyl)
or 1-pyridonyl that is optionally substituted with up to two groups
selected from halo, C.sub.1-4 alkyl, hydroxy-substituted C.sub.1-4
alkyl, C.sub.2-4 alkenyl, C.sub.3-6 cycloalkyl, and C.sub.1-4
alkoxy, or --SO.sub.2Me. 9. The compound of any of embodiments 1-8,
wherein R.sup.1 is selected from 2-hydroxy-2-propyl, methoxymethyl,
ethoxymethyl, methoxy, ethoxy, isopropoxy, cyclopropyl, cyclobutyl,
1-hydroxycyclobutyl, cyclopentyl, tetrahydropyranyl, 4-F,
4-hydroxy-4-tetrahydropyranyl, 4-tetrahydropyranyloxy, and
4-tetrahydropyranyl. 10. The compound of any of embodiments 1-6,
wherein R.sup.2b is OMe. 11. The compound of embodiment 1, which is
a compound of Formula (I):
##STR00010##
[0110] wherein:
[0111] groups attached to the ring containing Q that are depicted
inside the ring are all syn to each other, and all groups attached
to that ring that are depicted outside the ring are syn to one
another;
[0112] Q is C or N;
[0113] R.sup.q is H when Q is C, and R.sup.q is absent when Q is
N;
[0114] R.sup.1 is selected from H, C.sub.1-4 alkyl, C.sub.3-6
cycloalkyl, C.sub.4-6 heterocyclyl, --(CR'.sub.2).sub.1-3--OR' and
--OR',
[0115] where each R' is independently H or C.sub.1-4 alkyl,
[0116] and each alkyl, cycloalkyl and heterocyclyl is optionally
substituted with up to two groups selected from halo, CN, NH.sub.2,
hydroxy, oxo, C.sub.1-4 haloalkyl, C.sub.1-4alkoxy, and C.sub.1-4
haloalkoxy;
[0117] one of R.sup.2a and R.sup.2b is H,
[0118] and the other of R.sup.2a and R.sup.2b is selected from CN,
halo, azido, amino, --OR, --O(CH.sub.2).sub.1-3 OR, --NRC(O)R,
--NRC(O)OR, --NHSO.sub.2R, --SO.sub.2R, --OSO.sub.2R, --SR,
--S(O)R, --OP(O)R.sub.2, and N-pyridonyl or 1-triazolyl optionally
substituted with up to two groups selected from halo, C.sub.1-4
alkyl, hydroxy-substituted C.sub.1-4 alkyl, C.sub.2-4 alkenyl,
C.sub.3-6 cycloalkyl, C.sub.1-4 alkoxy, and C.sub.5-6
heteroaryl;
[0119] where each R is independently C.sub.5-6 heteroaryl or
C.sub.1-4 alkyl optionally substituted with up to three groups
selected from cyano, halo, hydroxy, carboxy, C.sub.1-4
alkylsulfonyl, and C.sub.1-4 alkoxy;
[0120] or R.sup.2a and R.sup.2b taken together may form a dialkyl
ketal or 5-6 membered cyclic ketal, .dbd.O or .dbd.N--OR'', where
R'' is H or C.sub.1-4 alkyl;
[0121] Ring A is selected from pyridinyl, pyrimidinyl, pyrazinyl,
and thiazolyl, each having N positioned as shown in Formula (I);
and
[0122] Ring A is optionally substituted with 1 or 2 groups selected
from halo, CN, NH.sub.2, hydroxy, C.sub.1-4 alkyl, C.sub.1-4
haloalkyl, C.sub.1-4 alkoxy, and C.sub.1-4 haloalkoxy;
[0123] or a pharmaceutically acceptable salt thereof.
12. The compound of embodiment 11, wherein R.sup.2a is H. 13. The
compound of embodiment 11, wherein R.sup.2b is H. 14. The compound
of embodiment 11 or 13, wherein R.sup.2a is --NHCOOMe. 15. The
compound of embodiment 11 or 13, wherein R.sup.2a is
--O(CH.sub.2).sub.2--CN. 16. The compound of embodiment 11 or 13,
wherein R.sup.2a is --O(CH.sub.2).sub.2--SO.sub.2Me. 17. The
compound of embodiment 11 or 13, wherein R.sup.2a is --OMe. 18. The
compound of embodiment 11 or 13, wherein R.sup.2a is --SO.sub.2Me.
19. The compound of and embodiment 11 or 12, wherein R.sup.2b is
--OMe. 20. The compound of embodiment 11, wherein R.sup.2a and
R.sup.2b taken together form.dbd.O or .dbd.N--OR'', where R'' is H
or C.sub.1-4 alkyl. 21. The compound of any of embodiments 11-20,
wherein R.sup.1 is selected from H, methyl, ethyl, isopropyl,
2-hydroxy-2-propyl, methoxymethyl, ethoxymethyl, methoxy, ethoxy,
isopropoxy, cyclopropyl, cyclobutyl, 1-hydroxycyclobutyl,
cyclopentyl, tetrahydrofuranyl, 4-fluoro-4-tetrahydrofuranyl,
tetrahydrothiopyran, and 4-tetrahydrothiopyran-1,1-dioxide. 22. The
compound of embodiment 21, wherein R.sup.1 is selected from H, Me,
tetrahydropyran, methoxymethyl, and ethoxymethyl. 23. The compound
of any one of the preceding embodiments, wherein Ring A is
pyridine, and is optionally substituted with F or amino. 24. The
compound of any one of embodiments 11-21, wherein Ring A is a
thiazolyl, optionally substituted with amino. 25. The compound of
any of embodiments 11-24, wherein R.sup.2a is selected from the
group consisting of --OR, --OCH.sub.2CH.sub.2OR,
--OCH.sub.2CH.sub.2CN, --OCH.sub.2CH.sub.2COOH,
--OCH.sub.2CH.sub.2SO.sub.2R, --CN, --NHC(O)OR, --NHC(O)R, N.sub.3,
NH.sub.2, F, --NHSO.sub.2R, --SO.sub.2R, --SR, --S(O)R,
unsubstituted triazole, and triazole substituted with Me, ethyl,
cyclopropyl, hydroxymethyl, C.sub.2-4 alkenyl, or thienyl; wherein
each R is methyl, ethyl or isopropyl. 26. The compound of any one
of embodiments 11-24, wherein R.sup.2b is selected from the group
consisting of --OR, --OCH.sub.2CH.sub.2OR, --OCH.sub.2CH.sub.2CN,
--OCH.sub.2CH.sub.2COOH, --OCH.sub.2CH.sub.2SO.sub.2R, --CN,
--NHC(O)OR, --NHC(O)R, N.sub.3, NH.sub.2, F, --NHSO.sub.2R,
--SO.sub.2R, --SR, --S(O)R, unsubstituted triazole, and triazole
substituted with Me, ethyl, cyclopropyl, hydroxymethyl, C.sub.2-4
alkenyl, or thienyl;
[0124] wherein each R is methyl, ethyl or isopropyl.
27. The compound of embodiment 11, wherein R.sup.2a is
--SO.sub.2Me, --OCH.sub.2CH.sub.2CN, 1,2,4-triazol-1-yl,
--OCH.sub.2CH.sub.2COOH, --OCH.sub.2CH.sub.2SO.sub.2R, --CN, or
--NHC(O)OMe. 28. The compound of embodiment 12, wherein R.sup.2b is
--OMe, --CN, --OCH.sub.2CH.sub.2CN, 1,2,4-triazol-1-yl,
--OCH.sub.2CH.sub.2COOH, --OCH.sub.2CH.sub.2SO.sub.2R, --CN, or
--NHC(O)OMe. 29. The compound of any of the preceding embodiments,
wherein Q is C. 30. The compound of any of embodiments 1-28,
wherein Q is N. 31. The compound of any of the preceding
embodiments, which is optically active and has a lower IC-50 than
its opposite enantiomer on at least one Pim kinase. 32. The
compound of embodiment 31, wherein the at least one Pim kinase is
Pim2 kinase. 33. The compound of any one of embodiments 11-32,
which is an optically active compound of Formula IIa or IIb:
[0125] or
##STR00011##
wherein Y is H or F, and Z is H or NH.sub.2. 34. The compound of
any one of embodiments 11-32, which is an optically active compound
of Formula IIIa or IIIb:
##STR00012##
wherein Y is H or F, and Z is H or NH.sub.2. 35. The compound of
embodiment 33, which is a compound of Formula IIa. 36. The compound
of embodiment 33, which is a compound of Formula IIb. 37. The
compound of embodiment 34, which is a compound of Formula IIa. 38.
The compound of embodiment 34, which is a compound of Formula IIb.
39. A compound selected from the group consisting of the compounds
in Table 1 and Table 2, and the pharmaceutically acceptable salts
thereof 40. A pharmaceutical composition comprising a compound of
any of embodiments 1-39, admixed with at least one pharmaceutically
acceptable excipient. 41. The pharmaceutical composition of
embodiment 40 which further comprises an additional agent for
treatment of cancer. 42. The pharmaceutical composition of
embodiment 41, wherein the additional therapeutic agent is selected
from MEK inhibitors, irinotecan, topotecan, gemcitabine,
5-fluorouracil, cytarabine, daunorubicin, PI3 Kinase inhibitors,
mTOR inhibitors, DNA synthesis inhibitors, leucovorin, carboplatin,
cisplatin, taxanes, tezacitabine, cyclophosphamide, vinca
alkaloids, imatinib, anthracyclines, rituximab, lenalidomide,
bortezomib and trastuzumab 43. A compound of any of embodiments
1-39 for use in the treatment of a condition that responds to
inhibitors of Provirus Integration of Moloney Kinase (PIM Kinase)
activity. 44. The compound according to embodiment 43, wherein the
condition is a cancer. 45. The compound according to embodiment 44,
wherein the cancer is selected from carcinoma of the lungs,
pancreas, thyroid, ovaries, bladder, breast, prostate or colon,
melanoma, myeloid leukemia, multiple myeloma, erythro leukemia,
villous colon adenoma, and osteosarcoma. 46. The compound of
embodiment 45, wherein the condition is an autoimmune disorder. 47.
A method of treating a disease or condition mediated by PIM kinase,
comprising administering to a subject in need thereof a
therapeutically effective amount of a compound according to any one
of embodiments 1-39, or a pharmaceutically acceptable salt thereof.
48. The method of embodiment 47, wherein the disease is selected
from carcinoma of the lungs, pancreas, thyroid, ovaries, bladder,
breast, prostate or colon, melanoma, myeloid leukemia, multiple
myeloma, erythro leukemia, villous colon adenoma, and osteosarcoma;
or the disease is an autoimmune disorder. 49. The method of
embodiment 48, wherein the disease is an autoimmune disorder. 50.
The method of embodiment 49, wherein the autoimmune disorder is
selected from Crohn's disease, inflammatory bowel disease,
rheumatoid arthritis, and chronic inflammatory diseases.
[0126] In some embodiments, at least one substituent for Ar is
selected from F, Cl, NH.sub.2, Me, Et, OMe, OEt, OCF.sub.3,
OCHF.sub.2, OCH.sub.2CF.sub.3, CN, CF.sub.3, SMe, SOMe, SO.sub.2Me,
--COOMe, --C(O)Me, --C(Me).sub.2-OH, MeOCH.sub.2--, HOCH.sub.2--,
hydroxyethyl, hydroxyethoxy, methoxyethyl, methoxyethoxy, oxetanyl
(e.g., 3-oxetanyl), isopropoxy, tetrahydropyranyloxy (e.g.,
4-tetrahydropyranyloxy), cyclopropyl, and CN. At least one
substituent for Ar is preferably selected from Me, F, NH.sub.2,
OMe, MeOCH.sub.2--, HOCH.sub.2--, hydroxyethyl, hydroxyethoxy,
methoxyethyl, methoxyethoxy, and CN.
[0127] These compounds may be used in racemic form, or the
individual enantiomers may be used, or mixtures of the enantiomers
may be used. Each enantiomer can be used, and preferably the
compound to be used is the enantiomer that has greater activity as
a Pim inhibitor.
[0128] The cyclohexyl or piperidine ring in these compounds has
three non-hydrogen substituents, not counting its attachment to the
pyridinyl ring in Formula I. The invention provides novel
combinations of substituents and their relative stereochemical
orientation on the cyclohexyl or piperidineyl ring, to provide
advantageous biological activities. Advantages provided by
preferred compounds include reduced drug-drug interactions, due to
reduction of time-dependent Cyp inhibition or pharmacokinetic
superiority based on improved clearance and metabolic
properties.
[0129] In one preferred embodiment of the claimed invention, the
compound is of formula IIc or IId,
##STR00013##
[0130] wherein Y is H or F, and Z.sup.2 is H or NH.sub.2; or. In
these compounds, R.sup.2a is preferably --OR,
--O(CH.sub.2).sub.1-3OR, --SO.sub.2R, or --NRC(O)OR, where each R
is independently C.sub.1-4 alkyl optionally substituted with cyano,
halo, hydroxy, carboxy, C.sub.1-4 alkylsulfonyl, or C.sub.1-4
alkoxy; and R.sup.2b is H. In some such embodiments, R.sup.2a is
preferably --OR, --O(CH.sub.2).sub.1-3OR, or --NRC(O)OR, where each
R is independently C.sub.1-4 alkyl optionally substituted with
cyano, halo, hydroxy, carboxy, C.sub.1-4 alkylsulfonyl, or
C.sub.1-4 alkoxy; and R.sup.2b is H In some preferred embodiments,
R.sup.2a is OMe. In these compounds, R.sup.1 can be H, Me,
4-tetrahydropyranyl, or 2-hydroxy-2-propyl. In specific
embodiments, R.sup.2a is --OMe, --OCH.sub.2CH.sub.2CN,
--OCH.sub.2CH.sub.2OMe, or --OCH.sub.2CH.sub.2SO.sub.2Me or
--NHCOOMe, or --N(Me)COOMe. Preferably when R.sup.2b is H, R.sup.1
is not H.
[0131] In another embodiment, the compound is of Formula IIe or
IIf:
##STR00014##
[0132] wherein Y is H or F, and Z.sup.2 is H or NH.sub.2. In these
compounds, R.sup.2a is preferably --OR, --O(CH.sub.2).sub.1-3OR,
--SO.sub.2R, or --NRC(O)OR, where each R is independently C.sub.1-4
alkyl optionally substituted with cyano, halo, hydroxy, carboxy,
C.sub.1-4 alkylsulfonyl, or C.sub.1-4 alkoxy; and R.sup.2b is H. In
some such embodiments, R.sup.2a is --OR, --O(CH.sub.2).sub.1-3OR,
or --NRC(O)OR, where each R is independently C.sub.1-4 alkyl
optionally substituted with cyano, halo, hydroxy, carboxy,
C.sub.1-4alkylsulfonyl, or C.sub.1-4alkoxy; and R.sup.2b is H In
some preferred embodiments, R.sup.2a is OMe. In these compounds,
R.sup.1 can be H, Me, 4-tetrahydropyranyl, or 2-hydroxy-2-propyl.
In specific embodiments, R.sup.2a is --OMe, --OCH.sub.2CH.sub.2CN,
--OCH.sub.2CH.sub.2OMe, or --OCH.sub.2CH.sub.2SO.sub.2Me or
--NHCOOMe, or --N(Me)COOMe. Preferably when R.sup.2b is H, R.sup.1
is not H.
[0133] Each of the species in Tables 1 and 2 are preferred
embodiments of the invention.
[0134] For purposes of the present invention, a therapeutically
effective dose will generally be a total daily dose administered to
a host in single or divided doses may be in amounts, for example,
of from 0.001 to 1000 mg/kg body weight daily, typically 0.01 to 10
mg/kg per day, and more preferred from 0.1 to 30 mg/kg body weight
daily. Generally, daily dosage amounts of 1 to 4000 mg, or from 5
to 3000, or from 10 to 2000 mg, or from 100 to 2000 mg are
anticipated for human subjects. Dosage unit compositions may
contain such amounts of submultiples thereof to make up the daily
dose.
[0135] The compounds of the present invention may be administered
orally, parenterally, sublingually, by aerosolization or inhalation
spray, rectally, or topically in dosage unit formulations
containing conventional nontoxic pharmaceutically acceptable
carriers, adjuvants, and vehicles as desired. Topical
administration may also involve the use of transdermal
administration such as transdermal patches or ionophoresis devices.
The term parenteral as used herein includes subcutaneous
injections, intravenous, intramuscular, intrasternal injection, or
infusion techniques.
[0136] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions may be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution or suspension in a nontoxic
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-propanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution, and
isotonic sodium chloride solution. In addition, sterile, fixed oils
are conventionally employed as a solvent or suspending medium. For
this purpose any bland fixed oil may be employed including
synthetic mono- or di-glycerides. In addition, fatty acids such as
oleic acid find use in the preparation of injectables.
[0137] Suppositories for rectal administration of the drug can be
prepared by mixing the drug with a suitable nonirritating excipient
such as cocoa butter and polyethylene glycols, which are solid at
ordinary temperatures but liquid at the rectal temperature and will
therefore melt in the rectum and release the drug.
[0138] Solid dosage forms for oral administration may include
capsules, tablets, pills, powders, and granules. In such solid
dosage forms, the active compound may be admixed with at least one
inert diluent such as sucrose lactose or starch. Such dosage forms
may also comprise, as is normal practice, additional substances
other than inert diluents, e.g., lubricating agents such as
magnesium stearate. In the case of capsules, tablets, and pills,
the dosage forms may also comprise buffering agents. Tablets and
pills can additionally be prepared with enteric coatings.
[0139] Liquid dosage forms for oral administration may include
pharmaceutically acceptable emulsions, solutions, suspensions,
syrups, and elixirs containing inert diluents commonly used in the
art, such as water. Such compositions may also comprise adjuvants,
such as wetting agents, emulsifying and suspending agents,
cyclodextrins, and sweetening, flavoring, and perfuming agents.
[0140] The compounds of the present invention can also be
administered in the form of liposomes. As is known in the art,
liposomes are generally derived from phospholipids or other lipid
substances. Liposomes are formed by mono- or multi-lamellar
hydrated liquid crystals that are dispersed in an aqueous medium.
Any non-toxic, physiologically acceptable and metabolizable lipid
capable of forming liposomes can be used. The present compositions
in liposome form can contain, in addition to a compound of the
present invention, stabilizers, preservatives, excipients, and the
like. The preferred lipids are the phospholipids and phosphatidyl
cholines (lecithins), both natural and synthetic. Methods to form
liposomes are known in the art. See, for example, Prescott, Ed.,
Methods in Cell Biology, Volume XIV, Academic Press, New York,
N.W., p. 33 et seq. (1976).
[0141] While the compounds of the invention can be administered as
the sole active pharmaceutical agent, they can also be used in
combination with one or more other agents used in the treatment of
cancer. The compounds of the present invention are also useful in
combination with known therapeutic agents and anti-cancer agents,
and combinations of the presently disclosed compounds with other
anti-cancer or chemotherapeutic agents are within the scope of the
invention. Examples of such agents can be found in Cancer
Principles and Practice of Oncology, V. T. Devita and S. Hellman
(editors), 6.sup.th edition (Feb. 15, 2001), Lippincott Williams
& Wilkins Publishers. A person of ordinary skill in the art
would be able to discern which combinations of agents would be
useful based on the particular characteristics of the drugs and the
cancer involved. Such anti-cancer agents include, but are not
limited to, the following: MEK inhibitors, estrogen receptor
modulators, androgen receptor modulators, retinoid receptor
modulators, cytotoxic/cytostatic agents, antiproliferative agents,
prenyl-protein transferase inhibitors, HMG-CoA reductase inhibitors
and other angiogenesis inhibitors, inhibitors of cell proliferation
and survival signaling, apoptosis inducing agents and agents that
interfere with cell cycle checkpoints. The compounds of the
invention are also useful when co-administered with radiation
therapy.
[0142] Therefore, in one embodiment of the invention, the compounds
of the invention are also used in combination with known
therapeutic or anticancer agents including, for example, estrogen
receptor modulators, androgen receptor modulators, retinoid
receptor modulators, cytotoxic agents, antiproliferative agents,
prenyl-protein transferase inhibitors, HMG-CoA reductase
inhibitors, HIV protease inhibitors, reverse transcriptase
inhibitors, and other angiogenesis inhibitors.
[0143] In certain presently preferred embodiments of the invention,
representative therapeutic agents useful in combination with the
compounds of the invention for the treatment of cancer include, for
example, MEK inhibitors, irinotecan, topotecan, gemcitabine,
5-fluorouracil, cytarabine, daunorubicin, PI3 Kinase inhibitors,
mTOR inhibitors, DNA synthesis inhibitors, leucovorin carboplatin,
cisplatin, taxanes, tezacitabine, cyclophosphamide, vinca
alkaloids, imatinib (Gleevec), anthracyclines, rituximab,
trastuzumab, Revlimid, Velcade, dexamethasone, daunorubicin,
cytaribine, clofarabine, Mylotarg, lenalidomide, bortezomib, as
well as other cancer chemotherapeutic agents including targeted
therapuetics.
[0144] The above compounds to be employed in combination with the
compounds of the invention will be used in therapeutic amounts as
indicated in the Physicians' Desk Reference (PDR) 47th Edition
(1993), which is incorporated herein by reference, or such
therapeutically useful amounts as would be known to one of ordinary
skill in the art, or provided in prescribing materials such as a
drug label for the additional therapeutic agent.
[0145] The compounds of the invention and the other anticancer
agents can be administered at the recommended maximum clinical
dosage or at lower doses. Dosage levels of the active compounds in
the compositions of the invention may be varied so as to obtain a
desired therapeutic response depending on the route of
administration, severity of the disease and the response of the
patient. The combination can be administered as separate
compositions or as a single dosage form containing both agents.
When administered as a combination, the therapeutic agents can be
formulated as separate compositions, which are given at the same
time or different times, or the therapeutic agents, can be given as
a single composition.
[0146] In one embodiment, the invention provides a method of
inhibiting Pim1, Pim2 or Pim3 in a human or animal subject. The
method includes administering an effective amount of a compound, or
a pharmaceutically acceptable salt thereof, of any of the
embodiments of compounds of Formula I or II to a subject in need
thereof.
[0147] The present invention will be understood more readily by
reference to the following examples, which are provided by way of
illustration and are not intended to be limiting of the present
invention.
Synthetic Methods
[0148] The compounds of the invention can be obtained through
procedures known to those skilled in the art. As shown in Scheme
1,5-alkyl, 4-hydroxy, 3-aminopiperidines can be prepared and
modified to yield 5-alkyl, 4-substituted, 3-aminopiperidinyl
pyridine amides VI as follows. Reaction of Garner's aldehyde with
(R)-4-benzyl-3-propionyloxazolidin-2-one followed by TBS protection
of the resulting alcohol affords compound I. Reduction of the
oxazolidinone followed by introduction of the azide group yields
intermediate II. Deprotection under acidic conditions reveals the
corresponding amino alcohol, which upon protection with the Boc
group followed by mesylation of the primary alcohol yields
intermediate III. Reduction of the azide affords formation of the
piperidine which is subsequently reacted with
4-chloro-3-nitropyridine and following nitro reduction pyridyl
aniline IV is obtained. Aniline IV can be coupled with heterocyclic
acids, which after silyl group deprotection (Va) and modification
of the hydroxyl (activation as mesylate, displacement and
potentially further modification of the displaced group) and Boc
deprotection can afford target amides VI. Alternatively, aniline IV
can be Boc protected, silyl deprotected (Vb), modified at the
hydroxyl position and after aniline Boc deprotection (Vc), amide
coupling and Boc deprotection, 5-alkyl, 4-substituted,
3-aminopiperidinyl pyridine amides VI can be obtained.
##STR00015##
[0149] In Scheme 2, synthetic methods to prepare tetrasubstituted
aminocyclohexylpyridyl amides X are depicted. Methyl
cyclohexanedione can be converted via the monotriflate to the
corresponding cyclohexenoneboronate ester which can undergo
palladium mediated carbon bond formation with 4-chloro, 3-nitro
pyridine to yield nitropyridine substituted cyclohexenone VII.
Ketone reduction followed by dehydration yields a cyclohexadiene
which upon epoxidation (via bromohydrin formation and HBr
elimination), azide epoxide opening, azide reduction and amine Boc
protection yields cyclohexenyl Boc amino alcohol nitro pyridyl
compound VIII. Nitro pyridyl VIII can be converted to the trans
protected amino hydroxy aniline IXa by alcohol protection and
alkene and nitro reduction. Alternatively, the alcohol moiety of
nitropyridyl VIII can be inverted via a mesylation, cyclization,
Boc protection and hydrogenation sequence to provide the all cis
substituted cyclohexyl pyridyl aniline IXb, where the cis hydoxy is
protected in the form of a cylic carbamate. As described above in
Scheme 1 for the preparation of substitiuted piperidine compounds
of the invention, upon amide coupling of the cyclohexyl pyridyl
anilines IXa or IXb to heterocyclic acids and subsequent hydroxyl
and amine deprotection, substituted cyclohexyl compounds of the
invention Xa and Xb can be prepared.
##STR00016## ##STR00017## ##STR00018## ##STR00019##
[0150] In Scheme 3, additional synthetic methods to prepare
tetrasubstituted aminocyclohexylpyridyl amides X are depicted in
which the cyclohexyl systems are modified prior to coupling of the
aniline to heterocyclic acids. For example, aniline IXa can be Boc
protected, acetyl deprotected, modified at the hydroxyl position
and after a net aniline Boc deprotection, amide coupling and Boc
deprotection, substituted aminocyclohexylpyridyl amides Xa can be
obtained. In an alternative manner, the cyclohexenyl Boc amino
alcohol nitro pyridyl compound VIII can be modified at the hydroxyl
position and after alkene and nitro reduction, amide coupling and
Boc deprotection, substituted aminocyclohexylpyridyl amides Xc can
be obtained. Additionally, the hydroxyl group of VIII can be
inverted via mesylation, intramolecular cyclization, Boc protection
and cyclic carbamate opening to yield an alcohol that after
processing as described above can yield substituted
aminocyclohexylpyridyl amides Xd. For the sequences depicted in
Schemes 1-3, the aliphatic amine is primarily protected as a Boc
derivative. As one skilled in the art would expect, alternative
protecting groups and subsequent deprotection conditions for the
amine and hydroxy moities can be utilized.
##STR00020## ##STR00021## ##STR00022##
[0151] A representative route to a heterocyclic acid that can be
incorporated into compounds VI, Xa-Xd of the invention is depicted
in Scheme 4. Lithiation of 5-methoxy, 1,3 difluorobenzene and
reaction with 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
yields a boranate ester which upon Suzuki coupling with the
bromofluoro picolinate ester and subsequent ester hydrolysis yields
the fluoro picolinic acid XI.
##STR00023##
EXAMPLES
[0152] Referring to the examples that follow, compounds of the
preferred embodiments were synthesized using the methods described
herein, or other methods, which are known in the art.
[0153] The compounds and/or intermediates were characterized by
high performance liquid chromatography (HPLC) using a Waters
Millenium chromatography system with a 2695 Separation Module
(Milford, Mass.). The analytical columns were reversed phase
Phenomenex Luna C18 -5.mu., 4.6.times.50 mm, from Alltech
(Deerfield, Ill.). A gradient elution was used (flow 2.5 mL/min),
typically starting with 5% acetonitrile/95% water and progressing
to 100% acetonitrile over a period of 10 minutes. All solvents
contained 0.1% trifluoroacetic acid (TFA). Compounds were detected
by ultraviolet light (UV) absorption at either 220 or 254 nm. HPLC
solvents were from Burdick and Jackson (Muskegan, Mich.), or Fisher
Scientific (Pittsburgh, Pa.).
[0154] In some instances, purity was assessed by thin layer
chromatography (TLC) using glass or plastic backed silica gel
plates, such as, for example, Baker-Flex Silica Gel 1B2-F flexible
sheets. TLC results were readily detected visually under
ultraviolet light, or by employing well-known iodine vapor and
other various staining techniques.
[0155] Mass spectrometric analysis was performed on one of three
LCMS instruments: a Waters System (Alliance HT HPLC and a Micromass
ZQ mass spectrometer; Column: Eclipse XDB-C18, 2.1.times.50 mm;
gradient: 5-95% (or 35-95%, or 65-95% or 95-95%) acetonitrile in
water with 0.05% TFA over a 4 min period; flow rate 0.8 mL/min;
molecular weight range 200-1500; cone Voltage 20 V; column
temperature 40.degree. C.), another Waters System (ACQUITY UPLC
system and a ZQ 2000 system; Column: ACQUITY UPLC HSS-C18, 1.8 um,
2.1.times.50 mm; gradient: 5-95% (or 35-95%, or 65-95% or 95-95%)
acetonitrile in water with 0.05% TFA over a 1.3 min period; flow
rate 1.2 mL/min; molecular weight range 150-850; cone Voltage 20 V;
column temperature 50.degree. C.) or a Hewlett Packard System
(Series 1100 HPLC; Column: Eclipse XDB-C18, 2.1.times.50 mm;
gradient: 5-95% acetonitrile in water with 0.05% TFA over a 4 min
period; flow rate 0.8 mL/min; molecular weight range 150-850; cone
Voltage 50 V; column temperature 30.degree. C.). All masses were
reported as those of the protonated parent ions.
[0156] Nuclear magnetic resonance (NMR) analysis was performed on
some of the compounds with a Varian 400 MHz NMR (Palo Alto,
Calif.). The spectral reference was either TMS or the known
chemical shift of the solvent.
[0157] Preparative separations are carried out using a Flash 40
chromatography system and KP-Sil, 60A (Biotage, Charlottesville,
Va.), or by flash column chromatography using silica gel (230-400
mesh) packing material on ISCO or Analogix purification systems, or
by HPLC using a Waters 2767 Sample Manager, C-18 reversed phase
column, 30.times.50 mm, flow 75 mL/min. Typical solvents employed
for the Flash 40 Biotage, ISCO or Analogixsystem for silica gel
column chromatography are dichloromethane, methanol, ethyl acetate,
hexane, n-heptanes, acetone, aqueous ammonia (or ammonium
hydroxide), and triethyl amine. Typical solvents employed for the
reverse phase HPLC are varying concentrations of acetonitrile and
water with 0.1% trifluoroacetic acid.
[0158] Chiral separations of enantiomeric mixtures were performed
by the following analytical and preparative general methods:
[0159] Chiral SFC-Analytical Method: Chiral compounds were
separated on a Waters Supercritical Fluid Chromatography (SFC). The
separation used a Chiralpak AD (AS, OD, OJ, IC or IA) 4.6.times.100
mm column at 40 C temperature at a flow rate of 5 mL/min using an
isocratic method. The mobile phase was 15% MeOH (or EtOH or IPA or
with 0.1% Diethyl amine): 85% CO2. The detection wavelength was 220
nm (or 250 nm or Diode Array)
[0160] Chiral SFC-Purification Method: Chiral compounds were
separated on a Waters Supercritical Fluid Chromatography (SFC). The
separation used a Chiralpak AD (AS, OD, OJ, IC or IA) 21.times.250
mm column at 40 C temperature at a flow rate of 100 mL/min using an
isocratic method. The mobile phase was 15% MeOH (or EtOH or IPA or
with 0.1% Diethyl amine): 85% CO2. The detection wavelength was 220
nm (or 250 nm or Diode Array).
[0161] Chiral HPLC-Analytical Method: Chiral compounds were
separated on a Waters 2695 HPLC system. The separation used a
Chiralpak AD (AS, OD, OJ, IC or IA) 4.6.times.100 mm column at room
temperature at a flow rate of 1 mL/min using an isocratic method.
The mobile phase was 15% EtOH (or IPA or with 0.1% Diethyl amine):
85% Heptane. The detection wavelength was 220 nm (or 250 nm or
Diode Array).
[0162] Chiral HPLC-Purification Method: Chiral compounds were
separated on a Waters 2767 HPLC system. The separation used a
Chiralpak AD (AS, OD, OJ, IC or IA) 21.times.250 mm column at room
temperature at a flow rate of 20 (or 10-15) mL/min using an
isocratic method. The mobile phase was 15% EtOH (or IPA or with
0.1% Diethyl amine): 85% Heptane. The detection wavelength was 220
nm (or 250 nm or Diode Array).
[0163] It should be understood that the organic compounds according
to the preferred embodiments may exhibit the phenomenon of
tautomerism. As the chemical structures within this specification
can only represent one of the possible tautomeric forms, it should
be understood that the preferred embodiments encompasses any
tautomeric form of the drawn structure.
[0164] It is understood that the invention is not limited to the
embodiments set forth herein for illustration, but embraces all
such forms thereof as come within the scope of the above
disclosure.
[0165] The examples below as well as throughout the application,
the following abbreviations have the following meanings. If not
defined, the terms have their generally accepted meanings.
TABLE-US-00001 ABBREVIATIONS BINAP
2,2'-Bis(diphenylphosphino)-1,1'-binaphthalene Boc.sub.2O
di-tert-butyl dicarbonate Boc--OSu
N-(tert-Butoxycarbonyloxy)succinimide Cbz--OSu
N-(Benzyloxycarbonyloxy)succinimide DAST
(diethylamino)sulfurtrifluoride DBU
1,8-Diazabicyclo[5.4.0]undec-7-ene DCM Dichloromethane DIAD
diisopropylazodicarboxylate DIEA diisopropylethylamine DMA
Dimethylacetamide DMAP 4-dimethylaminopyridine DME
1,2-dimethoxyethane DMF N,N-dimethylformamide DPPF
1,1'-bis(diphenylphosphino)ferrocene EDC
1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride EtOAc
ethyl acetate EtOH Ethanol HOAT Hydroxyazabenzotriazole
K.sub.2CO.sub.3 Potassium carbonate KSAc Potassium thioacetate
Lawesson's 2,4-bis(4-methoxyphenyl)-1,3,2,4- reagent
dithiadiphosphetane-2,4-dithione LiOH Lithium hydroxide MCPBA
Meta-chloroperbenzoic acid MeCN Acetonitrile methylDAST
(dimethylamino)sulfurtrifluoride MgSO.sub.4 Magnesium sulfate MeOH
Methanol MsCl Methane sulfonyl chloride Na.sub.2CO.sub.3 sodium
carbonate NaCl Sodium chloride NaHCO.sub.3 sodium bicarbonate
NaHMDS Sodium bis(trimethylsilyl)amide NBS N-bromosuccinimide NMP
N-methyl-2-pyrrolidone oxone Potassium peroxymonosulfate p-TSA
para-toluene sulfonic acid Pd.sub.2(dba).sub.3
Tris(dibenzylideneacetone)dipalladium(0) Pd(PPh.sub.3).sub.4
Tetrakis(triphenylphospine)palladium(0) Pd(dppf)Cl.sub.2--
Dichloro-(1,2-bis(diphenylphosphino)ethan)- DCM Palladium(II) -
dichloromothethane adduct RT or rt room temperature TBAF Tetrabutyl
ammonium fluoride TBDMSCl tert-butyldimethylsilylchloride TEA
Triethylamine THF tetrahydrofuran
EXAMPLES
Synthesis of 6-bromo-5-fluoropicolinic acid
##STR00024##
[0167] To 2-bromo-3-fluoro-6-methylpyridine (1.0 equiv.) in
H.sub.2O (30 mL) was added potassium permanganate (1.0 equiv.). The
solution was heated at 100.degree. C. for 5 hours at which time
more potassium permanganate (1.0 equiv.) was added. After heating
for an additional 48 hours the material was filtered through celite
(4 cm.times.2 inches) and rinsed with H.sub.2O (150 mL). The
combined aqueous was acidified with 1N HCl to pH=4, extracted with
ethyl acetate (200 mL), washed with NaCl(sat.), dried over
MgSO.sub.4, filtered and concentrated to yield
6-bromo-5-fluoropicolinic acid (17%) as a white solid. LCMS (m/z):
221.9 (MH+); LC Rt=2.05 min.
Synthesis of methyl 6-bromo-5-fluoropicolinate
##STR00025##
[0169] To a solution of 6-bromo-5-fluoropicolinic acid (1.0 equiv.)
in methanol (0.2 M) was added H.sub.2SO.sub.4 (4.2 equiv.) and the
reaction was stirred at room temperature for two hours. Upon
completion of the reaction as monitored by LC/MS, the reaction was
diluted with ethyl acetate and quenched slowly with saturated
aqueous NaHCO.sub.3. The reaction was poured into a separatory
funnel and extracted with ethyl acetate. The organic phase was
dried with magnesium sulfate, filtered, and concentrated in vacuo
to provide methyl 6-bromo-5-fluoropicolinate as a white solid
(>99%). LC/MS=233.9/235.9 (M+H), Rt=0.69 min.
Method 1
Synthesis of methyl
6-(2,6-difluoro-4-methoxyphenyl)-5-fluoropicolinate
##STR00026##
[0171] To a solution of methyl 6-bromo-5-fluoropicolinate (1.0
equiv.) in THF and water (10:1, 0.1 M) was added
2,6-difluoro-4-methoxyphenylboronic acid (2.5 equiv.) and potassium
fluoride (3.3 equiv.). The reaction was degassed with nitrogen,
then Pd.sub.2(dba).sub.3 (0.25 equiv.) and tri-tert-butylphosphine
(0.5 equiv.) were added and the reaction was heated to 80.degree.
C. for one hour. LC/MS analysis indicated complete conversion of
the starting material to product. The reaction was cooled to room
temperature, then concentrated in vacuo and fused to silica gel.
The crude product was purified by ISCO flash chromatography eluting
with ethyl acetate and hexanes (0% to 30% ethyl acetate) to provide
methyl 6-(2,6-difluoro-4-methoxyphenyl)-5-fluoropicolinate as a
white solid in 85% yield. LC/MS=298.0 (M+H), Rt=0.89 min.
Method 2
Synthesis of 6-(2,6-difluoro-4-methoxyphenyl)-5-fluoropicolinic
acid
##STR00027##
[0173] To a solution of methyl
6-(2,6-difluoro-4-methoxyphenyl)-5-fluoropicolinate (1.0 equiv.) in
THF/MeOH (2:1, 0.09 M) was added LiOH (1.5 equiv.) and the reaction
was stirred at room temperature for 1 hour. The solution was
quenched with 1N HCl, extracted with ethyl acetate, washed with
brine, dried with sodium sulfate, filtered and concentrated to give
6-(2,6-difluoro-4-methoxyphenyl)-5-fluoropicolinic acid in 84%
yield. LC/MS=284.1 (M+H), Rt=0.76 min.
Method 3
Synthesis of
2-(2,6-difluoro-4-methylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaboroane
##STR00028##
[0175] To a solution of 1,3-difluoro-5-methylbenzene (1.0 eq) in
dry THF (0.2M) under an atmosphere of N.sub.2 at -78.degree. C. was
added n-butyllithium (1 eq, 1.6M in hexanes) slowly keeping the
internal temperature below -65.degree. C. The reaction was stirred
for 2 hrs at -78.degree. C., followed by the addition of
2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.15 eq). The
reaction was allowed to warm to room temperature. Upon completion,
the reaction was quenched with NaHCO.sub.3 (sat.) and extracted
with EtOAc. The organics were washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated to yield
2-(2,6-difluoro-4-methylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaboroane
as a white solid in 92%. 1H NMR (400 MHz, <cdcl3>) .delta.
ppm 6.67 (dd, J=9.39, 0.78 Hz, 2H), 2.34 (s, 3H), 1.38 (s,
12H).
Synthesis of methyl
6-(2,6-difluoro-4-methylphenyl)-5-fluoropicolinate
##STR00029##
[0177] Method 1 was followed using methyl
6-bromo-5-fluoropicolinate (1.0 equiv.) and
2-(2,6-difluoro-4-methylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaboroane
(1.75 equiv.) to give methyl
6-(2,6-difluoro-4-methylphenyl)-5-fluoropicolinate as a solid in
85% yield. LC/MS=282.0 (M+H), Rt=0.87 min.
Synthesis of 6-(2,6-difluoro-4-methylphenyl)-5-fluoropicolinic
acid
##STR00030##
[0179] To a solution of methyl
6-(2,6-difluoro-4-methylphenyl)-5-fluoropicolinate (1.0 eq) in THF
(0.1M) was added LiOH (5.5 eq, 2M) and allowed to stir at room
temperature for 4 hrs. The volatiles were removed in vacuo, and the
residual aqueous was acidified with 2M HCl to pH 4. The precipitate
was filtered and dried to yield
6-(2,6-difluoro-4-methylphenyl)-5-fluoropicolinic acid as a light
yellow solid in 73.5%. LCMS (m/z): 268.0 (MH), R.sub.t=0.76
min.
Synthesis of
(2-(3,5-difluorophenyl)propan-2-yloxy)triisopropylsilane
##STR00031##
[0181] To a solution of 1-(3,5-difluorophenyl)ethanone (1.0 equiv)
in THF (0.2 M) at 0.degree. C. was added methylmagnesium bromide
(1.0 M in THF, 1.15 equiv). After stirring for 4 hours the reaction
was quenched by addition of NH.sub.4Cl.sub.(sat.), diluted with
EtOAc, washed with NaCl.sub.(sat.), dried over MgSO.sub.4,
filtered, concentrated and purified by ISCO SiO.sub.2
chromatography to yield 2-(3,5-difluorophenyl)propan-2-ol. To a
solution of 2-(3,5-difluorophenyl)propan-2-ol in CH.sub.2Cl.sub.2
(0.1 M) at 0.degree. C. was added 2,6 lutidine (6 equiv.) and than
triisopropylsilyl trifluoromethanesulfonate (3.0 equiv.). After
stirring for 3 hours at 0.degree. C. and six hours at rt the
solution was partitioned between EtOAc and NaHCO.sub.3(sat.),
separated, washed with NaCl.sub.(sat.), dried over MgSO.sub.4,
filtered, concentrated and purified by ISCO SiO.sub.2
chromatography to yield
(2-(3,5-difluorophenyl)propan-2-yloxy)triisopropylsilane. (400 MHz,
<cdcl3>) .delta. ppm 1.05-1.08 (m, 21H) 1.57 (s, 6H) 6.63 (s,
1H) 7.00 (dd, J=9.39, 2.35 Hz, 2H).
Synthesis of
(2-(3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)pr-
opan-2-yloxy)triisopropylsilane
##STR00032##
[0183] To a solution of
(2-(3,5-difluorophenyl)propan-2-yloxy)triisopropylsilane (1.0 eq)
in dry THF (0.2M) under an atmosphere of N.sub.2 at -78.degree. C.
was added n-butyllithium (1 eq, 1.6M in hexanes) slowly keeping the
internal temperature below -65.degree. C. The reaction was stirred
for 2 hrs at -78.degree. C., followed by the addition of
2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.15 eq). The
reaction was allowed to warm to room temperature. Upon completion,
the reaction was quenched with NaHCO.sub.3 (sat.) and extracted
with EtOAc. The organics were washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated to yield
(2-(3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)pr-
opan-2-yloxy)triisopropylsilane in 99%. .sup.1H NMR (400 MHz,
<cdcl3>) .delta. ppm 1.03-1.08 (m, 21H) 1.24 (s, 12H) 1.38
(s, 3H) 1.57 (s, 3H) 6.92-7.03 (m, 2H).
Synthesis of tert-butyl(3,5-difluorophenoxy)dimethylsilane
##STR00033##
[0185] To a solution of 3,5-difluorophenol (1.0 equiv.) and
imidazole (2.2 equiv.) in DMF (0.8 M) at 0.degree. C. was added
TBDMSCl (1.1 equiv.). The ice bath was removed and after stirring
for 3 hours the solution was diluted with EtOAc, washed with water,
brine, dried over MgSO.sub.4, filtered, concentrated and purified
by SiO.sub.2 chromatography to yield
tert-butyl(3,5-difluorophenoxy)dimethylsilane in 73% yield. .sup.1H
NMR (400 MHz, <cdcl3>) .delta. ppm 0.23 (s, 6H) 0.99 (s, 9H)
6.33-6.40 (m, 2H) 6.44 (tt 1H).
Synthesis of
tert-butyl(3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ph-
enoxy)dimethylsilane
##STR00034##
[0187] To a solution of
tert-butyl(3,5-difluorophenoxy)dimethylsilane (1.0 eq) in dry THF
(0.2M) under an atmosphere of N.sub.2 at -78.degree. C. was added
n-butyllithium (1 eq, 1.6M in hexanes) slowly keeping the internal
temperature below -65.degree. C. The reaction was stirred for 1 hr
at -78.degree. C., followed by the addition of
2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.1 eq). The
reaction was allowed to warm to room temperature. Upon completion,
the reaction was quenched with NaHCO.sub.3 (sat.) and extracted
with EtOAc. The organics were washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated to yield
tert-butyl(3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ph-
enoxy)dimethylsilane in 91% yield. .sup.1H NMR (400 MHz,
<cdcl3>) .delta. ppm 0.21 (s, 6H) 0.97 (s, 9H) 1.37 (s, 12H)
6.33 (d, J=9.39 Hz, 2H).
Synthesis of methyl
6-(2,6-difluoro-4-hydroxyphenyl)-5-fluoropicolinate
##STR00035##
[0189] Method 1 was followed using methyl
6-bromo-5-fluoropicolinate (1.0 equiv.) and
tert-butyl(3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ph-
enoxy)dimethylsilane (1.75 equiv.) to give methyl
6-(2,6-difluoro-4-hydroxyphenyl)-5-fluoropicolinate in 65% yield.
The reaction was heated for an additional 30 minutes at 100.degree.
C. in the microwave to drive to completion the deprotection of the
TBDMS group. LC/MS=283.9 (M+H), Rt=0.69 min.
Synthesis of methyl
6-(4-(2-(tert-butyldimethylsilyloxy)ethoxy)-2,6-difluorophenyl)-5-fluorop-
icolinate
##STR00036##
[0191] To a solution of methyl
6-(2,6-difluoro-4-hydroxyphenyl)-5-fluoropicolinate (1.0 equiv.)
and potassium carbonate (4.0 equiv.) in DMF (0.4 M) was added
(2-bromoethoxy)(tert-butyl)dimethylsilane (2 equiv.). After
stirring for 72 hours at rt the heterogeneous solution was diluted
with water, extracted with EtOAc, dried over MgSO.sub.4, filtered,
concentrated and purified by ISCO SiO.sub.2 chromatography to yield
methyl
6-(4-(2-(tert-butyldimethylsilyloxy)ethoxy)-2,6-difluorophenyl)-5-fluorop-
icolinate in 74% yield. LC/MS=442.1 (M+H), R.sub.t=1.22 min.
Synthesis of
6-(4-(2-(tert-butyldimethylsilyloxy)ethoxy)-2,6-difluorophenyl)-5-fluorop-
icolinic acid
##STR00037##
[0193] Method 2 was followed using methyl
6-(4-(2-(tert-butyldimethylsilyloxy)ethoxy)-2,6-difluorophenyl)-5-fluorop-
icolinate to give
6-(4-(2-(tert-butyldimethylsilyloxy)ethoxy)-2,6-difluorophenyl)-5-fluorop-
icolinic acid in 94% yield. LC/MS=428.1 (M+H), R.sub.t=1.13
min.
Synthesis of 1,3-difluoro-5-(2-methoxyethoxyl)benzene
##STR00038##
[0195] To a solution of 3,5-difluorophenol (1.0 equiv.),
2-methoxyethanol (3.0 equiv.) and triphenylphosphine (3.0 equiv) in
THF (0.1 M) was added DIAD (3.0 equiv.). After stirring at rt for
18 hours, the volatiles were removed in vacuo and the residue was
purified by SiO.sub.2 chromatography to yield
1,3-difluoro-5-(2-methoxyethoxyl)benzene in 95% yield. .sup.1H NMR
(400 MHz, <cdcl3>) .delta. ppm 6.41-6.47 m (3H), 4.08 (t,
2H), 3.74 (t, 2H), 3.45 (s, 3H).
Synthesis of
2-(2,6-difluoro-4-(2-methoxyethoxyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dio-
xaborolane
##STR00039##
[0197] To a solution of 1,3-difluoro-5-(2-methoxyethoxyl)benzene
(1.0 eq) in dry THF (0.2M) under an atmosphere of N.sub.2 at
-78.degree. C. was added n-butyllithium (1 eq, 1.6M in hexanes)
slowly keeping the internal temperature below -65.degree. C. The
reaction was stirred for 1 hr at -78.degree. C., followed by the
addition of 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
(2.1 eq). The reaction was allowed to warm to room temperature.
Upon completion, the reaction was quenched with NaHCO.sub.3 (sat.)
and extracted with EtOAc. The organics were washed with brine,
dried over Na.sub.2SO.sub.4, filtered and concentrated to yield
2-(2,6-difluoro-4-(2-methoxyethoxyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dio-
xaborolane. .sup.1H NMR (400 MHz, <cdcl3>) ppm 6.42 (d, 2H),
4.10 (m, 2H), 3.74 (m, 2H), 3.44 (s, 3H), 1.37 (s, 12H).
Synthesis of methyl
6-(2,6-difluoro-4-(2-methoxyethoxyl)phenyl)-5-fluoropicolinate
##STR00040##
[0199] Method 1 was followed using methyl
6-bromo-5-fluoropicolinate (1.0 equiv.) and
2-(2,6-difluoro-4-(2-methoxyethoxyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dio-
xaborolane (1.75 equiv.) at 80.degree. C. for 1 hour to give methyl
6-(2,6-difluoro-4-(2-methoxyethoxyl)phenyl)-5-fluoropicolinate in
95% yield. LC/MS=341.9 (M+H), R.sub.t=0.89 min.
Synthesis of
6-(2,6-difluoro-4-(2-methoxyethoxyl)phenyl)-5-fluoropicolinic
acid
##STR00041##
[0201] Method 2 was followed using methyl
6-(2,6-difluoro-4-(2-methoxyethoxyl)phenyl)-5-fluoropicolinate to
give 6-(2,6-difluoro-4-(2-methoxyethoxyl)phenyl)-5-fluoropicolinic
acid in 98% yield. LC/MS=327.9 (M+H), R.sub.t=0.71 min.
Synthesis of methyl
6-(2,6-difluoro-4-(2-hydroxypropan-2-yl)phenyl)-5-fluoropicolinate
##STR00042##
[0203] Method 1 was followed using methyl
6-bromo-5-fluoropicolinate (1.0 equiv.) and
(2-(3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)pr-
opan-2-yloxy)triisopropylsilane (1.6 equiv.) at 100.degree. C. for
30 min in the microwave to give methyl
6-(2,6-difluoro-4-(2-hydroxypropan-2-yl)phenyl)-5-fluoropicolinate
in 90% yield. LC/MS=325.9 (MH.sup.+), R.sub.t=0.81 min. 1H NMR (400
MHz, <cdcl3>) .delta. ppm 1.59 (s, 6H), 4.00 (s, 3H), 7.15
(d, J=9.00 Hz, 2H), 7.62-7.68 (m, 1H), 8.23-8.29 (m, 1H).
Synthesis of
6-(2,6-difluoro-4-(2-hydroxypropan-2-yl)phenyl)-5-fluoropicolinic
acid
##STR00043##
[0205] Method 2 was followed using methyl
6-(2,6-difluoro-4-(2-hydroxypropan-2-yl)phenyl)-5-fluoropicolinate
to give
6-(2,6-difluoro-4-(2-hydroxypropan-2-yl)phenyl)-5-fluoropicolinic
acid in 94% yield. LC/MS=312.0 (MH.sup.+), R.sub.t=0.69 min.
Synthesis of 4-(3,5-difluorophenyl)tetrahydro-2H-pyran-4-ol
##STR00044##
[0207] To a solution of 1-bromo-3,5-difluorobenzene (1.6 equiv.) in
THF (0.26 M) under Ar was added Mg turnings (1.6 equiv.). A reflux
condenser was attached and the solution was submerged in a
90.degree. C. oil bath and refluxed for two hours. The
dihydro-2H-pyran-4(3H)-one (1.0 equiv.) was added in THF via
syringe. The solution was left stirring at rt under Ar for 5 hrs.
The reaction solution was quenched by addition of
NH.sub.4Cl.sub.(sat) and the solution was extracted with EtOAc,
washed with NaCl.sub.(sat.), dried over MgSO.sub.4, filtered,
concentrated and purified by ISCO SiO.sub.2 chromatography (0-100%
EtOAc/n-heptanes gradient) to yield
4-(3,5-difluorophenyl)tetrahydro-2H-pyran-4-ol in 71% yield.
.sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 1.59-1.68 (m, 3H),
2.07-2.19 (m, 2H), 3.87-3.93 (m, 4H), 6.72 (tt, J=8.75, 2.20 Hz,
1H), 6.97-7.06 (m, 2H).
Synthesis of 4-(3,5-difluorophenyl)-3,6-dihydro-2H-pyran
##STR00045##
[0209] 4-(3,5-difluorophenyl)tetrahydro-2H-pyran-4-ol (1.0 equiv.)
was dissolved in DCM (0.2 M) and cooled to 0.degree. C. TEA (2.8
equiv.) was added to the solution, followed by MsC1 (1.3 equiv.).
The reaction was stirred at rt for 2 hrs. The solution was cooled
to 0.degree. C. and DBU (3.0 equiv.) was added. The reaction was
stirred at rt for 18 hrs. The solution was concentrated and the
residue was purified by SiO.sub.2 chromatography (0-100% EtOAc in
Heptanes) to afford 4-(3,5-difluorophenyl)-3,6-dihydro-2H-pyran in
38% yield. .sup.1H NMR (400 MHz, <cdcl3>) .delta. ppm
2.42-2.49 (m, 2H), 3.93 (t, J=5.48 Hz, 2H), 4.32 (q, J=2.74 Hz,
2H), 6.16-6.22 (m, 1H), 6.70 (tt, J=8.80, 2.35 Hz, 1H), 6.85-6.94
(m, 2H).
Synthesis of 4-(3,5-difluorophenyl)tetrahydro-2H-pyran
##STR00046##
[0211] To a solution of 4-(3,5-difluorophenyl)-3,6-dihydro-2H-pyran
(1.0 equiv.) in methanol (0.2 M) was added 10% Pd/C (0.05 equiv.).
The reaction was placed under an atmosphere of hydrogen and stirred
for 18 hours. Upon completion, the solution was filtered over a pad
of Celite, the pad was washed with DCM, the filtrate was
concentrated in vacuo to give
4-(3,5-difluorophenyl)tetrahydro-2H-pyran in 71% yield. .sup.1H NMR
(400 MHz, <cdcl3>) .delta. ppm 1.76 (br. s., 4H), 2.75 (br.
s., 1H), 3.50 (br. s., 2H), 4.08 (d, J=9.78 Hz, 2H), 6.56-6.94 (m,
3H).
Synthesis of
2-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)-4,4,5,5-tetramethyl-1-
,3,2-dioxaborolane
##STR00047##
[0213] Method 3 was followed using
2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.2 equiv.),
butyllithium (1.1 equiv.) and
4-(3,5-difluorophenyl)tetrahydro-2H-pyran (1.0 equiv.) to give
2-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)-4,4,5,5-tetramethyl-1-
,3,2-dioxaborolane in 100% yield. .sup.1H NMR (400 MHz,
<cdcl3>) .delta. ppm 1.16-1.19 (m, 12H), 1.65-1.74 (m, 4H),
2.60-2.75 (m, 1H), 3.37-3.51 (m, 2H), 4.01 (dt, J=11.54, 3.42 Hz,
2H), 6.67 (d, J=8.22 Hz, 2H).
Synthesis of methyl
6-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropicolinate
##STR00048##
[0215] Method 1 was followed using methyl
6-bromo-5-fluoropicolinate (1.0 equiv.) and
2-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)-4,4,5,5-tetramethyl-1-
,3,2-dioxaborolane (3.0 equiv.) at 100.degree. C. for 20 min in
microwave to give methyl
6-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropicolinate
in 59% yield. LC/MS=352.2 (MH.sup.+), R.sub.t=0.92 min.
Synthesis of
6-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropicolinic
acid
##STR00049##
[0217] Method 2 was followed using methyl
6-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropicolinate
to give
6-(2,6-difluoro-4-(tetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropicolini-
c acid in 71% yield. LC/MS=338.1 (MH.sup.+), R.sub.t=0.80 min.
Synthesis of
4-(3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)tet-
rahydro-2H-pyran-4-ol
##STR00050##
[0219] Method 3 was followed using
2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.5 equiv.),
butyllithium (2.4 equiv.) and
4-(3,5-difluorophenyl)tetrahydro-2H-pyran-4-ol (1.0 equiv.) to give
4-(3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)tet-
rahydro-2H-pyran-4-ol in 97% yield. .sup.1H NMR (400 MHz,
<cdcl3>) .delta. ppm 1.32-1.42 (m, 12H), 1.56-1.65 (m, 2H),
2.11 (d, J=3.13 Hz, 2H), 3.86-3.92 (m, 4H), 6.99 (d, J=9.00 Hz,
2H).
Synthesis of methyl
6-(2,6-difluoro-4-(4-hydroxytetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropico-
linate
##STR00051##
[0221] Method 1 was followed using methyl
6-bromo-5-fluoropicolinate (1.0 equiv.) and
4-(3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)tet-
rahydro-2H-pyran-4-ol (1.8 equiv.) at to give methyl
6-(2,6-difluoro-4-(4-hydroxytetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropico-
linate in 72% yield. LC/MS=368.0 (MH.sup.+), R.sub.t=0.75 min.
Synthesis of
6-(2,6-difluoro-4-(4-hydroxytetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropico-
linic acid
##STR00052##
[0223] Method 2 was followed using methyl
6-(2,6-difluoro-4-(4-hydroxytetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropico-
linate to give
6-(2,6-difluoro-4-(4-hydroxytetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropico-
linic acid in 69% yield. LC/MS=354.0 (MH.sup.+), R.sub.t=0.64
min.
Synthesis of methyl
6-(2,6-difluoro-4-(4-fluorotetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropicol-
inate
##STR00053##
[0225] To a solution of methyl
6-(2,6-difluoro-4-(4-hydroxytetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropico-
linate (1.0 equiv.) in CH.sub.2Cl.sub.2 (0.04 M) at -78.degree. C.
under Ar was added methylDAST (2.0 equiv.). After addition, the
solution was stirred under Ar at -78.degree. C. for 10 minutes and
then the bath was removed. The reaction was allowed to warm up to
rt and quenched by addition of NaHCO.sub.3(sat.). The solution was
diluted with EtOAc, washed with NaHCO.sub.3(sat.), NaCl.sub.(sat.),
dried over MgSO.sub.4, filtered, concentrated, purified by ISCO
SiO.sub.2 chromatography (0-100 EtOAc/n-heptanes) to yield methyl
6-(2,6-difluoro-4-(4-fluorotetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropicol-
inate in 100% yield. LC/MS=370.0 (MH.sup.+), R.sub.t=0.94 min.
Synthesis of
6-(2,6-difluoro-4-(4-fluorotetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropicol-
inic acid
##STR00054##
[0227] Method 2 was followed using methyl
6-(2,6-difluoro-4-(4-fluorotetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropicol-
inate to give
6-(2,6-difluoro-4-(4-fluorotetrahydro-2H-pyran-4-yl)phenyl)-5-fluoropicol-
inic acid in 95% yield. LC/MS=355.9 (MH.sup.+), R.sub.t=0.81
min.
Synthesis of 1-(3,5-difluorophenyl)cyclobutanol
##STR00055##
[0229] To a solution of 1-bromo-3,5-difluorobenzene (1.0 equiv.) in
THF (0.26 M) under Ar was added Mg turnings (1.6 equiv.). A reflux
condenser was attached and the solution was submerged in a
90.degree. C. oil bath and refluxed for two hours. The
cyclobutanone (1.0 equiv.) was added in THF via syringe. The
solution was left stirring at rt under Ar for 5 hrs. The reaction
solution was quenched by addition of NH.sub.4Cl.sub.(sat) and the
solution was extracted with EtOAc, washed with NaCl.sub.(sat.),
dried over MgSO.sub.4, filtered, concentrated and purified by ISCO
SiO.sub.2 chromatography (0-100% EtOAc/n-heptanes gradient) to
yield 1-(3,5-difluorophenyl)cyclobutanol in 54% yield. .sup.1H NMR
(400 MHz, CHLOROFORM-d) .delta. ppm 1.69-1.83 (m, 1H), 2.03-2.13
(m, 1H), 2.31-2.43 (m, 2H), 2.45-2.56 (m, 2H), 6.71 (tt, J=8.80,
2.35 Hz, 1H), 6.98-7.07 (m, 2H).
Synthesis of
1-(3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)cyc-
lobutanol
##STR00056##
[0231] Method 3 was followed using
2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.5 equiv.),
butyllithium (2.4 equiv.) and 1-(3,5-difluorophenyl)cyclobutanol
(1.0 equiv.) to give
1-(3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)cyc-
lobutanol in 100% yield. .sup.1H NMR (400 MHz, <cdcl3>)
.delta. ppm 1.23-1.25 (m, 12H), 1.69-1.82 (m, 1H), 2.05-2.12 (m,
1H), 2.37 (br. s., 2H), 2.47 (br. s., 2H), 7.00 (d, J=8.80 Hz,
2H).
Synthesis of methyl
6-(2,6-difluoro-4-(1-hydroxycyclobutyl)phenyl)-5-fluoropicolinate
##STR00057##
[0233] Method 1 was followed using methyl
6-bromo-5-fluoropicolinate (1.0 equiv.) and
1-(3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)cyc-
lobutanol (1.6 equiv.) at 100.degree. C. for 30 min in microwave to
give methyl
6-(2,6-difluoro-4-(1-hydroxycyclobutyl)phenyl)-5-fluoropicolinate
in 71% yield. LC/MS=338.0 (MH.sup.+), R.sub.t=0.85 min.
Synthesis of
6-(2,6-difluoro-4-(1-hydroxycyclobutyl)phenyl)-5-fluoropicolinic
acid
##STR00058##
[0235] Method 2 was followed using methyl
6-(2,6-difluoro-4-(1-hydroxycyclobutyl)phenyl)-5-fluoropicolinate
to give
6-(2,6-difluoro-4-(1-hydroxycyclobutyl)phenyl)-5-fluoropicolinic
acid in 90% yield. LC/MS=323.9 (MH.sup.+), R.sub.t=0.74 min.
Synthesis of 4-(3,5-difluorophenoxyl)tetrahydro-2H-pyran
##STR00059##
[0237] To a solution of 3,5-difluorophenol (1.0 equiv.),
tetrahydro-2H-pyran-4-ol (1.2 equiv.), and triphenylphosphine (2.0
equiv.) in THF (0.33 M) at 0.degree. C. was added DIAD (2.0 equiv.)
dropwise. The reaction mixture was stirred at rt overnight. The
mixture was concentrated and purified by flash chromatography over
silica gel (heptanes:ethyl acetate gradient) to give
4-(3,5-difluorophenoxyl)tetrahydro-2H-pyran in 90% yield. .sup.1H
NMR (400 MHz, <cdcl3>) .delta. ppm 1.72-1.84 (m, 2H),
1.96-2.09 (m, 2H), 3.59 (ddd, J=11.64, 8.31, 3.52 Hz, 2H),
3.90-4.04 (m, 2H), 4.44 (tt, J=7.78, 3.77 Hz, 1H), 6.32-6.53 (m,
3H).
Synthesis of
2-(2,6-difluoro-4-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-4,4,5,5-tetramet-
hyl-1,3,2-dioxaborolane
##STR00060##
[0239] Method 3 was followed using
2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.5 equiv.),
butyllithium (1.3 equiv.) and
4-(3,5-difluorophenoxyl)tetrahydro-2H-pyran (1.0 equiv.) to give
2-(2,6-difluoro-4-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-4,4,5,5-tetramet-
hyl-1,3,2-dioxaborolane in 33% yield. .sup.1H NMR (400 MHz,
<cdcl3>) .delta. ppm 1.21-1.34 (m, 12H), 1.78 (dtd, J=12.72,
8.31, 8.31, 3.91 Hz, 2H), 1.93-2.09 (m, 2H), 3.59 (ddd, J=11.64,
8.31, 3.13 Hz, 2H), 3.89-4.01 (m, 2H), 4.48 (tt, J=7.78, 3.77 Hz,
1H), 6.40 (d, J=9.39 Hz, 2H).
Synthesis of methyl
6-(2,6-difluoro-4-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-5-fluoropicolina-
te
##STR00061##
[0241] Method 1 was followed using methyl
6-bromo-5-fluoropicolinate (1.0 equiv.) and
2-(2,6-difluoro-4-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-4,4,5,5-tetramet-
hyl-1,3,2-dioxaborolane (1.5 equiv.) at 100.degree. C. for 30 min
in microwave to give methyl
6-(2,6-difluoro-4-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-5-fluoropicolina-
te in 77% yield. LC/MS=368.0 (MH+), Rt=0.95 min.
Synthesis of
6-(2,6-difluoro-4-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-5-fluoropicolini-
c acid
##STR00062##
[0243] Method 2 was followed using methyl
6-(2,6-difluoro-4-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-5-fluoropicolina-
te to give
6-(2,6-difluoro-4-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-5-fluo-
ropicolinic acid in 100% yield. LC/MS=353.9 (MH+), R.sub.t=0.82
min.
Synthesis of methyl
6-(4-(ethoxymethyl)-2,6-difluorophenyl)-5-fluoropicolinate
##STR00063##
[0245] To a solution of methyl
6-(2,6-difluoro-4-(hydroxymethyl)phenyl)-5-fluoropicolinate (1.0
equiv.) in DMF (0.20 M) (colorless) at 0.degree. C. was added
sodium hydride (1.2 equiv.) and the reaction was stirred at
0.degree. C. for 2 min. Ethyl iodide (1.2 equiv.) was added and the
reaction was allowed to warm to room temperature. After 1 h,
additional 1.0 equiv. of NaH was added and stirred for 15 mi.
Reaction was quenched by the addition of sat. Ammonium chloride.
The aqueous was acidified with conc HCl to pH3 and extracted with
ethyl acetate three times. The organics were combined, dried with
MgSO.sub.4, filtered and concentrated. The crude mixture was used
as is. LC/MS=326.0 (MH.sup.+), R.sub.t=0.94 min.
Synthesis of
6-(4-(ethoxymethyl)-2,6-difluorophenyl)-5-fluoropicolinic acid
##STR00064##
[0247] Method 2 was followed using methyl
6-(4-(ethoxymethyl)-2,6-difluorophenyl)-5-fluoropicolinate to give
6-(4-(ethoxymethyl)-2,6-difluorophenyl)-5-fluoropicolinic acid.
LC/MS=311.9 (MH.sup.+), R.sub.t=0.82 min.
Synthesis of 1,3-difluoro-5-isopropoxybenzene
##STR00065##
[0249] To a solution of 3,5-difluorophenol (1.0 equiv.) in DMF
(0.26 M) was added potassium carbonate (2.2 equiv.) followed by
2-iodopropane (1.1 equiv.) and the reaction was stirred overnight
at room temperature. The reaction was poured into a separatory
funnel and diluted with a 3:1 (v/v) solution of EtOAc:heptanes. The
organic phase was washed with water, then sat'd NaHCO.sub.3. The
remaining organic phase was dried over MgSO.sub.4, filtered and
concentrated in vacuo to provide 1,3-difluoro-5-isopropoxybenzene
in 88% yield. .sup.1H NMR (400 MHz, <cdcl3>) .delta. ppm 1.33
(d, J=6.26 Hz, 6H), 4.48 (dt, J=11.93, 6.16 Hz, 1H), 6.31-6.47 (m,
3H).
Synthesis of
2-(2,6-difluoro-4-isopropoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborola-
ne
##STR00066##
[0251] Method 3 was followed using
2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.2 equiv.),
butyllithium (1.2 equiv.) and 1,3-difluoro-5-isopropoxybenzene (1.0
equiv.) to give
2-(2,6-difluoro-4-isopropoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborola-
ne in 99% yield. .sup.1H NMR (400 MHz, <cdcl3>) .delta. ppm
1.24 (s, 12H), 1.31-1.33 (m, 6H), 4.43-4.56 (m, 1H), 6.31-6.44 (m,
2H).
Synthesis of methyl
6-(2,6-difluoro-4-isopropoxyphenyl)-5-fluoropicolinate
##STR00067##
[0253] Method 1 was followed using methyl
6-bromo-5-fluoropicolinate (0.8 equiv.) and
2-(2,6-difluoro-4-isopropoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborola-
ne (1.0 equiv.) at 70.degree. C. for 1 hour to give methyl
6-(2,6-difluoro-4-isopropoxyphenyl)-5-fluoropicolinate. LC/MS=325.9
(MH+), Rt=1.04 min.
Synthesis of 6-(2,6-difluoro-4-isopropoxyphenyl)-5-fluoropicolinic
acid
##STR00068##
[0255] Method 2 was followed using methyl
6-(2,6-difluoro-4-isopropoxyphenyl)-5-fluoropicolinate to give
6-(2,6-difluoro-4-isopropoxyphenyl)-5-fluoropicolinic acid.
LC/MS=311.9 (MH.sup.+), Rt=0.92 min.
Synthesis of 3-(3,5-difluorophenyl)oxetane
##STR00069##
[0257] 3,5-difluorophenylboronic acid (2.0 equiv.),
(1R,2R)-2-aminocyclohexanol (0.06 equiv.), NaHMDS (2.0 equiv.), and
nickel(II) iodide (0.06 equiv.) were dissolved in 2-propanol (0.35
M). The mixture was degassed with N.sub.2, stirred at rt for 10 min
and then a solution of 3-iodooxetane (1.0 equiv.) in 2-Propanol
(0.70 M) was added. The mixture was sealed and heated at 80.degree.
C. in the microwave for 20 min. The mixture was filtered through
celite, eluting with EtOH and concentrated. The crude residue was
purified by ISCO SiO.sub.2 chromatography eluting with 0-100% EtOAc
in Heptanes to afford 3-(3,5-difluorophenyl)oxetane in 63% yield.
.sup.1H NMR (400 MHz, <cdcl3>) .delta. 6.88-6.96 (m, 2H),
6.72 (tt, J=2.20, 8.95 Hz, 1H), 5.08 (dd, J=6.26, 8.22 Hz, 2H),
4.71 (t, J=6.26 Hz, 2H), 4.14-4.24 (m, 1H).
Synthesis of
2-(2,6-difluoro-4-(oxetan-3-yl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxabor-
olane
##STR00070##
[0259] Method 3 was followed using
2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.3 equiv.),
butyllithium (1.1 equiv.) and 3-(3,5-difluorophenyl)oxetane (1.0
equiv.) to give
2-(2,6-difluoro-4-(oxetan-3-yl)phenyl)-4,4,5,5-tetramethyl-1,3,2--
dioxaborolane. .sup.1H NMR (400 MHz, <cdcl3>) .delta. ppm
6.90 (d, J=8.22 Hz, 2H), 5.07 (dd, J=6.06, 8.41 Hz, 2H), 4.70 (t,
J=6.26 Hz, 2H), 4.13-4.23 (m, 1H), 1.39 (s, 12H).
Synthesis of methyl
6-(2,6-difluoro-4-(oxetan-3-yl)phenyl)-5-fluoropicolinate
##STR00071##
[0261] Method 1 was followed using methyl
6-bromo-5-fluoropicolinate (1.2 equiv.) and
2-(2,6-difluoro-4-(oxetan-3-yl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxabor-
olane (1.0 equiv.) at 80.degree. C. for 15 min in microwave to give
methyl 6-(2,6-difluoro-4-(oxetan-3-yl)phenyl)-5-fluoropicolinate in
47% yield. LC/MS=324.0 (MH+), Rt=0.75 min.
Synthesis of
6-(2,6-difluoro-4-(oxetan-3-yl)phenyl)-5-fluoropicolinic acid
##STR00072##
[0263] Method 2 was followed using methyl
6-(2,6-difluoro-4-(oxetan-3-yl)phenyl)-5-fluoropicolinate to give
6-(2,6-difluoro-4-(oxetan-3-yl)phenyl)-5-fluoropicolinic acid in
71% yield. LC/MS=309.9 (MH+), Rt=0.69 min.
Synthesis of methyl
2',6,6'-trifluoro-4'-(trifluoromethylsulfonyloxy)biphenyl-3-carboxylate
##STR00073##
[0265] To a solution of methyl
2',6,6'-trifluoro-4'-hydroxybiphenyl-3-carboxylate (1.0 equiv.) in
DCM (0.35 M) at 0.degree. C. was added pyridine (1.5 equiv.) and
allowed to stir for 5 mins, followed by the addition of
TriflicAnhydride (1.1 equiv.). The reaction was allowed to stir
warming to RT. The reaction was quenched with NaHCO.sub.3(sat),
extracted in DCM and the organics were washed with water and brine.
The organics were dried over Na.sub.2SO.sub.4, filtered, and
concentrated to yield methyl
2',6,6'-trifluoro-4'-(trifluoromethylsulfonyloxy)biphenyl-3-carboxylate
in 81% yield.
Synthesis of methyl
6-(4-(3,6-dihydro-2H-thiopyran-4-yl)-2,6-difluorophenyl)-5-fluoropicolina-
te
##STR00074##
[0267] To a degassed solution of methyl
6-(2,6-difluoro-4-(trifluoromethylsulfonyloxy)phenyl)-5-fluoropicolinate
(1.0 equiv.) and 3,6-dihydro-2H-thiopyran-4-ylboronic acid (1.5
equiv.) in DME/2M Na.sub.2CO.sub.3 (3/1, 0.10 M) was added
PdCl2(dppf).CH.sub.2Cl.sub.2 adduct (0.10 equiv.). The reaction was
heated to 90.degree. C. in an oil bath for 15 min. The reaction
mixture was partitioned with water and EtOAc; the organics were
dried over MgSO.sub.4, filtered, and concentrated. The crude was
purified via ISCO SiO.sub.2 chromatography. Pure fractions were
combined and concentrated to yield methyl
6-(4-(3,6-dihydro-2H-thiopyran-4-yl)-2,6-difluorophenyl)-5-fluoropicolina-
te in 60% yield. LC/MS=366.1 (M+H), Rt=1.00 min.
Synthesis of methyl
6-(4-(1,1-dioxido-3,6-dihydro-2H-thiopyran-4-yl)-2,6-difluorophenyl)-5-fl-
uoropicolinate
##STR00075##
[0269] To a solution of methyl
6-(4-(3,6-dihydro-2H-thiopyran-4-yl)-2,6-difluorophenyl)-5-fluoropicolina-
te (1.0 equiv.) in DCM (0.10 M) at rt was added oxone (6.0 equiv.)
in one portion. The resulting mixture was stirred at RT overnight,
and then refluxed at 40.degree. C. for 4 hrs. 10.0 equiv. of oxone
were added and the reaction was allowed to stir at 40.degree. C.
over the weekend. The reaction mixture was then diluted with DCM
and washed with water the aqueous layer was then separated and
extracted with DCM. The combined organic were then dried over
MgSO.sub.4 and concentrated in vacuo to yield methyl
6-(4-(1,1-dioxido-3,6-dihydro-2H-thiopyran-4-yl)-2,6-difluorophenyl)-5-fl-
uoropicolinate in 100% yield. LC/MS=398.0 (M+H), Rt=0.76 min.
Synthesis of
6-(4-(1,1-dioxido-3,6-dihydro-2H-thiopyran-4-yl)-2,6-difluorophenyl)-5-fl-
uoropicolinic acid
##STR00076##
[0271] Method 2 was followed using methyl
6-(4-(1,1-dioxido-3,6-dihydro-2H-thiopyran-4-yl)-2,6-difluorophenyl)-5-fl-
uoropicolinate to give
6-(4-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-2,6-difluorophenyl)-5-fluo-
ropicolinic acid in 74% yield. LC/MS=384.0 (M+H), Rt=0.64 min.
Synthesis of
6-(4-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-2,6-difluorophenyl)-5-fluo-
ropicolinic acid
##STR00077##
[0273] To a degassed solution of
6-(4-(1,1-dioxido-3,6-dihydro-2H-thiopyran-4-yl)-2,6-difluorophenyl)-5-fl-
uoropicolinic acid (1.0 equiv.) in EtOH(0.10 M) was added Pd/C (0.1
equiv.). The mixture was stirred at rt under H.sub.2 for 16 hrs.
Add Pd/C (0.1 equiv.) and the reaction was stirred for additional
16 hrs. The reaction was filtered and concentrated to yield
6-(4-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-2,6-difluorophenyl)-5-fluo-
ropicolinic acid in 100% yield. LC/MS=386.0 (M+H), Rt=0.64 min.
Synthesis of methyl
6-(2,6-difluoro-4-(prop-1-en-2-yl)phenyl)-5-fluoropicolinate
##STR00078##
[0275] To a degassed solution of methyl
6-(2,6-difluoro-4-(trifluoromethylsulfonyloxy)phenyl)-5-fluoropicolinate
(1.0 equiv.) in DME/2M Na.sub.2CO.sub.3 (3/1, 0.09 M) was added
4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (1.5
equiv.) and PdCl2(dppf)-CH.sub.2Cl.sub.2Adduct (0.1 equiv.). The
reaction was heated to 90.degree. C. in an oil bath for 15 min. The
mixture was cooled to rt and partitioned between water and ethyl
acetate. The organic phase was dried with sodium sulfate, filtered
and concentrated. The crude material was purified via silica gel
column chromatography (Analogix, eluting with 0-100% ethyl
acetate). The pure fractions were concentrated to yield methyl
6-(2,6-difluoro-4-(prop-1-en-2-yl)phenyl)-5-fluoropicolinate.
LC/MS=308.2 (M+H), Rt=0.99 min. .sup.1H NMR (400 MHz,
<cdcl3>) .delta. ppm 2.15 (s, 3H), 4.01 (s, 3H), 5.23 (s,
1H), 5.47 (s, 1H), 7.11 (d, J=9.39 Hz, 2H), 7.65 (t, J=8.41 Hz,
1H), 8.26 (dd, J=8.61, 3.91 Hz, 1H).
Synthesis of methyl
6-(2,6-difluoro-4-isopropylphenyl)-5-fluoropicolinate
##STR00079##
[0277] To a degassed solution of methyl
6-(2,6-difluoro-4-(prop-1-en-2-yl)phenyl)-5-fluoropicolinate (1.0
equiv.) in MeOH (0.09 M) was added Pd/C (0.1 equiv.) and the
reaction was stirred at rt under an atmosphere of hydrogen. After
overnight stirring, filtered through a pad of Celite and washed
with Methanol. The filtrate was concentrated and dried under vacuo
to give methyl
6-(2,6-difluoro-4-isopropylphenyl)-5-fluoropicolinate. LC/MS=310.0
(M+H), Rt=1.00 min.
Synthesis of 6-(2,6-difluoro-4-isopropylphenyl)-5-fluoropicolinic
acid
##STR00080##
[0279] Method 2 was followed using methyl
6-(2,6-difluoro-4-isopropylphenyl)-5-fluoropicolinate to give
6-(2,6-difluoro-4-isopropylphenyl)-5-fluoropicolinic acid in 100%
yield. LC/MS=296.2 (M+H), Rt=0.89 min.
Synthesis of ethyl 2-(2,6-difluorophenyl)thiazole-4-carboxylate
##STR00081##
[0281] A solution of 2,6-difluorobenzothioamide (1.0 eq) and
ethylbromopyruvate (1.0 eq.) in ethanol (1.0 M) was heated in the
microwave at 130.degree. C. for 30 minutes. Upon removal of
volatiles in vacuo, ethyl acetate was added and the solution was
washed with Na.sub.2CO.sub.3(sat.), with NaCl.sub.(sat.), was dried
over MgSO.sub.4, filtered and concentrated yielding ethyl
2-(2,6-difluorophenyl)thiazole-4-carboxylate (84%). LCMS (m/z):
270.1 (MH.sup.+); LC R.sub.t=3.79 min.
Synthesis of 2-(2,6-difluorophenyl)thiazole-4-carboxylic acid
##STR00082##
[0283] To a solution of ethyl
2-(2,6-difluorophenyl)thiazole-4-carboxylate (1.0 eq.) in 2:1
THF/MeOH (0.17 M) was added 1.0 M LiOH (2.0 eq.). After standing
for 16 hours, 1.0 M HCl (2.0 eq.) was added and the THF/MeOH was
removed in vacuo. The resulting solid was filtered, rinsed with
H.sub.2O and dried, yielding
2-(2,6-difluorophenyl)thiazole-4-carboxylic acid (88%) as a crusty
solid. LCMS (m/z): 251.1 (MH.sup.+); LC R.sub.t=2.68 min.
Synthesis of ethyl 2-amino-2-cyanoacetate
##STR00083##
[0285] To a solution of ethyl 2-cyano-2-(hydroxyimino)acetate(1 eq)
in ethanol (1.4 M) was added PtO.sub.2 (0.05 eq) and the solution
was put under an H.sub.2 atmosphere (4 bar) in a steel bomb and was
stirred overnight. The reaction was filtered through a pad of
celite, rinsing with CH.sub.2Cl.sub.2 and upon removal of voltiles
in vacuo ethyl 2-amino-2-cyanoacetate was obtained in 89% yield.
LC/MS (m/z): 129.0 (MH.sup.+), R.sub.t: 0.25 min.
Synthesis of ethyl 2-cyano-2-(2,6-difluorobenzamido)acetate
##STR00084##
[0287] To a solution of ethyl 2-amino-2-cyanoacetate (1 eq) in 6 mL
of dichloromethane was added pyridine (1.5 eq) and
2,6-difluorobenzoyl chloride (1 eq) at 0.degree. C. The reaction
mixture was stirred at room temperature for 3 hours. The mixture
was diluted with ethyl acetate, washed with brine, then dried over
anhydrous MgSO.sub.4, filtered, and concentrated in vacuo. The
crude residue was purified by flash chromatography (EtOAc:
hexanes=1:1) to give the titled compound (84%). LC/MS (m/z): 269.1
(MH.sup.+), R.sub.t: 0.69 min.
Synthesis of ethyl
5-amino-2-(2,6-difluorophenyl)thiazole-4-carboxylate
##STR00085##
[0289] To a solution of the ethyl
2-cyano-2-(2,6-difluorobenzamido)acetate (1 eq) in pyridine (0.1 M)
was added Lawesson's reagent (1.5 eq.). The mixture was stirred at
reflux under Ar for 18 hours. Solvents were removed under reduced
pressure. The crude residue was purified by flash chromatography
(EtOAc: hexanes=1:1) to give the ethyl
5-amino-2-(2,6-difluorophenyl)thiazole-4-carboxylate in 25% yield.
LC/MS (m/z): 284.9 (MH.sup.+), R.sub.t: 0.76 min.
Synthesis of
5-((tert-butoxycarbonyl)amino)-2-(2,6-difluorophenyl)thiazole-4-carboxyli-
c acid
##STR00086##
[0291] To a solution of the ethyl
5-amino-2-(2,6-difluorophenyl)thiazole-4-carboxylate (1 eq) in
CH.sub.2Cl.sub.2 (0.1 M) was added Boc.sub.2O (1.2 eq.) and DMAP
(0.05 eq.). Upon stirring for 1 hour the volatiles were removed in
vacuo, THF (0.1 M) and 2.0 M LiOH.sub.(aq.) (5 equiv) were added
and the solution was stirred at 55.degree. C. for 2 days. The
volatiles were removed in vacuo and the remaining aqueous solution
was adjusted to pH 5 by addition of 2 M HCl. The resulting solid
was filtered, rinsed with H.sub.2O and pumped on to yield
5-((tert-butoxycarbonyl)amino)-2-(2,6-difluorophenyl)thiazole-4-carboxyli-
c acid 25% yield. LC/MS (m/z): 357.1 (MH.sup.+), R.sub.t: 0.97
min.
Synthesis of Methyl 3-amino-5-fluoropicolinate
##STR00087##
[0293] To a steel bomb reactor, 2-bromo-5-fluoropyridin-3-amine
(1.0 equiv.), triethylamine (1.6 equiv.), Pd(BINAP)Cl.sub.2 (0.0015
equiv.) and anhydrous methanol (0.4 M solution) were added. After
degassed by nitrogen stream for 15 min, the steel bomb reactor was
closed and filled with CO gas up to 60 psi. The reactor was then
heated to 100.degree. C. After 3 h, more Pd catalyst (0.0015
equiv.) was added and the reaction mixture was re-heated to the
same temperature for 3 h. After cooling down to room temperature, a
brown precipitate was filtered off and the filtrate was extracted
with EtOAc, which was washed with water and brine, dried over
anhydrous sodium sulfate, and filtered. After removing volatile
materials, the crude yellow product was obtained and used for the
next step without further purification (40%). LCMS (m/z): 271.2
(MH.sup.+); LC R.sub.t=3.56 min.
Synthesis of Methyl 3-amino-6-bromo-5-fluoropicolinate
##STR00088##
[0295] To a solution of methyl 3-amino-5-fluoropicolinate (1.0
equiv.) in acetonitrile (0.3 M solution) was added NBS (1.1 equiv.)
for 2 minutes at room temperature. After quenched with water, the
reaction mixture was extracted with EtOAc. The crude product was
purified by silica column chromatography (20% to 50% EtOAc in
hexanes) to give methyl 3-amino-6-bromo-5-fluoropicolinate (41%).
LCMS (m/z): 249.1 (MH.sup.+); LC R.sub.t=2.80 min.
Synthesis of methyl
3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinate
##STR00089##
[0297] Method 1 was followed using methyl
3-amino-6-bromo-5-fluoropicolinate (1.0 equiv.) and
2,6-difluorophenylboronic acid (1.3 equiv.) and
Pd(dppf)Cl.sub.2-DCM (0.05 equiv.) to give methyl
3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinate in 94% yield.
LCMS (m/z): 283.0 (MH.sup.+), R.sub.t=0.76 min.
Synthesis of 3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinic
acid
##STR00090##
[0299] Method 2 was followed using methyl
3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinate (1.0 equiv.) and
LiOH (1.0 equiv.) to give
3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinic acid in 79% yield.
LCMS (m/z): 269.0 (MH.sup.+), R.sub.t=0.79 min.
Synthesis of 2-(2,6-difluorophenyl)pyrimidine-4-carboxylic acid
##STR00091##
[0301] To a solution of 2-chloropyrimidine-4-carboxylic acid (1.0
equiv.) in DME and 2M Na.sub.2CO.sub.3 (3:1, 0.25 M) was added
2,6-difluorophenylboronic acid (1.3 equiv.) and
Pd(dppf)Cl.sub.2-DCM (0.05 equiv.) in a microwave vial. The vial
was heated in the microwave at 120.degree. C. for 30 minutes. The
mixture was diluted with ethyl acetate and 1N NaOH was added. The
organic phase was separated and extracted three more times with 1N
NaOH and once with 6N NaOH. The combined aqueous phases were
filtered and acidified to pH 1 by the addition of concentrated HCl
and extracted with ethyl acetate. The organic layer was dried over
magnesium sulfate, filtered, and concentrated to give
2-(2,6-difluorophenyl)pyrimidine-4-carboxylic acid in 81%. LCMS
(m/z): 237.0 (MH.sup.+), R.sub.t=0.54 min.
Synthesis of 5-amino-2-(2,6-difluorophenyl)pyrimidine-4-carboxylic
acid
##STR00092##
[0303] A 2.68 M NaOEt in EtOH solution (3 eq) was added to an
ice-bath cooled mixture of 2, 6-difluorobenzimidamide hydrochloride
(2 eq) in EtOH (0.1 M). The resulting mixture was allowed to warm
to rt and stirred under N.sub.2 for 30 min. To the reaction mixture
was added drop wise a solution of mucobromic acid (1 eq) in EtOH
and the reaction was heated in a 50.degree. C. oil bath for 2.5 hr.
After cooling to rt the reaction mixture was concentrated in vacuo.
H.sub.2O and 1.0 N NaOH were added and the aqueous mixture was
washed with EtOAc. The aqueous phase was acidified to pH=4 with 1.0
N HCl then extracted with EtOAc. Combined organic extracts were
washed once with brine, then dried over anhydrous Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo to give
5-bromo-2-(2,6-difluorophenyl)pyrimidine-4-carboxylic acid. The
crude product was used for the next step without further
purification. LC/MS (m/z): 316.9 (MH.sup.+). LC: R.sub.t: 2.426
min.
[0304] CuSO.sub.4 (0.1 eq) was added to a mixture of
5-bromo-2-(2,6-difluorophenyl)pyrimidine-4-carboxylic acid (1 eq)
and 28% aqueous ammonium hydroxide solution in a microwave reaction
vessel. The reaction mixture was heated in a microwave reactor at
110.degree. C. for 25 min. The reaction vessel was cooled in dry
ice for 30 min then unsealed and concentrated in vacuo. To the
resulting solids was added 1.0 N HCl and the mixture was extracted
with EtOAc. Combined organic extracts were washed once with brine,
then dried over anhydrous Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo to give
5-amino-2-(2,6-difluorophenyl)pyrimidine-4-carboxylic acid. The
crude product was used for the next step without further
purification. LCMS (m/z): 252.0 (MH.sup.+), R.sub.t=2.0 min.
Synthesis of 2-(2,6-difluorophenyl)-3-fluoro-6-methylpyridine
##STR00093##
[0306] To a solution of 2-bromo-3-fluoro-6-methylpyridine (1.0
equiv.) in THF and Water (10:1, 0.2 M) was added
2,6-difluorophenylboronic acid (2.0 equiv.) and potassium fluoride
(3.3 equiv.). The reaction was degassed for 10 minutes, then
Pd.sub.2(dba).sub.3 (0.05 equiv.) was added, followed by
tri-t-butylphosphine (0.1 equiv.). The reaction was stirred to
60.degree. C. for 1 hour at which point, all starting material was
consumed as indicated by LC/MS. The reaction was allowed to cool to
room temperature, partitioned with ethyl acetate and water, the
organic phase was dried with sodium sulfate, filtered, and
concentrated. The crude material was diluted in EtOH to 0.1 M, and
0.5 equiv. of NaBH.sub.4 was added to reduce the dba. The reaction
was stirred for one hour at room temperature, then quenched with
water and concentrated under vacuo to remove the ethanol. The
product was extracted in ether, washed with brine, the organics
were dried over sodium sulfate, filtered, and concentrated. The
crude material was loaded on silica gel and purified via column
chromatography (ISCO) eluting with hexanes and ethyl acetate
(0%-10% ethyl acetate). The pure fractions were combined, and
concentrated to yield
2-(2,6-difluorophenyl)-3-fluoro-6-methylpyridine as a light yellow
oil in 86% yield. LC/MS=224.0 (M+H), R.sub.t=0.84 min.
Synthesis of 6-(2,6-difluorophenyl)-5-fluoropicolinic acid
##STR00094##
[0308] To a solution of
2-(2,6-difluorophenyl)-3-fluoro-6-methylpyridine (1.0 equiv.) in
water (0.05 M) was added KMnO.sub.4 (2.0 equiv.) and the reaction
was heated to reflux overnight. Another 2.0 equiv. of KMnO.sub.4
were added and stirred at reflux for another 8 hours. The solution
was cooled to room temperature, filtered through Celite and washed
with water. The filtrate was acidified with 6N HCl to pH=3, the
white precipitate was filtered. The filtrate was further acidified
to pH=1 and filtered again. The filtrate was extracted with ethyl
acetate until no more product was in the aqueous layer. The organic
phase was washed with brine and dried over magnesium sulfate,
filtered, and concentrated. The residue was dissolved in ethyl
acetate, washed with 1N NaOH, the aqueous layer was acidified to
pH=1 and the white crystals were filtered. The combined products
yielded 6-(2,6-difluorophenyl)-5-fluoropicolinic acid in 32% yield
as a white solid. LC/MS=254.0 (M+H), R.sub.t=0.71 min.
Synthesis of methyl
3-amino-6-(2-fluoro-5-isopropylcabamoyl)phenyl)-picolinate
##STR00095##
[0310] A solution of methyl 3-amino-6-bromopicolinate (1.0 equiv.),
N-isopropyl 3-borono-4-fluorobenzamide (1.1 equiv.), and
Pd(dppf)Cl.sub.2-DCM (0.15 equiv.) in DME/2M Na.sub.2CO.sub.3
(3:1), at a concentration of 0.5 M, was stirred at 120.degree. C.
for 1.5 hours. The reaction was filtered and washed with EtOAc. The
organic was partitioned with H.sub.2O (25 mL), washed with
NaCl.sub.(sat.) (25 mL), dried over MgSO.sub.4, and the volatiles
were removed in vacuo. The residue was diluted in EtOAc and passed
through a silica gel plug and the volatiles were removed in vacuo
yielding methyl
3-amino-6-(2-fluoro-5-isopropylcabamoyl)phenyl)picolinate (60%).
LCMS (m/z): 332.2 (MH.sup.+); LC R.sub.t=2.9 min.
Synthesis of
3-amino-6-(2-fluoro-5-isopropylcabamoyl)phenyl)picolinic acid
##STR00096##
[0312] To a solution of methyl
3-amino-6-(2-fluoro-5-isopropylcabamoyl)phenyl)picolinate (1.0
equiv) in THF (0.5M), was added 1M LiOH (4.0 equiv). After stirring
for 4 hours at 60.degree. C., 1 N HCl (4.0 equiv.) was added and
the THF was removed in vacuo. The resulting solid was filtered and
rinsed with cold H.sub.2O (3.times.20 mL) to yield
3-amino-6-(2-fluoro-5-isopropylcabamoyl)phenyl)picolinic acid
(98%). LCMS (m/z): 318.1 (MH.sup.+); LC R.sub.t=2.4 min.
Synthesis of (R)-tert-butyl
4-((1R,2R)-3-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-hydroxy-2-methyl-3-oxo-
propyl)-2,2-dimethyloxazolidine-3-carboxylate
##STR00097##
[0314] To a solution of (R)-4-benzyl-3-propionyloxazolidin-2-one
(1.0 equiv.) in DCM (0.13 M) was added TiCl.sub.4 (1.0 equiv.) at
-40.degree. C. The mixture was stirred at -40.degree. C. for 10 min
(yellow suspension), then DIPEA (2.5 equiv.) was added (dark red
solution) and stirred at 0.degree. C. for 20 min. (R)-tert-butyl
4-formyl-2,2-dimethyloxazolidine-3-carboxylate (1.0 equiv.) in DCM
(0.5 M) was then added dropwise and the resulting mixture was
stirred for 1.5 hours. The reaction was quenched by the addition of
aqueous ammonium chloride and the mixture was extracted with ethyl
acetate. The organic phase was separated, washed with brine, dried
with magnesium sulfate, filtered, and concentrated. The residue was
purified via column chromatography eluting with ethyl acetate and
hexanes (1:4) to give (R)-tert-butyl
4-((1R,2R)-3-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-hydroxy-2-methyl-3-oxo-
propyl)-2,2-dimethyloxazolidine-3-carboxylate as the major product
(5:2) in 58% yield. LC/MS=363.3 (M+H-Boc), Rt=1.09 min.
Synthesis of (R)-tert-butyl
4-((1R,2R)-3-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(tert-butyldimethylsil-
yloxy)-2-methyl-3-oxopropyl)-2,2-dimethyloxazolidine-3-carboxylate
##STR00098##
[0316] To a solution of (R)-tert-butyl
4-((1R,2R)-3-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-hydroxy-2-methyl-3-oxo-
propyl)-2,2-dimethyloxazolidine-3-carboxylate (1.0 equiv.) and
lutidine (1.8 equiv.) in DCM (0.1M) was added TBSOTf (1.4 equiv.)
at -40.degree. C. The reaction mixture was stirred at -40.degree.
C. for 2 hours. The solution was diluted with ethyl acetate and
washed with sat. NaHCO.sub.3, sat. NaCl, dried with magnesium
sulfate, filtered, and concentrated. The residue was purified by
silica gel column chromatography eluting with ethyl acetate and
hexanes (1:4) to give (R)-tert-butyl
4-((1R,2R)-3-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(tert-butyldimethylsil-
yloxy)-2-methyl-3-oxopropyl)-2,2-dimethyloxazolidine-3-carboxylate
as the major product (5:2) in 83% yield. LC/MS=577.3 (M+H), Rt=1.33
min (Frac 65%-95% method).
Synthesis of (R)-tert-butyl
4-((1R,2S)-1-(tert-butyldimethylsilyloxy)-3-hydroxy-2-methylpropyl)-2,2-d-
imethyloxazolidine-3-carboxylate
##STR00099##
[0318] To a solution of (R)-tert-butyl
4-((1R,2R)-3-((R)-4-benzyl-2-oxooxazolidin-3-yl)-1-(tert-butyldimethylsil-
yloxy)-2-methyl-3-oxopropyl)-2,2-dimethyloxazolidine-3-carboxylate
(1.0 equiv.) and ethanol (3.0 equiv.) in THF (0.09 M) was added
LiBH.sub.4 (3.0 equiv.) at -30.degree. C. The reaction mixture was
allowed to warm up to 0.degree. C. and stirred at that temperature
for 3 hours. The solution was then diluted with diethyl ether and
1N NaOH was added. The resulting mixture was extracted with ethyl
acetate, the organic layer was separated, washed with sat. NaCl,
dried over magnesium sulfate, filtered, and concentrated. The
residue was purified via silica gel column chromatography eluting
with ethyl acetate and hexanes (1:4) to give (R)-tert-butyl
4-((1R,2S)-1-(tert-butyldimethylsilyloxy)-3-hydroxy-2-methylpropyl)-2,2-d-
imethyloxazolidine-3-carboxylate as the major product (5:2 ratio)
in 71% yield. LC/MS=304.3 (M+H-Boc), Rt=0.95 min (Frac 65%-95%
method).
Synthesis of (R)-tert-butyl
4-((1R,2S)-3-azido-1-(tert-butyldimethylsilyloxy)-2-methylpropyl)-2,2-dim-
ethyloxazolidine-3-carboxylate
##STR00100##
[0320] To a solution of (R)-tert-butyl
4-((1R,2S)-1-(tert-butyldimethylsilyloxy)-3-hydroxy-2-methylpropyl)-2,2-d-
imethyloxazolidine-3-carboxylate (1.0 equiv.), DIAD (2.0 equiv.),
and PPh.sub.3(2.0 equiv.) in THF (0.18 M) was added DPPA (2.0
equiv., 1M solution in THF). The reaction mixture was stirred at
room temperature overnight. Upon removal of the volatiles under
vacuo, the residue was purified by silica gel column chromatography
eluting with ethyl acetate and hexanes (1:6) to give (R)-tert-butyl
4-((1R,2S)-3-azido-1-(tert-butyldimethylsilyloxy)-2-methylpropyl)-2,2-dim-
ethyloxazolidine-3-carboxylate as the major product (5:2) in 86%
yield. LC/MS=329.3 (M+H-Boc), Rt=1.40 min (Frac 65%-95%
method).
Synthesis of tert-butyl
(2R,3R,4S)-5-azido-3-(tert-butyldimethylsilyloxy)-1-hydroxy-4-methylpenta-
n-2-ylcarbamate
##STR00101##
[0322] To a solution of (R)-tert-butyl
4-((1R,2S)-3-azido-1-(tert-butyldimethylsilyloxy)-2-methylpropyl)-2,2-dim-
ethyloxazolidine-3-carboxylate (1.0 equiv.) in EtOH (0.1 M) was
added PPTS (1.3 equiv.) and the mixture was refluxed for 2 days.
The volatiles were removed under vacuo, the residue was dissolved
in DCM (0.1 M) and DIEA (1.5 equiv.) and Boc.sub.2O (1.0 equiv.)
were added to the reaction mixture. The solution was stirred for 3
hours at room temperature. The solvents were removed under reduced
pressure and the residue was diluted with ethyl acetate, washed
with water, aqueous NaHSO.sub.4, aqueous NaHCO.sub.3, sat. NaCl,
the organic phase was dried with magnesium sulfate, filtered, and
concentrated. The residue was purified via silica gel column
chromatography eluting with ethyl acetate and hexanes (1:3) to give
tert-butyl
(2R,3R,4S)-5-azido-3-(tert-butyldimethylsilyloxy)-1-hydroxy-4-methylpenta-
n-2-ylcarbamate as the major isomer (5:2) in 70% yield. LC/MS=289.3
(M+H-Boc), Rt=0.76 min (Frac 65%-95% method).
Synthesis of
(2R,3R,4S)-5-azido-2-(tert-butoxycarbonylamino)-3-(tert-butyldimethylsily-
loxy)-4-methylpentyl methanesulfonate
##STR00102##
[0324] To a solution of tert-butyl
(2R,3R,4S)-5-azido-3-(tert-butyldimethylsilyloxy)-1-hydroxy-4-methylpenta-
n-2-ylcarbamate (1.0 equiv.) in pyridine (0.2 M) was added MsCl
(1.3 equiv.) followed by DMAP (catalytic amount) at 0.degree. C.
The mixture was stirred at that temperature for 1 hour. The
solution was diluted with ether and ethyl acetate (4:1), washed
with aq. NaHSO.sub.4, sat. NaHCO.sub.3, brine, dried over magnesium
sulfate, filtered, and concentrated. The residue was purified by
silica gel column chromatography eluting with ethyl acetate and
hexanes (1:3) to give
(2R,3R,4S)-5-azido-2-(tert-butoxycarbonylamino)-3-(tert-butyldimethylsily-
loxy)-4-methylpentyl methanesulfonate as the major isomer (5:2) in
90% yield. LC/MS=367.3 (M+H-Boc), Rt=0.81 min (Frac 65%-95%
method).
Synthesis of tert-butyl
(3R,4R,5S)-4-(tert-butyldimethylsilyloxy)-5-methylpiperidin-3-ylcarbamate
##STR00103##
[0326] A solution of
(2R,3R,4S)-5-azido-2-(tert-butoxycarbonylamino)-3-(tert-butyldimethylsily-
loxy)-4-methylpentyl methanesulfonate in MeOH (0.09 M) was degassed
with nitrogen for 20 min. DIEA (2.5 equiv.) was added, followed by
10% Pd/C (0.1 equiv.). The reaction mixture was stirred under a
hydrogen balloon for 2 hours. The solution was filtered and the
filtrate was concentrated under vacuo to afford tert-butyl
(3R,4R,5S)-4-(tert-butyldimethylsilyloxy)-5-methylpiperidin-3-ylcarbamate
as the major isomer (5:2) in >99% yield. LC/MS=345.2 (M+H-Boc),
Rt=0.95 and 0.99 min.
Synthesis of tert-butyl
(3R,4R,5S)-4-(tert-butyldimethylsilyloxy)-5-methyl-1-(3-nitropyridin-4-yl-
)piperidin-3-ylcarbamate
##STR00104##
[0328] To a solution of tert-butyl
(3R,4R,5S)-4-(tert-butyldimethylsilyloxy)-5-methylpiperidin-3-ylcarbamate
(1.0 equiv.) in i-PrOH (0.09 M) was added DIEA (2.5 equiv.) and
4-chloro-3-nitropyridine (1.5 equiv.). The reaction mixture was
stirred at 60.degree. C. for 2 hours. The volatiles were removed
under vacuo, the residue was diluted with ethyl acetate and washed
with sat. NaCl. The organic phase was dried with magnesium sulfate,
filtered, and concentrated. The crude material was purified by
silica gel column chromatography eluting with ethyl acetate and
hexanes (1:2) to give tert-butyl
(3R,4R,5S)-4-(tert-butyldimethylsilyloxy)-5-methyl-1-(3-nitropyridin-4-yl-
)piperidin-3-ylcarbamate in 76% yield. LC/MS=467.3 (M+H), Rt=1.09
min.
Synthesis of tert-butyl
(3R,4R,5S)-1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)-5-methy-
lpiperidin-3-ylcarbamate
##STR00105##
[0330] A solution of tert-butyl
(3R,4R,5S)-4-(tert-butyldimethylsilyloxy)-5-methyl-1-(3-nitropyridin-4-yl-
)piperidin-3-ylcarbamate (1.0 equiv.) in MeOH (0.05 M) was degassed
with nitrogen for 20 min. 10% Pd/C (0.2 equiv.) was added to the
mixture and the solution was stirred under a hydrogen balloon for 3
hours. The reaction was filtered and the filtrate was concentrated
under reduced pressure to give tert-butyl
(3R,4R,5S)-1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)-5-methy-
lpiperidin-3-ylcarbamate as the desired product in 94% yield.
LC/MS=437.4 (M+H), Rt=1.08 min. .sup.1H-NMR (300 MHz, CDCl.sub.3):
.delta. 8.01 (s, 1H), 7.95 (d, J=6.0 Hz, 1H), 6.76 (d, J=6.0 Hz,
1H), 4.44 (br s, 1H), 3.74 (br s, 2H), 3.59-3.55 (m, 1H), 3.25-3.13
(m, 2H), 2.47-2.35 (m, 2H), 1.89 (br s, 2H), 1.44 (s, 9H), 1.04 (d,
J=6.0, 3H), 0.92 (s, 9H), 0.13 (d, J=9.0, 6H).
Synthesis of
5-methyl-3-oxocyclohex-1-enyltrifluoromethanesulfonate
##STR00106##
[0332] To a solution of 5-methylcyclohexane-1,3-dione (1.0 equiv.)
in DCM (0.5M) was added Na.sub.2CO.sub.3 (1.1 equiv.) and cooled to
0.degree. C. Added Tf.sub.2O (1.0 equiv.) in DCM (5.0 M) dropwise
over 1 hr at 0.degree. C. under a nitrogen atmosphere. Upon
addition, the reaction was stirred for 1 hr at room temperature
(dark red solution). The solution was filtered and the filtrate was
quenched by careful addition of saturated NaHCO.sub.3 with vigorous
stirring until pH=7. The solution was transferred to a separatory
funnel and the layers were separated. The organic layer was washed
with brine, dried with Na.sub.2SO.sub.4, filtered, concentrated
under vacuo and dried under high vacuum for 15 min to yield
5-methyl-3-oxocyclohex-1-enyl trifluoromethanesulfonate as light
yellow oil in 78% yield. The triflate decomposes upon storage and
should be used immediately for the next reaction. LC/MS=259.1/300.1
(M+H and M+CH.sub.3CN); Rt=0.86 min, LC=3.84 min. .sup.1H-NMR (400
MHz, CDCl.sub.3) .delta. ppm: 6.05 (s, 1H), 2.70 (dd, J=17.2, 4.3,
1H), 2.53 (dd, J=16.6, 3.7, 1H), 2.48-2.31 (m, 2H), 2.16 (dd,
J=16.4, 11.7, 1H), 1.16 (d, J=5.9, 3H).
Synthesis of
5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-2-enone
##STR00107##
[0334] To a solution of 5-methyl-3-oxocyclohex-1-enyl
trifluoromethanesulfonate (1.0 equiv.) in degassed dioxane (0.7 M)
was added bis(pinacolato)diboron (2.0 equiv.), KOAc (3.0 equiv.),
and Pd(dppf)Cl.sub.2-DCM (0.03 equiv.). The reaction was heated to
80.degree. C. for 10 h then cooled to room temperature and filtered
through a coarse frit glass funnel. The cake was rinsed with more
dioxane and the filtrate solution was used for the next step
without further purification. LC/MS=155.1 (M+H of boronic acid);
Rt=0.41 min, LC=1.37 min.
Synthesis of 5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enone
##STR00108##
[0336] To a solution of
5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-2-enone
(1.0 equiv.) in degassed dioxane (0.5 M) and 2M Na.sub.2CO.sub.3 (2
equiv.) was added 4-chloro-3-nitropyridine (1.3 equiv.) and
Pd(dppf)Cl.sub.2-DCM (0.05 equiv.). The reaction was placed under a
reflux condenser and heated in an oil bath to 110.degree. C. for 1
h. Cooled to room temperature, filtered through a pad of Celite,
washed the pad with ethyl acetate and concentrated the filtrate
under vacuo. The residue was further pumped at 80.degree. C. on a
rotary evaporator for one hour to remove boronate by-products
(M+H=101) via sublimation. The residue was partitioned between
brine and ethyl acetate, and the layers were separated, the aqueous
phase was further extracted with ethyl acetate (4.times.), the
organics were combined, dried over sodium sulfate, filtered, and
concentrated. The crude was purified via silica gel chromatography
loading in DCM and eluting with 2-50% ethyl acetate and hexanes.
The pure fractions were concentrated in vacuo to yield an orange
oil. The oil was placed under high vacuum (.about.500 mtorr) with
seed crystals overnight to yield an orange solid. The solid was
further purified via trituration in hexanes to yield
5-methyl-3-(3-nitropyridin-4-yl) cyclohex-2-enone (48% 2 steps).
LC/MS=233.2 (M+H); Rt=0.69 min, LC=2.70 min. .sup.1H-NMR (400 MHz,
CdCl.sub.3) .delta. ppm: 9.31 (s, 1H), 8.88 (d, J=5.1, 1H), 7.30
(d, J=5.1, 1H), 6.00 (d, J=2.4, 1H), 2.62 (dd, J=16.4, 3.5, 1H),
2.53-2.34 (m, 3H), 2.23 (dd, J=16.1, 11.7, 1H), 1.16 (d, J=6.3,
3H).
Synthesis of
cis-(+/-)-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enol
##STR00109##
[0338] To a solution of
5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enone (1.0 equiv.) in
EtOH (0.3 M) was added CeCl.sub.3-7H.sub.2O (1.2 equiv.). The
reaction was cooled to 0.degree. C., then NaBH.sub.4 (1.2 equiv.)
was added in portions. Stirred for 1 h at 0.degree. C., then
quenched by adding water, concentrated to remove the EtOH, added
EtOAc, extracted the organics, washed with brine, then dried with
Na.sub.2SO.sub.4, filtered and concentrated to yield
cis-(+/-)-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enol (94%).
LC/MS=235.2 (M+H), LC=2.62 min.
Synthesis of
(+/-)-4-(5-methylcyclohexa-1,3-dienyl)-3-nitropyridine
##STR00110##
[0340] To a solution of
(+/-)-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enol (1.0 equiv.)
in dioxane (0.1M) was added p-TSA (1.0 equiv.), and the reaction
was stirred at 100.degree. C. for 3 h. The solution was cooled to
room temperature, then passed through a pad of neutral alumina
eluting with EtOAc to yield
(+/-)-4-(5-methylcyclohexa-1,3-dienyl)-3-nitropyridine as a yellow
oil in 68% yield. LC/MS=217.1 (M+H), LC=3.908 min.
Synthesis of
(+/-)-6-bromo-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enol
##STR00111##
[0342] To a solution of
4-(5-methylcyclohexa-1,3-dienyl)-3-nitropyridine (1.0 equiv.) in
THF and water (1:1, 0.13 M) was added NBS (1.5 equiv.) and the
reaction was stirred at room temperature for 30 min. Upon
completion, ethyl acetate and water were added to the reaction, the
organic phase was dried with brine, then sodium sulfate, filtered,
and concentrated. The crude material was purified via silica gel
column chromatography eluting with ethyl acetate and hexanes (1:1)
to give
(+/-)-6-bromo-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enol as a
yellow oil in 80% yield. LC/MS=315.0/313.0 (M+H), LC=2.966 min.
Synthesis of
(+/-)-2-azido-6-methyl-4-(3-nitropyridin-4-yl)cyclohex-3-enol
##STR00112##
[0344] To a solution of
(+/-)-6-bromo-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enol (1.0
equiv.) in THF (0.1 M) was added potassium tert-butoxide (1.5
equiv.). The reaction turned from orange to black almost
immediately. By TLC, the formation of product is clean in 30 min.
Quenched by adding saturated ammonium chloride and ethyl acetate.
The organic phase was dried with brine, then sodium sulfate,
filtered, and concentrated. The crude product was dissolved in
ethanol and water (3:1, 0.1 M), and ammonium chloride (2.0 equiv)
and sodium azide (2.0 equiv.) were added. The dark orange reaction
was stirred at room temperature overnight. The conversion to
product is clean as indicated by LC/MS. The reaction was
concentrated to remove the ethanol, ethyl acetate and water were
added, and the organic phase was dried with sodium sulfate,
filtered, and concentrated. The crude material was purified via
silica gel column chromatography eluting with ethyl acetate and
hexanes (1:1) to give
(+/-)-2-azido-6-methyl-4-(3-nitropyridin-4-yl)cyclohex-3-enol in
55% yield. LC/MS=276.0 (M+H), LC=2.803 min.
Synthesis of (+/-)-tert-butyl
6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enylcarbamate
##STR00113##
[0346] To a solution of
(+/-)-2-azido-6-methyl-4-(3-nitropyridin-4-yl)cyclohex-3-enol (1.0
equiv.) in pyridine and ammonium hydroxide (8:1, 0.08 M) was added
trimethylphosphine (3.0 equiv.) and the brown solution was stirred
at room temperature for 2 h. Upon completion, EtOH was added and
the solution was concentrated in vacuo. More ethanol was added and
the reaction was concentrated again. Dioxane and sat. NaHCO.sub.3
(1:1, 0.08 M) were added to the crude, followed by Boc.sub.2O (1.0
equiv.). Stirred the reaction mixture at room temperature for 2 h,
then added water and ethyl acetate. The organic phase was dried
with MgSO.sub.4, and concentrated. The crude product was purified
via silica gel column chromatography eluting with ethyl acetate and
hexanes (1:1) to afford (+/-)-tert-butyl
6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enylcarbamate
(59%). LC/MS=350.1 (M+H), Rt: 0.76 min.
Synthesis of
(+/-)-2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-yl)cycloh-
ex-3-enyl acetate
##STR00114##
[0348] To a solution of (+/-)-tert-butyl
6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enylcarbamate
(1.0 equiv.) in pyridine (0.1 M) was added Ac.sub.2O (2.0 equiv.)
and the reaction was stirred at room temperature overnight. Upon
completion, the reaction was concentrated to dryness, then
worked-up with ethyl acetate and water. The organic phase was dried
with brine, then sodium sulfate, filtered, and concentrated to give
(+/-)-2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-yl)cycloh-
ex-3-enyl acetate in 94% yield. LC/MS=392.2 (M+H), Rt=0.94 min.
Synthesis of (1
S,2S,4S,6R)-4-(3-aminopyridin-4-yl)-2-((tert-butoxycarbonyl)amino)-6-meth-
ylcyclohexyl acetate and
(1R,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-((tert-butoxycarbonyl)amino)-6-me-
thylcyclohexyl acetate
##STR00115##
[0350] To a degassed solution of
(+/-)-2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-yl)cycloh-
ex-3-enyl acetate (1.0 equiv.) in MeOH and EtOAc (1:1, 0.1 M) was
added 10% Pd/C (0.1 equiv.) and the reaction was stirred at room
temperature under a hydrogen balloon for 3 days. Upon completion,
the solution was filtered through a pad of Celite, the pad was
washed with ethyl acetate and the filtrate was concentrated. The
crude material contained about 10% of the undesired isomer. The
crude was dissolved in ethyl acetate (.about.20%) and hexanes and
heated until all dissolved. The solution was allowed to sit at room
temperature for 2 days. The precipitate was then collected to give
(+/-)-4-(3-aminopyridin-4-yl)-2-(tert-butoxycarbonylamino)-6-methylcycloh-
exyl acetate as the pure product in 59% yield. LC/MS=364.3 (M+H),
Rt=0.63 min. The racemic material was resolved using an AD-H chiral
column (20% i-PrOH/80% n-heptanes, 20 mL/min flow rate) to
(1S,2S,4S,6R)-4-(3-aminopyridin-4-yl)-2-((tert-butoxycarbonyl)amino)-6-me-
thylcyclohexyl acetate (peak#1, R.sub.t=3.76 min on AD-H chiral
analytical column, 20% i-PrOH/80% n-heptanes, 1 mL/min) and
(1R,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-((tert-butoxycarbonyl)amino)-6-me-
thylcyclohexyl acetate (peak#2, R.sub.t =6.79 min on AD-H chiral
analytical column, 20% i-PrOH/80% n-heptanes, 1 mL/min).
Synthesis of
2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-yl)cyclohex-3-e-
nyl methanesulfonate
##STR00116##
[0352] To a solution of tert-butyl
6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enylcarbamate
(1.0 equiv.) in DCM (0.09 M) was added triethylamine (1.5 equiv.)
and the reaction was cooled to 0.degree. C. MsCl (1.2 equiv.) was
added to the reaction and stirred for 3 h. Another 1.0 equiv. of
MsCl was added to the reaction and stirred for another 2 h. Worked
up the reaction by adding water, the organic phase was dried with
brine, sodium sulfate, and concentrated. The crude product was
purified via silica gel column chromatography eluting with ethyl
acetate and hexanes (1:1) to afford
2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-yl)cyclohex-3-e-
nyl methanesulfonate as a white foam in 65% yield. LC/MS=428.2
(M+H), LC: 3.542 min.
Synthesis of (+/-)-tert-butyl
7-methyl-5-(3-nitropyridin-4-yl)-2-oxo-3a,6,7,7a-tetrahydrobenzo[d]oxazol-
e-3(2H)-carboxylate
##STR00117##
[0354] A solution of
(+/-)-2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-yl)cycloh-
ex-3-enyl methanesulfonate (1.0 equiv.) in pyridine (0.2 M) was
heated in the microwave at 110.degree. C. for 10 min. The orange
reaction was then concentrated under vacuo, the crude was dissolved
in ethyl acetate and water, the organic phase was dried with sodium
sulfate and concentrated under vacuo. The crude material was
dissolved in DCM (0.2 M), triethylamine (1.8 equiv.) was added,
followed by Boc.sub.2O (1.2 equiv.). The reaction was stirred for
40 min, then concentrated to dryness. The crude material was
purified via silica gel column chromatography eluting with hexane
and ethyl acetate (1:1) to afford (+/-)-tert-butyl
7-methyl-5-(3-nitropyridin-4-yl)-2-oxo-3a,6,7,7a-tetrahydrobenzo[d]oxazol-
e-3(2H)-carboxylate as a white foam in 66% yield. LC/MS=376.0
(M+H), LC: 3.424 min.
Synthesis of (+/-)-tert-butyl
5-(3-aminopyridin-4-yl)-7-methyl-2-oxohexahydrobenzo[d]oxazole-3(2H)-carb-
oxylate
##STR00118##
[0356] To a degassed solution of (+/-)-tert-butyl
7-methyl-5-(3-nitropyridin-4-yl)-2-oxo-3a,6,7,7a-tetrahydrobenzo[d]oxazol-
e-3(2H)-carboxylate (1.0 equiv.) in MeOH and EtOAc (1:1, 0.1 M) was
added 10% Pd/C (0.1 equiv.). The reaction was stirred under a
hydrogen balloon overnight. Upon completion, the solution was
filtered through a pad of Celite and the pad was washed with ethyl
acetate. The filtrate was concentrated under vacuo to give
(+/-)-tert-butyl
5-(3-aminopyridin-4-yl)-7-methyl-2-oxohexahydrobenzo[d]oxazole-3(2H)-carb-
oxylate as the desired product as a yellow foam in 93% yield.
LC/MS=348.1 (M+H), Rt=055 min.
Synthesis of tert-butyl
(1R,2R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-2-hydroxy-3-methylcyclohexylcarbamate
##STR00119##
[0358] To a solution of
(1R,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-(tert-butoxycarbonylamino)-6-meth-
ylcyclohexyl acetate (1.0 equiv.) in DMF/Ethanol (1/5, 0.05 M) was
added 6-(2,6-difluorophenyl)-5-fluoropicolinic acid (1.3 equiv.),
aza-HOBt (1.3 equiv.) and EDC (1.3 equiv.). The mixture was stirred
at rt for 6 hrs. The solution was diluted with EtOAc, washed with
H.sub.2O, 1N NaOH, NaCl (sat.), dried over MgSO4, filtered and
concentrated to yield crude protected amide. The material was
dissolved in EtOH (0.45 M), Cs.sub.2CO.sub.3 (1.0 equiv.) was added
and the solution was submerged in a 60.degree. C. oil bath and
stirred for 90 mins. The volatiles were removed in vacuo; the
residue was partitioned between with EtOAc and H.sub.2O. The
organic layer was washed with NaCl.sub.(sat.), dried over
MgSO.sub.4, filtered, concentrated and pumped on to yield
tert-butyl
(1R,2R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-2-hydroxy-3-methylcyclohexylcarbamate in 99% yield. LC/MS
(m/z)=557.3 (MH.sup.+), R.sub.t=0.80 min.
Synthesis of
(1R,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexyl methanesulfonate
##STR00120##
[0360] To a solution of tert-butyl
(1R,2R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-2-hydroxy-3-methylcyclohexylcarbamate (1.0 equiv.) in
CH.sub.2Cl.sub.2 was added TEA (4.0 equiv.) and MsCl (2.0 equiv.).
The capped solution was stirred for 5 minutes and then the
homogeneous solution was left standing at rt for 16 hrs. The
volatiles were removed in vacuo, the residue was dissolved in DMSO,
purified by RP HPLC and the product fractions were lyophilized
directly. The Boc protected product was treated with 25%
TFA/CH.sub.2Cl.sub.2 for 20 minutes at which time the volatiles
were removed in vacuo and the residue was dissolved in DMSO and
purified by RP-HPLC. The product fractions were lyophilized
directly to yield
(1R,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicol-
inamido)pyridin-4-yl)-6-methylcyclohexyl methanesulfonate in 31%
yield. LC/MS (m/z)=535.2 (MH.sup.+), R.sub.t=0.61 min.
Synthesis of tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-2-hydroxy-3-methylcyclohexylcarbamate
##STR00121##
[0362] To a solution of (3aR,5R,7S,7aS)-tert-butyl
5-(3-aminopyridin-4-yl)-7-methyl-2-oxohexahydrobenzo[d]oxazole-3(2H)-carb-
oxylate (1.0 equiv.) in DMF (0.2 M) was added
6-(2,6-difluorophenyl)-5-fluoropicolinic acid (1.3 equiv.),
aza-HOBt (1.3 equiv.) and EDC (1.3 equiv.). The mixture was stirred
at rt for 16 hrs. The solution was diluted with EtOAc, washed with
H.sub.2O, 1N NaOH, NaCl.sub.(sat.), dried over MgSO.sub.4, filtered
and concentrated to yield crude protected amide. The material was
dissolved in EtOH (0.45 M), Cs.sub.2CO.sub.3 (1.0 equiv.) was added
and the solution was submerged in a 60.degree. C. oil bath and
stirred for 90 mins. The volatiles were removed in vacuo; the
residue was partitioned between with EtOAc and H.sub.2O. The
organic layer was washed with NaCl.sub.(sat.), dried over
MgSO.sub.4, filtered, concentrated and pumped on to yield
tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-2-hydroxy-3-methylcyclohexylcarbamate in 100% yield. LC/MS
(m/z)=557.3 (MH.sup.+), R.sub.t=0.83 min.
Synthesis of
(1S,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexyl methanesulfonate
##STR00122##
[0364] To a solution of tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-2-hydroxy-3-methylcyclohexylcarbamate (1.0 equiv.) in pyridine
(0.12 M) was added MsCl (7.0 equiv.). The capped solution was
stirred for 5 minutes and then the homogeneous solution was left
standing at rt for 16 hrs. The volatiles were removed in vacuo and
the residue was partitioned between EtOAc and H.sub.2O. The organic
layer was washed with H.sub.2O, 10% CuSO.sub.4, H.sub.2O,
Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.), dried over MgSO.sub.4,
filtered, concentrated and purified by ISCO chromatography. The Boc
protected product was treated with 25% TFA/CH.sub.2Cl.sub.2 for 20
minutes at which time the volatiles were removed in vacuo and the
residue was dissolved in DMSO and purified by RP-HPLC. The product
fractions were lyophilized directly to yield
(1S,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexyl methanesulfonate in 45% yield.
LC/MS (m/z)=535.2 (MH.sup.+), R.sub.t=0.60 min.
Synthesis of
(1S,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexyl dimethylphosphinate
##STR00123##
[0366] To a solution of tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-2-hydroxy-3-methylcyclohexylcarbamate (1.0 equiv.) in pyridine
(0.05 M) was added phosphonic chloride (5.0 equiv.). The capped
homogeneous solution was left stirring at rt for 1 hr. The
volatiles were removed in vacuo and the residue was partitioned
between EtOAc and H.sub.2O. The organic layer was washed with
H.sub.2O, 10% CuSO.sub.4, Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.),
dried over MgSO.sub.4, filtered, concentrated to yield crude Boc
protected product. The Boc group was removed by treating with 25%
TFA/CH.sub.2Cl.sub.2 for 30 minutes at which time the volatiles
were removed in vacuo and the residue was dissolved in DMSO and
purified by RP-HPLC. The product fractions were lyophilized
directly to yield (1
S,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyr-
idin-4-yl)-6-methylcyclohexyl dimethylphosphinate in 43% yield.
LC/MS (m/z)=533.3 (MH.sup.+), R.sub.t=0.59 min.
Synthesis of
(1R,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexyl dimethylphosphinate
##STR00124##
[0368] To a solution of tert-butyl
(1R,2R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-2-hydroxy-3-methylcyclohexylcarbamate (1.0 equiv.) in pyridine
(0.05 M) was added phosphonic chloride (5.0 equiv.). The capped
homogeneous solution was left stirring at rt for 1 hr. The
volatiles were removed in vacuo and the residue was partitioned
between EtOAc and H.sub.2O. The organic layer was washed with
H.sub.2O, 10% CuSO.sub.4, Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.),
dried over MgSO.sub.4, filtered, concentrated to yield crude Boc
protected product. The Boc group was removed by treating with 25%
TFA/CH.sub.2Cl.sub.2 for 30 minutes at which time the volatiles
were removed in vacuo and the residue was dissolved in DMSO and
purified by RP-HPLC. The product fractions were lyophilized
directly to yield
(1R,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexyl dimethylphosphinate in 43% yield.
LC/MS (m/z)=533.3 (MH.sup.+), R.sub.t=0.60 min.
Synthesis of
S-(1S,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl)-
-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl
ethanethioate
##STR00125##
[0370] To a solution of
(1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl)-5-
-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl
methanesulfonate (1.0 equiv.) in DMF (0.19 M) was added KSAc (3.0
equiv.). The capped solution was left stirring at rt for 20 hrs.
The solution was partitioned between EtOAc and H.sub.2O. The
organic layer was washed with H.sub.2O, Na.sub.2CO.sub.3(sat.),
NaCl.sub.(sat.), dried over MgSO.sub.4, filtered, concentrated and
purified by ISCO SiO.sub.2 chromatography to yield
S-(1S,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl)-
-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl
ethanethioate in 35% yield. LC/MS (m/z)=615.2 (MH.sup.+),
R.sub.t=0.95 min.
Synthesis of tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylthio)cyclohexylcarbamate
##STR00126##
[0372] To a solution of
S-(1S,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl)-
-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl
ethanethioate (1.0 equiv.) in MeOH (0.07 M) was added
K.sub.2CO.sub.3 (3.0 equiv.). The heterogeneous solution was capped
and left stirring at rt for 1 hr. Methyl iodide (1.5 equiv.) was
added and stirred at rt for 10 min. The volatiles were removed in
vacuo and the residue was partitioned between EtOAc and H.sub.2O.
The organic layer was washed with NaCl.sub.(sat.), dried over
MgSO.sub.4, filtered, concentrated and purified by ISCO SiO.sub.2
chromatography to yield tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)
pyridin-4-yl)-3-methyl-2-(methylthio)cyclohexylcarbamate in 78%
yield. LC/MS (m/z)=587.2 (MH.sup.+), R.sub.t=1.00 min.
Synthesis of
N-(4-((1R,3R,4S,5S)-3-amino-5-methyl-4-(methylthio)cyclohexyl)pyridin-3-y-
l)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00127##
[0374] To a solution of HCl (30.0 equiv.) in dioxane was added to
tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylthio)cyclohexylcarbamate (1.0 equiv.). The
solution was capped and left standing at rt for 1 hr. The volatiles
were removed in vacuo, dissolved in DMSO and purified by reverse
phase HPLC to yield
N-(4-((1R,3R,4S,5S)-3-amino-5-methyl-4-(methylthio)cyclohexyl)pyridin-3-y-
l)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 94% yield. LC/MS
(m/z)=487.2 (MH.sup.+), R.sub.t=0.65 min. .sup.1H NMR (400 MHz,
<dmso>) .delta. ppm 10.44 (s, 1H), 8.58 (d, J=4.0, 1H), 8.47
(d, J=4.0, 1H), 8.34 (dd, J=8.0, 4.0, 1H), 8.20 (dd, J=8.0, 8.0,
1H), 8.20 (dd, J=16.0, 4.0, 1H), 7.67-7.74 (m, 1H), 7.36 (dd,
J=8.0, 8.0, 2H), 7.26 (d, J=4.0, 1H), 2.85-2.95 (m, 2H), 2.18 (s,
3H), 1.88-1.98 (m, 1H), 1.74-1.84 (m, 1H), 1.48-1.56 (m, 1H),
1.38-1.48 (m, 1H), 1.18-1.28 (m, 1H), 1.02 (d, J=8.0, 3H).
Synthesis of tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl)-2-((R)-methylsulfinyl)cyclohexylcarbamate and
tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylsulfonyl)cyclohexylcarbamate
##STR00128##
[0376] To a solution of tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylthio)cyclohexylcarbamate (1.0 equiv.) in
CH.sub.2Cl.sub.2 (0.10 M) in a 0.degree. C. bath was added mCPBA
(1.2 equiv.). The capped solution was left stirring at rt for 1 hr.
cyclohexene (10.0 equiv.) was added to quench any remaining mCBPA
and after stirring for 5 minutes the solution was directly loaded
onto ISCO SiO.sub.2 column and purified to yield tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-((R)-methylsulfinyl)cyclohexylcarbamate in 31%
yield, LC/MS=603.2 (MH+), Rt=0.66 min; and tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylsulfonyl)cyclohexylcarbamate in 37% yield.
LC/MS (m/z)=619.2 (MH.sup.+), R.sub.t=0.88 min.
Synthesis of
N-(4-((1R,3R,4S,5S)-3-amino-5-methyl-4-((R)-methylsulfinyl)cyclohexyl)pyr-
idin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00129##
[0378] To a solution of tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-((R)-methylsulfinyl)cyclohexylcarbamate in DCM
(0.10 M) was added TFA (30.0 equiv.). The solution was capped and
left standing at rt for 1 hr. The volatiles were removed in vacuo;
the residue was dissolved in DMSO and purified by reverse phase
HPLC. The product fractions were lyophilized directly to yield
N-(4-((1R,3R,4S,5S)-3-amino-5-methyl-4-((R)-methylsulfinyl)cyclohexyl)pyr-
idin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 54%
yield. LC/MS (m/z)=503.2 (MH.sup.+), Rt=0.57 min. 1H NMR (400 MHz,
<dmso>) .delta. ppm 10.48 (s, 1H), 8.58 (d, J=4.0, 1H), 8.50
(d, J=4.0, 1H), 8.34 (dd, J=8.0, 4.0, 1H), 8.20 (dd, J=8.0, 8.0,
1H), 8.20 (dd, J=16.0, 4.0, 1H), 7.67-7.74 (m, 1H), 7.42 (d, J=4.0,
1H), 7.36 (dd, J=8.0, 8.0, 2H), 3.40-3.42 (m, 1H), 3.06-3.20 (m,
1H), 2.92 (s, 3H), 2.06-2.20 (m, 1H), 1.95-2.04 (m, 2H), 1.70-1.80
(m, 1H), 1.56-1.70 (m, 1H), 0.86 (d, J=8.0, 3H).
Synthesis of
N-(4-((1R,3R,4S,5S)-3-amino-5-methyl-4-(methylsulfonyl)cyclohexyl)pyridin-
-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00130##
[0380] To a solution of tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylsulfonyl)cyclohexylcarbamate in DCM (0.02 M)
was added TFA (30.0 equiv.). The solution was capped and left
standing at rt for 1 hr. The volatiles were removed in vacuo; the
residue was dissolved in DMSO and purified by reverse phase HPLC.
The product fractions were lyophilized directly to yield
N-(4-((1R,3R,4S,5S)-3-amino-5-methyl-4-(methylsulfonyl)cyclohexyl)pyridin-
-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 73% yield.
LC/MS (m/z)=519.2 (MH.sup.+), R.sub.t=0.58 min. 1H NMR (400 MHz,
<dmso>) .delta. ppm 10.52 (s, 1H), 8.51-8.54 (m, 2H), 8.34
(dd, J=8.0, 4.0, 1H), 8.20 (dd, J=8.0, 8.0, 1H), 8.15 (dd, J=16.0,
4.0, 1H), 7.67-7.74 (m, 1H), 7.36 (dd, J=8.0, 8.0, 2H), 7.28 (d,
J=4.0, 1H), 3.77-3.79 (m, 1H), 3.18 (s, 3H), 3.02-3.20 (m, 1H),
1.94-2.40 (m, 5H), 1.57-1.62 (m, 1H), 1.24 (d, J=8.0, 3H).
Synthesis of tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-2-(2-methoxyethylthio)-3-methylcyclohexylcarbamate
##STR00131##
[0382] To a solution of
S-(1S,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl)-
-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl
ethanethioate (1.0 equiv.) in MeOH (0.04 M) was added
K.sub.2CO.sub.3 (3.0 equiv.). The heterogeneous solution was capped
and left stirring at rt for 1 hr. 1-bromo-2-methoxyethane (7.0
equiv.) was added and stirred at rt for 6 hrs. Quench the reaction
with diisopropylamine and the volatiles were removed in vacuo and
the residue was partitioned between EtOAc and H.sub.2O. The organic
layer was washed with NaCl.sub.(sat.), dried over MgSO.sub.4,
filtered, concentrated and purified by ISCO SiO.sub.2
chromatography to yield tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-2-(2-methoxyethylthio)-3-methylcyclohexylcarbamate in 63%
yield. LC/MS (m/z)=631.2 (MH.sup.+), R.sub.t=0.98 min.
Synthesis of
N-(4-((1R,3R,4S,5S)-3-amino-4-(2-methoxyethyl)sulfonyl)-5-methylcyclohexy-
l)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00132##
[0384] To a solution of tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-2-(2-methoxyethylthio)-3-methylcyclohexylcarbamate (1.0
equiv.) in THF (0.04 M) in a 0.degree. C. bath was added oxone (2.0
equiv.) as a solution in H.sub.2O. The solution was left stirring
at rt for 5 hrs. The solution was diluted with EtOAc, washed with
H.sub.2O, NaCl.sub.(sat.), dried over MgSO.sub.4, filtered,
concentrated to yield Boc protected product. The material was
treated with 25% TFA/CH.sub.2Cl.sub.2 for 30 minutes, at which time
the volatiles were removed in vacuo, the residue was dissolved in
DMSO and purified by reverse phase HPLC. The product fractions were
lyophilized directly to yield
N-(4-((1R,3R,4S,5S)-3-amino-4-((2-methoxyethyl)sulfonyl)-5-methylcyclohex-
yl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 76%
yield. LC/MS (m/z)=563.3 (MH.sup.+), R.sub.t=0.62 min. .sup.1H NMR
(400 MHz, <dmso>) .delta. ppm 10.52 (s, 1H), 8.53 (s, 1H),
8.50 (d, J=4.0, 1H), 8.35 (dd, J=8.0, 4.0, 1H), 8.20 (t, J=8.0,
1H), 7.80 (bs, 2H), 7.70 (quintet, J=8.0, 1H), 7.36 (t, J=8.0, 2H),
7.31 (d, J=4.0, 1H), 3.82-3.88 (m, 2H), 3.73-3.78 (m, 2H),
3.55-3.61 (m, 1H), 3.42-3.49 (m, 1H), 3.31 (s, 3H), 3.06-3.16 (m,
1H), 1.92-2.18 (m, 4H), 1.54-1.64 (m, 1H), 1.22 (d, J=4.0, 3H).
Synthesis of tert-butyl
(1R,2S,3S,5R)-2-(2-(tert-butyldimethylsilyloxy)ethylthio)-5-(3-(6-(2,6-di-
fluorophenyl)-5-fluoropicolinamido)pyridin-4-yl)-3-methylcyclohexylcarbama-
te
##STR00133##
[0386] To a solution of
S-(1S,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl)-
-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl
ethanethioate (1.0 equiv.) in MeOH (0.04 M) was added
K.sub.2CO.sub.3 (5.0 equiv.). The heterogeneous solution was capped
and left stirring at rt for 1 hr.
(2-bromoethoxy)(tert-butyl)dimethylsilane (7.0 equiv.) was added
and stirred at rt for 6 hrs. Quench the reaction with
diisopropylamine and the volatiles were removed in vacuo and the
residue was partitioned between EtOAc and H.sub.2O. The organic
layer was washed with NaCl.sub.(sat.), dried over MgSO.sub.4,
filtered, concentrated. The residue was dissolved in DMF (0.05 M)
and imidazole (20.0 equiv.) and TBDMSCl (7.0 equiv.) were added.
After stirring at rt for 1 hr, the solution was partitioned between
EtOAc and H.sub.2O. The organic layer was washed with
NaCl.sub.(sat.), dried over MgSO.sub.4, filtered, concentrated and
purified by ISCO SiO.sub.2 chromatography to yield tert-butyl
(1R,2S,3S,5R)-2-(2-(tert-butyldimethylsilyloxy)ethylthio)-5-(3-(6-(2,6-di-
fluorophenyl)-5-fluoropicolinamido)pyridin-4-yl)-3-methylcyclohexylcarbama-
te in 57% yield. LC/MS (m/z)=731.4 (MH.sup.+), R.sub.t=1.27
min.
Synthesis of
N-(4-((1R,3R,4S,5S)-3-amino-4-(2-hydroxyethylsulfonyl)-5-methylcyclohexyl-
)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00134##
[0388] To a solution of tert-butyl
(1R,2S,3S,5R)-2-(2-(tert-butyldimethylsilyloxy)ethylthio)-5-(3-(6-(2,6-di-
fluorophenyl)-5-fluoropicolinamido)pyridin-4-yl)-3-methylcyclohexylcarbama-
te (1.0 equiv.) in THF (0.04 M) in a 0.degree. C. bath was added
oxone (2.0 equiv.) as a solution in H.sub.2O. The solution was left
stirring at rt for 48 hrs. The solution was diluted with EtOAc,
washed with H.sub.2O, Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.),
dried over MgSO.sub.4, filtered, concentrated to yield Boc
protected product. The material was treated with 25%
TFA/CH.sub.2Cl.sub.2 for 30 minutes, at which time the volatiles
were removed in vacuo, the residue was dissolved in DMSO and
purified by reverse phase HPLC. The product fractions were
lyophilized directly to
N-(4-((1R,3R,4S,5S)-3-amino-4-(2-hydroxyethylsulfonyl)-5-methylcyclohexyl-
)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 44%
yield. LC/MS (m/z)=549.6 (MH.sup.+), R.sub.t=0.58 min. .sup.1H NMR
(400 MHz, <dmso>) .delta. ppm 10.51 (s, 1H), 8.53 (s, 1H),
8.51 (d, J=4.0, 1H), 8.33 (dd, J=8.0, 4.0, 1H), 8.20 (t, J=8.0,
1H), 8.09 (broad doublet, J=4.0, 2H), 7.65-7.75 (m, 1H), 7.36 (t,
J=8.0, 2H), 7.30 (d, J=8.0, 1H), 3.84-4.02 (m, 3H), 3.58-3.68 (m,
1H), 3.43-3.53 (m, 1H), 3.28-3.36 (m, 1H), 3.04-3.14 (m, 1H),
1.92-2.18 (m, 4H), 1.56-1.63 (m, 1H), 1.24 (d, J=8.0, 3H).
Synthesis of
S-(1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl)-
-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl
ethanethioate
##STR00135##
[0390] To a solution of
(1S,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl)-5-
-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl
methanesulfonate (1.0 equiv.) in DMF (0.25 M) was added KSAc (6.0
equiv.). The capped solution was left stirring at rt for 20 hrs.
The solution was partitioned between EtOAc and H.sub.2O. The
organic layer was washed with H.sub.2O, Na.sub.2CO.sub.3(sat.),
NaCl.sub.(sat.), dried over MgSO.sub.4, filtered, concentrated and
purified by ISCO SiO.sub.2 chromatography to yield
S-(1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl)-
-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl
ethanethioate in 50% yield. LC/MS (m/z)=615.2 (MH.sup.+),
R.sub.t=0.96 min.
Synthesis of tert-butyl
(1R,2R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylthio)cyclohexylcarbamate
##STR00136##
[0392] To a solution of
S-(1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl)-
-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl
ethanethioate (1.0 equiv.) in MeOH (0.07 M) was added
K.sub.2CO.sub.3 (3.0 equiv.). The heterogeneous solution was capped
and left stirring at rt for 1 hr. Methyl iodide (1.5 equiv.) was
added and stirred at rt for 10 min. The volatiles were removed in
vacuo and the residue was partitioned between EtOAc and H.sub.2O.
The organic layer was washed with NaCl.sub.(sat.), dried over
MgSO.sub.4, filtered, concentrated and purified by ISCO SiO.sub.2
chromatography to yield tert-butyl
(1R,2R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylthio)cyclohexylcarbamate in 72% yield. LC/MS
(m/z)=587.2 (MH.sup.+), R.sub.t=0.96 min.
Synthesis of
N-(4-((1R,3R,4R,5S)-3-amino-5-methyl-4-(methylthio)cyclohexyl)pyridin-3-y-
l)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00137##
[0394] To a solution of HCl (30.0 equiv.) in dioxane was added to
tert-butyl
(1R,2R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylthio)cyclohexylcarbamate (1.0 equiv.). The
solution was capped and left standing at rt for 1 hr. The volatiles
were removed in vacuo, dissolved in DMSO and purified by reverse
phase HPLC to yield
N-(4-((1R,3R,4R,5S)-3-amino-5-methyl-4-(methylthio)cyclohexyl)pyridin-3-y-
l)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 43% yield. LC/MS
(m/z)=487.2 (MH.sup.+), R.sub.t=0.65 min.
Synthesis of
N-(4-((1R,3R,4R,5S)-3-amino-5-methyl-4-((S)-methylsulfinyl)cyclohexyl)pyr-
idin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide and
N-(4-((1R,3R,4R,5S)-3-amino-5-methyl-4-((R)-methylsulfinyl)cyclohexyl)pyr-
idin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00138##
[0396] To a solution of tert-butyl
(1R,2R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylthio)cyclohexylcarbamate (1.0 equiv.) in THF
(0.05 M) in a 0.degree. C. bath was added the oxone (1.0 equiv.) as
a solution in H.sub.2O. The solution was left stirring at 0.degree.
C. for 5 mins. The solution was diluted with EtOAc, washed with
H.sub.2O, NaCl.sub.(sat.), dried over MgSO.sub.4, filtered,
concentrated to yield Boc protected product. The material was
treated with 25% TFA/CH.sub.2Cl.sub.2 for 30 minutes, at which time
the volatiles were removed in vacuo, the residue was dissolved in
DMSO and purified by reverse phase HPLC. The product fractions were
lyophilized directly to yield
N-(4-((1R,3R,4R,5S)-3-amino-5-methyl-4-((S)-methylsulfinyl)cyclohex-
yl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 33%
yield, LC/MS=503.2 (MH.sup.+), Rt=0.58 min; and
N-(4-((1R,3R,4R,5S)-3-amino-5-methyl-4-((R)-methylsulfinyl)cyclohexyl)pyr-
idin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 37%
yield. LC/MS (m/z)=503.2 (MH'), R.sub.t=0.60 min.
Synthesis of
N-(4-((1R,3R,4R,5S)-3-amino-5-methyl-4-(methylsulfonyl)cyclohexyl)pyridin-
-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00139##
[0398] To a solution of tert-butyl
(1R,2R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylthio)cyclohexylcarbamate (1.0 equiv.) in THF
(0.04 M) in a 0.degree. C. bath was added the oxone (2.0 equiv.) as
a solution in H.sub.2O. The solution was left stirring at rt for 5
hrs. The solution was diluted with EtOAc, washed with H.sub.2O,
NaCl.sub.(sat.), dried over MgSO.sub.4, filtered, concentrated to
yield Boc protected product. The material was treated with 25%
TFA/CH.sub.2Cl.sub.2 for 30 minutes, at which time the volatiles
were removed in vacuo, the residue was dissolved in DMSO and
purified by reverse phase HPLC. The product fractions were
lyophilized directly to yield
N-(4-((1R,3R,4R,5S)-3-amino-5-methyl-4-(methylsulfonyl)cyclohexyl)pyridin-
-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 68% yield.
LC/MS (m/z)=519.2 (MH.sup.+), R.sub.t=0.60 min. .sup.1H NMR (400
MHz, <dmso>) .delta. ppm 10.44 (s, 1H), 8.60 (d, J=8.0, 1H),
8.48 (d, J=4.0, 1H), 8.34 (dd, J=8.0, 4.0, 1H), 8.20 (dd, J=8.0,
8.0, 1H), 8.00 (dd, J=16.0, 4.0, 1H), 7.67-7.74 (m, 1H), 7.34-7.38
(m, 3H), 3.14 (s, 3H), 3.02-3.12 (m, 2H), 2.18-2.24 (m, 1H),
1.84-1.96 (m, 3H), 1.62-1.72 (m, 1H), 1.38-1.48 (m, 1H), 1.18 (d,
J=4.0, 3H).
Synthesis of (+/-)-tert-butyl
((1R,2R,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-hydroxy-3-
-methylcyclohexyl)carbamate
##STR00140##
[0400] A solution of
(+/-)-(1R,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-(tert-butoxycarbonylamino)--
6-methylcyclohexyl acetate (1.0 equiv.) and Boc.sub.2O (2.1 equiv.)
in dioxane (0.34 M) was submerged in an 120.degree. C. oil bath,
fitted with a condenser and left stirring under Ar for 6 hrs. The
reaction was cooled to rt and the volatiles were removed in vacuo.
The residue was dissolved in EtOH (0.34 M), K.sub.2CO.sub.3 (10.0
equiv.) was added, a refluxing head was attached and the
heterogeneous solution was submerged in an 50.degree. C. oil bath
and left stirring for 24 hrs. The reaction was cooled to rt. The
volatiles were removed in vacuo and the residue was partitioned
between EtOAc and H.sub.2O. The organic layer was washed with
Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.), dried over MgSO.sub.4,
filtered, and concentrated. The residue was dissolved in
CH.sub.2Cl.sub.2/Heptane and left standing. The solid that form was
sonciated, filtered, rinsed with CH.sub.2Cl.sub.2 and pumped on to
yield (+/-)-tert-butyl
((1R,2R,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-hydroxy-3-
-methylcyclohexyl)carbamate in 85% yield. LC/MS (m/z)=422.3
(MH.sup.+), R.sub.t=0.65 min.
Synthesis of
(+/-)-(1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(tert-butoxycarbony-
lamino)pyridin-4-yl)-6-methylcyclohexyl methanesulfonate
##STR00141##
[0402] To a solution of (+/-)-tert-butyl
((1R,2R,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-hydroxy-3-
-methylcyclohexyl)carbamate (1.0 equiv.) in pyridine (0.17 M) was
added MsCl (5.0 equiv.). The capped solution was stirred for 5
minutes and then the homogeneous solution was left standing at rt
for 16 hrs. The volatiles were removed in vacuo and the residue was
partitioned between EtOAc and H.sub.2O. The organic layer was
washed with H.sub.2O, 10% CuSO.sub.4, H.sub.2O,
Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.), dried over MgSO.sub.4,
filtered, concentrated and purified by ISCO SiO.sub.2
chromatography to yield
(+/-)-(1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(tert-butoxycarbony-
lamino)pyridin-4-yl)-6-methylcyclohexyl methanesulfonate in 59%
yield. LC/MS (m/z)=500.3 (MH.sup.+), R.sub.t=0.74 min.
Synthesis of
(+/-)S-((1S,2R,4R,6S)-2-((tert-butoxycarbonyl)amino)-4-(3-((tert-butoxyca-
rbonyl)amino)pyridin-4-yl)-6-methylcyclohexyl)ethanethioate
##STR00142##
[0404] To a solution of
(+/-)-(1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(tert-butoxycarbony-
lamino)pyridin-4-yl)-6-methylcyclohexyl methanesulfonate (1.0
equiv.) in DMF (0.25 M) was added potassium thioacetate (6.0
equiv.). The mixture was stirred for 6 hours in a 60.degree. C.
bath under Ar. Upon cooling and the residue was partitioned between
EtOAc and H.sub.2O. The organic layer was washed with H.sub.2O
(3.times.), Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.), dried over
MgSO.sub.4, filtered, concentrated and purified by ISCO SiO.sub.2
chromatography to yield (+/-)S-((1
S,2R,4R,6S)-2-((tert-butoxycarbonyl)amino)-4-(3-((tert-butoxycarbonyl)ami-
no)pyridin-4-yl)-6-methylcyclohexyl) ethanethioate in 87% yield.
LC/MS (m/z)=480.3 (MH.sup.+), R.sub.t=0.82 min.
Synthesis of (+/-)tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-3-methyl-2-(methylsulfonyl)cyclohe-
xyl)carbamate
##STR00143##
[0406] To a solution of
(+/-)S-((1S,2R,4R,6S)-2-((tert-butoxycarbonyl)amino)-4-(3-((tert-butoxyca-
rbonyl)amino)pyridin-4-yl)-6-methylcyclohexyl) ethanethioate (1.0
equiv.) in MeOH (0.09 M) was added potassium carbonate (3.0
equiv.). The mixture was stirred for 15 minutes at which time
methyl iodide (1.1 eq.) was added and the solution was stirred at
rt for 15 minutes. The volatiles were removed in vacuo and the
residue was partitioned between EtOAc and H.sub.2O. The organic
layer was washed with H.sub.2O, NaCl.sub.(sat.), dried over
MgSO.sub.4, filtered, concentrated and purified by ISCO SiO.sub.2
chromatography to yield the methyl sulfide product in 99% yield.
LC/MS (m/z)=452.3 (MH.sup.+), R.sub.t=0.87 min. To a solution of
methyl sulfide (1.0 eq.) in THF (0.05 M) at rt was added an aqueous
solution of oxone (2.2 eq.) dropwise over 10 minutes. After
stirring at rt for 1 hour the solution was partitioned between
EtOAc and H.sub.2O. The organic layer was washed with H.sub.2O,
NaCl.sub.(sat.), dried over MgSO.sub.4, filtered, concentrated to
yield the bis Boc protected methyl sulfone product in 95% yield.
LC/MS (m/z)=484.2 (MH.sup.+), R.sub.t=0.77 min. The bis boc
protected cyclohexyl sulfone (1.0 equiv) was treated with 4M HCl in
dioxane for 3 hours to removed both Boc groups. Upon removal of the
volatiles in vacuo, the residue was suspended in 1:1
dioxane/Na.sub.2CO.sub.3(sat.) and
N-(tert-Butoxycarbonyloxy)succinimide (1.2 eq.) was added. After
stirring for 1 hour, additional
N-(tert-Butoxycarbonyloxy)succinimide (1.2 eq.) was added. After
stirring for an additional 2 hours the solution was extracted with
CH.sub.2Cl.sub.2, the combined organic layers were washed with
H.sub.2O, NaCl.sub.(sat.), dried over MgSO.sub.4, filtered,
concentrated and purified by ISCO SiO.sub.2 chromatography to yield
the (+/-)tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-3-methyl-2-(methylsulfonyl)cyclohe-
xyl)carbamate in 56% yield. LC/MS (m/z)=384.3 (MH.sup.+),
R.sub.t=0.57 min. Chiral purification was completed via SFC (20%
EtOH/80% n-heptanes, 20 mL/min, OJ column) to isolate the pure
enantiomers. The second peak correlated with tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-3-methyl-2-(methylsulfonyl)cyclohe-
xyl)carbamate.
Synthesis of tert-butyl
(1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-3-methyl-2-(1H-1,2,4-triazol-1-yl)c-
yclohexylcarbamate and tert-butyl
(1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-3-methyl-2-(1H-1,2,4-triazol-1-yl)c-
yclohexylcarbamate
##STR00144##
[0408] A solution of
(+/-)-(1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(tert-butoxycarbony-
lamino)pyridin-4-yl)-6-methylcyclohexyl methanesulfonate (1.0
equiv.), 1H-1,2,4-triazole (3.0 equiv.) and Cs.sub.2CO.sub.3 (3.0
equiv.) in DMF (0.15 M) was stirred at 80.degree. C. for 5 hrs. The
volatiles were removed in vacuo and the residue was partitioned
between EtOAc and H.sub.2O. The organic layer was washed with
Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.), dried over MgSO.sub.4,
filtered, concentrated and purified by RP HPLC, followed by free
basing by partitioning between an equal volume of EtOAc and
Na.sub.2CO.sub.3, separating, washing with NaCl.sub.(sat.), drying
over MgSO.sub.4, filtering, concentrating. Purification was
completed via SFC (20% MeOH, 100 mL/min, AD column) to yield
tert-butyl
(1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-3-methyl-2-(1H-1,2,4-triazol-1-yl)c-
yclohexylcarbamate (19% yield, 99% ee) and tert-butyl
(1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-3-methyl-2-(1H-1,2,4-triazol-1-yl)c-
yclohexylcarbamate (19% yield, 99% ee). LC/MS (m/z)=373.3
(MH.sup.+), R.sub.t=0.54 min.
Synthesis of tert-butyl
tert-butoxycarbonyl(4-((3R,4R,5S)-3-((tert-butoxycarbonyl)amino)-4-((tert-
-butyldimethylsilyl)oxy)-5-methylpiperidin-1-yl)pyridin-3-yl)carbamate
##STR00145##
[0410] To a solution of tert-butyl
(3R,4R,5S)-1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)-5-methy-
lpiperidin-3-ylcarbamate (1.0 equiv.) in CH.sub.2Cl.sub.2 (0.50 M)
at RT was added Boc.sub.2O (6.0 equiv.), followed by DMAP (2.0
equiv.). The resulting mixture was stirred at RT for 16 hrs. The
reaction mixture was then diluted with EtOAc and water. The organic
layer was washed with Brine, dried over MgSO.sub.4, concentrated
and purified by flash column chromatography to yield tert-butyl
tert-butoxycarbonyl(4-((3R,4R,5S)-3-((tert-butoxycarbonyl)amino)-4-((tert-
-butyldimethylsilyl)oxy)-5-methylpiperidin-1-yl)pyridin-3-yl)carbamate
in 57% yield. LC/MS (m/z)=637.3 (MH.sup.+), R.sub.t=1.17 min.
Synthesis of tert-butyl
tert-butoxycarbonyl(4-((3R,4R,5S)-3-((tert-butoxycarbonyl)amino)-4-hydrox-
y-5-methylpiperidin-1-yl)pyridin-3-yl)carbamate
##STR00146##
[0412] To a solution of tert-butyl
tert-butoxycarbonyl(4-((3R,4R,5S)-3-((tert-butoxycarbonyl)amino)-4-((tert-
-butyldimethylsilyl)oxy)-5-methylpiperidin-1-yl)pyridin-3-yl)carbamate
(1.0 equiv.) in THF (0.20 M) at RT was added TBAF (1.0 equiv.). The
resulting mixture was stirred at rt for 4 hrs. The reaction mixture
was then diluted with EtOAc and water. The organic layer was washed
with Brine, dried over MgSO.sub.4, concentrated and purified by
flash column chromatography to yield tert-butyl
tert-butoxycarbonyl(4-((3R,4R,5S)-3-((tert-butoxycarbonyl)amino)-4-hydrox-
y-5-methylpiperidin-1-yl)pyridin-3-yl)carbamate in 87% yield. LC/MS
(m/z)=523.4 (MH.sup.+), R.sub.t=0.72 min. .sup.1H NMR (400 MHz,
<cdcl3>) .delta. ppm 1.03 (d, J=6.65 Hz, 3H), 1.34-1.51 (m,
54H), 1.72-1.87 (m, 1H), 2.05 (s, 1H), 2.46-2.57 (m, 1H), 2.69 (t,
J=11.35 Hz, 1H), 2.78-2.94 (m, 1H), 3.00-3.14 (m, 1H), 3.45 (d,
J=12.52 Hz, 1H), 3.53-3.76 (m, 1H), 4.63 (d, J=6.26 Hz, 1H), 6.88
(d, J=5.48 Hz, 3H), 8.13 (s, 3H), 8.26-8.36 (m, 3H).
Synthesis of
(3R,4R,5S)-1-(3-(bis(tert-butoxycarbonyl)amino)pyridin-4-yl)-3-((tert-but-
oxycarbonyl)amino)-5-methylpiperidin-4-yl methanesulfonate
##STR00147##
[0414] To a solution of tert-butyl
tert-butoxycarbonyl(4-((3R,4R,5S)-3-((tert-butoxycarbonyl)amino)-4-hydrox-
y-5-methylpiperidin-1-yl)pyridin-3-yl)carbamate (1.0 equiv.) in DCM
(0.20 M) was added TEA (1.7 equiv.), followed by MsCl (1.3 equiv.).
The capped solution was stirred at rt for 90 mins. The reaction
mixture was quenched with NaHCO.sub.3(sat.), and extracted with
EtOAc. The organic layer was washed with NaCl.sub.(sat.), dried
over MgSO.sub.4, filtered, concentrated to yield
(3R,4R,5S)-1-(3-(bis(tert-butoxycarbonyl)amino)pyridin-4-yl)-3-((tert-but-
oxycarbonyl)amino)-5-methylpiperidin-4-yl methanesulfonate in 99%
yield. LC/MS (m/z)=601.3 (MH.sup.+), R.sub.t=0.83 min.
Synthesis of tert-butyl
(3R,4S,5S)-1-(3-aminopyridin-4-yl)-5-methyl-4-(1H-1,2,4-triazol-1-yl)pipe-
ridin-3-ylcarbamate
##STR00148##
[0416] A solution of
(3R,4R,5S)-3-(tert-butoxycarbonylamino)-1-(3-(tert-butoxycarbonylamino)py-
ridin-4-yl)-5-methylpiperidin-4-yl methanesulfonate (1.0 equiv.),
1H-1,2,4-triazole (3.0 equiv.) and Cs.sub.2CO.sub.3 (3.0 equiv.) in
DMF (0.17 M) was stirred at 90.degree. C. for 3 hrs. The mixture
was diluted with DMF, filtered and purified by RP HPLC, followed by
free basing by partitioning between an equal volume of EtOAc and
Na.sub.2CO.sub.3, separating, washing with NaCl.sub.(sat.), drying
over MgSO.sub.4, filtering, concentrating to yield tert-butyl
(3R,4S,5S)-1-(3-aminopyridin-4-yl)-5-methyl-4-(1H-1,2,4-triazol-1-yl)pipe-
ridin-3-ylcarbamate in 8% yield. LC/MS (m/z)=374.3 (MH.sup.+),
R.sub.t=0.53 min.
Synthesis of tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-3-methyl-2-(2-oxopyridin-1(2H)-yl)-
cyclohexyl)carbamate, tert-butyl
((1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-3-methyl-2-(2-oxopyridin-1(2H)-yl)-
cyclohexyl)carbamate, tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-3-methyl-2-(pyridin-2-yloxy)cycloh-
exyl)carbamate and tert-butyl
(1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-3-methyl-2-(pyridin-2-yloxy)cyclohe-
xyl)carbamate
##STR00149##
[0418] To a solution of
(1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(tert-butoxycarbonylamino-
)pyridin-4-yl)-6-methylcyclohexyl methanesulfonate (1.0 equiv.)
Cs.sub.2CO.sub.3 (3.0 equiv.) in DMF (0.15 M) was added
pyridin-2-ol (1.0 equiv). The mixture was stirred at 70.degree. C.
for 5 hrs. The reaction was worked up with EtOAc and Brine. The
organic layer was concentrated and was treated with 4N HCl (30.0
equiv.) in Dioxane for 2 hrs at which time the volatiles were
removed in vacuo. The redisue was dissolved in THF (0.15 M) and
tert-butyl 2,5-dioxopyrrolidin-1-yl carbonate (1.5 equiv.) was
added, followed by DIEA (3.0 equiv.). After stirring at rt for 3
hrs, the reaction was quenched with sat. NaHCO.sub.3 and extracted
with EtOAc. The organic layer was washed with Brine, dried over
Na.sub.2SO.sub.4 and concentrated. The crude was purified by prep
HPLC to yield two major peaks. The fractions of the first product
peak was combined and neutralized with sat. NaHCO.sub.3 and
extracted with EtOAc. The organic layer was washed with Brine,
dried over Na.sub.2SO.sub.4 and concentrated. Purification was
completed via SFC (20% (MeOH with 10% DEA), 100 mL/min, AD column)
to yield tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-3-methyl-2-(2-oxopyridin-1(2H)-yl)-
cyclohexyl)carbamate (8% yield, 99% ee) and tert-butyl
((1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-3-methyl-2-(2-oxopyridin-1(2H)-yl)-
cyclohexyl)carbamate (9% yield, 99% ee). LC/MS (m/z)=399.2
(MH.sup.+), R.sub.t=0.60 min. .sup.1H NMR (400 MHz, <dmso>)
.delta. ppm 0.72 (s, 3H), 1.30 (s, 9H), 1.63-1.81 (m, 2H),
1.89-2.00 (m, 2H), 2.02-2.18 (m, 2H), 3.04-3.13 (m, 1H), 4.03-4.12
(m, 1H), 4.91-5.02 (m, 1H), 5.04-5.13 (m, 2H), 6.17-6.26 (m, 1H),
6.31-6.41 (m, 1H), 6.98-7.07 (m, 1H), 7.32-7.41 (m, 2H), 7.68-7.74
(m, 1H), 7.74-7.82 (m, 1H), 7.85-7.90 (m, 1H). The fractions of the
second product peak was combined and neutralized with sat.
NaHCO.sub.3 and extracted with EtOAc. The organic layer was washed
with Brine, dried over Na.sub.2SO.sub.4 and concentrated.
Purification was completed via SFC (20% (MeOH with 10% DEA), 100
mL/min, AD column) to yield tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-3-methyl-2-(pyridin-2-yloxy)cycloh-
exyl)carbamate (14% yield, 99% ee) and tert-butyl
((1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-3-methyl-2-(pyridin-2-yloxy)cycloh-
exyl)carbamate (15% yield, 99% ee). LC/MS (m/z)=399.2 (MH.sup.+),
R.sub.t=0.65 min. .sup.1H NMR (400 MHz, <cdcl3>) .delta. ppm
0.95 (d, J=6.65 Hz, 3H), 1.35-1.78 (m, 12H), 1.97-2.08 (m, 2H),
2.76 (t, J=11.93 Hz, 1H), 3.72 (br. s., 2H), 3.88 (d, J=4.70 Hz,
1H), 5.43 (d, J=7.43 Hz, 1H), 5.59 (br. s., 1H), 6.79-6.94 (m, 2H),
7.08 (d, J=5.09 Hz, 1H), 7.56-7.65 (m, 1H), 7.98-8.17 (m, 3H).
Synthesis of tert-butyl
((1R,2S,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-azido-3-m-
ethylcyclohexyl)carbamate and tert-butyl
((1S,2R,3R,5S)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-azido-3-m-
ethylcyclohexyl)carbamate
##STR00150##
[0420] To a solution of
(+/-)-(1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(tert-butoxycarbony-
lamino)pyridin-4-yl)-6-methylcyclohexyl methanesulfonate (1.0
equiv.) in DMF (0.13 M) was added NaN.sub.3 (1.0 equiv.). The
solution was submerged in an 80.degree. C. oil bath and left
stirring under Ar for 16 hrs. The solution was cooled to rt and
left stirring under Ar overnight. The solution was partitioned
between EtOAc and H.sub.2O. The organic layer was washed with
Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.), dried over MgSO.sub.4,
filtered, concentrated and purified by ISCO SiO.sub.2
chromatography. Purification was completed via SFC (15% IPA, 100
mL/min, IA column) to yield tert-butyl
((1R,2S,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-azido-3-m-
ethylcyclohexyl)carbamate (21% yield, 99% ee) and tert-butyl
((1S,2R,3R,5S)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-azido-3-m-
ethylcyclohexyl)carbamate (22% yield, 99% ee). LC/MS (m/z)=447.3
(MH.sup.+), R.sub.t=0.86 min.
Synthesis of tert-butyl
(1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-2-azido-3-methylcyclohexylcarbamate
##STR00151##
[0422] A solution of 4 M HCl in dioxane (30.0 equiv.) was added to
tert-butyl
((1R,2S,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-azido-3-m-
ethylcyclohexyl)carbamate (1.0 equiv.). The solution started to go
homogeneous for a few minutes, but then a ppt formed and the
solution went very thick. After sitting at rt for 1 hour, the
volatiles were removed in vacuo and the solid was pumped on for 5
minutes on the high vac. To the residue was added CH.sub.2Cl.sub.2
(0.15 M), TEA (5.0 equiv.) and Boc.sub.2O (1.0 equiv.). The
solution was left stirring at rt for 1 hr. The volatiles were
removed in vacuo and the residue was partitioned between EtOAc and
H.sub.2O. The organic layer was washed with Na.sub.2CO.sub.3(sat.),
NaCl.sub.(sat.), dried over MgSO.sub.4, filtered, concentrated and
purified by ISCO SiO.sub.2 chromatography to yield tert-butyl
(1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-2-azido-3-methylcyclohexylcarbamate
in 57% yield. LC/MS (m/z)=347.3 (MH.sup.+), R.sub.t=0.70 min.
.sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 1.13 (d, J=6.65 Hz,
3H), 1.43-1.58 (m, 11H), 1.79 (d, J=12.52 Hz, 1H), 1.95 (d, J=6.26
Hz, 1H), 2.60 (br. s., 1H), 3.61 (br. s., 2H), 3.77-3.91 (m, 2H),
4.78 (d, J=7.43 Hz, 1H), 6.96 (d, J=4.70 Hz, 1H), 7.97-8.07 (m,
2H).
Synthesis of tert-butyl
(1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-2-azido-3-methylcyclohexylcarbamate
##STR00152##
[0424] A solution of 4 M HCl in dioxane (30.0 equiv.) was added to
tert-butyl
((1S,2R,3R,5S)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-azido-3-m-
ethylcyclohexyl)carbamate (1.0 equiv.). The solution started to go
homogeneous for a few minutes, but then a ppt formed and the
solution went very thick. After sitting at rt for 1 hour, the
volatiles were removed in vacuo and the solid was pumped on for 5
minutes on the high vac. To the residue was added CH.sub.2Cl.sub.2
(0.15 M), TEA (5.0 equiv.) and Boc.sub.2O (1.0 equiv.). The
solution was left stirring at rt for 1 hr. The volatiles were
removed in vacuo and the residue was partitioned between EtOAc and
H.sub.2O. The organic layer was washed with Na.sub.2CO.sub.3(sat.),
NaCl.sub.(sat.), dried over MgSO.sub.4, filtered, concentrated to
yield tert-butyl
(1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-2-azido-3-methylcyclohexylcarbamate
in 98% yield. LC/MS (m/z)=347.3 (MH.sup.+), R.sub.t=0.71 min.
Synthesis of tert-butyl
(1S,2R,3R,5S)-2-azido-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate
##STR00153##
[0426] EDC (2.0 equiv.) and HOAt (2.0 equiv.) was added to a
solution of tert-butyl
(1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-2-azido-3-methylcyclohexyl
carbamate (1.0 equiv.) and 6-(2,6-difluorophenyl)-5-fluoropicolinic
acid (1.5 equiv.) in DMF (0.08 M). The mixture was stirred at
ambient temperature overnight. The reaction mixture was diluted
with water and extracted with ethyl acetate. The combined extracts
were washed sequentially with 1M aqueous sodium hydroxide and
brine, dried over sodium sulfate, filtered, concentrated and
purified by ISCO SiO.sub.2 chromatography to yield tert-butyl
(1S,2R,3R,5S)-2-azido-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexyl carbamate in 31% yield. LC/MS
(m/z)=582.3 (MH.sup.+), R.sub.t=1.00 min.
Synthesis of tert-butyl
(1R,2S,3S,5R)-2-azido-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate
##STR00154##
[0428] EDC (2.0 equiv.) and HOAt (2.0 equiv.) was added to a
solution of tert-butyl
(1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-2-azido-3-methylcyclohexylcarbamate
(1.0 equiv.) and 6-(2,6-difluorophenyl)-5-fluoropicolinic acid (1.5
equiv.) in DMF (0.08 M). The mixture was stirred at ambient
temperature overnight. The reaction mixture was diluted with water
and extracted with ethyl acetate. The combined extracts were washed
sequentially with 1M aqueous sodium hydroxide and brine, dried over
sodium sulfate, filtered, concentrated and purified by ISCO
SiO.sub.2 chromatography to yield tert-butyl
(1R,2S,3S,5R)-2-azido-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate 59% yield. LC/MS
(m/z)=582.3 (MH.sup.+), R.sub.t=0.97 min.
Synthesis of tert-butyl
(1R,2S,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate
##STR00155##
[0430] Degass a solution of tert-butyl
(1R,2S,3S,5R)-2-azido-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate (1.0 equiv.) in MeOH/EtOAc
(3/1, 0.04 M). To this solution was added Pd/C (0.2 equiv.) and
purge with Ar and H.sub.2. The mixture was stirred under H.sub.2
for 16 hrs. Filter the mixture over cetlite and wash the cake with
MeOH. Concentrate the filtrate to yield tert-butyl
(1R,2S,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate in 100% yield). LC/MS
(m/z)=556.3 (MH.sup.+), R.sub.t=0.73 min.
Method 4
Synthesis of ethyl
(1S,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexylcarbamate
##STR00156##
[0432] To a solution of tert-butyl
(1R,2S,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate (1.0 equiv.) in
CH.sub.2Cl.sub.2 (0.03 M) at 0.degree. C. was added DIEA (3.0
equiv.) and then ETHYL CHLOFORMATE (1.0 equiv.). The homogeneous
solution was left standing at 0.degree. C. at for 4 hrs. Neutralize
the reaction with sat. NaHCO.sub.3 solution. The solution was
partitioned between EtOAc and H.sub.2O. The organic layer was
washed with Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.), dried over
MgSO.sub.4, filtered, concentrated and purified by ISCO SiO.sub.2
chromatography. The Boc protected product was treated with 25%
TFA/CH.sub.2Cl.sub.2 for 20 minutes at which time the volatiles
were removed in vacuo and the residue was dissolved in DMSO and
purified by RP-HPLC. The product fractions were lyophilized
directly to yield ethyl
(1S,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexylcarbamate in 14% yield. LC/MS
(m/z)=528.2 (MH.sup.+), R.sub.t=0.65 min.
Synthesis of isopropyl
(1S,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexylcarbamate
##STR00157##
[0434] Method 4 was followed using tert-butyl
(1R,2S,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexyl carbamate and isopropyl
carbonochloridate to give isopropyl
(1S,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methyl cyclohexylcarbamate in 6% yield. LC/MS
(m/z)=542.3 (MH.sup.+), R.sub.t=0.68 min.
Synthesis of
N-(4-((1R,3R,4S,5S)-3-amino-5-methyl-4-propionamidocyclohexyl)pyridin-3-y-
l)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00158##
[0436] Method 4 was followed using tert-butyl
(1R,2S,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexyl carbamate, TEA and propionyl
chloride to give
N-(4-((1R,3R,4S,5S)-3-amino-5-methyl-4-propionamidocyclohexyl)pyridi-
n-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 8% yield.
LC/MS (m/z)=512.1 (MH.sup.+), R.sub.t=0.62 min.
Synthesis of
N-(4-((1R,3R,4S,5S)-3-amino-4-isobutyramido-5-methylcyclohexyl)pyridin-3--
yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00159##
[0438] Method 4 was followed using tert-butyl
(1R,2S,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexyl carbamate, TEA and isobutyryl
chloride to give
N-(4-((1R,3R,4S,5S)-3-amino-4-isobutyramido-5-methylcyclohexyl)pyrid-
in-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 16% yield.
LC/MS (m/z)=526.3 (MH.sup.+), R.sub.t=0.66 min.
Synthesis of
N-(4-((1R,3R,4S,5S)-3-amino-4-(2-methoxyacetamido)-5-methylcyclohexyl)pyr-
idin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00160##
[0440] Method 4 was followed using tert-butyl
(1R,2S,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexyl carbamate, TEA and 2-methoxyacetyl
chloride to give
N-(4-((1R,3R,4S,5S)-3-amino-4-(2-methoxyacetamido)-5-methylcyclohexyl)pyr-
idin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 24%
yield. LC/MS (m/z)=528.2 (MH.sup.+), R.sub.t=0.62 min.
Synthesis of
N-(4-((1R,3R,4R,5S)-4-acetamido-3-amino-5-methylcyclohexyl)pyridin-3-yl)--
6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00161##
[0442] Method 4 was followed using tert-butyl
((1R,2R,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)-
pyridin-4-yl)-3-methylcyclohexyl)carbamate and acetic anhydride to
give
N-(4-((1R,3R,4R,5S)-4-acetamido-3-amino-5-methylcyclohexyl)pyridin-3-yl)--
6-(2,6-difluorophenyl)-5-fluoropicolinamide in 13% yield. LC/MS
(m/z)=498.3 (MH.sup.+), R.sub.t=0.58 min.
Method 5
Synthesis of
N-(4-((1R,3R,4S,5S)-3-amino-5-methyl-4-(methylsulfonamido)cyclohexyl)pyri-
din-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide and
N-(4-((1S,3S,4R,5R)-3-amino-5-methyl-4-(methylsulfonamido)cyclohexyl)pyri-
din-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00162##
[0444] To a solution of (+/-)-tert-butyl
(1R,2S,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexyl carbamate (1.0 equiv.) in
CH.sub.2Cl.sub.2 (0.03 M) at 0.degree. C. was added DIEA (3.0
equiv.) and then METHANESULFONYL CHLORIDE (1.5 equiv.). The
homogeneous solution was left standing at 0.degree. C. at for 2
hrs. Neutralize the reaction with sat. NaHCO.sub.3 solution. The
solution was partitioned between EtOAc and H.sub.2O. The organic
layer was washed with Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.),
dried over MgSO.sub.4, filtered, concentrated and purified by ISCO
SiO.sub.2 chromatography. Purification was completed via SFC (MeOH,
100 mL/min, OD column) to yield tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylsulfonamido)cyclohexylcarbamate (14% yield,
99% ee) and tert-butyl
(1S,2R,3R,5S)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(methylsulfonamido)cyclohexylcarbamate (13% yield,
99% ee). LC/MS (m/z)=634.3 (MH.sup.+), R.sub.t=0.86 min. Each Boc
protected enantiomer was treated respectively with 25%
TFA/CH.sub.2Cl.sub.2 for 20 minutes at which time the volatiles
were removed in vacuo and the residue was dissolved in DMSO and
purified by RP-HPLC to yield
N-(4-((1R,3R,4S,5S)-3-amino-5-methyl-4-(methylsulfonamido)cyclohexyl)pyri-
din-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 94% yield;
N-(4-((1S,3S,4R,5R)-3-amino-5-methyl-4-(methylsulfonamido)cyclohexyl)pyri-
din-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 97% yield.
LC/MS (m/z)=534.2 (MH.sup.+), R.sub.t=0.59 min.
Synthesis of
N-(4-((1R,3R,4S,5S)-4-acetamido-3-amino-5-methylcyclohexyl)pyridin-3-yl)--
6-(2,6-difluorophenyl)-5-fluoropicolinamide and N-(4-((1
S,3S,4R,5R)-4-acetamido-3-amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-d-
ifluorophenyl)-5-fluoropicolinamide
##STR00163##
[0446] Method 5 was followed using (+/-)-tert-butyl
(1R,2S,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexyl carbamate and acetic anhydride to
give
N-(4-((1R,3R,4S,5S)-4-acetamido-3-amino-5-methylcyclohexyl)pyridin-3-yl)--
6-(2,6-difluorophenyl)-5-fluoropicolinamide in 20% yield;
N-(4-((1S,3S,4R,5R)-4-acetamido-3-amino-5-methylcyclohexyl)pyridin-3-yl)--
6-(2,6-difluorophenyl)-5-fluoropicolinamide in 25% yield. LC/MS
(m/z)=498.2 (MH.sup.+), R.sub.t=0.59 min.
Synthesis of methyl
(1R,2S,4S,6R)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexylcarbamate and methyl
(1S,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexylcarbamate
##STR00164##
[0448] Method 5 was followed using (+/-)-tert-butyl
(1R,2S,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexyl carbamate and methyl
carbonochloridate to give methyl
(1R,2S,4S,6R)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexylcarbamate in 9% yield; methyl
(1S,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-6-methylcyclohexylcarbamate in 10% yield. LC/MS
(m/z)=514.2 (MH.sup.+), R.sub.t=0.62 min.
Synthesis of
(+/-)-(1S,2R,6S)-2-((tert-butoxycarbonyl)amino)-6-methyl-4-(3-nitropyridi-
n-4-yl)cyclohex-3-en-1-yl methanesulfonate
##STR00165##
[0450] To a solution of (+/-)-tert-butyl
((1R,5S,6S)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-en-1-yl)-
carbamate (1.0 equiv.) in pyridine (0.20 M) was added MsCl (5.0
equiv.). The capped solution was stirred for 5 minutes and then the
homogeneous solution was left standing at rt for 16 hrs. The
volatiles were removed in vacuo and the residue was partitioned
between EtOAc and H.sub.2O. The organic layer was washed with 10%
CuSO.sub.4, H.sub.2O, Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.),
dried over MgSO.sub.4, filtered, concentrated and purified by ISCO
SiO.sub.2 chromatography to yield
(+/-)-(1S,2R,6S)-2-((tert-butoxycarbonyl)amino)-6-methyl-4-(3-nitropyridi-
n-4-yl)cyclohex-3-en-1-ylmethanesulfonate in 46% yield. LC/MS
(m/z)=428.2 (MH.sup.+), R.sub.t=0.89 min.
Synthesis of
(+/-)-(1S,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-((tert-butoxycarbonyl)amino-
)-6-methylcyclohexyl methanesulfonate
##STR00166##
[0452] Degass a solution of
(+/-)-(1S,2R,6S)-2-((tert-butoxycarbonyl)amino)-6-methyl-4-(3-nitropyridi-
n-4-yl)cyclohex-3-en-1-ylmethanesulfonate (1.0 equiv.) in Ethanol
(0.20 M). To this solution was added Pd/C (0.2 equiv.) and purge
with Ar and H.sub.2. The mixture was stirred under H.sub.2 for 16
hrs. Filter the mixture over cetlite and wash the cake with MeOH.
Concentrate the filtrate to yield
(+/-)-(1S,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-((tert-butoxycarbonyl)amino-
)-6-methylcyclohexyl methanesulfonate in 49% yield). LC/MS
(m/z)=400.3 (MH.sup.+), R.sub.t=0.62 min.
Synthesis of (+/-)-tert-butyl
((1R,2R,3S,5R)-5-(3-aminopyridin-4-yl)-2-azido-3-methylcyclohexyl)carbama-
te
##STR00167##
[0454] To a solution of
(+/-)-(1S,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-((tert-butoxycarbonyl)amino-
)-6-methylcyclohexyl methanesulfonate (1.0 equiv.) in DMF (0.20 M)
was added NaN.sub.3 (7.0 equiv.). The solution was submerged in a
70.degree. C. oil bath and left stirring under Ar for 4 hrs. The
solution was cooled to rt and left stirring under Ar overnight. The
solution was partitioned between EtOAc and H.sub.2O. The organic
layer was washed with Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.),
dried over MgSO.sub.4, filtered, concentrated to yield
(+/-)-tert-butyl
((1R,2R,3S,5R)-5-(3-aminopyridin-4-yl)-2-azido-3-methyl
cyclohexyl)carbamate in 87% yield. LC/MS (m/z)=347.3 (MH.sup.+),
R.sub.t=0.68 min.
Synthesis of (+/-)-tert-butyl
((1R,2R,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)-
pyridin-4-yl)-3-methylcyclohexyl)carbamate
##STR00168##
[0456] EDC (2.0 equiv.) and HOAt (2.0 equiv.) was added to a
solution of (+/-)-tert-butyl
((1R,2R,3S,5R)-5-(3-aminopyridin-4-yl)-2-azido-3-methyl
cyclohexyl)carbamate (1.0 equiv.) and
6-(2,6-difluorophenyl)-5-fluoropicolinic acid (1.5 equiv.) in DMF
(0.20 M). The mixture was stirred at ambient temperature overnight.
The reaction mixture was diluted with water and extracted with
ethyl acetate. The combined extracts were washed sequentially with
1M aqueous sodium hydroxide and brine, dried over sodium sulfate,
filtered, concentrated and purified by ISCO SiO.sub.2
chromatography. To a degassed a solution of the azide (1.0 equiv.)
in 2-propanol (0.10 M) was added Pd/C (0.2 equiv.). The mixture was
stirred under H.sub.2 for 48 hrs. Filter the mixture over cetlite
and wash the cake with MeOH. Concentrate the filtrate to yield
(+/-)-tert-butyl
((1R,2R,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)-
pyridin-4-yl)-3-methylcyclohexyl)carbamate in 35% yield). LC/MS
(m/z)=556.3 (MH.sup.+), R.sub.t=0.64 min.
Synthesis of
N-(4-((1R,3R,4R,5S)-3-amino-5-methyl-4-(methylsulfonamido)cyclohexyl)pyri-
din-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide and
N-(4-((1S,3S,4S,5R)-3-amino-5-methyl-4-(methylsulfonamido)cyclohexyl)pyri-
din-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00169##
[0458] Method 5 was followed using (+/-)-tert-butyl
(1R,2R,3S,5R)-2-amino-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate and methanesulfonyl
chloride to give
N-(4-((1R,3R,4R,5S)-3-amino-5-methyl-4-(methylsulfonamido)cyclohexyl-
)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 13%
yield;
N-(4-((1S,3S,4S,5R)-3-amino-5-methyl-4-(methylsulfonamido)cyclohexyl)pyri-
din-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 14% yield.
LC/MS (m/z)=534.2 (MH.sup.+), R.sub.t=0.58 min.
Synthesis of tert-butyl
(1R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-3-methyl-2-oxocyclohexylcarbamate
##STR00170##
[0460] To a solution of tert-butyl
(1R,2R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-2-hydroxy-3-methylcyclohexylcarbamate (1.0 equiv.) in DCM
(0.10 M) was added Dess-MartinPeriodinane (1.2 equiv.). The flask
was capped and the homogeneous solution was left stirring at rt for
3 hrs. The solution was partitioned between EtOAc and 1:1 10%
Na.sub.2S.sub.2O.sub.3/NaHCO.sub.3 (sat.). The organic layer was
washed with NaCl(sat.), dried over MgSO.sub.4, filtered and
concentrated, and purified by ISCO SiO.sub.2 chromatography to
yield tert-butyl
(1R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-3-methyl-2-oxocyclohexylcarbamate in 83% yield. LC/MS (m/z)=555.4
(MH.sup.+), R.sub.t=0.87 min.
Synthesis of N-(4-((1R,3R,5
S,E)-3-amino-4-(methoxyimino)-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-dif-
luorophenyl)-5-fluoropicolinamide and
N-(4-((1R,3R,5S,Z)-3-amino-4-(methoxyimino)-5-methylcyclohexyl)pyridin-3--
yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00171##
[0462] A solution of methoxylamine-HCl (1.0 equiv.) and tert-butyl
(1R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-3-methyl-2-oxocyclohexylcarbamate (1.0 equiv.) in EtOH/pyridine
(1/1, 0.01 M) was capped and left standing at rt for 16 hrs. The
volatiles were removed in vacuo and the residue was partitioned
between EtOAc and Na.sub.2CO.sub.3(sat.). The organic layer was
washed with NaCl.sub.(sat.), dried over MgSO.sub.4, filtered,
concentrated. The Boc group was removed with 25%
TFA/CH.sub.2Cl.sub.2. After 45 minutes, the volatiles were removed
in vacuo and the residue was pumped on for 5 minutes, dissolved in
CH.sub.2Cl.sub.2 and neutralized with TEA. The volatiles were
removed in vacuo and after pumping the residue was dissolved in
DMSO and purified by RP-HPLC to yield
N-(4-((1R,3R,5S,E)-3-amino-4-(methoxyimino)-5-methylcyclohexyl)pyridin-3--
yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 16% yield. LC/MS
(m/z)=484.2 (MH), R.sub.t=0.64 min. .sup.1H NMR (400 MHz, DMSO-d6)
.delta. ppm 10.49 (s, 1H), 8.57 (s, 1H), 8.47 (d, J=4.0, 1H), 8.35
(dd, J=8.0, 4.0, 1H), 8.25 (broad doublet, J=4.0, 2H), 8.20 (t,
J=8.0, 1H), 7.67-7.74 (m, 1H), 7.42 (d, J=8.0, 1H), 7.36 (t, J=8.0,
2H), 4.04-4.08 (m, 1H), 3.79 (s, 3H), 3.23-3.29 (m, 1H), 2.39-2.45
(m, 1H), 2.11 (d, J=8.0, 1H), 2.10 (d, J=8.0, 1H), 1.90 (q, J=12,
1H), 1.40 (q, J=12, 1H), 1.01 (d, J=4.0, 3H); and
N-(4-((1R,3R,5S,Z)-3-amino-4-(methoxyimino)-5-methylcyclohexyl)pyridin-3--
yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 27% yield. LC/MS
(m/z)=484.2 (MH.sup.+), R.sub.t=0.66 min. .sup.1H NMR (400 MHz,
DMSO-d6) .delta. ppm 10.48 (s, 1H), 8.57 (s, 1H), 8.47 (d, J=4.0,
1H), 8.35 (dd, J=8.0, 4.0, 1H), 8.20 (t, J=8.0, 1H), 8.08 (broad
singlet, 2H), 7.67-7.74 (m, 1H), 7.42 (d, J=8.0, 1H), 7.36 (t,
J=8.0, 2H), 3.88-3.92 (m, 1H), 3.80 (s, 3H), 3.22-3.28 (m, 1H),
2.51-2.58 (m, 1H), 2.25 (d, J=12.0, 1H), 1.86 (d, J=12.0, 1H), 1.70
(q, J=12, 1H), 1.62 (q, J=12, 1H), 1.34 (d, J=4.0, 3H).
Synthesis of
N-(4-((1R,3R,5S,Z)-3-amino-4-(hydroxyimino)-5-methylcyclohexyl)pyridin-3--
yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00172##
[0464] A solution of hydroxylamine-HCl (4.0 equiv.) and tert-butyl
(1R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-3-methyl-2-oxocyclohexylcarbamate (1.0 equiv.) in EtOH/pyridine
(1/1, 0.01 M) was capped and left standing at rt for 16 hrs. The
volatiles were removed in vacuo and the residue was partitioned
between EtOAc and Na.sub.2CO.sub.3(sat.). The organic layer was
washed with NaCl.sub.(sat.), dried over MgSO.sub.4, filtered,
concentrated. The Boc group was removed with 25%
TFA/CH.sub.2Cl.sub.2. After 45 minutes, the volatiles were removed
in vacuo and after pumping the residue was dissolved in DMSO and
purified by RP-HPLC to yield
N-(4-((1R,3R,5S,Z)-3-amino-4-(hydroxyimino)-5-methylcyclohexyl)pyridin-3--
yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in 14% yield. LC/MS
(m/z)=470.3 (MH.sup.+), R.sub.t=0.60 min. .sup.1H NMR (300 MHz,
DMSO-d6) .delta. ppm 10.94 (s, 1H), 10.49 (s, 1H), 8.60 (s, 1H),
8.48 (d, J=4.0, 1H), 8.35 (dd, J=8.0, 4.0, 1H), 8.20 (t, J=8.0, 1
H), 8.02 (broad doublet, J=4.0, 2H), 7.67-7.74 (m, 1H), 7.42 (d,
J=4.0, 1H), 7.36 (t, J=8.0, 2H), 4.24 (m, 1H), 3.82-3.86 (m, 1H),
3.21-3.27 (m, 1H), 2.50-2.55 (m, 1H), 2.24 (d, J=12.0, 1H), 1.86
(d, J=16.0, 1H), 1.68 (q, J=12.0, 1H), 1.59 (q, J=12.0, 1H), 1.40
(d, J=8.0, 3H).
Synthesis of
N-(4-((1R,3R,5S)-3-amino-5-methyl-4-oxocyclohexyl)pyridin-3-yl)-6-(2,6-di-
fluorophenyl)-5-fluoropicolinamide
##STR00173##
[0466] Tert-butyl
(1R,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-3-methyl-2-oxocyclohexylcarbamate was treated with 25%
TFA/CH.sub.2Cl.sub.2 for 30 minutes. The volatiles were removed in
vacuo, the residue was dissolved in DMSO and purified by reverse
phase HPLC to yield
N-(4-((1R,3R,5S)-3-amino-5-methyl-4-oxocyclohexyl)pyridin-3-yl)-6-(-
2,6-difluorophenyl)-5-fluoropicolinamide in 98% yield. LC/MS
(m/z)=455.1 (MH.sup.+), R.sub.t=0.57 min. .sup.1H NMR (300 MHz,
DMSO-d6) .delta. ppm 10.55 (s, 1H), 8.55 (s, 1H), 8.47 (d, J=4.0,
1H), 8.37 (dd, J=8.0, 4.0, 1H), 8.21 (t, J=8.0, 1H), 8.16 (broad
doublet, J=4.0, 2H), 7.67-7.74 (m, 1H), 7.40 (d, J=8.0, 1H), 7.36
(t, J=8.0, 2H), 4.20-4.26 (m, 1H), 3.50-3.70 (m, 2H), 2.76-2.82 (m,
1H), 2.49-2.54 (m, 1H), 2.32-2.36 (m, 1H), 2.16-2.18 (m, 1H), 1.91
(q, J=12, 1H), 1.65 (q, J=12, 1H), 0.97 (d, J=8.0, 3H).
Synthesis of (+/-)-tert-butyl
((1R,5S,6R)-6-methoxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-en-1-yl)-
carbamate
##STR00174##
[0468] (+/-)-Tert-butyl
(1R,5S,6R)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enylcarba-
mate (1.0 equiv.) was suspended in iodomethane (100.0 equiv.).
Silver oxide (6.0 equiv.) was added to the mixture and the reaction
vessel was wrapped in foil (kept dark) and allowed to stir
45.degree. C. for 10 hrs. The reaction was diluted with THF and
filtered through a pad of celite. The celite cake was further
washed with MeOH. The organics were concentrated and the crude was
taken up in DCM, washed with NaHCO.sub.3(aq.), dried over
Na.sub.2SO.sub.4, filtered and concentrated. The crude was loaded
onto silica gel and purified via ISCO to yield (+/-)-tert-butyl
((1R,5S,6R)-6-methoxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-en-1-yl)-
carbamate in 35% yield. LC/MS (m/z)=364.1 (MH.sup.+), R.sub.t=0.89
min.
Method 6
Synthesis of tert-butyl
((1S,2S,3R,5S)-5-(3-aminopyridin-4-yl)-2-methoxy-3-methylcyclohexyl)carba-
mate and tert-butyl
((1R,2R,3S,5R)-5-(3-aminopyridin-4-yl)-2-methoxy-3-methylcyclohexyl)carba-
mate
##STR00175##
[0470] To a solution of (+/-)-tert-butyl
((1R,5S,6R)-6-methoxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-en-1-yl)-
carbamate (1.0 equiv.) in degassed EtOH (0.10 M) was added Pd/C
(0.1 equiv.). The mixture was purged with H.sub.2, and allowed to
to stir under an atm of H.sub.2 overnight at RT. The reaction was
filtered through a pad of celite and the cake was washed with MeOH.
The organics were concentrated and purified by ISCO SiO.sub.2
chromatography. Purification was completed via SFC (30% MeOH, 100
mL/min, AD column) to yield tert-butyl
((1S,2S,3R,5S)-5-(3-aminopyridin-4-yl)-2-methoxy-3-methylcyclohexyl)carba-
mate (15% yield, 99% ee) and tert-butyl
((1R,2R,3S,5R)-5-(3-aminopyridin-4-yl)-2-methoxy-3-methylcyclohexyl)carba-
mate (12% yield, 99% ee). LC/MS (m/z)=336.3 (MH.sup.+),
R.sub.t=0.58 min.
Synthesis of (+/-)-tert-butyl
((1R,5S,6S)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-en-1-yl)-
carbamate
##STR00176##
[0472] To a solution of (+/-)-(3aR,7S,7aS)-tert-butyl
7-methyl-5-(3-nitropyridin-4-yl)-2-oxo-3a,6,7,7a-tetrahydrobenzo[d]oxazol-
e-3(2H)-carboxylate (1.0 equiv.) in THF (0.20 M) was added 2M LiOH
(3.0 equiv.) was added. The mixture was stirred overnight 20 hrs at
22.degree. C. The mixture was diluted with EtOAc and
NaHCO.sub.3(aq.). The layers were separated and the aqueous was
extracted with EtOAc. The combined organics were washed with brine,
dried over Na.sub.2SO.sub.4, filtered, and concentrated. The golden
foam was purified by ISCO chromatography to afford (+/-)-tert-butyl
((1R,5S,6S)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-en-1-yl)-
carbamate in 83% yield. LC/MS (m/z)=350.2 (MH.sup.+), R.sub.t=0.82
min.
Synthesis of (+/-)-tert-butyl
(1R,5S,6S)-6-(2-cyanoethoxy)-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-e-
nylcarbamate
##STR00177##
[0474] A mixture of (+/-)-Tert-butyl
(1R,5S,6S)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enylcarba-
mate (1.0 equiv.), acrylonitrile (30.0 equiv.) and CESIUM CARBONATE
(1.2 equiv.) in t-BuOH (0.57 M) was stirred at 35.degree. C. for 3
hrs. The reaction was cooled to room temperature, followed by the
addition of NaHCO.sub.3 (aq.) and water. The mixture was extracted
with EtOAc and the combined organics were dried over MgSO.sub.4,
filtered, and concentrated. The sample was purified by ISCO
SiO.sub.2 chromatography to yield (+/-)-tert-butyl
(1R,5S,6S)-6-(2-cyanoethoxy)-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-e-
nylcarbamate in 94% yield. LC/MS (m/z)=403.3 (MH.sup.+),
R.sub.t=0.92 min. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm
1.13 (d, J=6.46 Hz, 3H), 1.46 (s, 9H), 1.99-2.18 (m, 2H), 2.20-2.36
(m, 1H), 2.65 (t, J=6.06 Hz, 2H), 3.68 (br. s., 1H), 3.88 (t,
J=5.99 Hz, 2H), 4.51 (br. s., 1H), 4.99 (d, J=9.15 Hz, 1H), 5.39
(br. s., 1H), 7.25 (d, J=4.99 Hz, 1H), 8.73 (d, J=4.94 Hz, 1H),
9.10 (s, 1H).
Synthesis of tert-butyl
((1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-2-(2-cyanoethoxy)-3-methylcyclohex-
yl)carbamate and tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-2-(2-cyanoethoxy)-3-methylcyclohex-
yl)carbamate
##STR00178##
[0476] Method 6 was followed using (+/-)-tert-butyl
(1R,5S,6S)-6-(2-cyanoethoxy)-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-e-
nylcarbamate with SFC (15% EtOH, 100 mL/min, OJ column) to yield
tert-butyl
((1S,2R,3R,55)-5-(3-aminopyridin-4-yl)-2-(2-cyanoethoxy)-3-methylcyclohex-
yl)carbamate (31% yield, 99% ee) and tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-2-(2-cyanoethoxy)-3-methylcyclohex-
yl) carbamate (26% yield, 99% ee). LC/MS (m/z)=375.3 (MH.sup.+),
R.sub.t=0.65 min.
Synthesis of (+/-)-tert-butyl
((1R,5S,6S)-6-methoxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-en-1-yl)-
carbamate
##STR00179##
[0478] To a solution of (+/-)-tert-butyl
((1R,5S,6S)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-en-1-yl)-
carbamate(1.0 equiv.) in MeI (100.0 equiv.) was added Ag.sub.2O
(5.5 equiv.). A reflux condenser was attached and the heterogeneous
solution under Ar was submerged in a 50.degree. C. bath and the
reaction was gently refluxing for 6 hrs. The solids were filtered,
rinsed with CH.sub.2Cl.sub.2. The volatiles were removed in vacuo,
the residue was partitioned between CH.sub.2Cl.sub.2 and
NaHCO.sub.3(sat.). The organic layer was dried over MgSO.sub.4,
filtered, concentrated and purified by ISCO SiO.sub.2
chromatography to yield (+/-)-tert-butyl
((1R,5S,6S)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-en-1-yl)-
carbamate in 59% yield. LC/MS (m/z)=364.5 (MH.sup.+), Rt=1.02 min.
.sup.1H NMR (400 MHz, <cdcl3>) .delta. ppm 1.11 (d, J=6.65
Hz, 3H), 1.46 (s, 9H), 1.95-2.13 (m, 2H), 2.18-2.28 (m, 1H), 3.47
(d, J=3.52 Hz, 1H), 3.57 (s, 3H), 4.45 (d, J=7.83 Hz, 1H), 5.01 (d,
J=9.39 Hz, 1H), 5.44 (br. s., 1H), 7.24 (s, 1H), 8.71 (d, J=5.09
Hz, 1H), 9.08 (s, 1H).
Synthesis of tert-butyl
((1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-2-methoxy-3-methylcyclohexyl)carba-
mate and tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-2-methoxy-3-methylcyclohexyl)carba-
mate
##STR00180##
[0480] To a degassed solution of (+/-)-tert-butyl
((1R,5S,6S)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-en-1-yl)-
carbamate (1.0 equiv.) in i-PrOH (0.07 M) was added Pd/C (0.1
equiv.). The solution was degassed and purged to H.sub.2 and left
stirring under a balloon of H.sub.2 at rt for 16 hrs. The solution
was degassed and purged to Ar, diluted with CH.sub.2Cl.sub.2,
filtered through a pad of celite, concentrated and purified by ISCO
SiO.sub.2 chromatography. Purification was completed via SFC (20%
MeOH, 100 mL/min, AD column) to yield tert-butyl
((1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-2-methoxy-3-methylcyclohexyl)carba-
mate (42% yield, 99% ee) and tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-2-methoxy-3-methylcyclohexyl)carba-
mate (39% yield, 99% ee). LC/MS (m/z)=336.2 (MH.sup.+),
R.sub.t=0.67 min. .sup.1H NMR (400 MHz, <cdcl3>) .delta. ppm
1.08 (d, J=7.04 Hz, 3H), 1.43-1.49 (m, 10H), 1.52-1.64 (m, 2H),
1.70-1.81 (m, 2H), 2.52-2.64 (m, 1H), 3.39 (br. s., 1H), 3.52-3.57
(m, 3H), 3.62 (br. s., 2H), 3.66-3.75 (m, 1H), 4.75-4.87 (m, 1H),
6.98 (d, J=5.09 Hz, 1H), 7.95-8.05 (m, 2H).
Synthesis of (+/-)-tert-butyl
(1R,5S,6S)-6-ethoxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enylcarbam-
ate
##STR00181##
[0482] (+/-)-Tert-butyl
(1R,5S,6S)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enylcarba-
mate (1.0 equiv.) was suspended in iodoethane (100.0 equiv.).
Silver oxide (6.0 equiv.) was added to the mixture and the reaction
vessel was wrapped in foil (kept dark) and allowed to stir
55.degree. C. for 10 hrs. The reaction was diluted with THF and
filtered through a pad of celite. The celite cake was further
washed with MeOH. The organics were concentrated and the crude was
taken up in DCM, washed with NaHCO.sub.3 (aq), dried over
Na.sub.2SO.sub.4, filtered and concentrated. The crude was loaded
onto silica gel and purified via ISCO to yield (+/-)-tert-butyl
(1R,5S,6S)-6-ethoxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enylcarbam-
ate in 31% yield. LC/MS (m/z)=378.1 (MH.sup.+), R.sub.t=0.99
min.
Synthesis of tert-butyl
((1S,2R,3R,5S)-5-(3-aminopyridin-4-yl)-2-ethoxy-3-methylcyclohexyl)carbam-
ate and tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-2-ethoxy-3-methylcyclohexyl)carbam-
ate
##STR00182##
[0484] Method 6 was followed using (+/-)-tert-butyl
(1R,5S,6S)-6-ethoxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enylcarbam-
ate with Chiral HPLC (Heptane/EtOH=90/10, 20 mL/min, IC column) to
yield tert-butyl
((1S,2R,3R,55)-5-(3-aminopyridin-4-yl)-2-ethoxy-3-methylcyclohexyl)carbam-
ate (33% yield, 99% ee) and tert-butyl
((1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-2-ethoxy-3-methylcyclohexyl)carbam-
ate (28% yield, 99% ee). LC/MS (m/z)=350.2 (MH.sup.+), R.sub.t=0.72
min.
Synthesis of (+/-)-methyl
3-((1R,2R,6S)-2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-y-
l)cyclohex-3-enyloxy)propanoate
##STR00183##
[0486] A mixture of (+/-)-tert-butyl
(1R,5S,6R)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enylcarba-
mate (1.0 equiv.), methyl acrylate (30.0 equiv.) and CESIUM
CARBONATE (1.2 equiv.) in t-BuOH (0.38 M) was stirred at 35.degree.
C. for 16 hrs. The reaction was cooled to room temperature,
followed by the addition of NaHCO.sub.3(aq.) and water. The mixture
was extracted with EtOAc and the combined organics were dried over
MgSO.sub.4, filtered, and concentrated. The sample was purified by
ISCO chromatography to yield (+/-)-methyl
3-((1R,2R,6S)-2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-y-
l)cyclohex-3-enyloxy)propanoate in 48% yield. LC/MS (m/z)=436.1
(MH.sup.+), R.sub.t=0.91 min.
Synthesis of (+/-)-methyl
3-((1R,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-(tert-butoxycarbonylamino)-6-m-
ethylcyclohexyloxy)propanoate
##STR00184##
[0488] To a solution of (+/-)-methyl
3-((1R,2R,6S)-2-(tert-butoxycarbonylamino)-6-methyl-4-(3-nitropyridin-4-y-
l)cyclohex-3-enyloxy)propanoate (1.0 equiv.) in degassed EtOH (0.07
M) was added Pd/C (0.3 equiv.). The mixture was purged with
H.sub.2, and allowed to stir under H.sub.2 overnight at RT. The
reaction was filtered through a pad of celite and the cake was
washed with MeOH. The organics were concentrated and purified by
ISCO SiO.sub.2 chromatography to yield (+/-)-methyl
3-((1R,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-(tert-butoxycarbonylamino)-6-m-
ethylcyclohexyloxy)propanoate in 100% yield. LC/MS (m/z)=408.2
(MH.sup.+), R.sub.t=0.62 min.
Synthesis of methyl
3-((1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl-
)-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyloxy)propanoate
and methyl
3-(((1S,2S,4S,6R)-2-((tert-butoxycarbonyl)amino)-4-(3-(6-(2,6-difl-
uorophenyl)-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl)oxy)
propanoate
##STR00185##
[0490] EDC (2.0 equiv.) and HOAt (2.0 equiv.) was added to a
solution of (+/-)-methyl
3-((1R,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-(tert-butoxycarbonylamino)-6-m-
ethylcyclohexyloxy) propanoate (1.0 equiv.) and
6-(2,6-difluorophenyl)-5-fluoropicolinic acid (1.5 equiv.) in DMF
(0.08 M). The mixture was stirred at ambient temperature overnight.
The reaction mixture was diluted with water and extracted with
ethyl acetate. The combined extracts were washed sequentially with
1M aqueous sodium hydroxide and brine, dried over sodium sulfate,
filtered, concentrated and purified by ISCO SiO.sub.2
chromatography. Purification was completed via SFC (20% IPA, 20
mL/min, AD column) to yield methyl
3-((1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl-
)-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyloxy)propanoate
(22% yield, 99% ee) and methyl
3-(((1S,2S,4S,6R)-2-((tert-butoxycarbonyl)amino)-4-(3-(6-(2,6-difluorophe-
nyl)-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl)oxy)
propanoate (21% yield, 99% ee). LC/MS (m/z)=643.4 (MH.sup.+),
R.sub.t=0.92 min.
Synthesis of
3-((1R,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamid-
o)pyridin-4-yl)-6-methylcyclohexyloxy)propanoic acid
##STR00186##
[0492] To a solution of methyl
3-((1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(6-(2,6-difluorophenyl-
)-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyloxy)
propanoate (1.0 equiv.) was added 4 M HCl (40.0 equiv.) in dioxane.
After stirring at rt overnight, the mixture was concentrated and
dissolved in MeOH (0.05 M). LiOH (20.0 equiv.) was added. After
stirred 10 min at rt, the mixture was concentrated, neutralized
with HCl to PH 7 and extracted with EtOAc/t-Butanol (1/1). The
organic layer was wash with brine, dried over MgSO.sub.4, filtered,
and concentrated. The sample was purified by RP HPLC to yield
3-((1R,2R,4R,6S)-2-amino-4-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamid-
o)pyridin-4-yl)-6-methylcyclohexyloxy) propanoic acid in 34% yield.
LC/MS (m/z)=529.3 (MH.sup.+), R.sub.t=0.63 min.
Synthesis of (+/-)-tert-butyl
(1R,5S,6S)-5-methyl-6-(2-(methylsulfonyl)ethoxy)-3-(3-nitropyridin-4-yl)c-
yclohex-2-enylcarbamate
##STR00187##
[0494] A mixture of (+/-)-Tert-butyl
(1R,5S,6S)-6-hydroxy-5-methyl-3-(3-nitropyridin-4-yl)cyclohex-2-enylcarba-
mate (1.0 equiv.), methylsulfonylethene (30.0 equiv.) and CESIUM
CARBONATE (1.2 equiv.) in t-BuOH (0.22 M) was stirred at 22.degree.
C. for 5 hrs. The reaction was cooled to room temperature, followed
by the addition of NaHCO.sub.3(aq.) and water. The mixture was
extracted with EtOAc and the combined organics were dried over
MgSO.sub.4, filtered, and concentrated. The sample was purified by
ISCO chromatography to yield (+/-)-tert-butyl
(1R,5S,6S)-5-methyl-6-(2-(methylsulfonyl)ethoxy)-3-(3-nitropyridin-4-yl)c-
yclohex-2-enylcarbamate in 84% yield. LC/MS (m/z)=456.2 (MH.sup.+),
R.sub.t=0.87 min. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm
1.10 (d, J=6.60 Hz, 3H), 1.47 (s, 9H), 1.92-2.16 (m, 2H), 2.17-2.32
(m, 1H), 2.93-3.00 (m, 1H), 3.09 (s, 3H), 3.18 (d, J=14.87 Hz, 1H),
3.38-3.52 (m, 1H), 3.63 (d, J=2.40 Hz, 1H), 3.95-4.06 (m, 1H), 4.21
(td, J=9.84, 2.42 Hz, 1H), 4.56 (d, J=7.58 Hz, 1H), 5.58 (br. s.,
1H), 5.66 (d, J=9.44 Hz, 1H), 7.20 (d, J=4.99 Hz, 1H), 8.73 (d,
J=4.99 Hz, 1H), 9.05 (s, 1H).
Synthesis of (+/-)-tert-butyl
(1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-3-methyl-2-(2-(methylsulfonyl)ethox-
y)cyclohexylcarbamate
##STR00188##
[0496] To a solution of (+/-)-tert-butyl
(1R,5S,6S)-5-methyl-6-(2-(methylsulfonyl)ethoxy)-3-(3-nitropyridin-4-yl)c-
yclohex-2-enylcarbamate (1.0 equiv.) in degassed EtOH (0.17 M) was
added Pd/C (0.3 equiv.). The mixture was purged with H.sub.2, and
allowed to stir under H.sub.2 overnight at RT. The reaction was
filtered through a pad of celite and the cake was washed with MeOH.
The organics were concentrated and purified by ISCO SiO.sub.2
chromatography to yield (+/-)-tert-butyl
(1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-3-methyl-2-(2-(methylsulfonyl)ethox-
y)cyclohexylcarbamate in 55% yield. LC/MS (m/z)=428.2 (MH),
R.sub.t=0.59 min. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm
1.07 (d, J=6.85 Hz, 3H), 1.46 (s, 9H), 1.72-1.88 (m, 2H), 2.62 (tt,
J=12.23, 3.30 Hz, 1H), 3.08 (s, 3H), 3.21 (d, J=14.62 Hz, 1H),
3.35-3.47 (m, 1H), 3.58 (br. s., 1H), 3.64 (br. s., 2H), 3.70-3.86
(m, 1H), 3.94-4.10 (m, 1H), 4.10-4.22 (m, 1H), 5.43 (d, J=9.00 Hz,
1H), 6.89 (d, J=5.04 Hz, 1H), 7.98 (d, J=4.99 Hz, 1H), 8.03 (s,
1H).
Synthesis of tert-butyl (1
S,2R,3R,5S)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-y-
l)-3-methyl-2-(2-(methylsulfonyl)ethoxy)cyclohexylcarbamate and
tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(2-(methylsulfonyl)ethoxy)cyclohexylcarbamate
##STR00189##
[0498] EDC (2.0 equiv.) and HOAt (2.0 equiv.) were added to a
solution of (+/-)-tert-butyl
(1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-3-methyl-2-(2-(methylsulfonyl)ethox-
y) cyclohexylcarbamate (1.0 equiv.) and
6-(2,6-difluorophenyl)-5-fluoropicolinic acid (1.5 equiv.) in DMF
(0.08 M). The mixture was stirred at ambient temperature overnight.
The reaction mixture was diluted with water and extracted with
ethyl acetate. The combined extracts were washed sequentially with
1M aqueous sodium hydroxide and brine, dried over sodium sulfate,
filtered, concentrated and purified by ISCO SiO.sub.2
chromatography. Purification was completed via SFC (50% MeOH, 100
mL/min, IC column) to yield tert-butyl
(1S,2R,3R,5S)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(2-(methylsulfonyl)ethoxy) cyclohexylcarbamate (48%
yield, 99% ee) and tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(2-(methylsulfonyl) ethoxy)cyclohexylcarbamate (48%
yield, 99% ee). LC/MS (m/z)=663.2 (MH.sup.+), R.sub.t=0.88 min.
.sup.1H NMR (400 MHz, <cdcl3>) .delta. ppm 0.95 (d, J=6.75
Hz, 3H), 1.45 (s, 8H), 1.50-1.84 (m, 5H), 2.80-2.95 (m, 1H), 3.08
(s, 3H), 3.14-3.29 (m, 1H), 3.32-3.45 (m, 1H), 3.56 (br. s., 1H),
3.63-3.77 (m, 1H), 3.95-4.08 (m, 1H), 4.12 (q, J=7.12 Hz, 1H), 5.36
(d, J=8.56 Hz, 1H), 7.04-7.18 (m, 3H), 7.50 (tt, J=8.47, 6.35 Hz,
1H), 7.77 (t, J=8.56 Hz, 1H), 8.33-8.46 (m, 2H), 9.26 (s, 1H), 9.81
(s, 1H).
Synthesis of (+/-)-tert-butyl
(1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-2-cyano-3-methylcyclohexylcarbamate
##STR00190##
[0500] To a solution of
(+/-)-(1R,2R,4R,6S)-2-(tert-butoxycarbonylamino)-4-(3-(tert-butoxycarbony-
lamino)pyridin-4-yl)-6-methylcyclohexyl methanesulfonate (1.0
equiv.) in DMF (0.20 M) was added NaCN (5.0 equiv.). The solution
was submerged in an 85.degree. C. oil bath and left stirring under
Ar for 16 hrs. The solution was cooled to rt and left stirring
under Ar overnight. The solution was partitioned between EtOAc and
H.sub.2O. The organic layer was washed with Na.sub.2CO.sub.3(sat.),
NaCl.sub.(sat.), dried over MgSO.sub.4, filtered, concentrated. To
a solution of the bis-Boc product (10 equiv.) in DCM (0.20 M) was
added TFA (62.0 equiv.). The mixture was stirred at ambient
temperature for 40 min and concentrated and neutralized with
saturated aqueous sodium bicarbonate. Dioxane (0.15 M) and
Boc.sub.2O (4.0 equiv.) were added. The reaction mixture was
stirred vigorously at ambient temperature for 16 hrs. Volatiles
were removed in vacuo. The aqueous phase was extracted with 10:1
DCM: MeOH. The combined extracts were dried over sodium sulfate,
filtered, and concentrated. The crude material was purified by ISCO
chromatography over silica gel to give (+/-)-tert-butyl
(1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-2-cyano-3-methylcyclohexylcarbamate
in 20% yield. LC/MS (m/z)=331.2 (MH.sup.+), R.sub.t=0.62 min.
Synthesis of tert-butyl
(1R,2S,3S,5R)-2-cyano-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate and tert-butyl
(1S,2R,3R,5S)-2-cyano-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate
##STR00191##
[0502] EDC (1.1 equiv.) and HOAt (1.1 equiv.) were added to a
solution of (+/-)-tert-butyl
(1R,2S,3S,5R)-5-(3-aminopyridin-4-yl)-2-cyano-3-methylcyclohexylcarbamate
(1.0 equiv.) and 6-(2,6-difluorophenyl)-5-fluoropicolinic acid (1.5
equiv.) in DMF (0.11 M). The mixture was stirred at ambient
temperature overnight. The reaction mixture was diluted with water
and extracted with ethyl acetate. The combined extracts were washed
sequentially with 1M aqueous sodium hydroxide and brine, dried over
sodium sulfate, filtered, concentrated and purified by ISCO
SiO.sub.2 chromatography. Purification was completed via SFC (15%
IPA, 100 mL/min, IA column) to yield tert-butyl
(1R,2S,3S,5R)-2-cyano-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate (25% yield, 99% ee) and
tert-butyl
(1S,2R,3R,5S)-2-cyano-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate (27% yield, 99% ee). LC/MS
(m/z)=566.2 (MH.sup.+), R.sub.t=0.90 min.
Synthesis of tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(1H-1,2,3-triazol-1-yl)cyclohexylcarbamate
##STR00192##
[0504] A solution of tert-butyl
(1R,2S,3S,5R)-2-azido-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)p-
yridin-4-yl)-3-methylcyclohexylcarbamate (1.0 equiv.) in vinyl
acetate (0.06 M) to give a suspension was heated in microwave at
160.degree. C. for 1 hr. The reaction was concentrated to yield
tert-butyl
(1R,2S,3S,5R)-5-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-
-yl)-3-methyl-2-(1H-1,2,3-triazol-1-yl)cyclohexylcarbamate in 50%
yield. LC/MS (m/z)=608.3 (MH.sup.+), R.sub.t=0.89 min.
Synthesis of (+/-)-tert-butyl
((1R,2S,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-amino-3-m-
ethylcyclohexyl)carbamate
##STR00193##
[0506] To a degassed a solution of (+/-)-tert-butyl
((1R,2S,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-azido-3-m-
ethylcyclohexyl)carbamate (1.0 equiv.) in ethanol (0.10 M) was
added Pd/C (0.2 equiv.). The mixture was stirred under H.sub.2 for
4 hrs. Filter the mixture over cetlite and wash the cake with MeOH.
Concentrate the filtrate to yield (+/-)-tert-butyl
((1R,2S,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-amino-3-m-
ethylcyclohexyl)carbamate in 88% yield. LC/MS (m/z)=421.3
(MH.sup.+), R.sub.t=0.58 min.
Synthesis of (+/-)-tert-butyl
((1R,2S,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-3-methyl-2--
(methylamino)cyclohexyl)carbamate
##STR00194##
[0508] To a solution of (+/-)-tert-butyl
((1R,2S,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-amino-3-m-
ethylcyclohexyl)carbamate (1.0 equiv.) in MeOH (0.10 M) was added
benzaldehyde (1.3 equiv.). After 3 hrs, sodium cyanotrihydroborate
(2.5 equiv.) was added and the mixture was stirred at rt for 16
hrs. The reaction mixture was quenched by the addition of water,
and volatiles were removed in vacuo. The mixture was extracted with
ethyl acetate. The combined organic phases were dried with sodium
sulfate, filtered and concentrated. The residue was dissolved in
MeOH (0.10 M) and paraformaldehyde (5.0 equiv.) was added. After 16
hrs, sodium cyanotrihydroborate (5.0 equiv.) was added and the
mixture was left stirred at rt for 16 hrs. The reaction was
quenched by the addition of water, and volatiles were removed in
vacuo. The mixture was extracted with DCM. The combined organic
phases were dried with sodium sulfate, filtered, concentrated and
purified by ISCO Chromatography. The product was dissolved in MeOH
(0.10 M) and treated with Pd(OH).sub.2 (0.50 equiv.) under H.sub.2
for 5 hrs at RT. The reaction was filtered through a pad of celite
and the cake was washed with MeOH. The organics were concentrated
to yield (+/-)-tert-butyl
((1R,2S,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-3-methyl-2--
(methylamino)cyclohexyl)carbamate in 75% yield. LC/MS (m/z)=435.2
(MH.sup.+), R.sub.t=0.64 min.
Synthesis of (+/-)-methyl
((1S,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-((tert-butoxycarbonyl)amino)-6-m-
ethylcyclohexyl)(methyl)carbamate
##STR00195##
[0510] To a solution of (+/-)-tert-butyl
((1R,2S,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-3-methyl-2--
(methylamino)cyclohexyl)carbamate (1.0 equiv.) in DCM (0.05 M) at
0.degree. C. was added DIEA (3.0 equiv.) and then methyl
chloroformate (1.5 equiv.). The homogeneous solution was left
standing at 0.degree. C. at for 4 hrs. The reaction was quenched
partitioned between NaHCO.sub.3 solution and EtOAc. The organic
layer was washed with Brine, dried over Na.sub.2SO.sub.4,
concentrated and purified by ISCO chromatography. The product was
treated with 4 M HCl in dioxane (30.0 equiv.) at rt for 1 hour. The
volatiles were removed in vacuo and the solid was pumped on for 5
minutes on the high vac. To the residue was added CH.sub.2Cl.sub.2
(0.15 M), DIEA (5.0 equiv.) and tert-butyl 2,5-dioxopyrrolidin-1-yl
carbonate (1.6 equiv.). The solution was left stirring at rt for 1
hr. The volatiles were removed in vacuo and the residue was
partitioned between EtOAc and H.sub.2O. The organic layer was
washed with Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.), dried over
MgSO.sub.4, filtered, concentrated and purified by ISCO SiO.sub.2
chromatography to yield (+/-)-methyl
((1S,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-((tert-butoxycarbonyl)amino)-6-m-
ethylcyclohexyl)(methyl)carbamate in 20% yield. LC/MS (m/z)=393.2
(MH.sup.+), R.sub.t=0.60 min.
Synthesis of methyl
((1S,2R,4R,6S)-2-((tert-butoxycarbonyl)amino)-4-(3-(6-(2,6-difluorophenyl-
)-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl)(methyl)carbamate
and methyl
((1R,2S,4S,6R)-2-((tert-butoxycarbonyl)amino)-4-(3-(6-(2,6-difluorophenyl-
)-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl)(methyl)carbamate
##STR00196##
[0512] EDC (1.1 equiv.) and HOAt (1.1 equiv.) were added to a
solution of (+/-)-methyl
((1S,2R,4R,6S)-4-(3-aminopyridin-4-yl)-2-((tert-butoxycarbonyl)amino)-6-m-
ethylcyclohexyl)(methyl)carbamate (1.0 equiv.) and
6-(2,6-difluorophenyl)-5-fluoropicolinic acid (1.5 equiv.) in DMF
(0.05 M). The mixture was stirred at ambient temperature overnight.
The reaction mixture was diluted with water and extracted with
ethyl acetate. The combined extracts were washed sequentially with
1M aqueous sodium hydroxide and brine, dried over sodium sulfate,
filtered, concentrated and purified by ISCO SiO.sub.2
chromatography. Purification was completed via SFC (40% EtOH, 100
mL/min, IC column) to yield methyl
((1S,2R,4R,6S)-2-((tert-butoxycarbonyl)amino)-4-(3-(6-(2,6-difluorophenyl-
)-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl)(methyl)carbamate
(15% yield, 99% ee) and methyl
((1R,2S,4S,6R)-2-((tert-butoxycarbonyl)amino)-4-(3-(6-(2,6-difluorophenyl-
)-5-fluoropicolinamido)pyridin-4-yl)-6-methylcyclohexyl)(methyl)carbamate
(15% yield, 99% ee). LC/MS (m/z)=628.3 (MH.sup.+), R.sub.t=0.89
min.
Synthesis of tert-butyl
(3R,4S,5S)-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-5-methyl-4-(1H-1,2,3-triazol-1-yl)piperidin-3-ylcarbamate
##STR00197##
[0514] A solution of tert-butyl
(3R,4S,5S)-4-azido-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyri-
din-4-yl)-5-methylpiperidin-3-ylcarbamate (1.0 equiv.) in vinyl
acetate (0.06 M) to give a suspension was heated at 110.degree. C.
for 88 hrs. The reaction was concentrated and purified by ISCO
chromatography to yield tert-butyl
(3R,4S,5S)-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-5-methyl-4-(1H-1,2,3-triazol-1-yl)piperidin-3-ylcarbamate in 48%
yield. LC/MS (m/z)=609.3 (MH.sup.+), R.sub.t=0.83 min.
Synthesis of tert-butyl
(3R,4S,5S)-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-5-methyl-4-(4-(thiophen-3-yl)-1H-1,2,3-triazol-1-yl)piperidin-3-ylcarbam-
ate
##STR00198##
[0516] In a high pressure vial was added tert-butyl
(3R,4S,5S)-4-azido-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyri-
din-4-yl)-5-methylpiperidin-3-ylcarbamate (1.0 equiv.), copper in
charcoal (0.4 equiv.), 3-ethynyl thiophene (5.0 equiv.) and
triethylamine (1.0 equiv.), in dioxane (0.09 M) to give a black
suspension. The pressure tube was sealed and the mixture was
stirred with heating to 100.degree. C. overnight. The reaction was
cooled to RT, filtered through celite, concentrated to yield
tert-butyl
(3R,4S,5S)-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-5-methyl-4-(4-(thiophen-3-yl)-1H-1,2,3-triazol-1-yl)piperidin-3-ylcarbam-
ate in 48% yield. LC/MS (m/z)=691.2 (MH.sup.+), R.sub.t=0.98
min.
Synthesis of tert-butyl
(3R,4S,5S)-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-5-methyl-4-(4-methyl-1H-1,2,3-triazol-1-yl)piperidin-3-ylcarbamate
##STR00199##
[0518] In a high pressure vial was added tert-butyl
(3R,4S,5S)-4-azido-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyri-
din-4-yl)-5-methylpiperidin-3-ylcarbamate (1.0 equiv.), copper in
charcoal (0.2 equiv.), prop-1-yne (10.0 equiv.) and triethylamine
(1.5 equiv.), in dioxane (0.15 M) to give a black suspension. The
pressure tube was sealed and the mixture was stirred with heating
to 60.degree. C. for 48 hrs. The reaction was cooled to RT,
filtered through celite, concentrated to yield tert-butyl
(3R,4S,5S)-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-5-methyl-4-(4-methyl-1H-1,2,3-triazol-1-yl)piperidin-3-ylcarbamate
in 95% yield. LC/MS (m/z)=623.2 (MH.sup.+), R.sub.t=0.87 min.
Synthesis of tert-butyl
(3R,4S,5S)-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-4-(4-(methoxymethyl)-1H-1,2,3-triazol-1-yl)-5-methylpiperidin-3-ylcarbam-
ate
##STR00200##
[0520] In a high pressure vial was added tert-butyl
(3R,4S,5S)-4-azido-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyri-
din-4-yl)-5-methylpiperidin-3-ylcarbamate (1.0 equiv.), copper in
charcoal (0.2 equiv.), 3-methoxyprop-1-yne (1.5 equiv.) and
triethylamine (1.5 equiv.), in dioxane (0.20 M) to give a black
suspension. The pressure tube was sealed and the mixture was
stirred with heating to 70.degree. C. for 16 hrs. The reaction was
cooled to RT, filtered through celite, concentrated to yield
tert-butyl
(3R,4S,5S)-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyridin-4-yl-
)-4-(4-(methoxymethyl)-1H-1,2,3-triazol-1-yl)-5-methylpiperidin-3-ylcarbam-
ate in 95% yield. LC/MS (m/z)=653.2 (MH.sup.+), R.sub.t=0.86
min.
Synthesis of tert-butyl
((3S,4S,5R)-1-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-4-cyano-5-meth-
ylpiperidin-3-yl)carbamate
##STR00201##
[0522] To a solution of
(3R,4R,5S)-1-(3-(bis(tert-butoxycarbonyl)amino)pyridin-4-yl)-3-(tert-buto-
xycarbonylamino)-5-methylpiperidin-4-ylmethanesulfonate (1.0
equiv.) in DMF (0.10 M) was added NaCN (5.0 equiv.). The mixture
was stirred at 80.degree. C. for 6 hrs and partitioned between
EtOAc and H.sub.2O. The organic layer was washed NaCl.sub.(sat.),
dried over MgSO.sub.4, filtered, concentrated and purified by ISCO
SiO.sub.2 chromatography to yield tert-butyl
((3S,4S,5R)-1-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-4-cyano-5-meth-
ylpiperidin-3-yl)carbamate in 5% yield. LC/MS (m/z)=432.2
(MH.sup.+), R.sub.t=0.73 min.
Synthesis of tert-butyl
(3S,4S,5R)-1-(3-aminopyridin-4-yl)-4-cyano-5-methylpiperidin-3-ylcarbamat-
e
##STR00202##
[0524] Tert-butyl
((3S,4S,5R)-1-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-4-cyano-5-meth-
ylpiperidin-3-yl)carbamate (1.0 equiv.) was treated with 4 M HCl in
dioxane (30.0 equiv.) at rt for 1 hour. The volatiles were removed
in vacuo and the solid was pumped on for 5 minutes on the high vac.
To the residue was added CH2C12 (0.05 M), DIEA (5.0 equiv.) and
Boc-OSu (1.6 equiv.). The solution was left stirring at rt for 1
hr. The volatiles were removed in vacuo and the residue was
partitioned between EtOAc and H2O. The organic layer was washed
with Na2CO3(sat.), NaCl(sat.), dried over MgSO4, filtered,
concentrated to yield tert-butyl
(3S,4S,5R)-1-(3-aminopyridin-4-yl)-4-cyano-5-methylpiperidin-3-ylcarbamat-
e in 100% yield. LC/MS (m/z)=332.1 (MH.sup.+), Rt=0.59 min.
Synthesis of tert-butyl
(4-((3R,4S,5S)-4-azido-3-((tert-butoxycarbonyl)amino)-5-methylpiperidin-1-
-yl)pyridin-3-yl)(tert-butoxycarbonyl)carbamate
##STR00203##
[0526] To a solution of
(3R,4R,5S)-1-(3-(bis(tert-butoxycarbonyl)amino)pyridin-4-yl)-3-((tert-but-
oxycarbonyl)amino)-5-methylpiperidin-4-ylmethanesulfonate (1.0
equiv.) in DMF (0.13 M) was added NaN.sub.3 (5.0 equiv.). The
solution was submerged in an 80.degree. C. oil bath and left
stirring under Ar for 24 hrs. The solution was cooled to rt and
left stirring under Ar overnight. The solution was partitioned
between EtOAc and H.sub.2O. The organic layer was washed with
Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.), dried over MgSO.sub.4,
filtered, concentrated to yield tert-butyl
(4-((3R,4S,5S)-4-azido-3-((tert-butoxycarbonyl)amino)-5-methylpiperidin-1-
-yl)pyridin-3-yl)(tert-butoxycarbonyl)carbamate in 60% yield. LC/MS
(m/z)=548.4 (MH.sup.+), R.sub.t=0.94 min. .sup.1H NMR (400 MHz,
<cdcl3>) .delta. ppm 0.97-1.13 (m, 3H), 1.33-1.52 (m, 30H),
2.03-2.19 (m, 1H), 2.66-2.87 (m, 2H), 3.16 (dd, J=12.72, 2.15 Hz,
1H), 3.22-3.32 (m, 1H), 3.81-4.01 (m, 2H), 4.78 (d, J=9.00 Hz, 1H),
6.76-6.88 (m, 1H), 8.05-8.18 (m, 1H), 8.26-8.37 (m, 1H).
Synthesis of tert-butyl
((3R,4S,5S)-1-(3-aminopyridin-4-yl)-4-azido-5-methylpiperidin-3-yl)carbam-
ate
##STR00204##
[0528] A solution of 4 M HCl in dioxane (30.0 equiv.) was added to
tert-butyl
(4-((3R,4S,5S)-4-azido-3-((tert-butoxycarbonyl)amino)-5-methylpiperidin-1-
-yl)pyridin-3-yl)(tert-butoxycarbonyl)carbamate (1.0 equiv.). The
solution started to go homogeneous for a few minutes, but then a
ppt formed and the solution went very thick. After sitting at rt
for 1 hour, the volatiles were removed in vacuo and the solid was
pumped on for 5 minutes on the high vac. To the residue was added
CH.sub.2Cl.sub.2 (0.11 M), TEA (5.0 equiv.) and Boc.sub.2O (1.0
equiv.). The solution was left stirring at rt for 1 hr. The
volatiles were removed in vacuo and the residue was partitioned
between EtOAc and H.sub.2O. The organic layer was washed with
Na.sub.2CO.sub.3(sat.), NaCl.sub.(sat.), dried over MgSO.sub.4,
filtered, concentrated and purified by ISCO SiO.sub.2
chromatography to yield tert-butyl
((3R,4S,5S)-1-(3-aminopyridin-4-yl)-4-azido-5-methylpiperidin-3-yl)carbam-
ate in 33% yield. LC/MS (m/z)=348.2 (MH.sup.+), R.sub.t=0.70 min.
.sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 1.02-1.18 (m, 3H),
1.36-1.54 (m, 10H), 2.19 (qd, J=6.91, 3.91 Hz, 1H), 2.57 (q,
J=10.96 Hz, 2H), 2.96 (d, J=9.00 Hz, 1H), 3.20 (dd, J=11.15, 3.72
Hz, 1H), 3.55-3.73 (m, 2H), 3.90 (br. s., 1H), 4.01 (br. s., 1H),
4.81 (d, J=8.61 Hz, 1H), 6.72-6.83 (m, 1H), 7.96 (d, J=5.09 Hz,
1H), 8.02 (s, 1H).
Synthesis of tert-butyl
((3R,4S,5S)-4-amino-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyr-
idin-4-yl)-5-methylpiperidin-3-yl)carbamate
##STR00205##
[0530] EDC (1.5 equiv.) and HOAt (1.5 equiv.) were added to a
solution of tert-butyl
((3R,4S,5S)-1-(3-aminopyridin-4-yl)-4-azido-5-methylpiperidin-3-yl)carbam-
ate (1.0 equiv.) and 6-(2,6-difluorophenyl)-5-fluoropicolinic acid
(1.3 equiv.) in DMF (0.20 M). The mixture was stirred at ambient
temperature overnight. The reaction mixture was diluted with water
and extracted with ethyl acetate. The combined extracts were washed
sequentially with 1M aqueous sodium hydroxide and brine, dried over
sodium sulfate, filtered, concentrated and purified by ISCO
SiO.sub.2 chromatography. To a degassed a solution of the azide
(1.0 equiv.) in 2-propanol (0.10 M) was added Pd/C (0.2 equiv.).
The mixture was stirred under H.sub.2 for 48 hrs. Filter the
mixture over cetlite and wash the cake with MeOH. Concentrate the
filtrate to yield tert-butyl
((3R,4S,5S)-4-amino-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyr-
idin-4-yl)-5-methylpiperidin-3-yl)carbamate in 58% yield). LC/MS
(m/z)=557.1 (MH.sup.+), R.sub.t=0.69 min.
Synthesis of
N-(4-((3R,4S,5S)-4-acetamido-3-amino-5-methylpiperidin-1-yl)pyridin-3-yl)-
-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00206##
[0532] Method 4 was followed using tert-butyl
((3R,4S,5S)-4-amino-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyr-
idin-4-yl)-5-methylpiperidin-3-yl)carbamate and acetic anhydride to
give
N-(4-((3R,4S,5S)-4-acetamido-3-amino-5-methylpiperidin-1-yl)pyridin-3-yl)-
-6-(2,6-difluorophenyl)-5-fluoropicolinamidein 40% yield. LC/MS
(m/z)=499.1 (MH.sup.+), R.sub.t=0.58 min.
Synthesis of
N-(4-((3R,4S,5S)-3-amino-5-methyl-4-(methylsulfonamido)piperidin-1-yl)pyr-
idin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide
##STR00207##
[0534] Method 4 was followed using tert-butyl
((3R,4S,5S)-4-amino-1-(3-(6-(2,6-difluorophenyl)-5-fluoropicolinamido)pyr-
idin-4-yl)-5-methylpiperidin-3-yl)carbamate and methanesulfonyl
chloride to give
N-(4-((3R,4S,5S)-3-amino-5-methyl-4-(methylsulfonamido)piperidin--
1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide in
28% yield. LC/MS (m/z)=535.0 (MH.sup.+), R.sub.t=0.58 min.
Synthesis of di-tert-butyl
(4-((3R,4S,5S)-3-((tert-butoxycarbonyl)amino)-5-methyl-4-(methylamino)pip-
eridin-1-yl)pyridin-3-yl)iminodicarbonate
##STR00208##
[0536] To a solution of di-tert-butyl
4-((3R,4S,5S)-4-azido-3-(tert-butoxycarbonylamino)-5-methylpiperidin-1-yl-
)pyridin-3-yliminodicarbonate (1.0 equiv.) in DCM (0.14 M) at rt
was added PMe.sub.3 (2.0 equiv.). After stirring at rt for 2 hr,
PARAFORMALDEHYDE (5.0 equiv.) was added and the mixture was stirred
at rt for another 2.5 hrs. The reaction was added MeOH (0.14 M),
cooled to 0.degree. C. and added NaBH.sub.4 (5.0 equiv.). After 30
min at rt, the reaction was quenched with sat. NaHCO.sub.3 and
extract with EtOAc to yield di-tert-butyl
4-((3R,4S,5S)-3-(tert-butoxycarbonylamino)-5-methyl-4-(methylamino)piperi-
din-1-yl)pyridin-3-yliminodicarbonate in 85% yield. LC/MS
(m/z)=536.3 (MH.sup.+), R.sub.t=0.61 min.
Synthesis of methyl
((3R,4S,5S)-1-(3-aminopyridin-4-yl)-3-((tert-butoxycarbonyl)amino)-5-meth-
ylpiperidin-4-yl)(methyl)carbamate
##STR00209##
[0538] To a solution of di-tert-butyl
4-((3R,4S,5S)-3-(tert-butoxycarbonylamino)-5-methyl-4-(methylamino)piperi-
din-1-yl)pyridin-3-yliminodicarbonate (1.0 equiv.) in DCM (0.10 M)
was added DIEA (3.0 equiv.) the reaction mixture was then cooled to
0.degree. C. To this solution was added methyl chloroformate (1.2
equiv.). The resulting mixture was at RT for 50 min. The reaction
mixture was quenched with NaHCO.sub.3 and diluted with EtOAc. The
aqeuous layer was separated and extracted with EtOAc, the combined
organics were then dried over MgSO.sub.4 and concentrated in
vaccuo. 4 M HCl (43.0 equiv.) in dioxane was added to the residue.
After 1 hr, the volatile was removed in vacuo. To the solution of
the residue in DCM (0.10 M) at 0.degree. C. was added DIEA (3.0
equiv.) and BocOSu (1.0 equiv.). After 60 min at rt, The reaction
mixture was quenched with NaHCO.sub.3 and diluted with EtOAc. The
aqueous layer was separated and extracted with EtOAc, the combined
organics were then dried over MgSO.sub.4 and concentrated in vaccuo
to yield a yellow residue, which was purified by ISCO SiO.sub.2
chromatography to yield methyl
((3R,4S,5S)-1-(3-aminopyridin-4-yl)-3-((tert-butoxycarbonyl)amino)-5-meth-
ylpiperidin-4-yl)(methyl)carbamate in 44% yield. LC/MS (m/z)=394.2
(MH.sup.+), R.sub.t=0.59 min. .sup.1H NMR (400 MHz, <cdcl3>)
.delta. ppm 1.06 (d, J=7.04 Hz, 3H), 1.40-1.52 (m, 10H), 2.28-2.42
(m, 1H), 2.89 (d, J=4.70 Hz, 2H), 3.08 (dd, J=11.93 Hz, 4.50 Hz,
1H), 3.15 (s, 3H), 3.47 (dd, J=11.15, 4.11 Hz, 1H), 3.67-3.79 (m,
5H), 4.13-4.23 (m, 1H), 4.56-4.77 (m, 1H), 6.80 (d, J=5.09 Hz, 1H),
7.97 (d, J=5.48 Hz, 1H), 8.04 (s, 1H).
Synthesis of tert-butyl
((3R,4S,5S)-1-(3-aminopyridin-4-yl)-5-methyl-4-(N-methylacetamido)piperid-
in-3-yl)carbamate
##STR00210##
[0540] To a solution of di-tert-butyl
4-((3R,4S,5S)-3-(tert-butoxycarbonylamino)-5-methyl-4-(methylamino)piperi-
din-1-yl)pyridin-3-yliminodicarbonate (1.0 equiv.) in DCM (0.10 M)
was added DIEA (3.0 equiv.) the reaction mixture was then cooled to
0.degree. C. To this solution was added acetic anhydride (1.2
equiv.). The resulting mixture was at RT for 50 min. The reaction
mixture was quenched with NaHCO.sub.3 and diluted with EtOAc. The
aqeuous layer was separated and extracted with EtOAc, the combined
organics were then dried over MgSO.sub.4 and concentrated in vacuo.
4 M HCl (43.0 equiv.) in Dioxane was added to the residue. After 1
hr, the volatile was removed in vacuo. To the solution of the
residue in DCM (0.10 M) at 0.degree. C. was added DIEA (3.0 equiv.)
and BocOSu (1.0 equiv.). After 60 min at rt, The reaction mixture
was quenched with NaHCO.sub.3 and diluted with EtOAc. The aqeuous
layer was separated and extracted with EtOAc, the combined organics
were then dried over MgSO.sub.4 and concentrated in vaccuo to yield
a yellow residue, which was purified by SiO.sub.2 chromatography to
yield tert-butyl
((3R,4S,5S)-1-(3-aminopyridin-4-yl)-5-methyl-4-(N-methylacetamido)piperid-
in-3-yl)carbamate in 60% yield. LC/MS (m/z)=378.2 (WO, R.sub.t=0.50
min. .sup.1HNMR (400 MHz, <cdcl3>) .delta. ppm 1.06 (d,
J=7.83 Hz, 3H), 1.39-1.50 (m, 9H), 2.20 (br. s., 3H), 2.34-2.48 (m,
1H), 2.84-3.28 (m, 6H), 3.77 (d, J=18.00 Hz, 2H), 4.19-4.62 (m,
1H), 6.82 (d, J=5.09 Hz, 1H), 7.97 (br. s., 1H), 8.05 (br. s.,
1H).
Synthesis of tert-butyl
tert-butoxycarbonyl(4-((3R,4S,5S)-3-((tert-butoxycarbonyl)amino)-4-(4-(hy-
droxymethyl)-1H-1,2,3-triazol-1-yl)-5-methylpiperidin-1-yl)pyridin-3-yl)ca-
rbamate
##STR00211##
[0542] In a round-bottom flask was added tert-butyl
(4-((3R,4S,5S)-4-azido-3-((tert-butoxycarbonyl)amino)-5-methylpiperidin-1-
-yl)pyridin-3-yl)(tert-butoxycarbonyl) carbamate (1.0 equiv.),
copper in charcoal (0.02 equiv.) and propargyl alcohol (1.0 equiv.)
in t-BuOH/H.sub.2O (0.15 M) to give a blue solution. And sodium
ascorbate (0.1 equiv.) was added. The mixture was stirred at rt for
16 hrs. The reaction was diluted with H.sub.2O and cooled to
0.degree. C., then filtered, and the precipitate was collected to
yield tert-butyl
tert-butoxycarbonyl(4-((3R,4S,5S)-3-((tert-butoxycarbonyl)amino)-4-(4-(hy-
droxymethyl)-1H-1,2,3-triazol-1-yl)-5-methylpiperidin-1-yl)pyridin-3-yl)ca-
rbamate in 85% yield. LC/MS (m/z)=604.3 (MH.sup.+), R.sub.t=0.69
min.
Synthesis of tert-butyl
(3R,4S,5S)-1-(3-aminopyridin-4-yl)-4-(4-(hydroxymethyl)-1H-1,2,3-triazol--
1-yl)-5-methylpiperidin-3-ylcarbamate
##STR00212##
[0544] To di-tert-butyl
4-((3R,4S,5S)-3-(tert-butoxycarbonylamino)-4-(4-(hydroxymethyl)-1H-1,2,3--
triazol-1-yl)-5-methylpiperidin-1-yl)pyridin-3-yliminodicarbonate
(1.0 equiv.) was added 4 M HCl (30.0 equiv.) in dioxane. After 1
hr, the volatile was removed in vacuo. To the solution of the
residue in DCM (0.10 M) at 0.degree. C. was added DIEA (30.0
equiv.) and BocOSu (1.0 equiv.). After 4 hrs at rt, the reaction
mixture was quenched with NaHCO.sub.3 and diluted with EtOAc. The
aqeuous layer was separated and extracted with EtOAc, the combined
organics were then dried over MgSO.sub.4 and concentrated in vacuo
to yield a yellow residue, which was purified by ISCO SiO.sub.2
chromatography to yield tert-butyl
(3R,4S,5S)-1-(3-aminopyridin-4-yl)-4-(4-(hydroxymethyl)-1H-1,2,3-triazol--
1-yl)-5-methylpiperidin-3-ylcarbamate in 57% yield. LC/MS
(m/z)=404.3 (MH.sup.+), R.sub.t=0.47 min.
Synthesis of
N-(4-((3R,4S,5S)-3-amino-4-(4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl)-5-me-
thylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinami-
de
##STR00213##
[0546] To a round-bottomed flask was added tert-butyl
(3R,4S,5S)-1-(3-aminopyridin-4-yl)-4-(4-(hydroxymethyl)-1H-1,2,3-triazol--
1-yl)-5-methylpiperidin-3-ylcarbamate (1.0 equiv.) and BSA (2.5
equiv.) in Acetonitrile/DMF (4/1, 0.05 M) to give a orange
suspension. The mixture was stirred at room temperature for 1 hr at
which time all of the solids had dissolved into solution. The
mixture was taken to dryness. The crude in DMF (0.10 M) was added
EDCI (2.4 equiv.), HOAT (2.4 equiv.) and
6-(2,6-difluorophenyl)-5-fluoropicolinic acid (2.4 equiv.). The
mixture was stirred at rt for 16 hrs. The reaction was taken to
dryness and dissolved in EtOAc and cooled to 0.degree. C. to
precipitate the urea byproduct. The mixture was filtered and the
organics were washed with H.sub.2O, dried over MgSO.sub.4, filtered
and concentrated. To a solution of the residue in EtOH (0.01 M) was
added potassium carbonate (10.0 equiv.). The mixture was stirred at
room temperature for 2 hrs. The reaction was diluted with DCM and
quenched with NaHCO.sub.3(aq.). The organics were separated and the
aqueous solution was extracted with DCM. The combined organics were
dried over MgSO.sub.4, filtered, and concentrated. The crude
alcohol was then dissolved in 25% TFA/DCM. The solution was stirred
at room temp for 1 hr. The volatiles were removed on the rotovap.
The crude product was taken up in DMSO, filtered, purified via
prep-HPLC to yield
N-(4-((3R,4S,5S)-3-amino-4-(4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl)-5-me-
thylpiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinami-
de in 69% yield. LC/MS (m/z)=539.3 (MH.sup.+), R.sub.t=0.57
min.
Synthesis of (+/-)-tert-butyl
((1R,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-3-methyl-2-oxo-
cyclohexyl)carbamate
##STR00214##
[0548] To a solution of (+/-)-tert-butyl
((1R,2R,3S,5R)-5-(3-((tert-butoxycarbonyl)amino)
pyridin-4-yl)-2-hydroxy-3-methylcyclohexyl)carbamate (1.0 equiv.)
DCM (0.10 M) was added Dess-Martin Periodinane (1.2 equiv.). The
mixture was stirred at room temperature for 18 hrs. The volatiles
were removed under reduced pressure. The residue was partitioned
between EtOAc and 10% sodium thiosulfate/sat. sodium bicarbonate
aqueous solution. The organic was washed with brine, dried over
sodium sulfate, filtered, concentrated and purified by ISCO
chromatography to afford (+/-)-tert-butyl
((1R,3S,5R)-5-(3-acetamidopyridin-4-yl)-3-methyl-2-oxocyclohexyl)
carbamate in 65% yield. LC/MS (m/z)=420.2 (MH.sup.+), R.sub.t=0.73
min.
Synthesis of (+/-)-benzyl
((1R,3S,5R)-5-(3-((benzoxycarbonyl)amino)pyridin-4-yl)-3-methyl-2-oxocycl-
ohexyl)carbamate
##STR00215##
[0550] (+/-)-Tert-butyl
((1R,3S,5R)-5-(3-acetamidopyridin-4-yl)-3-methyl-2-oxocyclo
hexyl)carbamate (1.0 equiv.) was dissolved in 4.0 M HCl (50.0
equiv.) in p-dioxane. After stirring at room temperature for 16
hrs, the mixture was concentrated. The crude was dissolved in DCM
(0.20 M) and CBZ-OSu (5.0 equiv.) was added followed by DIEA (8.0
equiv.). The reaction was stirred at room temperature for 20 hrs.
The solution was diluted with EtOAc and washed with water,
sat.sodium bicarbonate, and brine and dried over sodium sulfate,
filtered and concentrated. The crude residue was purified by ISCO
chromatography to afford (+/-)-benzyl
((1R,3S,5R)-5-(3-((benzoxycarbonyl)amino)
pyridin-4-yl)-3-methyl-2-oxocyclohexyl)carbamate in 50% yield.
LC/MS (m/z)=488.2 (MH.sup.+), R.sub.t=0.79 min.
Synthesis of (+/-)-benzyl
((6R,8R,10S)-8-(3-((benzoxycarbonyl)amino)pyridin-4-yl)-10-methyl-1,4-dio-
xaspiro[4.5]decan-6-yl)carbamate
##STR00216##
[0552] (+/-)-Benzyl ((1R,3S,5R)-5-(3-((benzoxycarbonyl)amino)
pyridin-4-yl)-3-methyl-2-oxocyclohexyl)carbamate (1.0 equiv.) was
dissolved in dry THF (0.20 M) under nitrogen and ETHYLENE GLYCOL
(8.0 equiv.) was added followed by BF.sub.3.OEt.sub.2 (1.4 equiv.).
The solution was microwave vial and heated at 100.degree. C. for 60
mins. The solution was diluted with EtOAc and washed with sat.
sodium bicarbonate, water, brine, dried over sodium sulfate and
concentrated. The crude material was purified by ISCO
chromatography to afford (+/-)-benzyl
((6R,8R,10S)-8-(3-((benzoxycarbonyl)
amino)pyridin-4-yl)-10-methyl-1,4-dioxaspiro[4.5]decan-6-yl)carbamate
in 100% yield. LC/MS (m/z)=532.2 (MH.sup.+), R.sub.t=0.81 min.
Synthesis of benzyl
(6S,8S,10R)-8-(3-aminopyridin-4-yl)-10-methyl-1,4-dioxaspiro[4.5]decan-6--
ylcarbamate and benzyl
((6R,8R,10S)-8-(3-aminopyridin-4-yl)-10-methyl-1,4-dioxaspiro[4.5]decan-6-
-yl)carbamate
##STR00217##
[0554] (+/-)-Benzyl
((6R,8R,10S)-8-(3-((benzoxycarbonyl)amino)pyridin-4-yl)-10-methyl-1,4-dio-
xaspiro[4.5]decan-6-yl)carbamate (1.0 equiv.) was dissolved in MeOH
(0.10 M) and degassed with Argon to vacuum. Pd/C (0.05 equiv.) was
added and the mixture was stirred under a H.sub.2 balloon for 20
hrs. The mixture was filtered and concentrated. The crude and
CBZ-OSu (0.99 equiv.) were dissolved in DCM (0.20 M). The reaction
was stirred at room temperature for 2 hrs. The solution was diluted
with EtOAc and washed with water, dried over sodium sulfate,
filtered and concentrated. The crude residue was purified by ISCO
chromatography. Purification was completed via SFC (40% EtOH, 15
mL/min, OJ column) to yield benzyl
(6S,8S,10R)-8-(3-aminopyridin-4-yl)-10-methyl-1,4-dioxaspiro[4.5]decan-6--
ylcarbamate (28% yield, 99% ee) and benzyl
(6R,8R,10S)-8-(3-aminopyridin-4-yl)-10-methyl-1,4-dioxaspiro[4.5]decan-6--
ylcarbamate (13% yield, 99% ee). LC/MS (m/z)=398.2 (MH.sup.+),
R.sub.t=0.64 min.
Method 7
[0555] A solution of N-Boc protected amine was treated with excess
4M HCl/dioxane for 14 hours or with 25% TFA/CH.sub.2Cl.sub.2 for 2
hours. Upon removal of the volatiles in vacuo, the material was
purified by RP HPLC yielding after lyophilization the amide product
as the TFA salt. Alternatively, the HPLC fractions could be added
to EtOAc and solid Na.sub.2CO.sub.3, separated and washed with
NaCl.sub.(sat.). Upon drying over MgSO.sub.4, filtering and
removing the volatiles in vacuo the free base was obtained. Upon
dissolving in MeCN/H.sub.2O, adding 1 eq. of 1 N HCl and
lyophilizing, the HCl salt of the amide product was obtained.
[0556] If an OBn or NCbz group was present, it was deprotected by
treatment with 10% Pd/C (0.2 equiv.) under an atmosphere of
hydrogen in ethyl acetate and methanol (1:2). Upon completion, the
reaction was filtered through Celite, washed with methanol, and the
filtrate was concentrated in vacuo.
[0557] If a CO.sub.2Me group was present, it could be converted to
the corresponding CO.sub.2H following Method 2.
[0558] Following the procedures of Method 7, the following
compounds were prepared:
TABLE-US-00002 TABLE 1 LC/MS (MH+ LC/MS Ex on (Rf on # Structure
UPLC) UPLC) Chemical Name 1 ##STR00218## 508.2 0.60
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-(1H-1,2,3- triazol-1-
yl)cyclohexyl)pyridin-3-yl)-6- (2,6-difluorophenyl)-5-
fluoropicolinamide 2 ##STR00219## 528.2 0.62 N-(4-((1R,3R,4S,5S)-3-
amino-4-(2- methoxyacetamido)-5- methylcyclohexyl)pyridin-3-
yl)-6-(2,6-difluorophenyl)-5- fluoropicolinamide 3 ##STR00220##
526.3 0.67 N-(4-((1R,3R,4S,5S)-3- amino-4-isobutyramido-5-
methylcyclohexyl)pyridin-3- yl)-6-(2,6-difluorophenyl)-5-
fluoropicolinamide 4 ##STR00221## 591.2 0.72
N-(4-((3R,4S,5S)-3-amino-5- methyl-4-(4-(thiophen-3-yl)-
1H-1,2,3-triazol-1- yl)piperidin-1-yl)pyridin-3-yl)-
6-(2,6-difluorophenyl)-5- fluoropicolinamide 5 ##STR00222## 509.2
0.57 N-(4-((3R,4S,5S)-3-amino- 5-methyl-4-(1H-1,2,3-
triazol-1-yl)piperidin-1- yl)pyridin-3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 6 ##STR00223## 470.3 0.61
N-(4-((1R,3R,5S,Z)-3- amino-4-(hydroxyimino)-5-
methylcyclohexyl)pyridin- 3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 7 ##STR00224## 523.1 0.60
N-(4-((3R,4S,5S)-3-amino-5- methyl-4-(4-methyl-1H-1,2,3-
triazol-1-yl)piperidin-1- yl)pyridin-3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 8 ##STR00225## 528.2 0.65
ethyl (1S,2R,4R,6S)-2- amino-4-(3-(6-(2,6- difluorophenyl)-5-
fluoropicolinamido) pyridin-4-yl)-6- methylcyclohexyl- carbamate 9
##STR00226## 542.3 0.68 isopropyl (1S,2R,4R,6S)-2-
amino-4-(3-(6-(2,6- difluorophenyl)-5- fluoropicolinamido)
pyridin-4-yl)-6- methylcyclohexyl- carbamate 10 ##STR00227## 512.1
0.62 N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-
propionamidocyclohexyl) pyridin-3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 11 ##STR00228## 455.1 0.57 N-(4-((1R,3R,5S)-3-
amino-5-methyl-4- oxocyclohexyl)pyridin-3-
yl)-6-(2,6-difluorophenyl)- 5-fluoropicolinamide 12 ##STR00229##
484.2 0.63 N-(4-((1R,3R,5S,E)-3- amino-4-(methoxyimino)-
5-methylcyclohexyl) pyridin-3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 13 ##STR00230## 484.2 0.64 N-(4-((1R,3R,5S,Z)-3-
amino-4-(methoxyimino)- 5-methylcyclohexyl) pyridin-3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 14 ##STR00231## 553.2 0.60
N-(4-((3R,4S,5S)-3-amino-4- (4-(methoxymethyl)-1H-
1,2,3-triazol-1-yl)-5- methylpiperidin-1-yl)pyridin-
3-yl)-6-(2,6-difluorophenyl)-5- fluoropicolinamide 15 ##STR00232##
549.2 0.58 N-(4-((1R,3R,4S,5S)-3- amino-4-(2-
hydroxyethylsulfonyl)-5- methylcyclohexyl)pyridin- 3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 16 ##STR00233## 534.2 0.57
N-(4-((1S,3S,4S,5R)-3- amino-5-methyl-4- (methylsulfonamido)
cyclohexyl)pyridin-3- yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 17 ##STR00234## 534.2 0.57
N-(4-((1R,3R,4R,5S)-3- amino-5-methyl-4- (methylsulfonamido)
cyclohexyl)pyridin-3- yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 18 ##STR00235## 457.0 0.57 N-(4-((3R,4S,5S)-3,4-
diamino-5-methylpiperidin- 1-yl)pyridin-3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 19 ##STR00236## 535.0 0.58
N-(4-((3R,4S,5S)- 3-amino-5-methyl-4- (methylsulfonamido)
piperidin-1-yl) pyridin-3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 20 ##STR00237## 563.3 0.61
N-(4-((1R,3R,4S,5S)-3- amino-4-((2- methoxyethyl)sulfonyl)-5-
methylcyclohexyl)pyridin-3- yl)-6-(2,6-difluorophenyl)-5-
fluoropicolinamide 21 ##STR00238## 499.1 0.58 N-(4-((3R,4S,5S)-4-
acetamido-3-amino-5- methylpiperidin-1- yl)pyridin-3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 22 ##STR00239## 514.2 0.61
methyl (1S,2R,4R,6S)-2- amino-4-(3-(6-(2,6- difluorophenyl)-5-
fluoropicolinamido) pyridin-4-yl)-6- methylcyclohexyl- carbamate 23
##STR00240## 514.3 0.61 methyl (1R,2S,4S,6R)-2- amino-4-(3-(6-(2,6-
difluorophenyl)-5- fluoropicolinamido) pyridin-4-yl)-6-
methylcyclohexyl- carbamate 24 ##STR00241## 498.2 0.59
N-(4-((1S,3S,4R,5R)-4- acetamido-3-amino-5-
methylcyclohexyl)pyridin- 3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 25 ##STR00242## 498.3 0.59
N-(4-((1R,3R,4S,5S)-4- acetamido-3-amino-5-
methylcyclohexyl)pyridin- 3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 26 ##STR00243## 534.2 0.58
N-(4-((1S,3S,4R,5R)-3- amino-5-methyl-4- (methylsulfonamido)
cyclohexyl)pyridin-3- yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 27 ##STR00244## 534.2 0.58
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4- (methylsulfonamido)
cyclohexyl)pyridin-3- yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 28 ##STR00245## 482.1 0.64
N-(4-((1S,3S,4R,5R)-3- amino-4-azido-5- methylcyclohexyl)pyridin-3-
yl)-6-(2,6-difluorophenyl)-5- fluoropicolinamide 29 ##STR00246##
482.2 0.63 N-(4-((1R,3R,4S,5S)-3- amino-4-azido-5-
methylcyclohexyl)pyridin-3- yl)-6-(2,6-difluorophenyl)-5-
fluoropicolinamide 30 ##STR00247## 498.3 0.57
N-(4-((1R,3R,4R,5S)-4- acetamido-3-amino-5-
methylcyclohexyl)pyridin-3- yl)-6-(2,6-difluorophenyl)-5-
fluoropicolinamide 31 ##STR00248## 503.2 0.59
N-(4-((1R,3R,4R,5S)-3- amino-5-methyl-4-((R)-
methylsulfinyl)cyclohexyl) pyridin-3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 32 ##STR00249## 503.2 0.57
N-(4-((1R,3R,4R,5S)-3- amino-5-methyl-4-((S)-
methylsulfinyl)cyclohexyl) pyridifluorophenyl)-5-
fluoropicolinamide 33 ##STR00250## 487.2 0.64
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4- (methylthio)cyclohexyl)
pyridin-3-yl)-6- (2,6-difluorophenyl)- 5-fluoropicolinamide 34
##STR00251## 519.2 0.60 N-(4-((1R,3R,4R,5S)-3- amino-5-methyl-4-
(methylsulfonyl)cyclohexyl) pyridin-3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 35 ##STR00252## 487.2 0.65
N-(4-((1R,3R,4R,5S)-3- amino-5-methyl-4- (methylthio)cyclohexyl)
pyridin-3-yl)-6- (2,6-difluorophenyl)- 5-fluoropicolinamide 36
##STR00253## 503.2 0.57 N-(4-((1R,3R,4S,5S)-3-
amino-5-methyl-4-((R)- methylsulfinyl)cyclohexyl)
pyridin-3-yl)-6-(2,6- difluorophenyl)-5- fluoropicolinamide 37
##STR00254## 519.2 0.58 N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-
(methylsulfonyl)cyclohexyl) pyridin-3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 38 ##STR00255## 533.3 0.60
(1R,2R,4R,6S)-2-amino-4-(3- (6-(2,6-difluorophenyl)-5-
fluoropicolinamido)pyridin-4- yl)-6-methylcyclohexyl
dimethylphosphinate 39 ##STR00256## 533.3 0.59
(1S,2R,4R,6S)-2-amino-4-(3- (6-(2,6-difluorophenyl)-5-
fluoropicolinamido)pyridin-4- yl)-6-methylcyclohexyl
dimethylphosphinate 40 ##STR00257## 535.2 0.60
(1S,2R,4R,6S)-2-amino-4-(3- (6-(2,6-difluorophenyl)-5-
fluoropicolinamido)pyridin-4- yl)-6-methylcyclohexyl
methanesulfonate 41 ##STR00258## 535.2 0.61
(1R,2R,4R,6S)-2-amino-4-(3- (6-(2,6-difluorophenyl)-5-
fluoropicolinamido)pyridin-4- yl)-6-methylcyclohexyl
methanesulfonate
Method 8
[0559] A homogeneous solution of 1 eq each of amine, carboxylic
acid, HOAT and EDC in DMF, at a concentration of 0.5 M, was left
standing for 24 hours at which time water and ethyl acetate were
added. The organic phase was dried with sodium sulfate and purified
via silica gel column chromatography eluting with ethyl acetate and
hexanes to give the desired protected amide product. Alternatively
the crude reaction mixture was directly purified by HPLC. Upon
lyophilization, the TFA salt of the protected amide product was
obtained. Alternatively, the HPLC fractions could be added to EtOAc
and solid Na.sub.2CO.sub.3, separated and washed with
NaCl.sub.(sat.). Upon drying over MgSO.sub.4, filtering and
removing the volatiles in vacuo, the protected amide product was
obtained as a free base. Alternatively, the crude reaction mixture
was used for the deprotection step without further
purification.
[0560] If an N-Boc protected amine was present, it was removed by
treating with excess 4M HCl/dioxane for 14 hours or by treating
with 25% TFA/CH.sub.2Cl.sub.2 for 2 hours. Upon removal of the
volatiles in vacuo, the material was purified by RP HPLC yielding
after lyophilization the amide product as the TFA salt.
Alternatively, the HPLC fractions could be added to EtOAc and solid
Na.sub.2CO.sub.3, separated and washed with NaCl.sub.(sat.). Upon
drying over MgSO.sub.4, filtering and removing the volatiles in
vacuo the free base was obtained. Upon dissolving in MeCN/H.sub.2O,
adding 1 eq. of 1 N HCl and lyophilizing, the HCl salt of the amide
product was obtained.
[0561] If an N-Boc, OAc group were present, prior to Boc
deprotection, the acetate group could be cleaved by treating with
K.sub.2CO.sub.3 (2.0 equiv.) in ethanol at a concentration of 0.1 M
for 24 hours.
[0562] If a TBDMS ether was present, it was deprotected prior to
Boc removal by treating with 6N HCl, THF, methanol (1:2:1) at room
temperature for 12 h. After removal of volatiles in vacuo, the Boc
amino group was deprotected as described above. Alternatively, the
TBDMS ether and Boc group could be both deprotected with 6N HCl,
THF, methanol (1:2:1) if left at rt for 24 hours, or heated at
60.degree. C. for 3 hours.
[0563] If a OBn or Cbz protecting group was present, it was
deprotected by treatment with 10% Pd/C (0.2 equiv.) under an
atmosphere of hydrogen in ethyl acetate and methanol (1:2). Upon
completion, the reaction was filtered through Celite, washed with
methanol, and the filtrate was concentrated in vacuo.
[0564] If a CO.sub.2Me group was present, it could be converted to
the corresponding CO.sub.2H following Method 2.
[0565] Following the procedures of Method 8, the following
compounds were prepared:
TABLE-US-00003 TABLE 2 LC/MS (MH+ LC/MS Ex on (Rf on # Structure
UPLC) UPLC) Chemical Name 42 ##STR00259## 499.2 0.64
N-(4-((6R,8R,10S)-6-amino- 10-methyl-1,4- dioxaspiro[4.5]decan-8-
yl)pyridin-3-yl)-6-(2,6- difluorophenyl)-5- fluoropicolinamide 43
##STR00260## 499.2 0.64 N-(4-((6S,8S,10R)-6-amino- 10-methyl-1,4-
dioxaspiro[4.5]decan-8- yl)pyridin-3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 44 ##STR00261## 529.3 0.63 3-((1R,2R,4R,6S)-
2-amino-4- (3-(6-(2,6- difluorophenyl)-5- fluoropicolinamido)
pyridin-4-yl)-6- methylcyclohexyloxy) propanoic acid 45
##STR00262## 529.3 0.62 3-((1S,2S,4S,6R)- 2-amino-4- (3-(6-(2,6-
difluorophenyl)-5- fluoropicolinamido) pyridin-4-yl)-6-
methylcyclohexyloxy) propanoic acid 46 ##STR00263## 531.3 0.59
methyl (1R,2S,4S,6R)- 2-amino-4-(3-(5- amino-2-(2,6-
difluorophenyl) thiazole-4- carboxamido) pyridin-4-yl)-6-
methylcyclohexyl (methyl)carbamate 47 ##STR00264## 531.3 0.58
methyl (1S,2R,4R,6S)- 2-amino-4-(3-(5- amino-2-(2,6-
difluorophenyl) thiazole-4- carboxamido) pyridin-4-yl)-6-
methylcyclohexyl (methyl)carbamate 48 ##STR00265## 508.3 0.59
N-(4-((1S,3S,4R,5R)- 3-amino-5-methyl- 4-(1H-1,2,4-
triazol-1-yl)cyclohexyl) pyridin-3-yl)-6- (2,6-difluorophenyl)-5-
fluoropicolinamide 49 ##STR00266## 508.3 0.59 N-(4-((1R,3R,4S,5S)-
3-amino-5-methyl- 4-(1H-1,2,4- triazol-1-yl) cyclohexyl)pyridin-
3-yl)-6- (2,6-difluorophenyl)-5- fluoropicolinamide 50 ##STR00267##
459.2 0.58 N-(4-((1R,3R,4S,5S)-3- amino-4-methoxy-5-
methylcyclohexyl) pyridin-3-yl)-2-(2,6- difluorophenyl)thiazole-
4-carboxamide 51 ##STR00268## 454.3 0.54 N-(4-((1R,3R,4S,5S)-3-
amino-4-methoxy-5- methylcyclohexyl) pyridin-3-yl)-2-(2,6-
difluorophenyl)pyrimidine- 4-carboxamide 52 ##STR00269## 542.2 0.68
methyl (1R,2S,4S,6R)-2- amino-4-(3-(6- (2,6-difluoro-4-
methylphenyl)-5- fluoropicolinamido) pyridin-4-yl)-6-
methylcyclohexyl (methyl)carbamate 53 ##STR00270## 542.2 0.67
methyl (1S,2R,4R,6S)- 2-amino-4-(3-(6- (2,6-difluoro-4-
methylphenyl)-5- fluoropicolinamido) pyridin-4-yl)-6-
methylcyclohexyl (methyl)carbamate 54 ##STR00271## 470.0 0.57
5-amino-N-(4- ((3S,4S,5R)-3- amino-4-cyano-5- methylpiperidin-1-yl)
pyridin-3-yl)-2-(2,6- difluorophenyl) thiazole-4- carboxamide 55
##STR00272## 481.1 0.66 N-(4-((3S,4S,5R)- 3-amino-4-cyano-5-
methylpiperidin-1- yl)pyridin-3-yl)-6-(2,6-
difluoro-4-methylphenyl)- 5-fluoropicolinamide 56 ##STR00273##
509.2 0.57 N-(4-((3R,4S,5S)- 3-amino-5-methyl-
4-(1H-1,2,4-triazol-1- yl)piperidin-1-yl) pyridin-3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 57 ##STR00274## 467.0 0.60
N-(4-((3S,4S,5R)- 3-amino-4-cyano-5- methylpiperidin-1-
yl)pyridin-3-yl)-6-(2,6- difluorophenyl)-5- fluoropicolinamide 58
##STR00275## 543.2 0.67 methyl (3R,4S,5S)-3-amino-
1-(3-(6-(2,6-difluoro-4- methylphenyl)-5- fluoropicolinamido)
pyridin-4-yl)-5- methylpiperidin-4- yl(methyl)carbamate 59
##STR00276## 528.1 0.64 methyl (1R,2S,4S,6R)-2- amino-4-(3-(6-(2,6-
difluorophenyl)-5- fluoropicolinamido) pyridin-4-yl)-6-
methylcyclohexyl(methyl) carbamate 60 ##STR00277## 528.2 0.63
methyl (1S,2R,4R,6S)-2- amino-4-(3-(6-(2,6- difluorophenyl)-5-
fluoropicolinamido) pyridin-4-yl)-6- methylcyclohexyl(methyl)
carbamate 61 ##STR00278## 529.2 0.63 methyl (3R,4S,5S)-3-amino-
1-(3-(6-(2,6-difluorophenyl)- 5-fluoropicolinamido)pyridin-
4-yl)-5-methylpiperidin-4- yl(methyl)carbamate 62 ##STR00279##
485.1 0.69 N-(4-((1R,3R,4R,5S)-3- amino-4-methoxy-5-
methylcyclohexyl)pyridin-3- yl)-6-(2,6-difluoro-4- methylphenyl)-5-
fluoropicolinamide 63 ##STR00280## 513.2 0.61 5-amino-N-(4-
((1S,3S,4R,5R)-3-amino-4- (2-cyanoethoxy)-5-
methylcyclohexyl)pyridin-3- yl)-2-(2,6- difluorophenyl)thiazole-4-
carboxamide 64 ##STR00281## 513.2 0.61 5-amino-N-(4-
((1R,3R,4S,5S)-3-amino-4- (2-cyanoethoxy)-5-
methylcyclohexyl)pyridin-3- yl)-2-(2,6- difluorophenyl)thiazole-4-
carboxamide 65 ##STR00282## 513.2 0.53 N-(4-((3R,4S,5S)-3-
amino-5-methyl-4-(N- methylacetamido)piperidin-
1-yl)pyridin-3-yl)-6-(2,6- difluorophenyl)-5- fluoropicolinamide 66
##STR00283## 539.3 0.57 N-(4-((3R,4S,5S)-3-amino-4-
(4-(hydroxymethyl)-1H-1,2,3- triazol-1-yl)-5-
methylpiperidin-1-yl)pyridin- 3-yl)-6-(2,6-difluorophenyl)-5-
fluoropicolinamide 67 ##STR00284## 466.1 0.61
N-(4-((1S,3S,4R,5R)-3- amino-4-cyano-5- methylcyclohexyl)pyridin-3-
yl)-6-(2,6-difluorophenyl)-5- fluoropicolinamide 68 ##STR00285##
466.1 0.61 N-(4-((1R,3R,4S,5S)-3- amino-4-cyano-5-
methylcyclohexyl)pyridin-3- yl)-6-(2,6-difluorophenyl)-5-
fluoropicolinamide 69 ##STR00286## 474.1 0.58 5-amino-N-(4-
((1R,3R,4R,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-2-(2,6- difluorophenyl)thiazole- 4-carboxamide 70
##STR00287## 469.1 0.54 5-amino-N-(4- ((1R,3R,4R,5S)-3-
amino-4-methoxy-5- methylcyclohexyl) pyridin-3-yl)-2-(2,6-
difluorophenyl)pyrimidine- 4-carboxamide 71 ##STR00288## 488.2 0.68
5-amino-N-(4- ((1R,3R,4S,5S)-3- amino-4-ethoxy-5- methylcyclohexyl)
pyridin-3-yl)-2-(2,6- difluorophenyl)thiazole- 4-carboxamide 72
##STR00289## 518.1 0.60 methyl ((3R,4S,5S)- 3-amino-1-(2-(5-
amino-2-(2,6- difluorophenyl)thiazole- 4-carboxamido)phenyl)-
5-methylpiperidin-4- yl)carbamate 73 ##STR00290## 577.2 0.65
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-(2- (methylsulfonyl)ethoxy)
cyclohexyl)pyridin- 3-yl)-6-(2,6-difluoro- 4-methylphenyl)-5-
fluoropicolinamide 74 ##STR00291## 577.2 0.65
N-(4-((1S,3S,4R,5R)-3- amino-5-methyl-4-(2- (methylsulfonyl)ethoxy)
cyclohexyl)pyridin-3- yl)-6-(2,6-difluoro-4- methylphenyl)-5-
fluoropicolinamide 75 ##STR00292## 471.1 0.66
N-(4-((1R,3R,4R,5S)-3- amino-4-methoxy-5-
methylcyclohexyl)pyridin-3- yl)-6-(2,6-difluorophenyl)-
5-fluoropicolinamide 76 ##STR00293## 471.1 0.67
N-(4-((1S,3S,4S,5R)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluorophenyl)-5- fluoropicolinamide 77
##STR00294## 563.3 0.61 N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-(2-
(methylsulfonyl)ethoxy) cyclohexyl)pyridin- 3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 78 ##STR00295## 563.3 0.61
N-(4-((1S,3S,4R,5R)-3- amino-5-methyl-4-(2- (methylsulfonyl)ethoxy)
cyclohexyl)pyridin- 3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 79 ##STR00296## 469.3 0.53 5-amino-N-(4-
((1R,3R,4S,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-2-(2,6- difluorophenyl) pyrimidine-4- carboxamide 80
##STR00297## 529.1 0.69 methyl ((3R,4S,5S)-3-amino-
1-(3-(6-(2,6-difluoro-4- methylphenyl)-5- fluoropicolinamido)
pyridin-4-yl)-5- methylpiperidin-4- yl)carbamate 81 ##STR00298##
515.1 0.64 methyl ((3R,4S,5S)- 3-amino-1-(3-(6-
(2,6-difluorophenyl)- 5-fluoropicolinamido) pyridin-4-yl)-5-
methylpiperidin-4- yl)carbamate 82 ##STR00299## 474.2 0.59
5-amino-N-(4- ((1R,3R,4S,5S)-3-amino- 4-methoxy-5-
methylcyclohexyl) pyridin-3-yl)-2-(2,6- difluorophenyl)thiazole-
4-carboxamide 83 ##STR00300## 524.3 0.67 N-(4-((1R,3R,4S,5S)-3-
amino-4-(2- cyanoethoxy)-5- methylcyclohexyl) pyridin-3-yl)-6-
(2,6-difluoro-4- methylphenyl)-5- fluoropicolinamide 84
##STR00301## 524.3 0.67 N-(4-((1S,3S,4R,5R)- 3-amino-4-(2-
cyanoethoxy)-5- methylcyclohexyl) pyridin-3-yl)-6- (2,6-difluoro-4-
methylphenyl)-5- fluoropicolinamide 85 ##STR00302## 510.3 0.62
N-(4-((1R,3R,4S,5S)-3- amino-4-(2- cyanoethoxy)-5-
methylcyclohexyl) pyridin-3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 86 ##STR00303## 510.3 0.62
N-(4-((1S,3S,4R,5R)-3- amino-4-(2- cyanoethoxy)-5-
methylcyclohexyl) pyridin-3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 87 ##STR00304## 483.1 0.62 N-(4-((3R,4S,5S)-
3-amino-4-azido- 5-methylpiperidin-1- yl)pyridin-3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 88 ##STR00305## 499.2 0.73
N-(4-((1R,3R,4S,5S)-3- amino-4-ethoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluorophenyl)-5- fluoropicolinamide 89
##STR00306## 499.2 0.73 N-(4-((1S,3S,4R,5R)-3- amino-4-ethoxy-5-
methylcyclohexyl) pyridin-3-yl)-6-(2,6- difluoro-4-
methylphenyl)-5- fluoropicolinamide 90 ##STR00307## 485.2 0.68
N-(4-((1S,3S,4R,5R)-3- amino-4-ethoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluorophenyl)-5- fluoropicolinamide 91
##STR00308## 485.2 0.68 N-(4-((1R,3R,4S,5S)-3- amino-4-ethoxy-5-
methylcyclohexyl) pyridin-3-yl)-6-(2,6- difluoro-4-
methylphenyl)-5- fluoropicolinamide 92 ##STR00309## 471.1 0.64
N-(4-((1R,3R,4S,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluorophenyl)-5- fluoropicolinamide 93
##STR00310## 545.3 0.64 N-(4-((1R,3R,4S,5S)-3- amino-4-methoxy-5-
methylcyclohexyl) pyridin-3-yl)-6-(2,6- difluoro-4-(2-
methoxyethoxy) phenyl)-5- fluoropicolinamide 94 ##STR00311## 603.3
0.63 N-(4-((1R,3R,4S,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(4-(1,1- dioxidotetrahydro-2H- thiopyran-4-yl)-2,6-
difluorophenyl)-5- fluoropicolinamide 95 ##STR00312## 513.2 0.79
N-(4-((1R,3R,4S,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluoro-4- isopropylphenyl)-5-
fluoropicolinamide 96 ##STR00313## 573.3 0.76
N-(4-((1R,3R,4S,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluoro-4-(4- fluorotetrahydro-2H-
pyran-4-yl)phenyl)-5- fluoropicolinamide 97 ##STR00314## 529.3 0.68
N-(4-((1R,3R,4S,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluoro-4-(2- hydroxypropan-2- yl)phenyl)-5-
fluoropicolinamide 98 ##STR00315## 541.3 0.75
N-(4-((1R,3R,4S,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluoro-4-(1- hydroxycyclobutyl) phenyl)-5-
fluoropicolinamide 99 ##STR00316## 555.3 0.66
N-(4-((1R,3R,4S,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluoro-4- (tetrahydro-2H-pyran-4-
yl)phenyl)-5- fluoropicolinamide 100 ##STR00317## 529.3 0.80
N-(4-((1R,3R,4S,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(4- (ethoxymethyl)-2,6- difluorophenyl)-5-
fluoropicolinamide 101 ##STR00318## 529.3 0.86
N-(4-((1R,3R,4S,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluoro-4- isopropoxyphenyl)-5-
fluoropicolinamide 102 ##STR00319## 485.2 0.73
N-(4-((1R,3R,4S,5S)-3- amino-4-methoxy-5-
methylcyclohexyl) pyridin-3-yl)-6-(2,6- difluoro-4-
methylphenyl)-5- fluoropicolinamide 103 ##STR00320## 485.4 0.68
N-(4-((1S,3S,4R,5R)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluoro-4- methylphenyl)-5-
fluoropicolinamide 104 ##STR00321## 502.3 0.50
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4- (methylsulfonyl)
cyclohexyl)pyridin- 3-yl)-2-(2,6- difluorophenyl) pyrimidine-4-
carboxamide 105 ##STR00322## 589.4 0.62 methyl ((3R,4S,5S)-
3-amino-1-(3-(6- (2,6-difluoro-4-(2- methoxyethoxy)phenyl)-
5-fluoropicolinamido) pyridin-4-yl)-5- methylpiperidin-4-
yl)carbamate 106 ##STR00323## 588.4 0.63 methyl ((1S,2R,4R,6S)-2-
amino-4-(3-(6-(2,6- difluoro-4-(2- methoxyethoxy)phenyl)-
5-fluoropicolinamido) pyridin-4-yl)-6- methylcyclohexyl) carbamate
107 ##STR00324## 619.4 0.65 N-(4-((1R,3R,4S,5S)-3-
amino-5-methyl-4- (methylsulfonyl) cyclohexyl)pyridin-
3-yl)-6-(2,6-difluoro-4- ((tetrahydro-2H-pyran-4- yl)oxy)phenyl)-5-
fluoropicolinamide 108 ##STR00325## 589.4 0.62
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4- (methylsulfonyl)
cyclohexyl)pyridin- 3-yl)-6-(2,6-difluoro-4- (1-hydroxycyclobutyl)
phenyl)-5- fluoropicolinamide 109 ##STR00326## 583.4 0.58
3-amino-N-(4- ((1R,3R,4S,5S)-3-amino- 5-methyl-4- (methylsulfonyl)
cyclohexyl)pyridin- 3-yl)-6-(2-fluoro-5- (isopropylcarbamoyl)
phenyl) picolinamide 110 ##STR00327## 621.4 0.66
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4- (methylsulfonyl)
cyclohexyl)pyridin- 3-yl)-6-(2,6-difluoro-4-
(4-fluorotetrahydro-2H- pyran-4-yl)phenyl)-5- fluoropicolinamide
111 ##STR00328## 574.4 0.59 3-amino-N-(4- ((1R,3R,4S,5S)-3-amino-4-
(2-cyanoethoxy)-5- methylcyclohexyl) pyridin-3-yl)-6- (2-fluoro-5-
(isopropylcarbamoyl) phenyl) picolinamide 112 ##STR00329## 549.3
0.63 N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4- (methylsulfonyl)
cyclohexyl)pyridin- 3-yl)-6-(2,6-difluoro-4- methoxyphenyl)-5-
fluoropicolinamide 113 ##STR00330## 534.2 0.69
N-(4-((1S,3S,4R,5R)-3- amino-5-methyl-4-
(pyridin-2-yloxy)cyclohexyl) pyridin-3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 114 ##STR00331## 534.3 0.61
3-amino-N-(4- ((1R,3R,4S,5S)-3-amino- 5-methyl-4- (methylsulfonyl)
cyclohexyl)pyridin- 3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 115 ##STR00332## 522.2 0.57 5-amino-N-(4-
((1R,3R,4S,5S)-3- amino-5-methyl-4- (methylsulfonyl)
cyclohexyl)pyridin- 3-yl)-2-(2,6- difluorophenyl)thiazole-4-
carboxamide 116 ##STR00333## 577.3 0.66 N-(4-((1R,3R,4S,5S)-3-
amino-5-methyl-4- (methylsulfonyl) cyclohexyl)pyridin- 3-yl)-6-(4-
(ethoxymethyl)-2,6- difluorophenyl)-5- fluoropicolinamide 117
##STR00334## 571.4 0.59 N-(4-((1R,3R,4R,5S)-3- amino-4-methoxy-5-
methylcyclohexyl) pyridin-3-yl)-6-(2,6- difluoro-4-(4-
hydroxytetrahydro-2H- pyran-4-yl)phenyl)-5- fluoropicolinamide 118
##STR00335## 663.5 0.58 N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-(2-
(methylsulfonyl)ethoxy) cyclohexyl)pyridin- 3-yl)-6-(2,6-
difluoro-4-(4- hydroxytetrahydro-2H- pyran-4-yl)phenyl)-5-
fluoropicolinamide 119 ##STR00336## 593.3 0.62
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4- (methylsulfonyl)
cyclohexyl)pyridin- 3-yl)-6-(2,6-difluoro-4- (2-methoxyethoxy)
phenyl)-5- fluoropicolinamide 120 ##STR00337## 507.2 0.55
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-
(methylsulfonyl)cyclohexyl) pyridin-3-yl)-2-(2,6-
difluorophenyl)thiazole-4- carboxamide 121 ##STR00338## 619.4 0.56
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-
(methylsulfonyl)cyclohexyl) pyridin-3-yl)-6-(2,6- difluoro-4-
(4-hydroxytetrahydro- 2H-pyran-4-yl)phenyl)-5- fluoropicolinamide
122 ##STR00339## 533.4 0.63 N-(4-((1R,3R,4S,5S)-3-
amino-5-methyl-4- (methylsulfonyl)cyclohexyl) pyridin-3-yl)-6-(2,6-
difluoro-4- methylphenyl)-5- fluoropicolinamide 123 ##STR00340##
534.4 0.69 N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-
(pyridin-2-yloxy) cyclohexyl)pyridin- 3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 124 ##STR00341## 545.3 0.67
N-(4-((1R,3R,4R,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluoro-4-(2- methoxyethoxy) phenyl)-5-
fluoropicolinamide 125 ##STR00342## 621.3 0.72
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-(2- (methylsulfonyl)ethoxy)
cyclohexyl)pyridin-3- yl)-6-(4- (ethoxymethyl)-2,6-
difluorophenyl)-5- fluoropicolinamide 126 ##STR00343## 535.4 0.61
3-amino-N-(4- ((1R,3R,4R,5S)-3- amino-4-methoxy-5-
methylcyclohexyl) pyridin-3-yl)-6-(2- fluoro-5-
(isopropylcarbamoyl) phenyl) picolinamide 127 ##STR00344## 573.3
0.70 N-(4-((1R,3R,4R,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluoro-4-(4- fluorotetrahydro-2H-
pyran-4-yl)phenyl)-5- fluoropicolinamide 128 ##STR00345## 627.4
0.59 3-amino-N-(4- ((1R,3R,4S,5S)-3- amino-5-methyl-4-(2-
(methylsulfonyl)ethoxy) cyclohexyl)pyridin-3-yl)- 6-(2-fluoro-5-
(isopropylcarbamoyl) phenyl) picolinamide 129 ##STR00346## 665.4
0.68 N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-(2-
(methylsulfonyl)ethoxy) cyclohexyl)pyridin- 3-yl)-6-(2,6-
difluoro-4-(4- fluorotetrahydro-2H- pyran-4-yl)phenyl)-5-
fluoropicolinamide 130 ##STR00347## 637.4 0.68
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-(2- (methylsulfonyl)ethoxy)
cyclohexyl)pyridin- 3-yl)-6-(2,6- difluoro-4-(2-
methoxyethoxy)phenyl)- 5-fluoropicolinamide 131 ##STR00348## 497.2
0.62 N-(4-((3R,4S,5S)- 3-amino-5-methyl- 4-(1H-1,2,3-triazol-1-
yl)piperidin-1-yl) pyridin-3-yl)-2-(2,6- difluorophenyl)thiazole-4-
carboxamide 132 ##STR00349## 583.3 0.68 N-(4-((3R,4S,5S)-3-amino-5-
methyl-4-(1H-1,2,3-triazol-1- yl)piperidin-1-yl)pyridin-3-yl)-
6-(2,6-difluoro-4-(2- methoxyethoxy)phenyl)-5- fluoropicolinamide
133 ##STR00350## 551.1 0.61 N-(4-((1R,3R,4S,5S)-3-
amino-5-methyl-4-(2- (methylsulfonyl)ethoxy) cyclohexyl)pyridin-
3-yl)-2-(2,6- difluorophenyl) thiazole-4- carboxamide 134
##STR00351## 459.3 0.69 N-(4-((1R,3R,4R,5S)-3- amino-4-methoxy-5-
methylcyclohexyl) pyridin-3-yl)-2-(2,6- difluorophenyl) thiazole-4-
carboxamide 135 ##STR00352## 566.2 0.63 5-amino-N-(4-
((1R,3R,4S,5S)-3-amino- 5-methyl-4-(2- (methylsulfonyl)ethoxy)
cyclohexyl)pyridin- 3-yl)-2-(2,6- difluorophenyl) thiazole-4-
carboxamide 136 ##STR00353## 529.3 0.74 N-(4-((1R,3R,4R,5S)-3-
amino-4-methoxy-5- methylcyclohexyl) pyridin-3-yl)-6-(4-
(ethoxymethyl)-2,6- difluorophenyl)-5- fluoropicolinamide 137
##STR00354## 571.4 0.67 N-(4-((1R,3R,4R,5S)-3- amino-4-methoxy-5-
methylcyclohexyl) pyridin-3-yl)-6-(2,6- difluoro-4-
((tetrahydro-2H-pyran-4- yl)oxy)phenyl)-5- fluoropicolinamide 138
##STR00355## 541.3 0.64 N-(4-((1R,3R,4R,5S)-3- amino-4-methoxy-5-
methylcyclohexyl) pyridin-3-yl)-6-(2,6- difluoro-4-(1-
hydroxycyclobutyl) phenyl)-5- fluoropicolinamide 139 ##STR00356##
633.4 0.63 N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-(2-
(methylsulfonyl)ethoxy) cyclohexyl)pyridin- 3-yl)-6-(2,6-
difluoro-4-(1- hydroxycyclobutyl) phenyl)-5- fluoropicolinamide 140
##STR00357## 663.4 0.66 N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-(2-
(methylsulfonyl)ethoxy) cyclohexyl)pyridin- 3-yl)-6-(2,6-
difluoro-4-((tetrahydro- 2H-pyran-4-yl) oxy)phenyl)-5-
fluoropicolinamide 141 ##STR00358## 621.4 0.81
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-(2- (methylsulfonyl)ethoxy)
cyclohexyl)pyridin- 3-yl)-6-(2,6- difluoro-4- isopropoxyphenyl)-
5-fluoropicolinamide 142 ##STR00359## 529.3 0.82
N-(4-((1R,3R,4R,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluoro-4- isopropoxyphenyl)-5-
fluoropicolinamide 143 ##STR00360## 501.1 0.70
N-(4-((1R,3R,4R,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluoro-4- methoxyphenyl)-5-
fluoropicolinamide 144 ##STR00361## 593.3 0.68
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-(2- (methylsulfonyl)ethoxy)
cyclohexyl)pyridin- 3-yl)-6-(2,6-difluoro-4- methoxyphenyl)-5-
fluoropicolinamide 145 ##STR00362## 486.1 0.69 3-amino-N-(4-
((1R,3R,4R,5S)-3- amino-4-methoxy-5- methylcyclohexyl)
pyridin-3-yl)-6-(2,6- difluorophenyl)-5- fluoropicolinamide 146
##STR00363## 578.2 0.66 3-amino-N-(4- ((1R,3R,4S,5S)-3-amino-
5-methyl-4-(2- (methylsulfonyl)ethoxy) cyclohexyl)pyridin-
3-yl)-6-(2,6- difluorophenyl)-5- fluoropicolinamide 147
##STR00364## 567.3 0.72 N-(4-((3R,4S,5S)- 3-amino-5-rnethyl-
4-(1H-1,2,3-triazol-1- yl)piperidin-1-yl) pyridin-3-yl)-
6-(2,6-difluoro-4- isopropoxyphenyl)-5- fluoropicolinamide 148
##STR00365## 573.3 0.72 methyl (3R,4S,5S)- 3-amino-1-(3-(6-(2,6-
difluoro-4- isopropoxyphenyl)- 5-fluoropicolinamido)
pyridin-4-yl)-5- methylpiperidin-4- ylcarbamate 149 ##STR00366##
572.3 0.74 methyl (1S,2R,4R,6S)- 2-amino-4-(3-(6- (2,6-difluoro-4-
isopropoxyphenyl)- 5-fluoropicolinamido) pyridin-4-yl)-6-
methylcyclohexyl- carbamate 150 ##STR00367## 534.4 0.60
N-(4-((1S,3S,4R,5R)-3- amino-5-methyl-4-(2- oxopyridin-1(2H)-
yl)cyclohexyl)pyridin- 3-yl)-6- (2,6-difluorophenyl)-5-
fluoropicolinamide 151 ##STR00368## 611.4 0.67 N-(4-((3R,4S,5S)-
3-amino-5-methyl- 4-(1H-1,2,3-triazol-1- yl)piperidin-1-yl)
pyridin-3-yl)-6- (2,6-difluoro-4-(4- fluorotetrahydro-2H-
pyran-4-yl)phenyl)-5- fluoropicolinamide 152 ##STR00369## 534.4
0.60 N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-(2- oxopyridin-1(2H)-
yl)cyclohexyl)pyridin- 3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 153 ##STR00370## 612.4 0.70 N-(4-((1R,3R,4S,5S)-
3-amino-4-(2- cyanoethoxy)-5- methylcyclohexyl) pyridin-3-yl)-6-
(2,6-difluoro-4-(4- fluorotetrahydro-2H- pyran-4-yl)phenyl)-5-
fluoropicolinamide 154 ##STR00371## 511.2 0.58 5-amino-N-(4-
((1R,3R,4S,5S)-3- amino-5-methyl-4- (1H-1,2,3-triazol-1-
yl)cyclohexyl)pyridin- 3-yl)-2-(2,6- difluorophenyl)thiazole-
4-carboxamide 155 ##STR00372## 566.3 0.68 N-(4-((1S,3R,4S,5S)-
3-amino-5-methyl- 4-(1H-1,2,3-triazol-1- yl)cyclohexyl)pyridin-
3-yl)-6-(4- (ethoxymethyl)-2,6- difluorophenyl)-5-
fluoropicolinamide 156 ##STR00373## 512.2 0.56 5-amino-N-(4-
((3R,4S,5S)-3-amino- 5-methyl-4-(1H-1,2,3-
triazol-1-yl)piperidin-1- yl)pyridin-3-yl)-2-(2,6-
difluorophenyl)thiazole-4- carboxamide 157 ##STR00374## 567.3 0.67
N-(4-((3R,4S,5S)- 3-amino-5-methyl- 4-(1H-1,2,3-triazol-1-
yl)piperidin-1-yl) pyridin-3-yl)-6-(4- (ethoxymethyl)-2,6-
difluorophenyl)-5- fluoropicolinamide 158 ##STR00375## 585.4 0.62
methyl (3R,4S,5S)- 3-amino-1-(3-(6-(2,6- difluoro-4-(1-
hydroxycyclobutyl) phenyl)-5- fluoropicolinamido) pyridin-4-yl)-5-
methylpiperidin-4- ylcarbamate 159 ##STR00376## 615.4 0.64 methyl
(3R,4S,5S)- 3-amino-1-(3-(6- (2,6-difluoro-4- (tetrahydro-2H-pyran-
4-yloxy)phenyl)-5- fluoropicolinamido) pyridin-4-yl)-5-
methylpiperidin-4- ylcarbamate 160 ##STR00377## 609.4 0.59
N-(4-((3R,4S,5S)- 3-amino-5-methyl- 4-(1H-1,2,3-triazol-1-
yl)piperidin-1-yl) pyridin-3-yl)- 6-(2,6-difluoro-4-(4-
hydroxytetrahydro- 2H-pyran-4-yl) phenyl)-5- fluoropicolinamide 161
##STR00378## 523.3 0.65 N-(4-((3R,4S,5S)- 3-amino-5-
methyl-4-(1H-1,2,3- triazol-1-yl)piperidin- 1-yl)pyridin-3-yl)-
6-(2,6-difluoro-4- methylphenyl)-5- fluoropicolinamide 162
##STR00379## 571.3 0.58 methyl (3R,4S,5S)- 3-amino-1-(3-
(6-(2,6-difluoro-4- (oxetan-3-yl)phenyl)- 5-fluoropicolinamido)
pyridin-4-yl)-5- methylpiperidin-4- ylcarbamate 163 ##STR00380##
566.3 0.61 N-(4-((1R,3R,4S,5S)-3- amino-4-(2-cyanoethoxy)-
5-methylcyclohexyl) pyridin-3-yl)-6- (2,6-difluoro-4-(oxetan-
3-yl)phenyl)-5- fluoropicolinamide 164 ##STR00381## 565.3 0.48
N-(4-((3R,4S,5S)- 3-amino-5-methyl-4-(1H-
1,2,3-triazol-1- yl)piperidin-1-yl) pyridin-3-yl)-6-
(2,6-difluoro-4-(oxetan- 3-yl)phenyl)-5- fluoropicolinamide 165
##STR00382## 540.1 0.67 N-(4-((1R,3R,4S,5S)-3-
amino-4-(2-cyanoethoxy)- 5-methylcyclohexyl) pyridin-3-yl)-6-
(2,6-difluoro-4- methoxyphenyl)-5- fluoropicolinamide 166
##STR00383## 525.1 0.66 3-amino-N-(4- ((1R,3R,4S,5S)-3- amino-4-(2-
cyanoethoxy)-5- methylcyclohexyl) pyridin-3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 167 ##STR00384## 584.4 0.63
methyl (1S,2R,4R,6S)-2- amino-4-(3-(6-(2,6- difluoro-4-(1-
hydroxycyclobutyl) phenyl)-5- fluoropicolinamido) pyridin-4-yl)-6-
methylcyclohexyl- carbamate 168 ##STR00385## 582.3 0.64
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4- (1H-1,2,3-triazol-1-
yl)cyclohexyl) pyridin-3-yl)-6- (2,6-difluoro-4-(2- methoxyethoxy)
phenyl)-5- fluoropicolinamide 169 ##STR00386## 496.2 0.57
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl- 4-(1H-1,2,3-triazol-1-
yl)cyclohexyl) pyridin-3-yl)-2- (2,6-difluorophenyl) thiazole-
4-carboxamide 170 ##STR00387## 614.4 0.66 methyl (1S,2R,4R,6S)-2-
amino-4-(3-(6-(2,6- difluoro-4- (tetrahydro-2H-pyran-
4-yloxy)phenyl)-5- fluoropicolinamido) pyridin-4-yl)-6-
methylcyclohexyl- carbamate 171 ##STR00388## 608.4 0.65
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4- (1H-1,2,3-triazol-1-
yl)cyclohexyl) pyridin-3-yl)-6- (2,6-difluoro-4-
(tetrahydro-2H-pyran- 4-yloxy)phenyl)-5- fluoropicolinamide 172
##STR00389## 578.4 0.62 N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4-
(1H-1,2,3-triazol-1- yl)cyclohexyl) pyridin-3-yl)-6-
(2,6-difluoro-4-(1- hydroxycyclobutyl) phenyl)-5-
fluoropicolinamide 173 ##STR00390## 545.3 0.64 methyl
((3R,4S,5S)-3- amino-1-(3-(6-(2,6- difluoro-4- methoxyphenyl)-5-
fluoropicolinamido) pyridin-4-yl)-5- methylpiperidin-4-
yl)carbamate 174 ##STR00391## 530.3 0.62 methyl ((3R,4S,5S)-
3-amino-1-(3-(3- amino-6-(2,6- difluorophenyl)-5-
fluoropicolinamido) pyridin-4-yl)-5- methylpiperidin-4-
yl)carbamate 175 ##STR00392## 529.3 0.65 methyl (1S,2R,4R,6S)-2-
amino-4-(3-(3-amino- 6-(2,6-difluorophenyl)- 5-fluoropicolinamido)
pyridin-4-yl)-6- methylcyclohexyl- carbamate 176 ##STR00393## 523.3
0.63 3-amino-N-(4- ((1R,3R,4S,5S)-3- amino-5-methyl-4-(1H-
1,2,3-triazol-1-yl) cyclohexyl)pyridin- 3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 177 ##STR00394## 615.3 0.59
methyl (3R,4S,5S)- 3-amino-1-(3-(6- (2,6-difluoro-4-(4-
hydroxytetrahydro-2H- pyran-4-yl)phenyl)-5- fluoropicolinamido)
pyridin-4-yl)-5- methylpiperidin-4- ylcarbamate 178 ##STR00395##
614.3 0.59 methyl (1S,2R,4R,6S)-2- amino-4-(3-(6-(2,6-
difluoro-4-(4- hydroxytetrahydro-2H- pyran-4-yl)phenyl)-5-
fluoropicolinamido) pyridin-4-yl)-6- methylcyclohexyl- carbamate
179 ##STR00396## 608.3 0.58 N-(4-((1R,3R,4S,5S)-3-
amino-5-methyl-4- (1H-1,2,3-triazol-1- yl)cyclohexyl)pyridin-
3-yl)-6-(2,6-difluoro-4- (4-hydroxytetrahydro- 2H-pyran-4-yl)
phenyl)-5- fluoropicolinamide 180 ##STR00397## 528.2 0.66 methyl
(1S,2R,4R,6S)-2- amino-4-(3-(6-(2,6- difluoro-4- methylphenyl)-5-
fluoropicolinamido) pyridin-4-yl)-6- methylcyclohexyl- carbamate
181 ##STR00398## 522.2 0.65 N-(4-((1R,3R,4S,5S)-3-
amino-5-methyl-4- (1H-1,2,3-triazol-1- yl)cyclohexyl)pyridin-
3-yl)-6-(2,6-difluoro-4- methylphenyl)-5- fluoropicolinamide 182
##STR00399## 570.3 0.61 methyl (1S,2R,4R,6S)-2- amino-4-(3-(6-(2,6-
difluoro-4-(oxetan- 3-yl)phenyl)-5- fluoropicolinamido)
pyridin-4-yl)-6- methylcyclohexyl- carbamate 183 ##STR00400## 610.4
0.70 N-(4-((1R,3R,4S,5S)-3- amino-4-(2-cyanoethoxy)-
5-methylcyclohexyl) pyridin-3-yl)-6- (2,6-difluoro-4-
((tetrahydro-2H-pyran- 4-yl)oxy)phenyl)-5- fluoropicolinamide 184
##STR00401## 580.4 0.66 N-(4-((1R,3R,4S,5S)-3-
amino-4-(2-cyanoethoxy)- 5-methylcyclohexyl) pyridin-3-yl)-6-
(2,6-difluoro-4-(1- hydroxycyclobutyl) phenyl)-5-
fluoropicolinamide 185 ##STR00402## 533.3 0.66
N-(4-((1R,3R,4S,5S)-3- amino-4-(4-cyano- 1H-1,2,3-triazol-1-yl)-5-
methylcyclohexyl) pyridin-3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 186 ##STR00403## 548.4 0.67
N-(4-((1R,3R,4S,5S)-3- amino-4-(4-cyclopropyl-
1H-1,2,3-triazol-1-yl)-5- methylcyclohexyl) pyridin-3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 187 ##STR00404## 548.3 0.69
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4- (4-(prop-1-en-2-yl)-
1H-1,2,3-triazol-1- yl)cyclohexyl)pyridin- 3-yl)-6-(2,6-
difluorophenyl)-5- fluoropicolinamide 188 ##STR00405## 566.4 0.62
N-(4-((1R,3R,4S,5S)-3- amino-4-(4-(2- hydroxypropan-2-yl)-1H-
1,2,3-triazol-1-yl)-5- methylcyclohexyl) pyridin-3-yl)-6-
(2,6-difluorophenyl)-5- fluoropicolinamide 189 ##STR00406## 515.3
0.62 2-((1R,2S,4S,6R)- 2-amino-4-(3-(6-(2,6- difluorophenyl)-5-
fluoropicolinamido) pyridin-4-yl)-6- methylcyclohexyloxy) acetic
acid 190 ##STR00407## 515.3 0.62 2-((1S,2R,4R,6S)- 2-amino-4-(3-(6-
(2,6-difluorophenyl)-5- fluoropicolinamido) pyridin-4-yl)-6-
methylcyclohexyloxy) acetic acid 191 ##STR00408## 544.3 0.66 methyl
(1S,2R,4R,6S)-2- amino-4-(3-(6-(2,6- difluoro-4- methoxyphenyl)-5-
fluoropicolinamido) pyridin-4-yl)-6- methylcyclohexyl- carbamate
192 ##STR00409## 538.3 0.65 N-(4-((1R,3R,4S,5S)-3-
amino-5-methyl-4- (1H-1,2,3-triazol-1- yl)cyclohexyl)
pyridin-3-yl)-6- (2,6-difluoro-4- methoxyphenyl)-5-
fluoropicolinamide 193 ##STR00410## 539.3 0.64 N-(4-((3R,4S,5S)-3-
amino-5-methyl-4- (1H-1,2,3-triazol-1- yl)piperidin-1-yl)
pyridin-3-yl)- 6-(2,6-difluoro-4- methoxyphenyl)-5-
fluoropicolinamide 194 ##STR00411## 524.3 0.62 3-amino-N-(4-
((3R,4S,5S)-3- amino-5-methyl-4- (1H-1,2,3-triazol-1-
yl)piperidin-1- yl)pyridin-3-yl)-6-(2,6- difluorophenyl)-5-
fluoropicolinamide 195 ##STR00412## 610.4 0.67
N-(4-((1R,3R,4S,5S)-3- amino-5-methyl-4- (1H-1,2,3-triazol-1-
yl)cyclohexyl) pyridin-3-yl)-6- (2,6-difluoro-4-(4-
fluorotetrahydro-2H- pyran-4-yl)phenyl)-5- fluoropicolinamide 196
##STR00413## 572.4 0.59 3-amino-N-(4- ((1R,3R,4S,5S)-3-
amino-5-methyl-4- (1H-1,2,3-triazol-1- yl)cyclohexyl)pyridin-
3-yl)-6-(2-fluoro-5- (isopropylcarbamoyl) phenyl)picolinamide 197
##STR00414## 573.4 0.58 3-amino-N-(4- ((3R,4S,5S)-3-
amino-5-methyl-4- (1H-1,2,3-triazol-1- yl)piperidin-1-
yl)pyridin-3-yl)- 6-(2-fluoro-5- (isopropylcarbamoyl) phenyl)
picolinamide 198 ##STR00415## 579.3 0.60 methyl (3R,4S,5S)-3-
amino-1-(3-(3- amino-6-(2-fluoro-5- (isopropylcarbamoyl)
phenyl)picolinamido) pyridin-4-yl)-5- methylpiperidin-4-
ylcarbamate 199 ##STR00416## 617.4 0.68 methyl (3R,4S,5S)-3-
amino-1-(3-(6-(2,6- difluoro-4-(4- fluorotetrahydro-2H-
pyran-4-yl)phenyl)-5- fluoropicolinamido) pyridin-4-yl)-5-
methylpiperidin-4- ylcarbamate 200 ##STR00417## 578.4 0.60 methyl
(1S,2R,4R,6S)-2- amino-4-(3-(3-amino- 6-(2-fluoro-5-
(isopropylcarbamoyl) phenyl)picolinamido) pyridin-4-yl)-6-
methylcyclohexyl- carbamate 201 ##STR00418## 616.4 0.68 methyl
(1S,2R,4R,6S)-2- amino-4-(3-(6-(2,6- difluoro-4-(4-
fluorotetrahydro-2H- pyran-4-yl)phenyl)-5- fluoropicolinamido)
pyridin-4-yl)-6- methylcyclohexyl- carbamate 202 ##STR00419## 572.2
0.67 methyl (1S,2R,4R,6S)-2- amino-4-(3-(6-(4- (ethoxymethyl)-2,6-
difluorophenyl)-5- fluoropicolinamido) pyridin-4-yl)-6-
methylcyclohexyl- carbamate 203 ##STR00420## 573.2 0.66 methyl
(3R,4S,5S)-3- amino-1-(3-(6-(4- (ethoxymethyl)-
2,6-difluorophenyl)-5- fluoropicolinamido) pyridin-4-yl)-5-
methylpiperidin-4- ylcarbamate
In addition to LC/MS and LC characterization, representative
compounds were analyzed by .sup.1H-NMR. The following data in Table
3 are typical spectra of the compounds of the invention.
TABLE-US-00004 TABLE 3 Ex # .sup.1H-NMR data 1 .sup.1H NMR (400
MHz, Methanol-d) .delta. ppm 0.65 (d, J = 6.70 Hz, 1 H) 1.81 (d, J
= 13.11 Hz, 1 H) 2.00-2.20 (m, 1 H) 2.22-2.37 (m, 1 H) 2.61-2.79
(m, 1 H) 3.34-3.45 (m, 1 H) 3.89 (dt, J = 12.76, 4.30 Hz, 1 H) 4.86
(s, 5 H) 5.00 (br. s., 1 H) 7.22 (t, J = 8.31 Hz, 1 H) 7.64 (tt, J
= 8.48, 6.46 Hz, 1 H) 7.83-7.92 (m, 1 H) 8.04 (t, J = 8.73 Hz, 1 H)
8.08 (s, 1 H) 8.42 (dd, J = 8.63, 3.94 Hz, 1 H) 8.60 (br. s., 1 H)
8.90 (br. s., 1 H) 4 .sup.1H NMR (400 MHz, METHANOL-d4) .delta. ppm
0.48 (d, J = 6.90 Hz, 3 H) 2.40-2.55 (m, 1 H) 2.64 (s, 1 H)
3.49-3.61 (m, 1 H) 3.67-3.79 (m, 1 H) 3.90 (dd, J = 11.40, 4.30 Hz,
1 H) 3.97-4.08 (m, 1 H) 4.14-4.28 (m, 1 H) 4.96 (t, J = 4.38 Hz, 1
H) 7.26 (t, J = 8.39 Hz, 2 H) 7.49-7.57 (m, 3 H) 7.61-7.72 (m, 1 H)
7.82 (dd, J = 2.64, 1.52 Hz, 1 H) 8.05 (t, J = 8.71 Hz, 1 H) 8.33
(s, 1 H) 8.39 (dd, J = 6.58, 0.90 Hz, 1 H) 8.46 (dd, J = 8.68, 3.94
Hz, 1 H) 9.15 (s, 1 H) 5 .sup.1H NMR (400 MHz, METHANOL-d4) .delta.
ppm 0.41 (d, J = 6.85 Hz, 3 H) 2.50 (td, J = 11.57, 4.55 Hz, 1 H)
3.59 (t, J = 12.94 Hz, 1 H) 3.81 (dd, J = 13.60, 3.81 Hz, 1 H)
3.94-4.15 (m, 2 H) 4.18-4.33 (m, 1 H) 5.11 (t, J = 4.16 Hz, 1 H)
7.29 (t, J = 8.49 Hz, 2 H) 7.62 (d, J = 6.75 Hz, 1 H) 7.70 (tt, J =
8.49, 6.50 Hz, 1 H) 7.90 (d, J = 0.93 Hz, 1 H) 8.07 (t, J = 8.68
Hz, 1 H) 8.17 (d, J = 0.68 Hz, 1 H) 8.42 (d, J = 6.65 Hz, 1 H) 8.48
(dd, J = 8.66, 3.96 Hz, 1 H) 9.13 (s, 1 H) 6 .sup.1H NMR in DMSOd6:
.delta. 10.94 (s, 1H), 10.49 (s, 1H), 8.60 (s, 1H), 8.48 (d, J =
4.0, 1H), 8.35 (dd, J = 8.0, 4.0, 1H), 8.20 (t, J = 8.0, 1 H), 8.02
(broad doublet, J = 4.0, 2H), 7.67-7.74 (m, 1H), 7.42 (d, J = 4.0,
1H), 7.36 (t, J = 8.0, 2H), 4.24 (m, 1H), 3.82-3.86 (m, 1H),
3.21-3.27 (m, 1H), 2.50-2.55 (m, 1H), 2.24 (d, J = 12.0, 1H), 1.86
(d, J = 16.0, 1H), 1.68 (q, J = 12.0, 1H), 1.59 (q, J = 12.0, 1H),
1.40 (d, J = 8.0, 3H). 7 .sup.1H NMR (400 MHz, METHANOL-d4) .delta.
ppm 0.42 (d, J = 6.85 Hz, 3 H) 2.37 (s, 3 H) 2.46 (dd, J = 11.71,
7.02 Hz, 1 H) 3.55 (t, J = 12.84 Hz, 1 H) 3.77 (dd, J = 13.62, 3.99
Hz, 1 H) 3.89-4.09 (m, 2 H) 4.12-4.30 (m, 1 H) 4.93 (t, J = 4.40
Hz, 1 H) 7.27 (t, J = 8.41 Hz, 2 H) 7.56 (d, J = 6.75 Hz, 1 H) 7.68
(tt, J = 8.49, 6.48 Hz, 1 H) 7.83 (d, J = 0.59 Hz, 1 H) 8.06 (t, J
= 8.71 Hz, 1 H) 8.36-8.42 (m, 1 H) 8.47 (dd, J = 8.68, 3.94 Hz, 1
H) 9.14 (s, 1 H) 11 .sup.1H NMR in DMSO.sub.d6: .delta. 10.55 (s,
1H), 8.55 (s, 1H), 8.47 (d, J = 4.0, 1H), 8.37 (dd, J = 8.0, 4.0,
1H), 8.21 (t, J = 8.0, 1H), 8.16 (broad doublet, J = 4.0, 2H),
7.67-7.74 (m, 1H), 7.40 (d, J = 8.0, 1H), 7.36 (t, J = 8.0, 2H),
4.20-4.26 (m, 1H), 3.50-3.70 (m, 2H), 2.76-2.82 (m, 1H), 2.49-2.54
(m, 1H), 2.32-2.36 (m, 1H), 2.16-2.18 (m, 1 H), 1.91 (q, J = 12,
1H), 1.65 (q, J = 12, 1H), 0.97 (d, J = 8.0, 3H). 12 .sup.1H NMR in
DMSO.sub.d6: .delta. 10.49 (s, 1H), 8.57 (s, 1H), 8.47 (d, J = 4.0,
1H), 8.35 (dd, J = 8.0, 4.0, 1H), 8.25 (broad doublet, J = 4.0,
2H), 8.20 (t, J = 8.0, 1H), 7.67-7.74 (m, 1H), 7.42 (d, J = 8.0,
1H), 7.36 (t, J = 8.0, 2H), 4.04-4.08 (m, 1H), 3.79 (s, 3H),
3.23-3.29 (m, 1H), 2.39-2.45 (m, 1H), 2.11 (d, J = 8.0, 1H), 2.10
(d, J = 8.0, 1H), 1.90 (q, J = 12, 1H), 1.40 (q, J = 12, 1H), 1.01
(d, J = 4.0, 3H). 13 .sup.1H NMR in DMSO.sub.d6: .delta. 10.48 (s,
1H), 8.57 (s, 1H), 8.47 (d, J = 4.0, 1H), 8.35 (dd, J = 8.0, 4.0,
1H), 8.20 (t, J = 8.0, 1H), 8.08 (broad singlet, 2H), 7.67-7.74 (m,
1H), 7.42 (d, J = 8.0, 1H), 7.36 (t, J = 8.0, 2H), 3.88-3.92 (m,
1H), 3.80 (s, 3H), 3.22-3.28 (m, 1H), 2.51-2.58 (m, 1H), 2.25 (d, J
= 12.0, 1H), 1.86 (d, J = 12.0, 1H), 1.70 (q, J = 12, 1H), 1.62 (q,
J = 12, 1H), 1.34 (d, J = 4.0, 3H). 14 .sup.1H NMR (400 MHz,
METHANOL-d4) .delta. ppm 0.43 (d, J = 6.85 Hz, 3 H) 2.36-2.53 (m, 1
H) 3.43 (s, 3 H) 3.46-3.58 (m, 1 H) 3.69 (dd, J = 13.57, 3.79 Hz, 1
H) 3.88 (dd, J = 12.30, 4.52 Hz, 1 H) 4.02 (dt, J = 11.63, 4.68 Hz,
1 H) 4.12-4.26 (m, 1 H) 4.60 (s, 2 H) 4.95 (t, J = 4.38 Hz, 2 H)
7.28 (t, J = 8.39 Hz, 2 H) 7.54 (d, J = 6.55 Hz, 1 H) 7.69 (tt, J =
8.50, 6.47 Hz, 1 H) 8.00-8.12 (m, 2 H) 8.39 (dd, J = 6.55, 0.88 Hz,
1 H) 8.47 (dd, J = 8.66, 3.96 Hz, 1 H) 9.18 (s, 1 H) 15 .sup.1H NMR
in DMSO.sub.d6: .delta. 10.51 (s, 1H), 8.53 (s, 1H), 8.51 (d, J =
4.0, 1H), 8.33 (dd, J = 8.0, 4.0, 1H), 8.20 (t, J = 8.0, 1H), 8.09
(broad doublet, J = 4.0, 2H), 7.65-7.75 (m, 1H), 7.36 (t, J = 8.0,
2H), 7.30 (d, J = 8.0, 1H), 3.84-4.02 (m, 3H), 3.58-3.68 (m, 1H),
3.43-3.53 (m, 1H), 3.28-3.36 (m, 1H), 3.04-3.14 (m, 1H), 1.92-2.18
(m, 4 H), 1.56-1.63 (m, 1H), 1.24 (d, J = 8.0, 3H). 30 (.sup.1H
NMR, DMSO.sub.d-6) .delta. 10.48 (s, 1H), 8.57 (s, 1H), 8.48 (d, J
= 8.0, 1H), 8.33 (dd, J = 8.0, 4.0, 1H), 8.20 (t, J = 8.0, 1H),
7.80 (bs, 2H), 7.71 (quintet, J = 8.0, 2H), 7.43 (d, J = 4.0, 1H),
7.37 (t, J = 8.0, 2H), 3.43-3.50 (m, 2H), 2.88-2.96 (m, 1H), 2.04
(d, J = 8.0, 1H), 1.90 (s, 3H), 1.82 (d, J = 12.0, 1H), 1.64 (q, J
= 12.0, 1H), 1.46-1.55 (m, 1H), 1.32 (q, J = 12.0, 1H), 0.85 (d, J
= 4.0, 3H). 33 .sup.1H NMR, 400 MHz DMSOd6, .delta. 10.44 (s, 1H),
8.58 (d, J = 4.0, 1H), 8.47 (d, J = 4.0, 1H), 8.34 (dd, J = 8.0,
4.0, 1H), 8.20 (dd, J = 8.0, 8.0, 1H), 8.20 (dd, J = 16.0, 4.0,
1H), 7.67-7.74 (m, 1H), 7.36 (dd, J = 8.0, 8.0, 2H), 7.26 (d, J =
4.0, 1H), 2.85-2.95 (m, 2H), 2.18 (s, 3H), 1.88-1.98 (m, 1H),
1.74-1.84 (m, 1H), 1.48-1.56 (m, 1H), 1.38-1.48 (m, 1H), 1.18-1.28
(m, 1H), 1.02 (d, J = 8.0, 3H) 34 .sup.1H NMR, 400 MHz DMSOd6,
.delta. 10.44 (s, 1H), 8.60 (d, J = 8.0, 1H), 8.48 (d, J = 4.0,
1H), 8.34 (dd, J = 8.0, 4.0, 1H), 8.20 (dd, J = 8.0, 8.0, 1H), 8.00
(dd, J = 16.0, 4.0, 1H), 7.67-7.74 (m, 1H), 7.34-7.38 (m, 3H), 3.14
(s, 3H), 3.02-3.12 (m, 2H), 2.18-2.24 (m, 1H), 1.84-1.96 (m, 3H),
1.62-1.72 (m, 1H), 1.38-1.48 (m, 1H), 1.18 (d, J = 4.0, 3H). 36
.sup.1H NMR, 400 MHz DMSOd6, .delta. 10.48 (s, 1H), 8.58 (d, J =
4.0, 1H), 8.50 (d, J = 4.0, 1H), 8.34 (dd, J = 8.0, 4.0, 1H), 8.20
(dd, J = 8.0, 8.0, 1H), 8.20 (dd, J = 16.0, 4.0, 1H), 7.67-7.74 (m,
1H), 7.42 (d, J = 4.0, 1H), 7.36 (dd, J = 8.0, 8.0, 2H), 3.40-3.42
(m, 1H), 3.06-3.20 (m, 1H), 2.92 (s, 3H), 2.06-2.20 (m, 1H),
1.95-2.04 (m, 2H), 1.70-1.80 (m, 1H), 1.56-1.70 (m, 1H), 0.86 (d, J
= 8.0, 3H). 37 .sup.1H NMR, 400 MHz DMSOd6, .delta. 10.52 (s, 1H),
8.51-8.54 (m, 2H), 8.34 (dd, J = 8.0, 4.0, 1H), 8.20 (dd, J = 8.0,
8.0, 1H), 8.15 (dd, J = 16.0, 4.0, 1H), 7.67-7.74 (m, 1H), 7.36
(dd, J = 8.0, 8.0, 2H), 7.28 (d, J = 4.0, 1H), 3.77-3.79 (m, 1H),
3.18 (s, 3H), 3.02-3.20 (m, 1H), 1.94-2.40 (m, 4H), 1.57-1.62 (m,
1H), 1.24 (d, J = 8.0, 3H). 56 .sup.1H NMR, (400, DMSOd6) .delta.
10.45 (s, 1H), 8.92 (s, 1H), 8.56 (s, 1H), 8.43 (d, J = 4.0, 1H),
8.40 (dd, J = 8.0, 4.0, 1H), 8.24 (t, J = 8.0, 1H), 8.17-8.24 (m,
2H), 8.17 (s, 1H), 7.70-7.80 (m, 1H), 7.35-7.43 (m, 3H), 4.780-4.82
(m, 1H), 3.96 (t, J = 12.0, 1H), 3.60-3.90 (m, 3H), 3.50 (t, J =
12.0, 1H), 2.25-2.35 (m, 1H), 0.31 (d, J = 8.0, 3H). 64 .sup.1H
NMR, (400 MHz, METHANOL-d4) .delta. ppm 1.19 (d, J = 6.90 Hz, 3 H)
1.42 (d, J = 12.76 Hz, 1 H) 1.63-1.77 (m, 2 H) 1.84-2.04 (m, 3 H)
2.71-2.91 (m, 2 H) 3.13-3.24 (m, 2 H) 3.40-3.52 (m, 1 H) 3.73 (s, 1
H) 3.76-3.87 (m, 1 H) 3.93 (td, J = 8.68, 5.14 Hz, 1 H) 7.14 (t, J
= 8.75 Hz, 2 H) 7.47 (tt, J = 8.44, 6.25 Hz, 1 H) 7.68 (d, J = 5.62
Hz, 1 H) 8.46 (d, J = 5.62 Hz, 1 H) 9.18 (s, 1 H) 66 .sup.1H NMR,
(400 MHz, METHANOL-d4) .delta. ppm 0.43 (d, J = 6.80 Hz, 3 H)
2.37-2.60 (m, 1 H) 3.60 (t, J = 12.94 Hz, 1 H) 3.83 (dd, J = 13.64,
4.11 Hz, 1 H) 3.95-4.14 (m, 2 H) 4.17-4.32 (m, 1 H) 4.75 (s, 2 H)
5.03 (t, J = 4.01 Hz, 1 H) 7.27 (t, J = 8.46 Hz, 2 H) 7.59 (d, J =
6.85 Hz, 1 H) 7.62-7.75 (m, 1 H) 7.97-8.12 (m, 2 H) 8.40 (dd, J =
6.80, 0.93 Hz, 1 H) 8.47 (dd, J = 8.66, 3.91 Hz, 1 H) 9.12 (d, J =
0.83 Hz, 1 H). 74 .sup.1H NMR, (400 MHz, DMSO-d6) .delta. ppm 0.96
(d, J = 6.85 Hz, 3 H) 1.16-1.36 (m, 1 H) 1.46 (d, J = 12.81 Hz, 1
H) 1.50-1.78 (m, 3 H) 2.41 (s, 3 H) 2.89 (t, J = 12.06 Hz, 1 H)
3.07 (s, 3 H) 3.24 (br. s., 2 H) 3.86 (dt, J = 10.20, 5.12 Hz, 2 H)
3.99 (td, J = 9.29, 4.30 Hz, 1 H) 7.17 (d, J = 9.29 Hz, 2 H) 7.22
(d, J = 5.23 Hz, 1 H) 7.86 (br. s., 3 H) 8.08-8.20 (m, 1 H) 8.28
(dd, J = 8.66, 4.06 Hz, 1 H) 8.43 (d, J = 5.18 Hz, 1 H) 8.58 (s, 1
H) 10.38 (s, 1 H). 77 .sup.1H NMR, (400 MHz, DMSO-d6) .delta. ppm
1.00 (d, J = 6.80 Hz, 3 H) 1.42-1.81 (m, 3 H) 2.92 (t, J = 12.32
Hz, 1 H) 3.10 (s, 2 H) 3.28 (br. s., 1 H) 3.33-3.44 (m, 1 H) 3.54
(br. s., 1 H) 3.59-3.72 (m, 1 H) 3.89 (dt, J = 10.17, 5.09 Hz, 1 H)
4.02 (td, J = 9.26, 4.23 Hz, 1 H) 7.23 (d, J = 5.23 Hz, 1 H) 7.36
(t, J = 8.31 Hz, 2 H) 7.63-7.77 (m, 1 H) 7.89 (br. s., 2 H)
8.13-8.27 (m, 1 H) 8.33 (dd, J = 8.68, 4.08 Hz, 1 H) 8.46 (d, J =
5.13 Hz, 1 H) 8.60 (s, 1 H) 10.43 (s, 1 H). 86 .sup.1H NMR, (400
MHz, METHANOL-d4) .delta. ppm 1.13 (d, J = 6.80 Hz, 3 H) 1.46-1.86
(m, 3 H) 1.88-2.06 (m, 2 H) 2.72-2.93 (m, 2 H) 3.08-3.21 (m, 1 H)
3.34-3.42 (m, 1 H) 3.70 (s, 1 H) 3.76-3.88 (m, 1 H) 3.93 (td, J =
8.69, 5.01 Hz, 1 H) 7.22 (t, J = 8.27 Hz, 2 H) 7.55-7.77 (m, 2 H)
8.04 (t, J = 8.71 Hz, 1 H) 8.42 (dd, J = 8.66, 3.96 Hz, 1 H) 8.55
(d, J = 5.62 Hz, 1 H) 9.03 (s, 1 H) 90 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 8.95 (s, 1 H), 8.50 (d, J = 5.48 Hz, 1
H), 8.39 (dd, J = 8.61, 3.91 Hz, 1 H), 8.00 (t, J = 8.80 Hz, 1 H),
7.60 (d, J = 5.48 Hz, 1 H), 7.05 (d, J = 9.39 Hz, 2 H), 3.72 (quin,
J = 7.24 Hz, 2 H), 3.57 (br. s., 1 H), 3.08 (t, J = 11.74 Hz, 1 H),
2.47 (s, 3 H), 1.84-2.00 (m, 2 H), 1.72 (d, J = 5.48 Hz, 1 H),
1.54-1.65 (m, 2 H), 1.28 (t, J = 7.04 Hz, 3 H), 1.06 (d, J = 6.65
Hz, 3 H) 91 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 8.96
(s, 1 H), 8.50 (d, J = 5.48 Hz, 1 H), 8.41 (dd, J = 8.61, 3.91 Hz,
1 H), 8.03 (t, J = 8.80 Hz, 1 H), 7.58-7.68 (m, 2 H), 7.21 (t, J =
8.41 Hz, 2 H), 3.72 (quin, J = 7.24 Hz, 2 H), 3.57 (br. s., 1 H),
3.03-3.15 (m, 1 H), 1.84-2.00 (m, 2 H), 1.72 (br. s., 1 H),
1.51-1.66 (m, 2 H), 1.28 (t, J = 7.04 Hz, 3 H), 1.06 (d, J = 6.65
Hz, 3 H) 92 .sup.1H NMR, (300 MHz, <cd3od>) .delta. ppm
8.93-9.05 (m, 1 H), 8.46-8.57 (m, 1 H), 8.35-8.46 (m, 1 H),
7.97-8.09 (m, 1 H), 7.55-7.73 (m, 2 H), 7.15-7.30 (m, 2 H), 3.58
(s, 3 H), 3.42-3.51 (m, 1 H), 3.00-3.18 (m, 1 H), 1.83-2.00 (m, 2
H), 1.45-1.81 (m, 3 H), 1.07 (d, J = 6.74 Hz, 3 H) 94 .sup.1H NMR,
(400 MHz, METHANOL-d4) .delta. ppm 8.82 (s, 1 H), 8.48 (d, J = 5.48
Hz, 1 H), 8.39 (dd, J = 8.61, 3.91 Hz, 1 H), 8.01 (t, J = 8.61 Hz,
1 H), 7.54 (d, J = 5.09 Hz, 1 H), 7.19 (d, J = 9.39 Hz, 2 H), 3.58
(s, 3 H), 3.46 (s, 1 H), 3.00-3.21 (m, 5 H), 2.23-2.40 (m, 4 H),
1.85-1.95 (m, 2 H), 1.75 (br. s., 1 H), 1.50-1.67 (m, 2 H), 1.08
(d, J = 6.65 Hz, 3 H) 95 .sup.1H NMR, (400 MHz, METHANOL-d4)
.delta. ppm 8.92 (s, 1 H), 8.47 (d, J = 5.48 Hz, 1 H), 8.36 (dd, J
= 8.63, 3.94 Hz, 1 H), 7.98 (t, J = 8.71 Hz, 1 H), 7.56 (d, J =
5.48 Hz,
1 H), 7.08 (d, J = 9.44 Hz, 2 H), 3.56 (s, 3 H), 3.44 (s, 1 H),
2.95-3.10 (m, 2 H), 1.81-1.97 (m, 2 H), 1.71 (d, J = 5.97 Hz, 1 H),
1.46-1.66 (m, 2 H) 1.30 (d, J = 6.94 Hz, 6 H), 1.05 (d, J = 6.80
Hz, 3 H) 96 .sup.1H NMR, (400 MHz, METHANOL-d4) .delta. ppm 8.92
(s, 1 H), 8.49 (d, J = 5.53 Hz, 1 H), 8.39 (dd, J = 8.68, 3.94 Hz,
1 H), 8.01 (t, J = 8.73 Hz, 1 H), 7.60 (d, J = 5.58 Hz, 1 H), 7.30
(d, J = 9.24 Hz, 2 H), 3.90-4.01 (m, 2 H), 3.84 (td, J = 11.91,
1.61 Hz, 2 H), 3.44 (s, 1 H), 2.99-3.15 (m, 1 H), 2.30 (td, J =
13.14, 5.45 Hz, 1 H), 2.20 (td, J = 13.14, 5.40 Hz, 1 H), 1.81-1.98
(m, 4 H), 1.66-1.79 (m, 1 H), 1.44-1.65 (m, 2 H), 1.05 (d, J = 6.85
Hz, 3 H) 97 .sup.1H NMR, (400 MHz, METHANOL-d4) .delta. ppm 8.93
(s, 1 H), 8.46 (d, J = 5.43 Hz, 1 H), 8.37 (dd, J = 8.63, 3.94 Hz,
1 H), 7.99 (t, J = 8.73 Hz, 1 H), 7.55 (d, J = 5.48 Hz, 1 H), 7.29
(d, J = 9.68 Hz, 2 H), 3.56 (s, 3 H), 3.42-3.45 (m, 1 H), 3.05
(ddd, J = 15.75, 11.59, 3.91 Hz, 1 H), 1.80-1.98 (m, 2 H),
1.66-1.78 (m, 1 H), 1.45-1.65 (m, 8 H), 1.05 (d, J = 6.85 Hz, 3 H)
98 .sup.1H NMR, (300 MHz, <cd3od>) .delta. ppm 8.98 (s, 1 H),
8.49 (d, J = 5.57 Hz, 1 H), 8.40 (dd, J = 8.64, 3.96 Hz, 1 H), 8.02
(t, J = 8.64 Hz, 1 H), 7.60 (d, J = 5.57 Hz, 1 H), 7.35 (d, J =
9.67 Hz, 2 H), 3.58 (s, 3 H), 3.46 (s, 1 H), 3.00-3.15 (m, 1 H),
2.35-2.62 (m, 4 H), 2.02-2.21 (m, 1 H), 1.78-1.99 (m, 3 H),
1.45-1.68 (m, 1 H), 1.07 (d, J = 6.74 Hz, 3 H) 99 .sup.1H NMR, (400
MHz, <cd3od>) .delta. ppm 0.97 (d, J = 6.80 Hz, 3 H)
1.36-1.52 (m, 1 H) 1.63-1.73 (m, 1 H) 1.75-1.91 (m, 7 H) 2.83-3.02
(m, 1 H) 3.25 (s, 2 H) 3.51-3.54 (m, 3 H) 3.56-3.63 (m, 1 H) 4.06
(dt, J = 11.21, 2.95 Hz, 4 H) 7.12 (d, J = 9.19 Hz, 2 H) 7.38 (d, J
= 5.33 Hz, 1 H) 7.97 (t, J = 8.73 Hz, 1 H) 8.29-8.41 (m, 2 H) 8.95
(s, 1 H) 100 .sup.1H NMR, (300 MHz, <cd3od>) .delta. ppm 8.97
(s, 1 H), 8.50 (d, J = 5.57 Hz, 1 H), 8.40 (dd, J = 8.64, 3.96 Hz,
1 H), 8.02 (t, J = 8.79 Hz, 1 H), 7.62 (d, J = 5.57 Hz, 1 H), 7.19
(d, J = 9.08 Hz, 2 H), 4.61 (s, 2 H), 3.63 (q, J = 7.03 Hz, 2 H),
3.58 (s, 3 H), 3.43-3.50 (m, 1 H), 3.01-3.16 (m, 1 H), 1.83-1.99
(m, 2 H), 1.68-1.82 (m, 1 H), 1.48-1.67 (m, 2 H), 1.28 (t, J = 7.03
Hz, 3 H), 1.07 (d, J = 6.74 Hz, 3 H) 101 .sup.1H NMR, (300 MHz,
<cd3od>) .delta. ppm 8.97-9.13 (m, 1 H), 8.48-8.60 (m, 1 H),
8.33-8.45 (m, 1 H), 7.94-8.08 (m, 1 H), 7.60-7.72 (m, 1 H),
6.71-6.87 (m, 2 H), 4.92-5.04 (m, 1 H), 4.65-4.79 (m, 1 H),
3.47-3.53 (m, 1 H) 3.60 (s, 3 H), 3.02-3.20 (m, 1 H), 1.86-2.06 (m,
2 H), 1.71-1.85 (m, 1 H), 1.48-1.70 (m, 2 H), 1.39 (d, J = 6.15 Hz,
6 H), 1.10 (d, J = 6.74 Hz, 3 H) 104 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 9.26 (d, J = 5.1 Hz, 1 H), 8.74 (s, 1
H), 8.55 (d, J = 5.5 Hz, 1 H), 8.21 (d, J = 5.1 Hz, 1 H), 7.51-7.70
(m, 2 H), 7.10-7.27 (m, 2 H), 3.80 (br. s., 1 H), 3.60-3.74 (m, 1
H), 3.20-3.28 (m, 1 H), 3.16 (s, 3 H), 2.32 (quin, J = 12.7 Hz, 2
H), 2.04-2.24 (m, 2 H), 1.65-1.84 (m, 1 H), 1.37 (d, J = 7.0 Hz, 3
H) 107 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 1.36 (d, J
= 7.04 Hz, 3 H) 1.75 (dtd, J = 12.98, 8.61, 8.61, 3.99 Hz, 3 H)
2.01-2.23 (m, 4 H) 2.24-2.39 (m, 2 H) 3.16 (s, 3 H) 3.19-3.28 (m, 1
H) 3.54-3.73 (m, 3 H) 3.79 (br. s., 1 H) 3.89-4.04 (m, 2 H)
4.64-4.75 (m, 1 H) 6.83 (d, J = 9.98 Hz, 2 H) 7.61 (d, J = 5.53 Hz,
1 H) 7.96 (t, J = 8.68 Hz, 1 H) 8.34 (dd, J = 8.61, 3.91 Hz, 1 H)
8.53 (d, J = 5.48 Hz, 1 H) 8.72 (s, 1 H) 108 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 1.36 (d, J = 6.99 Hz, 3 H) 1.75 (d, J =
13.50 Hz, 1 H) 1.80-1.91 (m, 1 H) 2.03-2.23 (m, 3 H) 2.23-2.37 (m,
2 H) 2.38-2.62 (m, 4 H) 3.17 (s, 3 H) 3.61-3.73 (m, 1 H) 3.78 (br.
s., 1 H) 7.34 (d, J = 9.59 Hz, 2 H) 7.59 (d, J = 5.48 Hz, 1 H) 8.01
(t, J = 8.66 Hz, 1 H) 8.38 (dd, J = 8.61, 3.91 Hz, 1 H) 8.52 (d, J
= 5.43 Hz, 1 H) 8.73 (s, 1 H) 109 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm 1.15 (dd, J = 6.46, 1.76 Hz, 7 H)
1.22 (d, J = 6.65 Hz, 4 H) 1.60 (d, J = 12.91 Hz, 1 H) 1.86-2.06
(m, 2 H) 2.08-2.35 (m, 3 H) 4.10 (dd, J = 13.89, 6.85 Hz, 4 H)
7.26-7.47 (m, 4 H) 7.77 (dd, J = 8.61, 2.35 Hz, 1 H) 7.88 (ddd, J =
8.31, 4.60, 2.35 Hz, 1 H) 8.15 (d, J = 3.13 Hz, 3 H) 8.35 (d, J =
7.83 Hz, 1 H) 8.40-8.54 (m, 2 H) 8.76 (s, 1 H) 10.36 (s, 1 H) 110
.sup.1H NMR (400 MHz, <dmso-.sub.d6>) .delta. ppm 1.23 (d, J
= 6.65 Hz, 5 H) 1.59 (br. s., 2 H) 1.78-2.37 (m, 14 H) 3.00-3.13
(m, 2 H) 3.51-3.73 (m, 6 H) 3.77 (br. s., 2 H) 3.89 (dd, J = 11.35,
5.09 Hz, 4 H) 7.29 (d, J = 5.48 Hz, 2 H) 7.39-7.55 (m, 4 H)
8.04-8.24 (m, 7 H) 8.32 (dd, J = 8.61, 4.30 Hz, 2 H) 8.44-8.57 (m,
3 H) 10.48-10.57 (m, 2 H) 111 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm 0.95 (d, J = 6.65 Hz, 2 H) 1.15
(d, J = 6.26 Hz, 4 H) 1.48 (br. s., 3 H) 2.75-2.86 (m, 1 H) 3.02
(br. s., 1 H) 3.64-3.81 (m, 2 H) 4.00-4.18 (m, 1 H) 7.24-7.50 (m, 2
H) 7.78 (dd, J = 8.61, 2.35 Hz, 1 H) 7.83-8.01 (m, 3 H) 8.35 (d, J
= 7.43 Hz, 1 H) 8.39-8.49 (m, 1 H) 8.88 (s, 1 H) 10.34 (s, 1 H) 112
.sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 1.36 (d, J = 7.04
Hz, 3 H) 1.71-1.83 (m, 1 H) 2.05-2.25 (m, 2 H) 2.25-2.41 (m, 2 H)
3.13-3.21 (m, 3 H) 3.23-3.29 (m, 1 H) 3.63-3.73 (m, 1 H) 3.80-3.86
(m, 1 H) 3.89 (s, 3 H) 6.80 (d, J = 9.78 Hz, 2 H) 7.59-7.71 (m, 1
H) 7.97 (s, 1 H) 8.31-8.40 (m, 1 H) 8.51-8.59 (m, 1 H) 8.83 (s, 1
H) 114 .sup.1H NMR (400 MHz, <cd3od>) .delta. ppm 1.32 (d, J
= 6.65 Hz, 3 H) 1.69-1.84 (m, 1 H) 2.04-2.19 (m, 2 H) 2.20-2.45 (m,
2 H) 3.13-3.22 (m, 3 H) 3.24-3.29 (m, 1 H) 3.57-3.73 (m, 1 H) 3.89
(br. s., 1 H) 7.05-7.19 (m, 3 H) 7.47-7.59 (m, 1 H) 7.76 (d, J =
5.48 Hz, 1 H) 8.45-8.59 (m, 1 H) 9.16 (s, 1 H) 115 .sup.1H NMR,
(400 MHz, <cd3od>) .delta. ppm 1.26 (d, J = 6.65 Hz, 3 H)
1.54-1.67 (m, 1 H) 1.89-2.02 (m, 2 H) 2.08 (q, J = 12.52 Hz, 1 H)
2.94-3.15 (m, 1H) 3.16-3.24 (m, 3 H) 3.65 (br. s., 1 H) 3.81 (br.
s., 1 H) 7.10-7.34 (m, 3 H) 7.44-7.68 (m, 3 H) 8.15 (d, J = 3.91
Hz, 3 H) 8.46 (d, J = 5.48 Hz, 1 H) 8.62 (s, 1 H) 9.59 (s, 1 H) 116
.sup.1H NMR (400 MHz, <dmso-.sub.d6>) .delta. ppm 1.03-1.31
(m, 6 H) 1.57 (d, J = 12.91 Hz, 1 H) 1.81-2.08 (m, 3 H) 2.09-2.20
(m, 1 H) 2.92-3.11 (m, 1 H) 3.17 (s, 3 H) 3.53 (q, J = 7.04 Hz, 2
H) 3.60 (d, J = 5.87 Hz, 1 H) 3.79 (br. s., 1 H) 4.48-4.67 (m, 2 H)
7.15-7.37 (m, 3 H) 8.01-8.22 (m, 4 H) 8.31 (dd, J = 8.61, 3.91 Hz,
1 H) 8.44-8.59 (m, 2 H) 10.32-10.60 (m, 1 H) 117 .sup.1H NMR, (400
MHz, <cd3od>) .delta. ppm 9.03 (s, 1 H), 8.45 (d, J = 5.1 Hz,
1 H), 8.40 (dd, J = 8.6, 3.9 Hz, 1 H), 8.02 (t, J = 8.6 Hz, 1 H),
7.53 (d, J = 5.5 Hz, 1 H), 7.35 (d, J = 10.2 Hz, 2 H), 3.91-4.03
(m, 2 H), 3.79-3.90 (m, 2 H), 3.53 (s, 3 H), 3.05-3.21 (m, 2 H),
2.93 (t, J = 10.2 Hz, 1 H), 2.09-2.26 (m, 3 H), 1.93 (dd, J = 13.5,
2.5 Hz, 1 H), 1.78 (q, J = 12.1 Hz, 1 H), 1.67 (d, J = 13.7 Hz, 2
H), 1.21-1.51 (m, 2 H), 1.07 (d, J = 6.3 Hz, 3 H) 118 .sup.1H NMR,
(400 MHz, <cd3od>) .delta. ppm 8.95 (s, 1 H), 8.51 (d, J =
5.5 Hz, 1 H), 8.40 (dd, J = 8.6, 3.9 Hz, 1 H), 8.02 (t, J = 8.6 Hz,
1 H), 7.63 (d, J = 5.5 Hz, 1 H), 7.35 (d, J = 9.8 Hz, 2 H), 4.26
(dt, J = 10.7, 5.0 Hz, 1 H), 4.00-4.12 (m, 1 H), 3.91-4.00 (m, 2
H), 3.79-3.90 (m, 2 H), 3.69 (s, 1 H), 3.55 (t, J = 5.3 Hz, 2 H),
3.35-3.46 (m, 1 H), 3.03-3.20 (m, 4 H), 2.16 (td, J = 13.0, 4.9 Hz,
2 H), 1.88-2.01 (m, 2 H), 1.78 (d, J = 6.3 Hz, 1 H), 1.49-1.72 (m,
4 H), 1.11 (d, J = 6.7 Hz, 3 H) 119 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 8.76 (s, 1 H), 8.52 (d, J = 5.5 Hz, 1
H), 8.34 (dd, J = 8.6, 3.9 Hz, 1 H), 7.97 (t, J = 8.8 Hz, 1 H),
7.61 (d, J = 5.5 Hz, 1 H), 6.82 (d, J = 10.2 Hz, 2 H), 4.21 (dd, J
= 5.1, 3.5 Hz, 2 H), 3.73-3.87 (m, 3 H), 3.63-3.72 (m, 1 H), 3.43
(s, 3 H), 3.19-3.28 (m, 1 H), 3.17 (s, 3 H), 2.32 (qd, J = 12.7,
6.1 Hz, 2 H), 2.05-2.24 (m, 2 H), 1.75 (d, J = 13.3 Hz, 1 H), 1.36
(d, J = 7.0 Hz, 3 H) 120 .sup.1H NMR, (300 MHz, <cd3od>)
.delta. ppm 8.77 (s, 1 H), 8.62 (s, 1 H), 8.54 (d, J = 5.5 Hz, 1
H), 7.55-7.69 (m, 2 H), 7.23 (t, J = 8.6 Hz, 2 H), 3.81 (br. s., 1
H), 3.67-3.78 (m, 1 H), 3.17 (s, 3 H), 2.20-2.46 (m, 3 H),
2.08-2.19 (m, 1 H), 1.78 (d, J = 13.3 Hz, 1 H), 1.40 (d, J = 7.0
Hz, 3 H) 121 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 8.72
(s, 1 H), 8.53 (d, J = 5.5 Hz, 1 H), 8.38 (dd, J = 8.6, 3.9 Hz, 1
H), 8.00 (t, J = 8.6 Hz, 1 H), 7.62 (d, J = 5.5 Hz, 1 H), 7.34 (d,
J = 9.8 Hz, 2 H), 3.82-4.01 (m, 4 H), 3.79 (br. s., 1 H), 3.63-3.73
(m, 1 H), 3.20-3.28 (m, 1 H), 3.17 (s, 3 H), 2.31 (qd, J = 12.7,
8.4 Hz, 2 H), 2.06-2.24 (m, 4 H), 1.75 (d, J = 13.3 Hz, 1 H), 1.65
(d, J = 13.3 Hz, 2 H), 1.36 (d, J = 7.0 Hz, 3 H) 122 .sup.1H NMR,
(400 MHz, <cd3od>) .delta. ppm 8.77 (s, 1 H), 8.54 (d, J =
5.5 Hz, 1 H), 8.36 (dd, J = 8.8, 4.1 Hz, 1 H), 7.99 (t, J = 8.6 Hz,
1 H), 7.63 (d, J = 5.5 Hz, 1 H), 7.03 (d, J = 9.4 Hz, 2 H), 3.80
(br. s., 1 H), 3.62-3.73 (m, 1 H), 3.20-3.29 (m, 1 H), 3.17 (s, 3
H), 2.46 (s, 3 H), 2.25-2.40 (m, 2 H), 2.06-2.24 (m, 2 H),
1.68-1.82 (m, 1 H), 1.36 (d, J = 7.0 Hz, 3 H) 123 .sup.1H NMR, (400
MHz, <cd3od>) .delta. ppm 0.90 (d, J = 6.65 Hz, 3 H)
1.72-1.81 (m, 2 H) 1.88-2.07 (m, 2 H) 2.07-2.21 (m, 1 H) 3.24-3.28
(m, 1 H) 3.53-3.63 (m, 1 H) 4.87-4.97 (m, 1 H) 5.59-5.69 (m, 1 H)
6.94-7.08 (m, 2 H) 7.23 (s, 2 H) 7.58-7.70 (m, 1 H) 7.72-7.84 (m, 2
H) 8.01-8.09 (m, 1 H) 8.15-8.22 (m, 1 H) 8.39-8.48 (m, 1 H)
8.55-8.62 (m, 1 H) 9.08 (s, 1 H) 124 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 1.08 (d, J = 6.26 Hz, 3 H) 1.43 (q, J =
12.52 Hz, 1 H) 1.59-1.75 (m, 1 H) 1.81 (q, J = 12.39 Hz, 1 H) 1.94
(dd, J = 13.30, 2.74 Hz, 1 H) 2.15-2.30 (m, 1 H) 2.96 (t, J = 9.98
Hz, 1 H) 3.08-3.22 (m, 2 H) 3.44 (s, 3 H) 3.53 (s, 3 H) 3.73-3.83
(m, 2 H) 4.11-4.28 (m, 2 H) 6.83 (d, J = 10.17 Hz, 2 H) 7.69 (d, J
= 5.48 Hz, 1 H) 8.00 (t, J = 8.80 Hz, 1 H) 8.38 (dd, J = 8.61, 3.52
Hz, 1 H) 8.51 (d, J = 5.48 Hz, 1 H) 9.20 (s, 1 H) 125 .sup.1H NMR
(400 MHz, <dmso-.sub.d6>) .delta. ppm 1.03 (d, J = 6.65 Hz, 3
H) 1.33 (q, J = 12.52 Hz, 1 H) 1.45-1.68 (m, 2 H) 1.70-1.88 (m, 2
H) 2.86-3.04 (m, 1H) 3.04-3.15 (m, 3 H) 3.22-3.43 (m, 3 H) 3.58
(br. s., 2 H) 3.61-3.72 (m, 2 H) 3.89 (dt, J = 10.08, 4.94 Hz, 6 H)
4.02 (td, J = 9.20, 4.30 Hz, 5 H)7.15-7.34 (m, 3 H) 7.45-7.69 (m, 3
H) 7.95 (br. s., 3 H) 8.42 (d, J = 5.48 Hz, 1 H) 8.78 (s, 1 H) 9.48
(s, 1 H) 126 .sup.1H NMR (400 MHz, <dmso-.sub.d6>) .delta.
ppm 0.93 (d, J = 6.26 Hz, 7 H) 1.15 (d, J = 6.65 Hz, 14 H)
1.32-1.46 (m, 2 H) 1.49-1.70 (m, 5 H) 1.72-1.85 (m, 2 H) 2.09 (d, J
= 10.17 Hz, 2 H) 2.88 (t, J = 9.98 Hz, 2 H) 2.98-3.14 (m, 5 H) 3.30
(s, 7 H) 4.00-4.17 (m, 2 H) 7.31-7.41 (m, 4 H) 7.45 (d, J = 5.09
Hz, 2 H) 7.79 (dd, J = 9.00, 1.96 Hz, 2 H) 7.85-8.00 (m, 8 H) 8.33
(d, J = 7.83 Hz, 2 H) 8.46 (d, J = 5.09 Hz, 4 H) 8.89 (s, 2 H)
10.36 (s, 2 H) 127 .sup.1H NMR (400 MHz, <dmso-.sub.d6>)
.delta. ppm 0.95 (d, J = 6.26 Hz, 3 H) 1.19-1.39 (m, 1 H) 1.47-1.67
(m, 2 H) 1.74 (d, J = 11.35 Hz, 1 H) 1.87 (t, J = 11.93 Hz, 2 H)
2.00 (br. s., 1 H) 2.11-2.37 (m, 2 H) 2.82-3.13 (m, 3 H) 3.37 (s, 3
H) 3.69 (t, J = 11.35 Hz, 2 H) 3.89 (dd, J = 11.35, 5.09 Hz, 2 H)
7.37 (d, J = 5.48 Hz, 1 H) 7.44 (d, J = 9.39 Hz, 2 H) 7.91 (br. s.,
3 H) 8.15-8.25 (m, 1 H) 8.33
(dd, J = 8.80, 4.11 Hz, 1 H) 8.44 (d, J = 5.09 Hz, 1 H) 8.62-8.70
(m, 1 H) 10.39 (s, 1 H) 128 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm 0.96 (d, J = 6.65 Hz, 3 H) 1.16
(d, J = 6.65 Hz, 6 H) 1.29-1.64 (m, 3 H) 1.70-1.88 (m, 2 H)
2.97-3.14 (m, 4 H) 3.30-3.44 (m, 2 H) 3.53 (br. s., 1 H) 3.57-3.69
(m, 1 H) 3.88 (dt, J = 10.17, 5.09 Hz, 1 H) 4.00 (td, J = 9.19,
4.30 Hz, 1 H) 4.11 (dq, J = 13.60, 6.68 Hz, 1 H) 7.25-7.47 (m, 3 H)
7.78 (dd, J = 8.80, 2.15 Hz, 1 H) 7.84-8.01 (m, 4 H) 8.28-8.50 (m,
3 H) 8.90 (s, 1 H) 10.35 (s, 1 H) 129 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm 0.98 (d, J = 7.04 Hz, 3 H) 1.30
(q, J = 12.52 Hz, 1 H) 1.47 (d, J = 12.52 Hz, 1 H) 1.53-1.79 (m, 3
H) 1.87 (t, J = 11.93 Hz, 2 H) 2.10-2.38 (m, 2 H) 2.91 (t, J =
12.33 Hz, 1 H) 3.09 (s, 3 H) 3.26 (br. s., 1 H) 3.32-3.44 (m, 1 H)
3.52 (br. s., 1 H) 3.58-3.77 (m, 3 H) 3.79-3.95 (m, 3 H) 4.01 (td,
J = 9.19, 4.30 Hz, 1 H) 7.23 (d, J = 5.09 Hz, 1 H) 7.38-7.53 (m, 2
H) 7.87 (br. s., 3 H) 8.19 (t, J = 8.80 Hz, 1 H) 8.32 (dd, J =
8.61, 3.91 Hz, 1 H) 8.45 (d, J = 5.09 Hz, 1 H) 8.53-8.63 (m, 1 H)
10.37-10.49 (m, 1 H) 130 .sup.1H NMR, (400 MHz, <cd3od>)
.delta. ppm 1.13 (d, J = 6.65 Hz, 3 H) 1.46-1.72 (m, 2 H) 1.79 (d,
J = 5.87 Hz, 1 H) 1.88-2.05 (m, 2 H) 3.10 (s, 4 H) 3.35-3.47 (m, 4
H) 3.50-3.63 (m, 2 H) 3.71 (s, 1 H) 3.76-3.81 (m, 2 H) 4.05 (ddd, J
= 10.86, 6.95, 4.11 Hz, 1 H) 4.17-4.33 (m, 3 H) 6.83 (d, J = 10.17
Hz, 2 H) 7.71 (d, J = 5.48 Hz, 1 H) 7.99 (t, J = 8.61 Hz, 1 H) 8.37
(dd, J = 8.80, 3.72 Hz, 1 H) 8.54 (d, J = 5.87 Hz, 1 H) 9.09 (s, 1
H) 131 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.53 (d, J
= 7.04 Hz, 3 H) 2.57-2.82 (m, 1 H) 3.56-3.72 (m, 1 H) 3.83 (dd, J =
13.30, 3.91 Hz, 1 H) 4.04 (d, J = 10.17 Hz, 1 H) 4.12-4.36 (m, 2 H)
5.20 (t, J = 3.91 Hz, 1 H) 7.16-7.36 (m, 2 H) 7.50-7.71 (m, 2 H)
7.89 (d, J = 0.78 Hz, 1 H) 8.12 (d, J = 0.78 Hz, 1 H) 8.34-8.48 (m,
1 H) 8.73 (s, 1 H) 9.08 (s, 1 H) 132 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 0.34 (d, J = 6.65 Hz, 3 H) 2.22-2.53 (m,
1 H) 3.36 (s, 3 H) 3.39-3.56 (m, 1 H) 3.60-3.76 (m, 3 H) 3.85 (d, J
= 6.65 Hz, 1 H) 3.93-4.03 (m, 1 H) 4.10-4.25 (m, 3 H) 4.94 (br. s.,
1 H) 6.80 (d, J = 10.17 Hz, 2 H) 7.46 (d, J = 7.04 Hz, 1 H) 7.81
(s, 1 H) 7.92 (t, J = 8.61 Hz, 1 H) 8.00 (d, J = 0.78 Hz, 1 H)
8.26-8.45 (m, 2 H) 9.08 (s, 1 H) 133 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 1.19 (d, J = 6.65 Hz, 3 H) 1.56-1.77 (m,
2 H) 1.84-2.05 (m, 3 H) 3.11 (s, 3 H) 3.17-3.27 (m, 1 H) 3.47 (ddd,
J = 11.64, 4.40, 2.74 Hz, 1 H) 3.53-3.63 (m, 2 H) 3.73 (s, 1 H)
4.06 (ddd, J = 10.76, 6.46, 4.70 Hz, 1 H) 4.23-4.37 (m, 1 H) 7.24
(t, J = 8.61 Hz, 2 H) 7.62 (tt, J = 8.61, 6.26 Hz, 1 H) 7.73 (d, J
= 5.87 Hz, 1 H) 8.55 (d, J = 5.48 Hz, 1 H) 8.66 (s, 1 H) 9.02 (s, 1
H) 134 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 1.15 (d, J
= 6.26 Hz, 3 H) 1.43-1.58 (m, 1 H) 1.69-1.92 (m, 2 H) 1.93-2.08 (m,
1 H) 2.20-2.35 (m, 1 H) 3.00 (t, J = 10.17 Hz, 1 H) 3.17-3.29 (m, 2
H) 3.54 (s, 3 H) 7.25 (t, J = 8.61 Hz, 2 H) 7.62 (tt, J = 8.56,
6.31 Hz, 1 H) 7.76 (d, J = 5.87 Hz, 1 H) 8.55 (d, J = 5.48 Hz, 1 H)
8.67 (s, 1 H) 9.11 (s, 1 H) 135 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm 1.03 (d, J = 6.65 Hz, 3 H) 1.33
(q, J = 12.52 Hz, 1 H) 1.45-1.68 (m, 2 H) 1.70-1.88 (m, 2 H)
2.86-3.04 (m, 1H) 3.04-3.15 (m, 3 H) 3.22-3.43 (m, 3 H) 3.58 (br.
s., 2 H) 3.61-3.72 (m, 2 H) 3.89 (dt, J = 10.08, 4.94 Hz, 6 H) 4.02
(td, J = 9.20, 4.30 Hz, 5 H) 7.15-7.34 (m, 3 H) 7.45-7.69 (m, 3 H)
7.95 (br. s., 3 H) 8.42 (d, J = 5.48 Hz, 1 H) 8.78 (s, 1 H) 9.48
(s, 1 H) 136 1H NMR (400 MHz, <dmso>) d ppm 0.83-1.01 (m, 3
H) 1.09-1.22 (m, 3 H) 1.30 (q, J = 12.52 Hz, 1 H) 1.45-1.68 (m, 2
H) 1.74 (d, J = 10.96 Hz, 1H) 2.02 (d, J = 9.78 Hz, 1 H) 2.82-2.95
(m, 1 H) 2.95-3.12 (m, 2 H) 3.37 (s, 3 H) 3.54 (q, J = 7.04 Hz, 2
H) 4.57 (s, 2 H) 7.09-7.32 (m, 2 H) 7.43 (d, J = 5.09 Hz, 1 H) 7.99
(br. s., 3 H) 8.10-8.23 (m, 1 H) 8.32 (dd, J = 8.61, 4.30 Hz, 1 H)
8.47 (d, J = 5.09 Hz, 1 H) 8.71 (s, 1 H) 10.41 (s, 1 H) 137 .sup.1H
NMR, (400 MHz, <cd3od>) .delta. ppm 1.09 (d, J = 6.46 Hz, 3
H) 1.36-1.50 (m, 1 H) 1.63-1.87 (m, 4 H) 1.95 (dd, J = 13.55, 2.59
Hz, 1 H) 2.05-2.14 (m, 2 H) 2.17-2.29 (m, 1 H) 2.91-3.01 (m, 1 H)
3.10-3.23 (m, 2 H) 3.54 (s, 3 H) 3.64 (ddd, J = 11.73, 8.74, 3.01
Hz, 2 H) 3.93-4.02 (m, 2 H) 4.66-4.75 (m, 1 H) 6.85 (d, J = 10.12
Hz, 2 H) 7.64 (d, J = 5.53 Hz, 1 H) 8.00 (t, J = 8.71 Hz, 1 H) 8.38
(dd, J = 8.63, 3.89 Hz, 1 H) 8.50 (d, J = 5.48 Hz, 1 H) 9.12 (s, 1
H) 138 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 1.09 (d, J
= 6.50 Hz, 3 H) 1.36-1.51 (m, 1 H) 1.64-2.00 (m, 4 H) 2.04-2.27 (m,
2 H) 2.38-2.50 (m, 2 H) 2.51-2.63 (m, 2 H) 2.85-3.03 (m, 2 H)
3.11-3.24 (m, 2 H) 3.54 (s, 3 H) 7.36 (d, J = 9.49 Hz, 2 H) 7.64
(d, J = 5.58 Hz, 1 H) 8.04 (t, J = 8.71 Hz, 1 H) 8.42 (dd, J =
8.66, 3.91 Hz, 1 H) 8.50 (d, J = 5.58 Hz, 1 H) 9.15 (s, 1 H) 139
.sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 1.12 (d, J = 6.85
Hz, 3 H) 1.52-1.71 (m, 2 H) 1.72-1.98 (m, 4 H) 2.05-2.18 (m, 1 H)
2.35-2.60 (m, 4 H) 3.06-3.17 (m, 4 H) 3.55 (t, J = 5.31 Hz, 2 H)
3.68 (s, 1 H) 3.97-4.11 (m, 1 H) 4.19-4.34 (m, 1 H) 7.35 (d, J =
9.44 Hz, 2 H) 7.58 (br. s., 1 H) 8.02 (t, J = 8.71 Hz, 1 H) 8.40
(dd, J = 8.73, 3.94 Hz, 1 H) 8.49 (d, J = 5.38 Hz, 1 H) 8.91 (br.
s., 1 H) 140 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 1.14
(d, J = 6.94 Hz, 3 H) 1.55-1.83 (m, 5 H) 1.87-2.01 (m, 2 H)
2.03-2.18 (m, 2 H) 3.12 (s, 3 H) 3.14 (dd, J = 3.28, 1.66 Hz, 1 H)
3.52-3.73 (m, 5 H) 3.91-4.11 (m, 3 H) 4.22-4.35 (m, 1 H) 4.72 (tt,
J = 8.08, 4.13 Hz, 1 H) 6.86 (d, J = 10.08 Hz, 2 H) 7.64 (d, J =
5.53 Hz, 1 H) 7.99 (t, J = 8.71 Hz, 1 H) 8.37 (dd, J = 8.63, 3.94
Hz, 1 H) 8.52 (d, J = 5.53 Hz, 1 H) 8.95 (s, 1 H) 141 .sup.1H NMR,
(400 MHz, <cd3od>) .delta. ppm 1.08 (d, J = 6.50 Hz, 3 H)
1.34-1.49 (m, 7 H) 1.63-1.76 (m, 1 H) 1.81-1.99 (m, 2 H) 2.18-2.28
(m, 1 H) 2.99 (t, J = 10.00 Hz, 1 H) 3.12-3.25 (m, 2 H) 3.54 (s, 3
H) 4.72 (dt, J = 12.07, 6.02 Hz, 1 H) 6.76 (d, J = 10.22 Hz, 2 H)
7.75 (d, J = 5.62 Hz, 1 H) 7.99 (t, J = 8.73 Hz, 1 H) 8.38 (dd, J =
8.63, 3.89 Hz, 1 H) 8.53 (d, J = 5.48 Hz, 1 H) 9.26 (s, 1 H) 142
.sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 1.12 (d, J = 6.85
Hz, 3 H) 1.38 (d, J = 6.02 Hz, 6 H) 1.56-1.64 (m, 1 H) 1.65-1.73
(m, 1 H) 1.76-1.86 (m, 1 H) 1.92-2.01 (m, 2 H) 3.11 (s, 3 H)
3.13-3.22 (m, 1 H) 3.37-3.45 (m, 1 H) 3.47-3.64 (m, 2 H) 3.74 (s, 1
H) 4.06 (ddd, J = 10.69, 7.31, 3.72 Hz, 1 H) 4.27 (ddd, J = 10.45,
6.39, 3.94 Hz, 1 H) 4.72 (dt, J = 12.07, 6.02 Hz, 1 H) 6.78 (d, J =
10.17 Hz, 2 H) 7.74 (d, J = 5.72 Hz, 1 H) 7.99 (t, J = 8.71 Hz, 1
H) 8.37 (dd, J = 8.61, 3.91 Hz, 1 H) 8.55 (d, J = 5.62 Hz, 1 H)
9.13 (s, 1 H) 143 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm
1.08 (d, J = 6.26 Hz, 3 H) 1.34-1.53 (m, 1 H) 1.60-1.75 (m, 1 H)
1.79-2.00 (m, 2 H) 2.17-2.29 (m, 1 H) 2.98 (s, 1 H) 3.09-3.24 (m, 2
H) 3.48-3.58 (m, 3 H) 3.91 (s, 3 H) 6.80 (d, J = 9.78 Hz, 2 H) 7.77
(s, 1 H) 8.00 (s, 1 H) 8.31-8.44 (m, 1 H) 8.47-8.60 (m, 1 H) 9.28
(s, 1 H) 144 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 1.13
(d, J = 7.04 Hz, 3 H) 1.51-1.72 (m, 2 H) 1.74-1.87 (m, 1 H)
1.89-1.98 (m, 2 H) 3.11 (s, 4 H) 3.35-3.43 (m, 1 H) 3.51-3.60 (m, 2
H) 3.66-3.74 (m, 1 H) 3.86-3.95 (m, 3 H) 4.00-4.11 (m, 1 H)
4.22-4.32 (m, 1 H) 6.75-6.87 (m, 2 H) 7.64-7.72 (m, 1 H) 7.93-8.03
(m, 1 H) 8.33-8.42 (m, 1 H) 8.48-8.57 (m, 1 H) 9.00-9.08 (m, 1 H)
145 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 1.03-1.11 (m,
3 H) 1.34-1.49 (m, 1 H) 1.63 (ddd, J = 12.42, 6.16, 3.33 Hz, 1 H)
1.81 (q, J = 12.13 Hz, 1 H) 1.92 (dd, J = 13.30, 2.74 Hz, 1 H)
2.14-2.27 (m, 1 H) 2.89-2.99 (m, 1 H) 3.05-3.19 (m, 2 H) 3.53 (s, 3
H) 7.10-7.18 (m, 3 H) 7.49-7.59 (m, 1 H) 7.70 (d, J = 5.48 Hz, 1 H)
8.47 (d, J = 5.48 Hz, 1 H) 9.35 (s, 1 H) 146 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm 0.96 (d, J = 6.65 Hz, 3 H) 1.26
(d, J = 12.52 Hz, 1 H) 1.46 (br. s., 1 H) 1.54-1.80 (m, 3 H) 2.90
(br. s., 1 H) 3.07 (s, 3 H) 3.15-3.27 (m, 1 H) 3.38 (br. s., 1 H)
3.55 (br. s., 1 H) 3.64 (d, J = 5.48 Hz, 1 H) 3.87-3.91 (m, 1 H)
4.00 (d, J = 3.91 Hz, 1 H) 7.20-7.29 (m, 4 H) 7.30-7.44 (m, 1 H)
7.58 (s, 1 H) 7.95 (br. s., 2 H) 8.43 (d, J = 5.09 Hz, 1 H) 8.78
(s, 1 H) 10.12 (s, 1 H) 147 .sup.1H NMR, (400 MHz, <cd3od>)
.delta. ppm 0.44 (d, J = 6.85 Hz, 3 H) 1.40 (dd, J = 6.02, 3.03 Hz,
6 H) 2.54 (ddd, J = 11.57, 4.62, 2.20 Hz, 1 H) 3.52-3.62 (m, 1 H)
3.63-3.72 (m, 1 H) 3.88 (dd, J = 11.64, 4.16 Hz, 1 H) 4.03-4.11 (m,
1 H) 4.14-4.23 (m, 1 H) 4.78 (dt, J = 12.06, 6.05 Hz, 1 H) 5.09 (t,
J = 4.35 Hz, 1 H) 6.84 (d, J = 10.42 Hz, 2 H) 7.52 (d, J = 6.50 Hz,
1 H) 7.89 (d, J = 1.03 Hz, 1 H) 8.00 (t, J = 8.71 Hz, 1 H) 8.09 (d,
J = 1.03 Hz, 1 H) 8.34-8.46 (m, 2 H) 9.18 (s, 1 H) 148 .sup.1H NMR,
(400 MHz, <cd3od>) .delta. ppm 0.65 (d, J = 6.85 Hz, 3 H)
1.40 (dd, J = 6.02, 2.93 Hz, 6 H) 2.17-2.27 (m, 1 H) 2.83 (t, J =
12.98 Hz, 1 H) 3.22-3.29 (m, 1 H) 3.47-3.62 (m, 2 H) 3.64-3.73 (m,
4 H) 4.06-4.15 (m, 1 H) 4.76 (dt, J = 12.07, 6.02 Hz, 1 H) 6.83 (d,
J = 10.51 Hz, 2 H) 7.25 (d, J = 9.68 Hz, 1 H) 7.51 (d, J = 6.60 Hz,
1 H) 8.01 (t, J = 8.66 Hz, 1 H) 8.37-8.44 (m, 2 H) 9.33 (s, 1 H)
149 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.94 (d, J =
6.75 Hz, 3 H) 1.38 (d, J = 6.02 Hz, 6 H) 1.43-1.57 (m, 1 H) 1.71
(d, J = 13.60 Hz, 1 H) 1.78-2.02 (m, 3 H) 3.10-3.21 (m, 1 H) 3.41
(dt, J = 12.41, 3.58 Hz, 1 H) 3.72 (s, 3 H) 4.08-4.16 (m, 1 H) 4.72
(dt, J = 12.09, 6.06 Hz, 1 H) 6.78 (d, J = 10.22 Hz, 2 H) 7.21 (d,
J = 9.73 Hz, 1 H) 7.82 (d, J = 5.62 Hz, 1 H) 7.98 (t, J = 8.71 Hz,
1 H) 8.37 (dd, J = 8.61, 3.91 Hz, 1 H) 8.54 (d, J = 5.58 Hz, 1 H)
9.08 (s, 1 H) 150 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm
0.91-1.01 (m, 3 H) 1.73-1.86 (m, 1 H) 1.90-2.01 (m, 1 H) 2.12-2.31
(m, 2 H) 2.34-2.46 (m, 1 H) 3.53-3.66 (m, 1 H) 3.77-3.88 (m, 1 H)
5.15-5.25 (m, 1 H) 6.03-6.13 (m, 1 H) 6.52-6.61 (m, 1 H) 7.10-7.24
(m, 2 H) 7.42-7.50 (m, 1 H) 7.53-7.66 (m, 1 H) 7.70-7.77 (m, 1 H)
7.77-7.85 (m, 1 H) 7.99-8.05 (m, 1 H) 8.36-8.43 (m, 1 H) 8.51-8.58
(m, 1 H) 8.76-8.82 (m, 1 H) 151 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm -0.10-0.12 (m, 3 H) 1.64 (br. s.,
2 H) 1.85-2.12 (m, 3 H) 3.43 (t, J = 11.15 Hz, 4 H) 3.52-3.73 (m, 5
H) 4.68 (br. s., 1 H) 7.07-7.26 (m, 3 H) 7.61 (s, 1 H) 7.85 (s, 3
H) 7.92-8.00 (m, 1 H) 8.05-8.18 (m, 2 H) 8.59 (br. s., 1 H) 10.16
(br. s., 1 H) 152 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm
0.95 (d, J = 7.04 Hz, 3 H) 1.72-1.87 (m, 1 H) 1.96 (d, J = 14.09
Hz, 1 H) 2.11-2.46 (m, 3 H) 3.60 (br. s., 1 H) 3.84 (br. s., 1 H)
5.20 (d, J = 5.48 Hz, 1 H) 6.09 (t, J = 6.46 Hz, 1 H) 6.57 (d, J =
9.00 Hz, 1 H) 7.16 (t, J = 8.61 Hz, 2 H) 7.46 (t, J = 7.83 Hz, 1 H)
7.59 (quin, J = 7.43 Hz, 1 H) 7.74 (d, J = 7.04 Hz, 1 H) 7.86 (br.
s., 1 H) 8.03 (t, J = 8.80 Hz, 1 H) 8.40 (dd, J = 8.41, 3.72 Hz, 1
H) 8.55 (d, J = 5.09 Hz, 1 H) 8.83 (d, J = 9.00 Hz, 1 H) 153
.sup.1H NMR (400 MHz, <dmso-.sub.d6>) .delta. ppm 0.96 (d, J
= 6.65 Hz, 3 H) 1.25-1.40 (m, 1 H) 1.40-1.52 (m, 1 H) 1.56-1.76 (m,
3 H) 1.77-1.93 (m, 2 H) 2.11-2.36 (m, 2 H) 2.74-2.99 (m, 3 H) 3.24
(br. s., 1 H) 3.83-3.93 (m, 2 H) 7.24 (d, J = 5.09 Hz, 1 H)
7.36-7.50 (m, 2 H) 7.86 (br. s., 3 H) 8.18 (t, J = 9.00
Hz, 1 H) 8.30 (dd, J = 8.80, 4.11 Hz, 1 H) 8.45 (d, J = 5.09 Hz, 1
H) 8.56 (s, 1 H) 10.41 (s, 1 H) 154 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm 0.52 (d, J = 6.65 Hz, 3 H) 1.64
(d, J = 13.30 Hz, 1 H) 1.79-2.02 (m, 2 H) 2.16-2.41 (m, 2 H) 3.20
(t, J = 12.13 Hz, 1 H) 3.81 (br. s., 1 H) 4.91 (t, J = 3.72 Hz, 1
H) 7.26 (t, J = 8.41 Hz, 2 H) 7.47-7.68 (m, 4 H) 7.80-7.87 (m, 1 H)
8.00 (br. s., 3 H) 8.13 (s, 1 H) 8.51 (d, J = 5.48 Hz, 1 H) 8.72
(s, 1 H) 9.60 (s, 1 H) 155 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm 0.48 (d, J = 6.65 Hz, 3 H)
1.07-1.24 (m, 3 H) 1.60 (d, J = 13.30 Hz, 1 H) 1.78-1.97 (m, 2 H)
2.10-2.23 (m, 1H) 2.33 (q, J = 12.52 Hz, 1 H) 3.15 (t, J = 12.33
Hz, 1 H) 3.52 (q, J = 7.04 Hz, 2 H) 4.49-4.62 (m, 2 H) 4.88 (t, J =
3.72 Hz, 1 H) 7.27 (d, J = 8.61 Hz, 2H) 7.52 (d, J = 5.48 Hz, 1 H)
7.79-7.87 (m, 1 H) 8.00 (br. s., 3 H) 8.17 (t, J = 8.80 Hz, 1 H)
8.31 (dd, J = 8.61, 3.91 Hz, 1 H) 8.55 (d, J = 5.09 Hz, 1 H), 8.58
(s, 1 H) 10.52 (s, 1 H) 156 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm 0.01 (d, J = 7.04 Hz, 3 H)
2.94-3.13 (m, 2 H) 3.21 (d, J = 10.56 Hz, 2 H) 3.31-3.44 (m, 1 H)
3.45-3.63 (m, 2H) 4.71 (t, J = 3.91 Hz, 1 H) 6.89 (t, J = 8.80 Hz,
2 H) 7.06 (d, J = 6.65 Hz, 1 H) 7.12-7.23 (m, 1 H) 7.31 (br. s., 2
H) 7.48 (s, 1 H) 7.80 (s, 2 H) 8.00 (d, J = 6.26 Hz, 1 H) 8.60 (s,
1 H) 9.04 (br. s., 1 H) 157 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm 0.01 (d, J = 7.04 Hz, 3 H)
0.85-0.99 (m, 3 H) 2.01-2.15 (m, 1 H) 3.07-3.24 (m, 1 H) 3.30 (q, J
= 7.04 Hz, 3H) 3.43 (d, J = 10.17 Hz, 2 H) 3.52-3.70 (m, 4 H)
4.26-4.40 (m, 2 H) 4.71 (br. s., 1 H) 6.95-7.09 (m, 2 H) 7.17 (d, J
= 6.65 Hz, 1 H) 7.62 (s, 1 H), 7.88 (s, 1 H) 7.91-8.03 (m, 3 H)
8.07-8.20 (m, 2 H) 8.66 (s, 1 H) 10.17 (br. s., 1 H) 158 .sup.1H
NMR, (400 MHz, <cd3od>) .delta. ppm 0.63 (d, J = 6.85 Hz, 3
H) 1.89 (dt, J = 11.38, 8.98 Hz, 1 H) 2.08-2.22 (m, 2 H) 2.40-2.53
(m, 2 H) 2.54-2.65 (m, 2 H) 2.78 (t, J = 12.76 Hz, 1 H) 3.08-3.27
(m, 2 H) 3.34-3.43 (m, 2 H) 3.46-3.59 (m, 2 H) 3.71 (s, 3 H) 4.02
(br. s., 1 H) 7.38-7.47 (m, 3 H) 8.06 (t, J = 8.68 Hz, 1 H) 8.39
(d, J = 6.46 Hz, 1 H) 8.46 (dd, J = 8.66, 3.96 Hz, 1 H) 9.39 (s, 1
H) 159 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.65 (d, J
= 6.80 Hz, 3 H) 1.74-1.89 (m, 2 H) 2.06-2.25 (m, 3 H) 2.66 (s, 1 H)
2.76 (t, J = 12.18 Hz, 1 H) 3.12-3.22 (m, 1 H) 3.25-3.28 (m, 1 H)
3.36 (dd, J = 3.86, 2.20 Hz, 3 H) 3.45-3.58 (m, 4 H) 3.61-3.70 (m,
3 H) 3.72 (s, 3 H) 3.93-4.04 (m, 2 H) 4.04-4.10 (m, 1 H) 6.88-6.99
(m, 2 H) 7.42 (d, J = 6.31 Hz, 1 H) 8.03 (t, J = 8.68 Hz, 1 H)
8.33-8.47 (m, 2 H) 9.44 (s, 1 H) 160 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 9.16 (s, 1 H), 8.48 (dd, J = 8.6, 3.9
Hz, 1 H), 8.41 (d, J = 6.3 Hz, 1 H), 8.02-8.13 (m, 1 H), 7.93 (s, 1
H), 7.57 (d, J = 6.7 Hz, 1 H), 7.44 (d, J = 9.8 Hz, 2 H), 5.08 (t,
J = 4.1 Hz, 1 H), 4.20-4.32 (m, 1 H), 4.06-4.16 (m, 1 H), 3.95-4.05
(m, 3 H), 3.87-3.95 (m, 2 H), 3.76-3.86 (m, 1 H), 3.56-3.69 (m, 1
H), 2.46-2.60 (m, 1 H), 2.16-2.32 (m, 2 H), 1.73 (d, J = 13.7 Hz, 2
H), 0.43 (d, J = 7.0 Hz, 3 H) 161 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 9.15 (s, 1 H), 8.46 (dd, J = 8.6, 3.9
Hz, 1 H), 8.41 (d, J = 6.7 Hz, 1 H), 8.12 (s, 1 H), 8.05 (t, J =
8.8 Hz, 1 H), 7.92 (s, 1 H), 7.58 (d, J = 7.0 Hz, 1 H), 7.11 (d, J
= 9.8 Hz, 2 H), 5.08 (t, J = 4.3 Hz, 1 H), 4.19-4.31 (m, 1 H),
4.04-4.15 (m, 1 H), 3.99 (dd, J = 11.9, 4.1 Hz, 1 H), 3.80 (dd, J =
13.5, 4.1 Hz, 1 H), 3.60 (t, J = 12.9 Hz, 1 H), 2.45-2.60 (m, 4 H),
0.44 (d, J = 7.0 Hz, 3 H) 162 .sup.1H NMR, (400 MHz, <cd3od>)
.delta. ppm 0.58 (d, J = 6.70 Hz, 3 H) 2.19 (br. s., 1 H) 2.79 (t,
J = 13.01 Hz, 1 H) 3.20-3.28 (m, 1 H) 3.45 (d, J = 11.25 Hz, 1 H)
3.52-3.65 (m, 2 H) 3.71 (s, 3 H) 4.02 (br. s., 1 H) 4.43 (quin, J =
7.25 Hz, 1 H) 4.79 (t, J = 6.31 Hz, 1 H) 5.15-5.22 (m, 2 H) 7.39
(d, J = 9.63 Hz, 2 H) 7.48 (d, J = 6.46 Hz, 1 H) 8.07 (t, J = 8.71
Hz, 1 H) 8.40 (d, J = 6.26 Hz, 1 H) 8.47 (dd, J = 8.66, 3.86 Hz, 1
H) 9.39 (s, 1 H) 163 .sup.1H NMR, (400 MHz, <cd3od>) .delta.
ppm 0.87-0.99 (m, 1 H) 1.14 (d, J = 6.70 Hz, 3 H) 1.26-1.38 (m, 1
H) 1.66 (d, J = 8.75 Hz, 2 H) 1.80 (br. s., 1 H) 1.89-1.98 (m, 2 H)
2.78-2.89 (m, 1 H) 3.12 (d, J = 11.74 Hz, 1 H) 3.68 (br. s., 1 H)
3.83 (br. s., 1 H) 3.88-3.97 (m, 1 H) 4.35-4.42 (m, 1 H) 4.75 (t, J
= 6.26 Hz, 2 H) 5.15 (t, J = 7.29 Hz, 2 H) 7.31 (d, J = 9.54 Hz, 2
H) 7.56 (d, J = 4.99 Hz, 1 H) 8.02 (t, J = 8.80 Hz, 1 H) 8.40 (d, J
= 5.33 Hz, 1 H) 8.50 (d, J = 5.04 Hz, 1 H) 8.82 (s, 1 H) 164
.sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.46 (d, J = 6.70
Hz, 3 H) 2.39 (d, J = 5.62 Hz, 1 H) 2.85 (t, J = 12.84 Hz, 1 H)
3.59 (br. s., 1 H) 3.80-3.90 (m, 1 H) 3.98-4.14 (m, 4 H) 4.25 (d, J
= 12.03 Hz, 1 H) 5.08-5.17 (m, 1 H) 5.32 (d, J = 11.88 Hz, 1 H)
5.46 (br. s., 1 H) 6.88 (d, J = 9.05 Hz, 1 H) 7.55 (d, J = 6.70 Hz,
1 H) 7.69 (d, J = 10.12 Hz, 1 H) 8.10 (t, J = 8.61 Hz, 1 H) 8.33
(d, J = 6.46 Hz, 1 H) 8.39 (dd, J = 8.58, 3.45 Hz, 1 H) 8.97 (s, 1
H) 9.19 (s, 1 H) 9.55 (s, 1 H) 165 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 1.12 (d, J = 6.65 Hz, 3 H) 1.54-1.70 (m,
2 H) 1.72-1.84 (m, 1 H) 1.87-2.00 (m, 2 H) 2.71-2.90 (m, 2 H)
3.01-3.17 (m, 1 H) 3.64-3.71 (m, 1 H) 3.76-3.97 (m, 5 H) 6.74-6.83
(m, 2 H) 7.54-7.63 (m, 1 H) 7.92-8.02 (m, 1 H) 8.31-8.39 (m, 1 H)
8.46-8.52 (m, 1 H) 8.89-8.96 (m, 1 H) 166 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 1.02-1.18 (m, 3 H) 1.50-1.75 (m, 3 H)
1.87-2.03 (m, 2 H) 2.70-2.90 (m, 2 H) 3.02-3.18 (m, 1 H) 3.34-3.50
(m, 1 H) 3.66 (br. s., 1 H) 3.76-3.96 (m, 2 H) 7.08-7.18 (m, 3 H)
7.47-7.68 (m, 2 H) 8.46 (d, J = 5.48 Hz, 1 H) 9.20 (s, 1 H) 167
.sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.94 (d, J = 6.70
Hz, 3 H) 1.24-1.36 (m, 1 H) 1.41-1.58 (m, 1 H) 1.62-1.99 (m, 5 H)
2.11 (br. s., 1 H) 2.37-2.62 (m, 4 H) 3.03-3.17 (m, 1 H) 3.73 (s, 3
H) 4.09 (br. s., 1 H) 7.17 (d, J = 9.10 Hz, 1 H) 7.36 (d, J = 9.54
Hz, 2 H) 7.68 (d, J = 5.33 Hz, 1 H) 8.02 (t, J = 8.68 Hz, 1 H) 8.40
(dd, J = 8.63, 3.74 Hz, 1 H) 8.49 (d, J = 5.28 Hz, 1 H) 8.91 (s, 1
H) 168 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 8.85 (s, 1
H), 8.58 (d, J = 5.48 Hz, 1 H), 8.38 (dd, J = 8.61, 3.52 Hz, 1 H),
8.07 (s, 1 H), 7.99 (t, J = 8.80 Hz, 1 H), 7.90 (s, 1 H), 7.82 (d,
J = 5.09 Hz, 1 H), 6.84 (d, J = 10.17 Hz, 2 H), 4.98 (br. s., 1 H),
4.22 (br. s., 2 H), 3.89 (d, J = 12.52 Hz, 1 H), 3.78 (br. s., 2
H), 3.44 (s, 3 H), 2.64-2.74 (m, 2 H), 2.29 (br. s., 1 H),
2.02-2.18 (m, 2 H), 1.81 (d, J = 13.69 Hz, 1 H), 0.67 (d, J = 6.65
Hz, 3 H) 169 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 8.84
(s, 1 H), 8.65 (s, 1 H), 8.60 (d, J = 5.09 Hz, 1 H), 8.08 (s, 1 H),
7.90 (s, 1 H), 7.85 (d, J = 5.48 Hz, 1 H), 7.63 (t, J = 7.83 Hz, 1
H), 7.25 (t, J = 8.80 Hz, 2 H), 5.00 (br. s., 1 H), 3.95 (d, J =
12.52 Hz, 1 H), 2.63-2.76 (m, 1 H), 2.38 (br. s., 1 H), 2.06-2.21
(m, 2 H), 1.84 (d, J = 12.91 Hz, 1 H), 0.70 (d, J = 6.65 Hz, 3 H)
170 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.94 (d, J =
6.60 Hz, 3 H) 1.50 (q, J = 13.17 Hz, 1 H) 1.65-1.84 (m, 4 H)
1.85-2.01 (m, 2 H) 2.09 (d, J = 11.59 Hz, 2 H) 3.04-3.18 (m, 2 H)
3.35-3.51 (m, 2 H) 3.64 (t, J = 9.24 Hz, 2 H) 3.73 (s, 3 H)
3.93-4.02 (m, 2 H) 4.09 (br. s., 1 H) 4.71 (br. s., 1 H) 6.86 (d, J
= 10.37 Hz, 2 H) 7.18 (d, J = 10.07 Hz, 1 H) 7.70 (d, J = 5.43 Hz,
1 H) 7.99 (t, J = 8.61 Hz, 1 H) 8.36 (dd, J = 8.73, 3.94 Hz, 1 H)
8.50 (d, J = 5.33 Hz, 1 H) 8.93 (s, 1 H) 171 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 0.67 (d, J = 6.70 Hz, 3 H) 1.68-1.87 (m,
3 H) 2.00-2.20 (m, 4 H) 2.30 (br. s., 1 H) 2.63-2.77 (m, 1 H) 3.63
(t, J = 8.95 Hz, 2 H) 3.84-4.03 (m, 3 H) 4.66-4.76 (m, 1 H) 4.98
(br. s., 1 H) 6.86 (d, J = 10.17 Hz, 2 H) 7.82 (d, J = 5.18 Hz, 1
H) 7.89 (s, 1 H) 7.99 (t, J = 8.66 Hz, 1 H) 8.07 (s, 1 H) 8.37 (dd,
J = 8.66, 3.86 Hz, 1 H) 8.58 (d, J = 5.28 Hz, 1 H) 8.80 (s, 1 H)
172 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.65 (d, J =
6.60 Hz, 3 H) 1.73-1.92 (m, 2 H) 2.00-2.17 (m, 3 H) 2.27 (br. s., 1
H) 2.37-2.60 (m, 4 H) 2.63-2.76 (m, 1 H) 3.86 (br. s., 1 H) 4.96
(br. s., 2 H) 7.35 (d, J = 9.63 Hz, 2 H) 7.78 (d, J = 5.28 Hz, 1 H)
7.89 (s, 1 H) 7.99-8.08 (m, 2 H) 8.37-8.43 (m, 1 H) 8.56 (d, J =
5.43 Hz, 1 H) 8.79 (s, 1 H) 174 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 0.50 (d, J = 7.04 Hz, 3 H) 1.84-1.96 (m,
1 H) 2.54-2.66 (m, 1 H) 3.04-3.15 (m, 1 H) 3.25-3.45 (m, 3 H) 3.61
(s, 3 H) 3.76-3.83 (m, 1 H) 7.01-7.16 (m, 3 H) 7.32-7.41 (m, 1 H)
7.46-7.58 (m, 1 H) 8.22-8.32 (m, 1 H) 9.33-9.42 (m, 1 H) 175
.sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.87 (d, J = 6.65
Hz, 3 H) 1.39-1.53 (m, 1 H) 1.62-1.99 (m, 4 H) 3.04-3.16 (m, 1 H)
3.32-3.37 (m, 1 H) 3.70 (s, 3 H) 4.03-4.11 (m, 1 H) 7.08-7.21 (m, 4
H) 7.49-7.60 (m, 1 H) 7.80 (d, J = 5.48 Hz, 1 H) 8.48 (m, 1 H) 9.29
(s, 1 H) 176 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.52
(d, J = 6.65 Hz, 3 H) 1.65-1.75 (m, 1 H) 1.90-2.16 (m, 3 H)
2.59-2.73 (m, 1 H) 3.26-3.35 (m, 1 H) 3.71-3.81 (m, 1 H) 4.91-4.97
(m, 1 H) 7.02-7.13 (m, 3 H) 7.40-7.51 (m, 1 H) 7.79 (s, 2 H) 8.00
(s, 1 H) 8.45-8.52 (m, 1 H) 9.12 (s, 1 H) 177 .sup.1H NMR, (400
MHz, <cd3od>) .delta. ppm 9.46 (s, 1 H), 8.48 (dd, J = 8.6,
3.9 Hz, 1 H), 8.42 (d, J = 6.3 Hz, 1 H), 8.08 (t, J = 8.8 Hz, 1 H),
7.39-7.53 (m, 3 H), 3.86-4.09 (m, 5 H), 3.72 (s, 3 H), 3.57 (dd, J
= 8.4, 3.7 Hz, 2 H), 3.37-3.47 (m, 1 H), 3.19-3.28 (m, 1 H), 2.77
(t, J = 12.9 Hz, 1 H), 2.10-2.36 (m, 3 H), 1.64-1.86 (m, 2 H), 0.56
(d, J = 6.7 Hz, 3 H) 178 .sup.1H NMR, (400 MHz, <cd3od>)
.delta. ppm 8.99 (s, 1 H), 8.54 (d, J = 5.5 Hz, 1 H), 8.42 (dd, J =
9.0, 3.9 Hz, 1 H), 8.04 (t, J = 8.8 Hz, 1 H), 7.75 (d, J = 5.5 Hz,
1 H), 7.39 (d, J = 9.8 Hz, 2 H), 7.19 (d, J = 9.8 Hz, 1 H), 4.12
(d, J = 9.0 Hz, 1 H), 3.84-4.04 (m, 4 H), 3.74 (s, 3 H), 3.39 (d, J
= 3.5 Hz, 1 H), 3.14 (t, J = 12.1 Hz, 1 H), 2.12-2.28 (m, 2 H),
1.86-2.03 (m, 2 H), 1.64-1.85 (m, 4 H), 1.51 (q, J = 12.8 Hz, 1 H),
0.94 (d, J = 6.7 Hz, 3 H) 179 .sup.1H NMR, (400 MHz, <cd3od>)
.delta. ppm 8.81 (s, 1 H), 8.60 (d, J = 5.5 Hz, 1 H), 8.42 (dd, J =
8.6, 3.9 Hz, 1 H), 8.08 (s, 1 H), 8.04 (t, J = 8.6 Hz, 1 H), 7.91
(s, 1 H), 7.84 (d, J = 5.5 Hz, 1 H), 7.38 (d, J = 9.8 Hz, 2 H),
5.00 (t, J = 3.9 Hz, 1 H), 3.81-4.04 (m, 4 H), 3.37 (br. s., 1 H),
2.64-2.80 (m, 1 H), 2.26-2.39 (m, 1 H), 2.02-2.25 (m, 4 H), 1.82
(d, J = 13.3 Hz, 1 H), 1.68 (d, J = 13.3 Hz, 2 H), 0.67 (d, J = 6.7
Hz, 3 H) 180 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 9.03
(s, 1 H), 8.55 (d, J = 5.5 Hz, 1 H), 8.41 (dd, J = 8.6, 3.9 Hz, 1
H), 8.02 (t, J = 8.6 Hz, 1 H), 7.80 (d, J = 5.9 Hz, 1 H), 7.20 (d,
J = 9.8 Hz, 1 H), 7.07 (d, J = 9.4 Hz, 2 H), 4.12 (d, J = 9.4 Hz, 1
H), 3.74 (s, 3 H), 3.36-3.47 (m, 1 H), 3.09-3.23 (m, 1 H), 2.49 (s,
3 H), 1.65-2.04 (m, 4 H), 1.52 (q, J = 13.2 Hz, 1 H), 0.95 (d, J =
7.0 Hz, 3 H) 181 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm
0.68 (d, J = 6.70 Hz, 3 H) 1.76-1.88 (m, 1 H) 2.02-2.20 (m, 2 H)
2.23-2.38 (m, 1 H) 2.49 (s, 3 H) 2.64-2.80 (m, 1
H) 3.35-3.39 (m, 1 H) 3.90 (dt, J = 12.72, 4.43 Hz, 1 H) 4.97-5.03
(m, 1 H) 7.07 (d, J = 9.44 Hz, 2 H) 7.84 (d, J = 5.48 Hz, 1 H) 7.91
(d, J = 0.98 Hz, 1 H) 8.02 (t, J = 8.71 Hz, 1 H) 8.09 (d, J = 1.03
Hz, 1 H) 8.41 (dd, J = 8.61, 3.96 Hz, 1 H) 8.60 (d, J = 5.48 Hz, 1
H) 8.84 (s, 1 H) 182 .sup.1H NMR, (400 MHz, <cd3od>) .delta.
ppm 0.94 (d, J = 6.75 Hz, 3 H) 1.51 (q, J = 12.78 Hz, 1 H) 1.71 (d,
J = 13.64 Hz, 1 H) 1.80 (q, J = 12.41 Hz, 1 H) 1.88-2.02 (m, 2 H)
3.10-3.20 (m, 1 H) 3.41 (dt, J = 12.47, 3.55 Hz, 1 H) 3.74 (s, 3 H)
4.08-4.14 (m, 1 H) 4.36-4.45 (m, 1 H) 4.77 (td, J = 6.28, 1.47 Hz,
2 H) 5.16 (dd, J = 8.22, 6.21 Hz, 2 H) 7.20 (d, J = 9.73 Hz, 1 H)
7.33 (d, J = 9.10 Hz, 2 H) 7.78 (d, J = 5.58 Hz, 1 H) 8.04 (t, J =
8.71 Hz, 1 H) 8.42 (dd, J = 8.68, 3.94 Hz, 1 H) 8.55 (d, J = 5.53
Hz, 1 H) 8.98 (s, 1 H) 183 .sup.1H NMR, (400 MHz, <cd3od>)
.delta. ppm 1.13 (d, J = 6.85 Hz, 3 H) 1.53-1.85 (m, 5 H) 1.85-2.00
(m, 2 H) 2.00-2.15 (m, 2 H) 2.70-2.94 (m, 2 H) 3.03-3.16 (m, 2 H)
3.56-3.70 (m, 3 H) 3.75-3.86 (m, 1 H) 3.87-4.05 (m, 3 H) 4.70 (dt,
J = 8.03, 4.22 Hz, 1 H) 6.84 (d, J = 10.12 Hz, 2 H) 7.57 (d, J =
5.48 Hz, 1 H) 7.97 (t, J = 8.71 Hz, 1 H) 8.35 (dd, J = 8.63, 3.94
Hz, 1 H) 8.49 (d, J = 5.48 Hz, 1 H) 8.86 (s, 1 H) 184 .sup.1H NMR,
(400 MHz, <cd3od>) .delta. ppm 1.13 (d, J = 6.85 Hz, 3 H)
1.58-1.73 (m, 2 H) 1.73-2.02 (m, 4 H) 2.05-2.23 (m, 1 H) 2.33-2.63
(m, 4 H) 2.72-2.92 (m, 2 H) 3.02-3.18 (m, 1 H) 3.69 (s, 1 H) 3.82
(dt, J = 9.06, 5.42 Hz, 1 H) 3.94 (td, J = 8.63, 5.09 Hz, 1 H) 7.36
(d, J = 9.49 Hz, 2 H) 7.59 (d, J = 5.48 Hz, 1 H) 8.03 (t, J = 8.71
Hz, 1 H) 8.40 (dd, J = 8.63, 3.94 Hz, 1 H) 8.51 (d, J = 5.43 Hz, 1
H) 8.92 (s, 1 H) 185 .sup.1H NMR, (400 MHz, <cd3od>) .delta.
ppm 0.69 (d, J = 6.75 Hz, 3 H) 1.84 (d, J = 13.69 Hz, 1 H)
1.90-2.10 (m, 1 H) 2.16 (d, J = 12.47 Hz, 1 H) 2.32 (br. s., 1 H)
2.50-2.72 (m, 1 H) 3.94 (dt, J = 12.89, 4.56 Hz, 1 H) 5.04 (t, J =
4.16 Hz, 1 H) 7.22 (t, J = 8.29 Hz, 2 H) 7.58-7.70 (m, 1 H) 7.75
(d, J = 5.43 Hz, 1 H) 8.04 (t, J = 8.71 Hz, 1 H) 8.42 (dd, J =
8.66, 3.96 Hz, 1 H) 8.57 (d, J = 5.38 Hz, 1 H) 8.75 (s, 1 H) 8.81
(s, 1 H) 186 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.67
(d, J = 6.70 Hz, 3 H) 0.76-0.91 (m, 2 H) 0.98-1.08 (m, 2 H)
1.72-1.84 (m, 1 H) 1.96-2.17 (m, 3 H) 2.17-2.36 (m, 1 H) 2.58-2.78
(m, 1 H) 3.84 (dt, J = 12.67, 4.45 Hz, 1 H) 7.22 (t, J = 8.27 Hz, 2
H) 7.64 (tt, J = 8.49, 6.47 Hz, 1 H) 7.77 (s, 1 H) 7.82 (d, J =
5.58 Hz, 1 H) 8.04 (t, J = 8.73 Hz, 1 H) 8.42 (dd, J = 8.66, 3.96
Hz, 1 H) 8.59 (d, J = 5.38 Hz, 1 H) 8.86 (s, 1 H) 187 .sup.1H NMR,
(400 MHz, <cd3od>) .delta. ppm 0.69 (d, J = 6.65 Hz, 3 H)
1.80 (d, J = 13.30 Hz, 1 H) 2.00-2.14 (m, 2 H) 2.17 (s, 3 H)
2.20-2.36 (m, 1 H) 2.60-2.80 (m, 1 H) 3.87 (dt, J = 12.63, 4.32 Hz,
1 H) 4.91 (d, J = 6.36 Hz, 1 H) 5.19 (s, 1 H) 5.75 (s, 1 H) 7.22
(t, J = 8.29 Hz, 2 H) 7.63 (tt, J = 8.48, 6.49 Hz, 1 H) 7.84 (br.
s., 1 H) 8.03 (t, J = 8.71 Hz, 1 H) 8.09 (d, J = 1.86 Hz, 1 H) 8.41
(dd, J = 8.66, 3.91 Hz, 1 H) 8.54-8.63 (m, 1 H) 8.76-8.91 (m, 1 H)
188 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.66 (d, J =
6.75 Hz, 3 H) 1.63 (s, 6 H) 1.71-1.82 (m, 1 H) 1.90-2.18 (m, 2 H)
2.24 (d, J = 12.57 Hz, 1 H) 2.61-2.79 (m, 1 H) 3.85 (dt, J = 12.64,
4.34 Hz, 1 H) 7.21 (t, J = 8.29 Hz, 2 H) 7.57-7.70 (m, 1 H) 7.81
(d, J = 5.48 Hz, 1 H) 7.87 (s, 1 H) 8.02 (t, J = 8.71 Hz, 1 H) 8.41
(dd, J = 8.63, 3.99 Hz, 1 H) 8.57 (d, J = 5.58 Hz, 1 H) 8.82 (s, 1
H) 189 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.97 (s, 3
H) 1.45-1.60 (m, 2 H) 1.60-1.71 (m, 1 H) 1.80-1.90 (m, 1 H)
1.91-2.03 (m, 1 H) 2.97-3.11 (m, 1 H) 3.26-3.31 (m, 1 H) 3.60-3.67
(m, 1 H) 4.08-4.17 (m, 1 H) 4.35-4.45 (m, 1 H) 7.06-7.17 (m, 2 H)
7.48-7.65 (m, 2 H) 7.90-7.98 (m, 1 H) 8.28-8.36 (m, 1 H) 8.41-8.47
(m, 1 H) 8.91-8.97 (m, 1 H) 190 .sup.1H NMR, (400 MHz,
<cd3od>) .delta. ppm 1.02-1.10 (m, 3 H) 1.52-1.67 (m, 2 H)
1.68-1.80 (m, 1 H) 1.88-1.96 (m, 1 H) 1.97-2.10 (m, 1 H) 3.04-3.16
(m, 1 H) 3.35-3.41 (m, 1 H) 3.67-3.73 (m, 1 H) 4.16-4.25 (m, 1 H)
4.43-4.52 (m, 1 H) 7.14-7.24 (m, 2 H) 7.56-7.69 (m, 2 H) 7.97-8.05
(m, 1 H) 8.36-8.43 (m, 1 H) 8.47-8.55 (m, 1 H) 8.94-9.00 (m, 1 H)
191 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.88-0.97 (m,
3 H) 1.42-1.55 (m, 1 H) 1.63-1.80 (m, 2 H) 1.85-1.98 (m, 2 H)
3.03-3.16 (m, 1 H) 3.36-3.41 (m, 1 H) 3.66-3.74 (m, 3 H) 3.85-3.92
(m, 3 H) 4.04-4.12 (m, 1 H) 6.78 (s, 1 H) 7.64-7.72 (m, 1 H)
7.92-8.01 (m, 1 H) 8.31-8.39 (m, 1 H) 8.45-8.51 (m, 1 H) 8.86-8.94
(m, 1 H) 192 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm 0.64
(d, J = 6.65 Hz, 3 H) 1.73-1.85 (m, 1 H) 2.00-2.16 (m, 2 H)
2.21-2.34 (m, 1 H) 2.61-2.76 (m, 1 H) 3.34-3.41 (m, 1 H) 3.88 (s, 4
H) 4.95-5.02 (m, 1 H) 6.74-6.84 (m, 2 H) 7.79-7.85 (m, 1 H)
7.85-7.91 (m, 1 H) 7.93-8.01 (m, 1 H) 8.02-8.09 (m, 1 H) 8.31-8.40
(m, 1 H) 8.53-8.61 (m, 1 H) 8.81-8.89 (m, 1 H) 193 .sup.1H NMR,
(400 MHz, <cd3od>) .delta. ppm 0.43 (d, J = 6.65 Hz, 3 H)
2.44-2.58 (m, 1 H) 3.57-3.67 (m, 1 H) 3.76-3.84 (m, 1 H) 3.93 (s, 3
H) 3.97-4.13 (m, 2 H) 4.19-4.29 (m, 1 H) 5.06-5.12 (m, 1 H)
6.79-6.89 (m, 2 H) 7.52-7.60 (m, 1 H) 7.84-7.91 (m, 1 H) 7.95-8.04
(m, 1 H) 8.07-8.14 (m, 1 H) 8.34-8.44 (m, 2 H) 9.07-9.13 (m, 1 H)
194 .sup.1H NMR, (400 MHz, <cd3od>) .delta. ppm -0.02-0.04
(m, 3 H) 1.94-2.07 (m, 1 H) 3.01-3.11 (m, 2 H) 3.16-3.24 (m, 1 H)
3.35-3.44 (m, 1 H) 3.50-3.59 (m, 1 H) 3.72-3.82 (m, 1 H) 6.78 (s, 1
H) 7.12-7.29 (m, 2 H) 7.49-7.52 (m, 1 H) 7.70-7.74 (m, 1 H)
7.97-8.04 (m, 1 H) 8.92-8.97 (m, 1 H) 198 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm 0.59 (d, J = 6.65 Hz, 3 H) 1.16
(dd, J = 8.41, 6.85 Hz, 6 H) 2.06 (d, J = 3.91 Hz, 1 H) 3.07-3.33
(m, 5 H) 3.56 (s, 3 H) 3.86 (d, J = 9.78 Hz, 1 H) 4.18 (dq, J =
13.60, 6.68 Hz, 1 H) 7.19 (br. s., 1 H) 7.28-7.52 (m, 4 H) 7.78
(dd, J = 8.61, 1.96 Hz, 1 H) 7.86-8.06 (m, 4 H) 8.28-8.52 (m, 3 H)
9.16 (s, 1 H) 10.24 (s, 1 H) 199 .sup.1H NMR (400 MHz,
<dmso-.sub.d6>) .delta. ppm 0.45 (d, J = 7.04 Hz, 3 H)
1.74-2.02 (m, 3 H) 2.07-2.41 (m, 2 H) 2.84-2.99 (m, 1 H) 3.08-3.22
(m, 1 H) 3.30 (d, J = 14.48 Hz, 2 H) 3.48 (d, J = 9.39 Hz, 1 H)
3.56 (s, 3 H) 3.69 (t, J = 11.15 Hz, 2 H) 3.79-4.00 (m, 3 H) 7.28
(d, J = 6.26 Hz, 1 H) 7.36 (d, J = 10.17 Hz, 1 H) 7.47 (d, J = 9.39
Hz, 2 H) 8.03 (br. s., 3 H) 8.23 (t, J = 8.80 Hz, 1 H) 8.29-8.46
(m, 2 H) 9.00 (br. s., 1 H) 10.36 (s, 1 H) 200 .sup.1H NMR (400
MHz, <dmso-.sub.d6>) .delta. ppm 0.75 (d, J = 6.65 Hz, 3 H)
1.06-1.21 (m, 6 H) 1.42 (d, J = 12.91 Hz, 1 H) 1.51-1.71 (m, 2 H)
1.77 (d, J = 12.13 Hz, 2 H) 2.99 (br. s., 1 H) 3.32 (br. s., 1 H)
3.52-3.68 (m, 3 H) 3.85-3.99 (m, 1 H) 4.09 (dq, J = 13.79, 6.75 Hz,
1 H) 7.31-7.44 (m, 3 H) 7.69-7.85 (m, 5 H) 7.88 (ddd, J = 8.31,
4.60, 2.35 Hz, 1 H) 8.32 (d, J = 7.43 Hz, 1 H) 8.40 (dd, J = 7.83,
1.96 Hz, 1 H) 8.49 (d, J = 5.09 Hz, 1 H) 8.88 (s, 1 H) 10.36 (s, 1
H) 201 .sup.1H NMR (400 MHz, <dmso-.sub.d6>) .delta. ppm 0.76
(d, J = 6.65 Hz, 3 H) 1.29-1.44 (m, 1 H) 1.44-1.80 (m, 4 H)
1.80-1.95 (m, 2 H) 2.18 (td, J = 13.21, 5.28 Hz, 1 H) 2.28 (td, J =
13.01, 5.67 Hz, 1 H) 2.79-2.93 (m, 1 H) 3.20 (br. s., 1 H)
3.53-3.63 (m, 3 H) 3.68 (t, J = 11.35 Hz, 2 H) 3.79-3.98 (m, 4 H)
7.33-7.53 (m, 3 H) 7.67-7.90 (m, 4 H) 8.11-8.23 (m, 1 H) 8.25-8.35
(m, 1 H) 8.48 (d, J = 5.48 Hz, 1 H) 8.54-8.63 (m, 1 H) 10.37-10.47
(m, 1 H) 202 .sup.1H NMR (400 MHz, <cdcl.sub.3>) .delta. ppm
0.87 (br. s., 3 H) 1.27 (t, J = 6.65 Hz, 3 H) 1.45-1.68 (m, 2 H)
1.80 (br. s., 1 H) 2.05 (br. s., 1 H) 2.24 (br. s., 1 H) 2.99 (br.
s., 1 H) 3.41-3.68 (m, 6 H) 4.23 (br. s., 1 H) 4.51-4.66 (m, 2 H)
6.79-7.19 (m, 3 H) 7.70-8.11 (m, 2 H) 8.20-8.64 (m, 3 H) 9.40-9.64
(m, 1 H) 10.17-10.40 (m, 1 H) 203 .sup.1H NMR (400 MHz,
<cdcl.sub.3>) .delta. ppm 0.42-0.71 (m, 3 H) 1.28 (br. s., 3
H) 2.06 (br. s., 1 H) 2.88 (br. s., 1 H) 3.25 (br. s., 1 H)
3.47-3.83 (m, 8 H) 4.10 (br. s., 1 H) 4.62 (br. s., 2 H) 6.83-7.06
(m, 1 H) 7.18 (br. s., 2 H) 7.48 (br. s., 1 H) 7.75 (br. s., 1 H)
8.34 (br. s., 2 H) 9.45-9.68 (m, 1 H) 10.05 (br. s., 1 H)
Pim1, Pim2, Pim3 AlphaScreen Assays
[0566] Pim 1, Pim 2 & Pim 3 AlphaScreen assays using high ATP
(11-125.times.ATP Km) were used to determine the biochemical
activity of the inhibitors. The activity of Pim 1, Pim 2, & Pim
3 is measured using a homogeneous bead based system quantifying the
amount of phosphorylated peptide substrate resulting from
kinase-catalyzed phosphoryl transfer to a peptide substrate.
Compounds to be tested are dissolved in 100% DMSO and directly
distributed to a white 384-well plate at 0.25 .mu.l per well. To
start the reaction, 5 .mu.l of 100 nM Bad peptide
(Biotin-AGAGRSRHSSYPAGT-OH (SEQ ID NO:1)) and ATP (concentrations
described below) in assay buffer (50 mM Hepes, pH=7.5, 5 mM
MgCl.sub.2, 0.05% BSA, 0.01% Tween-20, 1 mM DTT) is added to each
well. This is followed by the addition of 5 .mu.l/well of Pim 1,
Pim 2 or Pim 3 kinase in assay buffer (concentrations described
below). Final assay concentrations (described below) are in 2.5%
DMSO. The reactions are performed for .about.2 hours, then stopped
by the addition of 10 .mu.l of 0.75 .mu.g/ml anti-phospho Ser/Thr
antibody (Cell Signaling), 10 .mu.g/ml Protein A AlphaScreen beads
(Perkin Elmer), and 10 .mu.g/ml streptavidin coated AlphaScreen
beads in stop/detection buffer (50 mM EDTA, 95 mM Tris, pH=7.5,
0.01% Tween-20). The stopped reactions are incubated overnight in
the dark. The phosphorylated peptide is detected via an oxygen
anion initiated chemiluminescence/fluorescence cascade using the
Envision plate reader (Perkin Elmer).
TABLE-US-00005 AlphaScreen Assay Conditions b-BAD ATP Km Enzyme
Enzyme conc. peptide conc. ATP conc. (app) source (nM) (nM) (uM)
(uM) Pim 1 (INV) 0.0025 50 2800 246 Pim 2 (INV) 0.01 50 500 4 Pim 3
(NVS) 0.005 50 2500 50
[0567] Compounds of the foregoing examples were tested by the Pim
1, Pim 2 & Pim 3 AlphaScreen assays and found to exhibit an
IC.sub.50 values as shown in Table 4 below. IC.sub.50, the half
maximal inhibitory concentration, represents the concentration of
test compound that is required for 50% inhibition of its target in
vitro.
Cell Proliferation Assay
[0568] KMS 11 (human myeloma cell line), were cultured in IMDM
supplemented with 10% FBS, sodium pyruvate and antibiotics. Cells
were plated in the same medium at a density of 2000 cells per well
into 96 well tissue culture plates, with outside wells vacant, on
the day of assay.
[0569] Test compounds supplied in DMSO were diluted into DMSO at
500 times the desired final concentrations before dilution into
culture media to 2 times final concentrations. Equal volumes of
2.times. compounds were added to the cells in 96 well plates and
incubated at 37.degree. C. for 3 days.
[0570] After 3 days plates were equilibrated to room temperature
and equal volume of CellTiter-Glow Reagent (Promega) was added to
the culture wells. The plates were agitated briefly and luminescent
signal was measured with luminometer. The percent inhibition of the
signal seen in cells treated with DMSO alone vs. cells treated with
control compound was calculated and used to determine EC.sub.50
values (i.e., the concentration of a test compound that is required
to obtain 50% of the maximum effect in the cells) for tested
compounds, as shown in Table 4.
[0571] Using the procedures of the Pim1, Pim2, Pim3 AlphaScreen
Assays the IC.sub.50 concentrations of compounds of the previous
examples were determined as shown in the Table 4.
[0572] Using the procedures of Cell Proliferation Assay, the
EC.sub.50 concentrations of compounds of the examples were
determined in KMS 11 cells as shown in Table 4.
TABLE-US-00006 TABLE 4 Pim1 Pim2 Pim3 KMS11 Ex# IC50 .mu.M IC50
.mu.M IC50 .mu.M EC50.mu.M 1 0.00006 0.00253 0.00252 0.094 2
0.00036 0.03007 0.01572 1.008 3 0.00014 0.04942 0.03053 1.130 4
0.00006 0.00428 0.00154 0.148 5 0.00003 0.00124 0.00064 0.033 6
0.00024 0.00681 0.01100 3.090 7 0.00004 0.00139 0.00086 0.040 8
0.00035 0.02050 0.01423 0.499 9 0.00040 0.05937 0.01778 0.670 10
0.00057 0.03407 0.02088 1.833 11 0.00038 0.03331 0.01956 4.179 12
0.00021 0.01607 0.01572 0.716 13 0.00009 0.00482 0.00509 0.507 14
0.00004 0.00172 0.00102 0.045 15 0.00007 0.00356 0.00207 0.146 16
0.17209 21.7 8.3 >10 17 0.00256 0.38852 0.15521 8.260 18 0.00005
0.00086 0.00099 0.267 19 0.00017 0.00259 0.00321 0.287 20 0.00014
0.00668 0.00678 0.476 21 0.00005 0.00766 0.00092 0.756 22 0.00037
0.01388 0.01165 0.530 23 0.02746 1.4 1.0 >10 24 0.06819 2.7 1.7
>10 25 0.00063 0.02504 0.01580 2.867 26 0.14276 7.1 4.8 >10
27 0.00212 0.04198 0.05333 1.751 28 0.00680 0.23750 0.15437 4.874
29 0.00020 0.00944 0.00888 0.768 30 0.00072 0.07280 0.04426 9.686
31 0.00021 0.01045 0.01240 0.633 32 0.00297 0.10611 0.06386 7.958
33 0.00002 0.00068 0.00061 0.072 34 0.00076 0.04724 0.03991 3.753
35 0.00014 0.00794 0.00524 0.365 36 0.00059 0.02943 0.02876 0.979
37 0.00009 0.00300 0.00503 0.178 38 0.00056 0.04551 0.04371 5.949
39 0.00039 0.03092 0.02182 2.989 40 0.00017 0.01241 0.00875 0.964
41 0.02032 5.980 42 0.00004 0.00196 0.00100 0.063 43 0.00586
0.32741 0.11823 4.577 44 0.00338 0.28911 0.22295 >10 45 0.64280
>25 >25 >10 46 0.00020 0.02139 0.00304 0.348 47 0.17919
15.8 2.7 >10 48 0.01063 0.42676 0.41111 9.416 49 0.00003 0.00120
0.00116 0.031 50 0.00024 0.01707 0.00535 0.470 51 0.00177 0.06911
0.03579 0.970 52 0.00850 0.72623 0.59094 4.386 53 0.00013 0.01432
0.01054 0.363 54 0.00009 0.00468 0.00174 0.891 55 0.00013 0.01118
0.00629 0.553 56 0.00002 0.00151 0.00062 0.036 57 0.00017 0.00647
0.00571 0.462 58 0.00002 0.00151 0.00049 0.168 59 0.01678 0.86628
0.58283 7.309 60 0.00022 0.01356 0.01080 0.202 61 0.00003 0.00107
0.00049 0.041 62 0.00003 0.00290 0.00128 0.106 63 0.00630 0.36441
0.09475 4.486 64 0.00001 0.00100 0.00022 0.044 65 0.04056 2.8 1.1
>10 66 0.00003 0.00118 0.00073 0.224 67 0.00148 0.07360 0.07771
3.205 68 0.00028 0.01196 0.01485 0.847 69 0.00004 0.00117 0.00040
0.114 70 0.00075 0.01070 0.00819 0.144 71 0.00004 0.00170 0.00054
0.156 72 0.00003 0.00083 0.00043 0.065 73 0.00002 0.00137 0.00091
0.069 74 0.04346 2.48310 1.50872 7.843 75 0.00008 0.00337 0.00308
0.147 76 0.13046 6.6 4.3 >10 77 0.00004 0.00144 0.00162 0.080 78
0.02538 1.1 1.0 >10 79 0.00028 0.00257 0.00536 0.132 80 0.00003
0.00082 0.00079 0.050 81 0.00004 0.00089 0.00074 0.036 82 0.00003
0.00062 0.00032 0.048 83 0.00001 0.00063 0.00064 0.037 84 0.00133
0.06413 0.04181 3.201 85 0.00003 0.00113 0.00096 0.057 86 0.00308
0.09956 0.06764 5.155 87 0.00003 0.00074 0.00068 0.166 88 0.00003
0.00237 0.00124 0.093 89 0.01317 1.47124 0.58270 3.990 90 0.03105
1.9 1.0 >10 91 0.00005 0.00226 0.00264 0.128 92 0.00010 0.00471
0.00369 0.175 93 0.00004 0.00368 0.00188 0.154 94 0.00003 0.00235
0.00279 0.356 95 0.00002 0.00138 0.00143 0.289 96 0.00004 0.00242
0.00507 0.186 97 0.00004 0.00133 0.00220 0.051 98 0.00004 0.00172
0.00158 0.074 99 0.00001 0.00108 0.00125 0.051 100 0.00006 0.00330
0.00373 0.165 101 0.00002 0.00301 0.00145 0.212 102 0.00003 0.00218
0.00210 0.454 103 0.00212 0.13033 0.10610 3.451 104 0.00535 0.13520
0.19792 >10 105 0.00006 0.00374 0.00161 0.117 106 0.00031
0.02440 0.01673 0.792 107 0.00005 0.00769 0.00542 0.364 108 0.00008
0.00234 0.00581 0.155 109 0.00040 0.00161 0.01117 0.616 110 0.00009
0.01038 0.01791 0.825 111 0.00008 0.00074 0.00141 0.169 112 0.00008
0.00737 0.00415 0.258 113 0.00250 0.12003 0.11490 1.826 114 0.00188
0.053 115 0.00007 0.00248 0.00144 0.315 116 0.00012 0.00578 0.01210
0.323 117 0.00003 0.00153 0.00310 0.026 118 0.00002 0.00124 0.00174
2.455 119 0.00007 0.00907 0.00525 0.316 120 0.00140 0.14659 0.06752
6.060 121 0.00005 0.00593 0.01043 0.625 122 0.00005 0.00385 0.00384
0.321 123 0.00003 0.00153 0.00153 0.054 124 0.00005 0.00403 0.00202
0.040 125 0.00004 0.00179 0.00239 0.046 126 0.00070 0.00256 0.00778
0.267 127 0.00005 0.00198 0.00451 0.076 128 0.00024 0.00121 0.00411
2.099 129 0.00005 0.00246 0.00544 0.107 130 0.00003 0.00254 0.00170
0.109 131 0.00005 0.00255 0.00149 0.135 132 0.00004 0.00145 0.00108
0.016 133 0.00057 0.02866 0.01050 2.304 134 0.00054 0.02207 0.01019
0.468 135 0.00002 0.00078 0.00026 0.193 136 0.00004 0.00147 0.00249
0.038 137 0.00002 0.00103 0.00093 0.027 138 0.00003 0.00122 0.00258
0.020 139 0.00002 0.00124 0.00155 0.083 140 0.00002 0.00269 0.00134
0.100 141 0.00002 0.00117 0.00124 0.047 142 0.00002 0.00079 0.00087
0.078 143 0.00005 0.00302 0.00217 0.118 144 0.00002 0.00186 0.00123
0.056 145 0.00004 0.00090 0.00089 0.057 146 0.00002 0.00047 0.00039
0.018 147 0.00003 0.00169 0.00103 0.089 148 0.00002 0.00079 0.00055
0.091 149 0.00008 0.00888 0.00797 0.231 150 0.38044 >25 >25
>10 151 0.00004 0.00204 0.00218 0.113 152 0.05163 6.8 4.6 >10
153 0.00001 0.00074 0.00145 0.026 154 0.00005 0.00104 0.00074 0.257
155 0.00006 0.00220 0.00481 0.114 156 0.00004 0.00077 0.00080 0.178
157 0.00004 0.00120 0.00150 0.057 158 0.00004 0.00081 0.00132 0.040
159 0.00002 0.00109 0.00075 0.046 160 0.00005 0.00190 0.00222 0.082
161 0.00002 0.00087 0.00076 0.068 162 0.00168 0.059 163 0.00183
0.053 164 0.27024 >10 165 0.00002 0.00113 0.00095 0.066 166
0.00002 0.00040 0.00036 0.015 167 0.00018 0.00607 0.01257 0.240 168
0.00004 0.00531 0.00195 0.099 169 0.00079 0.06566 0.02227 2.363 170
0.00012 0.01384 0.00986 0.352 171 0.00002 0.01018 0.00251 0.398 172
0.00004 0.00169 0.00272 0.086 173 0.00005 0.00267 0.00132 0.136 174
0.00002 0.00065 0.00041 0.014 175 0.00009 0.00215 0.00252 0.091 176
0.00002 0.00077 0.00084 0.056 177 0.00004 0.00189 0.00296 0.069 178
0.00016 0.01494 0.02525 2.190 179 0.00003 0.00319 0.00516 0.266 180
0.00013 0.00747 0.00668 0.206 181 0.00003 0.00210 0.00175 0.108 182
0.00030 0.03743 0.02985 1.045 183 0.00001 0.00308 0.00075 0.058 184
0.00002 0.00204 0.00090 0.032 185 0.01333 0.417 186 0.00005 0.00405
0.00287 0.291 187 0.00004 0.00279 0.00203 0.163 188 0.00527 0.403
189 0.03192 4.4 1.3 >10 190 0.00087 0.04726 0.04313 >10 191
0.00019 0.02718 0.00971 0.747 192 0.00004 0.00442 0.00210 0.194 193
0.00003 0.00179 0.00062 0.092 194 0.00005 0.00154 0.00117 0.039 195
0.00010 0.01374 0.01423 0.528 196 0.00031 0.00260 0.00690 0.969 197
0.00007 0.00515 0.00209 0.400 198 0.00028 0.00218 0.00305 0.265 199
0.00005 0.00281 0.00356 0.098 200 0.00170 0.00519 0.01829 2.415 201
0.00017 0.02347 0.02474 0.424 202 0.00038 0.01614 0.01967 0.358 203
0.00005 0.00167 0.00152 0.059
[0573] Compound structures in the tables marked as "Chiral" were
prepared and tested in optically active form, having the absolute
stereochemistry as shown; other compounds were prepared and tested
in racemic form, and the depicted structure represents the relative
stereochemistry at each chiral center.
Sequence CWU 1
1
1115PRTartificial sequenceBad Peptide - biotinylated 1Ala Gly Ala
Gly Arg Ser Arg His Ser Ser Tyr Pro Ala Gly Thr 1 5 10 15
* * * * *