U.S. patent application number 14/287833 was filed with the patent office on 2014-11-27 for glucagon-like peptide 1 (glp-1) receptor modulators and uses thereof in regulating blood glucose levels.
This patent application is currently assigned to Academia Sinica. The applicant listed for this patent is Academia Sinica, Chi-Ming Liang. Invention is credited to Rong-Jie Chein, Klim King.
Application Number | 20140350100 14/287833 |
Document ID | / |
Family ID | 51935767 |
Filed Date | 2014-11-27 |
United States Patent
Application |
20140350100 |
Kind Code |
A1 |
King; Klim ; et al. |
November 27, 2014 |
GLUCAGON-LIKE PEPTIDE 1 (GLP-1) RECEPTOR MODULATORS AND USES
THEREOF IN REGULATING BLOOD GLUCOSE LEVELS
Abstract
The present disclosure provides novel glucagon-like peptide-1
(GLP-1) receptor modulators such as compounds of Formula (I) or
(II), and pharmaceutically acceptable salts thereof. The present
disclosure also provides pharmaceutical compositions, kits, and
uses that involve the GLP-1 receptor modulators for regulating
blood glucose levels and/or treating diabetes via, e.g., modulating
the endogenous signaling pathways mediated by the GLP-1 receptor.
##STR00001##
Inventors: |
King; Klim; (Taipei, TW)
; Chein; Rong-Jie; (Taipei, TW) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Liang; Chi-Ming
Academia Sinica |
Bethesda
Taipei |
MD |
US
TW |
|
|
Assignee: |
Academia Sinica
Taipei
MD
Liang; Chi-Ming
Bethesda
|
Family ID: |
51935767 |
Appl. No.: |
14/287833 |
Filed: |
May 27, 2014 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
61839870 |
Jun 27, 2013 |
|
|
|
61827674 |
May 27, 2013 |
|
|
|
Current U.S.
Class: |
514/510 ;
514/557; 514/691; 514/693; 514/729; 558/277; 560/117; 560/194;
560/220; 562/499; 568/373; 568/445; 568/817 |
Current CPC
Class: |
C07C 49/757 20130101;
C07C 67/30 20130101; C07C 69/96 20130101; C07C 323/15 20130101;
C07C 22/02 20130101; C07C 33/16 20130101; C07C 33/44 20130101; C07C
327/06 20130101; C07C 327/16 20130101; C07C 69/757 20130101; C07C
215/42 20130101; C07F 9/117 20130101; C07C 253/30 20130101; C07C
62/32 20130101; C07C 45/00 20130101; C07C 225/14 20130101; C07C
323/28 20130101; C07C 321/20 20130101; C07C 211/31 20130101; C07C
47/36 20130101; C07C 69/24 20130101; C07C 47/46 20130101; C07C
35/37 20130101; C07D 303/46 20130101; C07C 29/15 20130101; C07C
253/00 20130101; C07C 69/33 20130101; C07C 275/28 20130101; C07C
271/24 20130101; C07C 323/22 20130101; C07C 69/587 20130101; C07C
45/41 20130101; C07C 69/007 20130101; C07C 2603/30 20170501; A61P
3/10 20180101; C07C 259/06 20130101 |
Class at
Publication: |
514/510 ;
568/445; 514/693; 560/117; 568/817; 514/729; 568/373; 514/691;
562/499; 514/557; 560/220; 560/194; 558/277 |
International
Class: |
C07C 69/96 20060101
C07C069/96; C07C 69/757 20060101 C07C069/757; C07C 69/007 20060101
C07C069/007; C07C 49/757 20060101 C07C049/757; C07C 62/32 20060101
C07C062/32; C07C 47/36 20060101 C07C047/36; C07C 35/37 20060101
C07C035/37 |
Claims
1. A compound of Formula (I): ##STR00055## or a pharmaceutically
acceptable salt thereof, wherein: G.sub.A is hydrogen, .dbd.O,
.dbd.S, --OR'', --SR'', --N(R'').sub.2, alkenyl, alkynyl, an amide
group, an ester group, a phosphate group, an aldehyde group, a
nitrile group, an imino group, a ketone group, a thione group, an
isonitrile group, an isothiocyanide group, a carbamate group, a
thiocarbamate group, or a cyclic or acyclic, substituted or
unsubstituted, branched or unbranched, (hetero)aliphatic group
having 1 to 6 carbon atoms, wherein each instance of R'' is
independently hydrogen, a cyclic or acyclic, saturated or
unsaturated, substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 16 carbon atoms; R.sub.A1,
R.sub.A2, R.sub.A3, R.sub.A4, R.sub.A5, R.sub.A6, R.sub.A7,
R.sub.A8, R.sub.A9, and R.sub.A10 are each independently hydrogen,
halogen, --OR'', --N(R'').sub.2, a carboxyl group, or a cyclic or
acyclic, substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 6 carbons, or R.sub.A1 and
R.sub.A2 are joined to form .dbd.O, or R.sub.A3 and R.sub.A4 are
joined to form alkenyl; R.sub.A11, R.sub.A13, R.sub.A15, and
R.sub.A17 are each independently hydrogen, halogen, or a cyclic or
acyclic, substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 6 carbon atoms; R.sub.A12,
R.sub.A14, and R.sub.A16 are each independently halogen,
--N(R'').sub.2, --SR'', --OR'', alkyl, alkenyl, alkynyl, an amide
group, a carboxyl group, an ester group, an aldehyde group, a
nitrile group, an imino group, a ketone group, a thione group, an
isonitrile group, an isothiocyanide group, a urea group, a
carbamate group, or a thiocarbamate group, or R.sub.A14 and
R.sub.A15 are joined to form .dbd.O or .dbd.S; R.sub.A20 is
hydrogen, halogen, or a cyclic or acyclic, substituted or
unsubstituted, branched or unbranched, (hetero)aliphatic group
having 1 to 6 carbon atoms; and R.sub.A21 is hydrogen, halogen,
--N(R'').sub.2, --SR'', --OR'', --CH.sub.2OR'', alkenyl, alkynyl,
an amide group, a carboxyl group, an ester group, an aldehyde
group, a nitrile group, an imino group, a ketone group, a thione
group, an isonitrile group, an isothiocyanide group, a carbamate
group, a thiocarbamate group, or a cyclic or acyclic, substituted
or unsubstituted, branched or unbranched, (hetero)aliphatic group
having 1 to 6 carbon atoms; provided that: (i) at least one of
R.sub.A1, R.sub.A2, R.sub.A5, R.sub.A7, R.sub.A8, R.sub.A9,
R.sub.A10, R.sub.A11, R.sub.A12, R.sub.A13, R.sub.A15, R.sub.A16,
and R.sub.A17 is not hydrogen; (ii) at least one of R.sub.A3,
R.sub.A4, and R.sub.A6 is not --CH.sub.3; or (iii) when R.sub.A21
is --CHO and G.sub.A is --OH or .dbd.O, R.sub.A14 and R.sub.A15 are
each not --CHO.
2. The compound or pharmaceutically acceptable salt of claim 1,
wherein: G.sub.A is hydrogen, .dbd.O, .dbd.S, --OR'', --SR'',
--NR''H, alkenyl, alkynyl, an amide group, an ester group, an
aldehyde group, a nitrile group, an imino group, a ketone group, a
thione group, an isonitrile group, an isothiocyanide group, a
carbamate group, a thiocarbamate group, or a cyclic or acyclic,
substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 6 carbon atoms, wherein each
instance of R'' is independently hydrogen, a cyclic or acyclic,
saturated or unsaturated, substituted or unsubstituted, branched or
unbranched, (hetero)aliphatic group having 1 to 16 carbon atoms;
R.sub.A1, R.sub.A2, R.sub.A3, R.sub.A4, R.sub.A5, R.sub.A6,
R.sub.A7, R.sub.A8, R.sub.A9, and R.sub.A10 are each independently
hydrogen, halogen, or a cyclic or acyclic, substituted or
unsubstituted, branched or unbranched, (hetero)aliphatic group
having 1 to 6 carbons; R.sub.A11, R.sub.A12, R.sub.A13, R.sub.A15,
R.sub.A16, and R.sub.A17 are each independently hydrogen, halogen,
or a cyclic or acyclic, substituted or unsubstituted, branched or
unbranched, (hetero)aliphatic group having 1 to 6 carbon atoms;
R.sub.A14 is halogen, --NR''H, --SR'', --OR'', alkenyl, alkynyl, an
amide group, an ester group, an aldehyde group, a nitrile group, an
imino group, a ketone group, a thione group, an isonitrile group,
an isothiocyanide group, a carbamate group, or a thiocarbamate
group; R.sub.A20 is hydrogen, halogen, or a cyclic or acyclic,
substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 6 carbon atoms; and R.sub.A21
is hydrogen, halogen, --NR''H, --SR'', --OR'', alkenyl, alkynyl, an
amide group, an ester group, an aldehyde group, a nitrile group, an
imino group, a ketone group, a thione group, an isonitrile group,
an isothiocyanide group, a carbamate group, a thiocarbamate group,
or a cyclic or acyclic, substituted or unsubstituted, branched or
unbranched, (hetero)aliphatic group having 1 to 6 carbon atoms.
3. The compound or pharmaceutically acceptable salt of claim 1,
wherein the compound is of Formula (I-A), (I-A1), (I-B), or (I-C):
##STR00056##
4-6. (canceled)
7. The compound or pharmaceutically acceptable salt of claim 1,
wherein G.sub.A is .dbd.O, .dbd.S, --SR'', --OR'', --N(R'').sub.2,
--OH, --SH, or --NH.sub.2.
8. (canceled)
9. The compound or pharmaceutically acceptable salt of claim 1,
wherein R.sub.A6 is acyclic, substituted or unsubstituted, branched
or unbranched, C.sub.1-6 alkyl.
10. (canceled)
11. The compound or pharmaceutically acceptable salt of claim 1,
wherein R.sub.A14 is an ester group, an aldehyde group, a ketone
group, alkyl, a carboxyl group, or a urea group.
12. (canceled)
13. The compound or pharmaceutically acceptable salt of claim 1,
wherein R.sub.A21 is hydrogen, --CH.sub.2OR'', an aldehyde group,
or substituted or unsubstituted, branched or unbranched, C.sub.1-6
alkyl.
14-16. (canceled)
17. The compound or pharmaceutically acceptable salt of claim 1,
wherein the compound is of the formula: ##STR00057## ##STR00058##
##STR00059## ##STR00060## ##STR00061## ##STR00062## ##STR00063##
##STR00064## ##STR00065## ##STR00066## or a pharmaceutically
acceptable salt thereof, wherein ##STR00067##
18. A compound of Formula (II): ##STR00068## or a pharmaceutically
acceptable salt thereof, wherein: G is hydrogen, .dbd.O, .dbd.S,
--NR'H, --SR', or --OR', wherein R' is hydrogen, an ester group, a
ketone group, a thione group, or a cyclic or acyclic, saturated or
unsaturated, substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 16 carbon atoms; W is --O--,
--S-- or --NR'--; X and Y are each independently a single bond or a
saturated or unsaturated, substituted or unsubstituted, branched or
unbranched, (hetero)aliphatic group having 1 to 3 carbon atoms;
R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7,
R.sub.8, R.sub.9, R.sub.12 and R.sub.13 are each independently
hydrogen, halogen, or a cyclic or acyclic, substituted or
unsubstituted, branched or unbranched, (hetero)aliphatic group
having 1 to 6 carbon atoms, or R.sub.2 and R.sub.3 may join to form
cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; R.sub.10 and
R.sub.11 are each independently hydrogen, halogen, an amino group,
an amide group, an ester group, an aldehyde group, a nitrile, an
imino group, a ketone group, a thione group, an isonitrile group,
an isothiocyanide group, a carbamate group, a thiocarbamate group,
or a cyclic or acyclic, saturated or unsaturated, substituted or
unsubstituted, branched or unbranched, (hetero)aliphatic group
having 1 to 6 carbon atoms; R.sub.14 is hydrogen or a saturated or
unsaturated, substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1-16 carbon atoms; R.sub.15 is
hydrogen or a saturated or unsaturated, substituted or
unsubstituted, branched or unbranched, (hetero)aliphatic group
having 1-6 carbon atoms; and R.sub.21 is ##STR00069## or an
aldehyde group; provided that: (i) at least one of R.sub.4,
R.sub.5, R.sub.6, R.sub.7, R.sub.8, R.sub.9, and R.sub.10 is not
hydrogen; (ii) R.sub.1 is not --CH.sub.3; or (iii) when R.sub.21 is
--CHO and G is --OH or .dbd.O, R.sub.11 is not --CHO.
19. The compound or pharmaceutically acceptable salt of claim 18,
wherein the compound is of Formula (II-A), (II-B), or (II-C):
##STR00070##
20-21. (canceled)
22. The compound or pharmaceutically acceptable salt of claim 18,
wherein R.sub.21 is ##STR00071## --CH.sub.2OH, or an aldehyde
group.
23-24. (canceled)
25. The compound or pharmaceutically acceptable salt of claim 18,
wherein G is .dbd.O or --OR'.
26. (canceled)
27. The compound or pharmaceutically acceptable salt of claim 18,
wherein W is --O--, X is methylene, or Y is methylene.
28-29. (canceled)
30. The compound or pharmaceutically acceptable salt of claim 18,
wherein R.sub.11 is an ester group, an aldehyde group, a ketone
group, or acyclic, substituted or unsubstituted, branched or
unbranched, C.sub.1-6 alkyl.
31. A pharmaceutical composition comprising a compound or
pharmaceutically acceptable salt of claim 1, and a pharmaceutically
acceptable carrier.
32. A method for regulating blood glucose level in a subject,
comprising administering an effective amount of the pharmaceutical
composition of claim 31 to a subject in need thereof.
33. The method of claim 32, wherein the subject has, is suspected
of having, or is at risk a disease or disorder selected from the
group consisting of type I diabetes, type II diabetes, gestational
diabetes, obesity, excessive appetite, insufficient satiety, and a
metabolic disorder.
34. A method for activating a glucagon-like peptide 1 (GLP-1)
receptor in a subject in the presence of GLP-1, comprising
administering to a subject in need thereof an effective amount of
the pharmaceutical composition of claim 31.
35. A pharmaceutical composition comprising a compound or
pharmaceutically acceptable salt of claim 18, and a
pharmaceutically acceptable carrier.
36. A method for regulating blood glucose level or activating a
glucagon-like peptide 1 (GLP-1) receptor in a subject, the method
comprising administering to a subject in need thereof an effective
amount of the pharmaceutical composition of claim 35.
Description
RELATED APPLICATIONS
[0001] The present application claims priority under 35 U.S.C.
.sctn.119(e) to U.S. provisional patent applications, 61/827,674,
filed May 27, 2013 and 61/839,870, filed Jun. 27, 2013, the entire
content of each of which is incorporated by reference herein.
BACKGROUND
[0002] Type II diabetes is characterized by decrease in peripheral
tissue response to insulin in association with impaired cell
function, which results in increase in fasting glycemia (1,2).
Currently antihyperglycemic drugs such as metformin, sulfonylureas,
or thiazolidinediones have been prescribed to promote insulin
secretion or enhance insulin sensitivity, these drugs do not target
all of the symptoms of type II DM (3,4). In cretin hormones (e.g.,
glucagon-like peptide-1 (GLP-1) and gastric inhibitory polypeptide
GIP) are intestinally derived hormones that stimulate cAMP
production via their cognate receptors in pancreatic b cells and
subsequently leads to glucose dependent insulin secretion in
response to food intake, play an important role in glucose
homeostasis (5). Apart from its insulinotropic effects, GLP-1 also
preserves pancreatic cells, suppresses glucagon release, reduces
hepatic gluconeogenesis, it delays gastric emptying, reduces food
intake by promoting satiety and reveals a favorable cardiometabolic
profile (6-9). GLP-1 also displays extra-pancreatic effects,
notably targeting the brain, immune system and heart, where it
plays a role in neuroprotection (10-13), regulating immune
responses (14-16) and cardioprotection (17-19). All these
physiological actions of GLP-1 are mediated via interaction with
its cognate G-protein coupled receptor--GLP-1 receptor on the
target tissues. However, the exact downstream signaling of these
extra-pancreatic physiological actions of GLP-1 still needed
further investigation.
[0003] GPCR (G protein-coupled receptor) signaling machinery was
once considered as operating in an one-dimensional way, now has
been proved to signal through several distinct mechanisms including
those mediated by G proteins and by G-protein independent
multifunctional adaptor proteins .beta.-arrestins. In addition to
heterotrimeric G proteins, two protein families specifically
interact with the majority of GPCRs in their activated
conformation: G protein-coupled receptor kinases (GRKs) and
.beta.-arrestins (20). GRKs and .beta.-arrestins are considered to
be G protein-independent signal transducers (20-22). In particular,
(3-arrestins act as multifunctional scaffolds that interact with
many protein partners (23) and protein kinases, thereby leading to
the phosphorylation of numerous intracellular targets (24).
Distinct selective coupling pathway will elicit quite distinct
physiologic outcome and this finding has currently impacted further
on the development of assay technology and search for pathway
selective GPCR drug (25-28). Indeed, multi-pathway screening of
against u-opioid receptor (29) and agonist for parathyroid hormone
receptor (25) lead to discovery of compounds that provide
therapeutic effect without the adverse side effects normally
associated with these receptors, support the notion that pathway
selective (biased) agonists may identify new classes of therapeutic
agents that have fewer side effects. Though the insulinotropic
effect of GLP-1 is mediated by stimulating cAMP generation in
pancreatic cells via coupling to G.alpha.s, other effects of GLP-1
have been shown to be mediated via coupling to .beta.-arrestin.
GLP-1 anti-apoptotic effect on pancreatic .beta.-cells has been
shown to be mediated by phosphorylating the pro-apoptotic protein
Bad through .beta.-arrestin1 dependent ERK1/2 activation (30).
.beta.-Arrestin1-mediated recruitment of c-Src is involved in
proliferative action of GLP-1 on pancreatic .beta.-cells (31). It
will be interesting to know other extrapancreatic function of GLP-1
will be also mediated by arrestin coupling and it will be more
straightforward to use pathway selective compounds to correlate
cellular responses to specific signaling pathway. However, pathway
selective compound is rarely available for GLP-1 receptor
signaling.
[0004] Current GLP-1 analogue therapeutics requires frequent
subcutaneous administrations, and leads to reduced compliance and
high prices in developing area. Typically, the plasma level of
active GLP-1 is around 5 to 10 pM in the basal state, quickly rises
to 20 to 50 pM after oral glucose or meal and will slowly declines
to basal level over 2 hours (32-34). However, GLP-1 analogue
therapeutics usually require to maintain constantly a
supra-physiological level of GLP-1 analogues, thus lead to
activating GLP-1 receptors constitutively and may cause severe
complications upon chronic treatment (35-42).
[0005] Identification of novel compounds that modulate the
endogenous GLP-1 receptor signaling pathways can lead to the
development of new therapeutics useful in regulating blood glucose
levels, thereby treating diabetes or disorders associated with the
GLP-1 receptor.
SUMMARY
[0006] This present disclosure is based on the discovery of novel
GLP-1 receptor modulators (e.g., compounds of Formula (I) or (II),
as being capable of modulating the endogenous glucagon-like
peptide-1 (GLP-1) receptor signaling pathways and thus, may be
useful in regulating blood glucose levels and/or treating diabetes
and other disorders associated with the GLP-1 receptor.
Accordingly, the present disclosure features GLP-1 receptor
modulators such as compounds of Formula (I) or (II), or
pharmaceutically acceptable salts thereof, and methods of using
such compounds for regulating blood glucose levels and/or treating
diabetes.
[0007] In one aspect, the GLP-1 receptor modulators described
herein are compounds of Formula (I):
##STR00002##
and pharmaceutically acceptable salts thereof, wherein:
[0008] G.sub.A is hydrogen, .dbd.O, .dbd.S, --OR'', --SR'',
--N(R'').sub.2, alkenyl, alkynyl, an amide group, an ester group, a
phosphate group, an aldehyde group, a nitrile group, an imino
group, a ketone group, a thione group, an isonitrile group, an
isothiocyanide group, a carbamate group, a thiocarbamate group, or
a cyclic or acyclic, substituted or unsubstituted, branched or
unbranched, (hetero)aliphatic group having 1 to 6 carbon atoms,
wherein each instance of R'' is independently hydrogen, a cyclic or
acyclic, saturated or unsaturated, substituted or unsubstituted,
branched or unbranched, (hetero)aliphatic group having 1 to 16
carbon atoms;
[0009] R.sub.A1, R.sub.A2, R.sub.A3, R.sub.A4, R.sub.A5, R.sub.A6,
R.sub.A7, R.sub.A8, R.sub.A9, and R.sub.A10 are each independently
hydrogen, halogen, --OR'', --N(R'').sub.2, a carboxyl group, or a
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched, (hetero)aliphatic group having 1 to 6 carbons, or
R.sub.A1 and R.sub.A2 are joined to form .dbd.O, or R.sub.A3 and
R.sub.A4 are joined to form alkenyl;
[0010] R.sub.A11, R.sub.A13, R.sub.A15, and R.sub.A17 are each
independently hydrogen, halogen, or a cyclic or acyclic,
substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 6 carbon atoms;
[0011] R.sub.A12, R.sub.A14, and R.sub.A16 are each independently
halogen, --N(R'').sub.2, --SR'', --OR'', alkyl, alkenyl, alkynyl,
an amide group, a carboxyl group, an ester group, an aldehyde
group, a nitrile group, an imino group, a ketone group, a thione
group, an isonitrile group, an isothiocyanide group, a urea group,
a carbamate group, or a thiocarbamate group, or R.sub.A14 and
R.sub.A15 are joined to form .dbd.O or .dbd.S;
[0012] R.sub.A20 is hydrogen, halogen, or a cyclic or acyclic,
substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 6 carbon atoms; and
[0013] R.sub.A21 is hydrogen, halogen, --N(R'').sub.2, --SR'',
--OR'', --CH.sub.2OR'', alkenyl, alkynyl, an amide group, a
carboxyl group, an ester group, an aldehyde group, a nitrile group,
an imino group, a ketone group, a thione group, an isonitrile
group, an isothiocyanide group, a carbamate group, a thiocarbamate
group, or a cyclic or acyclic, substituted or unsubstituted,
branched or unbranched, (hetero)aliphatic group having 1 to 6
carbon atoms.
[0014] All compounds described herein include the compounds
themselves, as well as their salts and stereoisomers, if
applicable. The salts, for example, can be formed between a
positively charged substituent (e.g., amino) on a compound and an
anion. Suitable anions include, but are not limited to, chloride,
bromide, iodide, sulfate, nitrate, phosphate, citrate,
methanesulfonate, trifluoroacetate, and acetate. Likewise, a
negatively charged substituent (e.g., carboxylate) on a compound
can form a salt with a cation. Suitable cations include, but are
not limited to, sodium ion, potassium ion, magnesium ion, calcium
ion, and an ammonium cation such as teteramethylammonium ion.
[0015] In certain embodiments, a salt described herein is a
pharmaceutically acceptable salt. The term "pharmaceutically
acceptable salt" refers to those salts which are, within the scope
of sound medical judgment, suitable for use in contact with the
tissues of a subject (e.g., a human or non-human animal) without
undue toxicity, irritation, allergic response and the like, and are
commensurate with a reasonable benefit/risk ratio. Pharmaceutically
acceptable salts are well known in the art. For example, Berge et
al., describe pharmaceutically acceptable salts in detail in J.
Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by
reference. Pharmaceutically acceptable salts of the compounds
described herein include those derived from suitable inorganic and
organic acids and bases. In certain embodiments, a pharmaceutically
acceptable salt can be a salt described herein.
[0016] Compounds described herein can comprise one or more
asymmetric centers, and thus can exist in various isomeric forms,
e.g., enantiomers and/or diastereomers. For example, the compounds
described herein can be in the form of an individual enantiomer,
diastereomer or geometric isomer, or can be in the form of a
mixture of stereoisomers, including racemic mixtures and mixtures
enriched in one or more stereoisomer. Isomers can be isolated from
mixtures by methods known to those skilled in the art, including
chiral high pressure liquid chromatography (HPLC) and the formation
and crystallization of chiral salts; or preferred isomers can be
prepared by asymmetric syntheses. See, for example, Jacques et al.,
Enantiomers, Racemates and Resolutions (Wiley Interscience, New
York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel,
Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and
Wilen, Tables of Resolving Agents and Optical Resolutions p. 268
(E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind.
1972). The present disclosure additionally encompasses compounds
described herein as individual isomers substantially free of other
isomers, and alternatively, as mixtures of various isomers.
[0017] In a formula, is a single or double bond, and is absent (and
therefore any substituent attached thereto is also absent) or a
single bond.
[0018] Unless otherwise specified, a moiety described herein may be
unsubstituted or may be substituted (e.g., at least one hydrogen
atom of the moiety being replaced with a non-hydrogen atom or
group). When a group described herein is substituted, the group may
be substituted, as valency permits, with one or more substituents
independently selected from the group consisting of C.sub.1-6 alkyl
(e.g., unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, propyl,
or butyl) or substituted C.sub.1-6 alkyl (e.g., --CF.sub.3,
--CH.sub.2--CF.sub.3, or --C.sub.2F.sub.5)), --OR.sup.a1 (e.g.,
--OH, --OMe, or --OEt), --N(R.sup.a1).sub.2 (e.g., --NH.sub.2,
--NHMe, or --NMe.sub.2), --SR.sup.a1 (e.g., --SH or --SMe), .dbd.O,
.dbd.S, --CHO, --C(.dbd.O)N(R.sup.a1).sub.2 (e.g.,
--C(.dbd.O)NH.sub.2, --C(.dbd.O)NHMe, or --C(.dbd.O)NMe.sub.2),
--CN, --C(.dbd.O)OR.sup.a1 (e.g., --C(.dbd.O)OH, --C(.dbd.O)OMe, or
--C(.dbd.O)OEt), --OC(.dbd.O)R.sup.b1 (e.g., --OC(.dbd.O)Me,
--OC(.dbd.O)Et, or
--OC(.dbd.O)(CH.sub.2).sub.7CH.dbd.CHCH.sub.2CH.dbd.CH(CH.sub.2).sub.4CH.-
sub.3), --OC(.dbd.O)OR.sup.a1 (e.g., --OC(.dbd.O)OMe,
--OC(.dbd.O)OEt, or
--OC(.dbd.O)O(CH.sub.2).sub.7CH.dbd.CHCH.sub.2CH.dbd.CH(CH.sub.2).sub.4CH-
.sub.3)), --C(R.sup.b1).sub.2OR.sup.a1 (e.g., --CH.sub.2--OH or
--CH.sub.2--OMe), --C(R.sup.b1).sub.2SR.sup.a1 (e.g.,
--CH.sub.2--SH or --CH.sub.2--SMe),
--C(R.sup.b1).sub.2N(R.sup.a1).sub.2 (e.g., --CH.sub.2--NH.sub.2,
--CH.sub.2--NHMe, or --CH.sub.2--NMe.sub.2), and
--C(R.sup.b1).sub.2C(.dbd.O)OR.sup.a1 (e.g.,
--CH.sub.2--OC(.dbd.O)OMe, --CH.sub.2--OC(.dbd.O)OEt, or
--CH.sub.2--OC(.dbd.O)O(CH.sub.2).sub.7CH.dbd.CHCH.sub.2CH.dbd.CH(CH.sub.-
2).sub.4CH.sub.3)), wherein each instance of R.sup.a1 is
independently H, C.sub.1-6 alkyl (e.g., unsubstituted C.sub.1-6
alkyl (e.g., methyl, ethyl, propyl, or butyl) or substituted
C.sub.1-6 alkyl (e.g., --CF.sub.3, --CH.sub.2--CF.sub.3, or
--C.sub.2F.sub.5)), C.sub.2-6 alkenyl (e.g., unsubstituted
C.sub.2-6 alkenyl (e.g., vinyl)), 3- to 10-membered cycloalkyl
(e.g., unsubstituted 3- to 10-membered cycloalkyl (e.g.,
cyclopropyl)), or 6- to 10-membered aryl (e.g., phenyl (e.g.,
unsubstituted phenyl or substituted phenyl)), and each instance of
R.sup.b1 is independently H, halogen (e.g., F, Cl, Br, or I
(iodine)), C.sub.1-6 alkyl (e.g., unsubstituted C.sub.1-6 alkyl
(e.g., methyl, ethyl, propyl, or butyl) or substituted C.sub.1-6
alkyl (e.g., --CF.sub.3, --CH.sub.2--CF.sub.3, or
--C.sub.2F.sub.5)), C.sub.2-6 alkenyl (e.g., unsubstituted
C.sub.2-6 alkenyl (e.g., vinyl)), 3- to 10-membered cycloalkyl
(e.g., unsubstituted 3- to 10-membered cycloalkyl (e.g.,
cyclopropyl)), or 6- to 10-membered aryl (e.g., phenyl (e.g.,
unsubstituted phenyl or substituted phenyl)).
[0019] When a range of values is listed, it is intended to
encompass each value and sub-range within the range. For example
"C.sub.1-6 alkyl" is intended to encompass, C.sub.1, C.sub.2,
C.sub.3, C.sub.4, C.sub.5, C.sub.6, C.sub.1-6, C.sub.1-5,
C.sub.1-4, C.sub.1-3, C.sub.1-2, C.sub.2-6, C.sub.2-5, C.sub.2-4,
C.sub.2-3, C.sub.3-6, C.sub.3-5, C.sub.3-4, C.sub.4-6, C.sub.4-5,
and C.sub.5-6 alkyl.
[0020] The term "(hetero)aliphatic" refers to aliphatic or
heteroaliphatic. The term "aliphatic" refers to alkyl, alkenyl,
alkynyl, and carbocyclic groups. The term "heteroaliphatic" refers
to heteroalkyl, heteroalkenyl, heteroalkynyl, and heterocyclic
groups.
[0021] The term "alkyl" refers to a radical of a straight-chained
("unbranched") or branched, saturated, hydrocarbon group. In some
embodiments, an alkyl group has 1 to 6 carbon atoms ("C.sub.1-6
alkyl"). Examples of C.sub.1-6 alkyl groups include methyl
(C.sub.1), ethyl (C.sub.2), n-propyl (C.sub.3), isopropyl
(C.sub.3), n-butyl (C.sub.4), tert-butyl (C.sub.4), sec-butyl
(C.sub.4), iso-butyl (C.sub.4), n-pentyl (C.sub.5), 3-pentanyl
(C.sub.5), amyl (C.sub.5), neopentyl (C.sub.5), 3-methyl-2-butanyl
(C.sub.5), tertiary amyl (C.sub.5), and n-hexyl (C.sub.6).
[0022] The term "alkenyl" refers to a radical of a straight-chained
or branched hydrocarbon group having one or more carbon-carbon
double bonds (e.g., 1, 2, 3, or 4 double bonds). In some
embodiments, an alkenyl group has 2 to 6 carbon atoms ("C.sub.2-6
alkenyl"). The one or more carbon-carbon double bonds can be
internal (such as in 2-butenyl) or terminal (such as in 1-butenyl).
Examples of C.sub.2-6 alkenyl groups include ethenyl (C.sub.2),
1-propenyl (C.sub.3), 2-propenyl (C.sub.3), 1-butenyl (C.sub.4),
2-butenyl (C.sub.4), butadienyl (C.sub.4), pentenyl (C.sub.5),
pentadienyl (C.sub.5), and hexenyl (C.sub.6).
[0023] The term "alkynyl" refers to a radical of a straight-chained
or branched hydrocarbon group having one or more carbon-carbon
triple bonds (e.g., 1, 2, 3, or 4 triple bonds). In some
embodiments, an alkynyl group has 2 to 6 carbon atoms ("C.sub.2-6
alkynyl"). The one or more carbon-carbon triple bonds can be
internal (such as in 2-butynyl) or terminal (such as in 1-butynyl).
Examples of C.sub.2-6 alkynyl groups include ethynyl (C.sub.2),
1-propynyl (C.sub.3), 2-propynyl (C.sub.3), 1-butynyl (C.sub.4),
2-butynyl (C.sub.4), pentynyl (C.sub.5), and hexynyl (C.sub.6).
[0024] "Heteroalkyl" refers to an alkyl group as defined herein
which further includes at least one heteroatom (e.g., 1, 2, 3, or 4
heteroatoms) selected from oxygen, nitrogen, or sulfur within
(i.e., inserted between adjacent carbon atoms of) and/or placed at
one or more terminal position(s) of the parent chain. In some
embodiments, a heteroalkyl group is a saturated group having 1 to
16 carbon atoms and 1 or more heteroatoms within the parent chain
("heteroC.sub.1-16 alkyl"). In some embodiments, a heteroalkyl
group is a saturated group having 1 to 6 carbon atoms and 1 or more
heteroatoms within the parent chain ("heteroC.sub.1-6 alkyl"). In
some embodiments, a heteroalkyl group is a saturated group having 1
to 3 carbon atoms and 1 or more heteroatoms within the parent chain
("heteroC.sub.1-3 alkyl"). Unless otherwise specified, each
instance of a heteroalkyl group is independently unsubstituted or
substituted with one or more substituents.
[0025] "Heteroalkenyl" refers to an alkenyl group as defined herein
which further includes at least one heteroatom (e.g., 1, 2, 3, or 4
heteroatoms) selected from oxygen, nitrogen, or sulfur within
(i.e., inserted between adjacent carbon atoms of) and/or placed at
one or more terminal position(s) of the parent chain. In some
embodiments, a heteroalkenyl group has 2 to 16 carbon atoms, at
least one double bond, and 1 or more heteroatoms within the parent
chain ("heteroC.sub.2-16 alkenyl"). In some embodiments, a
heteroalkenyl group has 2 to 6 carbon atoms, at least one double
bond, and 1 or more heteroatoms within the parent chain
("heteroC.sub.2-6 alkenyl"). In some embodiments, a heteroalkenyl
group has 2 to 3 carbon atoms, at least one double bond, and 1 or
more heteroatoms within the parent chain ("heteroC.sub.2-3
alkenyl"). Unless otherwise specified, each instance of a
heteroalkenyl group is independently unsubstituted or substituted
with one or more substituents.
[0026] "Heteroalkynyl" refers to an alkynyl group as defined herein
which further includes at least one heteroatom (e.g., 1, 2, 3, or 4
heteroatoms) selected from oxygen, nitrogen, or sulfur within
(i.e., inserted between adjacent carbon atoms of) and/or placed at
one or more terminal position(s) of the parent chain. In some
embodiments, a heteroalkynyl group has 2 to 16 carbon atoms, at
least one triple bond, and 1 or more heteroatoms within the parent
chain ("heteroC.sub.2-16 alkynyl"). In some embodiments, a
heteroalkynyl group has 2 to 6 carbon atoms, at least one triple
bond, and 1 or more heteroatoms within the parent chain
("heteroC.sub.2-6 alkynyl"). In some embodiments, a heteroalkynyl
group has 2 to 3 carbon atoms, at least one triple bond, and 1 or
more heteroatoms within the parent chain ("heteroC.sub.2-3
alkynyl"). Unless otherwise specified, each instance of a
heteroalkynyl group is independently unsubstituted or substituted
with one or more substituents.
[0027] "Carbocyclyl," "carbocycle," or "carbocyclic" refers to a
radical of a non-aromatic cyclic hydrocarbon group having from 3 to
10 ring carbon atoms ("C.sub.3-10 carbocyclyl") and zero
heteroatoms in the non-aromatic ring system. In some embodiments, a
carbocyclyl group has 3 to 8 ring carbon atoms ("C.sub.3-8
carbocyclyl"). In some embodiments, a carbocyclyl group has 3 to 6
ring carbon atoms ("C.sub.3-6 carbocyclyl"). In some embodiments, a
carbocyclyl group has 3 to 6 ring carbon atoms ("C.sub.3-6
carbocyclyl"). In some embodiments, a carbocyclyl group has 5 to 10
ring carbon atoms ("C.sub.5-10 carbocyclyl"). Exemplary C.sub.3-6
carbocyclyl groups include, without limitation, cyclopropyl
(C.sub.3), cyclopropenyl (C.sub.3), cyclobutyl (C.sub.4),
cyclobutenyl (C.sub.4), cyclopentyl (C.sub.5), cyclopentenyl
(C.sub.5), cyclohexyl (C.sub.6), cyclohexenyl (C.sub.6),
cyclohexadienyl (C.sub.6), and the like. Exemplary C.sub.3-8
carbocyclyl groups include, without limitation, the aforementioned
C.sub.3-6 carbocyclyl groups as well as cycloheptyl (C.sub.7),
cycloheptenyl (C.sub.7), cycloheptadienyl (C.sub.7),
cycloheptatrienyl (C.sub.7), cyclooctyl (C.sub.8), cyclooctenyl
(C.sub.8), bicyclo[2.2.1]heptanyl (C.sub.7), bicyclo[2.2.2]octanyl
(C.sub.8), and the like. Exemplary C.sub.3-10 carbocyclyl groups
include, without limitation, the aforementioned C.sub.3-8
carbocyclyl groups as well as cyclononyl (C.sub.9), cyclononenyl
(C.sub.9), cyclodecyl (C.sub.1-10), cyclodecenyl (C.sub.1-10),
octahydro-1H-indenyl (C.sub.9), decahydronaphthalenyl (C.sub.1-10),
spiro[4.5]decanyl (C.sub.1-10), and the like. As the foregoing
examples illustrate, in certain embodiments, the carbocyclyl group
is either monocyclic ("monocyclic carbocyclyl") or contain a fused,
bridged. or spiro ring system such as a bicyclic system ("bicyclic
carbocyclyl"). Carbocyclyl can be saturated, and saturated
carbocyclyl is referred to as "cycloalkyl." In some embodiments,
carbocyclyl is a monocyclic, saturated carbocyclyl group having
from 3 to 10 ring carbon atoms ("C.sub.3-10 cycloalkyl"). In some
embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms
("C.sub.3-8 cycloalkyl"). In some embodiments, a cycloalkyl group
has 3 to 6 ring carbon atoms ("C.sub.3-4 cycloalkyl"). In some
embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms
("C.sub.5-6 cycloalkyl"). In some embodiments, a cycloalkyl group
has 5 to 10 ring carbon atoms ("C.sub.5-10 cycloalkyl"). Examples
of C.sub.5-4 cycloalkyl groups include cyclopentyl (C.sub.5) and
cyclohexyl (C.sub.5). Examples of C.sub.3-4 cycloalkyl groups
include the aforementioned C.sub.54cycloalkyl groups as well as
cyclopropyl (C.sub.3) and cyclobutyl (C.sub.4). Examples of
C.sub.3-8 cycloalkyl groups include the aforementioned C.sub.3-4
cycloalkyl groups as well as cycloheptyl (C.sub.7) and cyclooctyl
(C.sub.8). Unless otherwise specified, each instance of a
cycloalkyl group is independently unsubstituted (an "unsubstituted
cycloalkyl") or substituted (a "substituted cycloalkyl") with one
or more substituents. In certain embodiments, the cycloalkyl group
is unsubstituted C.sub.3-10 cycloalkyl. In certain embodiments, the
cycloalkyl group is substituted C.sub.3-10 cycloalkyl. Carbocyclyl
can be partially unsaturated. Carbocyclyl including one or more
C.dbd.C double bond in the carbocyclic ring is referred to as
"cycloalkenyl." Carbocyclyl including one or more C.ident.C triple
bond in the carbocyclic ring is referred to as "cycloalkynyl."
Carbocyclyl includes aryl. "Carbocyclyl" also includes ring systems
wherein the carbocyclic ring, as defined above, is fused with one
or more aryl or heteroaryl groups wherein the point of attachment
is on the carbocyclic ring, and in such instances, the number of
carbons continue to designate the number of carbons in the
carbocyclic ring system. Unless otherwise specified, each instance
of a carbocyclyl group is independently optionally substituted,
i.e., unsubstituted (an "unsubstituted carbocyclyl") or substituted
(a "substituted carbocyclyl") with one or more substituents. In
certain embodiments, the carbocyclyl group is unsubstituted
C.sub.3-10 carbocyclyl. In certain embodiments, the carbocyclyl
group is substituted C.sub.3-10 carbocyclyl.
[0028] "Heterocyclyl," "heterocycle," or "heterocyclic" refers to a
radical of a 3- to 10-membered non-aromatic ring system having ring
carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom
is independently selected from nitrogen, oxygen, sulfur, boron,
phosphorus, and silicon ("3-10 membered heterocyclyl"). In
heterocyclyl groups that contain one or more nitrogen atoms, the
point of attachment can be a carbon or nitrogen atom, as valency
permits. A heterocyclyl group can either be monocyclic ("monocyclic
heterocyclyl") or a fused, bridged, or spiro ring system, such as a
bicyclic system ("bicyclic heterocyclyl"), and can be saturated
("heterocycloalkyl") or can be partially unsaturated. Heterocyclyl
bicyclic ring systems can include one or more heteroatoms in one or
both rings. Heterocyclyl includes heteroaryl. Heterocyclyl also
includes ring systems wherein the heterocyclic ring, as defined
above, is fused with one or more carbocyclyl groups wherein the
point of attachment is either on the carbocyclyl or heterocyclic
ring, or ring systems wherein the heterocyclic ring, as defined
above, is fused with one or more aryl or heteroaryl groups, wherein
the point of attachment is on the heterocyclic ring, and in such
instances, the number of ring members continue to designate the
number of ring members in the heterocyclic ring system. Unless
otherwise specified, each instance of heterocyclyl is independently
optionally substituted, i.e., unsubstituted (an "unsubstituted
heterocyclyl") or substituted (a "substituted heterocyclyl") with
one or more substituents. In certain embodiments, the heterocyclyl
group is unsubstituted 3-10 membered heterocyclyl. In certain
embodiments, the heterocyclyl group is substituted 3-10 membered
heterocyclyl.
[0029] In some embodiments, a heterocyclyl group is a 5-10 membered
non-aromatic ring system having ring carbon atoms and 1-4 ring
heteroatoms, wherein each heteroatom is independently selected from
nitrogen, oxygen, sulfur, boron, phosphorus, and silicon ("5-10
membered heterocyclyl"). In some embodiments, a heterocyclyl group
is a 5-8 membered non-aromatic ring system having ring carbon atoms
and 1-4 ring heteroatoms, wherein each heteroatom is independently
selected from nitrogen, oxygen, and sulfur ("5-8 membered
heterocyclyl"). In some embodiments, a heterocyclyl group is a 5-6
membered non-aromatic ring system having ring carbon atoms and 1-4
ring heteroatoms, wherein each heteroatom is independently selected
from nitrogen, oxygen, and sulfur ("5-6 membered heterocyclyl"). In
some embodiments, the 5-6 membered heterocyclyl has 1-3 ring
heteroatoms selected from nitrogen, oxygen, and sulfur. In some
embodiments, the 5-6 membered heterocyclyl has 1-2 ring heteroatoms
selected from nitrogen, oxygen, and sulfur. In some embodiments,
the 5-6 membered heterocyclyl has one ring heteroatom selected from
nitrogen, oxygen, and sulfur.
[0030] Exemplary 3-membered heterocyclyl groups containing one
heteroatom include, without limitation, azirdinyl, oxiranyl, and
thiiranyl. Exemplary 4-membered heterocyclyl groups containing one
heteroatom include, without limitation, azetidinyl, oxetanyl and
thietanyl. Exemplary 5-membered heterocyclyl groups containing one
heteroatom include, without limitation, tetrahydrofuranyl,
dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl,
pyrrolidinyl, dihydropyrrolyl and pyrrolyl-2,5-dione. Exemplary
5-membered heterocyclyl groups containing two heteroatoms include,
without limitation, dioxolanyl, oxasulfuranyl, disulfuranyl, and
oxazolidin-2-one. Exemplary 5-membered heterocyclyl groups
containing three heteroatoms include, without limitation,
triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary
6-membered heterocyclyl groups containing one heteroatom include,
without limitation, piperidinyl, tetrahydropyranyl,
dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclyl
groups containing two heteroatoms include, without limitation,
piperazinyl, morpholinyl, dithianyl, and dioxanyl. Exemplary
6-membered heterocyclyl groups containing three heteroatoms
include, without limitation, triazinanyl. Exemplary 7-membered
heterocyclyl groups containing one heteroatom include, without
limitation, azepanyl, oxepanyl and thiepanyl. Exemplary 8-membered
heterocyclyl groups containing one heteroatom include, without
limitation, azocanyl, oxecanyl and thiocanyl. Exemplary 5-membered
heterocyclyl groups fused to a C.sub.6 aryl ring (also referred to
herein as a 5,6-bicyclic heterocyclic ring) include, without
limitation, indolinyl, isoindolinyl, dihydrobenzofuranyl,
dihydrobenzothienyl, benzoxazolinonyl, and the like. Exemplary
6-membered heterocyclyl groups fused to an aryl ring (also referred
to herein as a 6,6-bicyclic heterocyclic ring) include, without
limitation, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the
like.
[0031] "Aryl" refers to a radical of a monocyclic or polycyclic
(e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g.,
having 6, 10, or 14 pi electrons shared in a cyclic array) having
6-14 ring carbon atoms and zero heteroatoms provided in the
aromatic ring system ("C.sub.6-14 aryl"). In some embodiments, an
aryl group has six ring carbon atoms ("C.sub.6 aryl"; e.g.,
phenyl). In some embodiments, an aryl group has ten ring carbon
atoms ("C.sub.10 aryl"; e.g., naphthyl such as 1-naphthyl and
2-naphthyl). In some embodiments, an aryl group has fourteen ring
carbon atoms ("C.sub.14 aryl"; e.g., anthracyl). "Aryl" also
includes ring systems wherein the aryl ring, as defined above, is
fused with one or more carbocyclyl or heterocyclyl groups wherein
the radical or point of attachment is on the aryl ring, and in such
instances, the number of carbon atoms continue to designate the
number of carbon atoms in the aryl ring system. Unless otherwise
specified, each instance of an aryl group is independently
optionally substituted, i.e., unsubstituted (an "unsubstituted
aryl") or substituted (a "substituted aryl") with one or more
substituents. In certain embodiments, the aryl group is
unsubstituted C.sub.6-14 aryl. In certain embodiments, the aryl
group is substituted C.sub.6-14 aryl.
[0032] "Aralkyl" is a subset of alkyl and aryl, as defined herein,
and refers to an optionally substituted alkyl group substituted by
an optionally substituted aryl group. In certain embodiments, the
aralkyl is optionally substituted benzyl. In certain embodiments,
the aralkyl is benzyl. In certain embodiments, the aralkyl is
optionally substituted phenethyl. In certain embodiments, the
aralkyl is phenethyl.
[0033] "Aralkenyl" is a subset of alkenyl and aryl, as defined
herein, and refers to an optionally substituted alkenyl group
substituted by an optionally substituted aryl group. An example of
aralkenyl is styrenyl (i.e., --CH.dbd.CHPh).
[0034] "Aralkynyl" is a subset of alkynyl and aryl, as defined
herein, and refers to an optionally substituted alkynyl group
substituted by an optionally substituted aryl group.
[0035] "Heteroaryl" refers to a radical of a 5-10 membered
monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or
10 pi electrons shared in a cyclic array) having ring carbon atoms
and 1-4 ring heteroatoms provided in the aromatic ring system,
wherein each heteroatom is independently selected from nitrogen,
oxygen and sulfur ("5-10 membered heteroaryl"). In heteroaryl
groups that contain one or more nitrogen atoms, the point of
attachment can be a carbon or nitrogen atom, as valency permits.
Heteroaryl bicyclic ring systems can include one or more
heteroatoms in one or both rings. "Heteroaryl" includes ring
systems wherein the heteroaryl ring, as defined above, is fused
with one or more carbocyclyl or heterocyclyl groups wherein the
point of attachment is on the heteroaryl ring, and in such
instances, the number of ring members continue to designate the
number of ring members in the heteroaryl ring system. "Heteroaryl"
also includes ring systems wherein the heteroaryl ring, as defined
above, is fused with one or more aryl groups wherein the point of
attachment is either on the aryl or heteroaryl ring, and in such
instances, the number of ring members designates the number of ring
members in the fused (aryl/heteroaryl) ring system. Bicyclic
heteroaryl groups wherein one ring does not contain a heteroatom
(e.g., indolyl, quinolinyl, carbazolyl, and the like) the point of
attachment can be on either ring, i.e., either the ring bearing a
heteroatom (e.g., 2-indolyl) or the ring that does not contain a
heteroatom (e.g., 5-indolyl).
[0036] In some embodiments, a heteroaryl group is a 5-10 membered
aromatic ring system having ring carbon atoms and 1-4 ring
heteroatoms provided in the aromatic ring system, wherein each
heteroatom is independently selected from nitrogen, oxygen, and
sulfur ("5-10 membered heteroaryl"). In some embodiments, a
heteroaryl group is a 5-8 membered aromatic ring system having ring
carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring
system, wherein each heteroatom is independently selected from
nitrogen, oxygen, and sulfur ("5-8 membered heteroaryl"). In some
embodiments, a heteroaryl group is a 5-6 membered aromatic ring
system having ring carbon atoms and 1-4 ring heteroatoms provided
in the aromatic ring system, wherein each heteroatom is
independently selected from nitrogen, oxygen, and sulfur ("5-6
membered heteroaryl"). In some embodiments, the 5-6 membered
heteroaryl has 1-3 ring heteroatoms selected from nitrogen, oxygen,
and sulfur. In some embodiments, the 5-6 membered heteroaryl has
1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In
some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom
selected from nitrogen, oxygen, and sulfur. Unless otherwise
specified, each instance of a heteroaryl group is independently
optionally substituted, i.e., unsubstituted (an "unsubstituted
heteroaryl") or substituted (a "substituted heteroaryl") with one
or more substituents. In certain embodiments, the heteroaryl group
is unsubstituted 5-14 membered heteroaryl. In certain embodiments,
the heteroaryl group is substituted 5-14 membered heteroaryl.
[0037] Exemplary 5-membered heteroaryl groups containing one
heteroatom include, without limitation, pyrrolyl, furanyl and
thiophenyl. Exemplary 5-membered heteroaryl groups containing two
heteroatoms include, without limitation, imidazolyl, pyrazolyl,
oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary
5-membered heteroaryl groups containing three heteroatoms include,
without limitation, triazolyl, oxadiazolyl, and thiadiazolyl.
Exemplary 5-membered heteroaryl groups containing four heteroatoms
include, without limitation, tetrazolyl. Exemplary 6-membered
heteroaryl groups containing one heteroatom include, without
limitation, pyridinyl. Exemplary 6-membered heteroaryl groups
containing two heteroatoms include, without limitation,
pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered
heteroaryl groups containing three or four heteroatoms include,
without limitation, triazinyl and tetrazinyl, respectively.
Exemplary 7-membered heteroaryl groups containing one heteroatom
include, without limitation, azepinyl, oxepinyl, and thiepinyl.
Exemplary 5,6-bicyclic heteroaryl groups include, without
limitation, indolyl, isoindolyl, indazolyl, benzotriazolyl,
benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl,
benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl,
benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and
purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, without
limitation, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl,
cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.
[0038] The term "oxo" refers to the a moiety of the formula:
.dbd.O.
[0039] The term "amide" or "amide group" refers to a moiety of the
formula: --N(R.sup.pp)C(.dbd.O)R.sup.qq, wherein R.sup.pp is a
nitrogen atom substituent described herein, and R.sup.qq is a
carbon atom substituent described herein.
[0040] The term "ester" or "ester group" refers to a moiety of the
formula: --C(.dbd.O)OR.sup.rr or --OC(.dbd.O)R.sup.rr, wherein
R.sup.rr is an oxygen atom substituent described herein.
[0041] The term "phosphate" or "phosphate group" refers to a moiety
of the formula: --OP(.dbd.O)(OR.sup.oo).sub.2, wherein each
instance of R.sup.oo is independently an oxygen atom substituent
described herein or a cationic counterion.
[0042] The term "carboxyl" or "carboxyl group" refers to a moiety
of the formula: --C(.dbd.O)OH.
[0043] The term "aldehyde" or "aldehyde group" refers to a moiety
of the formula: --C(.dbd.O)H.
[0044] The term "thialdehyde" or "thialdehyde group" refers to a
moiety of the formula: --C(.dbd.S)H.
[0045] The term "nitrile" or "nitrile group" refers to a moiety of
the formula: --CN or -L-CN, wherein L is substituted or
unsubstituted, branched or unbranched, C.sub.1-16 alkylene;
substituted or unsubstituted, branched or unbranched, C.sub.2-16
alkenylene; or substituted or unsubstituted, branched or
unbranched, C.sub.2-16 alkynylene.
[0046] The term "alcohol," "alcohol group," "hydroxyl," or
"hydroxy" refers to the group --OH. The term "substituted hydroxyl"
or "substituted hydroxyl" refers to a hydroxyl group wherein the
oxygen atom directly attached to the parent molecule is substituted
with a group other than hydrogen, and includes groups selected from
--OR.sup.aa, --ON(R.sup.bb).sub.2, --OC(.dbd.O)SR.sup.aa,
--OC(.dbd.O)R.sup.aa, --OCO.sub.2R.sup.aa,
--OC(.dbd.O)N(R.sup.bb).sub.2, --OC(.dbd.NR.sup.bb)R.sup.aa,
--OC(.dbd.NR.sup.bb)OR.sup.aa,
--OC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2, --OS(.dbd.O)R.sup.aa,
--OSO.sub.2R.sup.aa, --OSi(R.sup.aa).sub.3, --OP(R.sup.cc).sub.2,
--OP(R.sup.cc).sub.3, --OP(.dbd.O).sub.2R.sup.aa,
--OP(.dbd.O)(R.sup.aa).sub.2, --OP(.dbd.O)(OR.sup.cc).sub.2,
--OP(.dbd.O).sub.2N(R.sup.bb).sub.2, and
--OP(.dbd.O)(NR.sup.bb).sub.2, wherein R.sup.aa, R.sup.bb, and
R.sup.cc are as defined herein.
[0047] The term "amino" or "amino group" refers to a moiety of the
formula: --N(R.sup.ii).sub.2, wherein each instance of R.sup.ii is
independently a nitrogen atom substituent described herein, or two
instances of R.sup.ii are connected to form substituted or
unsubstituted heterocyclyl. In certain embodiments, the amino is
unsubstituted amino (i.e., --NH.sub.2). In certain embodiments, the
amino is a substituted amino group, wherein at least one instance
of R.sup.ii is not hydrogen.
[0048] The term "imino" or "imino group" refers to a moiety of the
formula: .dbd.NR.sup.ss, wherein R.sup.ss is a nitrogen atom
substituent described herein.
[0049] The term "ketone" or "ketone group" refers to a moiety of
the formula: --C(.dbd.O)R.sup.tt, wherein R.sup.tt is a carbon atom
substituent described herein.
[0050] The term "thione" or "thione group" refers to a moiety of
the formula: --C(.dbd.S)R.sup.uu, wherein R.sup.uu is a carbon atom
substituent described herein.
[0051] The term "isonitrile" or "isonitrile group" refers to a
moiety of the formula: --NC.
[0052] The term "isothiocyanide" or "isothiocyanide group" refers
to a moiety of the formula: --SNC.
[0053] The term "thioate" or "thioate group" refers to a moiety of
the formula: --C(.dbd.O)SR.sup.zz or --C(.dbd.S)OR.sup.jj, wherein
R.sup.zz is a sulfur atom substituent described herein, and R is an
oxygen atom substituent described herein.
[0054] The term "thioamide" or "thioamide group" refers to a moiety
of the formula: --N(R.sup.mm)C(.dbd.S)R.sup.nn, wherein R.sup.mm is
a nitrogen atom substituent described herein, and R.sup.nn is a
carbon atom substituent described herein.
[0055] The term "dithioate" or "dithioate group" refers to a moiety
of the formula: --C(.dbd.S)SR.sup.kk, wherein R.sup.kk is a sulfur
atom substituent described herein.
[0056] The term "isocyanato" or "isocyanato group" refers to a
moiety of the formula: --NCO.
[0057] The term "isothiocyanato" or "isothiocyanato group" refers
to a moiety of the formula: --NCS.
[0058] The term "carbamate" or "carbamate group" refers to a moiety
of the formula: --N(R.sup.vv)C(.dbd.O)OR.sup.ww or
--OC(.dbd.O)N(R.sup.vv).sub.2, wherein each instance of R.sup.vv is
independently a nitrogen atom substituent described herein, and
R.sup.ww is an oxygen atom substituent described herein.
[0059] The term "urea" or "urea group" refers to a moiety of the
formula: --N(R.sup.zl)C(.dbd.O)N(R.sup.zl).sub.2, wherein each
instance of R.sup.zl is independently a nitrogen atom substituent
described herein.
[0060] The term "thiocarbamate" or "thiocarbamate group" refers to
a moiety of the formula: --N(R.sup.vv)C(.dbd.S)OR.sup.ww or
--OC(.dbd.S)N(R.sup.vv).sub.2, --N(R.sup.vv)C(.dbd.O)SR.sup.yy or
--SC(.dbd.O)N(R.sup.vv).sub.2, wherein each instance of R.sup.vv is
independently a nitrogen atom substituent described herein,
R.sup.ww is an oxygen atom substituent described herein, and
R.sup.yy is a sulfur atom substituent described herein.
[0061] "Halo" or "halogen" refers to fluorine (fluoro, F), chlorine
(chloro or Cl), bromine (bromo or Br), or iodine (iodo or I).
[0062] An atom, moiety, or group described herein may be
unsubstituted or substituted, as valency permits, unless otherwise
expressly provided. The term "substituted" refers to that at least
one hydrogen present on a group (e.g., a carbon or nitrogen atom)
is replaced with a permissible substituent, e.g., a substituent
which upon substitution results in a stable compound, e.g., a
compound which does not spontaneously undergo transformation such
as by rearrangement, cyclization, elimination, or other reaction.
Unless otherwise indicated, a "substituted" group has a substituent
at one or more substitutable positions of the group, and when more
than one position in any given structure is substituted, the
substituent is either the same or different at each position. The
term "substituted" is contemplated to include substitution with all
permissible substituents of organic compounds, any of the
substituents described herein that results in the formation of a
stable compound. The present disclosure contemplates any and all
such combinations in order to arrive at a stable compound. For
purposes of this disclosure, heteroatoms such as nitrogen may have
hydrogen substituents and/or any suitable substituent as described
herein which satisfy the valencies of the heteroatoms and results
in the formation of a stable moiety. In certain embodiments, the
substituent is a carbon atom substituent. In certain embodiments,
the substituent is a nitrogen atom substituent. In certain
embodiments, the substituent is an oxygen atom substituent. In
certain embodiments, the substituent is a sulfur atom substituent.
In certain embodiments, a substituent may contribute to optical
isomerism and/or stereo isomerism of a compound.
[0063] Exemplary carbon atom substituents include, but are not
limited to, halogen, --CN, --NO.sub.2, --N.sub.3, --SO.sub.2H,
--SO.sub.3H, --OH, --OR.sup.aa, --ON(R.sup.bb).sub.2,
--N(R.sup.bb).sub.2, --N(R.sup.bb).sub.3.sup.+X.sup.-,
--N(OR.sup.cc)R.sup.bb, --SH, --SR.sup.aa, --SSR.sup.cc,
--C(.dbd.O)R.sup.aa, --CO.sub.2H, --CHO, --C(OR.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --OC(.dbd.O)R.sup.aa, --OCO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.2, --OC(.dbd.O)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.O)R.sup.aa, --NR.sup.bbCO.sub.2R.sup.aa,
--NR.sup.bbC(.dbd.O)N(R.sup.bb).sub.2, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --OC(.dbd.NR.sup.bb)R.sup.aa,
--OC(.dbd.NR.sup.bb)OR.sup.aa,
--C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--OC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--C(.dbd.O)NR.sup.bbSO.sub.2R.sup.aa, --NR.sup.bbSO.sub.2R--,
--SO.sub.2N(R.sup.bb).sub.2, --SO.sub.2R.sup.aa,
--SO.sub.200R.sup.aa, --OSO.sub.2R.sup.aa, --S(.dbd.O)R.sup.aa,
--OS(.dbd.O)R.sup.aa, --Si(R.sup.aa).sub.3,
--OSi(R.sup.aa).sub.3--C(.dbd.S)N(R.sup.bb).sub.2,
--C(.dbd.O)SR.sup.aa, --C(.dbd.S)SR.sup.aa, --SC(.dbd.S)SR.sup.aa,
--SC(.dbd.O)SR.sup.aa, --OC(.dbd.O)SR.sup.aa,
--SC(.dbd.O)OR.sup.aa, --SC(.dbd.O)R.sup.aa,
--P(.dbd.O).sub.2R.sup.aa, --OP(.dbd.O).sub.2R.sup.aa,
--P(.dbd.O)(R.sup.aa).sub.2, --OP(.dbd.O)(R.sup.aa).sub.2,
--OP(.dbd.O)(OR.sup.cc).sub.2, --P(.dbd.O).sub.2N(R.sup.bb).sub.2,
--OP(.dbd.O).sub.2N(R.sup.bb).sub.2, --P(.dbd.O)(NR.sup.bb).sub.2,
--OP(.dbd.O)(NR.sup.bb).sub.2,
--NR.sup.bbP(.dbd.O)(OR.sup.cc).sub.2,
--NR.sup.bbP(.dbd.O)(NR.sup.bb).sub.2, --P(R.sup.cc).sub.2,
--P(R.sup.cc).sub.3, --OP(R.sup.cc).sub.2, --OP(R.sup.cc).sub.3,
--B(R.sup.aa).sub.2, --B(OR.sup.cc).sub.2, --BR.sup.aa(OR.sup.cc),
C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl,
wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 R.sup.dd groups; or two geminal hydrogens on a carbon atom
are replaced with the group .dbd.O, .dbd.S,
.dbd.NN(R.sup.bb).sub.2, .dbd.NNR.sup.bbC(.dbd.O)R.sup.aa,
.dbd.NNR.sup.bbC(.dbd.O)OR.sup.aa,
.dbd.NNR.sup.bbS(.dbd.O).sub.2R.sup.aa, .dbd.NR.sup.bb, or
.dbd.NOR.sup.cc,
[0064] each instance of R.sup.aa is, independently, selected from
C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl, or two
R.sup.aa groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd
groups;
[0065] each instance of R.sup.bb is, independently, selected from
hydrogen, --OH, --OR.sup.aa, --N(R.sup.cc).sub.2, --CN,
--C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.cc)OR.sup.aa, --C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2,
--SO.sub.2N(R.sup.cc).sub.2, --SO.sub.2R.sup.cc,
--SO.sub.2OR.sup.cc, --SOR.sup.aa, --C(.dbd.S)N(R.sup.cc).sub.2,
--C(.dbd.O)SR.sup.cc, --C(.dbd.S)SR.sup.cc,
--P(.dbd.O).sub.2R.sup.aa, --P(.dbd.O)(R.sup.aa).sub.2,
--P(.dbd.O).sub.2N(R.sup.cc).sub.2, --P(.dbd.O)(NR.sup.cc).sub.2,
C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl, or two
R.sup.bb groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd
groups;
[0066] each instance of R.sup.cc is, independently, selected from
hydrogen, C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10
alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl, or two
R.sup.cc groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups;
each instance of R.sup.dd is, independently, selected from halogen,
--CN, --NO.sub.2, --N.sub.3, --SO.sub.2H, --SO.sub.3H, --OH,
--OR.sup.ee, --ON(R.sup.ff).sub.2, --N(R.sup.ff).sub.2,
--N(R.sup.ff).sub.3.times., --N(OR.sup.ee)R.sup.ff, --SH,
--SR.sup.ee, --SSR.sup.ee, --C(.dbd.O)R.sup.ee, --CO.sub.2H,
--CO.sub.2R.sup.ee, --OC(.dbd.O)R.sup.ee, --OCO.sub.2R.sup.ee,
--C(.dbd.O)N(R.sup.ff).sub.2, --OC(.dbd.O)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.O)R.sup.ee, --NR.sup.ffCO.sub.2R.sup.ee,
--NR.sup.ffC(.dbd.O)N(R.sup.ff).sub.2,
--C(.dbd.NR.sup.ff)OR.sup.ee, --OC(.dbd.NR.sup.ff)R.sup.ee,
--OC(.dbd.NR.sup.ff)OR.sup.ee,
--C(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--OC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffSO.sub.2R.sup.ee, --SO.sub.2N(R.sup.ff).sub.2,
--SO.sub.2R.sup.ee, --SO.sub.2OR.sup.ee, --OSO.sub.2R.sup.ee,
--S(.dbd.O)R.sup.ee, --Si(R.sup.ee).sub.3, --OSi(R.sup.ee).sub.3,
--C(.dbd.S)N(R.sup.ff).sub.2, --C(.dbd.O)SR.sup.ee,
--C(.dbd.S)SR.sup.ee, --SC(.dbd.S)SR.sup.ee,
--P(.dbd.O).sub.2R.sup.ee, --P(.dbd.O)(R.sup.ee).sub.2,
--OP(.dbd.O)(R.sup.ee).sub.2, --OP(.dbd.O)(OR.sup.ee).sub.2,
C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered
heterocyclyl, C.sub.6-10 aryl, 5-10 membered heteroaryl, wherein
each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and
heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5
R.sup.gg groups, or two geminal R.sup.dd substituents can be joined
to form .dbd.O or .dbd.S;
[0067] each instance of R.sup.ee is, independently, selected from
C.sub.1-6 alkyl, C.sub.14 perhaloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, C.sub.6-10 aryl, 3-10
membered heterocyclyl, and 3-10 membered heteroaryl, wherein each
alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and
heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5
R.sup.gg groups;
[0068] each instance of R.sup.ff is, independently, selected from
hydrogen, C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6
alkenyl, C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered
heterocyclyl, C.sub.6-10 aryl and 5-10 membered heteroaryl, or two
R.sup.ff groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups;
and
[0069] each instance of R.sup.gg is, independently, halogen, --CN,
--NO.sub.2, --N.sub.3, --SO.sub.2H, --SO.sub.3H, --OH, --OC.sub.1-6
alkyl, --ON(C.sub.1-6 alkyl).sub.2, --N(C.sub.1-6 alkyl).sub.2,
--N(C.sub.1-6 alkyl).sub.3.sup.+X.sup.-, --NH(C.sub.1-6
alkyl).sub.2.sup.+X.sup.-, --NH.sub.2(C.sub.1-6
alkyl).sup.+X.sup.-, --NH.sub.3.sup.+X.sup.-, --N(OC.sub.1
alkyl)(C.sub.1-6 alkyl), --N(OH)(C.sub.1-6 alkyl), --NH(OH), --SH,
--SC.sub.1-6 alkyl, --SS(C.sub.1-6 alkyl), --C(.dbd.O)(C.sub.1-6
alkyl), --CO.sub.2H, --CO.sub.2(C.sub.1-6 alkyl),
--OC(.dbd.O)(C.sub.1-6 alkyl), --OCO.sub.2(C.sub.1-6 alkyl),
--C(.dbd.O)NH.sub.2, --C(.dbd.O)N(C.sub.1-6 alkyl).sub.2,
--OC(.dbd.O)NH(C.sub.1-6 alkyl), --NHC(.dbd.O)(C.sub.1-6 alkyl),
--N(C.sub.1-6 alkyl)C(.dbd.O)(C.sub.1-6 alkyl),
--NHCO.sub.2(C.sub.1-6 alkyl), --NHC(.dbd.O)N(C.sub.1-6
alkyl).sub.2, --NHC(.dbd.O)NH(C.sub.1-6 alkyl),
--NHC(.dbd.O)NH.sub.2, --C(.dbd.NH)O(C.sub.1-6 alkyl),
--OC(.dbd.NH)(C.sub.1-6 alkyl), --OC(.dbd.NH)OC.sub.1-6 alkyl,
--C(.dbd.NH)N(C.sub.1-6 alkyl).sub.2, --C(.dbd.NH)NH(C.sub.1-6
alkyl), --C(.dbd.NH)NH.sub.2, --OC(.dbd.NH)N(C.sub.1-6
alkyl).sub.2, --OC(NH)NH(C.sub.1-6 alkyl), --OC(NH)NH.sub.2,
--NHC(NH)N(C.sub.1-6 alkyl).sub.2, --NHC(.dbd.NH)NH.sub.2,
--NHSO.sub.2(C.sub.1-6 alkyl), --SO.sub.2N(C.sub.1-6 alkyl).sub.2,
--SO.sub.2NH(C.sub.1-6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2C.sub.1-6 alkyl, --SO.sub.2OC.sub.1-6 alkyl,
--OSO.sub.2C.sub.1-6 alkyl, --SOC.sub.1-6 alkyl, --Si(C.sub.1-6
alkyl).sub.3, --OSi(C.sub.1-6 alkyl).sub.3-C(.dbd.S)N(C.sub.1-6
alkyl).sub.2, C(.dbd.S)NH(C.sub.1-6 alkyl), C(.dbd.S)NH.sub.2,
--C(.dbd.O)S(C.sub.1-6 alkyl), --C(.dbd.S)SC.sub.1-6 alkyl,
--SC(.dbd.S)SC.sub.1-6 alkyl, --P(.dbd.O).sub.2(C.sub.1-6 alkyl),
--P(.dbd.O)(C.sub.1-6 alkyl).sub.2, --OP(.dbd.O)(C.sub.1-6
alkyl).sub.2, --OP(.dbd.O)(OC.sub.1-6 alkyl).sub.2, C.sub.1-6
alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6
alkynyl, C.sub.3-10 carbocyclyl, C.sub.6-10 aryl, 3-10 membered
heterocyclyl, 5-10 membered heteroaryl; or two geminal R.sup.gg
substituents can be joined to form .dbd.O or .dbd.S; wherein
X.sup.- is a counterion.
[0070] A "counterion" or "anionic counterion" is a negatively
charged group associated with a cationic quaternary amino group in
order to maintain electronic neutrality. Exemplary counterions
include halide ions (e.g., F.sup.-, Cl.sup.-, Br.sup.-, I.sup.-),
NO.sub.3.sup.-, ClO.sub.4.sup.-, OH.sup.-, H.sub.2PO.sub.4.sup.-,
HSO.sub.4.sup.-, sulfonate ions (e.g., methansulfonate,
trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate,
10-camphor sulfonate, naphthalene-2-sulfonate,
naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonic
acid-2-sulfonate, and the like), and carboxylate ions (e.g.,
acetate, ethanoate, propanoate, benzoate, glycerate, lactate,
tartrate, and glycolate).
[0071] Nitrogen atoms can be substituted or unsubstituted as
valency permits, and include primary, secondary, tertiary, and
quarternary nitrogen atoms. Exemplary nitrogen atom substituents
include, but are not limited to, hydrogen, --OH, --OR.sup.aa,
--N(R.sup.cc).sub.2, --CN, --C(.dbd.O)R.sup.aa,
--C(.dbd.O)N(R.sup.cc).sub.2, --CO.sub.2R.sup.aa,
--SO.sub.2R.sup.aa, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.cc)OR.sup.aa, --C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2,
--SO.sub.2N(R.sup.cc).sub.2, --SO.sub.2R.sup.cc,
--SO.sub.2OR.sup.cc, --SOR.sup.aa, --C(.dbd.S)N(R.sup.cc).sub.2,
--C(.dbd.O)SR.sup.cc, --C(.dbd.S)SR.sup.cc,
--P(.dbd.O).sub.2R.sup.aa, --P(.dbd.O)(R.sup.aa).sub.2,
--P(.dbd.O).sub.2N(R.sup.cc).sub.2, --P(.dbd.O)(NR.sup.cc).sub.2,
C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl, or two
R.sup.cc groups attached to a nitrogen atom are joined to form a
3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,
wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 R.sup.dd groups, and wherein R.sup.aa, R.sup.bb, R.sup.cc,
and R.sup.dd are as defined herein.
[0072] In certain embodiments, the substituent present on a
nitrogen atom is a nitrogen protecting group (also referred to as
an amino protecting group). Nitrogen protecting groups include, but
are not limited to, --OH, --OR.sup.aa, --N(R.sup.cc).sub.2,
--C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.cc)R.sup.aa, --C(.dbd.NR.sup.cc)OR.sup.aa,
--C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2, --SO.sub.2N(R.sup.cc).sub.2,
--SO.sub.2R.sup.cc, --SO.sub.2OR.sup.cc, --SOR.sup.aa,
--C(.dbd.S)N(R.sup.cc).sub.2, --C(.dbd.O)SR.sup.cc,
--C(.dbd.S)SR.sup.cc, C.sub.1-10 alkyl (e.g., aralkyl,
heteroaralkyl), C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, and 5-14
membered heteroaryl groups, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aralkyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups,
and wherein R.sup.aa, R.sup.bb, R.sup.cc and R.sup.dd are as
defined herein. Nitrogen protecting groups are well known in the
art and include those described in detail in Protecting Groups in
Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3.sup.rd
edition, John Wiley & Sons, 1999, incorporated herein by
reference.
[0073] For example, nitrogen protecting groups include, but are not
limited to, formamide, acetamide, chloroacetamide,
trichloroacetamide, trifluoroacetamide, phenylacetamide,
3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide,
N-benzoylphenylalanyl derivative, benzamide, p-phenylbenzamide,
o-nitophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide,
(N'-dithiobenzyloxyacylamino)acetamide,
3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide,
2-methyl-2-(o-nitrophenoxy)propanamide,
2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide,
3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine
derivative, o-nitrobenzamide, and
o-(benzoyloxymethyl)benzamide.
[0074] Nitrogen protecting groups also include, but are not limited
to, methyl carbamate, ethyl carbamante, 9-fluorenylmethyl carbamate
(Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate,
9-(2,7-dibromo)fluoroenylmethyl carbamate,
2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl
carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc),
2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl
carbamate (Teoc), 2-phenylethyl carbamate (hZ),
1-(1-adamantyl)-1-methylethyl carbamate (Adpoc),
1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl
carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate
(TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc),
1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2'-
and 4'-pyridyl)ethyl carbamate (Pyoc),
2-(N,N-dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate
(BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl
carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl
carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl
carbamate, N-hydroxypiperidinyl carbamate, alkyldithio carbamate,
benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz),
p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl
carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl
carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl
carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl
carbamate, 2-(p-toluenesulfonyl)ethyl carbamate,
[2-(1,3-dithianyl)]methyl carbamate (Dmoc), 4-methylthiophenyl
carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc),
2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl
carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate,
m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl
carbamate, 5-benzisoxazolylmethyl carbamate,
2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc),
m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate,
o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate,
phenyl(o-nitrophenyl)methyl carbamate, t-amyl carbamate, S-benzyl
thiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate,
cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl
carbamate, p-decyloxybenzyl carbamate, 2,2-dimethoxyacylvinyl
carbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate,
1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate,
1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate,
2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl
carbamate, isobutyl carbamate, isonicotinyl carbamate,
p-(p'-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl
carbamate, 1-methylcyclohexyl carbamate,
1-methyl-1-cyclopropylmethyl carbamate,
1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate,
1-methyl-1-(p-phenylazophenyl)ethyl carbamate,
1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl
carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate,
2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl
carbamate, and 2,4,6-trimethylbenzyl carbamate.
[0075] Nitrogen protecting groups further include, but are not
limited to, p-toluenesulfonamide (Ts), benzenesulfonamide,
2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr),
2,4,6-trimethoxybenzenesulfonamide (Mtb),
2,6-dimethyl-4-methoxybenzenesulfonamide (Pme),
2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte),
4-methoxybenzenesulfonamide (Mbs),
2,4,6-trimethylbenzenesulfonamide (Mts),
2,6-dimethoxy-4-methylbenzenesulfonamide (iMds),
2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc),
methanesulfonamide (Ms), .beta.-trimethylsilylethanesulfonamide
(SES), 9-anthracenesulfonamide,
4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),
benzylsulfonamide, trifluoromethylsulfonamide, and
phenacylsulfonamide.
[0076] Other nitrogen protecting groups include, but are not
limited to, phenothiazinyl-(10)-acyl derivative,
N'-p-toluenesulfonylaminoacyl derivative, N'-phenylaminothioacyl
derivative, N-benzoylphenylalanyl derivative, N-acetylmethionine
derivative, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide,
N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide,
N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane
adduct (STABASE), 5-substituted
1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted
1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted
3,5-dinitro-4-pyridone, N-methylamine, N-allylamine,
N-[2-(trimethylsilyl)ethoxy]methylamine (SEM),
N-3-acetoxypropylamine,
N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary
ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine,
N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr),
N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr),
N-9-phenylfluorenylamine (PhF),
N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino
(Fcm), N-2-picolylamino N'-oxide, N-1,1-dimethylthiomethyleneamine,
N-benzylideneamine, N-p-methoxybenzylideneamine,
N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine,
N--(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine,
N-p-nitrobenzylideneamine, N-salicylideneamine,
N-5-chlorosalicylideneamine,
N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine,
N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine,
N-borane derivative, N-diphenylborinic acid derivative,
N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper
chelate, N-zinc chelate, N-nitroamine, N-nitrosoamine, amine
N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide
(Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates,
dibenzyl phosphoramidate, diphenyl phosphoramidate,
benzenesulfenamide, o-nitrobenzenesulfenamide (Nps),
2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide,
2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide,
and 3-nitropyridinesulfenamide (Npys).
[0077] Exemplary oxygen atom substituents include, but are not
limited to, --R.sup.aa, --C(.dbd.O)SR.sup.aa, --C(.dbd.O)R.sup.aa,
--CO.sub.2R.sup.aa, --C(.dbd.O)N(R.sup.bb).sub.2,
--C(.dbd.NR.sup.bb)R.sup.aa, --C(.dbd.NR.sup.bb)OR.sup.aa,
--C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2, --S(.dbd.O)R.sup.aa,
--SO.sub.2R.sup.aa, --Si(R.sup.aa).sub.3, --P(R.sup.cc).sub.2,
--P(R.sup.cc).sub.3, --P(.dbd.O).sub.2R.sup.aa,
--P(.dbd.O)(R.sup.aa).sub.2, --P(.dbd.O)(OR.sup.cc).sub.2,
--P(.dbd.O).sub.2N(R.sup.bb).sub.2, and
--P(.dbd.O)(NR.sup.bb).sub.2, wherein R.sup.aa, R.sup.bb, and
R.sup.cc are as defined herein. In certain embodiments, the oxygen
atom substituent present on an oxygen atom is an oxygen protecting
group (also referred to as a hydroxyl protecting group). Oxygen
protecting groups are well known in the art and include those
described in detail in Protecting Groups in Organic Synthesis, T.
W. Greene and P. G. M. Wuts, 3.sup.rd edition, John Wiley &
Sons, 1999, incorporated herein by reference. Exemplary oxygen
protecting groups include, but are not limited to, methyl,
t-butyloxycarbonyl (BOC or Boc), methoxylmethyl (MOM),
methylthiomethyl (MTM), t-butylthiomethyl,
(phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM),
p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM),
guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM),
siloxymethyl, 2-methoxyethoxymethyl (MEM),
2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl,
2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP),
3-bromotetrahydropyranyl, tetrahydrothiopyranyl,
1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP),
4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl
S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl
(CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl,
2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl,
1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl,
1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,
2,2,2-trichloroethyl, 2-trimethylsilylethyl,
2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl,
p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl,
3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl,
2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl,
4-picolyl, 3-methyl-2-picolyl N-oxido, diphenylmethyl,
p,p'-dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl,
.alpha.-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl,
di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl,
4-(4'-bromophenacyloxyphenyl)diphenylmethyl,
4,4',4''-tris(4,5-dichlorophthalimidophenyl)methyl,
4,4',4''-tris(levulinoyloxyphenyl)methyl,
4,4',4''-tris(benzoyloxyphenyl)methyl,
3-(imidazol-1-yl)bis(4',4''-dimethoxyphenyl)methyl,
1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl,
9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,
1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxido,
trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl
(TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl
(DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS),
t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl,
triphenylsilyl, diphenylmethylsilyl (DPMS),
t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate,
acetate, chloroacetate, dichloroacetate, trichloroacetate,
trifluoroacetate, methoxyacetate, triphenylmethoxyacetate,
phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate,
4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate
(levulinoyldithioacetal), pivaloate, adamantoate, crotonate,
4-methoxycrotonate, benzoate, p-phenylbenzoate,
2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate,
9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl
2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl
carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec),
2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutyl
carbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkyl
p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl
p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate,
alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl
S-benzyl thiocarbonate, 4-ethoxy-1-napththyl carbonate, methyl
dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate,
4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate,
2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl,
4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate,
2,6-dichloro-4-methylphenoxyacetate,
2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,
2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,
isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate,
o-(methoxyacyl)benzoate, .alpha.-naphthoate, nitrate, alkyl
N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate,
borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate,
sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate
(Ts).
[0078] Exemplary sulfur atom substituents include, but are not
limited to, --R.sup.aa, --C(.dbd.O)SR.sup.aa, --C(.dbd.O)R.sup.aa,
--CO.sub.2R.sup.aa, --C(.dbd.O)N(R.sup.bb).sub.2,
--C(.dbd.NR.sup.bb)R.sup.aa, --C(.dbd.NR.sup.bb)OR.sup.aa,
--C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2, --S(.dbd.O)R.sup.aa,
--SO.sub.2R.sup.aa, --Si(R.sup.aa).sub.3, --P(R.sup.cc).sub.2,
--P(R.sup.cc).sub.3, --P(.dbd.O).sub.2R.sup.aa,
--P(.dbd.O)(R.sup.aa).sub.2, --P(.dbd.O)(OR.sup.cc).sub.2,
--P(.dbd.O).sub.2N(R.sup.bb).sub.2, and
--P(.dbd.O)(NR.sup.bb).sub.2, wherein R.sup.aa, R.sup.bb, and
R.sup.cc are as defined herein. In certain embodiments, the sulfur
atom substituent present on a sulfur atom is a sulfur protecting
group (also referred to as a thiol protecting group). Sulfur
protecting groups are well known in the art and include those
described in detail in Protecting Groups in Organic Synthesis, T.
W. Greene and P. G. M. Wuts, 3.sup.rd edition, John Wiley &
Sons, 1999, incorporated herein by reference.
[0079] In certain embodiments, G.sub.A can be hydrogen, .dbd.O,
.dbd.S, --OR'', --SR'', --NR''H, alkenyl, alkynyl, an amide group,
an ester group, an aldehyde group, a nitrile group, an imino group,
a ketone group, a thione group, an isonitrile group, an
isothiocyanide group, a carbamate group, a thiocarbamate group, or
a cyclic or acyclic, substituted or unsubstituted, branched or
unbranched, (hetero)aliphatic group having 1 to 6 carbon atoms,
wherein each instance of R'' can be independently hydrogen, a
cyclic or acyclic, saturated or unsaturated, substituted or
unsubstituted, branched or unbranched, (hetero)aliphatic group
having 1 to 16 carbon atoms;
[0080] R.sub.A1, R.sub.A2, R.sub.A3, R.sub.A4, R.sub.A5, R.sub.A6,
R.sub.A7, R.sub.A8, R.sub.A9, and R.sub.A10 can each independently
be hydrogen, halogen, or a cyclic or acyclic, substituted or
unsubstituted, branched or unbranched, (hetero)aliphatic group
having 1 to 6 carbons;
[0081] R.sub.A11, R.sub.A12, R.sub.A13, R.sub.A15, R.sub.A16, and
R.sub.A17 can each independently be hydrogen, halogen, or a cyclic
or acyclic, substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 6 carbon atoms;
[0082] R.sub.A14 can be halogen, --NR''H, --SR'', --OR'', alkenyl,
alkynyl, an amide group, an ester group, an aldehyde group, a
nitrile group, an imino group, a ketone group, a thione group, an
isonitrile group, an isothiocyanide group, a carbamate group, or a
thiocarbamate group;
[0083] R.sub.A20 can be hydrogen, halogen, or a cyclic or acyclic,
substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 6 carbon atoms; and
[0084] R.sub.A21 can be hydrogen, halogen, --NR''H, --SR'', --OR'',
alkenyl, alkynyl, an amide group, an ester group, an aldehyde
group, a nitrile group, an imino group, a ketone group, a thione
group, an isonitrile group, an isothiocyanide group, a carbamate
group, a thiocarbamate group, or a cyclic or acyclic, substituted
or unsubstituted, branched or unbranched, (hetero)aliphatic group
having 1 to 6 carbon atoms.
[0085] In certain embodiments, a compound of Formula (I) can be of
Formula (I-A):
##STR00003##
[0086] In certain embodiments, a compound of Formula (I) can be of
the formula:
##STR00004##
[0087] In certain embodiments, a compound of Formula (I) can be of
Formula (I-B):
##STR00005##
[0088] In certain embodiments, a compound of Formula (I) can be of
Formula (I-C):
##STR00006##
[0089] In certain embodiments, a compound of Formula (I) can be of
Formula (I-D):
##STR00007##
[0090] In certain embodiments, G.sub.A can be hydrogen. In certain
embodiments, G.sub.A can be .dbd.O, .dbd.S, --SR'', --OR'',
--N(R'').sub.2, --OH, --SH, or --NH.sub.2. In certain embodiments,
G.sub.A can be can be .dbd.O. In certain embodiments, G.sub.A can
be --OR'' (e.g., --OH, --O(substituted or unsubstituted C.sub.1-6
alkyl), or --OC(.dbd.O)(substituted or unsubstituted C.sub.1-6
alkyl)). In certain embodiments, G.sub.A can be can be .dbd.S. In
certain embodiments, G.sub.A can be --SR'' (e.g., --SH). In certain
embodiments, G.sub.A can be --N(R'').sub.2, --NHR'' (such as
--NH(substituted or unsubstituted C.sub.1-6 alkyl) or
--NHC(.dbd.O)(substituted or unsubstituted C.sub.1-6 alkyl)), or
--NH.sub.2. In certain embodiments, G.sub.A can be can be alkenyl
(e.g., acyclic, substituted or unsubstituted, C.sub.1-6 alkenyl,
such as .dbd.CHC(.dbd.O)O(substituted or unsubstituted C.sub.1-6
alkyl)). In certain embodiments, G.sub.A can be can be a phosphate
group (e.g., --OP(.dbd.O)(acyclic, substituted or unsubstituted,
C.sub.1-6 alkyl).sub.2). In certain embodiments, R.sub.A1 can be
hydrogen. In certain embodiments, R.sub.A1 can be halogen. In
certain embodiments, R.sub.A1 can be an acyclic, substituted or
unsubstituted, branched or unbranched, aliphatic group having 1 to
6 carbons (e.g., acyclic, substituted or unsubstituted, branched or
unbranched, C.sub.1-6 alkyl (such as --CF.sub.3)). In certain
embodiments, R.sub.A1 can be --OR'' (e.g., --OH). In certain
embodiments, R.sub.A1 can be --N(R'').sub.2 (e.g., --NMe.sub.2). In
certain embodiments, R.sub.A2 can be hydrogen. In certain
embodiments, R.sub.A1 and R.sub.A2 can be joined to form .dbd.O. In
certain embodiments, R.sub.A3 can be hydrogen. In certain
embodiments, R.sub.A3 can be acyclic, substituted or unsubstituted,
branched or unbranched, C.sub.1-6 alkyl (e.g., methyl, --CF.sub.3,
--CH.sub.2Br, --CH.sub.2OH, --CH.sub.2OC(.dbd.O)(substituted or
unsubstituted C.sub.1-6 alkyl), or ethyl). In certain embodiments,
R.sub.A3 can be carboxyl. In certain embodiments, R.sub.A4 can be
hydrogen. In certain embodiments, R.sub.A4 can be acyclic,
substituted or unsubstituted, branched or unbranched, C.sub.1-6
alkyl (e.g., methyl, --CF.sub.3, --CH.sub.2C(.dbd.O)(substituted or
unsubstituted C.sub.1-6 alkyl), or ethyl). In certain embodiments,
R.sub.A3 and R.sub.A4 can be joined to form alkenyl (e.g.,
.dbd.CH.sub.2). In certain embodiments, R.sub.A5 can be hydrogen.
In certain embodiments, R.sub.A5 can be acyclic, substituted or
unsubstituted, branched or unbranched, C.sub.1-6 alkyl (e.g.,
methyl or --CH.sub.2OCF.sub.3). In certain embodiments, R.sub.A5
cannot be hydrogen. In certain embodiments, R.sub.A6 can be
acyclic, substituted or unsubstituted, branched or unbranched,
C.sub.1-6 alkyl (e.g., methyl, ethyl, --CH.sub.2OH,
--CH.sub.2C(.dbd.O)Me, or --CH.sub.2C(.dbd.S)Me). In certain
embodiments, R.sub.A6 cannot be --CH.sub.3. In certain embodiments,
R.sub.A7 can be hydrogen. In certain embodiments, R.sub.A7 can be
acyclic, substituted or unsubstituted, branched or unbranched,
C.sub.1-6 alkyl (e.g., methyl). In certain embodiments, R.sub.A7
can be --OR'' (e.g., --OH). In certain embodiments, R.sub.A8 can be
hydrogen. In certain embodiments, R.sub.A8 can be acyclic,
substituted or unsubstituted, branched or unbranched, C.sub.1-6
alkyl (e.g., methyl). In certain embodiments, at least one of
R.sub.A7 and R.sub.A8 cannot be hydrogen. In certain embodiments,
R.sub.A9 can be hydrogen. In certain embodiments, R.sub.A9 can be
acyclic, substituted or unsubstituted, branched or unbranched,
C.sub.1-6 alkyl (e.g., methyl). In certain embodiments, R.sub.A10
can be hydrogen. In certain embodiments, at least one of R.sub.A9
and R.sub.A10 cannot be hydrogen. In certain embodiments, R.sub.A11
can be hydrogen. In certain embodiments, R.sub.A12 can be hydrogen.
In certain embodiments, R.sub.A12 can be an amino group (e.g.,
--N(R'').sub.2, --NHR'', or --NH.sub.2). In certain embodiments,
R.sub.A13 is absent. In certain embodiments, at least one of
R.sub.A12 and R.sub.A13 cannot be hydrogen. In certain embodiments,
R.sub.A14 can be halogen, --NR''H, --SR'', --OR'', alkenyl,
alkynyl, an amide group, a carboxyl group, an ester group, an
aldehyde group, a nitrile group, an imino group, a ketone group, a
thione group, an isonitrile group, an isothiocyanide group, a
carbamate group, or a thiocarbamate group. In certain embodiments,
R.sub.A14 can be hydrogen. In certain embodiments, R.sub.A14 can be
alkyl (e.g., acyclic, substituted or unsubstituted, branched or
unbranched, C.sub.1-6 alkyl (e.g., --CH.sub.2CF.sub.3,
--CH.sub.2OR'' (such as --CH.sub.2OH), --CH.sub.2SR'' (such as
--CH.sub.2SH), --CH.sub.2N(R'').sub.2 (such as --CH.sub.2NHMe or
--CH.sub.2NH.sub.2), --CH.sub.2C(.dbd.O)(acyclic, substituted or
unsubstituted, branched or unbranched, C.sub.1-20 aliphatic (such
as
##STR00008##
or --CH.sub.2C(.dbd.O)N(R'').sub.2 (such as
--CH.sub.2C(.dbd.O)NH(substituted or unsubstituted C.sub.1-6
alkyl))). In certain embodiments, R.sub.A14 cannot be --CH.sub.2OH.
In certain embodiments, R.sub.A14 can be a carboxyl group. In
certain embodiments, R.sub.A14 can be an ester group (e.g.,
--C(.dbd.O)O(acyclic, substituted or unsubstituted, branched or
unbranched, C.sub.1-6 alkyl), such as --C(.dbd.O)OMe). In certain
embodiments, R.sub.A14 can be an aldehyde group. In certain
embodiments, R.sub.A14 can be a ketone group (e.g.,
--C(.dbd.O)-(acyclic, substituted or unsubstituted, branched or
unbranched, C.sub.1-6 alkyl), such as --C(.dbd.O)Me). In certain
embodiments, R.sub.A14 can be a urea group (e.g.,
--NHC(.dbd.O)--NH(substituted or unsubstituted phenyl), such as
--NHC(.dbd.O)--NHPh). In certain embodiments, R.sub.A15 can be
absent. In certain embodiments, R.sub.A14 and R.sub.A15 can be
joined to form .dbd.O or .dbd.S. In certain embodiments, R.sub.A16
can be hydrogen. In certain embodiments, R.sub.A16 can be a
carbamate group (e.g., --NHC(.dbd.O)O(acyclic, substituted or
unsubstituted, branched or unbranched, C.sub.1-6 alkyl), such as
--NHC(.dbd.O)OEt). In certain embodiments, R.sub.A17 can be
hydrogen. In certain embodiments, at least one of R.sub.A16 and
R.sub.A17 cannot be hydrogen. In certain embodiments, R.sub.A20 can
be hydrogen. In certain embodiments, R.sub.A20 can be absent. In
certain embodiments, R.sub.A21 can be hydrogen. In certain
embodiments, R.sub.A21 can be a cyclic or acyclic, substituted or
unsubstituted, branched or unbranched, (hetero)aliphatic group
having 1 to 6 carbon atoms. In certain embodiments, R.sub.A21 can
be acyclic, substituted or unsubstituted, branched or unbranched,
C.sub.1-6 alkyl (e.g., --CH.sub.2OR'' (such as --CH.sub.2OH,
--CH.sub.2C(.dbd.O)OMe, or --CH.sub.2C(.dbd.O)(acyclic, substituted
or unsubstituted, branched or unbranched, C.sub.1-20 aliphatic
(such as
##STR00009##
--CH.sub.2SR'' (such as --CH.sub.2SH);
--CH.sub.2SC(.dbd.O)(substituted or unsubstituted phenyl); or
--CH.sub.2N(R'').sub.2 (such as --CH.sub.2NHR'' (e.g.,
--CH.sub.2NHMe) or --CH.sub.2NH.sub.2)). In certain embodiments,
R.sub.A21 can be acyclic, substituted or unsubstituted, branched or
unbranched, C.sub.2-6 alkenyl (e.g., --CH.dbd.CHCH.sub.3 or
--CH.dbd.NBn). In certain embodiments, R.sub.A21 can be an aldehyde
group. In certain embodiments, R.sub.A21 can be a carboxyl group.
In certain embodiments, each instance of R'' can independently be
hydrogen. In certain embodiments, each instance of R'' can
independently be acyclic, substituted or unsubstituted, branched or
unbranched, C.sub.1-6 alkyl (e.g., methyl, ethyl, propyl, or
butyl).
[0091] The tricyclic ring system
##STR00010##
of Formula (I) may include substituents in addition to one or more
of R.sub.A1 to R.sub.A17, R.sub.A20 to R.sub.A21, and G.sub.A, as
valency permits. In certain embodiments, the tricyclic ring system
of Formula (I) can further be substituted at one or two of the
positions marked with "*":
##STR00011##
In certain embodiments, the tricyclic ring system of Formula (I)
can further be substituted with one or more substituents
independently selected from the group consisting of halogen;
substituted and unsubstituted C.sub.1-6 aliphatic (e.g.,
unsubstituted C.sub.1-6 alkyl, such as --CH.sub.3); and --OR''
(e.g., --OH).
[0092] In certain embodiments, at least one of R.sub.A1, R.sub.A2,
R.sub.A5, R.sub.A7, R.sub.A8, R.sub.A9, R.sub.A10, R.sub.A11,
R.sub.A12, R.sub.A13, R.sub.A15, R.sub.A16, and R.sub.A17 cannot be
hydrogen. In certain embodiments, at least one of R.sub.A3,
R.sub.A4, and R.sub.A6 cannot be --CH.sub.3. In certain
embodiments, when R.sub.A21 is --CHO and G.sub.A is --OH or .dbd.O,
each of R.sub.A14 and R.sub.A15 cannot be --CHO.
[0093] In certain embodiments, the compound of Formula (I) cannot
be of the formula:
##STR00012##
wherein:
[0094] G.sub.A is --OR'' or --N(R'').sub.2;
[0095] R.sub.A14 is an amide group, a nitrile group, an ester
group, or substituted methyl; and
[0096] R.sub.A21 is an aldehyde group, --CH.sub.2OR'', or an ester
group.
[0097] In another aspect, the GLP-1 receptor modulators described
herein are compounds of Formula (II):
##STR00013##
and pharmaceutically acceptable salts thereof, wherein:
[0098] G is hydrogen, .dbd.O, .dbd.S, --NR'H, --SR', or --OR',
wherein R' is hydrogen, an ester group, a ketone group, a thione
group, or a cyclic or acyclic, saturated or unsaturated,
substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 16 carbon atoms;
[0099] W is --O--, --S-- or --NR'--;
[0100] X and Y are each independently a single bond or a saturated
or unsaturated, substituted or unsubstituted, branched or
unbranched, (hetero)aliphatic group having 1 to 3 carbon atoms;
[0101] R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6,
R.sub.7, R.sub.8, R.sub.9, R.sub.12 and R.sub.13 are each
independently hydrogen, halogen, or a cyclic or acyclic,
substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 6 carbon atoms, or R.sub.2 and
R.sub.3 may join to form cycloalkyl, heterocycloalkyl, aryl, or
heteroaryl;
[0102] R.sub.10 and R.sub.11 are each independently hydrogen,
halogen, an amino group, an amide group, an ester group, an
aldehyde group, a nitrile, an imino group, a ketone group, a thione
group, an isonitrile group, an isothiocyanide group, a carbamate
group, a thiocarbamate group, or a cyclic or acyclic, saturated or
unsaturated, substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1 to 6 carbon atoms;
[0103] R.sub.14 is hydrogen or a saturated or unsaturated,
substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1-16 carbon atoms;
[0104] R.sub.15 is hydrogen or a saturated or unsaturated,
substituted or unsubstituted, branched or unbranched,
(hetero)aliphatic group having 1-6 carbon atoms; and
[0105] R.sub.21 is
##STR00014##
or an aldehyde group.
[0106] In certain embodiments, X and Y can each be methylene;
R.sub.1 can be methyl; R.sub.2, R.sub.3 and the two carbon atoms
directly bonded therewith can form a 3,3-dimethyl cyclohexane
ring;
[0107] R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8, R.sub.9,
R.sub.10, R.sub.12, and R.sub.13 can each be hydrogen; R.sub.11 can
be an amide group, an acid group, an ester group, an aldehyde
group, an alcohol group, a carbamate group, a thiocarbamate, a
carbonate, a nitrile group, an amino group, or an imino group; and
the bond between the two carbon atoms directly bonded with R.sub.10
and R.sub.11 can be a double bond.
[0108] In certain embodiments, X and Y can each be methylene;
R.sub.1 can be methyl; R.sub.2, R.sub.3 and the two carbon atoms
directly bonded therewith can form a 3,3-dimethyl cyclohexane ring;
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8, R.sub.9, R.sub.10,
R.sub.12, and R.sub.13 can each be hydrogen; R.sub.11 can be an
amide group, an acid group, an ester group, an aldehyde group, an
alcohol group, a carbamate group, a thiocarbamate, a carbonate, a
nitrile group, an amino group, or an imino group; W can be --O--;
R.sub.12, R.sub.13, R.sub.14, and R.sub.15 can each be hydrogen; G
can be .dbd.O; and the bond between the two carbon atoms directly
bonded with R.sub.10 and R.sub.11 can be a double bond.
[0109] In certain embodiments, a compound of Formula (II) can be of
Formula (II-A):
##STR00015##
[0110] In certain embodiments, a compound of Formula (II) can be of
Formula (II-B):
##STR00016##
[0111] In certain embodiments, a compound of Formula (II) can be of
Formula (II-C):
##STR00017##
[0112] In certain embodiments, a compound of Formula (II) can be of
Formula (II-D):
##STR00018##
[0113] In certain embodiments, G can be can be .dbd.O. In certain
embodiments, G can be --OR' (e.g., --OH, --O(substituted or
unsubstituted C.sub.1-6 alkyl), or --OC(.dbd.O)(substituted or
unsubstituted C.sub.1-6 alkyl)). In certain embodiments, G can be
can be .dbd.S. In certain embodiments, G can be --SR' (e.g., --SH
or --S(substituted or unsubstituted C.sub.1-6 alkyl)). In certain
embodiments, G can be --NHR' (such as --NH(substituted or
unsubstituted C.sub.1-6 alkyl) or --NHC(.dbd.O)(substituted or
unsubstituted C.sub.1-6 alkyl)) or --NH.sub.2. In certain
embodiments, W can be --O--. In certain embodiments, W can be
--S--. In certain embodiments, W can be --NR'-- (e.g.,
--N(substituted or unsubstituted C.sub.1-6 alkyl)- or --NH--). In
certain embodiments, X can be methylene. In certain embodiments, X
can be ethanediyl, vinylene bridge, or propanediyl. In certain
embodiments, Y can be methylene. In certain embodiments, Y can be
ethanediyl, vinylene bridge, or propanediyl. In certain
embodiments, R.sub.1 can be substituted or unsubstituted, branched
or unbranched, C.sub.1-6 alkyl (e.g., methyl, --CH.sub.2OH,
--CH.sub.2C(.dbd.O)Me, or --CH.sub.2C(.dbd.S)Me). In certain
embodiments, R.sub.1 cannot be --CH.sub.3. In certain embodiments,
R.sub.2, R.sub.3, and the two carbon atoms directly bonded
therewith form cycloalkyl (e.g., a 3,3-dimethyl cyclohexane
ring
##STR00019##
that is unsubstituted or substituted (e.g., substituted with
--OH)). In certain embodiments, R.sub.2, R.sub.3, and the two
carbon atoms directly bonded therewith cannot form
##STR00020##
that is unsubstituted. In certain embodiments, R.sub.4, R.sub.5,
R.sub.6, R.sub.7, R.sub.8, R.sub.9, R.sub.10, R.sub.12, and
R.sub.13 can each be hydrogen. In certain embodiments, R.sub.4 can
be hydrogen. In certain embodiments, R.sub.4 can be substituted or
unsubstituted, branched or unbranched, C.sub.1-6 alkyl (e.g.,
methyl or --CH.sub.2OCF.sub.3). In certain embodiments, R.sub.4
cannot be hydrogen. In certain embodiments, R.sub.5 can be
hydrogen. In certain embodiments, R.sub.5 can be substituted or
unsubstituted, branched or unbranched, C.sub.1-6 alkyl (e.g.,
methyl). In certain embodiments, R.sub.5 can be --OR' (e.g., --OH).
In certain embodiments, R.sub.6 can be hydrogen. In certain
embodiments, R.sub.6 can be substituted or unsubstituted, branched
or unbranched, C.sub.1-6 alkyl (e.g., methyl). In certain
embodiments, at least one of R.sub.5 and R.sub.6 cannot be
hydrogen. In certain embodiments, R.sub.7 can be hydrogen. In
certain embodiments, R.sub.7 can be substituted or unsubstituted,
branched or unbranched, C.sub.1-6 alkyl (e.g., methyl). In certain
embodiments, R.sub.8 can be hydrogen. In certain embodiments, at
least one of R.sub.7 and R.sub.8 cannot be hydrogen. In certain
embodiments, R.sub.9 can be hydrogen. In certain embodiments,
R.sub.10 can be hydrogen. In certain embodiments, R.sub.10 can be
an amino group (e.g., --N(R').sub.2, --NHR', or --NH.sub.2). In
certain embodiments, R.sub.11 can be hydrogen. In certain
embodiments, R.sub.11 can be an amide group, an ester group, an
aldehyde group, a carbamate group, a thiocarbamate, a nitrile
group, an amino group, an imino group, or substituted or
unsubstituted, branched or unbranched, C.sub.1-6 alkyl. In certain
embodiments, R.sub.11 can be an ester group, an aldehyde group, a
ketone group, or substituted or unsubstituted, branched or
unbranched, C.sub.1-6 alkyl. In certain embodiments, R.sub.11 can
be substituted or unsubstituted, branched or unbranched, C.sub.1-6
alkyl (e.g., --CH.sub.2CF.sub.3, --CH.sub.2OR' (such as
--CH.sub.2OH), --CH.sub.2SR' (such as --CH.sub.2SH),
--CH.sub.2N(R').sub.2 (such as --CH.sub.2NHMe or
--CH.sub.2NH.sub.2), or --CH.sub.2C(.dbd.O)N(R').sub.2 (such as
--CH.sub.2C(.dbd.O)NH(substituted or unsubstituted C.sub.1-6
alkyl))). In certain embodiments, R.sub.11 cannot be --CH.sub.2OH.
In certain embodiments, R.sub.11 can be an ester group (e.g.,
--C(.dbd.O)O(substituted or unsubstituted, branched or unbranched,
C.sub.1-6 alkyl), such as --C(.dbd.O)OMe). In certain embodiments,
R.sub.11 can be an aldehyde group. In certain embodiments, R.sub.11
can be a ketone group (e.g., --C(.dbd.O)-(substituted or
unsubstituted, branched or unbranched, C.sub.1-6 alkyl), such as
--C(.dbd.O)Me). In certain embodiments, R.sub.12 can be hydrogen.
In certain embodiments, R.sub.13 can be hydrogen. In certain
embodiments, R.sub.14 can be hydrogen. In certain embodiments,
R.sub.14 can be substituted or unsubstituted, branched or
unbranched, C.sub.1-6 alkyl (e.g., --C(.dbd.O)OMe or --C(.dbd.O)
(substituted or unsubstituted, C.sub.2-20 alkenyl). In certain
embodiments, R.sub.21 can be
##STR00021##
In certain embodiments, R.sub.21 can be --CH.sub.2OH. In certain
embodiments, R.sub.21 can be an aldehyde group. In certain
embodiments, R.sub.15 can be hydrogen. In certain embodiments,
R.sub.15 can be absent. In certain embodiments, the bond between
the two carbon atoms directly bonded with R.sub.10 and R.sub.11 can
be a double bond. In certain embodiments, the bond between the two
carbon atoms directly bonded with R.sub.10 and R.sub.11 can be a
single bond. In certain embodiments, R' can be hydrogen. In certain
embodiments, R' can be substituted or unsubstituted, branched or
unbranched, C.sub.1-6 alkyl (e.g., methyl, ethyl, propyl, or
butyl).
[0114] In certain embodiments, at least one of R.sub.4, R.sub.5,
R.sub.6, R.sub.7, R.sub.8, R.sub.9, and R.sub.10 cannot be
hydrogen. In certain embodiments, R.sub.1 cannot be --CH.sub.3. In
certain embodiments, when R.sub.21 is --CHO and G is --OH or
.dbd.O, R.sub.11 cannot be --CHO.
[0115] In certain embodiments, the compound of Formula (II) cannot
be of the formula:
##STR00022##
wherein:
[0116] G is --OR' or --NR'H;
[0117] R.sub.11 is an amide group, a nitrile, an ester group, or
substituted methyl; and
[0118] R.sub.21 is an aldehyde group or --CH.sub.2OR.sub.14.
[0119] In certain embodiments, a compound described herein cannot
be of the formula:
##STR00023##
[0120] In certain embodiments, a compound described herein cannot
be of the formula:
##STR00024##
wherein R.sup.x is a phosphorous-containing group.
[0121] Exemplary compounds described herein include, but are not
limited to:
##STR00025## ##STR00026## ##STR00027## ##STR00028## ##STR00029##
##STR00030## ##STR00031## ##STR00032## ##STR00033## ##STR00034##
##STR00035##
and pharmaceutically acceptable salts thereof, wherein
##STR00036##
[0122] In still another aspect, this present disclosure features
pharmaceutical compositions including one or more of the GLP-1
receptor modulators described herein and a pharmaceutically
acceptable carrier.
[0123] In yet another aspect, this present disclosure features
methods for regulating blood glucose level and/or treating diabetes
in a subject. The method comprises administering to a subject in
need thereof an effective amount of a pharmaceutical composition
described herein. In certain embodiments, the subject is a human
(e.g., a human patient having, at risk for, or suspected of having
diabetes, e.g., type I or type II diabetes.
[0124] In still another aspect, this present disclosure features
methods of treating diabetes in a subject, the method including
administering to a subject in need thereof an effective amount
(e.g., a therapeutically effective amount) of a pharmaceutical
composition described herein.
[0125] The methods described above can also include the step of
identifying a subject in need of the treatment, e.g., a human
patient having or at risk for developing abnormal blood glucose
levels or any disorder associated therewith.
[0126] In further another aspect, the present disclosure features
kits comprising a pharmaceutical composition described herein and
optionally, instructions for using the kits.
[0127] Also within the scope of this present disclosure are a
pharmaceutical composition as described herein for use in
regulating blood glucose level and/or treating diabetes in a
subject, and the use of such a pharmaceutical composition for the
manufacture of a medicament for regulating blood glucose level
and/or treating diabetes in a subject.
[0128] Other features or advantages of the present disclosure will
be apparent from the following detailed description of several
embodiments, and also from the appending claims.
BRIEF DESCRIPTION OF DRAWINGS
[0129] FIG. 1 is a chart showing the effect of GLP-1 (7-36) on
endocytosis medicated by GLP-1 receptor. The
GLP-1R/.beta.-arrestin2 GFP double stable expression U2OS cells
were plated in 384-well plastic plates at a density of 3000
cell/well and cultured overnight. Wells were treated with vehicle
or various concentrations of GLP-1(7-36) for 60 min at room
temperature in the presence (.largecircle.) or absence
(.quadrature.) of 1 .mu.M of exendin 9.
[0130] FIG. 2 shows the activity of an ethanol extract of Hedychium
coronarium (HC) in potentiating the GLP-1 activity. (Panel A) Cell
images of GLP-1 receptor endocytosis elicited by 0 or 4 nM of GLP-1
in the presence or absence of 0.06 mg/ml ethanol extract of HC.
(Panel B) Dose response data of GLP-1 from 0.15 nM to 324 nM on
GLP-1 receptor activation in the presence () or absence
(.box-solid.) of 0.0 6 mg/ml of ethanol extract of HC. (Panel C)
Dose response data of ethanol extract of HC from 0.0025 mg/ml to
0.2 mg/ml on GLP-1 receptor activation by 4 nM of GLP-1.
[0131] FIG. 3 includes charts showing normal phase silica gel
chromatograms of HC ethyl acetate partition fraction. 7 g of EtOAc
partition material was subjected to normal phase silica gel
chromatography and resolved into 37 fractions. The activity was
assayed for each fraction (the ability of 0.002 mg/ml partially
purified compounds to potentiate the receptor endocytosis elicited
by 4 nM of GLP-1) and expressed as % of the maximal activity
elicited by 1 .mu.M of GLP-1. .diamond-solid. represents weight of
each fraction. .box-solid. indicates the activity of each fraction.
I is the pooled fraction 5 to fraction 12.
[0132] FIG. 4 shows an exemplary fractionation of I (the pooled
fraction 5 to fraction 12) on a KROMASIL C.sub.18 column. 1.7 gram
of I was injected into a KROMASIL C.sub.18 column (250.times.50 mm,
10 .mu.m) eluted at flow rate of 109 ml/min with a gradient started
with 57% acetonitrile in water and finally with 100% acetonitrile.
Weight of each fraction (.diamond-solid.) and (.box-solid.)
indicates activity of each fraction.
[0133] FIG. 5 shows exemplary results of a dose response analysis
of galanal B on GLP-1 (Panel A) or PTH (Panel B) induced receptor
endocytosis. The GLP-1R/3-arrestin2 GFP (Panel A) or
PTHR/.beta.-arrestin 2 GFP (Panel B) double stable expression U2OS
cells were plated in 384-well plastic plates at a density of 3000
cell/well and cultured overnight. Dose titration of galanal B on
cell stimulated with 4 nM of GLP-1 7-36 (FIG. 5A) or stimulated
with 15 nM of PTH (Panel B). Panel C shows exemplary results of a
dose response data of GLP-1 from 0.15 nM to 324 nM on GLP-1
receptor activation in the presence () or absence (.box-solid.) of
0.001 mg/ml of galanal B.
[0134] FIG. 6 shows that galanal B potentiated GLP-1 elicited GLP-1
receptor endocytosis. Dose response data of GLP-1 from 0.15 nM to
324 nM on GLP-1 receptor activation in the presence () of 0.001
mg/ml of galanal B (Panel A) or in the absence (.box-solid.) of
0.001 mg/ml of galanal B (Panel B).
[0135] FIG. 7 shows that galanal B, compound 1, and compound 2
increased the potency of GLP-1 dependent receptor endocytosis. Dose
response titration of GLP-1 from 0.15 nM to 324 nM on GLP-1
receptor endocytosis in the presence (.largecircle.), (.gradient.),
(.quadrature.) of 0.0001 mg/ml of galanal B, 0.001 mg/ml of
compound 1, 0.001 mg/ml of compound 2, respectively, or GLP-1 alone
(.tangle-solidup.).
[0136] FIG. 8 shows that exendin 9 diminished positive modulation
effect compound 1, compound 2, and galanal B on GLP-1 elicited
GLP-1 receptor endocytosis. (Panel A) Titration of GLP-1 on GLP-1
receptor endocytosis in the presence of 0.0001 mg/ml of galanal B (
), absence of galanal B (.tangle-solidup.), and in the presence of
0.0001 mg/ml of galanal B plus 1.8 mM of Exendin 9 (.box-solid.).
(Panel B) Titration of GLP-1 on GLP-1 receptor endocytosis in the
presence of 0.001 mg/ml of compound 2 ( ), absence of compound 2
(.tangle-solidup.) and in the presence of 0.001 mg/ml of compound 2
plus 1.8 mM of Exendin 9 (.box-solid.). (Panel C) Titration of
GLP-1 on GLP-1 receptor endocytosis in the presence of 0.001 mg/ml
of compound 1 ( ), absence of compound 1 (.tangle-solidup.) and in
the presence of 0.001 mg/ml of compound 1 and 1.8 mM of Exendin 9
(.box-solid.).
[0137] FIG. 9 shows that compound 1 suppressed, and compound 2
potentiated, GLP-1 dependent cAMP production in RINm5F cells (Panel
A) Dose response titration of GLP-1 in the presence of 0.003 mg/ml
of galanal B (.largecircle.), compound 2 (.diamond.), compound 1
(.gradient.), or GLP-1 alone ( ). (Panel B) Dose response of
galanal (.largecircle.) or compound 2 (.DELTA.) on cAMP production
in RINm5F cells stimulated by 3 nM of GLP-1. (Panel C) Dose
response of compound 1 ( ) on cAMP production in RINm5F cells
elicited by 60 nM of GLP-1.
[0138] FIG. 10 shows that potentiation effect of compound 2 on
GLP-1 elicited cAMP generation is blocked by exendin 9 and MDL
12330A. (Panel A) Titration of exendin 9 on cAMP production in
RINm5F cells stimulated by 2 nM of GLP-1 in the presence (.DELTA.)
and absence ( ) of 0.0025 mg/ml of compound 2. (Panel B) Titration
of GLP-1 on cAMP production in RIMm5F cells (.largecircle.) or in
the presence of 0.0025 mg/ml of compound 2 (a) or in the presence
of 0.0025 mg/ml of compound 2 plus 250 mM of MDL 12330A (b).
[0139] FIG. 11 shows the effect of compound 1 and compound 2 on
cAMP production in RINm5F cells elicited by GIP and Glucagon.
(Panel A) Titration of GIP on the production of cAMP from RINm5F
cells (.DELTA.) or in the presence of 0.025 mg/ml of compound 2
(.quadrature.) or compound 1 (.largecircle.). (Panel B) Titration
of glucagon on the production of cAMP from RINm5F cells
(.tangle-solidup.) or in the presence of 0.025 mg/ml of compound 2
(.box-solid.) or compound 1 ( ).
DETAILED DESCRIPTION
[0140] In an attempt to identify orally active and physiologically
more compliant GLP-1 therapeutics for diabetes (e.g., type II
diabetes) and compounds that selectively regulates the GLP-1
pathway, edible plants were screened for activities that modulate
the GLP-1 receptor signaling. In particular, plant extracts were
screened to identify components that positively modulate GLP-1
receptor signaling in a GLP-1 dependent manner. The compounds
identified in the screening process do not act merely in an
on-and-off manner, as those GLP-1 therapies known in the art.
Rather, the positive modulators identified from plants act more
like a dimmer switch, providing control over the intensity of
activation according to the secreting level of endogenous GLP-1 and
allowing the body to retain its physiological control over
initiating receptor activation.
[0141] Accordingly, described herein are novel GLP-1 receptor
modulators (e.g., activators) such as compounds having Formula (I)
or Formula (II) as described herein, or pharmaceutically acceptable
salts thereof, and uses thereof in regulating blood glucose levels
and treating diabetes such as type I or type II diabetes. The GLP-1
receptor modulators described herein activates GLP-1 receptor only
in the presence of GLP-1, which is different from the GLP-1
independent GLP-1 receptor activators known in the art. See, e.g.,
U.S. Pat. No. 8,501,982.
[0142] Preparation of GLP-1 Receptor Modulators
[0143] The compounds described herein can be prepared by methods
well known in the art. In some examples, the compounds can be
isolated from a native source, such as a plant. In other examples,
the compounds are chemically synthesized following routine
synthetic routes, e.g., by Scheme 1 and Scheme 2 shown below.
[0144] The chemicals used in the above-described synthetic routes
may include, for example, solvents, reagents, catalysts, and
protecting group and deprotecting group reagents. The methods
described above may also additionally include steps, either before
or after the steps described specifically herein, to add or remove
suitable protecting groups in order to ultimately allow synthesis
of the compounds described herein. In addition, various synthetic
steps may be performed in an alternate sequence or order to give
the desired compounds. Synthetic chemistry transformations and
protecting group methodologies (protection and deprotection) useful
in synthesizing applicable compounds described herein are known in
the art and include, for example, those described in R. Larock,
Comprehensive Organic Transformations, VCH Publishers (1989); T. W.
Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis,
3rd Ed., John Wiley and Sons (1999); L. Fieser and M. Fieser,
Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and
Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for
Organic Synthesis, John Wiley and Sons (1995) and subsequent
editions thereof.
[0145] A compound thus obtained can be further purified by methods
known in the art (e.g., flash column chromatography, high
performance liquid chromatography, or crystallization). The
bioactivity of any of the compounds described herein can be
verified via in vitro or in vivo assay systems known in the art,
e.g., those described in the Examples below.
[0146] Pharmaceutical Compositions Comprising GLP-1 Receptor
Modulators and Therapeutic Uses Thereof
[0147] Any of the compounds described herein may be useful in
regulating blood glucose levels or treating diabetes in a subject
via, e.g., modulating the GLP-1 receptor signaling pathways.
[0148] A pharmaceutical composition that includes one or more
compound described herein and a pharmaceutically acceptable
carrier. In certain embodiments, a pharmaceutical composition
described herein includes a compound described herein in an amount
sufficient to regulate blood glucose level in a subject. The
carrier in the pharmaceutical composition must be "acceptable" in
the sense that it is compatible with the active ingredient of the
composition, and preferably, capable of stabilizing the active
ingredient and not deleterious to the subject to be treated. For
example, solubilizing agents such as cyclodextrins, which form
specific, more soluble complexes with the compounds described
herein, or one or more solubilizing agents, can be utilized as
pharmaceutical excipients for delivery of the compounds described
herein. Examples of other carriers include colloidal silicon
dioxide, magnesium stearate, cellulose, sodium lauryl sulfate, and
D&C Yellow #10.
[0149] To practice the methods described herein, an effective
amount of a pharmaceutical composition as described herein can be
administered to a subject in need of the treatment via a suitable
route.
[0150] An "effective amount" is that amount of the one or more
GLP-1 receptor modulator that alone, or together with further
doses, produces the desired response, e.g. reduce the blood glucose
levels in the subject. In the case of treating a particular disease
or condition such as Type I or Type II diabetes, characterized by
dysregulated GLP-1 receptor signaling, the desired response is
inhibiting the progression of the disease or condition. This may
involve only slowing the progression of the disease temporarily,
although more preferably, it involves halting the progression of
the disease permanently. This can be monitored by routine methods
or can be monitored according to routine medical practices. The
desired response to treatment of the disease or condition also can
be delaying the onset or even preventing the onset of the disease
or condition.
[0151] Effective amounts will depend, of course, on the particular
condition being treated, the severity of the condition, the
individual patient parameters including age, physical condition,
size, gender and weight, the duration of the treatment, the nature
of concurrent therapy (if any), the specific route of
administration and like factors within the knowledge and expertise
of the health practitioner. These factors are well known to those
of ordinary skill in the art and can be addressed with no more than
routine experimentation. It is generally preferred that a maximum
dose of the individual components or combinations thereof be used,
that is, the highest safe dose according to sound medical judgment.
It will be understood by those of ordinary skill in the art,
however, that a patient may insist upon a lower dose or tolerable
dose for medical reasons, psychological reasons or for virtually
any other reasons.
[0152] The subject to be treated by any of the methods described
herein can be a human patient, e.g., a human patient having, at
risk for, or suspected of having an elevated blood glucose level or
any disease/condition associated therewith, such as Type I or Type
II diabetes, gestational diabetes, obesity, excessive appetite,
insufficient satiety, and a metabolic disorder. Such a human
patient can be identified by routine medical practices.
Alternatively, the subject can be a non-human mammal, e.g., dog,
cat, cow, pig, horse, sheep, or goat.
[0153] The GLP-1 receptor modulator activates GLP-1 receptor only
in the presence of GLP-1. Thus, when a subject's endogenous level
of GLP-1 is too low, any of the moculators described herein can be
co-administered with GLP-1 or a functional variant thereof.
[0154] The terms "treatment," "treat," and "treating" refer to
reversing, alleviating, delaying the onset of, or inhibiting the
progress of diabetes. In some embodiments, treatment may be
administered after one or more signs or symptoms have developed or
have been observed. In other embodiments, treatment may be
administered in the absence of signs or symptoms of diabetes. For
example, treatment may be administered to a susceptible individual
prior to the onset of symptoms (e.g., in light of a history of
symptoms and/or in light of genetic or other susceptibility
factors). Treatment may also be continued after symptoms have
resolved, for example, to delay or prevent recurrence.
[0155] The pharmaceutical composition described herein can be
administered orally, parenterally, by inhalation spray, topically,
rectally, nasally, buccally, vaginally or via an implanted
reservoir. The term "parenteral" as used herein includes
subcutaneous, intracutaneous, intravenous, intramuscular,
intraarticular, intraarterial, intrasynovial, intrasternal,
intrathecal, intralesional and intracranial injection or infusion
techniques.
[0156] A sterile injectable composition, e.g., a sterile injectable
aqueous or oleaginous suspension, can be formulated according to
techniques known in the art using suitable dispersing or wetting
agents (such as TWEEN 80) and suspending agents. The sterile
injectable preparation can also be a sterile injectable solution or
suspension in a non-toxic parenterally acceptable diluent or
solvent, for example, as a solution in 1,3-butanediol. Among the
acceptable vehicles and solvents that can be employed are mannitol,
water, Ringer's solution and isotonic sodium chloride solution. In
addition, sterile, fixed oils are conventionally employed as a
solvent or suspending medium (e.g., synthetic mono- or
di-glycerides). Fatty acids, such as oleic acid and its glyceride
derivatives are useful in the preparation of injectables, as are
natural pharmaceutically-acceptable oils, such as olive oil or
castor oil, especially in their polyoxyethylated versions. These
oil solutions or suspensions can also contain a long-chain alcohol
diluent or dispersant, or carboxymethyl cellulose or similar
dispersing agents. Other commonly used surfactants such as Tweens
or Spans or other similar emulsifying agents or bioavailability
enhancers which are commonly used in the manufacture of
pharmaceutically acceptable solid, liquid, or other dosage forms
can also be used for the purposes of formulation.
[0157] A pharmaceutical composition for oral administration can be
any orally acceptable dosage form including, but not limited to,
capsules, tablets, emulsions and aqueous suspensions, dispersions
and solutions. In the case of tablets for oral use, carriers which
are commonly used include lactose and corn starch. Lubricating
agents, such as magnesium stearate, are also typically added. For
oral administration in a capsule form, useful diluents include
lactose and dried corn starch. When aqueous suspensions or
emulsions are administered orally, the active ingredient can be
suspended or dissolved in an oily phase combined with emulsifying
or suspending agents. If desired, certain sweetening, flavoring, or
coloring agents can be added. A nasal aerosol or inhalation
composition can be prepared according to techniques well-known in
the art of pharmaceutical formulation and can be prepared as
solutions in saline, employing benzyl alcohol or other suitable
preservatives, absorption promoters to enhance bioavailability,
fluorocarbons, and/or other solubilizing or dispersing agents known
in the art. A pharmaceutical composition described herein can also
be administered in the form of suppositories for rectal
administration.
[0158] Also within the scope of the present disclosure are kits
(e.g., pharmaceutical packs) comprising one or more compound or
pharmaceutical compositions described herein. Such a kit can
further comprise a container (e.g., a vial, ampule, bottle,
syringe, and/or dispenser package, or other suitable container) for
placing the compounds/compositions. In some embodiments, a kit
described herein may include a second container comprising a
pharmaceutically acceptable excipient for dilution or suspension of
a compound or pharmaceutical composition described herein. In some
embodiments, the compound or pharmaceutical composition provided in
the first container and the second container are combined to form
one unit dosage form.
[0159] A kit described herein may include instructions for using
the kit (e.g., for administering a compound or pharmaceutical
composition contained therein to a subject). A kit described herein
may also include information as required by a regulatory agency
such as the FDA. In certain embodiments, the information included
in the kit is prescribing information. A kit described herein may
include one or more additional pharmaceutical agents described
herein as a separate composition.
[0160] A compound or pharmaceutical composition described herein
may be administered concurrently with, prior to, or subsequent to
one or more additional pharmaceutical agents, which may be useful
as, e.g., combination therapies. The additional pharmaceutical
agents may be therapeutically active agents or prophylactically
active agents.
EXAMPLES
[0161] Without intent to limit the scope of the present disclosure,
exemplary compounds and methods of using or making such, as well as
their related results according to the embodiments of the present
disclosure are given below. Note that titles or subtitles may be
used in the examples for convenience of a reader, which in no way
should limit the scope of the present disclosure. Moreover, certain
theories are proposed and disclosed herein; however, in no way
they, should limit the scope of the present disclosure so long as
the present disclosure is practiced according to the present
disclosure without regard for any particular theory or scheme of
action.
Example 1
Synthesis of Exemplary GLP-1 Receptor Modulators
(1)
(.+-.)-(4aS,6aR,11aS,11bR)-7-hydroxy-4,4,11b-trimethyltetradecahydro-1-
H-cyclohepta[a]naphthalene-9-carbaldehyde (RJ-002)
##STR00037##
[0163] A solution of
(.+-.)-(4aS,6aR,11aS,11bR)-9-(hydroxymethyl)-4,4,11b-trimethyl
tetradecahydro-1H-cyclohepta[a]naphthalen-7-ol, TEMPO, and TBACl in
DCM and aqueous solution of NaHCO.sub.3 (0.5M) and K.sub.2CO.sub.3
(0.05M) were vigorously stirred at room temperature. NCS was then
added. Stirring was maintained and the reaction monitored by TLC.
The reaction was quenched with sat. NH.sub.4Cl, the organic layer
was separated, and the aqueous layer was extracted with DCM (three
times). The DCM extracts were washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was coated
on silica gel and purified by flash column chromatography to give
(.+-.)-(4aS,6aR,11aS,11bR)-7-hydroxy-4,4,11b-trimethyltetradecahydro-1H-c-
yclohepta[a]naphthalene-9-carbaldehyde (RJ-002) as a white
solid.
(2) (.+-.)-(6aS,11aS,11bR)-methyl
4,4,11b-trimethyl-7-oxo-2,3,4,4a,5,6,6a,7,8,11,11a,11b-dodecahydro-1H-cyc-
lohepta[a]naphthalene-9-carboxylate (RJ-01)
##STR00038##
[0165] A solution of (.+-.)-(6aS,11aS,11bR)-methy
4,4,11b-trimethyl-7-oxo
2,3,4,4a,5,6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9--
carboxylate (RJ-012) (757 mg, 2.38 mmol) and DBU (0.71 mL, 4.75
mmol) in benzene (48.0 mL) was refluxed for 5 h. The volatiles were
removed in vacuo and the residue was purified by flash
chromatography (EtOAc:hexanes, 1:19) to afford
(.+-.)-(6aS,11aS,11bR)-methyl
4,4,11b-trimethyl-7-oxo-2,3,4,4a,5,6,6a,7,8,11,11a,11b-dodecahydro-1H-cyc-
lohepta[a]naphthalene-9-carboxylate (RJ-011) (691 mg, 91%) as a
white solid. Data for RJ-011: Mp 93-94.degree. C.; IR (film) 2944,
2867, 2845, 1711, 1643, 1436, 1388, 1366, 1259, 1199, 1161, 1122,
1088, 1059 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.13-7.10 (m, 1H), 3.74 (s, 3H), 3.51-3.41 (m, 2H), 2.63 (td,
J=12.0 Hz, J=3.6 Hz, 1H), 2.46-2.37 (m, 1H), 2.16-2.12 (m, 1H),
1.82-1.67 (m, 3H), 1.65-1.52 (m, 2H), 1.52-1.36 (m, 3H), 1.36-1.23
(m, 1H), 1.15 (td, J=13.4 Hz, J=4.0 Hz, 1H), 0.95-0.88 (m, 2H),
0.85 (s, 6H), 0.81 (s, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta. 211.0, 166.9, 162.7, 126.8, 54.6, 52.6, 52.2, 52.1, 41.8,
40.2, 38.4, 37.5, 33.3, 33.2, 29.0, 27.1, 21.4, 20.9, 18.6, 14.3;
HRMS (APCI) calcd for C.sub.20H.sub.30O.sub.3 [M+Na].sup.+:
341.2093. found: 341.2092.
(3) (.+-.)-(6aS,11aS,11bR)-methy
4,4,11b-trimethyl-7-oxo2,3,4,4a,5,6,6a,7,8,11,11a,11b-dodecahydro-1H-cycl-
ohepta[a]naphthalene-9-carboxylate (RJ-012)
##STR00039##
[0167] To a solution of (.+-.)-(6aS,11aS,11bR)-methyl
4,4,11b-trimethyl-7-oxo
tetradecahydro-1H-cyclo-hepta-[a]naphthalene-9-carboxylate (RJ-015)
(872 mg, 2.72 mmol) and NEt.sub.3 (1.5 mL, 10.89 mmol) in
CH.sub.2Cl.sub.2 (27.0 mL) was added TMSOTf (1.0 mL, 5.4 mmol) at
0.degree. C., and stirring was continued for 2 h. The reaction
contents were quenched with sat. NaHCO.sub.3(aq) (60 mL) at
0.degree. C., and the aqueous layer was extracted with
CH.sub.2Cl.sub.2 (3.times.30 mL). The organic extracts were
combined, washed with brine (60 mL), dried with Na.sub.2SO.sub.4,
and concentrated by rotary evaporator. To a solution of the dried
residue in THF (27.0 mL) was added PhSeCl (626 mg, 3.27 mmol) at
-78.degree. C. The mixture was stirred at -78.degree. C. for 30 min
before the addition of pyridine (0.44 mL, 5.45 mmol) and 30%
H.sub.2O.sub.2 (0.48 mL, 5.45 mmol). The mixture was allowed to
warm to 0.degree. C. and stirred for 1 h. The reaction was quenched
by sat. NaHCO.sub.3(aq) (60 mL), and the aqueous layer was
extracted with ether (3.times.30 mL). The organic extracts were
combined, washed with brine (60 mL), dried over Na.sub.2SO.sub.4,
and concentrated in vacuo. The residue was purified by flash
chromatography (EtOAc:hexanes, 4:96) to afford
(.+-.)-(6aS,11aS,11bR)-methy
4,4,11b-trimethyl-7-oxo-2,3,4,4a,5,6,6a,7,8,11,11a,11b-dodecahydro-1H-cyc-
lohepta[a]naphthalene-9-carboxylate (RJ-012) (815 mg, 95%) as a
colorless oil. Data for RJ-012: IR (film) 2925, 2866, 1722, 1667,
1436, 1388, 1366, 1228, 1135, 1022 cm.sup.-1; .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 6.85 (s, 1H), 3.77 (s, 3H), 2.71-2.57 (m, 3H),
1.91-1.78 (m, 3H), 1.73-1.63 (m, 1H), 1.63-1.52 (m, 1H), 1.50-1.28
(m, 5H), 1.28-1.21 (m, 1H), 1.14 (td, J=13.0 Hz, J=4.0 Hz, 1H),
0.96-0.86 (m, 2H), 0.86 (s, 3H), 0.84 (s, 3H), 0.81 (s, 3H);
.sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 208.0, 167.9, 143.2,
135.6, 55.0, 54.8, 52.4, 51.2, 42.0, 38.2, 37.7, 33.3, 33.2, 31.4,
30.8, 25.7, 21.5, 21.4, 18.5, 13.9; HRMS (ESI) calcd for
C.sub.20H.sub.30O.sub.3 [M+H].sup.+: 319.2273. found: 319.2270.
(4)
(.+-.)-(6aS,7R,11aS,11bR)-9-(hydroxymethyl)-4,4,11b-trimethyl-2,3,4,4a-
,5,6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalen-7-ol
(RJ-013) &
(.+-.)-(6aS,7S,11aS,11bR)-9-(hydroxymethyl)-4,4,11b-trimethyl-2,3,4,4a,5,-
6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalen-7-ol
(RJ-014)
##STR00040##
[0169] To a solution of (.+-.)-(6aS,1aS,11bR)-methyl
4,4,11b-trimethyl-7-oxo-2,3,4,4a,5,6,6a,7,8,11,11a,11b-dodecahydro-1H-cyc-
lohepta[a]naphthalene-9-carboxylate (RJ-011) (600 mg, 1.88 mmol) in
DCM (19 mL) at -78.degree. C. was added DIBAL solution (1.0 M in
toluene, 7.5 mL, 7.54 mmol), and the stirring was continued for 3
h. The reaction was quenched by 1N HCl.sub.(aq) (40 mL) and allowed
to warm to room temperature. The phases were separated, and the
aqueous layer was extracted with DCM (3.times.20 mL). The organic
extracts were combined, washed with brine (40 mL), dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by flash chromatography (gradient from 1:9.fwdarw.1:4-2:3
EtOAc:hexanes) to afford
(.+-.)-(6aS,7R,11aS,11bR)-9-(hydroxymethyl)-4,4,11b-trimethyl-2,3,4,4a,5,-
6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalen-7-ol
(RJ-013) (296 mg, 54%) and
(.+-.)-(6aS,7S,11aS,11bR)-9-(hydroxymethyl)-4,4,11b-trimethyl-2,3,4,4a,
5,6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalen-7-ol
(RJ-014) (200 mg, 36%). Both RJ-013 and RJ-014 are white solids.
Data for RJ-013: Mp 134-135.degree. C.; IR (film): 3350, 2918,
2865, 2840, 1652, 1452, 1384, 1123, 1088, 1035, 999 cm.sup.-1;
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.88-5.84 (m, 1H), 3.99
(s. 2H), 3.38-3.33 (m, 1H), 2.53 (dd, J=15.0 Hz, J=8.4 Hz, 1H),
2.38-2.30 (m, 1H), 2.20 (dd, J=15.0 Hz, J=8.4 Hz, 1H), 2.09-2.01
(m, 1H), 1.95-1.80 (m, 2H), 1.72-1.60 (m, 4H), 1.60-1.50 (m, 1H),
1.49-1.41 (m, 1H), 1.41-1.34 (m, 1H), 1.34-1.24 (m, 1H), 1.12 (td,
J=13.0 Hz, J=4.0 Hz, 1H), 1.05-0.91 (m, 2H), 0.90-0.77 (m, 2H),
0.84 (s, 3H), 0.82 (s, 3H), 0.81 (s, 3H); .sup.13C NMR (100 MHz,
CDCl.sub.3) .delta. 136.6, 128.8, 75.0, 68.1, 54.8, 53.5, 48.5,
42.0, 38.8, 37.8, 34.9, 33.4, 33.3, 32.7, 26.3, 21.7, 21.6, 18.9,
14.0; HRMS (MALDI) calcd for C.sub.19H.sub.32O.sub.2 [M+Na].sup.+:
315.2300. found: 315.2314. Data for RJ-014: Mp 89-91.degree. C.; IR
(film): 3354, 2921, 2865, 1449, 1386, 1365, 1062, 1045, 1000, 738
cm.sup.-1; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 5.89 (t, J=7.3
Hz, 1H), 3.95-3.89 (m, 2H), 3.71 (d, J=5.7 Hz, 1H), 2.60-2.54 (m,
1H), 2.44 (dd, J=14.5 Hz, J=7.0 Hz, 1H), 2.16 (dd, J=14.0 Hz, J=8.5
Hz, 1H), 1.90-1.83 (m, 1H), 1.80-1.71 (m, 1H), 1.67-1.56 (m, 3H),
1.56-1.46 (m, 2H), 1.46-1.39 (m, 1H), 1.39-1.26 (m, 1H), 1.16-1.08
(m, 1H), 0.95 (t, J=10.8 Hz, 1H), 0.91-0.77 (m, 2H), 0.84 (s, 3H),
0.81 (s, 3H), 0.81 (s, 3H), 0.80-0.77 (m, 1H); .sup.13C NMR (100
MHz, CDCl.sub.3) .delta. 137.5, 130.1, 70.0, 67.8, 54.5, 47.3,
47.3, 42.0, 39.2, 37.5, 35.4, 33.5, 33.4, 32.1, 27.4, 22.0, 21.5,
18.9, 13.9; HRMS (ESI) calcd for C.sub.19H.sub.32O.sub.2
[M+Na].sup.+: 315.2300. found: 315.2294.
(5) (.+-.)-(6aS,11aS,11bR)-methyl
4,4,11b-trimethyl-7-oxotetradecahydro-1H-cyclo-hepta-[a]naphthalene-9-car-
boxylate (RJ-015)
##STR00041##
[0171] To a solution of
(.+-.)-(4aS,4bR,10aS)-4b,8,8-trimethyldodecahydro-phenanthren-1(4bH)-one
(890 mg, 3.59 mmol) in THF (18 mL) was added LHMDS (1.0 M in THF,
5.4 mL, 5.38 mmol) at -78.degree. C. The mixture was allowed to
warm slowly to 0.degree. C. over the course of 2 h. Then, cooled
down to -78.degree. C., and NCCOOMe (0.43 mL, 5.38 mmol) and TMEDA
(0.73 mL, 5.38 mmol) were added dropwise to the reaction mixture.
Let it slowly warmed to room temperature and the stirring was
continued overnight. The reaction contents were quenched with 1N
HCl.sub.(aq) (40 mL) and extracted with DCM (3.times.20 mL). The
organic extracts were combined, washed with brine (30 mL), dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. To a solution of
Et.sub.2Zn (1.0 M in hexane, 5.4 mL, 5.38 mmol) in DCM (30 mL)
under ice-bath was added neat CH.sub.2I.sub.2. The mixture was
stirred at 0.degree. C. for 1 h, and then the aforementioned crude
in DCM (30 mL) was added. After 5 minutes, removed the ice-bath,
and the stirring was continued for 3 h at room temperature. The
reaction contents were quenched with sat. NH.sub.4Cl.sub.(aq) at
0.degree. C. and allowed to warm to room temperature. The phases
were separated, and the aqueous layer was extracted with DCM
(3.times.30 mL). The organic extracts were combined, washed with
sat. NaHCO.sub.3(aq) (80 mL) and brine (80 mL), dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by flash column chromatography (gradient from
1:48.fwdarw.1:24 EtOAc:hexanes) to afford
(.+-.)-(6aS,11aS,11bR)-methyl
4,4,11b-trimethyl-7-oxotetradecahydro-1H-cyclo-hepta-[a]naphthalene-9-car-
boxylate (RJ-015) (918 mg, 80%) as a white solid. Data for RJ-015:
Mp 67-68.degree. C.; IR (film) 2924, 2852, 1739, 1707, 1461, 1365,
1277, 1199, 1174 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 3.67 (s. 3H), 2.85-2.70 (m, 3H), 2.47 (td, J=12.0 Hz, J=4.0
Hz, 1H), 2.18-2.08 (m, 1H), 1.82-1.71 (m, 3H), 1.70-1.63 (m, 1H),
1.60-1.21 (m, 7H), 1.13 (td, J=13.0 Hz, J=4.0 Hz, 1H), 1.04-0.96
(m, 1H), 0.93-0.85 (m, 2H), 0.84 (s, 3H), 0.81 (s, 3H), 0.79 (s,
3H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 213.9, 174.8, 54.9,
53.9, 53.5, 51.8, 42.1, 42.0, 39.5, 38.6, 37.9, 33.4, 33.3, 29.6,
29.6, 23.8, 21.6, 21.0, 18.8, 14.0; HRMS (EI) calcd for
C.sub.20H.sub.32O.sub.3 [M].sup.+: 320.2351. found: 320.2352.
(6)
(.+-.)-(6aS,11aS,11bR)-4,4,11b-trimethyl-7-oxo-2,3,4,4a,5,6,6a,7,8,11,-
11a,11b-dodecahydro-H-cyclohepta[a]naphthalene-9-carbaldehyde
(RJ-017)
##STR00042##
[0173] To a solution of
(.+-.)-(6aS,7S,11aS,11bR)-9-(hydroxymethyl)-4,4,11b-trimethyl-2,3,4,4a,5,-
6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalen-7-ol
(RJ-014) (6.8 mg, 0.023 mmol) in DCM (0.27 mL) at room temperature
was added Dess-Martin periodinane (29.3 mg, 0.069 mmol), and the
stirring was continued for 6 h. The reaction was quenched by 1M
Na.sub.2SO.sub.3(aq) (0.5 mL) and sat. NaHCO.sub.3(aq) (0.5 mL) and
stirred for another 30 minutes. The phases were separated, and the
aqueous layer was extracted with DCM (3.times.4 mL). The organic
extracts were combined, washed with brine (10 mL), dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by flash chromatography (1:19 EtOAc:hexanes) to afford
(.+-.)-(6aS,11aS,11bR)-4,4,11b-trimethyl-7-oxo-2,3,4,4a,5,6,6a,7,8-
,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carbaldehyde
(RJ-017) (4.4 mg, 65%). Data for RJ-017: Mp 87-88.degree. C.; IR
(film): 2927, 2867, 2837, 1707, 1686, 1639, 1461, 1444, 1388, 1366,
1251, 1152, 1139, 1114 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 9.39 (s, 1H), 6.85-6.82 (m, 1H), 3.45-3.35 (m, 2H),
2.72-2.62 (m, 1H), 2.56 (td, J=12.0 Hz, J=4.0 Hz, 1H), 2.24-2.13
(m, 1H), 1.88-1.79 (m, 2H), 1.79-1.72 (m, 1H), 1.71-1.66 (m, 1H),
1.66-1.58 (m, 1H), 1.54-1.30 (m, 4H), 1.18 (td, J=13.4 Hz, J=4.0
Hz, 1H), 1.00-0.90 (m, 2H), 0.88 (s, 6H), 0.84 (s, 3H); .sup.13C
NMR (100 MHz, CDCl.sub.3) .delta. 210.6, 192.3, 153.8, 137.3, 54.7,
53.9, 51.0, 41.8, 38.6, 37.4, 36.4, 33.3, 33.3, 29.8, 28.6, 21.5,
21.0, 18.7, 14.2; HRMS (EI) calcd for C.sub.19H.sub.28O.sub.2
[M].sup.+: 288.2089. found: 288.2084.
(7)
(.+-.)-(7S,11bR)-7-hydroxy-4,4,11b-trimethyl-2,3,4,4a,5,6,6a,7,8,11,11-
a,11b-dodecahydro-H-cyclohepta[a]naphthalene-9-carbaldehyde
(RJ-018)
##STR00043##
[0175] A solution of
(.+-.)-(6aS,7S,11aS,11bR)-9-(hydroxymethyl)-4,4,11b-trimethyl-2,3,4,4a,5,-
6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalen-7-ol
(RJ-014) (58 mg, 0.20 mmol), TEMPO (3.1 mg, 0.02 mmol), and TBACl
(5.5 mg, 0.02 mmol) in DCM (2 mL) and aqueous solution of
NaHCO.sub.3 (0.5M, 1 mL) and K.sub.2CO.sub.3 (0.05M, 1 mL) were
vigorously stirred at room temperature. NCS (53 mg, 0.40 mmol) was
then added. Stirring was maintained and the reaction monitored by
TLC. After 8 h, the reaction was quenched with sat. NH.sub.4Cl (4
mL), the organic layer was separated, and the aqueous layer was
extracted with DCM (3.times.5 mL). The DCM extracts were washed
with brine (15 mL), dried over Na.sub.2SO.sub.4, and concentrated
in vacuo. The residue was coated on silica gel and purified by
flash column chromatography (gradient from 1:19.fwdarw.1:4
EtOAc:hexanes) to give
(.+-.)-(7S,11bR)-7-hydroxy-4,4,11b-trimethyl-2,3,4,4a,5,6,6a,7,8,11,11a,1-
1b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carbaldehyde (RJ-018)
(43 mg, 75%) as a white solid. Data for RJ-018: Mp 148-150.degree.
C.; IR (film) 3470, 2934, 2918, 2859, 2846, 1671, 1645, 1444, 1387,
1109, 1082, 1046, 737 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 9.39 (s, 1H), 6.96-6.92 (m, 1H), 3.82 (d, J=6.8 Hz, 1H),
2.97 (dd, J=15.2 Hz, J=8.0 Hz, 1H), 2.53 (dd, J=14.6 Hz, J=9.0 Hz,
1H), 2.30 (d, J=15.6 Hz, 1H), 2.11-2.00 (m, 1H), 1.90-1.59 (m, 6H),
1.59-1.49 (m, 1H), 1.49-1.34 (m, 3H), 1.34-1.21 (m, 1H), 1.18-1.07
(m, 2H), 0.90 (dd, J=12.9 Hz, J=3.5 Hz, 1H), 0.85 (s, 3H), 0.84 (s,
3H), 0.81 (s, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
193.8, 156.3, 142.3, 69.8, 54.6, 47.2, 46.1, 41.8, 39.1, 37.7,
33.4, 33.4, 30.8, 29.2, 28.8, 21.8, 21.4, 18.8, 14.0; HRMS (ESI)
calcd for C.sub.19H.sub.30O.sub.2 [M+Na].sup.+: 313.2143. found:
313.2151.
(8)
(.+-.)-(7R,11bR)-7-hydroxy-4,4,11b-trimethyl-2,3,4,4a,5,6,6a,7,8,11,11-
a,11b-dodecahydro-H-cyclohepta[a]naphthalene-9-carbaldehyde
(RJ-019)
##STR00044##
[0177] A solution of
(.+-.)-(6aS,7R,11aS,11bR)-9-(hydroxymethyl)-4,4,11b-trimethyl-2,3,4,4a,5,-
6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalen-7-ol
(RJ-013) (30 mg, 0.10 mmol), TEMPO (1.6 mg, 0.01 mmol), and TBACl
(2.8 mg, 0.01 mmol) in DCM (1 mL) and aqueous solution of
NaHCO.sub.3 (0.5M, 0.5 mL) and K.sub.2CO.sub.3 (0.05M, 0.5 mL) were
vigorously stirred at room temperature. NCS (27.5 mg, 0.21 mmol)
was then added. Stirring was maintained and the reaction monitored
by TLC. After 24 h, the reaction was quenched with sat. NH.sub.4Cl
(4 mL), the organic layer was separated, and the aqueous layer was
extracted with DCM (3.times.5 mL). The DCM extracts were washed
with brine (15 mL), dried over Na.sub.2SO.sub.4, and concentrated
in vacuo. The residue was coated on silica gel and purified by
flash column chromatography (gradient from 1:9.fwdarw.1:4
EtOAc:hexanes) to give
(.+-.)-(7R,11bR)-7-hydroxy-4,4,11b-trimethyl-2,3,4,4a,5,6,6a,7,8,11,11a,1-
1b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carbaldehyde (RJ-019)
(19 mg, 64%) as a white solid. Data for RJ-019: Mp 142-144.degree.
C.; IR (film): 3507, 2963, 2917, 2863, 2844, 1678, 1648, 1436,
1384, 1345, 1302, 1212, 1041, 1007 cm.sup.-1; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 9.38 (s, 1H), 6.96-6.93 (m, 1H), 3.49-3.43 (m,
1H), 2.70 (dd, J=16.0 Hz, J=8.5 Hz, 1H), 2.63-2.49 (m, 2H),
2.30-2.20 (m, 1H), 2.07-2.00 (m, 1H), 1.89-1.82 (m, 1H), 1.79-1.70
(m, 1H), 1.70-1.52 (m, 4H), 1.52-1.43 (m, 1H), 1.43-1.29 (m, 2H),
1.14 (td, J=13.5 Hz, J=3.0 Hz, 1H), 1.09-0.93 (m, 2H), 0.87 (s,
3H), 0.85 (s, 3H), 0.82 (s, 3H), 0.82-0.78 (m, 1H); .sup.13C NMR
(125 MHz, CDCl.sub.3) .delta. 193.9, 157.6, 140.5, 74.4, 54.8,
53.1, 48.1, 42.0, 38.8, 38.0, 33.4, 33.4, 32.6, 28.8, 28.1, 21.7,
21.6, 18.9, 14.0; HRMS (ESI) calcd for C.sub.19H.sub.30O.sub.2
[M+Na].sup.+: 313.2143. found: 313.2137.
(9)
(6aR,1aR,11bS)-methyl6a-(hydroxymethyl)-4,4,11b-trimethyl-7-oxo-2,3,4,-
4a,5,6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carboxy-
late (RJ-022)
##STR00045##
[0179] A mixture of Cp.sub.2TiCl.sub.2 (794 mg, 2.20 equiv.) and
Zinc powder (625 mg, 6.60 equiv) in deoxygenated THF (14 mL) was
stirred at room temperature (30 min) until the red solution turned
green. The green Ti(III) solution was slowly added via cannula to
the stirred solution of (E)-methyl
2-(cyanomethyl)-4-((1R,2R,8aS)-5,5,8a-trimethyl
octahydro-1H-spiro[naphthalene-2,2'-oxiran]-1-yl)but-2-enoate (500
mg, 1.45 mmol) in THF (15 mL) and stirred for 12 h. After this, an
excess of saturated NaH.sub.2PO.sub.4 was added, and the mixture
was stirred for 30 min. The mixture was filtered to remove
insoluble titanium salts. The product was extracted into ether
(3.times.30 mL), and the combined organic layers were washed with
saturated NaHCO.sub.3 (20 mL) and brine, dried over
Na.sub.2SO.sub.4, concentrated and the crude product was column
chromatographied (EtOAc-hexanes, 1:9) to afford
(6aR,1aR,11bS)-methyl6a-(hydroxymethyl)-4,4,11b-trimethyl-7-oxo-2,3,4,4a,-
5,6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carboxylat-
e (RJ-022) as colorless needles (302 mg, 60%).
[0180] Characteristic data of RJ-022: [.alpha.].sup.25.sub.D -21.1
(c 0.93, CHCl.sub.3); mp 179-180.degree. C. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.07 (dt, J=6.2, 3.1 Hz, 1H), 4.14-3.98 (m,
2H), 3.93 (ddd, J=13.9, 6.7, 2.8 Hz, 1H), 3.74 (s, 3H), 3.49 (d,
J=13.9 Hz, 1H), 2.80 (dd, J=8.0, 5.8 Hz, 1H), 2.65-2.44 (m, 2H),
2.18 (dd, J=11.8, 2.1 Hz, 1H), 2.02-1.95 (m, 1H), 1.81 (d, J=12.4
Hz, 1H), 1.66 (ddd, J=14.0, 8.7, 3.8 Hz, 2H), 1.53-1.34 (m, 5H),
1.18 (td, J=13.5, 4.1 Hz, 2H), 0.95 (s, 3H), 0.88 (s, 3H), 0.82 (s,
3H). .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 214.6, 167.0,
143.7, 124.3, 63.3, 56.8, 56.5, 52.2, 50.6, 41.6, 39.8, 37.9, 37.7,
33.4, 33.1, 32.4, 26.3, 21.3, 18.4, 18.1, 16.1. IR (film) 3542,
2935, 1708, 1702, 1640, 1440, 1386, 1263, 1165, 1115, 1060, 753
cm.sup.-1. HRMS (FAB+) calcd for C.sub.21H.sub.33O.sub.4
[(M+H).sup.+] 349.2379. found 349.2380.
(12)
(7S,11aS,11bR)-7-hydroxy-4,4,11b-trimethyl-2,3,4,4a,5,6,6a,7,8,11,11a-
,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carboxylic acid
(RJ-026)
##STR00046##
[0182] A solution of sodium chlorite (12 mg, 0.13 mmol) and
NaH.sub.2PO.sub.4 (41 mg, 0.34 mmol) in H.sub.2O (0.2 mL) was added
dropwise to a rapidly stirred solution of
(.+-.)-(7S,11bR)-7-hydroxy-4,4,11b-trimethyl-2,3,4,4a,5,6,6a,7,8,11,11a,1-
1b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carbaldehyde (RJ-018)
(10 mg, 0.034 mmol) and 2-methyl-2-butene (36 .mu.L, 0.34 mmol) in
tert-butyl alcohol (0.34 mL) at room temperature and the stirring
was continued for 30 h. The reaction mixture was made basic with 3N
NaOH.sub.(aq) and the tert-butyl alcohol was removed in vacuo. The
residue was dissolved in water and extracted twice with hexanes.
The water layer was acidified with 3N HCl.sub.(aq) and extracted
twice with ether. The organic layer was washed with water and
brine, dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The
residue was coated on silica gel and purified by flash column
chromatography (1:9 EtOAc:MeOH) to give
(7S,11aS,11bR)-7-hydroxy-4,4,11b-trimethyl-2,3,4,4a,5,6,6a,
7,8,11,111a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carboxylic
acid (RJ-026) (7.8 mg, 74%) as a white solid. Data for RJ-026: Mp
229-231.degree. C.; IR (film): 3448, 2938, 2865, 2837, 1701, 1458,
1440, 1385, 1363, 1121, 1104, 1039, 1016, 969 cm.sup.-1; .sup.1H
NMR (500 MHz, CD.sub.3OD) .delta. 7.16-7.13 (m, 1H), 3.83-3.80 (m,
1H), 2.93-2.85 (m, 1H), 2.48 (d, J=15.5 Hz, 1H), 2.37 (dd, J=15.0
Hz, J=9.0 Hz, 1H), 2.00-1.87 (m, 2H), 1.84-1.76 (m, 1H), 1.76-1.65
(m, 2H), 1.65-1.57 (m, 1H), 1.50-1.44 (m, 1H), 1.44-1.27 (m, 4H),
1.19 (td, J=13.5 Hz, J=4.0 Hz, 1H), 1.11 (t, J=10.8 Hz, 1H),
0.94-0.89 (m, 1H), 0.88 (s, 3H), 0.87 (s, 3H), 0.85 (s, 3H);
.sup.13C NMR (125 MHz, CD.sub.3OD) .delta. 171.7, 145.1, 133.3,
71.3, 56.3, 49.6, 47.0, 43.2, 40.2, 38.8, 34.4, 34.0, 33.2, 31.1,
28.7, 22.7, 22.3, 20.0, 14.5; HRMS (EI) calcd for
C.sub.19H.sub.30O.sub.3 [M].sup.+: 306.2195. found: 313.2196.
(15) (6aR,1
aR,11bS)-methyl6a-formyl-4,4,11b-trimethyl-7-oxo-2,3,4,4a,5,6,6a,7,8,11,1-
1a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carboxylate
(RJ-029)
##STR00047##
[0184] A solution of (6aR,11aR,11bS)-methyl
6a-(hydroxymethyl)-4,4,11b-trimethyl-7-oxo-2,3,4,4a,5,6,6a,7,8,11,11a,11b-
-dodecahydro-1H-cyclohepta[a]naphthalene-9-carboxylate (40 mg, 0.11
mmol) in DCM (1 mL) was treated with Dess Martin periodinane (70
mg, 0.16 mmol), which was added in portions at room temperature.
After being stirred for 1 h, the mixture was diluted with saturated
aqueous Na.sub.2S.sub.2O.sub.3 (1 mL) and NaHCO.sub.3 (1 mL) were
added. The resulting mixture was stirred vigorously for 30 min and
the layers were separated. The aqueous phase was extracted with
CH.sub.2Cl.sub.2 and the combined organic extracts were washed with
brine and concentrated under vacuum. The residue was column
chromatographied (EtOAc-hexanes, 1:11) to obtain
(6aR,11aR,11bS)-methyl6a-formyl-4,4,11b-trimethyl-7-oxo-2,3,4,4a,5-
,6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carboxylate
(RJ-029) as white solid (80%).
[0185] Characteristic data of RJ-029: mp 86-88.degree. C. IR (film)
2948, 1727, 1693, 1645, 1437, 1389, 1366, 1258, 1115, 1064, 733
cm.sup.-1. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.84 (d, J=1.0
Hz, 1H), 7.16 (dt, J=6.3, 3.1 Hz, 1H), 3.92-3.80 (m, 1H), 3.75 (d,
J=6.6 Hz, 3H), 3.54 (d, J=14.2 Hz, 1H), 2.89-2.65 (m, 2H),
2.40-2.26 (m, 2H), 1.84-1.70 (m, 2H), 1.68-1.58 (m, 1H), 1.53-1.39
(m, 4H), 1.18 (td, J=13.4, 4.3 Hz, 1H), 1.01-0.92 (m, 2H), 0.89 (s,
3H), 0.79 (s, 3H), 0.75 (s, 3H). .sup.13C NMR (100 MHz, CDCl.sub.3)
.delta. 206.2, 199.6, 166.6, 142.6, 125.0, 66.5, 55.4, 52.4, 51.4,
41.6, 38.8, 38.2, 37.6, 33.3, 33.2, 31.2, 25.5, 21.3, 18.8, 18.5,
15.2. HRMS (ES-) calcd for C.sub.21H.sub.29O.sub.4 [(M-H).sup.+]
345.2066. found 345.2059.
(18)
(9Z,12Z)-(7-hydroxy-6a-(hydroxymethyl)-4,4,11b-trimethyl-2,3,4,4a,5,6-
,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-yl)methyl
octadeca-9,12,dienoate
(RJ-033)/(9Z,9'Z,12Z,12'Z)-((6aR,7R,11bS)-(7-hydroxy-4,4,11b-trimethyl-2,-
3,4,4a,5,6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-6a,9--
diyl)bis(methylene)bis(octadeca-9,12-dienoate (RJ-034)
##STR00048##
[0187] To the linoleic acid (LA) (10.4 mg, 0.037 mmol), DMAP (5.5
mg, 0.44 mmol) was added at room temperature, to this,
((6aR,7R,11aR,11bS)-7-hydroxy-4,4,11b-trimethyl-2,3,4,4a,5,6,6a,7,8,11,11-
a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-6a,9-diyl)dimethanol
(12 mg, 0.037 mmol) in CH.sub.2Cl.sub.2 (0.5 mL) was added, stirred
and cooled to 0.degree. C. and DCC (9 mg, 0.044 mmol) was directly
added to the above mixture. The reaction mixture was stirred at
room temperature for overnight and then the mixture was filtered,
washed with CH.sub.2Cl.sub.2 (2 mL). The filtrate was successively
washed with aq. HCl, sat. NaHCO.sub.3 solution and then brine. The
CH.sub.2Cl.sub.2 layer was dried over Na.sub.2SO.sub.4,
concentrated and the resulting residue was column chromatographied
to give
(9Z,12Z)-(7-hydroxy-6a-(hydroxymethyl)-4,4,11b-trimethyl-2,3,4,4a,5,6,6a,-
7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-yl)methyl
octadeca-9,12,dienoate (RJ-033) as colorless oil (40%) and
(9Z,9'Z,12Z,12'Z)-((6aR,7R,11bS)-(7-hydroxy-4,4,11b-trimethyl-2,3,4,4a,5,-
6,6a,7,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-6a,9-diyl)bis(-
methylene)bis(octadeca-9,12-dienoate (RJ-034) as colorless oil
(20%).
[0188] Characteristic data of RJ-033: IR (film) 3346, 3009, 2925,
2854, 1737, 1660, 1646, 1463, 1385, 1169, 1055, 967 cm.sup.-1.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.93 (dd, J=8.2, 3.3 Hz,
1H), 5.50-5.24 (m, 4H), 4.44 (s, 2H), 4.21 (d, J=11.4 Hz, 1H), 3.95
(d, J=11.4 Hz, 1H), 3.52 (d, J=8.1 Hz, 1H), 2.77 (t, J=6.5 Hz, 2H),
2.66 (dd, J=16.3, 8.7 Hz, 2H), 2.44 (d, J=16.4 Hz, 1H), 2.32 (t,
J=7.5 Hz, 2H), 2.27 (dt, J=13.2, 3.1 Hz, 1H), 2.16 (dd, J=16.1,
10.2 Hz, 1H), 2.09-1.98 (m, 4H), 1.75 (d, J=12.5 Hz, 1H), 1.66-1.54
(m, 8H), 1.47-1.25 (m, 14H), 1.13 (td, J=13.5, 4.3 Hz, 1H), 1.04
(tdd, J=13.3, 4.1, 1.5 Hz, 1H), 0.89 (t, J=6.9 Hz, 3H), 0.87 (d,
J=3.6 Hz, 3H), 0.85-0.81 (m, 2H), 0.80 (s, 3H), 0.75 (s, 3H).
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 173.8, 132.6, 132.2,
130.2, 130.0, 128.1, 127.9, 79.4, 69.5, 62.7, 56.3, 55.8, 46.4,
41.9, 39.7, 38.4, 34.3, 33.7, 33.5, 33.4, 33.2, 31.5, 29.6, 29.3,
29.1, 27.2, 25.6, 25.0, 22.9, 22.6, 21.3, 18.6, 18.4, 16.2, 14.1.
HRMS (ES+) calcd for C.sub.38H.sub.64O.sub.4Na [(M+Na).sup.+]
607.4702. found 607.4695.
[0189] Characteristic data of RJ-034: IR (film) 3457, 3007, 2924,
2854, 1737, 1659, 1650, 1454, 1385, 1243, 1163, 1087, 1054, 723
cm.sup.-1. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.91 (d, J=4.7
Hz, 1H), 5.48-5.20 (m, 8H), 4.70 (d, J=11.5 Hz, 1H), 4.45 (d,
J=12.8 Hz, 1H), 4.43 (s, 2H), 3.43 (t, J=8.1 Hz, 1H), 2.77 (t,
J=6.5 Hz, 4H), 2.57 (dd, J=16.3, 8.8 Hz, 1H), 2.39 (d, J=16.2 Hz,
1H), 2.33 (dd, J=15.3, 7.7 Hz, 4H), 2.10-2.00 (m, 8H), 1.77 (d,
J=12.1 Hz, 1H), 1.69-1.57 (m, 4H), 1.44 (d, J=13.9 Hz, 2H),
1.40-1.24 (m, 28H), 1.13 (td, J=13.4, 3.9 Hz, 2H), 0.89 (t, J=6.8
Hz, 6H), 0.86 (s, 3H), 0.83 (s, 3H), 0.80 (s, 3H).
(22) (.+-.)-(6aR,11aR,11bS)-methyl
6a-((methoxycarbonyloxy)methyl)-4,4,11b-trimet-hyl-7-oxotetradecahydro-1H-
-cyclohepta[a]naphthalene-9-carboxylate (RJ-038)
##STR00049##
[0191] To a solution of (.+-.)-(4aR,4bS,10aR)-methyl
10a-((methoxycarbonyloxy)methyl)-4b,8,8-trimethyl-1-oxotetradecahydrophen-
anthrene-2-carboxylate (RJ-037) (23 mg, 0.058 mmol) in DCE (0.6 mL)
was added Et.sub.2Zn (1.0 M in hexane, 93 .mu.L, 0.093 mmol) at
0.degree. C. After 10 minutes, CH.sub.2I.sub.2 (8 .mu.L, 0.093
mmol) was added and the mixture was stirred at 0.degree. C. for 2
h. Then the reaction mixture was allowed to warm to room
temperature and stirred for another 14 h. The reaction contents
were quenched with sat. NH.sub.4Cl.sub.(aq) (2 mL) at 0.degree. C.
and allowed to warm to room temperature. The phases were separated,
and the aqueous layer was extracted with ether (3.times.2 mL). The
organic extracts were combined, washed with sat. NaHCO.sub.3(aq)
(10 mL) and brine (10 mL), dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by flash column
chromatography (gradient from 0:1.fwdarw.1:1 EtOAc:DCM) to afford
(.+-.)-(6aR,11aR,11bS)-methyl 6a-((methoxycarbonyl
oxy)methyl)-4,4,11b-trimethyl-7-oxotetradecahydro-1H-cyclohepta[a]naphtha-
lene-9-carboxylate (RJ-038) (16 mg, 61%) as colorless liquid. Data
for RJ-038: IR (film) 2951, 2868, 2843, 1750, 1704, 1441, 1389,
1367, 1264, 1201, 1175, 1158, 1116, 961, 791 cm.sup.-1; .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 4.72 (d, J=11.2 Hz, 1H), 4.55 (d,
J=11.2 Hz, 1H), 3.76 (s, 3H), 3.69 (s, 3H), 2.99 (dd, J=11.6 Hz,
J=6.8 Hz, 1H), 2.87-2.73 (m, 2H), 2.14-2.04 (m, 1H), 1.84-1.72 (m,
3H), 1.72-1.61 (m, 5H), 1.61-1.57 (m, 1H), 1.52-1.42 (m, 1H),
1.42-1.28 (m, 2H), 1.28-1.21 (m, 1H), 1.15 (td, J=13.4 Hz, J=4.0
Hz, 1H), 0.94-0.89 (m, 1H), 0.87 (s, 3H), 0.87 (s, 3H), 0.81 (s,
3H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 211.2, 175.5,
155.6, 68.5, 56.4, 56.2, 54.8, 54.6, 52.1, 41.7, 39.6, 38.7, 38.5,
38.5, 33.4, 33.2, 31.5, 29.4, 21.4, 21.3, 18.6, 18.1, 16.0; HRMS
(MALDI) calcd for C.sub.23H.sub.36O.sub.6 [M+Na].sup.+: 431.2410.
found: 431.2422.
(23) (.+-.)-(6aR,11aR,11bS)-methyl
6a-((methoxycarbonyloxy)methyl)-4,4,11b-trimeth-yl-7-oxo-2,3,4,4a,5,6,6a,-
7,10,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carboxylate
(RJ-039)
##STR00050##
[0193] To a solution of (.+-.)-(6aR,11aR,11bS)-methyl
6a-((methoxycarbonyloxy)methyl)-4,4,11b-trimethyl-7-oxotetradecahydro-1H--
cyclohepta[a]naphthalene-9-carboxylate (RJ-038) (8.0 mg, 0.020
mmol) in THF (0.2 mL) was added LHMDS (0.5M in THF, 59 .mu.L, 0.029
mmol) at -78.degree. C. The mixture was allowed to warm slowly to
-20.degree. C. over the course of 2 h. Then, cooled down to
-78.degree. C., a solution of PhSeCl (5.6 mg, 0.029 mmol) in THF
(0.05 mL) was added at -78.degree. C. After 3 h, the reaction was
quenched by sat. NaHCO.sub.3(aq) (2 mL), and the aqueous layer was
extracted with ether (3.times.2 mL). The organic extracts were
combined, washed with brine (6 mL), dried over Na.sub.2SO.sub.4,
and concentrated in vacuo. To a solution of the residue mentioned
above in THF (0.4 mL) was added H.sub.2O.sub.2(aq) (4 .mu.L, 0.050
mmol) and pyrine (4 .mu.L, 0.050 mmol) at room temperature. After 2
h, the reaction was quenched by sat. NaHCO.sub.3(aq) (2 mL), and
the aqueous layer was extracted with ether (3.times.2 mL). The
organic extracts were combined, washed with brine (6 mL), dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by flash column chromatography (gradient from
1:49.fwdarw.1:9 EtOAc:hexanes) to give
(.+-.)-(6aR,1.1aR,11bS)-methyl
6a-((methoxycarbonyloxy)methyl)-4,4,11b-trimethyl-7-oxo-2,3,4,4a,5,6,6a,7-
,10,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carboxylate
(RJ-039) (5.0 mg, 63%) as colorless oil. Data for 37: IR (film)
2952, 2865, 2845, 1752, 1721, 1693, 1440, 1389, 1363, 1264, 1210,
1134, 965, 948 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
6.94 (d, J=2.0 Hz, 1H), 4.70 (d, J=11.0 Hz, 1H), 4.59 (d, J=11.0
Hz, 1H), 3.78 (s, 3H), 3.75 (s, 3H), 2.88-2.79 (m, 1H), 2.32-2.21
(m, 1H), 1.96-1.85 (m, 1H), 1.85-1.77 (m, 2H), 1.72-1.65 (m, 1H),
1.65-1.42 (m, 4H), 1.42-1.23 (m, 3H), 1.14 (td, J=13.4 Hz, J=4.0
Hz, 1H), 0.97-0.87 (m, 2H), 0.92 (s, 3H), 0.86 (s, 3H), 0.81 (s,
3H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 207.2, 167.5,
155.5, 138.1, 137.0, 69.0, 56.2, 55.5, 54.9, 54.9, 52.5, 41.7,
39.3, 38.9, 33.4, 33.2, 32.4, 29.5, 21.8, 21.4, 18.5, 18.2, 16.2;
HRMS (ESI) calcd for C.sub.23H.sub.34O.sub.6 [M+Na].sup.+:
429.2253. found: 429.2254.
(24) (.+-.)-(6aR,11aR)-methyl
6a-((methoxycarbonyloxy)methyl)-4,4,11b-trimethyl-7-oxo-2,3,4,4a,5,6,6a,7-
,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carboxylate
(RJ-40)
##STR00051##
[0195] A solution of (.+-.)-(6aR,11aR,11bS)-methyl
6a-((methoxycarbonyloxy)methyl)-4,4,11b-trimethyl-7-oxo-2,3,4,4a,5,6,6a,7-
,10,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carboxylate
(RJ-039) (25 mg, 0.062 mmol) and DBU (19 .mu.L, 0.124 mmol) in
benzene (1.24 mL) was refluxed for 3 h. The reaction was cooled
down to room temperature and then quenched by sat.
NH.sub.4Cl.sub.(aq) (2 mL). The phases were separated, and the
aqueous layer was extracted with EtOAc (3.times.2 mL). The combined
organic layers were washed with sat. NaHCO.sub.3(aq) (5 mL) and
brine (5 mL), dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by flash column chromatography
(gradient from 1:49.fwdarw.1:9 EtOAc:hexanes) to give
(.+-.)-(6aR,1.1aR)-methyl
6a-((methoxycarbonyloxy)methyl)-4,4,11b-trimethyl-7-oxo-2,3,4,4a,5,6,6a,7-
,8,11,11a,11b-dodecahydro-1H-cyclohepta[a]naphthalene-9-carboxylate
(RJ-40) (20 mg, 80%) as colorless oil. Data for RJ-40: IR (film)
2950, 2868, 2843, 1751, 1711, 1645, 1440, 1388, 1367, 1260, 1116,
1069, 963, 790 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.13-7.09 (m, 1H), 4.75 (d, J=11.2 Hz, 1H), 4.63 (d, J=11.2 Hz,
1H), 3.81 (dd, J=13.8 Hz, J=2.2 Hz, 1H), 3.74 (s, 3H), 3.73 (s,
3H), 3.58 (d, J=14.0 Hz, 1H), 2.78-2.66 (m, 1H), 2.60-2.50 (m, 1H),
2.08-2.01 (m, 1H), 1.86-1.78 (m, 1H), 1.73-1.54 (m, 3H), 1.54-1.44
(m, 1H), 1.44-1.23 (m, 3H), 1.16 (td, J=13.4 Hz, J=4.0 Hz, 1H),
0.95 (s, 3H), 0.94-0.88 (m, 2H), 0.87 (s, 3H), 0.82 (s, 3H);
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 207.2, 167.0, 155.4,
144.0, 124.4, 69.3, 56.3, 55.8, 54.8, 52.2, 52.1, 41.6, 39.8, 38.3,
37.5, 33.7, 33.4, 33.1, 26.6, 21.3, 18.5, 18.5, 15.6; HRMS (EI)
calcd for C.sub.23H.sub.34O.sub.6 [M+Na].sup.+: 429.2253. found:
429.2260.
Example 2
Characterization of Bioactivities of Exemplary GLP-1 Receptor
Modulators
[0196] Receptor endocytosis following arrestin recruitment and cAMP
production subsequent to G.alpha.s coupling are two major immediate
downstream cellular pathways upon GLP-1 receptor activation.
Arrestin recruitment will lead to proliferation and anti-apoptosis
of pancreatic b-cells (30, 31), while production of cAMP will lead
to insulin secretion (Doyle et al., Pharmacology &
Therapeutics, 2007, 113, 546). In this example, the
.beta.-arrestin2-GFP biosensor technology (45) was employed in
screening for a plant extract library to identify those that can
potentiate GLP-1 to elicit receptor endocytosis. Upon agonist
binding to GPCR, the cytoplasmic arrestin rapidly translocate to
and bind to the activated GPCR. Arrestin also mediates receptor
internalization by targeting the receptor to clathrin-coated pits
(46) which is a convergent step of GPCR activation.
.beta.-arrestin2-GFP biosensor technology involved less steps of
enzymatic cascade and yielded more information on the compound
(45), thus was used in the initial screening of plant crude
extracts and also used as an assay for purifying active compounds
from plant crude extracts there would found to be active in
eliciting receptor endocytosis. Following the activity of this
assay, a Hedychium coronarium (HC) extract was identified to
potentiate GLP-1 in arrestin mediated GLP-1 receptor endocytosis.
Furthermore, compounds were isolated and purified from HC extract
to homogeneity by activity directed fractionation, one of the
active compounds was identified to be galanal B. Abilities of
synthetic galanal B and its analogs to modulate GLP-1 dependent
cAMP production in RINm5F cells and to modulate GLP-1 dependent
receptor endocytosis were compared. This analysis revealed that by
modifying structure of galanal B, novel compounds can be generated
that selectively potentiate or suppress GLP-1 in G.alpha.s coupling
pathway. Since it is well documented that type II diabetes still
retain their ability to secret GLP-1 (43,44), it is expected that
compound positively modulates GLP-1 by increasing the potency of
GLP-1 should be potential drug of choice in anti-diabetics.
Materials and Methods
[0197] Extraction of HC Leaves
[0198] Dried leaves of HC (2 kg) was minced and extracted with
ethanol (20 L) at room temperature with constant stirring for 2
days. The extract was filtered off and concentrated to give a
residue that was suspended in 500 ml of 80% ethanol and partitioned
with 500 ml of n-hexane for three times. The remaining was
concentrated, suspended in 500 ml of water and partitioned with 500
ml ethyl acetate three times followed by 500 ml n-BuOH three times.
7 g of ethyl acetate fraction was chromatographed on a silica-gel
column (4.5 cm.times.21 cm, 180 g MERCK 200-400 mesh silica gel)
eluted with 1200 ml of 20% and 1200 ml of 30% hexane-EtOAc each,
followed by 1600 ml of 50%, 800 ml of 80% hexane-EtOAc and 800 ml
of 100% EtOAc, the column was further eluted with 800 ml each of
20% and 50% methanol: EtOAc; 200 ml was collected for each
fraction.
[0199] Chemical Synthesis of Galanal B and its Analogues
[0200] Commercially available (.+-.)-sclareolide (3) was selected
as the starting material and readily converted, as shown in Scheme
1, to olefin 5 in 68% yield through a two-step protocol (1, 2) and
subsequently reduced by LiAlH.sub.4 to afford aldehyde 6 in 90%
yield. With considerable optimization, the subjection of 6 into a
solution of ylide 7 in hot toluene efficiently furnished Wittig
adduct 8 as a single isomer in 85% yield. The terminal double bond
of compound 8 was then epoxidized selectively by m-CPBA, giving an
inseparable mixture of 9 and 10 in a ratio of 10:1. When the
mixture of epoxides 9 and 10 was subjected to Cp.sub.2TiCl.sub.2
and Zn metal (3-7), the Ti(III) species generated in situ would
react with oxirane to afford the homolytic cleavage of the more
substituted C--O bond, giving the more stable tertiary radical
intermediate. Subsequently, the ensuing equatorial addition of the
-titanoxyl radical to the nitrile caused the generation of imine
radical, which evolved into the corresponding ketone. Compound 11
was obtained exclusively in 60% yield with trace amount of side
products originated from ring opening of oxirane. The structural
connectivity of 11 was confirmed by single-crystal X-ray
diffraction. DIBAL-H reduction of 11 to compound 2 (a mixture of
triols 12 and 13) followed by selective oxidation of primary
alcohols by TEMPO (8) furnished 1:5 ratio of galanal A and galanal
B, which can be separated by column chromatography and are
identical in all respect with authentic samples isolated from HC
plant and the reported data (9, 10).
##STR00052##
[0201] Compound 2 was synthesized according to the method shown in
Scheme 2.
[0202] MeNHOMe.HCl (2 equiv.), Me.sub.3Al (2 equiv.),
CH.sub.2Cl.sub.2, 0.degree. C. to rt (room temperature), 85%. b)
SOCl.sub.2 (5 equiv.), pyridine (10 equiv.), CH.sub.2Cl.sub.2,
-78.degree. C., 80%. c) LiAlH.sub.4 (2 equiv.), THF, rt, 90%. d) 10
(3 equiv.), toluene, reflux, 85%. e) m-CPBA (2 equiv.),
CH.sub.2Cl.sub.2, rt, 85%. f) Cp.sub.2TiCl.sub.2 (2.2 equiv.), Zn
(6.6 equiv.), THF, rt, 60%. g) DIBAL-H (8 equiv.),
CH.sub.2Cl.sub.2, -78.degree. C., 80%. h) TEMPO (0.2 equiv.), NCS
(4 equiv.), TBACl (0.2 equiv.), NaHCO.sub.3, K.sub.2CO.sub.3,
CH.sub.2Cl.sub.2, rt, 70%).
##STR00053## ##STR00054##
[0203] Receptor Endocytosis Assay
[0204] U2OS osteosarcoma cell line stably expressing a
.beta.-arrestin2:GFP fusion protein was obtained from Norak
Biosciences (now Molecular Devices, part of MDS Inc., Mississauga,
Ontario). GLP-1 receptor expression construct was used to transfect
the U2OS cell stably expressing .beta.-arrestin2:GFP fusion protein
and to obtain cell line stably co-expressing GLP-1 receptor and
.beta.-arrestin2:GFP fusion protein. High-content imaging of
receptor endocytosis in cells was conducted with 0.03 mg/ml of
ethanol extract from 2500 edible plant to identify potentiating
activity for the GLP-1 dependent GLP-1 receptor endocytosis.
Extracts were supplied at a concentration of 100 mg/ml in 100%
DMSO. Three replicate 384-well assay microplates were plated with
U2OS cells stably expressing GFP-.beta.-arrestin2 fusion protein
and the GLP-1 receptor at a density of 3,000 cells per well.
Aliquots of 2.5 .mu.L of 10.times. stocks of plant extract in
phenol red free MEM containing 1, 0.3, or 0.1 mg/ml of plant
extract plus 40 nM of GLP-1 were transferred to each well of the
cell assay plate, which contained 22.5 .mu.L of phenol red free
MEM. The 3 cell assay plates were incubated at room temperature for
60 min before fixation with 2% formaldehyde and labeling of the
cell nuclei with 5 .mu.g/mL of the DNA-binding dye Hoechst 33342
(Molecular Probes, Eugene, Oreg.) for 1 hr. Plates were washed with
PBS twice and sealed and could be stored at 4.degree. C. The final
concentration of the extract in the cell plate was 0.1, 0.03 and
0.001 mg/ml, and the final DMSO concentration was 1%.
[0205] Bioluminescence Resonance Energy Transfer (BRET) Assay
[0206] BRET assays were performed to examine the effect of
candidate GLP-1 receptor modulators on the intracellular cAMP
levels in RINm5F cells (an insulin-secreting cell line), following
routine technology. See, e.g., Bertrand et al., J. Recept Signal
Transduct Res., 2002, 22(1-4):533-541; Barak et al., Mol.
Pharmacol., 74(3):585-594 (2008); U.S. Pat. No. 8,647,887, and
WO1999066324.
[0207] Imaging and Analysis
[0208] Images and data of the cells were performed according to
reported methods (51), using an ArrayScan.RTM. VTI HCS Reader
(Cellomics, Inc. Pittsburgh, Pa.). Appropriate filter sets for
detection of the 2 fluorophores were used, and the different
fluorescent signals were recorded in 2 different image collection
channels of the ARRAYSCAN VTI HCS Reader (i.e., channel 1 contained
the blue fluorescent Hoechst 33342-labeled nuclear images, and
channel 2 contained the green fluorescent GFP-.beta.-arrestin
images). A 20.times.0.4 numerical aperture microscope objective was
used for the imaging, 3 fields were imaged per well, and
CELLOMICS's Spot Detector BIOAPPLICATION was used to acquire and
analyze the images. For these experiments, the Spot Detector
BIOAPPLICATION used the Hoechst-labeled nuclei to identify
individual cells and then automatically counted and analyzed the
GFP-labeled spots associate with each cell. In addition to the
number of spots and the sum of their areas and pixel intensities,
the BIOAPPLICATION also reports properties of the individual nuclei
such as their area. The extent of receptor endocytosis response was
expressed as % of that elicited by 1 .mu.M of GLP-1.
[0209] One unit of activity is defined as the activity that will
reach 50% of maximal response in a well of 384-well plate.
Results
[0210] Primary Screening Herb Ethanol Extracts that are Able to
Potentiate GLP-1 Signaling
[0211] To identify dietary molecules that could potentiate GLP-1
dependent receptor signaling, an ethanol extract library consisting
2500 edible plants was screened using .beta.-arrestin2-GFP
biosensor technology which is based on the observation that the
.beta.-arrestin2 binding of an activated receptor is a convergent
step of GPCR signaling (45, 46). By monitoring the binding of
.beta.-arrestin2-GFP to the activated GLP-1 receptor and the
following .beta.-arrestin-mediated internalization of the activated
receptors to clathrin-coated pits, a dose dependent activation of
GLP-1 receptor by GLP-1 was observed. In addition, these processes
can be visualized from its image (52), thus easily exclude false
positive hits. As shown in FIG. 1, GLP1 (7-37) activates GLP 1
receptor dependent .beta.-arrestin2 translocation in a dose
dependent and saturable manner, and the EC.sub.50 was measured to
be 10 nM of GLP-1. This analysis demonstrated that 4 nM of GLP1
(7-37) is able to activate GLP-1 receptor to the level of 10 to 20%
of the maximal response by 1 .mu.M of the peptide. To screen for
extract that acts as a modulator to potentiate GLP-1 concentration
dependent GLP1 receptor endocytosis, 0.1 mg/ml of plant ethanol
extract was used to test its ability to enhance the agonistic
effect of 4 nM of GLP-1 (7-37) on cells co-expressing GLP-1
receptor and .beta.-arrestin2-GFP. 2500 herb ethanol extracts were
screened for ability to enhance the GLP-1 receptor endocytosis
elicited by 4 nM of GLP-1. 25 out of 2500 herb extracts were found
to enhance the agonistic activity of 4 nM GLP-1 (7-37) from 20% to
more than 80% of the maximal response, however, 9 out these 25
primary hits were false positive as judged by visualizing the
corresponding images. The remaining 16 positive hits were tested
for their selectivity to potentiate GLP-1 receptor by assaying
their effect on PTHR, GIPR and BRS3 signaling. These selectivity
tests revealed that 11 out these 16 positive hits will also
activate PTHR or GIPR or BRS3, thus the remaining 5 plant extracts
were found specifically potentiate GLP-1 receptor signaling. 4 out
of these 5 plants have been documented to display hypoglycemic
effect or anti-diabetic effect on rodent or on other mammalian
species, HC is the only plant has not been reported for its effect
on blood glucose excursion. HC ethanol extract was further
subjected to characterizing its effects on the potency and efficacy
of GLP-1 signaling. As shown in FIG. 2A, GFP-arrestin is evenly
distributed in the cytosol of cells stably co-expresses
.beta.-arrestin1-GFP and GLP-1 receptor when the receptor is at
resting stage, addition of 0.06 mg/ml of HC extract alone do not
change the distribution of .beta.-arrestin1-GFP, stimulation by 4
nM of GLP-1 leads to low level formation of vesicles contains
(3-arrestin1-GFP and GLP-1 receptor in the cytosol and perinuclear
region, much more vesicles of the receptor/.beta.-arrestin1-GFP
complex was observed if cell co-incubated with 4 nM GLP-1 and 0.06
mg/ml of HC extract. FIG. 2B reveals titration of GLP-1 on GLP-1
receptor activation responses as % of that stimulated by 1 .mu.M of
GLP-1, showing that activation increased as GLP-1 increased from
1.5 nM and reached saturation at 324 nM of GLP-1, revealing that
GLP-1 elicits GLP-1 receptor endocytosis in a dose dependent and
saturable manner. While in the presence of 0.06 mg/ml ethanol
extract of HC, the receptor activation started with 0.44 nM of
GLP-1 and reached saturation at a GLP-1 concentration of 10 nM.
Comparison of the dose response data of GLP-1 titration revealed
that EC.sub.50 was reduced from 10.7 nM to 3.8 nM and that maximal
activation increased from 88.2% to 129% by the presence of 0.06
mg/ml of ethanol extract of HC. The effect of HC ethanol extract is
highly dependent on the concentration of GLP-1, since HC plant
extracts alone does not elicit GLP-1 receptor endocytosis,
indicating that it behaves like a potentiator rather than an
agonist on GLP-1 receptor. This analysis indicated HC ethanol
extract potentiate GLP-1 signaling by increasing the efficacy and
potency of GLP-1. Further, the potency of HC extract on GLP-1
signaling was also evaluated. A titration of HC extract was
performed on the receptor endocytosis elicited by 4 nM GLP-1. FIG.
2C revealed the dose-response analysis of the titration of HC
extract on receptor activation by 4 nM GLP-1; 4 nM of GLP-1 alone
led to 20% of receptor activation while as the concentration of HC
ethanol extract increased to 0.022 mg/ml the receptor activation
increased, and when HC increased to 0.2 mg/ml the activation by 4
nM of GLP-1 was potentiated from 20% to 70%. This analysis revealed
that the HC ethanol extract potentiated GLP-1 activity in a dose
dependent and saturable manner with an EC.sub.50 for GLP-1
signaling around 0.038 mg/ml and that maximal potentiation
stimulation up to 70% of that of maximal GLP-1 titration. The
potentiation effect of HC required the presence of GLP-1 as the
ethanol extract of HC alone do not elicit any receptor endocytosis
and has no effect on the distribution of GFP-arrestin in U2OS cells
(FIG. 1A).
[0212] Isolation and Purification of Active Components from HC
Ethanol Extract
[0213] Since extract from HC displayed potent activity on GLP-1
elicited receptor endocytosis, the active components was isolated
according to its effect to potentiate GLP-1 elicited receptor
endocytosis. Solvent partition with hexane, ethyl acetate, butanol,
and water revealed that most of the activity was recovered in the
ethyl acetate fraction (Table 1), while little activity was noted
in the layer of dH.sub.2O. There is a 6-fold increase in the
affinity of the fraction to potentiate GLP-1 elicited GLP-1
receptor endocytosis in ethyl acetate fraction as the EC.sub.50 of
the fraction was reduced from 0.045 mg/ml to 0.007 mg/ml. The
recovery of the activity was more than 100% in this step of
fractionation, indicating some of the negative activity was
removed. Chromatography of the ethyl acetate fraction on silica gel
resolved into 36 fractions, activity assay of each fraction showed
a significant activity was recovered between fraction 4 to fraction
12 (FIG. 3). These active fractions (fraction I) were pooled and
subjected to reverse phase silica gel chromatography and resolved
into 106 fractions (FIG. 4). There are 30 fractions showing
activity significantly higher than that of 4 nM GLP-1 alone.
Fraction 26 is one of the fractions able to potentiate GLP-1
response from 20% to 40% at a concentration of 0.0002 mg/ml and its
EC.sub.50 to potentiate 4 nM of GLP-1 was measured to be
EC.sub.50=0.00024 mg/ml, and its potentiation effect is selective
for GLP-1 but not for PTH (FIGS. 5 A and 5B). Since this fraction
displayed purity more than 90% thus was subjected to structure
elucidation, and its structure turn out to be galanal B which
increase the affinity of GLP-1 by 4 folds (FIG. 5 C).
TABLE-US-00001 TABLE 1 Partition of ethanol extract of HC EC.sub.50
at 4 nM Specific Total Weight GLP1 Activity Activity HC Fraction
(gram) (mg/ml) .sup.a(U/mg) .sup.b(U) .times. 10.sup.3 Dried plant
1400 Ethanol extract 209.65 0.045 0.42 .times. 10.sup.3 8.8 .times.
10.sup.7 Hexane 33.42 0.032 1.25 .times. 10.sup.3 4.2 .times.
10.sup.7 Ethyl acetate 19.3 0.007 5.71 .times. 10.sup.3 1.1 .times.
10.sup.8 Butanol 28.6 nd nd nd dH.sub.2O 124.6 nd nd .sup.aOne
arbitrary unit is defined as the activity that will induce 50% of
maximal response in a well of 384-well plate with a volume of 25
.mu.L for each well. .sup.bTotal activity is obtained by
multiplying the specific activity to the total weight.
[0214] Characterization of Galanal B, Compound 1, and Compound
2
[0215] Galanal B, Compound 1, and Compound 2 were synthesized as
described above. Their effects on GLP-1 induced receptor
endocytosis were investigated. As shown in FIG. 7, dose response
curve of GLP-1 and receptor endocytosis was left shifted by the
presence of 0.003 mg/ml of these compounds. The EC.sub.50 values
and efficacy of galanal B, compound 1, and compound 2 are shown in
Table 2 below. These results show that the potentiating effect of
these compounds on GLP-1 dependent receptor endocytosis is highly
dependent on the presence of GLP-1 since reduced levels of GLP-1
resulted in a decrease of the activity. The potentiating effects of
these compounds are all blocked by GLP-1 receptor
antagonist--exendin 9 (FIG. 8), indicating their effects are
mediated via GLP-1 receptor.
TABLE-US-00002 TABLE 2 Effect of galanal B, compound 1, and
compound 2 on the EC.sub.50 and efficacy of GLP-1 to elicit
receptor endocytosis GLP-1 Compound 2 + Galanal B + Compound 1 +
only GLP-1 GLP-1 GLP-1 EC.sub.50 (nM) 8.1 0.80 1.1 4.8 Efficacy
83.2 82 115.2 106.6 (100%)
[0216] GLP-1 receptor is coupled to G.alpha.s and leads to
generation of cAMP in pancreatic .beta. cells, to examine if the
receptor endocytosis potentiating activities of the present
compounds will translate into the ability to potentiate cAMP
production, the effect of these compounds on the GLP-1 induced
intracellular cAMP generation in RINm5F cell was tested via a BRET
assay, using a cAMP biosensor as known in the art, which monitored
the bioluminescence energy transferring as intracellular cAMP level
increased (53). It has been demonstrated that cAMP binding induced
a remarkable conformational change of the cAMP sensor expressed in
RINm5F cells and the conformational change was determined by
measuring the bioluminescence resonance energy transfer between the
donor and acceptor in the cAMP sensor. In the resting stage when
the intracellular cAMP is minimal, a large BRET ratio was observed.
When cells were incubated with GLP-1 or forskolin, an increase of
the intracellular levels of cAMP and a decrease of the BRET ratio
were observed. However, this dose response was eliminated by the
presence of 250 .mu.M of adenylyl cyclase inhibitor MDL 12330A. To
analyze the effect of compounds on the GLP-1 elicited cAMP
production, it was examined if the potency of GLP-1 be changed by
the presence of galanal b, compound 1, and compound 2. As shown in
FIG. 9A, the dose response curve of GLP-1 was left shifted by 2.5
orders of magnitude when cells were co-incubated with 0.0025 mg/ml
of compound 2 and EC.sub.50 was reduced from 7.8 nM to 0.0025 nM.
The effect of compound 2 on GLP-1 elicited cAMP production
decreased as GLP-1 concentration reduced and was diminished when
GLP-1 concentration reduced to 0.1 pM, indicating that compound 2
alone is not able to induce cAMP production, but to increase the
potency of GLP-1 to stimulate cAMP production. The potentiating
effect of compound 2 on GLP-1 stimulated cAMP production was
blocked by the presence of GLP-1 receptor antagonist--exendin 9
(FIG. 10A) or by MDL12330A--cyclase inhibitor (FIG. 10B),
indicating the requirement of GLP-1 receptor and production of cAMP
in the potentiation by compound 2. To measure the affinity of
compound 2 to potentiate GLP-1 dependent cAMP production, the
effect of compound concentration on the enhancement of cAMP
production elicited by 3 nM of GLP-1 was titrated. As shown in FIG.
9B, the production of cAMP by 3 nM of GLP-1 increased as the
concentration of compound increased and became saturated as the
concentration of compound 2 reach 0.03 mg/ml. The affinity of
compound 2 to enhance GLP-1 was determined to be 0.001 mg/ml.
Similar dose of galanal B did not facilitate GLP-1 to stimulate
cAMP production (FIG. 9A), galanal B did not significantly affect
the dose-response curve of GLP-1 and cAMP production by 3 nM of
GLP-1 was not changed by galanal B up to 0.04 mg/ml (FIG. 9B). By
contrast, compound 1 remarkably suppresses GLP-1 elicited cAMP
production in RINm5F cells and the dose response curve of GLP-1 on
cAMP production was right shifted by almost 2 orders of magnitude,
the EC.sub.50 was increased from 7.8 to 360 nM (FIG. 9A). The
affinity of compound 1 on GLP-1 elicited cAMP production was
obtained by analyzing the dependence of compound 1 concentration on
the cAMP production elicited by 60 nM of GLP-1. As revealed in FIG.
9C, 60 nM of GLP-1 generated cAMP more than 80% of that of
saturation dose, as the concentration of compound 1 increased, the
cAMP production reduced and reached bottom saturation at 0.003
mg/ml of compound 1. The affinity of compound 1 for GLP-1 in this
assay was measured to be 0.0003 mg/ml. The effect of galanal B,
compound 1 and compound 2 on the affinity of GLP-1 to elicit cAMP
production in RINm5F cells are summarized in Table 3. To examine if
these modulation effects of compound 1 and compound 2 is specific
for GLP-1, 0.03 and 0.01 mg/ml of these compounds were included in
the analysis of dose dependence of GIP and glucagon on cAMP
production in RINm5F cells. As shown in FIG. 11, compound 1 and
compound 2 at a concentration of 0.025 mg/ml do not affect the cAMP
production elicited by GIP or glucagon, indicating their modulation
effects are specific for GLP-1. The above studies revealed that
galanal B, compound 1, and compound 2 are modulator without
intrinsic agonistic or antagonistic activity, their actions are
dependent on both GLP-1 and GLP-1 receptor.
Discussion
TABLE-US-00003 [0217] TABLE 3 Effect of compound 1, 2, and galanal
B on the affinity of GLP-1 dependent cAMP production in RINm5F
cells GLP-1 Compound 1* + Compound 2* + Galanal B* + only GLP-1
GLP-1 GLP-1 EC.sub.50 (nM) 7.86 365 0.0025 4.93 *Titration of GLP-1
on the production of cAMP in RINm5F cells with 0.003 mg/ml of the
indicated compounds.
[0218] Plants remain either the source of or the inspiration for a
significant proportion of the new small-molecule chemical entities.
GLP-1 receptor mediated signaling is a major target for treatment
of type 2 diabetes, further its role in Alzheimer's disease and
psoriasis is under clinical investigation. Currently GLP-1
therapeutics are GLP-1 analogs or compounds with agonistic activity
which may cause serious adverse effect upon chronic use. In an
attempt to find compound that function as a GLP-1 modulator,
instead of agonist, HC plant was identified that displays positive
modulating action on GLP-1 elicited receptor endocytosis which
requires the presence of GLP-1, indicating the identified activity
functions as a GLP-1 potentiator with little intrinsic agonistic
activity. After activity directed fractionation and purification,
the structure of one of the active compounds was found to be
galanal b. The purified galanal b displayed potent activity to
potentiate GLP-1 to elicit recruiting .beta.-arrestin2 to GLP-1
receptor and the following receptor endocytosis. This activity of
galanal b is specific for GLP-1 as it did not show similar effect
on PTH and its cognate receptor. The effect is highly dependent on
the presence of GLP-1 and is abolished by the presence of GLP-1
receptor antagonist exendin 9. To confirm this activity of purified
galanal B, galanal B and its modified analogs were synthesized,
followed by characterizing their ability (galanal b, compound 1,
and compound 2) with respect to potentiating GLP-1 in eliciting
receptor endocytosis and in stimulating cAMP production.
[0219] Though galanal b, compound 1, and compound 2 are all able to
potentiate GLP-1 to elicit receptor endocytosis in a GLP-1 and
GLP-1 receptor dependent manner, their ability to modulate GLP-1 in
stimulating cAMP production is quite distinct, in that compound 1
selectively reduce the affinity of GLP-1 by a factor of 50, galanal
b is neutral, while compound 2 selectively potentiate the affinity
of GLP-1 up to 1000 fold. This finding demonstrates that compound 1
negatively modulate GLP-1 induced cAMP production pathway while
compound 2 positively modulate the same pathway, though they
display comparable activity on GLP-1 elicited receptor endocytosis.
It was demonstrated that these activities and selectivity are
dependent on the GLP-1, GLP-1 receptor and specific for the GLP-1
signaling, as they are GLP-1 concentration dependent, are all
blocked by GLP-1 receptor antagonist exendin 9 and do not affect
cAMP production elicited by the other incretin GIP or by glucagon.
Galanal b, compound 1, and compound 2 share similar scaffolds but
are structurally distinct, the present disclosure identifies a
critical chemical space of galanal b relevant to its selectivity in
modulating GLP-1 receptor coupling efficiency to its intracellular
receptor signaling pathway. The present data show that galanal b,
compound 1, and compound 2 can display quite distinct effect on the
coupling efficiency of GLP-1 receptor to G.alpha.s while display
similar effect on the coupling efficiency to .beta.-arrestin2. The
simplest explanation for this observation is that each compound
will dictate a unique conformation of receptor-GLP-1-compound
complex, thus the conformation of GLP-1 receptor is highly
dependent on whether galanal b, compound 1, or compound 2 is in the
complex. This is consistent with the finding that GPCR can exist in
multiple active conformations, distinct conformation of the
receptor is stabilized by distinct ligand structure and may lead to
distinct signaling selectivity (55). The present findings raise
question as how galanal b and its analogs share similar structure
but stabilize quite distinct conformation of GLP-1 receptor which
coupled comparably to .beta.-arrestin-2 but display opposite
coupling efficiency to the pathway of cAMP production. There are at
least two possible mechanisms to account for these observations.
The first explanation is that the compound may bind to and modify
the structure of GLP-1 receptor such that manifest as a
pathway-dependent change in the signaling capacity upon binding to
its orthosteric ligand GLP-1. By binding to and modifying
structural conformation of the receptor, the allosteric inhibitor
of parturition (PDC 113.824) induces biased signaling when an
orthosteric ligand is co-bound to the prostaglandin F2.alpha.
receptor (56). Alternatively, these compounds bind to GLP-1 to form
a complex with distinct conformation which will stabilize a
distinct set of receptor conformation upon binding and leads to
positively or negatively modulate coupling efficiency to G.alpha.s
pathway while display similarly the coupling efficiency to
.beta.-arrestin2 mediated receptor endocytosis. Subtle conformation
changes in peptide agonists can lead to selective coupling of the
receptor to its downstream signaling pathway. As it has been shown
in the case of biased agonist for the type 1 parathyroid hormone
receptor (PTH1R) (25) and angiotensin II type 1A receptor ligands
(57), subtle structure change of the peptide ligands can profoundly
stabilize a distinct conformation of the receptor protein that
selectively affects the coupling efficiency of a particular
downstream signaling pathway. Two preliminary observations in the
present communication are consistent with the second mechanism that
requires the binding of these compounds to GLP-1 peptide to
stabilize compound-GLP-1 complex in a distinct conformation
different from that of free GLP-1. First, free GLP-1 peptide is
quickly degraded by limited trypsin digestion, while galanal b
protects GLP-1 from trypsin degradation. Furthermore, compound 2
potentiates GLP-1 to stimulate cAMP production in RINm5F cells, but
has little effect on cAMP production by a small molecule agonist
Boc5, though 1000 fold higher of Boc5 is needed to stimulate low
level cAMP production in RINm5F cells.
[0220] The chronic administration of glucagon-like peptide-1
(GLP-1) analogs widely used to treat type-2 diabetes was associated
with a potential risk of pancreatitis (37-41) or pancreatic/thyroid
cancers (42), though with benefits far outweighing the potential
risks (58). Physiologically, plasma level of GLP-1 is stringently
controlled by ingestion of food and by DPP-4, the plasma level of
active GLP-1 will be raised from 5 pM to 20-25 pM 15 min after
glucose challenge and return to basal level 2 hr later. However, a
constant high plasma concentration of GLP-1 analogs in type 2
diabetes receive GLP-1 analogs therapy (59,60) leads to stimulating
target tissues constitutively and may cause the adverse undesired
consequences reported in the literatures. Compound 2 which
potentiate GLP-1 dependent cAMP production by 1000 fold in RINm5F
cells is a "true" positive modulator for GLP-1 because it lacks
intrinsic agonistic activity as it does not elicit cAMP production
in the absence of GLP-1. GLP-1 positive modulator function as a
dimmer switch that amplifies the signaling depends on the plasma
level of endogenous GLP-1, the intensity of activation of GLP-1
receptor is controlled by the physiologic concentration of GLP-1
secreted from intestine, thus will not stimulate target tissues
constitutively. Since GLP-1 secretion in type 2 diabetes is
comparable to or slightly defective as compare to healthy subject
(43,44), GLP-1 positive modulator will be a potential compound to
treat type 2 diabetes in the future. Positive modulator without
intrinsic agonistic activity but only function to potentiate the
activity of endogenous GLP-1 is expected to overcome the undesired
effects by skipping the step of constitutive stimulation of the
target tissues (58) associated with the chronic administration of
GLP-1 analogs.
[0221] GLP-1 receptors are also expressed in extra-pancreatic
tissues, and trial data suggest GLP-1RAs also have effects beyond
their glycaemic actions. GLP-1 signaling has been shown to be
potential target for the treatment of immune dysfunction (Ahern et
al., J Eur Acad Dermatol Venereol. 2013 November; 27(11):1440-3)
(15), neurodegenerative diseases and cardiovascular disorders
(Seufert et al., Diabetes Obes Metab. 2013 Dec. 24. doi:
10.1111/dom.12251; Egefjord et al., Dan Med J. 2012 October;
59(10):A4519). Biological effects triggered by GLP-1 receptor often
result from the activation of multiple intracellular signaling
pathways. Deciphering which signaling pathways are engaged
following GLP-1 receptor activation appears to be primordial to
reveal their contribution in the physiological and pathological
processes. The development of pathway selective GLP-1 modulators to
elucidate the role of the different signaling mechanisms mediated
by GLP-1 receptor activation may allow the generation of new
therapeutic agents with improved efficacy and reduced side effects.
In this regard, the identification of GLP-1 modulator selectively
promoting insulin secretion without inducing pro-inflammatory
effects would offer therapeutic benefit. For many GPCR targets, the
required spectrum of signaling needed to attain optimal therapeutic
benefit is currently unknown, which limits the rational selection
of drug candidates. Therefore, drug discovery at these tractable
targets is considerably challenging. There are evidences showing
that many adverse side effects can be avoided with such pathway
selective compounds and provide improved treatments. The
angiotensin II type 1A receptor, the .beta.-arrestin-biased ligand
Sar1, D-Ala8 angiotensin II (TRV120027) has been shown to increase
cardiac performance in anesthetized rats, whereas unbiased ligands
reduce cardiac performance. There is also potential for improved
PTH receptor agonists used for the treatment of osteoporosis. The
.beta.-arrestin-biased ligand (D-Trp12,Tyr34)-PTH(7-34) stimulates
.beta.-arrestin while blocking G-protein signaling and promotes
anabolic bone formation in the absence of bone resorption.
Screening signaling pathway selective compounds provides
considerable scope for the identification of compounds that
selectively target clinically useful GLP-1 signaling pathways and
are more neutral or even block alternative pathways, which give
rise to undesirable side effects. Given the risk of chronic usage
of GLP-1 agonist in type 2 diabetes and potentially in other
disorders, creating small molecules which modulates GLP-1 signaling
pathway selectivity in a potent unique manner, may dramatically
accelerate the rate at which critical pathway are selected.
Other Embodiments
[0222] All of the features disclosed in this specification may be
combined in any combination. Each feature disclosed in this
specification may be replaced by an alternative feature serving the
same, equivalent, or similar purpose. Thus, unless expressly stated
otherwise, each feature disclosed is only an example of a generic
series of equivalent or similar features.
[0223] From the above description, one skilled in the art can
easily ascertain the essential characteristics of the present
disclosure, and without departing from the spirit and scope
thereof, can make various changes and modifications of the present
disclosure to adapt it to various usages and conditions. For
example, compounds structurally analogous the compounds described
herein of this present disclosure also can be made, screened for
their anti-cancer activities, and used to practice this present
disclosure. Thus, other embodiments are also within the claims.
[0224] All publications and patent applications cited in this
specification are herein incorporated by reference as if each
individual publication or patent application were specifically and
individually indicated to be incorporated by reference for the
purposes or subject matter referenced herein.
REFERENCES
[0225] 1. Bonadonna, R. C., and De Fronzo, R. A. (1991) Glucose
metabolism in obesity and type 2 diabetes. Diabete &
metabolisme 17, 112-135. [0226] 2. Luzi, L., Petrides, A. S., and
De Fronzo, R. A. (1993) Different sensitivity of glucose and amino
acid metabolism to insulin in NIDDM. Diabetes 42, 1868-1877. [0227]
3. Mitri, J., and Hamdy, O. (2009) Diabetes medications and body
weight. Expert opinion on drug safety 8, 573-584. [0228] 4. Roumie,
C. L., Hung, A. M., Greevy, R. A., Grijalva, C. G., Liu, X., Murff,
H. J., Elasy, T. A., and Griffin, M. R. (2012) Comparative
effectiveness of sulfonylurea and metformin monotherapy on
cardiovascular events in type 2 diabetes mellitus: a cohort study.
Annals of internal medicine 157, 601-610. [0229] 5. Drucker, D. J.
(2003) Enhancing incretin action for the treatment of type 2
diabetes. Diabetes care 26, 2929-2940. [0230] 6. Dalle, S.,
Burcelin, R., and Gourdy, P. (2012) Specific actions of GLP-1
receptor agonists and DPP4 inhibitors for the treatment of
pancreatic beta-cell impairments in type 2 diabetes. Cellular
signalling. [0231] 7. Zander, M., Madsbad, S., Madsen, J. L., and
Hoist, J. J. (2002) Effect of 6-week course of glucagon-like
peptide 1 on glycaemic control, insulin sensitivity, and beta-cell
function in type 2 diabetes: a parallel-group study. Lancet 359,
824-830. [0232] 8. Verges, B., Bonnard, C., and Renard, E. (2011)
Beyond glucose lowering: glucagon-like peptide-1 receptor agonists,
body weight and the cardiovascular system. Diabetes &
metabolism 37, 477-488. [0233] 9. Sivertsen, J., Rosenmeier, J.,
Holst, J. J., and Vilsboll, T. (2012) The effect of glucagon-like
peptide 1 on cardiovascular risk. Nature reviews. Cardiology 9,
209-222. [0234] 10. Holst, J. J., Burcelin, R., and Nathanson, E.
(2011) Neuroprotective properties of GLP-1: theoretical and
practical applications. Current medical research and opinion 27,
547-558. [0235] 11. Holscher, C. (2012) Potential role of
glucagon-like peptide-1 (GLP-1) in neuroprotection. CNS drugs 26,
871-882. [0236] 12. Holscher, C. (2013) Central effects of GLP-1:
new opportunities for treatments of neurodegenerative diseases. The
Journal of endocrinology. [0237] 13. Harkavyi, A., and Whitton, P.
S. (2010) Glucagon-like peptide 1 receptor stimulation as a means
of neuroprotection. British journal of pharmacology 159, 495-501.
[0238] 14. Drucker, D. J., and Rosen, C. F. (2011) Glucagon-like
peptide-1 (GLP-1) receptor agonists, obesity and psoriasis:
diabetes meets dermatology. Diabetologia 54, 2741-2744. [0239] 15.
Faurschou, A., Knop, F. K., Thyssen, J. P., Zachariae, C., Skov,
L., and Vilsboll, T. (2011) Improvement in psoriasis after
treatment with the glucagon-like peptide-1 receptor agonist
liraglutide. Acta diabetologica. 2011 Dec. 13. [0240] 16. Hogan, A.
E., Tobin, A. M., Ahem, T., Corrigan, M. A., Gaoatswe, G., Jackson,
R., O'Reilly, V., Lynch, L., Doherty, D. G., Moynagh, P. N., Kirby,
B., O'Connell, J., and O'Shea, D. (2011) Glucagon-like peptide-1
(GLP-1) and the regulation of human invariant natural killer T
cells: lessons from obesity, diabetes and psoriasis. Diabetologia
54, 2745-2754. [0241] 17. Burgmaier, M., Heinrich, C., and Marx, N.
(2013) Cardiovascular effects of GLP-1 and GLP-1-based therapies:
implications for the cardiovascular continuum in diabetes? Diabetic
medicine: a journal of the British Diabetic Association 30,
289-299. [0242] 18. Meloni, A. R., DeYoung, M. B., Lowe, C., and
Parkes, D. G. (2013) GLP-1 receptor activated insulin secretion
from pancreatic beta-cells: mechanism and glucose dependence.
Diabetes, obesity & metabolism 15, 15-27. [0243] 19. Mannucci,
E., and Dicembrini, I. (2012) Incretin-based therapies and
cardiovascular risk. Current medical research and opinion 28,
715-721. [0244] 20. Lefkowitz, R. J., and Shenoy, S. K. (2005)
Transduction of receptor signals by beta-arrestins. Science (New
York, N.Y.) 308, 512-517. [0245] 21. Reiter, E., and Lefkowitz, R.
J. (2006) GRKs and beta-arrestins: roles in receptor silencing,
trafficking and signaling. Trends in endocrinology and metabolism:
TEM 17, 159-165. [0246] 22. Penela, P., Murga, C., Ribas, C.,
Lafarga, V., and Mayor, F., Jr. (2010) The complex G
protein-coupled receptor kinase 2 (GRK2) interactome unveils new
physiopathological targets. British journal of pharmacology 160,
821-832. [0247] 23. Xiao, K., McClatchy, D. B., Shukla, A. K.,
Zhao, Y., Chen, M., Shenoy, S. K., Yates, J. R., 3rd, and
Lefkowitz, R. J. (2007) Functional specialization of beta-arrestin
interactions revealed by proteomic analysis. Proceedings of the
National Academy of Sciences of the United States of America 104,
12011-12016. [0248] 24. Xiao, K., Sun, J., Kim, J., Rajagopal, S.,
Zhai, B., Villen, J., Haas, W., Kovacs, J. J., Shukla, A. K., Hara,
M. R., Hernandez, M., Lachmann, A., Zhao, S., Lin, Y., Cheng, Y.,
Mizuno, K., Ma'ayan, A., Gygi, S. P., and Lefkowitz, R. J. (2010)
Global phosphorylation analysis of beta-arrestin-mediated signaling
downstream of a seven transmembrane receptor (7TMR). Proceedings of
the National Academy of Sciences of the United States of America
107, 15299-15304. [0249] 25. Bohinc, B. N., and Gesty-Palmer, D.
(2012) Biased agonism at the parathyroid hormone receptor: a
demonstration of functional selectivity in bone metabolism. Mini
reviews in medicinal chemistry 12, 856-865. [0250] 26. Kenakin, T.,
and Christopoulos, A. (2013) Signalling bias in new drug discovery:
detection, quantification and therapeutic impact. Nature reviews.
Drug discovery 12, 205-216. [0251] 27. Thomsen, A. R., Smajilovic,
S., and Brauner-Osborne, H. (2012) Novel strategies in drug
discovery of the calcium-sensing receptor based on biased
signaling. Current drug targets 13, 1324-1335. [0252] 28. Garland,
S. L. (2013) Are GPCRs Still a Source of New Targets? Journal of
biomolecular screening 18, 947-966. [0253] 29. DeWire, S. M.,
Yamashita, D. S., Rominger, D. H., Liu, G., Cowan, C. L., Graczyk,
T. M., Chen, X. T., Pitis, P. M., Gotchev, D., Yuan, C., Koblish,
M., Lark, M. W., and Violin, J. D. (2013) A G protein-biased ligand
at the mu-opioid receptor is potently analgesic with reduced
gastrointestinal and respiratory dysfunction compared with
morphine. The Journal of pharmacology and experimental therapeutics
344, 708-717. [0254] 30. Quoyer, J., Longuet, C., Broca, C., Linck,
N., Costes, S., Varin, E., Bockaert, J., Bertrand, G., and Dalle,
S. (2010) GLP-1 mediates antiapoptotic effect by phosphorylating
Bad through a beta-arrestin 1-mediated ERK1/2 activation in
pancreatic beta-cells. The Journal of biological chemistry 285,
1989-2002. [0255] 31. Talbot, J., Joly, E., Prentki, M., and
Buteau, J. (2012) beta-Arrestin1-mediated recruitment of c-Src
underlies the proliferative action of glucagon-like peptide-1 in
pancreatic beta INS832/13 cells. Molecular and cellular
endocrinology 364, 65-70. [0256] 32. Smushkin, G., Sathananthan,
A., Man, C. D., Zinsmeister, A. R., Camilleri, M., Cobelli, C.,
Rizza, R. A., and Vella, A. (2012) Defects in GLP-1 response to an
oral challenge do not play a significant role in the pathogenesis
of prediabetes. The Journal of clinical endocrinology and
metabolism 97, 589-598. [0257] 33. Kreymann, B., Williams, G.,
Ghatei, M. A., and Bloom, S. R. (1987) Glucagon-like peptide-1
7-36: a physiological incretin in man. Lancet 2, 1300-1304. [0258]
34. Orskov, C., Rabenhoj, L., Wettergren, A., Kofod, H., and Holst,
J. J. (1994) Tissue and plasma concentrations of amidated and
glycine-extended glucagon-like peptide I in humans. Diabetes 43,
535-539. [0259] 35. Chiu, W. Y., Shih, S. R., and Tseng, C. H.
(2012) A review on the association between glucagon-like peptide-1
receptor agonists and thyroid cancer. Experimental diabetes
research 2012, 924168. [0260] 36. Butler, P. C., Dry, S., and
Elashoff, R. (2010) GLP-1-based therapy for diabetes: what you do
not know can hurt you. Diabetes care 33, 453-455. [0261] 37. Singh,
S., Chang, H. Y., Richards, T. M., Weiner, J. P., Clark, J. M., and
Segal, J. B. (2013) Glucagonlike peptide 1-based therapies and risk
of hospitalization for acute pancreatitis in type 2 diabetes
mellitus: a population-based matched case-control study. JAMA
internal medicine 173, 534-539. [0262] 38. Gale, E. A. (2013) GLP-1
based agents and acute pancreatitis: drug safety falls victim to
the three monkey paradigm. BMJ (Clinical research ed.) 346, f1263.
[0263] 39. Cohen, D. (2013) Has pancreatic damage from glucagon
suppressing diabetes drugs been underplayed? BMJ (Clinical research
ed.) 346, f3680. [0264] 40. Gale, E. (2013) Incretin therapy:
should adverse consequences have been anticipated? BMJ (Clinical
research ed.) 346, f3617. [0265] 41. Ahmad, S. R., and Swann, J.
(2008) Exenatide and rare adverse events. The New England journal
of medicine 358, 1970-1971; discussion 1971-1972. [0266] 42.
Butler, A. E., Campbell-Thompson, M., Gurlo, T., Dawson, D. W.,
Atkinson, M., and Butler, P. C. (2013) Marked expansion of exocrine
and endocrine pancreas with incretin therapy in humans with
increased exocrine pancreas dysplasia and the potential for
glucagon-producing neuroendocrine tumors. Diabetes 62, 2595-2604.
[0267] 43. Nauck, M. A., Vardarli, I., Deacon, C. F., Holst, J. J.,
and Meier, J. J. (2011) Secretion of glucagon-like peptide-1
(GLP-1) in type 2 diabetes: what is up, what is down? Diabetologia
54, 10-18. [0268] 44. Calanna, S., Christensen, M., Holst, J. J.,
Laferrere, B., Gluud, L. L., Vilsboll, T., and Knop, F. K. (2013)
Secretion of glucagon-like peptide-1 in patients with type 2
diabetes mellitus: systematic review and meta-analyses of clinical
studies. Diabetologia 56, 965-972. [0269] 45. Barak, L. S.,
Ferguson, S. S., Zhang, J., and Caron, M. G. (1997) A
beta-arrestin/green fluorescent protein biosensor for detecting G
protein-coupled receptor activation. The Journal of biological
chemistry 272, 27497-27500. [0270] 46. Pierce, K. L., and
Lefkowitz, R. J. (2001) Classical and new roles of beta-arrestins
in the regulation of G-protein-coupled receptors. Nature reviews.
Neuroscience 2, 727-733. [0271] 47. Ponsioen, B., Zhao, J., Riedl,
J., Zwartkruis, F., van der Krogt, G., Zaccolo, M., Moolenaar, W.
H., Bos, J. L., and Jalink, K. (2004) Detecting cAMP-induced Epac
activation by fluorescence resonance energy transfer: Epac as a
novel cAMP indicator. EMBO reports 5, 1176-1180. [0272] 48. Cheng,
H. F., Jiang, M. J., Chen, C. L., Liu, S. M., Wong, L. P.,
Lomasney, J. W., and King, K. (1995) Cloning and identification of
amino acid residues of human phospholipase C delta 1 essential for
catalysis. The Journal of biological chemistry 270, 5495-5505.
[0273] 49. Hsu, Y. Y., Liu, Y. N., Lu, W. W., and Kung, S. H.
(2009) Visualizing and quantifying the differential cleavages of
the eukaryotic translation initiation factors eIF4GI and eIF4GII in
the enterovirus-infected cell. Biotechnology and bioengineering
104, 1142-1152. [0274] 50. Chick, W. L., Warren, S., Chute, R. N.,
Like, A. A., Lauris, V., and Kitchen, K. C. (1977) A transplantable
insulinoma in the rat. Proceedings of the National Academy of
Sciences of the United States of America 74, 628-632. [0275] 51.
Ghosh, R. N., DeBiasio, R., Hudson, C. C., Ramer, E. R., Cowan, C.
L., and Oakley, R. H. (2005) Quantitative cell-based high-content
screening for vasopressin receptor agonists using transfluor
technology. Journal of biomolecular screening 10, 476-484. [0276]
52. Hudson, C. C., Oakley, R. H., Sjaastad, M. D., and Loomis, C.
R. (2006) High-content screening of known G protein-coupled
receptors by arrestin translocation. Methods in enzymology 414,
63-78. [0277] 53. Jiang, L. I., Collins, J., Davis, R., Lin, K. M.,
DeCamp, D., Roach, T., Hsuch, R., Rebres, R. A., Ross, E. M.,
Taussig, R., Fraser, I., and Sternweis, P. C. (2007) Use of a cAMP
BRET sensor to characterize a novel regulation of cAMP by the
sphingosine 1-phosphate/G 13 pathway. The Journal of biological
chemistry 282, 10576-10584. [0278] 54. Nikolaev, V. O., Bunemann,
M., Hein, L., Hannawacker, A., and Lohse, M. J. (2004) Novel single
chain cAMP sensors for receptor-induced signal propagation. The
Journal of biological chemistry 279, 37215-37218. [0279] 55.
Reiter, E., Ahn, S., Shukla, A. K., and Lefkowitz, R. J. (2012)
Molecular mechanism of beta-arrestin-biased agonism at
seven-transmembrane receptors. Annual review of pharmacology and
toxicology 52, 179-197. [0280] 56. Goupil, E., Tassy, D., Bourguet,
C., Quiniou, C., Wisehart, V., Petrin, D., Le Gouill, C., Devost,
D., Zingg, H. H., Bouvier, M., Saragovi, H. U., Chemtob, S.,
Lubell, W. D., Claing, A., Hebert, T. E., and Laporte, S. A. (2010)
A novel biased allosteric compound inhibitor of parturition
selectively impedes the prostaglandin F2alpha-mediated Rho/ROCK
signaling pathway. The Journal of biological chemistry 285,
25624-25636. [0281] 57. Violin, J. D., DeWire, S. M., Yamashita,
D., Rominger, D. H., Nguyen, L., Schiller, K., Whalen, E. J.,
Gowen, M., and Lark, M. W. (2010) Selectively engaging
beta-arrestins at the angiotensin II type 1 receptor reduces blood
pressure and increases cardiac performance. The Journal of
pharmacology and experimental therapeutics 335, 572-579. [0282] 58.
Butler, P. C., Elashoff, M., Elashoff, R., and Gale, E. A. (2013) A
critical analysis of the clinical use of incretin-based therapies:
Are the GLP-1 therapies safe? Diabetes care 36, 2118-2125. [0283]
59. Kim, D., MacConell, L., Zhuang, D., Kothare, P. A., Trautmann,
M., Fineman, M., and Taylor, K. (2007) Effects of once-weekly
dosing of a long-acting release formulation of exenatide on glucose
control and body weight in subjects with type 2 diabetes. Diabetes
care 30, 1487-1493. [0284] 60. Calara, F., Taylor, K., Han, J.,
Zabala, E., Carr, E. M., Wintle, M., and Fineman, M. (2005) A
randomized, open-label, crossover study examining the effect of
injection site on bioavailability of exenatide (synthetic
exendin-4). Clinical therapeutics 27, 210-215.
* * * * *