U.S. patent application number 17/129678 was filed with the patent office on 2021-06-17 for utility of (+) epicatechin and their analogs.
The applicant listed for this patent is Epirium Bio Inc.. Invention is credited to Sundeep DUGAR, George SCHREINER, Somdutta SEN.
Application Number | 20210177801 17/129678 |
Document ID | / |
Family ID | 1000005421519 |
Filed Date | 2021-06-17 |
United States Patent
Application |
20210177801 |
Kind Code |
A1 |
DUGAR; Sundeep ; et
al. |
June 17, 2021 |
UTILITY OF (+) EPICATECHIN AND THEIR ANALOGS
Abstract
The present invention pertains to the enhanced activity of (+)
epicatechin over (-) epicatechin. The present invention is related
to novel analogs of (+) epicatechin of the formula (I), which
enhances the pharmacokinetics and therefore the pharmacodynamics of
(+) epicatechin. The present invention is related to analogs of (+)
epicatechin of the formula (I). The general structure of the
analogs of the present invention may be represented by Formula (I):
Formula (I) wherein A and B are independently OR1 and C and D are
independently OH; wherein R1 is independently C1 to C10 lower
straight or branched chain acyclic or cyclic alkyl, or is selected
from the group comprising, hydroxy butyric acid, dichloroacetic
acid; phenyl butyric acid; valproic acid.
Inventors: |
DUGAR; Sundeep; (San Jose,
CA) ; SCHREINER; George; (Los Altos Hills, CA)
; SEN; Somdutta; (Manesar, IN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Epirium Bio Inc. |
San Diego |
CA |
US |
|
|
Family ID: |
1000005421519 |
Appl. No.: |
17/129678 |
Filed: |
December 21, 2020 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
16311422 |
Dec 19, 2018 |
10898465 |
|
|
PCT/IN2017/050252 |
Jun 21, 2017 |
|
|
|
17129678 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 1/16 20180101; A61P
25/28 20180101; A61K 31/353 20130101; C07D 311/62 20130101; A61P
9/10 20180101; A61P 21/00 20180101; A61P 25/08 20180101 |
International
Class: |
A61K 31/353 20060101
A61K031/353; C07D 311/62 20060101 C07D311/62; A61P 21/00 20060101
A61P021/00; A61P 9/10 20060101 A61P009/10; A61P 25/28 20060101
A61P025/28; A61P 1/16 20060101 A61P001/16; A61P 25/08 20060101
A61P025/08 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 21, 2016 |
NL |
4200/DEL/2015 |
Jun 22, 2016 |
NL |
201611021674 |
Claims
1.-7. (canceled)
8. A method for treating a disease or a disorder associated with
Electron Transport Chain, comprising administering to a subject in
need thereof a therapeutically effective amount of (+)-epicatechin
or an analog of (+) epicatechin of Formula (I), ##STR00025##
wherein A and B are independently OR.sup.1 and C and D are
independently OH, or B is OR.sup.1 and A, C and D are independently
OH; wherein R.sup.1 is independently C.sub.2 to C.sub.10 lower
straight or branched chain acyclic or cyclic alkyl, or
--C(.dbd.O)--(C.sub.6-C.sub.9 alkyl), or, taken together with the
oxygen to which it is attached, is selected from the group
consisting of hydroxybutanoate, dichloroacetate, phenyl butanoate,
phenyl propionate, and 2-propylpentanoate.
9. The method of claim 8, wherein B is OR.sup.1 and A, C and D are
independently OH.
10. The method of claim 8, wherein A and B are independently
OR.sup.1 and C and D are independently OH.
11. The method of claim 8, wherein R.sup.1, taken together with the
oxygen to which it is attached, is selected from the group
consisting of hydroxybutanoate, dichloroacetate, phenyl butanoate,
phenyl propionate, and 2-propylpentanoate.
12. The method of claim 8, wherein R.sup.1 is
--C(.dbd.O)--(C.sub.6-C.sub.9 alkyl).
13. The method of claim 8, wherein the analog of (+) epicatechin is
selected from the group comprising: i.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,7-dihydroxychroman-5-yl
octanoate; ii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,5-dihydroxychroman-7-yl
octanoate; iii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3-hydroxychroman-5,7-diyl
dioctanoate; iv.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,7-dihydroxychroman-5-yl
heptanoate; v.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,5-dihydroxychroman-7-yl
heptanoate; vi.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3-hydroxychroman-5,7-diyl
diheptanoate; vii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,7-dihydroxychroman-5-yl
decanoate; viii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,5-dihydroxychroman-7-yl
decanoate; ix.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3-hydroxychroman-5,7-diyl
bis(decanoate); x.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3-hydroxychroman-5,7-diylbis(2-propylpent-
anoate); xi.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,7-dihydroxychroman-5-yl
2-propylpentanoate; xii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,5-dihydroxychroman-7-yl
2-propylpentanoate; xiii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3-hydroxy-5-((3-phenylpropanoyl)oxy)chrom-
an-7-yl 4-phenylbutanoate; xiv.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,7-dihydroxychroman-5-yl
3-phenylpropanoate; xv.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,5-dihydroxychroman-7-yl
4-phenylbutanoate; xvi.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3-hydroxychroman-5,7-diyl
bis(2,2-dichloroacetate); xvii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,7-dihydroxychroman-5-yl
2,2-dichloroacetate; and xviii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,5-dihydroxychroman-7-yl
2,2-dichloroacetate.
14. The method of claim 8, wherein the disease or the disorder is
associated with Electron Transport Chain, complex IV.
15. The method of claim 8, wherein the disease or the disorder is
selected from the group consisting of impaired cognition, a
neurodegenerative disease, dystonia, sarcopenia, cardiomyopathy of
aging or other diseases associated with mitochondrial dysfunction,
ischemic vascular disease, immunodeficiency states, ataxia,
pulmonary inflammation and fibrosis, infantile encephalomyopathy,
epilepsy, Charcot-Marie-Tooth disease, exocrine pancreatic
insufficiency, impaired wound healing, and growth of cancer
cells.
16. The method of claim 15, wherein the neurodegenerative disease
is Alzheimer's or Leigh syndrome.
17. The method of claim 8, wherein the (+)-epicatechin is free of
catechin and other isomers of epicatechin.
Description
FIELD OF THE INVENTION
[0001] The present invention discloses the utility of (+)
epicatechin and analogs of (+) isoform of epicatechin.
BACKGROUND OF INVENTION
[0002] Polyphenolic natural products are important because of their
utility in various biological pathways, their occurrence in
foodstuffs, and hence their relevance for human health. The
stereochemistry of the substituents on a polyphenol monomeric unit
of a polyphenol may be described in terms of their relative
stereochemistry, "alpha/beta" or "cis/trans". The term "alpha" ( )
indicates that the substituent is oriented above the plane of the
flavanol ring, whereas, "beta" ( ) indicates that the substituent
is oriented above the plane of the ring. The term "cis" indicates
that two substituents are oriented on the same face of the ring,
whereas "trans" indicates that two substituents are oriented on
opposite faces of the ring.
[0003] Catechins possess two benzene rings and a dihydropyran
heterocycle (the C-ring) with a hydroxyl group on carbon 3. A ring
is similar to a resorcinol moiety while the B ring is similar to a
catechol moiety. There are two chiral centers on the molecule, on
carbons 2 and 3. Therefore, it has four diastereoisomers. Two of
the isomers are in a trans configuration and are called catechin
and the other two are in a cis configuration and are called
epicatechin.
[0004] (+)-Catechin and (-)-epicatechin are the most abundant
naturally occurring epimers in cacao. During biosynthesis catechin
and epicatechin are predominantly synthesized as (+)-catechin and
(-)-epicatechin. However, certain plants such as spotted knapweed
(Centaurea maculosa, Lam.) demonstrate the presence of racemic
catechin and both (+/-)-catechin and (+/-)-epicatechin was
described in guarana seeds (Paullinia cupana var. sorbilis).
[0005] Since, catechin and epicatechin possess two chiral centers;
their properties depend on the conformation of the molecules. Since
(-)-epicatechin, is the predominantly synthesized or available
epimer of epicatechin in cacao or tea, most of the reports of the
biological activity tested are for this isomer. The activity of
epicatechins as their individual epimers and/or racemic mixture is
not well documented in prior art. The prior art discloses that
naturally occurring member of the flavonoid family, (-)-epicatechin
as inducing mitochondrial biogenesis in vitro and in vivo,
resulting in the successful treatment of diseases associated with
mitochondrial depletion, such as muscular dystrophy. Numerous
papers and patents discuss the broad use of flavonoids as
anti-oxidants or anti-cancer agents. Those teaching this art do not
distinguish chiral flavonoids as being uniquely active. Nor has
there been any report of stereo selective properties of a unique
flavonoid. Rather their effects have typically been described as
attributable to all members of the flavonoid class. There are
reports in prior art that state that (-) isoform of epicatechin,
found naturally occurring in cocoa, green tea, and other plant
sources of polyphenols, can prevent acute mitochondrial injury
involving the formation of mitochondrial permeability transition
pores that damage mitochondrial function by allowing the
non-specific diffusion of electrolytes into the mitochondria and
that (-)-isoform of epicatechin is capable of inducing
mitochondrial biogenesis in in vivo models (see WO 2012/170430 and
WO 2013/142816). The differences in the activities between the
isomers are also not known and documented in the prior art.
[0006] Isolation and availability of pure polyphenols from natural
sources is difficult with increasing degree of oligomerization and
has been one of the reasons for the lack of information about the
stereochemical difference of activity of the enantiomers of
epicatechin and therefore synthesis of polyphenols is preferred. In
addition, such polyphenols have certain drawbacks, when used
clinically, such as, poor pharmacokinetic profile. Hence there is a
need to improve the pharmacokinetic profiles of the
polyphenols.
[0007] One of the consequences of a means of production of a
synthetic epimer is the ability to construct new chemical analogues
of a stereochemically defined phenol. The analogues of polyphenols
may be used, to improve the pharmacokinetic profile of the
polyphenol by, increasing the half-life of the parent drug, which
would help decrease the number of doses needed to achieve a desired
effect, and/or create a more effective and/or a safer drug.
[0008] There are certain prior art drawn to the analogs of
epicatchin, WO 2014/162320, the Applicant disclosed certain novel
analogs of natural flavonoid phenols, that were biologically
active, but the Application, neither discloses the importance of
stereoisoforms in activity, nor does the application disclose the
mode of activity of these analogs.
[0009] Hence, there is a need to examine the utility of the isomers
of catechin/epicatechin and also for novel analogs of epicatechin
that effectively delivers the preferred isomer.
OBJECT OF THE INVENTION
[0010] An object of the invention is to examine the utility of the
isomers of epicatechin and also to provide novel analogs of
epicatechin that effectively delivers the preferred isomer
BRIEF DESCRIPTION OF THE DRAWINGS
[0011] FIG. 1: depicts the activity of (+)-epicatechin in
inhibition complex IV on the increase of the expression of Electron
Transport Chain TV (ETC TV) in comparison to (-) epicatechin,
(+)-epicatechin is approximately 400 fold more potent than
(-)-epicatechin--an unprecedented gain of biological potency.
[0012] FIG. 2: depicts the greater homology to
11-beta-hydroxypregnenolone of (+)-epicatechin compared to that of
(-)-epicatechin.
[0013] FIG. 3: depicts the activity of the compounds on
triglycerides content of livers.
BRIEF DESCRIPTION
[0014] The present invention pertains to the enhanced activity of
(+) epicatechin over (-) epicatechin.
[0015] The present invention is related to novel analogs of (+)
epicatechin of the formula (I), which enhances the pharmacokinetics
and therefore the pharmacodynamics of (+) epicatechin.
[0016] The present invention is related to analogs of (+)
epicatechin of the formula (I). The general structure of the
analogs of the present invention may be represented by Formula
(I):
##STR00001##
wherein A and B are independently OR.sub.1 and C and D are
independently OH; wherein R.sup.1 is independently C.sub.1 to
C.sub.10 lower straight or branched chain acyclic or cyclic alkyl,
or is selected from the group comprising, hydroxy butyric acid.
dichloroacetic acid; phenyl butyric acid; valproic acid.
[0017] The present invention discloses analogs of (+) epicatechin
of the formula (I), wherein B is OR.sub.1 and A, C and D me
independently OH: wherein R.sup.1 is independently C.sub.1 to
C.sub.10 lower straight or branched chain acyclic or cyclic alkyl,
or is selected from the group comprising, L-Glutamic acid, hydroxy
butyric acid, dichloroacetic acid; phony butyric acid; valproic
acid.
[0018] The present invention includes a process for preparation of
compounds of the present invention and methods of use comprising
the compounds of the present invention.
DETAILED DESCRIPTION
[0019] The present invention is based on the unexpected stereo
selectivity with respect to the isomers of epicatechin, which has
two enantiomers. (-)-epicatechin, and (+)-epicatechin.
A. Activity of (+) Epicatechin Compared to (-)Epicatechin
[0020] The physical and biochemical properties of stereo isomers
can differ significantly and unexpectedly. Enantiomers can differ
with respect to activity and physicochemical properties. Stereo
selective metabolism of chiral compounds can influence
pharmacokinetics, pharmacodynamics, and toxicity. There is no
predictability with respect to differential expression of
therapeutic or adverse effects among enantiomers (Agranat I et al
2002 Putting chirality to work: the strategy of chiral switches.
Nature Reviews Drug Discovery 1:753-768; When one enantiomer has
activity of interest, its paired enantiomer typically is either
inactive, or an antagonist of the active enantiomer, or has a
separate activity that could be undesirable. There is no way to
predict or anticipate such outcomes for any given enantiomer
(Caldwell, J, 1999, Through the looking glass in chiral
development. Modern Drug Discov 2:51-60). Occasionally both
enantiomers may show similar activities to varying degrees. It is
more usual to see the greatest degree of variability among the
enantiomers of receptor antagonists, as there are many potential
ways to sterically obstruct the active site of a receptor. The
largest therapeutic variation in potency that we have been able to
determine among enantiomers, therefore, are receptor antagonists
For example, S (-)-propranol exhibits 100-fold greater receptor
antagonism than the R-(+)-propranolol with respect to blocking the
1, 2, and 3 adrenergic receptors. (Smith, S, 2009, Chiral
toxicology; it's the same only different ToxicolSci 110:4-30). The
more restricted requirement of optimal ligand fit to a receptor to
activate the receptor normally results in much smaller variation
with respect to potency of receptor activation. When paired
enantiomers exhibit similar agonist activity, the differences in
potency are typical those of a fractional ratio. The prior art does
not disclose any examples of differential agonist activity of
enantiomers of more than a few fold.
[0021] The present invention discloses a remarkable range of
biological activity across the two enantiomers of epicatechin,
something heretofore not described for flavonoids as a class. The
enantiomer of (-)-epicatechin is (+)-epicatechin. When compared in
an assay on the increase of the expression of Electron Transport
Chain IV (ETC IV), (+)-epicatechin is approximately 400 fold more
potent than (-)-epicatechin--an unprecedented gain of biological
potency (FIG. 1). The data is represented at Table 1:
TABLE-US-00001 TABLE 1 EC.sub.50 (mM) OF COMPOUNDS ON MITOCHONDRIAL
ETC COMPLEXES Compound ELCTRON TRANSPORT CHAIN COMPLEX IV
(-)-Epicatechin 0.04 (+)-Epicatechin 0.0001
[0022] The basis for the advantageous properties of the (-)- and
(+)-isoforms of epicatechin consists of their structural homology
to recently discovered hormone that mediates which is set out in
the patent application PCT/IN2015/050072:
##STR00002##
[0023] As shown in FIG. 2, (-)-epicatechin possesses two hydroxyl
groups that are a steric match for two of the three hydroxyl groups
of OHP. However, when (+)-epicatechin is inverted in relationship
to (-)-epicatechin and matched against OHP, now all 3 of the
hydroxyl groups of OHP display remarkable homology to 3 of the
hydroxyl groups of (+)-epicatechin. There is no precedent for the
discovery that the inverted 3-dimensional structure of one
enantiomer of a compound possesses closer structural and functional
homology to a natural ligand when compared to its paired
enantiomer.
[0024] Therefore, the preferred enantiomer of` epicatechin for use
is the (+) isoform or the (2S,3S) enantiomer of epicatechin and its
analogs, preferably free of contamination with catechin.
(+)-Epicatechin results in a superior pharmacological effect when
free from other flavonoids, particularly from known isomers of
epicatechin.
[0025] Without being limited by theory, it is submitted that the
compounds of the present invention are active due to their unique
configuration and stereochemistry. The compounds of the present
invention are useful in treating diseases or disorders that would
benefit from modification of Electron transfer Chain (ETC) and
particularly electron transfer chain IV.
[0026] The present invention provides methods for treating diseases
or disorders that would benefit from increased expression of
Electron transfer Chain, particularly ETC IV. The methods involve
administering to a subject in need thereof a therapeutically
effective amount of a (+)-epicatechin.
[0027] The vast majority of the body's need for ATP is supplied
through the process of oxidative phosphorylation, carried out in
the mitochondria in all tissues. There are 5 protein complexes,
known as the Electron Transport Complexes that effect ATP
synthesis. ETC I, II, III and IV mediate electron transport. ETC I,
III and IV also function as proton pumps that maintain an
electrochemical gradient necessary for activity of ETC V, the ATP
synthase enzyme that makes ATP from ADP. Complex IV, also known as
cytochrome c oxidase, (COX), consists of 14 subunits whose assembly
into a functional complex requires an additional 30 protein
factors. ETC IV is particularly important to oxidative
phosphorylation. It is the only one of the ETC complexes to
manifest tissue-specific and developmentally regulated isoforms,
allowing precise regulation of oxidative phosphorylation under a
variety of metabolic demands. Thus the ETC IV (COX) protein complex
is considered to be the rate-limiting step in oxidative
phosphorylation. Small positive or negative changes in ETC IV can
exert a significant impact on health, Selective activation of COX
activity has been associated with improved cognition, improved
neuronal cell survival under stress, and improved wound healing.
Mutations in the numerous proteins that comprise or regulate the
activity of ETC IV reveal the pathological consequences of even
modest decreases in ETC IV activity. As little as a 30% reduction
in COX activity has been shown to induce cardiomyopathy or be
associated with the development of neurodegenerative diseases such
as Alzheimer's. Decreases in COX (ETC IV) expression due to
mutations or molecular manipulation have been associated with loss
of muscle endurance and speed, muscle dystonia, immunodeficiency
states due to impaired T cell maturation, cardiomyopathy,
particularly of the aging phenotype, ataxia, neurodegeneration,
increased toxicity in the setting of ischemia, pulmonary
inflammation and fibrosis, encephalopathy, vascular insufficiency,
and stimulation of cancer cell proliferation. Additional specific
diseases associated with COX subunit isoform mutations causing loss
of function include exocrine pancreatic insufficiency, inflammatory
lung disease, Charcot-Marie-Tooth disease, infantile
encephalomyopathy, and Leigh syndrome neurodegeneration with
epilepsy.
[0028] In summary, the following conditions associated with loss of
COX expression or function would be expected to be therapeutically
responsive to a potent, preferential inducer of COX (ETC IV)
expression: impaired cognition, neurodegenerative diseases such as
Alzheimer's or Leigh syndrome, dystonia, sarcopenia, cardiomyopathy
of aging or other diseases associated with mitochondrial
dysfunction, ischemic vascular disease, immunodeficiency states,
ataxia, pulmonary inflammation and fibrosis, infantile
encephalomyopathy, epilepsy. Charcot-Marie-Tooth disease, exocrine
pancreatic insufficiency, impaired wound healing, growth of cancer
cells.
[0029] In addition, given the relative effect of (+)-epicatechin
compared to (-)-epicatechin in lowering the elevated triglycerides
of mice on a high fat diet, (+)-epicatechin and its analogs would
be the preferred medicament for conditions associated with elevated
triglycerides, such as metabolic syndrome, Type II diabetes,
congenital hyperlipidemias, and drug-induced hyperlipidemia, as is
observed with corticosteroid treatments.
B. Analogs of (+) Epicatechin with Increased Pharmacokinetic
Property and Enhanced Utility.
[0030] In another aspect, the present application also discloses
compounds of formula (I) that are analogs of (+)-epicatechin that
possess improved pharmacokinetic properties and enhanced
utility.
[0031] The general structure of the analogs of the present
invention may be represented by Formula (I):
##STR00003##
[0032] Formula (I) wherein A and B are independently OR1 and C and
D are independently OH; wherein R.sup.1 is independently C.sub.1 to
C.sub.10 lower straight or branched chain acyclic or cyclic alkyl,
or is selected from the group comprising, hydroxy butyric acid,
dichloroacetic acid; phenyl butylic add; valproic acid.
[0033] The present invention discloses analogs of (+) epicatechin
of the formula (I), wherein B is OR.sub.1 and A, C and D are
independently OH; wherein R.sup.1 is independently C.sub.1 to
C.sub.10 lower straight or branched chain acyclic or cyclic alkyl,
or is selected from the group comprising, L-Glutamic acid, hydroxy
butyric acid, dichloroacetic acid; phenyl butyric acid; valproic
acid.
[0034] A few illustrative compounds of the present invention are
listed at Table 2.
TABLE-US-00002 TABLE 2 Illustrative Compounds of the Present
Invention. S. No. Structure IUPAC Name 1001 ##STR00004##
(2S,3S)-2-(3,4-dihydroxypheny1)-3,7- dihydroxychroman-5y1 octanoate
1002 ##STR00005## (2S,3S)-2-(3,4-dihydroxypheny1)-3,5-
dihydroxychroman-7-y1 octanoate 1003 ##STR00006##
(2S,3S)-2-(3,4-dihydroxypheny1)- 3-hydroxychroman-5,7-diy1
dioctanoate 1004 ##STR00007## (2S,3S)-2-(3,4-dihydroxyphenyl
3,7-dihydroxychroman-5-y1 heptanoate 1005 ##STR00008##
(2S,3S)-2-(3,4-dihydroxypheny1)- 3,5-dihydroxychroman-7-y1
heptanoate 1006 ##STR00009## (2S,3S)-2-(3,4-dihydroxyphenyl)-
3-hydroxychroman-5,7-diy1 diheptanoate 1007 ##STR00010##
(2S,3S)-2-(3,4-dihydroxyphenyl)- 3,7-dihydroxychroman-5-y1
decanoate 1008 ##STR00011## (2S,3S)-2-(3,4-dihydroxypheny1)-
3,5-dihydroxychroman-7-y1 decanoate 1009 ##STR00012##
(2S,3S)-2-(3,4-dihydroxypheny1)- 3-hydroxychromane-5,7-diy1
bis(decanoate) 1010 ##STR00013## (2S,3S)-2-(3,4-dihydroxypheny1)-
3-hydroxychroman-5,7-diyl bis(2- propyldecanoate) 1011 ##STR00014##
(2S,3S)-2-(3,4-dihydroxypheny1)- 3,7-dihydroxychroman-5-y1 2-
propylpentanoate 1012 ##STR00015## (2S,3S),2 3,4-dihydroxypheny1)-
3,5-dihydroxychroman-7-y1 2- propylpentanoate 1013 ##STR00016##
(2S,3S)-2-(3,4-dihydroxyphenyl)- 3-hydroxy-5-((3-
phenylpropanoyl)oxy)chroman-7- yl 4-phenylbutanoate 1014
##STR00017## (2S,3S)-2-(3,4-dihydroxypheny1)-
3,7-dihydroxychrotnan-5-yl 3- phenylpropanoate 1015 ##STR00018##
(2S,3S)-2-(3,4-dihydroxypheny1)- 3,5-dihydroxychroman-7-y1 4-
phenylbutanoate 1016 ##STR00019## (2S,3S)-2-(3,4-dihydroxypheny1)-
3-hydroxychroman-5,7-diyl bis(2,2-dichloroaceate) 1017 ##STR00020##
(2S,3S)-2-(3,4-dihydroxyphenyl)- 3,7-dihydroxychroman-5-y1
2,2-dichloroacetate 1018 ##STR00021##
(2S,3S)-2-(3,4-dihydroxypheny1)- 3,5-dihydroxychroman-7-y1 2,2-
dichloroacetate
[0035] The compounds of the present invention include: [0036] i.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,7-dihydroxychroman-5-yl
octanoate; [0037] ii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,5-dihydroxychroman-7-yl
octanoate; [0038] iii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3-hydroxychroman-5,7-diyl
dioctanoate; [0039] iv.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,7-dihydroxychroman-5-yl
heptanoate; [0040] v.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,5-dihydroxychroman-7-yl
heptanoate; [0041] vi.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3-hydroxychroman-5,7-diyl
diheptanoate; [0042] vii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,7-dihydroxychroman-5-yl
decanoate; [0043] viii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,5-dihydroxychroman-7-yl
decanoate; [0044] ix.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3-hydroxychroman-5,7-diyl
bis(decanoate); [0045] x.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3-hydroxychroman-5,7-diyl
bis(2-propylpentanoate); [0046] xi.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,7-dihydroxychroman-5-yl
2-propylpentanoate; [0047] xii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,5-dihydroxychroman-7-yl
2-propylpentanoate; [0048] xiii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3-hydroxy-5-((3-phenylpropanoyl)oxy)chrom-
an-7-yl 4-phenylbutanoate; [0049] xiv.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,7-dihydroxychroman-5-yl
3-phenylpropanoate; [0050] xv.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,5-dihydroxychroman-7-yl
4-phenylbutanoate; [0051] xvi.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3-hydroxychroman-5,7-diyl
bis(2,2-dichloroacetate) [0052] xvii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,7-dihydroxychroman-5-yl
2,2-dichloroacetate; [0053] xviii.
(2S,3S)-2-(3,4-dihydroxyphenyl)-3,5-dihydroxychroman-7-yl
2,2-dichloroacetate.
C. Synthesis of the Compounds of the Present Invention.
[0054] The present invention also relates to a process of preparing
the compounds of formula (I). The compounds of present invention
may be prepared by the synthetic scheme 1 as here below:
##STR00022##
[0055] Some of the compound of present interest can be synthesized
from (+)-epicatechin (1) by the scheme outline as above. The (+)
isomer of epicatechin can be synthesized as mentioned in
PCT/IN2012/000052, PCT/IN2014/000061, which are incorporated herein
in its entirety. The (+) isomer of a polyphenol e.g. epicatechin
when treated with a defined quantity of corresponding acylchloride
or carbonyl chloride or carbamoylchlorode in presence of base such
as DIPEA or TEA or potassium carbonate in a suitable solvent such
as acetonitrile or dichloromethane at a temperature range from
0.degree. C. to refluxing can provide substituted derivatives of
interests represented by compound 2.
[0056] In other case, a (+) polyphenol such as (+)-epicatechin can
be protected using a protecting group known in literature such as
CBZ--Cl in presence of a base such as TEA in a solvent such as
acetonitrile at temperature ranging from 0.degree. C. to refluxing
to give the compound represented by 3. Compound 3 can be
derivatized, using different ratios of derivatizing agents to
generate analogs with variable R.sub.1 as defined above using a
base like TEA or DIPEA in a solvent such as acetonitrile at
temperature ranging from 0.degree. C. to refluxing to give analogs
represented by 4, 6 and 8. Subsequent removal of the CBZ groups of
compounds 4, 6 and 8 can give the compounds represented by
structures 5, 7 and 9.
[0057] The present invention discloses methods involve
administering (+)-epicatechin, analogs of (+) epicatechin as set
out herein, and chemical derivatives thereof. The present invention
discloses diseases and disorders that would benefit from increased
mitochondrial activity include diseases or disorders associated
with mitochondrial dysfunction.
[0058] Without being limited by theory, the compounds of the
present invention exhibit superior pharmacokinetic and
pharmacodynamic properties in comparison to (+) epicatechin.
[0059] The present specification is described by way of certain
examples mean for illustration. The examples may not be construed
to limit the scope of the invention in any manner.
Example 1: Synthesis of Compounds of the Present Invention
##STR00023##
[0061] Step-1:
[0062] To a stirred solution of [1] (0.4 gm, 1.379 mmol) in
acetonitrile (40 ml) was added triethylamine (0.38 ml, 1.75 mmol)
followed by benzyl chloroformate (0.39 ml, 2.75 mmol) at 0.degree.
C. under nitrogen atmosphere and stirring at this temperature for
90 mins. Reaction was monitored by TLC, three new spots were
observed along with the starting compound [1]. Reaction mixture was
quenched with NH.sub.4Cl solution (5 ml) and extracted with ethyl
acetate (2.times.50 ml), The combined organic layer was washed with
water, brine and dried over sodium sulphate. The organic layer was
evaporated to afford a light brown solid. This crude product was
loaded on to silica gel column and eluted with 10% ethyl
acetate/hexane to obtain[3] (0.43 gm, 59%) and [10].
[0063] Step-2:
[0064] To a stirred solution of [3] (0.4 gm, 0.766 mmol) in
acetonitrile (40 ml) was added triethylamine (0.105 m, 0.766 mmol)
followed by octanoykhloride (0.124 ml, 0.727 mmol) at 0.degree. C.
under nitrogen atmosphere and stirring at this temperature for 45
mins. Reaction was monitored by TLC. Reaction mixture was quenched
with water (5 ml) and extracted with ethyl acetate (2.times.50 ml).
The combined organic layer was washed with water, brine and dried
over sodium sulphate. The organic layer was evaporated to afford a
light brown solid. This crude product was loaded on to silica gel
column and eluted with 10% ethyl acetate/hexane to off-white powder
[11] (0.110 gm, 22%), [12] and [13].
##STR00024##
[0065] Step-3:
[0066] To a stirred solution of [11] (0.050 g. 0.23 mmol) in ethyl
acetate (10 ml). was added 10% Pd(OH).sub.2 (0.015 g) and stirred
under hydrogen atmosphere at room temperature. The reaction mass
was filtered over celite and the solvent was evaporated out to
afford light yellow sticky material. This crude product was
triturated with ethyl acetate in-pentane to afford yellow sticky
material as [14] (0.025 gm, 80%). Compounds 12 and 13 were
converted to compounds 15 and 16.
Example 2: Effect of (+) Epicatechin on Triglyceride Level
[0067] Animals were placed on High Fat Diet (HFD) until they gain
more than 20% of Body weight compared with animals on standard chow
and reached glycemia levels .gtoreq.200 mg/dL (usually 4-6 weeks).
Animals were randomly assigned to Control (obese group) receiving
vehicle only (by gavage): n=12; (+)-Epicatechin--orally by by
gavage: n=10: (-)-Epicatechin--orally by gavage: n=10.
[0068] All animals were treated for 15 days and continued under
HFD. The results are presented at FIG. 3. Effect on Triglycerides:
(+)-Epicatechin (Dose: 0.003 mg/Kg/day shows the same reduction in
triglyceride levels as (-)-epicatechin (Dose: 0.1 mg/Kg/day) an
improvement of >30 fold.
Example 3: Activity of the Analogue of (+) Epicatechin of the
Present Invention
[0069] The compounds of the present invention were tested for their
activity on AMP kinase. The activity on AMP kinase was evaluated by
quantitative fluorescent immunoenzymatic assay of AMP kinase
phosphorylation status in cultured cells. The 5-AMP-activated
protein kinase (AMP kinase) is a key sensor of intracellular energy
balance. AMP kinase is activated in response to an increase in the
AMP/ATP ratio which can be caused by a number of factors such as
muscle contraction, starvation, or hypoxia. AMP kinase is a
heterotrimeric protein complex comprising of (63 kDa), -(38 kDa)
and - (38 kDa) subunits. For each subunit, isoforms have been
identified (1, 2, 1, 2, 1, 2, 3) which theoretically allow the
formation of 12 different proteins. The -subunit contains a
serine/threonine kinase domain and the regulatory subunits contain
binding sites for AMP and ATP (-subunit) and for glycogen
(-subunit). AMP kinase is activated by phosphorylation on Thr-172
within the catalytic domain. AMP binding results in a 2 to 5-fold
increase in AMP kinase activity compared to the basal level.
Binding of AMP to the -subunit causes allosteric activation of the
kinase and induces a conformational change in the kinase domain
that protects AMP kinase from dephosphorylation of Thr-172.
[0070] BioAssay Systems' cell-based ELISA measure phosphorylated
AMP kinase in whole cells and normalizes the signal to the total
protein content. The antibody recognizes both -subunits and, thus,
can be used for cells from all tissues (human, mouse, rat). This
simple and efficient assay eliminates the need for cell lysate
preparation and can be used to study AMP kinase regulation in
short-term and long-term assays. In this assay, cells grown in
96-well plates are fixed and permeabilized in the wells. AMP kinase
phosphorylation (pAMPK) is measured using a fluorescent ELISA
followed by total protein measurement in each well. Compound 1001,
exhibits AMPK activity at 1 nM.
Example 4: Determination of the Pharmacokinetic Parameters of the
Analogue of the Present Invention
[0071] Female Balb C mice 4 per group after overnight fasting were
dosed orally (via gavage) with compound 1 in 5% NMP in normal
saline (10 ml/kg). Blood was collected by serial bleeding at 0.16
hr, 0.5 hr, 1 hr, 2 hr, 4 hr, 6 hr, 8 h in heparinized tubes. Blood
samples were centrifuged at 10,000 rpm for 5 min. at 4.degree. C.
to obtain the plasma, which were aspirated into separate labeled
tubes and stored at -80.degree. C. 400 ng/ml of standard in
acetonitrile was used as the drug extraction solvent for extracting
drug from plasma. Extraction solvent was added to plasma was
vortexed and shaken on shaker for 10 minutes, centrifuged at 10,000
rpm for 10 minutes at 4.degree. C. Supernatant was kept for
analysis.
[0072] Acetonitrile and plasma calibration curves were generated
and percentage of drug recovery from plasma determined.
Quantitative analysis was done by liquid chromatography tandem mass
spectrometer (API3200 LC-MS/MS). C.sub.max, T.sub.max, AUC and ti/2
were calculated using Graph Pad PRISM version 5.04 and the results
were depicted in Table 3.
TABLE-US-00003 TABLE 3 Pharmacokinetic parameters of the compounds
of the present invention. PK STUDY (Oral) Elimination Dose Compound
AUC(nM*h) t1/2 (hr) (mpk) (+) Epicatechin 683 2.13 10 1001 2795.70
4.50 10
[0073] It may be noted that the compounds of the present invention
(1001) are suitable for administration.
* * * * *