Quinazolin-4(3H)-ones capable of upregulating the expression of endogenous apolipoprotein A-1

Article abstract of DOI:10.1517/13543776.2011.548322

An application claims non-naturally occurring upregulators of apolipoprotein A-I (ApoA-l) quinazolin-4(3H)-ones. The claimed quinazolin-4(3H)-ones are efficacious in upregulating the expression of endogeneous ApoA-l, and can potentially treat and prevent

Full text of DOI:10.1517/13543776.2011.548322

Patent Evaluation
Quinazolin-4(3H)-ones capable of
upregulating the expression of
endogenous apolipoprotein A-1
Jianhui Wu, Ming Zhao & Shiqi Peng†
1. Introduction
Capital Medical University, College of Pharmaceutical Sciences, Beijing 100069, PR China
2. Synthetic chemistry
3. Bio-assay and efficacy
4. Expert opinion
An application claims non-naturally occurring upregulators of apolipoprotein
A-I (ApoA-l) quinazolin-4(3H)-ones. The claimed quinazolin-4(3H)-ones are
efficacious in upregulating the expression of endogeneous ApoA-l, and can
potentially treat and prevent cardiovascular disease and related disease
states, including cholesterol or lipid-related disorders, such as atherosclerosis.
This application of Resverlogix Corp. increases the diversity of quinazolin-4
(3H)-ones capable of upregulating the expression of endogeneous ApoA-l.
Keywords: anti-atherogenic effect, apolipoprotein A-I, cardiovascular disease,
quinazolin-4(3H)-one, upregulators
Expert Opin. Ther. Patents (2011) 21(3):431-435
1. Introduction
Atherosclerotic coronary heart disease (CHD) is a major cause of death all over the
world and has been characterized by the elevated levels of cholesterol in circulating
plasma and the accumulated lipid in arterial lesion sites [1]. With epidemiologic
studies, the high level of low-density lipoprotein cholesterol and the low level of
high-density lipoprotein cholesterol (HDL-C) are correlated with the increased
risk of CHD [2,3]. The therapeutic intervention of CHD aims at either lowering
the level of cholesterol or raising the level of HDL-C [4-8]. Reducing the level of cho-
lesterol in the plasma may benefit 30% of individuals, while each 1 mg/dl increment
in serum HDL-C may lead to a 2% decrement in CHD risk due to the anti-
atherogeric effect on the recruitment of cholesterol from peripheral tissues and the
transportation of cholesterol towards the liver [9]. Apolipoprotein A-I (ApoA-I) is
a major form of HDL-C and has a major contribution in offering cardioprotective
action. Upregulating the expression of endogenous ApoA-I and consequently
increasing the circulation HDL-C are of clinical importance. Recombinant
ApoA-I and the peptide mimic of ApoA-I are currently available ApoA-I deriva-
tives [10], while polyphenols [11,12], flavanois [13,14], isoflavanoids [11,13], 4H-chro-
men-4-ones [13] and quinazolin-4(3H)-ones [15,16] synthesized by Resverlogix
Corp. are reported to be able to upregulate the expression of endogenous ApoA-I.
Comparing with ApoA-I derivatives lacking stability, these synthetic small mole-
cules are in an advantageous position. Among these small molecules, the structural
diversity of quinazolin-4(3H)-ones is greatly increased by this application.
Besides being the building block of ~ 150 natural alkaloids and clinic drugs [17],
quinazolin-4(3H)-ones possess a variety of pharmacological functions such as
anti-hyperlipidemic [17], antitumor [18], antimicrobial [19], antiviral [20], anti-
hypertensive [21], anti-inflammatory [22], anti-convulsant [23], anti-malarial [24],
anti-tuberculosis [25], anti-HIV [26], anti-ulcer [27], analgesic [28], inhibiting tyro-
sine kinase [29], inhibiting adenosine [30] and especially upregulating the expression
of endogenous ApoA-I explored by this application.
10.1517/13543776.2011.548322 © 2011 Informa UK, Ltd. ISSN 1354-3776
All rights reserved: reproduction in whole or in part not permitted
431

Quinazolin-4(3H)-ones capable of upregulating the expression of endogenous ApoA-1
R3
H
N
NH2
R1
R1
N
OH
O
O
R2
2-Aminobenzoic acid
Quinazolin-4(3H)-one
Triphosgene,
pyridine, ¦¤
R3
¦¤
O
R2
H
H
N
O
NH2
H2N
R1
R1
H
N
O
DMF, ¦¤
O
O
R2
Isatoic anhydride
2-Amino-N-(substituted-
phenyl)-benzamide
Scheme 1. General procedures for preparing substituted-quinazolin-4(3H)-ones.
dimethylsilane gave 2-{4-[2-(tert-bu-tyldimethylsilanyloxy)-
ethoxy]-3,5-dimethylphenyl}-3-(4-sec-butylphenyl)-3H-pyri-do
[4,3-d]pyrimidin-4-one (7). Removing of tert-butyldimethylsilane
from 7 provided 8.
2. Synthetic chemistry
Most of the claimed compounds are the substituted quinazo-
lin-4(3H)-ones and the preparation can be explained with the
procedures depicted in Scheme 1. In brief, in the presence of
triphosgene and pyridine, the dehydration of substituted-
2-amino-benzoic acid gave substituted isatoic anhydride.
The amidation of substituted isatoic anhydride with 4-
substituted-aniline provided 2-amino-N-(4-substituted-phenyl)-
benzamide. The condensation of 2-aminobenzamide and
substituted benzaldehyde gene rated quinazolin-4(3H)-ones.
A substantially same synthetic approach was used for
a specific claim of 3-(4-sec-butylphenyl)-2-[4-(2-hydro-
xyethoxy)-3,5-dimethyl]pyrido[4,3-d]pyridine-4(3H)-one (8),
and its preparation consisted of seven-step reactions (Scheme 2).
After the acetylation of 1, the formed 4-acetoxy-3,5-dimethylben-
zoic acid (2) was treated with oxalyl chloride to provide acetic
acid 4-chlorocarbonyl-2,6-dimethylphenyl ester (3). The cycliza-
tion of acetic acid ester 3 with 4-aminonicotinic acid generated
2,6- dimethyl-4-{4-oxo-4H-pyrido[4,3-d][1,3]oxazin-2-yl}phenyl
ester (4). The amidation of phenyl ester 4 with 4-(sec-butyl)-
aniline led the cyclization and the formation of 4-{3-(4-sec-
butylphenyl)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-2-yl}-2,
6-dime- thylphenyl ester (5). On the removal of acetyl 5, it
was converted into 3-(4-sec-butylphe-nyl)-2-(4-hydroxy-3,
5-dimehylphenyl)-3H-pyrido[4,3-d]-pyrimidin-4-one (6).
3. Bio-assay and efficacy
The efficacy of upregulating ApoA-I expression of quinazolin-4
(3H)-ones was measured with quantification of ApoA-I mRNA
assay. In the assay, ApoA-I mRNA in tissue cells and the tran-
scriptional upregulation of ApoA-I induced by quinazolin-4
(3H)-ones were quantitated. In a 24-well plate containing
400 µl of MEM supplemented with 0.5% FBS, ~ 2 Â 105
human HepG2 hepatoma cells were cultured at 37ꢀC and in
5% CO2. After 24 h incubation, the cells were treated with qui-
nazolin-4(3H)-ones. At harvesting time, the cultured media
were collected for ApoA-I and albumin ELISAs, the attached
cells were rinsed with 200 µl of PBS, treated with 85 µl of
cell lysis solution and incubated for 5 -- 10 min at room tem-
perature to complete the lysis and the detachment of the cells.
After the last wash, the wash buffer was completely aspirated
without allowing the wells to dry, 80 µl of E3 was added,
and the mRNA catcher PLUS plate was incubated at 68ꢀC
for 5 min to elute mRNA. The mRNA was isolated for one-
step real-time room temperature-PCR reaction. Using CT val-
ues, the real-time PCR data were analyzed. The results suggest
that when the concentration is £ 100 µM, 90 quinazolin-4
O-alkylation of
6
with (2-bromoethoxyl)-tert-butyl-
432
Expert Opin. Ther. Patents (2011) 21(3)

Wu, Zhao & Peng
Expert Opin. Ther. Patents (2011) 21(3)
433

Quinazolin-4(3H)-ones capable of upregulating the expression of endogenous ApoA-1
(3H)-ones are able to effectively increase the expression of
endohereous ApoA-I by at least 20%.
uncertain, but the importance of their mechanisms of action,
the level of efficacy and the ripe preparation procedure of
quinazolin-4(3H)-ones make the investigators confident in
selecting the candidates for the development.
4. Expert opinion
Patent details
Resverlogix Corp. has continually made progress in the
discovery of synthetic small molecules, including quinazo-
lin-4(3H)-ones in particular, capable of upregulating the
expression of endohereous ApoA-I. This application with
the previous one (WO 2008/09223 A1) together presented
128 novel quinazolin-4(3H)-ones. In addition to the
biological data evidencing their use in increasing the level
of endohereous ApoA-I and consequently in treating
atherosclerotic CHD, in this application the synthetic
approaches to the construction of diverse quinazolin-4
(3H)-ones were well documented. These approaches were
generally characterized by the dehydration of substituted-
2-aminobenzoic acid, the amidation of substituted isatoic
anhydride with 4-substitutedaniline and the condensation
of 2-ami- no-N-(4-substitutedphenyl)-benzamide.
Title: Compound for the prevention and treatment of
cardiovascular disease
Assignee: Resverlogix Corp.
Inventors: Hansen C, Henrik
Priority date: 08/01/2009
Filing date: 08/01/2010
Publication date: 15/07/2010
Publication no: WO2010079431
Declaration of interest
J WU is supported by a grant from the National
Scientific Foundation of China (30901843) and the Beijing
Municipal Commission of Education (KM20091002010).
S Peng is supported by a grant from PHR (1HLB:
KZ200810025010).
Whether any quinazolin-4(3H)-one of the 128 novel
quinazolin-4(3H)-ones could progress to clinical trials is
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