Synthesis and biological activity of fibrate-based acyl- and alkyl-phenoxyacetic methyl esters and 1,2-dihydroquinolines

MEDICINAL

CHEMISTRY

RESEARCH

Medicinal Chemistry Research

https://doi.org/10.1007/s00044-019-02496-1

ORIGINAL RESEARCH

Synthesis and biological activity of ﬁbrate-based acyl- and alkyl-

phenoxyacetic methyl esters and 1,2-dihydroquinolines

Abraham Pucheta^1,2Aarón Mendieta^1,3Damián A. Madrigal^1,2Roberto I. Hernández-Benitez¹Liseth Romero²

●

Leticia Garduño-Siciliano²Catalina Rugerio-Escalona³María C. Cruz-López³Fabiola Jiménez³

●

Alejandra Ramírez-Villalva⁴Aydeé Fuentes-Benites⁴Carlos González-Romero⁴Omar Gómez-García¹

●

Julio López⁵Miguel A. Vázquez⁵Blanca Rosales-Acosta⁶Lourdes Villa-Tanaca⁶Alfonso Sequeda-Juárez⁷

●

1

Eva Ramón-Gallegos⁷Germán Chamorro-Cevallos²Francisco Delgado¹Joaquín Tamariz

●

Received: 10 October 2019 / Accepted: 22 December 2019

Abstract

A series of highly potent antihyperlipidemic agents constituted by a ﬁbrate-based structure was recently reported by our

group, whose synthesis started from isovanillin derivatives. In the interest of evaluating the bioisosteric effect of the vanillin-

based isomers on their antihyperlipidemic activity, the present study focuses on the synthesis of 5-acyl-1-phenoxyacetic

methyl esters 5a–c and their saturated side-chain 5-alkyl-1-phenoxyacetates 6a–c. Their strong in vivo effect was associated

with the inhibition of HMG-CoA reductase. Since 1,2-dihydroquinolines inhibit this enzyme, a series of such heterocycles

(9a–d) was prepared by our efﬁcient regioselective, one-step, solvent-free method. Apart from showing hypolipidemic

activity in vivo, some of the compounds displayed antifungal, antioxidant and cytotoxic activity in vitro. The binding mode

of four compounds at the active site of HMGRh was examined with docking simulations, observing an interaction with most

of the amino acids targeted by simvastatin.

●

Keywords Antihyperlipidemic activity 2-Acyl-1-hydroxyphenoxyacetic esters 2-Alkyl-1-hydroxyphenoxyacetic esters

●

1,2-Dihydroquinolines HMG-CoA reductase Antioxidant activity

Introduction

Hyperlipidemia, which represent a signiﬁcant cardiovas-

cular risk factor, is a growing public health problem

worldwide (Thagizadeh et al. 2019). It is mainly associated

with diseases such as atherosclerosis (Yuan et al. 2007;

Steinberg 2005) and ischemic heart disease (Mehta et al.

Supplementary information The online version of this article (https://

doi.org/10.1007/s00044-019-02496-1) contains supplementary

material, which is available to authorized users.

* Germán Chamorro-Cevallos

4

Departamento de Química Orgánica, Facultad de Química,

Universidad Autónoma del Estado de México (UAEM), Paseo

Tollocan y Colón S/N, 50000 Toluca, Mexico

gchamcev@yahoo.com.mx

* Joaquín Tamariz

jtamarizm@gmail.com

5

Departamento de Química, Universidad de Guanajuato, Noria Alta

S/N, 36050 Guanajuato, Gto, Mexico

1

Departamento de Química Orgánica, Escuela Nacional de Ciencias

Biológicas, Instituto Politécnico Nacional (IPN). Prol. Carpio y

Plan de Ayala S/N, 11340 Mexico City, Mexico

6

Laboratorio de Genética Microbiana, Escuela Nacional de

Ciencias Biológicas, Instituto Politécnico Nacional, Prol. Carpio y

Plan de Ayala S/N, 11340 Mexico City, Mexico

2

Departamento de Farmacia, Escuela Nacional de Ciencias

Biológicas, Instituto Politécnico Nacional, Av. Wilfrido Massieu

S/N, Unidad Adolfo López Mateos, 07738 Mexico City, Mexico

7

Departamento de Morfología, Escuela Nacional de Ciencias

Biológicas, Instituto Politécnico Nacional, Av. Wilfrido Massieu

S/N, Unidad Adolfo López Mateos, 07738 Mexico City, Mexico

3

Centro de Investigación en Biotecnología Aplicada, IPN. Carretera

Estatal Santa Inés Tecuexcomac-Tepetitla, Km 1.5, Tepetitla de

Lardizábal, 90700 Tlaxcala, Mexico

Medicinal Chemistry Research

2014). Obesity and diabetes may contribute substantially to

the elevated prevalence of dyslipidemia (Van Gaal et al.

2006; Aguilar-Salinas et al. 2001). When lifestyle changes

(e.g., diet, exercise, and weight loss) fail to control dysli-

pidemia, healthcare systems turn to the prescription of lipid-

lowering medication (Gielen et al. 2009; Grundy et al.

2005). Although many synthetic drugs are available to treat

hyperlipidemia, they are related with multiple adverse

effects, such as myopathy, rhabdomyolysis, and liver toxi-

city (Chalasani 2005; Graham et al. 2004; Elisaf et al. 2008;

Hodel 2002; Joy and Hegele 2009; Williams and Feely

2002; Davidson et al. 2007).

Statins are among the most commonly used drugs for the

clinical management of hypercholesterolemia (Maron et al.

2000; Wierzbicki 2001; Miyazaki et al. 2004; Vivancos

et al. 1999). Their adverse effects include rhabdomyolysis,

cognitive decline, neuropathy, pancreatic, and hepatic dys-

function, and an increased risk of cancer (Golomb and Evans

2008; Rotta-Bonﬁn et al. 2015). The mechanism of their

activity has been established as the inhibition of the

3-hydroxy-3-methylglutaryl-coenzyme

A

(HMG-CoA)

reductase (HMGR), an enzyme involved in the biosynthesis

of cholesterol (Singh et al. 2009; Haines et al. 2013).

Among other agents inhibiting HMGR (Menéndez et al.

2001; Liu and Yeh 2002; Parker et al. 1993; Sung et al.

2004; Bradfute and Simoni 1994; Harada-Shiba et al. 1995;

Panini et al. 1986) are cholestin (Man et al. 2002), dios-

genin (Raju and Bird 2007), lanosterol analogs (Trzaskos

et al. 1993), β-sitosterol (Field et al. 1997), and the natural

product α-asarone (1) (active metabolite of the Yucatan

peninsula (Mexico) tree called Elemuy (Mosannona

depressa (Baill.) Chatrou) (Campos-Ríos and Chiang Cab-

rera 2006)) resulting in an antihyperlipidemic effect (Cha-

morro et al. 1993). α-Asarone (1) has also been reported to

inhibit HMGR (Singh et al. 2009; Rodríguez-Páez et al.

2003) and exhibit antifungal (Momin and Nair 2002; Lee

et al. 2004) and antithrombotic (Poplawsky et al. 2000)

activity.

Fibrates such as cloﬁbrate (2a) (Oliver 2012; Moham-

madzadeh et al. 2013), bezaﬁbrate (2b) (BIP study group

2000), and fenoﬁbrate (2c) (Uchida et al. 2011) are

phenoxyacetic-based compounds with a potent hypo-

triglyceridemic activity (Lalloyer and Staels 2010; Jover-

Fernández and Hernández-Mijares 2012). Unlike statins,

their mechanism of action consists of binding to and acti-

Fig. 1 Structures of α-asarone (1), cloﬁbrate (2a), bezaﬁbrate (2b),

fenoﬁbrate (2c), α-asarone analogs 3a–f, and ﬁbrate analogs 4a–f

cloﬁbrate was withdrawn from the market in some countries.

The biological activity of phenoxy acids and their derivatives

is exceptionally wide-ranging, being antihyperlipidemic,

hypoglycemic, antimicrobial, antiviral, antitubercular, anti-

inﬂammatory, analgesic, antioxidant, anticancer, and anti-

hypertensive (Shaheen et al. 2016).

In the search of new synthetic hypolipidemic agents with

fewer or no side effects, our group previously synthesized a

series of α-asarone (1) analogs that showed the desired activity

in potent form (Argüelles et al. 2010; Cruz et al. 2003).

Docking studies revealed the binding mode of 1 with the

active site of the human HMGR (HMGRh) enzyme (Medina-

Franco et al. 2005). Moreover, the oxyacetic group at the C-2

position of 1 (Argüelles et al. 2010), mimicking the ﬁbrate

structure, proved to have a signiﬁcant lipid-lowering activity

(Labarrios et al. 1999; Zúñiga et al. 2005). Particularly, the

series of α-asarone- 3a–f and phenoxyacetic-based derivatives

4a–f (Fig. 1) (all prepared from isovanillin) displayed a sharp

reduction in cholesterol, LDL-cholesterol, and triglycerides,

indicating HMGRh inhibition (Mendieta et al. 2014).

vating peroxisome proliferator activated receptor

α

(PPARα), a transcription factor (Schoonjans et al. 1996;

Willson et al. 2000; Fazio and Linton 2004; Shaikh and Ali

2018). Fibrates enhance HDL cholesterol but can cause

myopathy, cholelithiasis, nausea, syndrome of inappropriate

secretion of antidiuretic hormone ADH (SIADH), and liver

injury (The Field Study 2005; Okopień et al. 2018). These

adverse side effects were among other motives that

On the other hand, there is evidence of diverse types of

biological activity of 1,2-dihydroquinolines, found to act as

antidiabetic, anti-inﬂammatory (Hegedüs et al. 2007), anti-

bacterial (Johnson et al. 1989), antioxidant and cytotoxic

(Dorey et al. 2000; Błaszczyk and Skolimowski 2007;

Błaszczyk et al. 2013; de Koning 2002) (Błaszczyk et al.

Medicinal Chemistry Research

Materials and methods

Chemistry

Melting points were determined with an Electrothermal

capillary melting point apparatus. IR spectra were recorded

on an FT-IR Perkin Elmer 2000 spectrophotometer. ¹H (300

or 500 MHz) and ¹³C (75.4 or 125 MHz) NMR spectra were

recorded on Varian Mercury-300 or Varian VNMR System

instruments, with TMS as internal standard. Mass spectra

(MS) were taken in the electron impact (EI) mode on

Hewlett-Packard 5971A and Thermo-Finnigan Polaris Q

spectrometers. High-resolution mass spectra (HRMS) were

obtained (EI) on a Jeol JSM-GCMateII spectrometer.

Commercial reagents were used as received from Aldrich

and anhydrous solvents were obtained by a distillation

process. Thin layer chromatography was performed on

precoated silica gel plates (Merck 60F₂₅₄). Silica gel

(230–400 mesh) was used for column chromatography. All

air moisture sensitive reactions were carried out with oven-

dried glassware under nitrogen atmosphere. Prior to use,

acetone and DMF were distilled from 4Å; molecular sieves

and methylene chloride from calcium hydride. K₂CO₃was

dried overnight at 120 °C before use. All other reagents

were used without further puriﬁcation. The preparation of

compounds 12 (Grenier et al. 2000; Kiss et al. 2010; Rashid

et al. 2018) and 9a–d has been previously described

(Gutiérrez et al. 2013).

Fig. 2 Structures of the synthesized ﬁbrate-based analogs 5a–c, 6a–c,

and 7a–c, and the 1,2-dihydroquinolines 9a–d

2006; Błaszczyk and Skolimowski 2005; Błaszczyk and

Skolimowski 2006) agents. Likewise, analogous 1,2,3,4-

tetrahydroquinolines have shown antifungal (Chander et al.

2016a), antimicrobial (Chander et al. 2016b), and anti-

oxidant (Dorey et al. 2000) activity, which has been asso-

ciated with their capacity of lipid peroxidation and the

inhibition of HMGR (Hegedüs et al. 2007). Consequently,

these heterocycles may also produce a general hypolipi-

demic, antioxidant and antifungal effect.

The ﬁrst aim of the present study was to examine the

possible impact on the antihyperlipidemic activity caused

by the isomeric position of the substituents in the benzene

ring of vanillin-based isomers. Thus, three series of ﬁbrate-

based compounds were synthesized starting from vanillin

(8) and their structure elucidated. The series consisted of 5-

acyl-1-phenoxyacetic methyl esters 5a–c, their saturated

side-chain 5-alkyl-1-phenoxyacetates 6a–c and nitrophe-

noxyacetates 7a–c (Fig. 2). Due to the HMGR-related

activity of hydroquinolines, a series of the latter was syn-

thesized by using our regioselective one-step method

(Gutiérrez et al. 2013). These compounds, 9a–d, were

evaluated in vivo as hypolipidemic agents, and in vitro for

their antifungal, antioxidant, and anticancer effect. The

mechanism of action of the test compounds on the HMGRh

structure was explored with docking simulations.

Methyl 2-(4-formyl-2-methoxyphenoxy)acetate (10)

Under an N₂atmosphere, a mixture of vanillin (8) (10.00 g,

65.72 mmol), methyl bromoacetate (16.463 g, 98.58 mmol),

and dry K₂CO₃(18.165 g, 131.44 mmol) in anhydrous

acetone (100 mL) was heated to 60 °C for 4 h. The reaction

mixture was ﬁltered and the solvent removed under

vacuum. The residue was puriﬁed by column chromato-

graphy over silica gel (20 g/g of crude, hexane/EtOAc, 8:2)

to give 10 (13.41 g, 91%) as a white solid. Rf 0.48 (hexane/

EtOAc, 6:4); m.p. 92–93 °C. IR (KBr): ν 2994, 1740, 1677,

1591, 1508, 1452, 1404, 1285, 1261, 1224, 1136, 1028,

1

861, 807, 735 cm⁻¹. H NMR (500 MHz, CDCl₃): δ 3.81

(s, 3H, CO₂CH₃), 3.96 (s, 3H, OCH₃), 4.80 (s, 2H,

CH₂CO₂CH₃), 6.89 (d, J = 8.5 Hz, 1H, H-6′), 7.42 (dd, J =

8.5, 1.8 Hz, 1H, H-5′), 7.44 (d, J = 1.8 Hz, 1H, H-3′), 9.87

(s, 1H, CHO). ¹³C NMR (125 MHz, CDCl₃): δ 52.4

(CO₂CH₃), 56.0 (OCH₃), 65.8 (CH₂CO₂CH₃), 109.8 (C-3′),

112.3 (C-6′), 126.1 (C-5′), 131.2 (C-4′), 149.9 (C-2′), 152.4

(C-1′), 168.5 (CO₂CH₃), 190.7 (CHO). MS (70 eV): m/z

224 (M⁺, 100), 165 (25), 150 (30), 137 (35), 119 (36), 105

(20), 95 (30), 77 (31), 59 (32). HRMS (EI): calcd. for

C₁₁H₁₂O₅[M]⁺224.0685; found: 224.0685.

Medicinal Chemistry Research

Methyl 2-(4-hydroxy-2-methoxyphenoxy)acetate

(11)

OCH₃), 4.62 (s, 2H, CH₂CO₂CH₃), 6.46 (s, 1H, H-3′), 7.29

(s, 1H, H-6′), 12.61 (s, 1H, OH). ¹³C NMR (125 MHz,

CDCl₃): δ 26.3 (CH₃CO), 52.2 (CO₂CH₃), 56.1 (OCH₃),

68.1 (CH₂CO₂CH₃), 100.9 (C-3′), 111.9 (C-5′), 118.3 (C-

6′), 139.6 (C-1′), 157.8 (C-2′), 161.2 (C-4′), 169.5

(CO₂CH₃), 202.2 (CH₃CO). MS (70 eV): m/z 254 (M⁺, 51),

239 (6), 195 (7), 181 (100), 153 (14), 135 (15), 107 (9).

HRMS (EI): calcd for C₁₂H₁₄O₆[M]⁺: 254.0790; found:

254.0787.

A mixture of 10 (10,000 g, 44.64 mmol) and mCPBA (77%)

(15,000 g, 66.96 mmol) in CH₂Cl₂(100 mL) was stirred at

room temperature (rt) for 12 h. The solvent was removed

under vacuum, and MeOH (100 mL) and HCl (6N) (5 mL)

were added. The reaction mixture was stirred at rt for 1 h

before removing the solvent under vacuum. A saturated

aqueous solution of NaHCO₃was added until neutral, then

extracted with EtOAc (4 × 50 mL). The organic layer was

dried (Na₂SO₄) and the solvent removed under vacuum. The

residue was puriﬁed by column chromatography over silica

gel (20 g/g of crude, hexane/EtOAc, 7:3) to afford 11

(6.814 g, 72%) as a white solid. Rf 0.43 (hexane/EtOAc,

6:4); m.p. 89–90 °C. IR (KBr): ν 3431, 2958, 1732, 1612,

1512, 1479, 1433, 1309, 1208, 1136, 1081, 1032, 953, 837,

793, 766, 712 cm⁻¹. ¹H NMR (300 MHz, CDCl₃): δ 3.79 (s,

3H, CO₂CH₃), 3.80 (s, 3H, OCH₃), 4.62 (s, 2H,

CH₂CO₂CH₃), 4.96 (br, 1H, OH), 6.30 (dd, J = 8.5, 2.7 Hz,

1H, H-5′), 6.46 (d, J = 2.7 Hz, 1H, H-3′), 6.75 (d, J =

8.5 Hz, 1H, H-6′). ¹³C NMR (75 MHz, CDCl₃): δ 52.2

(CO₂CH₃), 55.7 (OCH₃), 67.6 (CH₂CO₂CH₃), 100.7 (C-3′),

106.0 (C-5′), 116.7 (C-6′), 140.9 (C-1′), 150.7 (C-2′), 151.8

(C-4′), 170.2 (CO₂CH₃). HRMS (EI): calcd. for C₁₀H₁₂O₅

[M]⁺212.0685; found: 212.0681.

Methyl 2-(4-hydroxy-2-methoxy-5-

propionylphenoxy)acetate (5b)

Following the general procedure, 11 (1.000 g, 4.72 mmol)

was mixed with propanoyl chloride (0.654 g, 7.07 mmol)

and BF₃·OEt₂(0.670 g, 4.72 mmol) to obtain 5b (0.91 g,

76%) as a white solid. Rf 0.40 (hexane/EtOAc, 7:3); m.p.

105–106 °C. IR (KBr): ν 3482, 2979, 2942, 1748, 1638,

1616, 1565, 1504, 1439, 1375, 1319, 1249, 1193, 1158,

1

1069, 1007, 975, 945, 820, 707 cm⁻¹. H NMR (300 MHz,

CDCl₃): δ 1.22 (t, J = 7.2 Hz, 3H, CH₃CH₂CO), 2.90 (q,

J = 7.2 Hz, 2H, CH₃CH₂CO), 3.80 (s, 3H, CO₂CH₃), 3.90

(s, 3H, OCH₃), 4.62 (s, 2H, CH₂CO₂CH₃), 6.46 (s, 1H, H-

3′), 7.33 (s, 1H, H-6′), 12.70 (s, 1H, OH). ¹³C NMR

(75 MHz, CDCl₃): δ 8.3 (CH₃CH₂CO), 31.2 (CH₃CH₂CO),

52.1 (CO₂CH₃), 56.0 (OCH₃), 68.0 (CH₂CO₂CH₃), 100.9

(C-3′), 111.3 (C-5′), 117.5 (C-6′), 139.5 (C-1′), 157.4 (C-

2′), 161.1 (C-4′), 169.5 (CO₂CH₃), 204.9 (CH₃CH₂CO).

MS (70 eV): m/z 268 (M⁺, 66), 239 (66), 195 (100), 138

(22), 121 (14). HRMS (EI): m/z calcd for C₁₃H₁₆O₆[M]⁺:

268.0947; found: 268.0946.

General procedure for preparing 5-

acylphenoxyacetic methyl esters 5a–c

Under an N₂atmosphere and at rt, a mixture of 11 (1.0 mol

equiv.) and of the respective acyl chloride (1.5 mol equiv.)

was stirred for 10 min. Subsequently, BF₃·OEt₂(1.0 mol

equiv.) was added, stirred at rt for 20 min, and then heated

to 80 °C for 4 h. Ice was poured into the reaction mixture,

followed by adding a saturated aqueous solution of

NaHCO₃dropwise until neutral, then washed with EtOAc

(3 × 50 mL). The organic extract was dried (Na₂SO₄) and

the solvent removed under vacuum. The residue was pur-

iﬁed by column chromatography over silica gel (20 g/g of

crude, hexane/EtOAc, 85:15).

Methyl 2-(5-butyryl-4-hydroxy-2-methoxyphenoxy)

acetate (5c)

Following the general procedure, 11 (1.000 g, 4.72 mmol)

was mixed with butanoyl chloride (0.753 g, 7.07 mmol) and

BF₃·OEt₂(0.670 g, 4.72 mmol) to produce 5c (0.892 g,

67%) as a white solid. Rf 0.46 (hexane/EtOAc, 7:3); m.p.

82–83 °C. IR (KBr): ν 3482, 2954, 1748, 1637, 1511, 1442,

1384, 1336, 1283, 1253, 1197, 1162, 1075, 1019, 986, 900,

848, 793, 703 cm⁻¹

.

¹H NMR (300 MHz, CDCl₃):

Methyl 2-(5-acetyl-4-hydroxy-2-methoxyphenoxy)

acetate (5a)

δ 1.01 (t, J = 7.5 Hz, 3H, CH₃(CH₂)₂CO), 1.75 (sext, J =

7.5 Hz, 2H, CH₃CH₂CH₂CO), 2.84 (t, J = 7.5 Hz, 2H,

CH₃CH₂CH₂CO), 3.81 (s, 3H, CO₂CH₃), 3.91 (s, 3H,

OCH₃), 4.63 (s, 2H, CH₂CO₂CH₃), 6.46 (s, 1H, H-3′), 7.32

(s, 1H, H-6′), 12.78 (s, 1H, OH). ¹³C NMR (75 MHz,

CDCl₃): δ 13.8 (CH₃(CH₂)₂CO), 18.1 (CH₃CH₂CH₂CO),

39.9 (CH₃CH₂CH₂CO), 52.2 (CO₂CH₃), 56.1 (OCH₃), 68.0

(CH₂CO₂CH₃), 100.87 (C-3′), 111.4 (C-5′), 117.5 (C-6′),

139.5 (C-1′), 157.4 (C-2′), 161.2 (C-4′), 169.5 (CO₂CH₃),

204.6 (CH₃(CH₂)₂CO). MS (70 eV): m/z 282 (M⁺, 66), 239

(68), 209 (100), 167 (14), 138 (20), 107 (10), 69 (16).

Following the general procedure, 11 (1.000 g, 4.72 mmol)

was mixed with acetyl chloride (0.555 g, 7.07 mmol) and

BF₃·OEt₂(0.670 g, 4.72 mmol) to provide 5a (0.91 g, 76%)

as a white solid. Rf 0.31 (hexane/EtOAc, 7:3); m.p.

125–126C. IR (KBr): ν 3472, 2974, 2372, 1744, 1640,

1614, 1503, 1440, 1365, 1334, 1265, 1202, 1163, 1058,

1

1007, 975, 814, 710 cm⁻¹. H NMR (500 MHz, CDCl₃):

δ 2.53 (s, 3H, CH₃CO), 3.80 (s, 3H, CO₂CH₃), 3.90 (s, 3H,

Medicinal Chemistry Research

HRMS (EI): m/z calcd for C₁₄H₁₈O₆[M]⁺: 282.1103;

found: 282.1100.

67.8 (CH₂CO₂CH₃), 100.9 (C-3′), 118.1 (C-6′), 119.5 (C-

5′), 140.5 (C-1′), 148.5 (C-2′), 149.1 (C-4′), 170.3

(CO₂CH₃). HRMS (EI): m/z calcd for C₁₃H₁₈O₅[M]⁺:

254.1154; found: 254.1151.

General procedure for preparing 5-

alkylphenoxyacetic methyl esters 6a–c

Methyl 2-(5-butyl-4-hydroxy-2-methoxyphenoxy)acetate

(6c)

Under an N₂atmosphere and at rt, a mixture of Zn/Hg and

HCl (36%) (1.0 g/1.0 mL, 10.0 mol equiv.) was stirred for

5 min before being poured into a ﬂask. Addition was made

of the corresponding acyl derivative 5a–c (1.0 mol equiv.),

followed by a mixture of MeOH/HCl (1.0/0.2 mL). After

heating to 60 °C for 4 h, ice was poured in and a saturated

aqueous solution of NaHCO₃added dropwise until neutral,

and then washed with EtOAc (3 × 30 mL). The organic

extract was dried (Na₂SO₄) and the solvent was removed

under vacuum. The residue was puriﬁed by column chro-

matography over silica gel (20 g/g of crude, hexane/

EtOAc, 95:5).

Following the general procedure, 5c (1.000 g, 3.55 mmol)

was mixed with Zn/Hg (9.43 g, 35.5 mmol), resulting in 6c

(0.65 g, 68%) as a white solid. Rf 0.31 (hexane/EtOAc, 7:3);

m.p. 49–50 °C. IR (KBr): ν 3448, 3019, 2952, 2860, 1719,

1617, 1529, 1449, 1417, 1367, 1306, 1206, 1123, 1078,

1013, 939, 894, 856, 829, 759, 714 cm⁻^{1 1}H NMR

.

(500 MHz, CDCl₃): δ 0.94 (t, J = 7.5 Hz, 3H, H-4″), 1.35

(sext, J = 7.5 Hz, 2H, H-3″), 1.53 (qu, J = 7.5 Hz, 2H, H-

2″), 2.45 (t, J = 7.5 Hz, 2H, H-1″), 3.79 (s, 3H, CO₂CH₃),

3.80 (s, OCH₃), 4.61 (s, 2H, CH₂CO₂CH₃), 6.42 (s, 1H, H-

3′), 6.70 (s, 1H, H-6′). ¹³C NMR (125 MHz, CDCl₃): δ 13.9

(C-4″), 22.4 (C-3″), 29.0 (C-1″), 32.1 (C-2″), 52.1

(CO₂CH₃), 56.0 (OCH₃), 68.0 (CH₂CO₂CH₃), 101.1 (C-3′),

118.4 (C-6′), 119.4 (C-5′), 141.0 (C-1′), 148.9 (C-2′, C-4′),

170.0 (CO₂CH₃). HRMS (EI): m/z calcd for C₁₄H₂₀O₅

[M]⁺: 268.1311; found: 268.1303.

Methyl 2-(5-ethyl-4-hydroxy-2-methoxyphenoxy)acetate

(6a)

Following the general procedure, 5a (1.000 g, 3.94 mmol)

was mixed with Zn/Hg (10.47 g, 39.4 mmol) to furnish 6a

(0.68 g, 72%) as a white solid. Rf 0.26 (hexane/EtOAc, 7:3);

m.p. 66–67 °C. IR (KBr): ν 3402, 2961, 2925, 2866, 1750,

1717, 1616, 1525, 1447, 1423, 1367, 1293, 1237, 1204,

1122, 1069, 1007, 857, 747, 706 cm⁻¹. ¹H NMR (500 MHz,

CDCl₃): δ 1.17 (t, J = 7.5 Hz, 3H, H-2″), 2.52 (q, J =

7.5 Hz, 2H, H-1″), 3.78 (s, 6H, CO₂CH₃, OCH₃), 4.62 (s,

2H, CH₂CO₂CH₃), 4.77–4.84 (m, 1H, OH), 6.42 (s, 1H, H-

3′), 6.73 (s, 1H, H-6′). ¹³C NMR (125 MHz, CDCl₃): δ 14.2

(C-2″), 22.3 (C-1″), 52.1 (CO₂CH₃), 55.9 (OCH₃), 68.0

(CH₂CO₂CH₃), 101.1 (C-3′), 117.7 (C-6′), 120.9 (C-5′),

141.0 (C-1′), 148.8 (C-2′, C-4′), 170.1 (CO₂CH₃). HRMS

(EI): m/z calcd for C₁₂H₁₆O₅[M]⁺: 240.0998; found:

240.1000.

Methyl 2-(4-Formyl-2-methoxy-6-nitrophenoxy)acetato (7a)

Under N₂atmosphere,

a mixture of 12 (10.000 g,

50.76 mmol), dry K₂CO₃(14,010 g, 101.52 mmol) and

methyl bromoacetate (15.10 g, 98.7 mmol) in anhydrous

DMF (50 mL) was stirred at 60 °C for 4 h (Brown et al.

2010). The reaction mixture was ﬁltered, the solvent

removed under vacuum, and the residue puriﬁed by column

chromatography over silica gel (20 g/g of crude, hexane/

EtOAc, 8:2) to give 7a (9150 g, 67%) as a yellow solid. Rf

0.66 (hexane/EtOAc, 1:1); m.p. 96–97 °C. IR (KBr): ν

3385, 3086, 3014, 2956, 2851, 1752, 1701, 1604, 1541,

1466, 1427, 1357, 1289, 1218, 1202, 1141, 1046, 918, 872,

772, 680 cm⁻¹. ¹H NMR (300 MHz, CDCl₃): δ 3.78 (s, 3H,

CO₂CH₃), 3.99 (s, 3H, OCH₃), 4.90 (s, 2H, CH₂CO₂CH₃),

7.64 (d, J = 1.8 Hz, 1H, H-3′), 7.88 (d, J = 1.8 Hz, 1H,

H-5′), 9.93 (s, 1H, CHO). ¹³C NMR (75 MHz, CDCl₃):

δ 52.3 (CO₂CH₃), 56.8 (OCH₃), 69.4 (CH₂CO₂CH₃), 113.5

(C-3′), 119.7 (C-5′), 131.7 (C-4′), 145.2 (C-1′), 153.6

(C-2′), 168.5 (CO₂CH₃), 188.9 (CHO). HRMS (EI): m/z

calcd for C₁₁H₁₁NO₇[M]⁺: 269.0535; found: 269.0541.

Methyl 2-(4-hydroxy-2-methoxy-5-propylphenoxy)acetate

(6b)

Following the general procedure, 5b (1.000 g, 3.73 mmol)

was mixed with Zn/Hg (9.93 g, 37.3 mmol) to form 6b

(0.663 g, 70%) as a white solid. Rf 0.30 (hexane/EtOAc,

7:3); m.p. 54–55 °C. IR (KBr): ν 3390, 3019, 2955, 2867,

1871, 1708, 1619, 1528, 1450, 1419, 1377, 1302, 1198,

1

1120, 1078, 1008, 884, 866, 843, 768, 739, 703 cm⁻¹. H

NMR (300 MHz, CDCl₃): δ 0.93 (t, J = 7.4 Hz, 3H, H-3″),

1.57 (sext, J = 7.4 Hz, 2H, H-2″), 2.46 (t, J = 7.4 Hz, 2H,

H-1″), 3.73 (s, 3H, OCH₃), 3.78 (s, 3H, CO₂CH₃), 4.61 (s,

2H, CH₂CO₂CH₃), 5.45 (br s, 1H, OH), 6.42 (s, 1H, H-3′),

6.69 (s, 1H, H-6′). ¹³C NMR (75 MHz, CDCl₃): δ 13.8 (C-

3″), 23.0 (C-2″), 31.3 (C-1″), 52.1 (CO₂CH₃), 55.7 (OCH₃),

Methyl 2-(4-hydroxy-2-methoxy-6-nitrophenoxy)acetate

(7b)

A mixture of 7a (1.000 g, 3.72 mmol) and mCPBA (77%)

(1.240 g, 5.57 mmol) in CH₂Cl₂(10 mL) was stirred at rt for

4 h before removing the solvent under vacuum. MeOH

Medicinal Chemistry Research

(10 mL) and HCl (37%) (1 mL) were added, the mixture

was stirred at rt for 2 h, and the solvent was removed under

vacuum. An aqueous saturated solution of NaHCO₃was

added until neutral, then extracted with EtOAc (4 × 50 mL).

The organic layer was dried (Na₂SO₄), the solvent removed

under vacuum, and the residue puriﬁed by column chro-

matography over silica gel (20 g/g of crude, hexane/EtOAc,

1:1) to afford 7b (0.580 g, 61%) as a pale yellow solid. Rf

0.33 (EtOAc); m.p. 110–111 °C. IR (KBr): ν 3070, 2952,

2643, 1732, 1609, 1542, 1490, 1466, 1421, 1354, 1289,

1222, 1202, 1114, 1066, 1017, 918, 897, 865, 854, 814,

showed no gross abnormalities in any treated mice. All

animals were handled and maintained in accordance with

ethical principles and regulations speciﬁed by the Animal

Care and Use Committee of our institution and the stan-

dards of the National Institutes of Health of Mexico.

An aqueous solution of Triton WR 1339 was adminis-

tered intraperitoneally (ip) to mice (400 mg/kg) and after 1 h

the test compounds (25, 50, or 100 mg/kg, dissolved in

saline, or saline-Tween) were injected ip. Blood was taken

from a retro-orbital puncture 24 h later, and the plasma

levels of total cholesterol (TC) and triglycerides (TG) were

determined in duplicate with commercially available kits.

Values are expressed as the mean from 6 animals per

compound.

1

775, 755, 736, 700 cm⁻¹. H NMR (500 MHz, CDCl₃): δ

3.79 (s, 3H, CO₂CH₃), 3.99 (s, 3H, OCH₃), 4.89 (s, 2H,

CH₂CO₂CH₃), 7.80 (d, J = 2.0 Hz, 1H, H-3′), 8.12 (d, J =

2.0 Hz, 1H, H-5′). ¹³C NMR (125 MHz, CDCl₃): δ 52.3

(CO₂CH₃), 56.8 (OCH₃), 69.5 (CH₂CO₂CH₃), 116.8 (C-3′),

118.7 (C-5′), 124.6 (C-6′), 144.6 (C-1′), 153.0 (C-2′), 168.6

(CO₂CH₃), 169.1 (C-4′). HRMS (EI): m/z calcd for

C₁₀H₁₁NO₇[M]⁺: 257.0536; found: 257.0531.

Antifungal activity

The in vitro antimicrobial effect of compounds 5a–c, 6a–b,

7a, 7c, and 9a–d was evaluated with susceptibility assays

based on the microdilution techniques of the Antimicrobial

Susceptibility Testing Protocols standardized by the Clin-

ical and Laboratory Standards Institute (CLSI). The M38-

A2 method was used for the four strains of ﬁlamentous

fungi (CLSI 2002; Espinel-Ingroff and Canton 2007a):

Aspergillus fumigatus ATCC-16907, Trichosporon cuta-

neum ATCC-28592, Rhizopus oryzae ATCC-10329 and

Mucor hiemalis ATCC-8690. The M27-A3 method was

applied for the three strains of ATCC Candida yeasts (CLSI

2008; Espinel-Ingroff and Cantón 2007b; Pfaller and Die-

kema 2012): C. albicans ATCC-10231, C. utilis ATCC-

9226 and C. tropicalis ATCC-13803. The test compounds

were soluble in dimethyl sulfoxide (DMSO), in accordance

with the Antimicrobial Susceptibility Testing Protocols.

Hence, the antifungal assays were performed with this

solvent.

Methyl 2-(4-(hydroxymethyl)-2-methoxy-6-nitrophenoxy)

acetate (7c)

Under N₂atmosphere and at 0 °C, a mixture of 7a (1.000 g,

3.71 mmol) and NaBH₄(0.070 g, 1.85 mmol) in MeOH

(10 mL) was stirred for 2 h before removing the solvent

under vacuum. The residue was puriﬁed by column chro-

matography over silica gel (20 g/g of crude, hexane/EtOAc,

1:1) to provide 7c (0.86 g, 86%) as a white solid. Rf 0.16

(hexane/EtOAc, 1:1); m.p. 97–98 °C. IR (KBr): ν 3481,

2921, 1747, 1535, 1452, 1390, 1355, 1276, 1241, 1208,

1135, 1062, 972, 919, 850, 810, 780 cm⁻^{1 1}H NMR

.

(500 MHz, CDCl₃): δ 2.47 (br, 1H, OH), 3.79 (s, 3H,

CO₂CH₃), 3.90 (s, 3H, OCH₃), 4.68 (s, 2H, CH₂OH), 4.73

(s, 2H, CH₂CO₂CH₃), 7.14 (d, J = 1.5 Hz, 1H, H-3′), 7.29

(d, J = 1.5 Hz, 1H, H-5′). ¹³C NMR (125 MHz, CDCl₃): δ

52.2 (CO₂CH₃), 56.4 (OCH₃), 63.7 (CH₂OH), 69.6

(CH₂CO₂CH₃), 113.7 (C-5′), 114.2 (C-3′), 138.1 (C-4′),

139.2 (C-1′), 144.5 (C-6′), 153.3 (C-2′), 169.1 (CO₂CH₃).

HRMS (EI): m/z calcd for C₁₁H₁₃NO₇: 271.0692; found:

271.0687.

The minimum inhibitory concentration (MIC) of all

compounds and the reference drug (itraconazole), expressed

in µg/mL, was determined in 96-well plates, utilizing

RPMI-1640 culture medium buffered with 3-(N-morpho-

lino)propanesulfonic acid (MOPS, Sigma) and dilutions of

16 to 0.03 µg/mL (according to the procedure outlined in

the instructions for each type of fungus).

Hypolipidemic activity

The inoculum for the yeasts was prepared with culture

colonies of 24-h growth strain in SDA medium and resus-

pending them in a tube of saline solution (0.85% NaCl) to

obtain a concentration of ~1–5 × 106 CFU/mL. The neces-

sary amount of saline solution was added to adjust the

optical density to 0.5 McFarland. Subsequently, a 1:1000

dilution was made with RPMI medium. For ﬁlamentous

fungi, the inoculum was elaborated from a culture having

undergone 7 days of growth at 35 °C on PDA agar, a

medium that induces the formation of conidia or spor-

angiospores. To remove the conidia, the culture was

The hypolipidemic effect of the compounds was studied on

male ICR mice weighing 25–30 g (Birmex, Mexico City),

which were housed in hanging metal cages and maintained

at 24 2 °C and 50 10% relative humidity on a 12 h light/

dark cycle (lights on at 8:00 a.m.). Food (a standard pellet

diet, Rodent Diet 5001, PMI Nutrition International, Bren-

wood, MO) and water were freely available. All animals

appeared healthy throughout the dosing period, maintaining

normal food intake and weight gain. Post-sacriﬁce analysis

Medicinal Chemistry Research

introduced and handled in Tween 20, then resuspended in

saline solution. The particles were allowed to settle for

5 min, the supernatant transferred to another tube and sha-

ken vigorously for 15 s. Because the size of the conidia is

different for each species, the optical density required to

each well was measured on an ELISA microplate reader at

595 nm. The result was expressed as the percentage of the

viability of control cells and the LC₅₀was calculated (van

Meerloo et al. 2011).

reach

a

concentration of 1–5 × 10⁶CFU/mL varied.

Data analysis

Adjustment to the desired value was accomplished with the

McFarland Turbidity Standard No. 0.15, based on the

methods of the CLSI.

Data analysis was carried out by nonparametric one-way

ANOVA and a post hoc with Dunn’s test, run on Sigma Plot

software version 12 (Motulsky 1999). All data are expressed

as the mean SD, with signiﬁcance set at p < 0.05. Each

experiment was performed in triplicate.

Compounds 5–7 and 9 were prepared (at 10 mM) and

tested for their inhibition of Fusarium oxysporum with the

PDA dilution method (Lalitha 2004). Brieﬂy, PDA medium

was sterilized in an autoclave at 121 °C for 20 min, fol-

lowed by the addition of one of the compounds diluted in

DMSO (2%). The plates were inoculated with F. oxysporum

and then incubated at 26–28 °C for 72 h. All experiments

were carried out in triplicates (n = 12), and the data are

expressed as the mean. The growth inhibition efﬁciency of

each compound was compared with captan (10 mM), the

reference drug, and validated by ANOVA and the Tukey

test (p < 0.05) on the SAS V9.0 program (SAS 2014).

Antioxidant activity

DPPH radical scavenging assay

The scavenging of free radicals by the chromones was

assessed by using the 2,2-diphenyl-1-picrylhydrazyl radical

(DPPH^•) assay, as previously described (Cevallos-Casals

and Cisneros-Zevallos 2003) with slight modiﬁcations. A

concentration of 10 mM of each compound was prepared, to

which a solution of DPPH^•(133.33 µM) was added at a ratio

of 1:3 (v/v). The mixture was incubated at 37 °C for 30 min

and read at 517 nm. Scavenging capacity (SC) was

expressed as the percentage decrease in DPPH at 10 mM:

Cytotoxic activity

Cell culture

Cell lines of human cervical cancer (HeLa), prostate cancer

(DU-145), breast cancer (MDA-231), and no tumoral cell line

(HaCaT) were cultivated in Dulbecco’s Minimum Essential

Medium (DMEM) containing 10% fetal bovine serum (FBS)

and 1% penicillin/strectomycin (100/100 U/mL). Cells were

cultured as adherent monolayers and maintained at 37 °C

and 5% humidity (Freshney 2010).

SC% ¼ ½ðA_controlÀ A_testÞ=A_control Â 100

where A_controlis the absorbance of the DPPH solution

(control) and A_testis the absorbance of the DPPH solution

plus a compound.

ABTS radical activity

Cell viability assay

The free radical scavenging capacity was quantiﬁed based

on a slightly modiﬁed version of the previously reported

Cell viability was determined by an MTT (3-[4,5-dime-

thylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay.

Cells were harvested, counted, and transferred to 96-well

plates at 3000 cells per well to be incubated for 24 h. Upon

completion of this time, cells were treated with compounds

5a–b, 6a–b, and 9a–c at concentrations of 10, 20, 40, 60,

80, and 100 μg/mL with 1% DMSO in cell culture medium,

then again incubated for 24 h. Subsequently, microscope

images were obtained for each cell line exposed to each

compound. Cells without treatment were used as the via-

bility control, 1% DMSO as the vehicle control, and

methotrexate as the control for comparison of the respective

median lethal concentrations (LC₅₀).

2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic

acid)

radical cation (ABTS^•+) assay (Gallardo et al. 2016; Zhang

et al. 2013). The radical cation was prepared by dissolving

the stock solution of ABTS (7 mM in distilled water) with

K₂S₂O₈solution (2.45 mM) at ratio of 1:1 (v/v) and then

leaving the mixture to stand in the dark at rt for 16 h. For the

evaluation of antioxidant activity, the ABTS^•+solution was

diluted with absolute ethanol until reaching the absorbency

of 0.700 0.02 at 734 nm. Taking 10 µL of the resulting

solution at different concentrations (10, 1.0 and 0.1 mM),

1 mL of ABTS solution was added and after 6 min absor-

bance was read at 734 nm. All tests were performed in tri-

plicate and the mean was centered. Finally, the percentage

inhibition of ABTS absorbance was calculated by the

aforementioned formula for the DPPH^•assay. The data are

expressed as Trolox equivalent antioxidant capacity (TEAC),

developing a standard curve in a range of 0.5 to 3.5 mM.

In each well, 50 μL of a solution of MTT in culture

medium (100 μg/μL) were added and cells were incubated at

37 °C for 4 h. The culture medium was removed and 100 μL

of isopropanol was added to each well. Optical density in

Medicinal Chemistry Research

Molecular docking study

Results and discussion

Protein and ligand structures

Chemistry

The crystallographic structures of human HMGRh in

complex with a selective inhibitor (simvastatin) were

retrieved from the protein data bank (PDB) (http://www.

rcsb.org/) with the code 1HW9 (Istvan and Deisenhofer

2001). Before docking simulations were run, the coordi-

nates of the protein were set, water molecules removed,

hydrogen atoms added to the polar atoms (considering pH at

7.4), and Kollman charges assigned. The 3D structures of

simvastatin were downloaded from the Zinc database (Irwin

and Shoichet 2005). The structures of the ligands were

sketched in 2D with ChemSketch (https://www.acdlabs.

com/resources/freeware/chemsketch/) and optimized with

AM1 on Gaussian 98 software (Frisch et al. 2004) to obtain

the minimum energy conformation for docking studies.

The synthesis of the series of methyl phenoxyacetates 5a–c

and 6a–c was based on the functionalization of vanillin (8)

as the starting material (Scheme 1). Firstly, the hydroxy

group was protected by the acetate moiety through a Wil-

liamson alkylation with methyl bromoacetate under basic

conditions (Argüelles et al. 2010) to give compound 10 in

high yield. The conversion of the formyl group to the

hydroxy group was carried out by a Baeyer–Villiger rear-

rangement with mCPBA, followed by hydrolysis of the

formyl ester (Argüelles et al. 2010), to afford the phenol

derivative 11 in 72% yield. Acylation of the latter in the

ortho position of the hydroxy group was achieved by a

Lewis acid (BF₃·OEt₂)-catalyzed reaction (probably through

a Fries rearrangement) in the presence of the acid chloride

to furnish the corresponding acylphenoxyacetic esters 5a–c

in fairly good yields (Scheme 1). Conversion of this series

into the alkylphenoxyacetic esters 6a–c took place through

a Clemmensen reduction to provide the expected products

in good yields.

Molecular docking

The protein-ligand interaction was observed on Autodock

version 4.0 and AutoDockTools (Morris et al. 2009). All the

possible rotatable bonds, torsion angles, atomic partial

charges and non-polar hydrogens were determined for each

ligand. For HMGRh, the grid dimensions in Auto-

DockTools were 62 × 82 × 106 Å³with points separated by

0.375 Å, centered at X = 5.564, Y = −6.681, and Z = 4.637.

The hybrid Lamarckian Genetic Algorithm was applied for

minimization, utilizing default parameters. A total of one

hundred docking runs were conducted, adopting the con-

formation with the lowest binding energy (kcal/mol) for all

further simulations. The script and ﬁles were prepared and

the docking results visualized on AutoDockTools, then

edited in Discovery 4.0 Client (Dassault Systèmes 2016).

1

The structures of all compounds were established by H

NMR, ¹³C NMR, and HRMS. The full assignment of proton

and carbon signals was achieved by 2D NMR experiments

(HMQC and HMBC). The selective formation of the tet-

rasubstituted benzene ring was ascertained by the two

characteristic singlets for the para aromatic protons. The

presence of the carbonyl group of the series of acyl deri-

vatives 5a–c was veriﬁed by the signal appearing at around

204 ppm in the ¹³C NMR spectra.

The series of nitro compounds 7a–c was readily prepared

by direct nitration of vanillin (8) to provide the known 5-

nitrovanillin (12) (Grenier et al. 2000; Kiss et al. 2010;

Scheme 1 Synthesis of phenol 10, acyl phenols 5a-c, and alkyl phenols 6a-c

Medicinal Chemistry Research

Scheme 3 Synthesis of 1,2-dihydroquinolines 9a-d

Scheme 2 Synthesis of methyl nitrophenoxyacetates 7a-c from

vanillin (8)

(tyloxapol), a non-ionic surfactant (Levine and Saltzman

2007; Zeniya and Reuben 1988; Edelstein et al. 1985). It

causes a drastic rise in serum triglycerides and cholesterol

levels by increasing HMGR activity (Kuroda et al. 1977;

Goldfard 1978). To diminish the concentration of blood

serum lipids, it is necessary to inhibit the synthesis of endo-

genous cholesterol (elevated after treatment with tyloxapol),

limit the absorption of lipoprotein lipids, and stimulate the

excretion of the latter (Korolenko et al. 2010). Many clinically

used drugs accomplish some of these functions, such as

ezetimibe (inhibitor of cholesterol absorption), ﬁbrates

(PPAR-α agonists) and statins (HMGR inhibitors).

The series of analogs 5a–c, 6a–c, 7a, and 9a–d were

administered ip at doses of 25, 50, and 100 mg/kg to

hyperlipidemic mice, and their effect on the plasma levels

(mg/dL) of total cholesterol, and triglycerides was examined

with a procedure similar to that employed in previous studies

with related compounds (Argüelles et al. 2010). Since pre-

liminary assays showed a very modest decrease in cholesterol

by 7a–c, and considering that the nitro group can be cytotoxic

(Chung et al. 2011; Olender et al. 2018), only 7a was sub-

jected to further assessment of hypolipidemic activity.

All compounds evaluated, including the reference (sim-

vastatin), sharply depleted plasma levels of cholesterol and

triglycerides, in the range of 44–75% and 51–93%,

respectively, vs. the hyperlipidemic control animals. A

signiﬁcant difference existed between the values of the

control and most treatments (Table 1). Moreover, it is worth

noticing that almost all compounds signiﬁcantly decrease

triglyceride levels, since it is critical to diminish the con-

centration of the latter because in excess they induce the

formation of atheroma, leading to an imminent cardiovas-

cular disease (Veseli et al. 2017).

Rashid et al. 2018) (Scheme 2). The latter was treated with

methyl bromoacetate under basic conditions to obtain the

nitrophenoxyacetate 7a in a 67% yield for the two steps.

Baeyer–Villiger rearrangement produced the respective

formate, which was hydrolyzed under acid conditions to

deliver 7b in 61% yield. Moreover, reduction of the formyl

group of 7a with sodium borohydride resulted in the

hydroxymethyl derivative 7c in 86% yield.

The synthesis of 1,2-dihydroquinolines 9a–d was

achieved through a single-step reaction, according to the

published procedure (Gutiérrez et al. 2013) (Scheme 3).

Thus, analogs 9a–b were prepared in high yields by react-

ing m-anisidine (13a) with ketones 14a–b, under solvent-

free conditions and in the presence of magnesium bromide

as the catalyst and lithium carbonate as the base. Mean-

while, 1,2-dihydroquinoline 9c was formed by using p-

anisidine (13b) and methyl pyruvate (14c) under similar

reaction conditions. Finally, the spiro-polycyclic 1,2-dihy-

droquinoline 9d was furnished in high yield via the reaction

of 13a with cyclopentanone (14d). The physical data and

NMR spectra of the resulting 1,2-dihydroquinolines were in

agreement with the previous report (Gutiérrez et al. 2013).

Hypolipidemic activity

The in vivo hypolipidemic screening of the compounds 5a–c,

6a–c, 7a, and 9a–d was performed on male ICR mice sub-

jected to tyloxapol-induced hyperlipidemia (Silva et al. 2001;

Kourounakis et al. 2002). The possible mechanism of lipid-

lowering agents is commonly evaluated with Triton WR 1339

Medicinal Chemistry Research

Table 1 Effect of the test compounds on the serum lipid proﬁle of

derived from isovanillin 4a–f (Mendieta et al. 2014), as well

as for the analogs to α-asarone 3a–f (Mendieta et al. 2014).

However, an inverse hypolipidemic effect was found for

other homologs structurally closer to α-asarone (Cruz et al.

2001), among which the most potent activity was displayed

by those containing an alkyl vs. acyl side chain.

Actually, the alkyl side chain would be expected to act as

an important hypocholesterolemic pharmacophore, because

of its strong lypophilic binding interaction with the HMGR

active site (Argüelles et al. 2010). Nevertheless, there was

no relation between the side chain length and the hypo-

cholesterolemic effect, evidenced by the comparable

reduction in serum cholesterol values of the homologs in the

5a–c and 6a–c series. Regardless of the possible synergetic

effect of the acyl and alkyl side chains on the test com-

pounds, the phenoxyacetic ester frame appears to play the

main role as the pharmacophore moiety (Table 1), as has

been reported (Argüelles et al. 2010; Cruz et al. 2003;

Labarrios et al. 1999; Zúñiga et al. 2005; Mendieta et al.

2014).

Interestingly, among the four tested 1,2-dihy-

droquinolines, 9a, 9b, and 9d displayed high hypocholes-

terolemic and hypotriglyceridemic activity, which is

relevant and almost unique for this kind of heterocycles

(Lagu et al. 2007; Matsuda et al. 2007; Guo et al. 2017).

The change of substituents in the A ring of the heterocyclic

frame does not seem to have a signiﬁcant inﬂuence on the

hypolipidemic effect of the compounds, suggesting that the

1,2-dihydroquinoline scaffold could be the active pharma-

cophore of these potential hypolipidemic agents. Despite

the non-signiﬁcant hypocholesterolemic activity of 9c, it

was able to diminish the level of triglycerides signiﬁcantly.

Although a dose-response relationship cannot be estab-

lished for each member of the series, in many cases the

25 mg/kg dose (the closest to that of the control group)

produced the greatest effect (Table 1).

male ICR mice^a

Compound

Dose mg/ Cholesterol

Kg/day

Triglycerides

Normal diet

Simvastatin

Tyloxapol

−78.15 0.07* −80.02 0.08*

−76.38 0.19* −79.10 0.17*

17

400

100 1.03^b

100 0.33^c

5a + Tyloxapol 25

−67.29 6.78* −83.74 2.44*

−75.44 6.27* −93.40 1.25*

−58.5 7.35* −86.46 2.31*

−75.20 3.52* −81.42 2.51*

−57.49 7.74* −87.49 1.25*

−52.53 11.61* −88.95 1.31*

−73.43 4.70* −92.04 2.51*

−64.93 11.06* −91.4 1.79*

−74.02 5.14* −85.70 1.84*

−72.84 0.25* −20.77 0.79

−74.73 0.09* −80.59 0.11*

−65.40 0.69* −73.95 0.38*

50

100

5b + Tyloxapol 25

50

100

5c + Tyloxapol 25

50

100

6a + Tyloxapol 25

50

100

6b + Tyloxapol 25

−69.89 0.49*

−61.03 0.60*

−70.89 0.58*

4.11 1.14

4.11 2.18

50

100

−9.55 0.83

6c + Tyloxapol 25

−71.07 0.70* −76.25 0.34

−69.30 0.30* −75.72 0.19*

−72.72 0.41* −78.22 0.29*

50

100

7a + tyloxapol 25

−58.43 0.53

−59.03 0.78

−17.46 2.68

−67.13 0.40*

−66.98 0.60*

−43.01 0.37

50

100

9a + tyloxapol 25

−72.27 0.11* −76.22 0.24*

−63.16 0.62* −63.26 0.62*

−72.72 0.24* −70.47 0.44*

−68.82 0.29* −67.37 0.55*

−44.98 1.45* −34.11 0.54

−73.78 0.33* −75.25 0.22*

50

100

9b + tyloxapol 25

50

100

9c + tyloxapol 25

−51.70 1.01

28.22 7.82

−65.99 0.4

−60.13 0.46*

−66.94 0.57*

−82.79 0.20*

50

Antifungal activity

100

9d + tyloxapol 25

−72.72 0.42* −73.92 0.21*

−63.99 0.72* −61.10 0.75*

−62.01 0.39* −51.36 0.39*

Statins have been reported to produce in vitro activity

against several human pathogenic fungi, including Candida

spp. and Aspergillus spp. (Qiao et al. 2007; Cabral et al.

2013). Hence, the test compounds (except 6c and 7b) and

itraconazole (the reference) were tested in vitro against four

ﬁlamentous fungi (Aspergillus fumigatus ATCC-16907,

Trichosporon cutaneum ATCC-28592, Rhizopus oryzae

ATCC-10329, and Mucor hiemalis ATCC-8690) and three

yeast specimens (Candida albicans ATCC-10231, C. utilis

ATCC-9226, and C. tropicalis ATCC-13803) (Table 2). The

MIC values (expressed in μg/mL) of all compounds were

determined in 96-well plates with 3-(N-morpholino)propa-

nesulfonic acid (MOPS) as the buffer. Standardized micro-

biological methods developed by the CLSI (Fothergill 2012;

50

100

*Signiﬁcantly different from the tyloxapol group (p < 0.05)

^aExpressed as a percentage of the group treated with tyloxapol only

(mean standar error, n = 6)

^b141.16 mmol/L

^c1.26 mmol/L

The best hypolipidemic agents were the acylphenox-

yacetic esters 5a–c, depleting cholesterol and triglycerides

to almost the same extent as the positive control (simvas-

tatin). A similar result was observed for the analogous series

Medicinal Chemistry Research

Table 2 In vitro antifungal

activities of most synthesized

compounds (MIC, μg/mL)

Compound

Yeast fungi

Filamentous fungi

C. alb.

C. uti.

C. trop.

M. hie.

A. fum.

R. ory.

T. cut.

5a

8

4

16

8

16

1

16

8

5b

8

16

2

5c

2

16

8

16

4

8

6a

0.5

8

16

1

6b

16

8

7a

16

8

7c

4

16

8

16

8

9a

4

16

8

9b

16

4

16

4

1

9c

8

16

1

9d

Standard^a

1

16

0.06

0.03

0.25

C. alb. Candida albicans, C. uti. Candida utilis, C. trop. Candida tropicalis, M. hie Mucor hiemalis, A. fum

Aspergillus fumigatus, R. ory Rhizopus oryzae, T. cut Trichosporon cutaneum

^aItraconazole

La Regina et al. 2008; Ramírez-Villalva et al. 2017; García-

Vanegas et al. 2019) were employed. The sensitivity of the

ﬁlamentous microorganisms was determined by the micro-

dilution M38-A method (CLSI 2002; Espinel-Ingroff and

Canton 2007a), and that of the yeast fungi with the M27-A3

method (CLSI 2008; Espinel-Ingroff and Cantón 2007b;

Pfaller and Diekema 2012).

A good effect was produced against C. albicans by 6a

and 6b, though not as good as the inhibition elicited by

itraconazole. Contrarily, a lower antifungal activity was

found for 5c and 9d. None of the present derivatives were

active against the yeast fungi C. utilis and C. tropicalis.

Compared with the effect of itraconazole, only 6b and 9b

demonstrated good growth inhibition of the ﬁlamentous

fungus R. oryzae, and 5a and 7a of T. cutaneum.

No structure-activity correlation was detected among the

active compounds in regard to the growth inhibition of C.

albicans. However, homologs 6 (containing C-5 alkyl

groups) gave a greater antifungal effect than homologs 5

(with the C-5 acyl moiety). Overall, these results indicate

that the presence of the methyl phenoxyacetate and 1,2-

dihydroquinoline scaffolds seems to impact antifungal

capacity.

Fusariosis is one of the most common infections in

humans (Garnica and Nucci 2013; Guarro 2013). Fusarium

oxysporum is an invasive pathogen known to cause infec-

tions like keratitis and onychomycosis in immunocompetent

individuals, but in immunocompromised patients the

infections are frequently fatal (Garnica and Nucci 2013;

Guarro and Gené 1995; Cordoba-Guijarro et al. 2002;

Olivares et al. 2005). Although the optimal treatment

against fusariosis is unclear, the modestly effective drugs

consist of antifungal azoles and amphotericin B. In vitro

data demonstrates general resistance of Fusarium to the

available antifungal drugs with poor MIC values (Guarro

2013; Cordoba-Guijarro et al. 2002).

Consequently, the series 5–7 and 9 were tested as inhi-

bitors of the mycelial growth of the pathogen F. oxysporum,

using the agar dilution method and captan as the positive

control. Captan, a fungicide utilized in agriculture, cos-

metics and pharmaceuticals (U.S. Environmental Protection

Agency 1984) inhibits the mycelial growth of Fusarium

spp. (Türkkan and Erper 2015). According to previous

studies, it also increases cell vulnerability to oxidative stress

(Moreno-Aliaga et al. 1999; Inoue et al. 2018).

The compounds evaluated induced signiﬁcant cell

growth inhibition, from 72 to 90% for 6a–c, and 70% for 9a

and 9c (Fig. 3). Interestingly, the presence of the alkyl side

chain of derivatives 6 signiﬁcantly improved growth inhi-

bition, with 6b displaying the best result (90%). 1,2-Dihy-

droquinolines 9a and 9c contain either an alkyl side chain or

alkyl carboxylate.

Cytotoxic activity

The cytotoxic effect of 5a–b, 6a–b, and 9a–c was deter-

mined through the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-

diphenyltetrazolium bromide) cell viability assay with cell

lines of human cervical cancer (HeLa), prostate cancer (DU-

145), breast cancer (MDA-231) and normal skin keratino-

cytes (HaCaT) (van Meerloo et al. 2011). Death was

observed in up to 90% of the cells exposed to these com-

pounds in concentrations higher than 40 μg/mL. In the

MDA-231 and DU-145 cell lines, there was up to 50% cell

death at 80–100 μg/mL. HeLa and HaCaT cells showed

100% viability following exposure to the non-active

Medicinal Chemistry Research

100.0

90.0

80.0

70.0

60.0

50.0

40.0

30.0

20.0

10.0

0.0

Fig. 3 Growth inhibition of F.

oxysporum produced by 5a–c,

6a–c, 7a, 7c, and 9a–d (at

10 mM) after 72 h of incubation.

Data are expressed as the mean

standard deviation (n = 12)

and are signiﬁcantly different

from the control (p < 0.05)

90.0

86.9

72.1

69.9

70.2

61.6

46.4

43.3

30.3

25.5

23.5

22.8

11.1

captan

5a

5b

5c

6a

6b

6c

7a

7c

9a

9b

9c

9d

Table 3 Median lethal

Compound

Cell line/values

MDA-231

DU-145

HeLa

HaCaT^a

concentration (LC₅₀) (μg/mL)

calculated for derivatives 6a–b

in the four cell lines evaluated

6a

LC₅₀(IC 95%)

24 (8–40)

27 (19–35)

44 (25–62)

73 (60–85)

>100

19 (11–27)

69 (53–85)

>100

6b

–

Methotrexate^b

>100

A 95% conﬁdential interval (CI 95%) was calculated for each LC₅₀

^aNo tumoral cell line

^bThe reference drug

compounds. In cells incubated exclusively with the vehicle

(DMSO), no death occurred, and the decrease in cell growth

relative to the control was non-signiﬁcant.

of α-asarone (1), which has antioxidant properties (Pages

et al. 2010). Secondly, the 1,2-dihydroquinoline ethoxyquin

is a known antioxidant feed additive (Dorey et al. 2000;

Błaszczyk and Skolimowski 2007; Błaszczyk et al. 2013; de

Koning 2002; Błaszczyk et al. 2006; Błaszczyk and Sko-

limowski 2005; Błaszczyk and Skolimowski 2006).

The antioxidant activity of the series of analogs 5a–c,

6a–c, 7a, 7c, and 9a–d was assessed by using the radical

scavenging method with the 2,2-diphenyl-1-picrylhydrazyl

radical (DPPH^•) and 2,2′-azino-bis(3-ethylbenzothiazoline-

6-sulphonic acid) radical cation (ABTS^•+). Antioxidant

activity was expressed as the percentage of decrease in

DPPH and ABTS (Table 4). Butylated hydroxytoluene

(BHT) was the positive control for the DPPH assay, and the

TEAC (mM) was the unit for expressing the data of the

ABTS assay (Huang et al. 2005).

Only compounds 6a, b exhibited cytotoxicity (Table 3).

Derivative 6a produced an exponential decline in cell via-

bility in all cell lines, including non-tumor cells (HaCaT).

Derivative 6b (at 100 μg/mL) caused greater cytotoxicity in

the cervical carcinoma cell line (HeLa) and HaCaT (control)

than the other cell lines. Hence the activity of these com-

pounds was non-selective, a characteristic that may be

improve by future modiﬁcations in their structure. Their

selectivity should be thoroughly studied in various cell lines

(Xiao et al. 2018) to seek speciﬁcity for tumor cells.

Antioxidant activity

Antioxidants are agents capable of protecting a biological

target against oxidative stress, DNA mutations and cell

damage, helping to prevent many human diseases. For

example, oxidative stress plays a pivotal role in the devel-

opment of diabetes, cancer, and cardiovascular disease

(Pisoschi and Pop 2015). The antioxidant potential was

herein evaluated for the synthesized compounds for two

main reasons that suggest a free radical scavenging capacity

of these derivatives. Firstly, alkylphenols 6a–c are analogs

The 6a–c and 9a–d series showed good radical scaven-

ging capacity in the DPPH assay. The IC₅₀values for 6a–c

were in a narrow range (0.24–0.25 mM) and for 9a–d in a

much wider range (0.12–1.52 mM). Moreover, a stronger

antioxidant effect was detected for 6a–c, 9a, and 9c than

BHT (based on the IC₅₀determination). For 6a–c, the

presence of an alkyl side chain apparently promoted anti-

oxidant activity (Gallardo et al. 2016), while the existence

of an acyl group on the side chain notably diminished it.

Medicinal Chemistry Research

Table 4 DPPH^•and ABTS^•+

cation radical scavenging

activity of 5a–c, 6a–c, 7a, 7c,

and 9a–d^a

Compound

DPPH Scavenging activity

ABTS Scavenging activity

% (10 mM)

(IC₅₀mM)

% (10 mM)

(mM Trolox/g)

5a

5b

5c

40.6 0.60

15.9 0.61

19.1 0.60

95.3 1.82

94.2 1.50

94.2 1.10

0.2 0.32

13.40 0.20

11.68 1.38

13.23 0.71

98.80 0.01

98.61 0.01

98.62 2.39

0.91 0.37

0.25 0.48

97.81 3.94

97.53 0.01

98.78 0.01

68.07 0.044

–

0.244 0.002

0.220 0.014

0.216 0.007

1.509 0.001

1.424 0.001

1.349 0.001

0.050 0.004

0.058 0.005

1.552 0.039

1.095 0.001

1.244 0.001

1.00 0.021

–

6a

6b

6c

0.241 0.009

0.255 0.016

0.244 0.012

7a

7c

0.9 0.24

9a

9b

9c

93.5 1.94

93.6 2.49

94.3 1.16

91.0 1.93

85.02 3.33

0.127 0.030

1.519 0.023

0.238 0.022

1.52 0.015

0.84 0.08

9d

BHT

^aThe DPPH scavenging activity is calculated as the IC₅₀, and the ABTS scavenging activity as the mM

Trolox equivalent (TEAC, mM). Data are expressed as the mean standard deviation (n = 6), with

signiﬁcant differences considered at p < 0.05

This characteristic was not affected by the side chain length.

Regarding the 1,2-dihydroquinolines 9a–d, the presence of

a hindered group, such as the phenyl (9b) or the cycloalkyl

(9d) groups, played a negative role, slightly reducing the

DPPH^•scavenging activity (IC₅₀for both 9b and 9d was

1.52 mM). Contrarily, the alkyl (9a) and methoxycarbonyl

(9c) groups improved the antioxidant effect, being almost

sevenfold greater with 9a (IC₅₀= 0.12 mM) than BHT

(IC₅₀= 0.84 mM). The signiﬁcant activity of 9a can be

attributed to the lipophilicity of the 2-ethyl and 2-methyl

groups (Dorey et al. 2000). Interestingly, the methoxy

group attached at either the C-6 or C-7 position of the

quinoline ring did not substantially affect the DPPH^•radical

scavenging capacity, although its presence seems to have

contributed considerably to the antioxidant effect (Dorey

et al. 2000; Błaszczyk and Skolimowski 2007; Błaszczyk

et al. 2013; de Koning 2002).

2014). Hence, a docking study was carried out for the most

active hypolipidemic derivatives (5b, 6a, 9a, and 9d) of the

four series herein evaluated.

Docking studies were conducted on Autodock version

4.0 and AutoDockTools (Morris et al. 2009) to explore the

binding mode of the test compounds and the reference drug

(simvastatin) at the active site of HMGRh (retrieved from

the PDB; code: 1HW9) (Istvan and Deisenhofer 2001). The

binding energy was calculated and the interaction residues

identiﬁed in each case (Table 5).

The binding modes of 5b, 6a, 9a, 9d, and simvastatin

(Fig. 4) involve the amino acid side chains of the active site

of the enzyme, including Glu559, Arg590, Asp690,

Lys691, and Asn755. In a previous study by our group

(Mendieta et al. 2014), these residues were identiﬁed and

Glu559 and Lys691 were found to be the key residues

(Andrade-Pavón et al. 2017; Andrade-Pavón et al. 2019).

The acylphenoxyacetic ester 5b had a better binding energy

(−7.47 kcal/mol) than the alkylphenoxyacetic ester 6a

(−5.93 kcal/mol). The docking data correlate with the

results of the in vivo assessment of their hypolipidemic

activity, since 5a–c decreased the levels of cholesterol and

triglycerides to a greater extent than 6a–c.

Compounds 5b, 6a, and simvastatin share key hydro-

philic interactions, such as conventional hydrogen and

carbon-hydrogen bonding to the active site of the HMGRh

enzyme, mainly with amino acid side chains Glu559,

Ser684, Asp690, and Lys735. There are some key simila-

rities in the interactions. For instance, the hydroxyl group at

C-4 of the aromatic ring of both 5b and 6a interact with the

same residue (Ser684) as the carboxylate group of sim-

vastatin. Moreover, there is an interaction with Lys735 by

Series 6a–c and 9a–d also displayed good ABTS^•+

radical inhibition (Table 4) with 6a and 9a giving the best

ABTS^•+scavenging capacity (1.51 and 1.55 mM of TEAC).

There was a similar pattern for the DPPH^•radical scaven-

ging activity, ﬁnding the best results for 9a and no effect for

compounds 5a–c, 7a and 7c. In summary, the phenoxyacetic

ester and 1,2-dihydroquinoline scaffolds demonstrated a

signiﬁcant antioxidant activity, which was improved by the

presence of an alkyl side chain or a polar carboxylate group.

Docking of analogs 5, 6, and 9 on human HMGR

The in vitro inhibition of HMGRh has been demonstrated

for 3 and 4, showing a signiﬁcant structure-binding contact

similarities with respect to simvastatin (Mendieta et al.

Medicinal Chemistry Research

Fig. 4 Predicted binding mode of simvastatin (a), 5b (b), 6a (c), 9a

(d), and 9d (e) at the active site of HMGRh (1HW9) using Autodock

version 4.0 and AutoDockTools. The 3D model portrays select amino

acid residues bound by the ligands at the active site of HMGRh. Only

the hydrophilic bonds are shown for better clarity. In the 2D model, the

following interactions are denoted with dotted lines: conventional

hydrogen bonds (green), carbon-hydrogen (yellow), alkyl (light pink),

π-anion (brown), π-cation (orange), π-sigma (purple), and π-alkyl

(dark pink). The amino acid residues are illustrated as: hydrophobic

(green), polar (cyan), positively charged (blue), and negatively

charged (red)

Medicinal Chemistry Research

Fig. 5 A1 Overlay of the

docking poses of 5b (red) and 6a

(yellow). B1 Overlay of 9a

(purple) and 9d (green). A2–B2

The binding mode is compared

between the two pairs of

compounds and

simvastatin (blue)

the hydroxyl group at the C-4 position of 6a, the carbonyl

groups of 5b and simvastatin. The three compounds also

have comparable hydrophobic interactions, such as π-sigma

with the amino acid Leu853. Due to their strongly activated

aromatic ring, compounds 5b and 6a share electrostatic

interactions of the π-cation type with the Arg590 residue.

The carbonyl oxygen atom of 5b forms an additional

hydrogen bond contact with the side chain of Lys735. This

residue may affect the enzymatic activity of 6a by inter-

acting with the alkyl side chain. The latter observations are

in agreement with the in vivo assessment of hypolipidemic

activity, which was improved by the polar-induced effect of

the carbonyl group in the chain.

The results support our hypothesis that by replacing the

C-4 methoxy group of 1 by a hydroxyl group, and main-

taining a hydrogen bond network comparable to the one

exiting in 3 and 4, the hypolipidemic effect of 5b and 6a

should be conserved.

For 1,2-dihydroquinolines 9a and 9d, on the other hand,

a very similar binding energy was found (−8.22 and

8.21 kcal/mol, respectively) (Table 5). Furthemore, 9a had

the binding energy closest to the value for simvastatin. It is

likely that the hydrocarbon portion present in both 9a, 9d

and the reference drug enhances the number of the inter-

actions within the active site as well as the overall stability

(Figs 4 and 5). In addition, 9a and 9d share hydrophilic

interactions between their polar NH and the Asp690 residue

of the enzyme, involving conventional hydrogen bonds and

carbon-hydrogen bonds. These interactions are comparable

to those observed for the acylphenoxyacetic esters 5, the

alkylphenoxyacetic esters 6, and the hydroxy group of

simvastatin. The presence of a high electron density aro-

matic ring in 9a and 9d led to π-cation interactions with

Arg590, and to π-alkyl and alkyl interactions with the side

chains of Met657 and Leu857, respectively.

Compounds 5b and 6a adopt similar orientation at the

active site of the enzyme, especially for the activated

aromatic ring (Fig. 5). The orientation of 1,2-dihy-

droquinolines 9a and 9d is also alike, mainly due to the

benzoheterocyclic frame. They adopt a conformation in

which the polar and the hydrophobic functional groups

occupy a position similar to some of the groups in sim-

vatatin with comparable polarity, although the structure

of the latter is different. Despite this difference, com-

pounds 5b, 6a, 9a, and 9d interact with most amino acids

in the active site of the enzyme that are targeted by the

reference drug. Based on the docking data, 5a–c, 6a–c,

and 9a–d are likely to have the same mechanism of action

as simvastatin.

To the best of our knowledge, there are no reports of

molecular docking of 1,2-dihydroquinolines like 9a and 9d

in the active site of the HMGRh enzyme. The interaction

energy calculated from docking was correlated with the

in vivo hypolipidemic effect.

Medicinal Chemistry Research

Conclusions

Compliance with ethical standards

Conﬂict of interest The authors declare that they have no conﬂict of

interest.

Most members of the three series of acyl phenols (5a–c) and

alkyl phenols (6a–c) and 1,2-dihydroquinolines (9a–d)

showed a potent hypolipidemic effect, resulting in an over

67% decrease in serum cholesterol and triglycerides at the

lowest doses (25 g/kg). Using the most effective derivatives

(5b, 6a, 9a, and 9d), docking studies were carried out on

human HMGR ﬁnding that the different polar and non-polar

functional groups of the ligands exhibited strong and mul-

tiple interactions with the active site of this enzyme. Hence,

the mechanism of action of these compounds probably

involves the inhibition of HMGR. However, due to the

structural similarity of analogs 5 and 6 with ﬁbrates, the

activation of PPARα cannot be ruled out as a competitive

mechanism participating in a reduction of the level of

triglycerides.

Publisher’s note Springer Nature remains neutral with regard to

jurisdictional claims in published maps and institutional afﬁliations.

References

Aguilar-Salinas CA, Olaiz G, Valles V, Ríos-Torres JM, Gómez-Pérez

FJ, Rull JA, Franco A, Sepulveda J (2001) High prevalence of

low HDL cholesterol concentrations and mixed hyperlipidemia in

a Mexican nationwide survey. J Lipid Res 42:1298–1307

Andrade-Pavón D, Cuevas-Hernández RI, Trujillo-Ferrara JG, Her-

nández-Rodríguez C, Ibarra JA, Villa-Tanaca L (2017) Recom-

binant 3-hydroxy 3-methyl glutaryl-CoA reductase from Candida

glabrata (Rec-CgHMGR) obtained by heterologous expression, as

a novel therapeutic target model for testing synthetic drugs. Appl

Biochem Biotechnol 182:1478–1490

Regarding antifungal potential, the same three series

caused a moderate to robust growth inhibition of C. albi-

cans, R. oryzae, T. cutaneum and F. oxysporum, which are

highly pathogenic fungi. For the latter fungus, derivatives

6a–c and 9a and 9c proved to be more active than the

positive control (captan). On the other hand, derivative 6a

produced cytotoxicity not only in all the cancer cell lines

(MDA-231, DU-145 and HeLa) but also in normal cells

(HaCaT), indicating a lack of selectivity. Finally, both the

6a–c and 9a–d series demonstrated good antioxidant

capacity, evidenced by the strong DPPH and ABTS

scavenging activity. Indeed, 6a–c and 9a and 9c were more

active than the positive control (BHT).

Overall, the phenoxyacetic acid esters derived from

vanillin and 1,2-dihydroquinolines showed promise as

frames for novel hypolipidemic, antifungal, anticancer

and antioxidant agents. Thus, the corresponding struc-

tures may be advantageous as templates for the design of

new compounds with more potent pharmacological

activity.

Andrade-Pavón D, Ortiz-Álvarez J, Sánchez-Sandoval E, Tamariz J,

Hernández-Rodríguez C, Ibarra JA, Villa-Tanaca L (2019) Inhi-

bition of recombinant enzyme 3-hydroxy-3-methylglutaryl-CoA

reductase from Candida glabrata by α-asarone-based synthetic

compounds as antifungal agents. J Biotechnol 292:64–67

Argüelles N, Sánchez-Sandoval E, Mendieta A, Villa-Tanaca L,

Garduño-Siciliano L, Jiménez F, Cruz MC, Medina-Franco JL,

Chamorro-Cevallos G, Tamariz J (2010) Design, synthesis, and

docking of highly hypolipidemic agents: Schizosaccharomyces

pombe as a new model for evaluating α-asarone-based HMG CoA

inhibitors. Bioorg Med Chem 18:4238–4248

BIP study group (2000) Secondary prevention by raising HDL cho-

lesterol and reducing triglycerides in patients with coronary artery

disease. The bezaﬁbrate infarction prevention (BIP) study. Cir-

culation 102:21–27

Błaszczyk A, Augustyniak A, Skolimowski J (2013) Ethoxyquin: an

antioxidant used in animal feed. Int J Food Sci 2013:585931

Błaszczyk A, Skolimowski J (2005) Apoptosis and cytotoxicity caused

by ethoxyquin and two of its salts. Cell Mol Biol Lett 10:15–21

Błaszczyk A, Skolimowski J (2006) Comparative analysis of cyto-

toxic, genotoxic and antioxidant effects of 2,2,4,7-tetramethyl-

1,2,3,4-tetrahydroquinoline and ethoxyquin on human lympho-

cytes. Chem-Biol Interact 162:70–80

Błaszczyk A, Skolimowski J (2007) Preparation of ethoxyquin salts

and their genotoxic and antioxidant effects on human lympho-

cytes. Arkivoc vi:217–229

Acknowledgements We thank Dr Alberto Feliciano and Rsuini U.

Gutiérrez for experimental support and Bruce Allan Larsen for

proofreading the manuscript. AM, LG-S, MCC-L, FJ, OG-G, LV-T,

ER-G, GC-C, FD, and JT gratefully acknowledge the ﬁnancial support

provided by SIP-IPN (grants 20160791, 20170902, 20180198,

20181332, 20181433, 20195228, 20195287, and 20195606) and

CONACYT (grants 178319, 282033, CB-283225, A1-S-17131, and

300520). AF-B and CG-R are thankful to the UAEM for funding.

MAV gratefully recognized the spectroscopy services provided by the

National Laboratory of the Universidad de Guanajuato (UGUAA-

CONACYT grant 260373). AP, AM, DAM, RIH-B, LR, CR-E, AR-

V, JL, BR-A, and AS-J appreciate the graduate scholarship awarded

by CONACYT, as well as the scholarship complements furnished by

the SIP-IPN (BEIFI) and the Ludwig K. Hellweg Foundation. LG-S,

MCC-L, FJ; LV-T, ER-G, GC-C, FD and JT are fellows of the EDI-

IPN and COFAA-IPN programs.

Błaszczyk A, Skolimowski J, Materac A (2006) Genotoxic and anti-

oxidant activities of ethoxyquin salts evaluated by the comet

assay. Chem-Biol Interact 162:268–273

Bradfute DL, Simoni RD (1994) Non-sterol compounds that regulate

cholesterogenesis. Analogues of farnesyl pyrophosphate reduce

3-hydroxy-3-methylglutaryl-coenzyme A reductase levels. J Biol

Chem 269:6645–6650

Brown JW, Gangloff AR, Jennings AJ, Vu PH (2010) Poly (ADP-

ribose) polymerase (PARP) inhibitors. Patent WO 2010/111626 A2

Cabral ME, Figueroa LIC, Fariña JI (2013) Synergistic antifungal

activity of statin-azole associations as witnessed by Sacchar-

omyces cerevisiae- and Candida utilis-bioassays and ergosterol

quantiﬁcation. Rev Iberoam Micol 30:31–38

Campos-Ríos MG, Chiang Cabrera F (2006) Una revisión nomen-

clatural de los tipos de plantas de la península de Yucatán

(México). Polibotánica 89–149

Medicinal Chemistry Research

Cevallos-Casals BA, Cisneros-Zevallos L (2003) Stoichiometric and

kinetic studies of phenolic antioxidants from Anden purple corn

and red-ﬂeshed sweetpotato. J Agric Food Chem 51:3313–3319

Chalasani N (2005) Statins and hepatotoxicity: focus on patients with

fatty liver. Hepatology 41:690–695

Chamorro G, Salazar M, Salazar S, Mendoza T (1993) Farmacología y

toxicologia de Guatteria gaumeri y alfa-asarona. Rev Invest Clin

45:597–604

Field FJ, Born E, Mathur SN (1997) Effect of micellar α-sitosterol on

cholesterol metabolism in CaCo-2 cells. J Lipid Res 38:348

Fothergill AW (2012) Antifungal susceptibility testing: Clinical

Laboratory and Standards Institute (CLSI) methods. In: Hall GS

(ed) Interactions of yeasts, moulds, and antifungal agents. How to

detect resistance. Springer Science-Business Media, Berlin,

p 65–74

Freshney RI (2010) Deﬁne media and supplements. In: Culture of

animal cells, chap. 8. Wiley-Blackwell, Hoboken, NJ, p 99–114

Frisch MJ, Trucks GW, Schlegel HB, Scuseria GE, Robb MA,

Cheeseman JR, Montgomery Jr JA, Vreven T, Kudin KN, Burant

JC, Millam JM, Iyengar SS, Tomasi J, Barone V, Mennucci B,

Cossi M, Scalmani G, Rega N, Petersson GA, Nakatsuji H, Hada

M, Ehara M, Toyota K, Fukuda R, Hasegawa J, Ishida M,

Nakajima T, Honda Y, Kitao O, Nakai H, Klene M, Li X, Knox

JE, Hratchian HP, Cross JB, Bakken V, Adamo C, Jaramillo J,

Gomperts R, Stratmann RE, Yazyev O, Austin AJ, Cammi R,

Pomelli C, Ochterski JW, Ayala PY, Morokuma K, Voth GA,

Salvador P, Dannenberg JJ, Zakrzewski VG, Dapprich S, Daniels

AD, Strain MC, Farkas O, Malick DK, Rabuck AD, Ragha-

vachari K, Foresman JB, Ortiz JV, Cui Q, Baboul AG, Clifford S,

Cioslowski J, Stefanov BB, Liu G, Liashenko A, Piskorz P,

Komaromi I, Martin RL, Fox DJ, Keith T, Al-Laham MA, Peng

CY, Nanayakkara A, Challacombe M, Gill PMW, Johnson B,

Chen W, Wong MW, Gonzalez C, Pople JA (2004) Gaussian 98

Inc., Version A.6, Wallingford, CT

Gallardo E, Palma-Valdés R, Sarriá B, Gallardo I, de la Cruz JP, Bravo

L, Mateos R, Espartero JL (2016) Synthesis and antioxidant

activity of alkyl nitro derivatives of hydroxytyrosol. Molecules

21:E656

García-Vanegas JJ, Ramírez-Villalva A, Fuentes-Benites A, Martínez-

Otero D, González-Rivas N, Cuevas-Yañez E (2019) Synthesis

and in-vitro biological evaluation of 1,1-diaryl-2-(1,2,3)triazol-1-

yl-ethanol derivatives as antifungal compounds ﬂutriafol analo-

gues. J Chem Sci 131:27

Chander S, Ashok P, Zheng Y-T, Wang P, Raja KS, Taneja A,

Murugesan S (2016a) Design, synthesis, and in-vitro evaluation

of novel tetrahydroquinoline carbamates as HIV-1 RT inhibitor

and their antifungal activity. Bioorg Chem 64:66–73

Chander S, Wang P, Ashok P, Yang L-M, Zheng Y-T, Murugesen S

(2016b) Rational design, synthesis, anti-HIV-1 RT and anti-

microbial activity of novel 3-(6-methoxy-3,4-dihydroquinolin-1

(2H)-yl)-1-(piperazin-1-yl)propan-1-one derivatives. Bioorg

Chem 67:75–83

Chung MC, Bosquesi PL, dos Santos JL (2011) A prodrug approach to

improve the physico-chemical properties and decrease the geno-

toxicity of nitro compounds. Curr Pharm Des 17:3515–3526

Clinical and Laboratory Standards Institute (CLSI) (2002) Document

M38-A2: reference method for broth dilution antifungal sus-

ceptibility testing of ﬁlamentous fungi, 2nd edn. Approved

Standard, Clinical and Laboratory Standards Institute, Wayne, PA

Clinical and Laboratory Standards Institute (CLSI) (2008) M27-A3:

reference Method for broth dilution antifungal susceptibility

testing of yeasts, 3rd edn. Approved Standard, Clinical and

Laboratory Standards Institute, Wayne, PA

Cordoba-Guijarro S, Ruiz-Rodríguez R, Acevedo-Barberá A, Serrano-

Pardo R (2002) Disseminated fusarium infection in a patient with

acute myelogenous leukemia. Actas Dermo-Siﬁliogr 93:118–121

Cruz MC, Salazar M, Garcíaﬁgueroa Y, Hernández D, Díaz F, Cha-

morro G, Tamariz J (2003) Hypolipidemic activity of new phe-

noxyacetic derivatives related to α-asarone with minimal

pharmacophore features. Drug Dev Res 60:186–195

Cruz A, Garduño L, Salazar M, Martinez E, Jiménez-Vázquez HA,

Diaz F, Chamorro G, Tamariz J (2001) Synthesis and hypolipi-

demic activity of modiﬁed side chain α-asarone homologues.

Arznei-Forsch/Drug Res 51:535–544

Garnica M, Nucci M (2013) Epidemiology of fusariosis. Curr Fungal

Infect Rep. 7:301–305

Gielen S, Sandri M, Schuler G, Teupser D (2009) Risk factor man-

agement: antiatherogenic therapies. Eur

J Cardiovasc Prev

Dassault Systèmes (2016) BIOVIA, discovery studio modeling

environment; release 2017. Dassault Systèmes, San Diego, CA

Davidson MH, Armani A, McKenney JM, Jacobson TA (2007) Safety

considerations with ﬁbrate therapy. Am J Cardiol 99:S3–S18

de Koning AJ (2002) The antioxidant ethoxyquin and its analogues: a

review. Int J Food Prop 5:451–461

Dorey G, Lockhart B, Lestage P, Casara P (2000) New quinolinic

derivatives as centrally active antioxidants. Bioorg Med Chem

Lett 10:935–939

Edelstein C, Byrne RE, Yamamoto K, Zarins C, Scanu AM (1985)

Plasma lipoprotein changes attending the intravenous adminis-

tration of Triton WR-1339 in normolipidemic dogs: preferential

effect on high density lipoproteins. J Lipid Res 26:351–359

Elisaf MS, Florentin M, Liberopoulos EN, Mikhailidis DP (2008)

Fibrate-associated adverse effects beyond muscle and liver toxi-

city. Curr Pharm Des 14:574–587

Espinel-Ingroff A, Canton E (2007a) Antifungal susceptibility testing

of ﬁlamentous fungi. In: Schwalbe R, Steele-Moore L, Goodwin

AC (eds) Antimicrobial susceptibility testing protocols. CRC

Press, Florida, p 209–241

Rehabil 16:S29–S36

Goldfard S (1978) Rapid increase in hepatic HMG CoA reductase

activity and in vivo cholesterol synthesis after Triton WR 1339

injection. J Lipid Res 19:489–494

Golomb BA, Evans MA (2008) Statin adverse effects: a review of the

literature and evidence for a mitochondrial mechanism. Am J

Cardiovasc Drugs 8:373–418

Graham DJ, Staffa JA, Shatin D, Andrade SE, Schech SD, La Grenade

L, Gurwitz JH, Chan KA, Goodman MJ, Platt RP (2004) Inci-

dence of hospitalized rhabdomyolysis in patients treated with

lipid-lowering drugs. JAMA 292:2585–2590

Grenier JL, Cotelle N, Catteau JP, Cotelle P (2000) Synthesis and

physico-chemical properties of nitrocaffeic acids. J Phys Org

Chem 13:511–517

Grundy SM, Cleeman JI, Daniels SR, Donato KA, Eckel RH, Franklin

BA, Gordon DJ, Krauss RM, Savage PJ, Smith Jr SC, Spertus JA,

Costa F (2005) Diagnosis and management of the metabolic

syndrome. An American Heart Association/National Heart, Lung,

and Blood Institute Scientiﬁc Statement. Circulation

112:2735–2752

Espinel-Ingroff A, Cantón E (2007b) Antifungal susceptibility testing

of yeasts. In: Schwalbe R, Steele-Moore L, Goodwin AC (eds)

Antimicrobial susceptibility testing protocols. CRC Press, Flor-

ida, p 173–208

Fazio S, Linton MF (2004) The role of ﬁbrates in managing hyperli-

pidemia: mechanisms of action and clinical efﬁcacy. Curr

Atheroscler Rep. 6:148–157

Guarro J (2013) Fusariosis, a complex infection caused by a high

diversity of fungal species refractory to treatment. Eur J Clin

Microbiol Infect Dis 32:1491–1500

Guarro J, Gené J (1995) Opportunistic fusarial infections in humans.

Eur J Clin Microbiol Infect Dis 14:741–754

Guo F, Li Y, Zhao Y, Yi J, Huang C (2017) Dihydroquinolinone

compound with PPAR multiple agonist activity, preparation and

Medicinal Chemistry Research

application for preventing and/or treating metabolic syndrome.

Patent Nº CN 107522657 A 20171229

Gutiérrez RU, Correa HC, Bautista R, Vargas JL, Jerezano AV, Del-

gado F, Tamariz J (2013) Regioselective synthesis of 1,2-dihy-

droquinolines by a solvent-free MgBr₂-catalyzed multicomponent

reaction. J Org Chem 78:9614–9626

Haines BE, Wiest O, Stauffacher CV (2013) The increasingly complex

mechanism of HMG-CoA reductase. Acc Chem Res

46:2416–2426

Harada-Shiba M, Tajima S, Yamamoto A (1995) Response of 3-

hydroxy-3-methylglutaryl CoA reductase to l-triiodothyronine in

cultured ﬁbroblasts from FH homozygotes. Atherosclerosis

113:91–98

Lalloyer F, Staels B (2010) Fibrates, glitazones, and peroxisome

proliferator-activated receptors. Arterioscler Thromb Vasc Biol

30:894–899

La Regina G, D’Auria FD, Taﬁ A, Piscitelli F, Olla S, Caporuscio F,

Nencioni L, Cirilli R, La Torre F, Rodrigues De Melo N, Kelly

SL, Lamb DC, Artico M, Botta M, Palamara AT, Silvestri R

(2008) 1-[(3-Aryloxy-3-aryl)propyl]-1H-imidazoles, new imida-

zoles with potent activity against Candida albicans and derma-

tophytes. Synthesis, structure-activity relationship, and molecular

modeling studies J Med Chem 51:3481–3855

Lee JY, Lee JY, Yun B-S, Hwang BK (2004) Antifungal activity of

β-asarone from rhizomes Acorus gramineus. J Agric Food Chem

52:776–780

Hegedüs A, Hell Z, Vargadi T, Potor A, Gresits I (2007) A new,

simple synthesis of 1,2-dihydroquinolines via cyclocondensation

using zeolite catalyst. Catal Lett 117:99–101

Levine S, Saltzman A (2007) A procedure for inducing sustained

hyperlipemia in rats by administration of a surfactant. J Pharm

Toxicol Methods 55:224–226

Hodel C (2002) Myopathy and rhabdomyolysis with lipid-lowering

drugs. Toxicol Lett 128:159–168

Liu L, Yeh Y-Y (2002) S-Alk(en)yl cysteines of garlic inhibit cho-

lesterol synthesis by deactivating HMG-CoA reductase in cul-

tured rat hepatocytes. J Nutr 132:1129–1134

Man RYK, Lynn EG, Cheung F, Tsang PSY, O K (2002) Cholestin

inhibits cholesterol synthesis and secretion in hepatic cells

(HepG2). Mol Cell Biochem 233:153–158

Huang D, Ou B, Prior RL (2005) The chemistry behind antioxidant

capacity assays. J Agric Food Chem 53:1841–1856

Inoue T, Kinoshita M, Oyama K, Kamemura N, Oyama Y (2018)

Captan-induced increase in the concentrations of intracellular Ca²⁺

and Zn²⁺and its correlation with oxidative stress in rat thymic

lymphocytes. Environ Toxicol Pharm 63:78–83

Maron DJ, Fazio S, Linton MF (2000) Current perspectives of statins.

Circulation 101:207–213

Irwin JJ, Shoichet BK (2005) ZINC–A free database of commercially

available compounds for virtual screening. J Chem Inf Model

45:177–182

Istvan ES, Deisenhofer J (2001) Structural mechanism for statin

inhibition of HMG-CoA reductase. Science 292:1160–1164

Johnson JV, Rauckman BS, Baccanari DP, Roth B (1989) 2,4-Dia-

mino-5-benzylpyrimidines and analogues as antibacterial agentes.

12. 1,2-Dihydroquinolylmethyl analogues with high activity and

speciﬁcity for bacterial dihydrofolate reductase. J Med Chem

32:1942–1949

Matsuda M, Mori T, Kawashima K, Nagatsuka M, Kobayashi S,

Yamamoto M, Kato M, Takai M, Oda T (2007) Preparation of

novel 1,2-dihydroquinoline derivatives having glucocorticoid

receptor binding activity. PCT Int Appl WO 2007032556:A1

20070322

Medina-Franco JL, López-Vallejo F, Rodríguez-Morales S, Castillo R,

Chamorro G, Tamariz J (2005) Molecular docking of the highly

hypolipidemic agent α-asarone with the catalytic portion of

HMG-Co-A reductase. Bioorg Med Chem Lett 15:989–994

Mehta PK, Wei J, Wenger NK (2014) Ischemic heart disease in

Jover-Fernández A, Hernández-Mijares A(2012) Fibratos: efectos

farmacológicos. Clín Inv Arterioscler 24(S1):19–23

Joy TR, Hegele RA (2009) Narrative review: statin-related myopathy.

Ann Intern Med 150:858–868

Kiss LE, Ferreira HS, Torrão L, Bonifácio MJ, Palma PN, Soares-da-

Silva P, Learmonth DA (2010) Discovery of a long-acting, per-

ipherally selective inhibitor of catechol-O-methyltrasferase. J

Med Chem 53:3396–3411

Korolenko TA, Tuzikov FV, Vasil’eva ED, Cherkanova MS, Tuzi-

kova NA (2010) Fractional composition of blood serum lipo-

proteins in mice and rats with Triton WR 1339-induced lipemia.

Bull Exp Biol Med 149:567–570

Kourounakis AP, Victoratos P, Peroulis N, Stefanou N, Yiangou M,

Hadjipetrou L, Kourounakis PN (2002) Experimental hyperlipi-

demia and the effect of NSAIDs. Exp Mol Pathol 73:135–138

Kuroda M, Tanzawa K, Tsujita Y, Endo A (1977) Mechanism for

elevation of hepatic cholesterol synthesis and serum cholesterol

levels in Triton WR-1339-induced hyperlipidemia. Biochim

Biophys Acta 489:119–125

Labarrios F, Garduño L, Vidal MR, García R, Salazar M, Martínez E,

Díaz F, Chamorro G, Tamariz J (1999) Synthesis and hypolipi-

daemic evaluation of a series of α-asarone analogues related to

cloﬁbrate in mice. J Pharm Pharm 51:1–7

Lagu B, Lebedev R, Pio B, Yang M, Pelton PD (2007) Dihydro-[1H]-

quinolin-2-ones as retinoid X receptor (RXR) agonists for

potential treatment of dyslipidemia. Bioorg Med Chem Lett

17:3491–3496

Lalitha MK (2004) Manual on antimicrobial susceptibility testing

(under the auspices of Indian Association of Medical Micro-

biologists). https://www.biodiamed.gr/wp-content/uploads/2017/

06/Manual_on_Antimicrobial_Susceptibility_Testing.pdf Acces-

sed 21 Sept 2004

women:

24:140–151

a focus or risk factor. Trends Cardiovasc Med

Mendieta A, Jiménez F, Garduño-Siciliano L, Mojica-Villegas A,

Rosales-Acosta B, Villa-Tanaca L, Chamorro-Cevallos G,

Medina-Franco JL, Meurice N, Gutiérrez RU, Montiel LE, Cruz

MC, Tamariz J (2014) Synthesis and highly potent hypolipidemic

activity of alpha-asarone- and ﬁbrate-based 2-acyl and 2-alkyl

phenols as HMG-CoA reductase inhibitors. Bioorg Med Chem

22:5871–5882

Menéndez R, Amor AM, Rodeiro I, González RM, González PC,

Alfonso JL, Más R (2001) Policosanol modulates HMG-CoA

reductase activity in cultured ﬁbroblasts. Arch Med Res

32:8–12

Miyazaki A, Koieyama T, Shimada Y, Kikuchi T, Ito K, Kasanuki N,

Koga T (2004) Pravastatin sodium, an inhibitor of HMG-CoA

reductase, decreases HDL cholesterol by transfer of cholesteryl

ester from HDL to VLDL in Japanese white rabbits. J Atheroscler

Thromb 11:22–28

Mohammadzadeh A, Farhat AS, Esmaeli H, Amiri R (2013) Effect of

cloﬁbrate on serum triglyceride and cholesterol after intravenous

lipid in very low birth weight neonates. Iran J Neonatol 4:20–25

Momin RA, Nair MG (2002) Pest-managing efﬁcacy of trans-asarone

isolated from Daucus carota L. seeds. J Agric Food Chem

50:4475–4478

Moreno-Aliaga MJ, Arenas-Vidal JC, Berjón A, Fernández-Otero MP

(1999) Effects of in vivo captan administration on cytotoxicity,

gluconeogenesis, ATP levels, and parameters related to oxidative

stress in rat liver. Pest Biochem Physiol 64:185–193

Morris GM, Huey R, Lindstrom W, Sanner MF, Belew RK, Goodsell

DS, Olson AJ (2009) AutoDock4 and AutoDockTools4: auto-

mated docking with selective receptor ﬂexiblity. J Comput Chem

30:2785–2791

Medicinal Chemistry Research

Motulsky HJ (1999) Analyzing data with GraphPad Prism. GraphPad

Prism Sofware Inc., San Diego, CA. www.graphpad.com

Okopień B, Bułdak Ł, Bołdys A (2018) Beneﬁts and risks of the

treatment with ﬁbrates—a comprehensive summary. Expert Rev

Clin Pharm 11:1099–1112

response via a PPRE in the lipoprotein lipase gene. EMBO J

15:5336–5348

Shaheen B, Barathi K, Prasad KVSRG (2016) Mini review on ther-

apeutic proﬁle of phenoxy acids and their derivatives. Int J Pharm

Pharm Sci 8:66–71

Olender D, Żwawiak J, Zaprutko L (2018) Multidirectional efﬁcacy of

biologically active nitro compounds included in medicines.

Pharmaceuticals 11:54

Shaikh SR, Ali SA (2018) Pleiotropic actions of PPAR agonist: a

potential therapeutic perspectives in the treatment of diabetic

nephropathy. Innov Pharm Pharmacoth 6:6–12

Olivares CR, Alfaro LLJ, Diaz JMC, Thompson ML (2005) Dis-

seminated fusariosis by Fusarium oxysporum in an adult patient

with acute myeloid leukemia and severe febrile neutropenia. Rev

Chil Infect 22:356–360

Oliver M (2012) The cloﬁbrate saga: a retrospective commentary. Br J

Clin Pharm 74:907–910

Pages N, Maurois P, Delplanque B, Bac P, Stables JP, Tamariz J,

Chamorro G, Vamecq J (2010) Activities of α-asarone in various

animal seizure models and in biochemical assays might be

essentially accounted for by antioxidant properties. Neurosci Res

68:337–344

Silva RM, Santos FA, Maciel MAM, Pinto AC, Rao VSN (2001)

Effect of trans-dehydrocrotonin, a 19-nor-clerodane diterpene

from Croton cajucara on experimental hypertriglyceridaemia and

hypercholesterolaemia induced by Triton WR 1339 (tyloxapol) in

mice. Planta Med 67:763–765

Singh N, Tamariz J, Chamorro G, Medina-Franco JL (2009) Inhibitors

of HMG-CoA reductase: current and future prospects. Mini-Rev

Med Chem 9:1272–1283

Steinberg D (2005) Hypercholesterolemia and inﬂammation in ather-

ogenesis: two sides of the same coin. Molec Nutr Food Res

49:995–998

Panini SR, Sexton RC, Gupta AK, Parish EJ, Chitrakorn S, Rudney H

(1986) Regulation of 3-hydroxy-3-methylglutaryl coenzyme A

reductase activity and cholesterol biosynthesis by oxylanosterols.

J Lipid Res 27:1190

Parker RA, Pearce BC, Clark RW, Gordon DA, Wright JJK (1993)

Tocotrienols regulate cholesterol production in mammalian cells

by post-transcriptional suppression of 3-hydroxy-3-methylglu-

taryl-coenzyme A reductase. J Biol Chem 268:11230–11238

Pfaller MA, Diekema DJ (2012) Progress in antifungal susceptibility

testing of Candida spp. by use of Clinical and Laboratory Stan-

dards Institute broth microdilution methods, 2010 to 2012. J Clin

Microbiol 50:2846–2856

Sung JH, Lee S-J, Park KH, Moon TW (2004) Isoﬂavones inhibit 3-

hydroxy-3-methylglutaryl coenzyme A reductase in vitro. Biosci

Biotechnol Biochem 68:428–432

Thagizadeh E, Esfehani RJ, Sahebkar A, Parizadeh SM, Rostami D,

Mirinezhad M, Poursheikhani A, Mobarhan MG, Pasdar A

(2019) Familial combined hyperlipidemia: an overview of the

underlying molecular mechanisms and therapeutic strategies.

IUBMB Life. https://doi.org/10.1002/iub.2073

The Field Study Investigators (2005) Effects of long-term fenoﬁbrate

therapy on cardiovascular events in 9795 people with type 2

diabetes mellitus (the FIELD study): randomised controlled trial.

Lancet 366:1849–1861

Pisoschi AM, Pop A (2015) The role of antioxidants in the chemistry

of oxidative stress: a review. Eur J Med Chem 97:55–74

Poplawsky J, Lozowicka B, Dubis AT, Lachowska B, Witkowski S,

Siluk D, Petrusewicz J, Kaliszan R, Cybulski J, Strzalkowska M,

Chilmonczyk Z (2000) Synthesis and hypolipidemic and anti-

platelet activities of α-asarone isomers in humans (in vitro), mice

(in vivo), and rats (in vivo). J Med Chem 43:3671–3676

Qiao J, Kontoyiannis DP, Wan Z, Li R, Liu W (2007) Antifungal

activity of statins against Aspergillus species. Med Mycol

45:589–593

Raju J, Bird RP (2007) Diosgenin, a naturally occurring furostanol

saponin suppresses 3-hydroxy-3-methylglutaryl CoA reductase

expression and induces apoptosis in HCT-116 human colon

carcinoma cells. Cancer Lett 255:194–204

Ramírez-Villalva A, González-Calderón D, Rojas-García RI, Gonzá-

lez-Romero C, Tamaríz-Mascarúa J, Morales-Rodríguez M,

Zavala-Segovia N, Fuentes-Benítes A (2017) Synthesis and

antifungal activity of novel oxazolidin-2-one-linked 1,2,3-triazole

derivatives. Med Chem Commun 8:2258–2262

Trzaskos JM, Magolda RL, Favata MF, Fischer RT, Johnson PR, Chen

HW, Ko SS, Leonard DA, Gaylor JL (1993) Modulation of 3-

hydroxy-3-methylglutaryl-CoA reductase by 15α-ﬂuorolanost-7-

en-3β-ol. J Biol Chem 268:22591–22599

Türkkan M, Erper I (2015) Inhibitory inﬂuence of organic and inor-

ganic sodium salts and synthetic fungicides against bean root rot

pathogens. Gesund Pﬂanz 67:83–94

Uchida A, Slipchenko MN, Cheng J-X, Buhman KK (2011) Fenoﬁ-

brate, a peroxisome proliferator-activated receptor α agonist,

alters triglyceride metabolism in enterocytes of mice. Biochim

Biophys Acta 1811:170–176

U.S. Environmental Protection Agency (1984) Health and environ-

ment effects proﬁle for captan. EPA/600/x-84/253. Environ-

mental Criteria and Assessment Ofﬁce, Ofﬁce of Health and

Environmental Assessment, Ofﬁce of Research and Develop-

ment, Cincinnati, OH

Van Gaal LF, Mertens IL, De Block CE (2006) Mechanisms linking

obesity with cardiovascular disease. Nature 444:875–880

van Meerloo J, Kaspers GJ, Cloos J (2011) Cells sensitivity assays: the

MTT assay. Methods Mol Biol 731:237–245

Veseli BE, Perrotta P, De Meyer GRA, Roth L, Van der Donckt C,

Martinet W, De Meyer GRY (2017) Animal models of athero-

sclerosis. Eur J Pharm 816:3–13

Vivancos J, León-Colombo T, Monforte-Dupret C (1999) Hypolipi-

demic treatment in the prevention of atherosclerotic plaque

complications. Rev Neurol 29:857–863

Rashid M, Khan R, Khan RA (2018) Microwave assisted catalytic

region-selective nitration of resorcinol and substituted phenols.

Chem Res J 3:17–24

Rodríguez-Páez L, Juárez-Sánchez M, Antúnez-Solís J, Baeza I, Wong

C (2003) α-Asarone inhibits HMG-CoA reductase, lowers serum

LDL-cholesterol levels and reduces biliary CSI in hypercholes-

terolemic rats. Phytomedicine 10:397–404

Rotta-Bonﬁn M, Bulle-Oliveira AS, do Amaral SL, Luiz-Monteiro H

(2015) Treatment of dyslipidemia with statins and physical

exercises: recent ﬁndings of skeletal muscle responses. Arq Bras

Cardiol 104:324–332

Wierzbicki AS (2001) Atorvastatin. Expert Opin Pharmacother 2:819–830

Williams D, Feely J (2002) Pharmacokinetic–pharmacodynamic drug

interactions with HMG-CoA reductase inhibitors. Clin Pharma-

cokinet 41:343–370

SAS Institute Inc (9.0) (2014) SAS/ACCESS® 9.0, Cary: NC.

https://www.sas.com/search/en_us.html?q =SAS/ACCESS%209.0

Schoonjans K, Peinado-Onsurbe J, Lefebvre A-M, Heyman RA,

Briggs M, Deeb S, Staels B, Auwerx J (1996) PPARα and

PPARγ activators direct a distinct tissue-speciﬁc transcriptional

Willson TM, Brown PJ, Sternbach DD, Henke BR (2000) The PPARs:

from orphan receptors to drug discovery. J Med Chem 43:527–550

Xiao Z, Fei L, Chen X, Wang X, Cao L, Ye K, Zhu W, Xu S (2018)

Design, synthesis, and antitumor evaluation of quinoline-

imidazole derivates. Arch Pharm Chem Life Sci 351:e1700407

Medicinal Chemistry Research

Yuan G, Al-Shali KZ, Hegele RA (2007) Hypertriglyceridemia: its

etiology, effects and treatment. Can Med Assoc J 176:1113–1120

Zeniya M, Reuben A (1988) Triton WR-1339-induced changes in serum

lipids and biliary lipid secretion. Am J Physiol 254:G346–G354

Zhang Y, Fang Y, Liang H, Wang H, Hu K, Liu X, Yi X, Peng Y

(2013) Synthesis and antioxidant activities of 2-oxo-quinoline-3-

carbaldehyde Schiff-base derivatives. Bioorg Med Chem Lett

23:107–111

Zúñiga C, Garduño L, Cruz MC, Salazar M, Pérez-Pastén R, Cha-

morro G, Labarrios F, Tamariz J (2005) Design of new potent

hypolipidemic agents with the synergistic structural properties of

α-asarone and ﬁbrates. Drug Dev Res 64:28–40

Article Doi

Synthesis and biological activity of fibrate-based acyl- and alkyl-phenoxyacetic methyl esters and 1,2-dihydroquinolines

DOI: 10.1007/s00044-019-02496-1

Source and publish data:

Authors:

Article abstract of DOI:10.1007/s00044-019-02496-1

Full text of DOI:10.1007/s00044-019-02496-1

Products guided by the article

R&D Labs maybe for 1246085-12-0

Relevant to this article

Hot Product