Synthesis, functionalization and biological activity of arylated derivatives of (+)-estrone

Accepted Manuscript

Synthesis, Functionalization and Biological Activity of Arylated Derivatives of

(+)-Estrone

Anton Ivanov, Syeda Abida Ejaz, Syed Jawad Ali Shah, Peter Ehlers, Alexander

Villinger, Eva Frank, Gyula Schneider, János Wölfling, Qamar Rahman,

Jamshed Iqbal, Peter Langer

PII:

S0968-0896(16)31388-8

DOI:

http://dx.doi.org/10.1016/j.bmc.2016.12.009

BMC 13436

Reference:

Bioorganic & Medicinal Chemistry

To appear in:

Received Date:

Revised Date:

Accepted Date:

12 September 2016

6 December 2016

7 December 2016

Please cite this article as: Ivanov, A., Abida Ejaz, S., Jawad Ali Shah, S., Ehlers, P., Villinger, A., Frank, E.,

Schneider, G., Wölfling, J., Rahman, Q., Iqbal, J., Langer, P., Synthesis, Functionalization and Biological Activity

of Arylated Derivatives of (+)-Estrone, Bioorganic & Medicinal Chemistry (2016), doi: http://dx.doi.org/10.1016/

j.bmc.2016.12.009

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1

Synthesis, Functionalization and Biological Activity of Arylated

Derivatives of (+)-Estrone

Anton Ivanov,^aSyeda Abida Ejaz,^dSyed Jawad Ali Shah,^dPeter Ehlers,^a,bAlexander Villinger,^aEva

Frank,^cGyula Schneider,^cJános Wölfling,^cQamar Rahman,^eJamshed Iqbal,^dPeter Langer^a,b

*

a

Institut für Chemie, Universität Rostock, Albert Einstein Str. 3a, 18059 Rostock, Germany

^bLeibniz-Institut für Katalyse an der Universität Rostock e.V., Albert Einstein Str. 29a, 18059 Rostock,

Germany; peter.langer@uni-rostock.de

^cDepartment of Organic Chemistry, University of Szeged, Dóm tér 8, H-6720 Szeged, Hungary

^dCenter for Advanced Drug Research, COMSATS Institute of Information Technology,

Abbottabad-22060, Pakistan

^eAmity University, Lucknow Campus, Viraj Khand-5, Gomti Nagar, Lucknow - 226010, India

Abstract.

Various novel arylated estrone derivatives, such as 2-aryl-, 4-aryl- and 2,4-diaryl-estrones, by Suzuki-

Miyaura reactions. While the synthesis of 4-arylestrones could be carried out under standard conditions,

the synthesis of 2-arylestrones and 2,4-diarylestrones required a thorough optimization of the conditions

and it proved to be important to use sterically encumbered biaryl ligands. The best results were obtained

by the use of RuPhos. Combination of developed Suzuki coupling reactions with subsequent cyclization

reactions afforded more complex hybrid structures, containing dibenzofuran, benzocoumarin and steroid

moieties. These derivatives were tested as pancreatic lipase inhibitors and it was found that most of the

compounds exhibited inhibition of pancreatic lipase but the maximum inhibitory potential was shown by

4-arylestrones. All of the synthesized derivatives showed inhibitory values in the range of 0.82 0.01 to

59.7 3.12 µM. The biological activity was also rationalized on the bases of docking studies.

Graphical abstract

2

Introduction

Obesity is the fifth leading cause of death in the world and has gained much attention because of its role

in different types of cancers, including breast (postmenopausal), pancreatic, gallbladder and liver

cancer.^1aThe exact biological mechanisms relating obesity and cancer are complex and still not clearly

defined. However, these mechanism may include obesity-related hormones especially estrogen, growth

factors, modulation of energy balance and calorie restriction affecting the cancer cell promotion and

growth.^1bTherefore, control of obesity can minimize the risk of cancer. Within the human body lipid

metabolism is controlled by the group of enzymes known as lipases.²This group of enzymes includes

pancreatic and gastric lipases, which are responsible for the hydrolysis of triacylglycerols into free fatty

acids.³In muscles, these free fatty acids act as a source of energy, whereas in adipose tissue they undergo

re-esterification and are ultimately stored. In comparison to gastric lipase, pancreatic lipase is mainly

involved in the digestion of dietary fats up to 50-70%.⁴Thus, dietary fat is an important factor that

contributes to hyperlipidemia and associated to a variety of diseases including obesity, hypertension,

diabetes and cardiovascular problems.⁵It means the inhibition of lipases can help in limiting the

hydrolysis of dietary fats which will in turn help to control the obesity.^6aAs a result, further complications

arising from obesity can also be prevented.

6c

Different synthetic derivatives including phenethylamine (sibutramine),^6bcarboxamide (rimonabant)

and lipstatin (orlistat) ^6care avialable on the market and are being used as anti-obesity drugs. Sibutramine

is a centrally acting drug which causes the inhibition of reuptake of nor-adrenaline, serotonin and

dopamine.^6bWhilst, rimonabant acts through the inhibition of cannabinoid-1 receptors. Although being

effective antiobesity drugs, both of these drugs are associated with several side effects including

insomnia, headache, hypertension, palpitation, dizziness, constipation and anxiety disorders.^6b,c

3

1

2

3

Figure 1: Currently avvailable anti-obesity drug (1) Sibutramine (2) rimonabant (3) orlistat.^6b-c

Among above mentioned derivatives, only orlistat is considered to be acting directly through inhibition of

pancreatic lipase and is able to prevent absorption of approximately 30% of dietary fat.^7aApart from its

antiobesity activity, it can also modestly decrease blood pressure, prohibit the onset of diabetes type-2 and

improve oral glucose tolerance. The mechanism of lipase inhibition by orlistat is via its covalent bonding

to the serine which is present in the active site of the lipase.^7bDespite the promising results of orlistat for

obesity treatment, it is associated with certain unpleasant gastrointestinal side effects. The most important

adverse effects are diarrhea, liquid stools, fecal urgency, flatulence, oily spotting and abdominal

cramping.⁸Due to the adverse effects of orlistat, it may not be well tolerated. Many derivative of

alkaloids,^9aglycosides,^9bcarotinoids,^9csaponins,^9dterpenes^9eand polyphenoles^9fhave also been isolated

from plants and reported as pancreatic lipase inhibitors (Figure 2). Hence, it is crucial to identify different

compounds (from natural and synthetic source) to overcome obesity and its associated complications.

Therefore more researches have been focused on the identification of newer pancreatic lipase inhibitors

with less unpleasant adverse effects.

(IC₅₀= 15.83 1.10 µM)

(IC₅₀= 12 µg/mL)

(IC₅₀= 41.8 µM)

(IC₅₀>20 µM)

1

2

3

4

Figure 2: Previously reported pancreatic lipase inhibitors (1) ursolic acid (2) carnosic acid (3) cassiamin

A (4) (-)-epicatechin.^9e,f

Within the human body many steroidal molecules are playing specific physiological functions. For

example, cholesterol is an essential structural component of all animal cell membranes, whereas steroid

hormones are important part of a signal system within the organisms.¹⁰Among the steroidal hormones,

female primary sex hormones (estrogens) promote the development of female secondary sexual

characteristics and regulate the menstrual cycle.^11a(+)-Estrone (1) in Figure 3, is an interesting example of

such steroidal molecules. ^11bDifferent derivatives of estrones have been reported as anticancer agents and

used as component of oral contraceptive pills.¹²Moreover, different studies have been reported on 2- and

4-substituted estrones e.g. 1-adamantyl compound ZYC-26 (2) as neuroprotective molecule against

4

cerebralischemia/reperfusion injury^13a,band 2- and 4-substituted estrones (3) derivatives as steroid

sulfatase (STS) inhibitors (Figure 3).^14a-cThere are, however, no studies concerning the effects of the

presence of aryl groups located at positions 2 and/or 4 on the bioactivity of steroids.

O

1

R

2

HO

2

4

R

Me

, ZYC-26

1

, R = H, F, CN, NO ;

2

3

2

1

, (+)-Estrone

2

R = H, NO

2

Figure 3. Estrone and its biologically active derivatives^12-14

As mentioned before, 2- and/or 4-aryl-substituted estrones have only been scarcely studied and there are

only a few preparative methods towards these compounds in the literature focusing on transition metal

catalyzed CH-activation.^{15a, 11b}However, these methods suffer from either moderate yields, requirement

of a directing group or the restriction of special nucleophiles. Moreover, non-selective arylation by

stoichiometric organo-bismuth derivatives are known to give access to the target products in low

yields.^16a,bThus, we decided to investigate the scope and limitations of the synthesis of arylated estrone

derivatives. On the basis of previous studies,^9e,fit was observed that steroid derivatives isolated from

plants inhibited the pancreatic lipase, therefore, we tested our compounds as lipase inhibitors.

Results and Discussions

The Suzuki-Miyaura cross-coupling reaction is a convenient and powerful tool for diaryl synthesis and

the reaction is characterized by a good functional group tolerance. Therefore, we decided to apply this

reaction using 2- and 4-bromoestrone and 2,4-dibromoestrone as starting materials. All substrates were

easily synthesized by simple bromination¹⁷using either N-bromosuccinimide or 1,3-dibromo-5,5-

dimethylhydantoin starting from estrone (Scheme 1).

Scheme 1. Synthesis of substrates 6 a-c. Conditions: i, 1 (1.0 equiv.), N-bromosuccinimide (1.1 equiv.),

CCl₄, reflux, 3h; ii, 1 (1.0 equiv.), N-bromosuccinimide (1.1 equiv.), DCM, 20°C, 1h; iii, 1 (1.0 equiv.),

1,3-dibromo-5,5-dimethylhydantoin (1.1 equiv.), DCM, 20 °C, 1h.

5

4-Arylestrones. As a first step we decided to study the Suzuki-Miyaura reaction of 4-bromoestrone. To

our delight, application of standard reaction conditions, using 5 mol% of Pd (PPh₃)₄, gave the desired 4-

arylestrones 7 a-m in good to excellent yields (Scheme 2, Table 1). The application of boronic acids

containing electron-donating as well as electron-withdrawing groups gave equally good results, while

more sterically hindered o-substituted boronic acids (7 k-m) resulted in lower yields. Reduction of the

amount of catalyst to 2.5 mol% resulted in a slightly reduced yield. Using DMF instead of dioxane

resulted in a dramatically decreased yield.

Scheme 2. Synthesis of 4-arylestrones 7 a-m. Conditions: i, 6 a (1.0 equiv.), boronic acid (1.5 equiv.),

Pd(PPh₃)₄(5 mol%), K₃PO₄(2.0 equiv.), dioxane, 101°C, 8 h

Table 1. Synthesis of 4-arylestrones 7 a-m.

7

Ar

C₆H₅

7 [%]^a

88

A

B

4-MeC₆H₄

98

C

4-t-BuC₆H₄

4-(MeO)C₆H₄

4-(F₃C)C₆H₄

4-(F₃CO)C₆H₄

3-FC₆H₄

90

D

99

E

91

F

96

G

92

H

3,5-(MeO)₂C₆H₃

4-EtC₆H₄

97

I

99

J

4-ClC₆H₄

87

K

2-FC₆H₄

94

L

M

2,3,4-(MeO)₃C₆H₂

2-(EtO)C₆H₄

67

79

^aIsolated yields.

It is worth to be noted that, in case of o-substituted products 7 k-m new atropisomers are formed due to

hindered rotation, thus forming a mixture of two diastereomers. This was proven by the observation of

two sets of NMR signals. Interestingly, even small atoms, such as Fluor, were sufficient for the

6

occurrence of the described effect (Figure 4). The structure of 7 c was independently confirmed by X-ray

analysis (Figure 5).

Figure 4. M- and P-Atropisomers of 7 k.

Figure 5. Molecular structure of 7c (Ortep plot showing thermal ellipsoids at the 50% probability level).

2-Arylestrones. The reaction of 2-bromoestrone (6 b) with p-tolylboronic acid was next studied (Table

2). We first tested the application of the same conditions as applied for 4-bromoestrone. However, the

reactions were unsuccessful and resulted in the formation of complex mixtures. Afterwards, we applied

different catalysts, ligands, bases and solvents to find appropriate conditions. Thus, we found that the use

of Buchwald’s ligands RuPhos and SPhos were crucial to obtain desired arylated product. The use of

RuPhos gave the best yields (Table 2). The requirement of such sophisticated catalytic systems was

somehow surprising as transition metal catalysts exclusively gave 2-arylated instead of 4-arylated

15a, 11b

products in CH-activation reactions.

In particular, Bedford et al. described CH-arylation of

carbamate protected estrones with aryliodides which were mainly governed by steric effects. However, in

our case the cross-coupling reaction might be effected primarily by electronic effects as the sterically less

hindered position 2 is only addressed by a sophisticated catalyst system.

7

Table 2. Optimization of the Suzuki reaction for 2-tolylestrone 8 b.

i

Conditions: 6 b (1.0 equiv.), p-tolylboronic acid (1.5 equiv.), catalyst (5.0 mol%), ligand (5.0 mol%),

base (2.0 equiv.), solvent.

Entry

1

Catalyst

Pd(PPh₃)₄

Ligand

-

Base

Solvent T [^oC] Time [h] Yield [%]

b

K₃PO₄

dioxane

101

130

101

105

101

20

18

20

22

28

20

-

b

2

-

Cs₂CO₃dioxane

3

-

K₃PO₄

K₂CO₃

K₃PO₄

DMF

4

BINAP

SPhos

XPhos

RuPhos

PCy₃·HBF₄

RuPhos

PCy₃·HBF₄

-

DMF

5

dioxane

66^a

b

6

-

7

75^a

35^a

69^a

40^a

63^a

73^a

8

9

10

11

12

13

14

15

Cs₂CO₃dioxane

KF

dioxane

toluene

DMF

K₂CO₃

c

-

d

b

Pd(OAc)₂

dioxane

-

b

PdCl₂(PPh₃)₂

dioxane

-

^aIsolated yields. ^bLow conversion of starting material. ^cInseparable mixture. ^d10 mol%.

With this knowledge in hand, we obtained 2-arylestrones 8 a-h in good yields. As expected, all yields

were significantly lower in comparison with the corresponding yields of 4-arylestrones, due to the lower

reactivity of this position (Table 3). Electronic properties of the substituents were not affecting the yield.

Application of o-sustituted arylboronic acids as well as p-chlorophenylboronic acid was unsuccessful.

The latter can be explained by the occurrence of the competing cross-coupling reaction between two

8

molecules of arylboronic acids. The structures of 8 c and 8 e were independently confirmed by X-ray

crystal structure analyses (Figures 6 and 7).

Table 3. Synthesis of 2-arylestrones 8 a-k.

Conditions: i, 6 b (1.0 equiv.), boronic acid (1.5 equiv.), Pd(PPh₃)₄(5 mol%), RuPhos (5 mol%), K₂CO₃

(2.0 equiv.), dioxane, 101°C, 16 h.

8

A

B

C

D

E

F

G

H

I

Ar

8 [%]^a

75

C₆H₅

4-MeC₆H₄

75

4-t-BuC₆H₄

4-(MeO)C₆H₄

4-(F₃C)C₆H₄

4-(F₃CO)C₆H₄

3-FC₆H₄

82

80

72

77

79

3,5-(MeO)₂C₆H₃

2-FC₆H₄

84

b

-

b

J

2,3,4-(MeO)₃C₆H₂

4-ClC₆H₄

-

b

K

-

^aIsolated yields. ^bInseparable mixture.

9

Figure 6. Molecular structure of 8 (Ortep plot showing thermal ellipsoids at the 50% probability level). A

solvent molecule (methanol) has been omitted for clarity.

Figure 7. Molecular structure of 8 e (Ortep plot showing thermal ellipsoids at the 50% probability level).

The disordered atoms of the CF₃group have been omitted for clarity.2,4-Diarylestrones. The conditions

used for the synthesis of 2-bromoestrone (6 b) were successfully applied to 2,4-dibromoestrone (6 c). In

contrast, the application of milder conditions used for 4-bromoestrone (6 a) resulted in the formation of

an inseparable mixture of starting material, di- and mono-substituted products. To exclude formation of

the latter we prolonged the reaction time (40 h), applied higher temperature (110 °C) and, hence, used

toluene as the solvent. The reactions provided the pure disubstituted products 9 a-o in excellent yields

(Table 4). The electronic properties of the arylboronic acids did not affect the yields, while sterically

hindered o-arylboronic acids surprisingly gave the corresponding products in good yields. In general, in

comparison with the 2-arylestrones, the products were obtained in higher yields. That fact makes us

assume that position 2 is strongly affected by the presence of either a bromine atom or an arylring located

at position 4, which influences the reactivity of this position and facilitates the reaction. The application

of only one equivalent of arylboronic acid gave an inseparable mixture of starting material, mono- and di-

arylated products, which underlines the higher reactivity of 6 c or an 4-arylated intermediated compared

to 6 b. Similar to 4-arylestrones, ortho-substituted products 9 j, 9 m and 9 n exist as a mixture of two

diastereomers. Interestingly, the presence of ortho-substituted aryl groups at position 2 did not result in

formation of an additional stereogenic axis, which can be observed by NMR spectroscopy.

Table 4. Synthesis of 2,4-arylestrones 9 a-o.

10

Conditions: i, 6 c (1.0 equiv.), boronic acid (3 equiv.), Pd(PPh₃)₄(5 mol%), RuPhos (5 mol%), K₂CO₃

(4.0 equiv.), toluene, 110 °C, 40 h.

9

A

B

C

D

E

F

Ar

C₆H₅

9 [%]^a

99

4-MeC₆H₄

4-t-BuC₆H₄

4-(MeO)C₆H₄

4-(F₃C)C₆H₄

4-(F₃CO)C₆H₄

3-FC₆H₄

91

85

99

89

79

G

H

I

85

3,5-(MeO)₂C₆H₃

4-EtC₆H₄

70

99

J

2-FC₆H₄

97^b

K

L

M

N

O

3,5-Me₂C₆H₃

4-ClC₆H₄

99

24

36^b

2,3,4-(MeO)₃C₆H₂

2-(EtO)C₆H₄

4-FC₆H₄

72

74

^aIsolated yields. ^bReaction time: 72 h.

Further functionalization. We next synthesized, based on the Suzuki reactions described above, novel

polycyclic ring structures which contain a steroid and a dibenzofuran moiety. Treatment of 7 k with

K₂CO₃in DMF at 150 °C afforded the desired product 10 in 69% yield (Scheme 3). Likewise, product 11

was synthesized starting with 9 j. Fortunately, cyclized product 11 was obtained as the only product and

no other isomer, which might derive from cyclization of the 2-fluorophenyl moiety was observed. The

structure of 11 was unambiguously confirmed by NMR spectroscopy. During the optimization, we

modified a protocol mentioned in a patent¹⁸for a related transformation and used DMF instead of N-

methylpyrrolidone and applied a lower temperature than that reported. We also tried to prepare 10 and 11

directly from 6 a and 6 c by a one-pot protocol (Suzuki coupling and subsequent base mediated

cyclization). In this case, products 10 and 11 were isolated in 34% and 65% overall yields. Thus, the

stepwise procedure is better in terms of the yield.

11

Scheme 3. Synthesis of cyclized products 10 and 11. Conditions: i, 7 k or 9 j (1.0 equiv.), K₂CO₃(2.0

equiv.), DMF, 150 °C, 4 h.

In a similar manner, we planned to perform a synthesis of lactone 12. Unfortunately, we did not achieve

to prepare the required o-carboxyphenyl derivative of any of the three substrates as an inseparable

mixture of starting material, product and side-products was obtained in all cases. However, the harsh

conditions used for the arylation of 2,4-bromoestrone, applied for 4-bromoestrone, resulted in a direct

one-pot formation of the desired lactone in good yield (Scheme 4). The structure of 12 was independently

confirmed by X-ray crystal structure analysis (Figure 8).

Scheme 4. Synthesis of lactone 12. Conditions: i, 6 a (1.0 equiv.), ortho-ethoxycarbonylphenylboronic

acid (3 equiv.), Pd(PPh₃)₄(5 mol%), RuPhos (5 mol%), K₂CO₃(4.0 equiv.), toluene, 110 °C, 72 h.

12

Figure 8. Molecular structure of 12 (Ortep plot showing thermal ellipsoids at the 50% probability level).

Lipase Inhibition Studies and SAR:

Different derivatives of 4-arylestrones, 2-arylestrones and 2,4-diarylestrones were synthesized and tested

for their inhibitory potential against lipase. Initial screening was performed at the final concentration of

100 µM. Most of the derivatives were found as potent inhibitors of lipase. It was observed that 4-

arylestrones exhibited a higher inhibition of lipase as compared to 2-arylestrones and 2,4-diarylestrones

derivatives. Among the 4-arylestrone derivatives, compound 7 e were found as the most potent inhibitor

having the inhibitory value of IC₅₀SEM = 0.82 0.01 µM. The inhibitory potential of 7 e (4-arylestrone)

was higher than 8 e (the corresponding 2-arylestrone derivative) and 9 e (the corresponding 2,4-

diarylestrone derivative) having the same substituent located at the benzene ring. Table 5 shows, that

substitution of the phenyl ring has substantial effects on the activity. Especially the introduction of

fluorine containing groups (particularly compounds 7 e and 7 g) improved the activity. Interestingly,

compound 7 c (IC₅₀SEM = 1.78 0.05 µM) with donating tBu-group exhibited good inhibitory activity,

too. Except compound 7 h (IC₅₀SEM = 59.7 3.12 µM) and 7 i (IC₅₀SEM = 13.6 0.99 µM), the

inhibitory activities (IC₅₀SEM) were in the range of 0.82 0.01 to 6.42 0.44 µM. Similarly in case of 2-

arylestrones and 2,4-diarylestrones inhibitory activity was substantially effected by the substitution

pattern. 2-Arylestrones were less active compared to their regioisomers 7, except 2-tolylestrone 8 b which

showed good activity (IC₅₀SEM = 1.21 0.02µM). Regarding diarylestrones, compound 9 o showed very

high activity (IC₅₀SEM = 0.82 0.02µM), containing again a fluorine substituent in para-position of the

arylrings. Remarkably, benzofuran derivative 11 performed very well in the studies

(IC₅₀SEM = 0.93 0.04µM), while 9 j with the same substituent in position 2 showed very low activity.

13

Table 5. Porcine Pancreatic lipase inhibition in the presence of the synthesized compounds.

Antilipase

acivity

Antilipase

acivity

Antilipase

acivity

Compound

IC₅₀(µM) SEM/% inhibition

7a

7b

7c

7d

7e

7f

1.54 0.11

38%

8c

8d

8e

8f

5.66 0.76

12.3 0.78

16.3 1.09

2.54 0.11

16.3 1.11

16.4 1.23

25.6 1.67

39%

9i

9j

41%

43%

1.78 0.05

42%

9k

24.1 1.21

1.75 0.07

37%

9l

0.82 0.01

6.42 0.44

1.34 0.11

59.7 3.12

13.6 0.99

45%

8g

8h

9a

9b

9c

9d

9e

9f

9m

9o

0.82 0.02

32%

7g

7h

7i

10

11

0.93 0.04

1.36 0.02

0.31 0.02

8.29 0.66

5.28 0.66

42%

12

7j

Orlistat

7l

43%

8a

8b

39%

1.21 0.03

7.19 0.32

1.21 0.02

9g

The experiment was carried out in triplicate at final conc. of 100 µM and IC₅₀values are expressed as mean SEM . IC₅₀is the

concentration at which 50% of the enzyme activity is inhibited.

Conclusion

In the present study, we have developed conditions for the arylation of bromoestrones by Suzuki-Miyaura

reactions. This method allows the preparation of 2-aryl-, 4-aryl- and 2,4-diaryl-estrones in good to

excellent yields. Further transformations gave access to hybrids of dibenzofuran and benzocoumarin.

Their biological activity studies showed that the synthesized compounds inhibited porcine pancreas lipase

enzyme in up to sub-micromolar concentrations. 4-Arylestrone 7 e and 2,4-diarylestrone 9 o showed the

highest inhibitory activity with an IC₅₀=0.82 0.01 µM, followed by bezofuran-derivative 11 with

IC₅₀SEM = 0.93 0.04µM.

Experimental section

Chemistry

If not otherwise stated NMR data were recorded at room temperature on a Bruker AVANCE 250 MHz,

1

300 MHz or 500 MHz. Chemical shifts in ppm of H and ¹³C NMR spectra were referenced to residual

protons of deuterated solvents CDCl₃(¹H, 7.26 ppm; ¹³C, 77.16 ppm) or DMSO-d6 (¹H, 2,50 ppm; ¹³C,

39.52 ppm). Peak characterization: s = singlet, brs = broad singlet, d = doublet, t = triplet, m = multiplet,

14

dd = doublet of doublets. Infrared spectra were recorded on a Nicolet 550 FT - IR spectrometer with ATR

sampling technique. Gas-chromatography-mass-analysis was carried out on an Agilent HP-5890

instrument with an Agilent HP-5973 Mass Selective Detector (EI) and HP-5 capillary column using

helium carrier gas. ESI HRMS measurements were performed on an Agilent 1969A TOF mass

spectrometer. For High Resolution MS (HRMS) Finnigan MAT 95 XP was used. X-Ray single crystal

structure analysis was performed on a Bruker-Nonius Apex X8 CCD-diffractometer. Data of X-Ray

crystal structure analyses were deposited at the CCDC.¹⁹Melting points were determined on a Micro-Hot-

Stage GalenTM III Cambridge Instruments. The melting points are not corrected.

4-Phenyl-3-hydroxyestra-1,3,5(10)-trien-17-one (7 a):¹⁶4-Bromoestrone (6 a) (100 mg, 0.286 mmol),

boronic acid (1.5 equiv.), Pd(PPh₃)₄(17.0 mg, 0.05 equiv.), K₃PO₄(121.5 mg, 2.0 equiv.) and dioxane (4

ml) were added in a pressure tube under argon atmosphere. The mixture was stirred at 101 °C overnight.

The solvent was evaporated in vacuo. The residue was purified by column chromatography (silica gel,

heptane/EtOAc = 1:5). 7 a was isolated as a light yellow solid (87 mg, 88%), mp = 89

– 90 °C, [α]_D=

+86.4° (c 1.00, CHCl₃) (lit. [α]_D= +83.1° (c 1.77, EtOH)). ¹H NMR (300 MHz, CDCl₃): δ = 0.93 (s, 3H,

CH₃), 1.31 – 1.39 (m, 1H, aliphatic), 1.45 – 1.65 (m, 5H, aliphatic), 1.87 – 2.16 (m, 4H, aliphatic), 2.26 –

3

2.33 (m, 1H, aliphatic), 2.42 – 2.58 (m, 4H, aliphatic), 4.62 (br.s, 1H, OH), 6.86 (d, J_H-H= 8.6 Hz, 1H,

CH_Ar), 7.24 – 7.30 (m, 3H, CH_Ar+ CDCl₃), 7.40 – 7.45 (m, 1H, CH_Ar), 7.48 – 7.455 (m, 2H, CH_Ar). ¹³

C

NMR (75 MHz, CDCl₃): δ = 13.9 (CH₃), 21.6 (CH₂), 26.2 (CH₂), 26.5 (CH₂), 28.5 (CH₂), 31.6 (CH₂),

35.9 (CH₂), 37.8 (CH), 44.4 (CH), 48.0 (C), 50.4 (CH), 112.6 (CH), 125.9 (CH), 127.5 (C), 128.2 (CH),

129.5 (2CH), 130.1 (CH), 130.5 (CH), 132.0 (C), 135.41 (C), 135.45 (C), 150.9 (C), 2.9 (C=O). IR (ATR,

cm⁻¹):

˜ = 3363 (w), 2924 (m), 1733 (s), 1587 (w), 1472 (m), 1166 (m), 762 (m), 701 (s). MS (EI, 70

ν

eV): m/z (%) = 346 (100) [M⁺], 347 (27), 261 (28), 248 (14), 235 (10), 222 (28), 221 (12), 207 (10), 202

(14), 189 (10), 181 (14), 178 (10), 165 (15), 55 (11), 41 (15). HRMS (EI, 70 eV): calcd. for C₂₄H₂₆O₂

[M⁺] 346.19273; found 346.19267.

4-(p-Tolyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (7 b): light yellow solid (102 mg, 99%), mp = 104

–

105 °C, [α]_D= +78.4° (c 1.00, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.93 (s, 3H, CH₃), 1.28 – 1.39

(m, 1H, aliphatic), 1.49 – 1.63 (m, 5H, aliphatic), 1.86 – 2.18 (m, 4H, aliphatic), 2.26 – 2.34 (m, 1H,

aliphatic), 2.49 (s, 3H, CH₃), 2.49 – 2.53 (m, 4H, aliphatic), 4.68 (br.s, 1H, OH), 6.84 (d, ³J_H-H= 8.54 Hz,

1H, CH_Ar), 7.13-7.19 (m, 2H, CH_Ar), 7.23 – 7.26 – (m, 1H, CH_Ar+ CDCl₃), 7.30 – 7.34 (m, 2H, CH_Ar).

¹³C NMR (63 MHz, CDCl₃): δ = 13.8 (CH₃), 21.3 (CH₃), 21.5 (CH₂), 26.1 (CH₂), 26.5 (CH₂), 28.5

(CH₂), 31.6 (CH₂), 35.8 (CH₂), 37.8 (CH), 44.3 (CH), 48.0 (C), 50.4 (CH), 112.5 (CH_Ar), 125.8 (CH_Ar),

127.4 (C_Ar), 129.9 (CH_Ar), 130.23 (CH_Ar), 130.26 (CH_Ar), 130.3 (CH_Ar), 131.9 (C_Ar), 132.1 (C_Ar), 135.5

(C_Ar), 137.9 (C_Ar), 151.0 (C_Ar), 220.9 (C=O). IR (ATR, cm⁻¹):

(s), 1588 (w), 1473 (s), 1258 (m), 1165 (m), 1054 (m), 1008 (m), 816 (s). MS (EI, 70 eV): m/z (%) = 360

ν˜ = 3370 (w), 2923 (m), 2861 (w), 1734

15

(100) [M⁺], 361 (27), 275 (30), 262 (12), 236 (22), 195 (13). HRMS (EI, 70 eV): calcd. for C₂₅H₂₈O₂[M⁺]

360.20838; found 360.20841.

4-(p-tert-Butylphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (7 c): light yellow solid (104 mg, 90%),

mp = 150

–

151 °C, [α]_D= +71.5° (c 1.00, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.92 (s, 3H, CH₃),

1.32 – 1.38 (m, 1H, aliphatic), 1.38 (s, 9H, C(CH₃)₃), 1.49 – 1.61 (m, 5H, aliphatic), 1.87 – 2.16 (m, 4H,

aliphatic), 2.27 – 2.33 (m, 1H, aliphatic), 2.42 – 2.53 (m, 4H, aliphatic), 4.67 (br.s, 1H, OH), 6.86 (d, ³J_H-

= 8.5 Hz, 1H, CH_Ar), 7.18 – 7.26 (m, 3H, CH_Ar+ CDCl₃), 7.50 – 7.53 (m, 2H, CH_Ar). ¹³C NMR (75

H

MHz, CDCl₃): δ = 13.9 (CH₃), 21.5 (CH₂), 26.2 (CH₂), 26.5 (CH₂), 28.6 (CH₂), 31.4 (C(CH₃)₃), 31.6

(CH₂), 34.7 (C(CH₃)₃), 35.9 (CH₂), 37.9 (CH), 44.4 (CH), 48.0 (C), 50.4 (CH), 112.5 (CH_Ar), 125.7

(CH_Ar), 126.42 (CH_Ar), 126.47 (CH_Ar), 127.4 (C_Ar), 129.6 (CH_Ar), 130.1 (CH_Ar), 131.9 (C_Ar), 132.1 (C_Ar),

135.7 (C_Ar), 151.05 (C_Ar), 151.10 (C_Ar), 221.0 (C=O). IR (ATR, cm⁻¹):

˜ = 3306 (w), 2954 (m), 1718 (s),

ν

1585 (w), 1288 (s), 1201 (m), 1056 (m), 819 (s). MS (EI, 70 eV): m/z (%) = 402 (100) [M⁺], 403 (31),

387 (26), 317 (15), 278 (10), 57 (20), 55 (10), 41 (17). HRMS (EI, 70 eV): calcd. for C₂₈H₃₄O₂[M⁺]

402.25533; found 402.25473.

4-(p-Methoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (7 d): yellow solid (107 mg, 99%), mp =

1

97

–

98 °C, [α]_D= +60.0° (c 1.00, CHCl₃). H NMR (300 MHz, CDCl₃): δ = 0.92 (s, 3H, CH₃), 1.31 –

1.39 (m, 1H, aliphatic), 1.49 – 1.62 (m, 5H, aliphatic), 1.87 – 2.16 (m, 4H, aliphatic), 2.25 – 2.32 (m, 1H,

aliphatic), 2.42 – 2.53 (m, 4H, aliphatic), 3.87 (s, 3H, OCH₃), 4.69 (br.s, 1H, OH), 6.85 (d, ³J_H-H= 8.6 Hz,

1H, CH_Ar), 7.01 – 7.06 (m, 2H, CH_Ar), 7.16 – 7.26 (m, 3H, CH_Ar+ CDCl₃). ¹³C NMR (63 MHz, CDCl₃):

δ = 13.8 (CH₃), 21.5 (CH₂), 26.1 (CH₂), 26.6 (CH₂), 28.5 (CH₂), 31.6 (CH₂), 35.9 (CH₂), 37.8 (CH), 44.3

(CH), 48.0 (C), 50.4 (CH), 55.3 (OCH₃), 112.4 (CH_Ar), 114.9 (CH_Ar), 115.0 (CH_Ar), 125.8 (CH_Ar), 127.1

(2C_Ar), 131.2 (CH_Ar), 131.6 (CH_Ar), 131.9 (C_Ar), 135.8 (C_Ar), 151.2 (C_Ar), 159.4 (C_Ar), 221.0 (C=O). IR

(ATR, cm⁻¹):

˜ = 3391 (w), 2924 (m), 1733 (s), 1512 (s), 1472 (s), 1282 (m), 1242 (s), 1174 (s), 1031

ν

(s), 830 (s), 809 (s). MS (EI, 70 eV): m/z (%) = 376 (100) [M⁺], 377 (27), 291 (17), 252 (18). HRMS (EI,

70 eV): calcd. for C₂₅H₂₈O₃[M⁺] 376.20330; found 376.20304.

4-(p-Trifluoromethylphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (7 e): light brown solid (108 mg,

91%), mp = 104

–

105 °C, [α]_D= +85.3° (c 1.00, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.93 (s, 3H,

CH₃), 1.33 – 1.40 (m, 1H, aliphatic), 1.53 – 1.59 (m, 5H, aliphatic), 1.89 – 2.16 (m, 4H, aliphatic), 2.26 –

3

2.37 (m, 1H, aliphatic), 2.44 – 2.54 (m, 4H, aliphatic), 4.59 (br.s, 1H, OH), 6.84 (d, J_H-H= 8.5 Hz, 1H,

CH_Ar), 7.26 – 7.29 (m, 1H, CH_Ar+ CDCl₃), 7.39 – 7.44 (m, 2H, CH_Ar), 7.75 – 7.78 (m, 2H, CH_Ar). ¹³C

NMR (75 MHz, CDCl₃): δ = 13.8 (CH₃), 21.5 (CH₂), 26.1 (CH₂), 26.4 (CH₂), 28.6 (CH₂), 31.6 (CH₂),

1

35.8 (CH₂), 37.8 (CH), 44.3 (CH), 48.0 (C), 50.4 (CH), 113.1 (CH_Ar), 124.1 (q, J_C-F= 271.7 Hz, CF₃),

3

2

126.2 (m, J not given, 2CH_Ar), 126.40 (C_Ar), 126.44 (CH_Ar), 130.2 (q, J_C-F= 32.5 Hz, C_Ar-CF₃), 130.6

(CH_Ar), 130.9 (CH_Ar), 132.4 (C_Ar), 135.3 (C_Ar), 140.0 (C_Ar, 150.6 (C_Ar), 220.9 (C=O). ¹⁹F NMR

(282 MHz, CDCl₃): δ = 62.6 (CF₃). IR (ATR, cm⁻¹):

˜ = 3318 (w), 2926 (m), 1716 (m), 1616 (w) 1321

ν

16

(s), 1159 (s), 1117 (s), 1065 (s), 835 (m). MS (EI, 70 eV): m/z (%) = 414 (100) [M⁺], 415 (27), 357 (12),

330 (12), 329 (25), 316 (18), 304 (12), 303 (13), 301 (10), 291 (10), 290 (27), 289 (10), 55 (12), 41 (14).

HRMS (EI, 70 eV): calcd. for C₂₅H₂₅O₂F₃[M⁺] 414.18012; found 414.17903.

4-(p-Trifluoromethoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (7 f): light yellow solid (118

1

mg, 96%), mp = 94

–

95 °C, [α]_D= +83.3° (c 1.00, CHCl₃). H NMR (300 MHz, CDCl₃): δ = 0.92 (s,

3H, CH₃), 1.33 – 1.39 (m, 1H, aliphatic), 1.49 – 1.63 (m, 5H, aliphatic), 1.88 – 2.16 (m, 4H, aliphatic),

2.25 – 2.33 (m, 1H, aliphatic), 2.42 – 2.54 (m, 4H, aliphatic), 4.57 (br.s, 1H, OH), 6.85 (d, ³J_H-H= 8.6 Hz,

1H, CH_Ar), 7.25 – 7.38 (m, 5H, CH_Ar+ CDCl₃). ¹³C NMR (75 MHz, CDCl₃): δ = 13.8 (CH₃), 21.5

(CH₂), 26.1 (CH₂), 26.5 (CH₂), 28.9 (CH₂), 31.6 (CH₂), 35.8 (CH₂), 37.8 (CH), 44.3 (CH), 48.0 (C), 50.4

1

(CH), 112.9 (CH_Ar), 120.5 (q, J_C-F= 257.1 Hz, CF₃), 121.8 (2CH_Ar), 126.2 (C_Ar), 126.3 (CH_Ar), 131.7

3

(CH_Ar), 132.0 (CH_Ar), 132.3 (C_Ar), 134.4 (C_Ar), 135.5 (C_Ar), 149.0 (q, J_C-F= 1.9 Hz, C_Ar-OCF₃), 150.8

(C_Ar), 220.9 (C=O). ¹⁹F NMR (282 MHz, CDCl₃): δ = 57.7 (CF₃). IR (ATR, cm⁻¹

) ˜ = 3350 (w), 2928

ν

(w), 1723 (m), 1587 (w), 1475 (m), 1251 (s), 1154 (s), 1009 (m), 812 (m). MS (EI, 70 eV): m/z (%) = 430

(100) [M⁺], 431 (26), 346 (12), 345 (30), 332 (15), 320 (10), 319 (11), 306 (23), 69 (15), 55 (10), 41 (11).

HRMS (EI, 70 eV): calcd. for C₂₅H₂₅O₃F₃[M⁺] 430.17503; found 430.17490.

4-(m-Fluorophenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (7 g): light yellow solid (96 mg, 92%), mp

= 98

–

99 °C, [α]_D= +112.5° (c 1.00, CHCl₃). A mixture of diastereomers. ¹H NMR (300 MHz, CDCl₃):

δ = 0.92 and 0.93 (s, 3H, CH₃), 1.33 – 1.40 (m, 1H, aliphatic), 1.45 – 1.64 (m, 5H, aliphatic), 1.88 – 2.16

(m, 4H, aliphatic), 2.25 – 2.33 (m, 1H, aliphatic), 2.41 – 2.54 (m, 4H, aliphatic), 4.63 (br.s, 1H, OH), 6.85

(d, ³J_H-H= 8.6 Hz, 1H, CH_Ar), 6.97 – 7.16 (m, 3H, CH_Ar), 7.18 (d, ³J_H-H= 8.5 Hz, 1H, CH_Ar), 7.43 – 7.53

(m, 1H, CH_Ar). ¹³C NMR (75 MHz, CDCl₃): δ = 13.8 (CH₃), 21.5 (CH₂), 26.13 and 26.15 (CH₂), 26.5

(CH₂), 28.4 and 28.5 (CH₂), 31.6 (CH₂), 35.9 (CH₂), 37.78 and 37.80 (CH), 44.28 and 44.34 (CH), 48.0

2

(C), 50.4 (CH), 112.9 (CH_Ar), 115.0 and 115.3 (CH_Ar), 117.24 (d, J_C-F= 26.6 Hz) and 117.52 (d, J_C-F

=

2

26.6 Hz) (CH_Ar), 125.90 (d, J_C-F= 30.4 Hz) and 125.94 (d, J_C-F= 30.6 Hz) (CH_Ar), 126.25 and 126.29

3

(CH_Ar), 126.41 (C_Ar), 131.03 (d, J_C-F= 8.5 Hz) and 131.05 (d, J_C-F= 8.7 Hz) (CH_Ar), 132.17 (C_Ar),

3

135.30 and 135.33 (C_Ar), 137.95 (d, J_C-F= 7.4 Hz) and 137.97 (d, J_C-F= 7.5 Hz) (C_Ar), 150.7 (C_Ar),

163.33 (d, ¹J_C-F= 248.3 Hz) and 163.40 (d, ¹J_C-F= 248.1 Hz) (C_Ar-F), 220.9 (C=O). ¹⁹F NMR (282 MHz,

CDCl₃): δ = -111.45 and -111.53 (CF). IR (ATR, cm⁻¹):

˜ = 3340 (w), 2925 (m), 1723 (s), 1580 (s),

ν

1472 (s), 1261 (m), 1152 (m), 1055 (m), 785 (s). MS (EI, 70 eV): m/z (%) = 364 (100) [M⁺], 365 (27),

307 (10), 280 (10), 279 (27), 266 (16), 253 (11), 240 (26), 239 (11), 220 (14), 199 (11), 183 (12), 55 (10),

41 (14). HRMS (EI, 70 eV): calcd. for C₂₄H₂₅O₂F [M⁺] 364.18331; found 364.18327.

4-(3,5-Dimethoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (7 h): light yellow solid (102 mg,

1

88%), mp = 95

–

96 °C, [α]_D= +90.9° (c 1.00, CHCl₃). H NMR (300 MHz, CDCl₃): δ = 0.92 (s, 3H,

CH₃), 1.33 – 1.40 (m, 1H, aliphatic), 1.49 – 1.65 (m, 5H, aliphatic), 1.88 – 2.16 (m, 4H, aliphatic), 2.25 –

2.34 (m, 1H, aliphatic), 2.41 – 2.60 (m, 4H, aliphatic), 3.80 (s, 3H, OCH₃), 3.81 (s, 3H, OCH₃), 4.78 (br.s,

17

1H, OH), 6.40 – 6.43 (m, 2H, CH_Ar), 6.50 – 6.52 (m, 1H, CH_Ar), 6.85 (d, ³J_H-H= 8.6 Hz, 1H, CH_Ar), 7.23

– 7.26 (m, 1H, CH_Ar+ CDCl₃). ¹³C NMR (75 MHz, CDCl₃): δ = 13.8 (CH₃), 21.6 (CH₂), 26.1 (CH₂),

26.5 (CH₂), 28.2 (CH₂), 31.6 (CH₂), 35.9 (CH₂), 37.9 (CH), 44.3 (CH), 48.0 (C), 50.4 (CH), 55.4

(2OCH₃), 100.3 (CH_Ar), 107.5 (CH_Ar), 108.0 (CH_Ar), 112.6 (CH_Ar), 126.0 (CH_Ar), 127.5 (C_Ar), 131.9 (C_Ar),

135.2 (C_Ar), 137.3 (C_Ar), 150.7 (C_Ar), 161.7 (C_Ar), 161.8 (C_Ar), 220.9 (C=O). IR (ATR, cm⁻¹):

˜ = 3400

ν

(w), 2927 (w), 1733 (m), 1587 (s), 1452 (m), 1343 (m), 1202 (s), 1150 (s), 1060 (m), 817 (m). MS (EI, 70

eV): m/z (%) = 406 (100) [M⁺], 407 (27), 321 (17), 282 (12), 281 (10), 55 (11), 41 (12). HRMS (ESI, 70

eV): calcd. for C₂₆H₃₀O₄[M⁺] 406.21386; found 406.21368.

4-(p-Ethylphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (7 i): yellow solid (106 mg, 99%), mp =

110

–

111 °C, [α]_D= +78.5° (c 1.02, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.93 (s, 3H, CH₃), 1.31

(t, ³J_H-H= 7.6 Hz, 3H, CH₂CH₃), 1.28 – 1.35 (m, 1H, aliphatic), 1.49 – 1.61 (m, 5H, aliphatic), 1.87 – 2.16

3

(m, 4H, aliphatic), 2.26 – 2.34 (m, 1H, aliphatic), 2.42 – 2.53 (m, 4H, aliphatic), 2.73 (q, J_H-H= 7.6 Hz,

2H, CH₂CH₃), 4.59 (br.s, 1H, OH), 6.85 (d, ³J_H-H= 8.5 Hz, 1H, CH_Ar), 7.16 – 7.21 (m, 2H, CH_Ar), 7.23 –

7.26 (m, 1H, CH_Ar+ CDCl₃), 7.32 – 7.36 (m, 2H, CH_Ar). ¹³C NMR (75 MHz, CDCl₃): δ = 13.8 (CH₃),

15.4 (CH₂CH₃), 21.5 (CH₂), 26.1 (CH₂), 26.5 (CH₂), 28.5 (CH₂), 28.6 (CH₂CH₃), 31.6 (CH₂), 35.9 (CH₂),

37.8 (CH), 44.3 (CH), 48.0 (C), 50.4 (CH), 112.5 (CH_Ar), 125.7 (CH_Ar), 127.5 (C_Ar), 129.0 (CH_Ar), 129.03

(CH_Ar), 129.9 (CH_Ar), 130.4 (CH_Ar), 131.9 (C_Ar), 132.4 (C_Ar), 135.6 (C_Ar), 144.2 (C_Ar), 151.0 (C_Ar), 221.0

(C=O). IR (ATR, cm⁻¹):

˜ = 3369 (w), 2926 (m), 1734 (s), 1587 (m), 1472 (s), 1259 (s), 1165 (m), 830

ν

(s). MS (EI, 70 eV): m/z (%) = 374 (100) [M⁺], 375 (30), 289 (22), 276 (11), 250 (20). HRMS (EI, 70

eV): calcd. for C₂₆H₃₀O₂[M⁺] 374.22403; found 374.22323.

4-(p-Chlorophenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (7 j): light yellow solid (95 mg, 87%), mp

= 109

–

110 °C, [α]_D= +71.8° (c 1.01, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.92 (s, 3H, CH₃), 1.31

– 1.39 (m, 1H, aliphatic), 1.49 – 1.61 (m, 5H, aliphatic), 1.87 – 2.16 (m, 4H, aliphatic), 2.25 – 2.31 (m,

1H, aliphatic), 2.41 – 2.54 (m, 4H, aliphatic), 4.57 (br.s, 1H, OH), 6.84 (d, ³J_H-H= 8.6 Hz, 1H, CH_Ar), 7.18

– 7.27 (m, 3H, CH_Ar+ CDCl₃), 7.46 – 7.51 (m, 2H, CH_Ar). ¹³C NMR (75 MHz, CDCl₃): δ = 13.8 (CH₃),

21.5 (CH₂), 26.1 (CH₂), 26.5 (CH₂), 28.6 (CH₂), 31.6 (CH₂), 35.9 (CH₂), 37.8 (CH), 44.3 (CH), 48.0 (C),

50.4 (CH), 112.8 (CH_Ar), 126.2 (CH_Ar), 126.4 (C_Ar), 129.7 (2CH_Ar), 131.5 (CH_Ar), 131.9 (CH_Ar), 132.2

(C_Ar), 134.1 (C_Ar), 134.2 (C_Ar), 135.4 (C_Ar), 150.8 (C_Ar), 220.9 (C=O). IR (ATR, cm⁻¹):

˜ = 3344 (w),

ν

2926 (m), 1723 (s), 1586 (m), 1473 (s), 1285 (m), 1166 (m), 1088 (s), 1014 (m), 820 (s). MS (EI, 70 eV):

m/z (%) = 380 (100) [M⁺], 382 (37), 381 (26), 297 (11), 296 (10), 295 (28), 282 (18), 270 (10), 269 (12),

256 (26), 215 (13), 202 (13), 189 (11), 178 (10), 165 (11), 67 (10), 55 (16), 41 (17), 39 (10), 29 (10).

HRMS (EI, 70 eV): calcd. for C₂₄H₂₅O₂Cl [M⁺] 380.15376; found 380.15281; calcd. for C₂₄H₂₅O₂³⁷Cl

[M⁺] 382.15081; found 382.15141.

4-(o-Fluorophenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (7 k): brown solid (98 mg, 94%), mp = 95

–

1

96 °C, [α]_D= +82.1° (c 1.44, CHCl₃). A mixture of diastereomers. H NMR (300 MHz, CDCl₃): δ =

18

0.92 and 0.93 (2s, 3H, CH₃), 1.33 – 1.42 (m, 1H, aliphatic), 1.50 – 1.66 (m, 5H, aliphatic), 1.88 – 2.28 (m,

4H, aliphatic), 2.25 – 2.35 (m, 1H, aliphatic), 2.40 – 2.63 (m, 4H, aliphatic), 4.60 (br.s, 1H, OH), 6.85 (d,

³J_H-H= 8.6 Hz, 1H, CH_Ar), 7.19 – 7.29 (m, 4H, CH_Ar+ CDCl₃), 7.40 – 7.47 (m, 1H, CH_Ar). ¹³C NMR (75

MHz, CDCl₃): δ = 13.8 and 13.9 (CH₃), 21.6 (CH₂), 26.0 and 26.1 (CH₂), 26.5 (CH₂), 27.62 and 28.2

(CH₂), 31.64 and 31.66 (CH₂), 35.9 (CH₂), 37.73 and 37.83 (CH), 44.21 and 44.35 (CH), 48.00 and 48.03

(C), 50.44 and 50.48 (CH), 112.85 and 112.87 (CH_Ar), 116.48 (d, ²J_C-F= 22.2) and 116.57 (d, ²J_C-F= 22.3)

(CH_Ar), 121.24 and 121.31 (C_Ar), 122.95 (d, ²J_C-F= 17.8 Hz) and 122.98 (d, ²J_C-F= 17.6 Hz) (C_Ar), 124.95

3

– 125.02 (several signals, CH_Ar, C_Ar), 126.6 (CH_Ar), 130.39 (d, J_C-F= 8.0 Hz) and 130.41 (d, J_C-F= 8.1

Hz) (CH_Ar), 132.1 – 132.4 (several signals, CH_Ar), 136.21 and 136.37 (C_Ar), 151.06 and 151.16 (C_Ar),

160.1 (d, ¹J_C-F= 246.6 Hz) and 160.4 (2d, ¹J_C-F= 246.9 Hz) (C_Ar-F), 220.97 and 221.01 (C=O). ¹⁹F NMR

(75 MHz, CDCl₃): δ = -112.8 and -112.5 (CF). IR (ATR, cm⁻¹):

˜ = 3342 (w), 2925 (m), 1721 (s), 1588

ν

(w), 1451 (m), 1214 (m), 1056 (m), 755 (s). MS (EI, 70 eV): m/z (%) = 364 (100) [M⁺], 365 (29), 280

(11), 279 (30), 266 (16), 254 (10), 253 (14), 240 (32), 239 (14), 55 (10), 41 (11). HRMS (EI, 70 eV):

calcd. for C₂₄H₂₅O₂F [M⁺] 364.18331; found 364.18433.

4-(2,3,4-Trimethoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (7 l): light yellow solid (84 mg,

1

67%), mp = 121

–

122 °C, [α]_D= +82.5° (c 1.00, CHCl₃). A mixture of diastereomers. H NMR (300

MHz, CDCl₃): δ = 0.88 and 0.89 (2s, 3H, CH₃), 1.39 – 1.69 (m, 6H, aliphatic), 1.81 – 1.99 (m, 3H,

aliphatic), 2.05 – 2.13 (m, 1H, aliphatic), 2.24 – 2.29 (m, 1H, aliphatic), 2.34 – 2.52 (m, 4H, aliphatic),

3.58 and 3.60 (2s, 3H, OCH₃), 3.81 and 3.82 (2s, 3H, OCH₃), 3.87 (s, 3H, OCH₃), 6.68 – 6.76 (m, 2H,

CH_Ar), 6.83 – 6.88 (m, 1H, CH_Ar), 7.14 (d, ³J_H-H= 8.5 Hz, 1H, CH_Ar), 8.800 and 8.803 (2s, 1H, OH). ¹³

C

NMR (63 MHz, CDCl₃): δ = 13.8 and 13.9 (CH₃), 21.5 (CH₂), 25.9 and 26.1 (CH₂), 26.5 and 26.6 (CH₂),

27.5 and 28.4 (CH₂), 31.6 (CH₂), 35.9 (CH₂), 37.8 and 37.9 (CH), 44.3 (CH), 48.0 (C), 50.45 and 50.53

(CH), 56.0 (OCH₃), 61.1 (OCH₃), 61.2 (OCH₃), 108.1 and 108.2 (CH_Ar), 112.3 and 112.6 (CH_Ar), 121.25

and 121.33 (C_Ar), 123.8 and 123.9 (C_Ar), 125.6 and 125.9 (CH_Ar), 125.90 and 125.93 (CH_Ar), 131.9 and

132.1 (C_Ar), 136.3 and 136.6 (C_Ar), 142.9 and 143.1 (C_Ar), 151.1 and 151.2 (C_Ar), 152.0 (C_Ar), 154.0 (C_Ar),

C=O not given. IR (ATR, cm⁻¹):

˜ = 3415 (w), 2923 (m), 1725 (s), 1588 (w), 1461 (s), 1408 (m), 1271

ν

(s), 1093 (s), 1008 (s), 795 (m). MS (EI, 70 eV): m/z (%) = 436 (100) [M⁺], 437 (27). HRMS (EI, 70 eV):

calcd. for C₂₇H₃₂O₅[M⁺] 436.22443; found 436.22402.

4-(o-Ethoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (7 m): light yellow solid (88 mg, 79%), mp

1

= 144

–

145 °C, [α]_D= +77.7° (c 2.31, CHCl₃). A mixture of diastereomers. H NMR (300 MHz,

3

CDCl₃): δ = 0.92 and 0.93 (2s, 3H, CH₃), 1.26 and 1.27 (2t, J_H-H= 7.0 Hz, 3H, 2CH₂CH₃), 1.32 – 1.40

(m, 1H, aliphatic), 1.47 – 1.61 (m, 5H, aliphatic), 1.86 – 2.16 (m, 4H, aliphatic), 2.25 – 2.37 (m, 1H,

3

aliphatic), 2.42 – 2.53 (m, 4H, aliphatic), 4.05 (q, J_H-H= 7.0 Hz, 3H, 2CH₂CH₃), 4.67 and 4.72 (2br.s,

3

1H, OH), 6.84 and 6.85 (2d, J_H-H= 8.5 Hz, 1H, CH_Ar), 7.00 – 7.09 (m, 2H, CH_Ar), 7.14 – 7.18 (m, 1H,

CH_Ar), 7.22 – 7.26 (m, 1H, CH_Ar+ CDCl₃), 7.36 – 7.42 (m, 1H, CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ =

13.84 and 13.87 (CH₃), 14.6 and 14.7 (CH₃), 21.55 and 21.57 (CH₂), 25.9 and 26.1 (CH₂), 26.5 and 26.6

19

(CH₂), 27.2 and 28.3 (CH₂), 31.7 (CH₂), 35.9 (CH₂), 37.8 and 37.9 (CH), 44.2 and 44.4 (CH), 47.99 and

48.01 (C), 50.5 and 50.6 (CH), 63.8 and 64.0 (CH₂CH₃), 112.3 and 112.6 (CH_Ar), 112.99 and 113.07

(CH_Ar), 121.3 and 121.4 (CH_Ar), 124.1 and 124.2 (C_Ar), 124.2 and 124.4 (C_Ar), 125.5 and 125.7 (CH_Ar),

129.78 and 129.83 (CH_Ar), 131.6 and 131.8 (C_Ar), 132.0 and 132.3 (CH_Ar), 136.0 and 136.3 (C_Ar), 150.9

and 151.1 (C_Ar), 156.3 and 156.9 (C_Ar), 220.6 (C=O). IR (ATR, cm⁻¹):

˜ = 3397 (w), 2925 (w), 1733 (s),

ν

1590 (w), 1447 (s), 1341 (m), 1228 (s), 1040 (s), 922 (m), 753 (s). MS (EI, 70 eV): m/z (%) = 390 (100)

[M⁺], 391 (28), 305 (17), 266 (12), 29 (25). HRMS (ESI, 70 eV): calcd. for C₂₆H₃₀O₃[M⁺] 390.21895;

found 390.21851.

2-Phenyl-3-hydroxyestra-1,3,5(10)-trien-17-one (8 a):^11b2-Bromoestrone (6 b) (100 mg, 0.286 mmol),

boronic acid (1.5 equiv.), Pd(PPh₃)₄(17.0 mg, 0.05 equiv.), RuPhos (6.7 mg, 0.05 equiv.), K₂CO₃(79.1

mg, 2.0 equiv.) and dioxane (4 ml) were added in a pressure tube under argon atmosphere. The mixture

was stirred 16 h at 101°C. The solvent was evaporated in vacuo. The residue was purified by column

chromatography (silica gel, heptane/EtOAc = 1:5). 8a was isolated as a white solid (74 mg, 75%), mp =

197

–

198 °C (lit. 169 °C), [α]_D= +161.1° (c 1.00, CHCl₃) (lit. [α]_D= +112.0° (c 1.00, CHCl₃). ¹H NMR

(250 MHz, CDCl₃): δ = 0.93 (s, 3H, CH₃), 1.42 – 1.70 (m, 6H, aliphatic), 1.93 – 2.20 (m, 4H, aliphatic),

2.25 – 2.35 (m, 1H, aliphatic), 2.37 – 2.57 (m, 2H, aliphatic), 2.91 – 2.96 (m, 2H, aliphatic), 5.17 (br.s,

1H, OH), 6.75 (s, 1H, CH_Ar), 7.17 (s, 1H, CH_Ar), 7.34 – 7.41 (m, 1H, CH_Ar), 7.46 – 7.48 (m, 4H, CH_Ar).

¹³C NMR (63 MHz, CDCl₃): δ = 13.9 (CH₃), 21.6 (CH₂), 26.0 (CH₂), 26.5 (CH₂), 29.3 (CH₂), 31.6

(CH₂), 35.9 (CH₂), 38.4 (CH), 44.0 (CH), 48.0 (C), 50.4 (CH), 115.7 (CH_Ar), 125.8 (C_Ar), 127.2 (CH_Ar),

127.6 (CH_Ar), 129.1 (2CH_Ar), 129.2 (2CH_Ar), 132.2 (C_Ar), 137.4 (C_Ar), 137.7 (C_Ar), 150.4 (C_Ar), 221.1

(C=O). IR (ATR, cm⁻¹):

˜ = 3329 (w), 2928 (m), 1723 (s), 1614 (w), 1409 (s), 1200 (m), 893 (m), 769

ν

(s), 698 (s). MS (EI, 70 eV): m/z (%) = 346 (100) [M⁺], 347 (23), 261 (25), 248 (13), 222 (23), 221 (11),

202 (13), 178 (11), 165 (12), 55 (10), 41 (12). HRMS (ESI, 70 eV): calcd. for C₂₄H₂₆O₂[M⁺] 346.19273;

found 346.19267.

2-(p-Tolyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (8 b):^11bwhite solid (77 mg, 75%), mp = 201

– 202

°C, [α]_D= +160.5° (c 1.00, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.93 (s, 3H, CH₃), 1.47 – 1.65 (m,

6H, aliphatic), 1.93 – 2.19 (m, 4H, aliphatic), 2.25 – 2.33 (m, 1H, aliphatic), 2.38 – 2.56 (m, 2H,

aliphatic), 2.41 (s, 3H, CH₃), 2.91 – 2.95 (m, 2H, aliphatic), 5.26 (br.s, 1H, OH), 6.74 (s, 1H, CH_Ar), 7.16

3

(s, 1H, CH_Ar), 7.26 – 7.30 (m, 2H, CH_Ar+ CDCl₃), 7.36 (d, J_H-H= 7.9 Hz, 2H, CH_Ar). ¹³C NMR (63

MHz, CDCl₃): δ = 13.9 (CH₃), 21.2 (CH₃), 21.6 (CH₂), 26.0 (CH₂), 26.6 (CH₂), 29.3 (CH₂), 31.6 (CH₂),

35.9 (CH₂), 38.4 (CH), 44.0 (C), 48.0 (CH), 50.4 (CH), 115.6 (CH_Ar), 125.8 (C_Ar), 127.2 (CH_Ar), 128.9

(2CH_Ar), 129.9 (2CH_Ar), 132.1 (C_Ar), 134.4 (C_Ar), 137.4 (C_Ar), 137.5 (C_Ar), 150.5 (C_Ar), 221.1 (C=O). IR

(ATR, cm⁻¹):

˜ = 3342 (w), 2925 (m), 1723 (s), 1491 (s), 1237 (s), 1051 (m), 822 (s). MS (EI, 70 eV):

ν

m/z (%) = 360 (100) [M⁺], 361 (26), 275 (19), 236 (18). HRMS (ESI, 70 eV): calcd. for C₂₅H₂₈O₂[M⁺]

360.20838; found 360.20759.

20

2-(p-tert-Butylphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (8 c): white solid (94 mg, 82%), mp =

229

–

230 °C, [α]_D= +129.6° (c 1.00, CHCl₃). ¹H NMR (250 MHz, CDCl₃): δ = 0.92 (s, 3H, CH₃), 1.37

(s, 9H, C(CH₃)₃), 1.47 – 1.66 (m, 6H, aliphatic), 1.92 – 2.19 (m, 4H, aliphatic), 2.24 – 2.34 (m, 1H,

aliphatic), 2.34 – 2.57 (m, 2H, aliphatic), 2.90 – 2.95 (m, 2H, aliphatic), 5.17 (br.s, 1H, OH), 6.74 (s, 1H,

CH_Ar), 7.17 (s, 1H, CH_Ar), 7.37 – 7.41 (m, 2H, CH_Ar), 7.48 – 7.52 (m, 2H, CH_Ar). ¹³C NMR (63 MHz,

CDCl₃): δ = 13.9 (CH₃), 21.6 (CH₂), 26.0 (CH₂), 26.6 (CH₂), 29.3 (CH₂), 31.3 (C(CH₃)₃), 31.6 (CH₂),

34.6 (C(CH₃)₃), 35.9 (CH₂), 38.4 (CH), 44.0 (CH), 48.0 (C), 50.4 (CH), 115.6 (CH_Ar), 125.7 (C_Ar), 126.2

(2CH_Ar), 127.2 (CH_Ar), 128.7 (2CH_Ar), 132.1 (C_Ar), 134.3 (C_Ar), 137.5 (C_Ar), 150.5 (C_Ar), 150.7 (C_Ar),

221.0 (C=O). IR (ATR, cm⁻¹):

˜ = 3347 (w), 2920 (m), 1723 (s), 1613 (w), 1500 (m), 1395 (m), 1257

ν

(m), 1193 (s), 896 (m), 828 (s), 752 (s). MS (EI, 70 eV): m/z (%) = 402 (100) [M⁺], 403 (32), 388 (26),

287 (86), 57 (14), 41 (14). HRMS (ESI, 70 eV): calcd. for C₂₈H₃₄O₂[M⁺] 402.25533; found 402.25513.

2-(p-Methoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (8 d): light yellow solid (86 mg, 80%),

mp = 183

–

184 °C, [α]_D= +137.0° (c 1.00, CHCl₃). ¹H NMR (250 MHz, CDCl₃): δ = 0.92 (s, 3H, CH₃),

1.47 – 1.64 (m, 6H, aliphatic), 1.92 – 2.19 (m, 4H, aliphatic), 2.22 – 2.32 (m, 1H, aliphatic), 2.37 – 2.57

(m, 2H, aliphatic), 2.89 – 2.94 (m, 2H, aliphatic), 3.85 (s, 3H, OCH₃), 5.15 (br.s, 1H, OH), 6.73 (s, 1H,

CH_Ar), 6.97 – 7.03 (m, 2H, CH_Ar), 7.14 (s, 1H, CH_Ar), 7.35 – 7.41 (m, 2H, CH_Ar). ¹³C NMR (63 MHz,

CDCl₃): δ = 13.9 (CH₃), 21.6 (CH₂), 26.0 (CH₂), 26.5 (CH₂), 29.2 (CH₂), 31.6 (CH₂), 35.9 (CH₂), 38.4

(CH), 44.0 (CH), 48.0 (C), 50.4 (CH), 55.4 (OCH₃), 114.6 (2CH_Ar), 115.6 (CH_Ar), 125.5 (C_Ar), 127.2

(CH_Ar), 129.5 (C_Ar), 130.2 (2CH_Ar), 132.1 (C_Ar), 137.3 (C_Ar), 150.5 (C_Ar), 159.2 (C_Ar), 221.0 (C=O). IR

(ATR, cm⁻¹):

˜ = 3350 (w), 2925 (m), 1720 (s), 1609 (m), 1501 (s), 1242 (s), 1177 (s), 1033 (s), 828 (s).

ν

MS (EI, 70 eV): m/z (%) = 376 (100) [M⁺], 377 (28), 291 (11), 252 (11). HRMS (ESI, 70 eV): calcd. for

C₂₅H₂₈O₃[M⁺] 376.20330; found 376.20276.

2-(p-Trifluoromethylphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (8 e): white solid (85 mg, 72%),

mp = 250

–

251 °C, [α]_D= +123.6° (c 1.00, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.93 (s, 3H, CH₃),

1.48 – 1.65 (m, 6H, aliphatic), 1.94 – 2.19 (m, 4H, aliphatic), 2.27 – 2.34 (m, 1H, aliphatic), 2.39 – 2.57

(m, 2H, aliphatic), 2.91 – 2.94 (m, 2H, aliphatic), 5.18 (br.s, 1H, OH), 6.73 (s, 1H, CH_Ar), 7.18 (s, 1H,

3

CH_Ar), 7.62 (d, J_H-H= 8.1 Hz, 2H, CH_Ar), 7.70 (d, J_H-H= 8.1 Hz, 2H, CH_Ar). ¹³C NMR (75 MHz,

CDCl₃): δ = 13.9 (CH₃), 21.6 (CH₂), 26.0 (CH₂), 26.5 (CH₂), 29.2 (CH₂), 31.5 (CH₂), 35.9 (CH₂), 38.3

(CH), 43.9 (CH), 48.0 (C), 50.4 (CH), 116.2 (CH_Ar), 124.6 (C_Ar), 124.2 (q, ¹J_C-F= 272.0 Hz, CF₃), 125.7

3

2

(q, J_C-F= 3.7 Hz, 2CH_Ar), 127.4 (CH_Ar), 129.41 (q, J_C-F= 32.6 Hz, C_Ar-CF₃), 129.48 (2CH_Ar), 132.7

(C_Ar), 138.5 (C_Ar), 141.6 (C_Ar), 150.4 (C_Ar), 221.2 (C=O). ¹⁹F NMR (282 MHz, CDCl₃): δ = -62.5 (CF₃).

IR (ATR, cm⁻¹):

˜ = 3365 (w), 2937 (w), 1722 (s), 1612 (m), 1324 (s), 1109 (s), 1067 (s), 1013 (m), 840

ν

(s). MS (EI, 70 eV): m/z (%) = 414 (100) [M⁺], 415 (28), 357 (12), 330 (10), 329 (23), 316 (17), 304 (11),

303 (14), 301 (12), 290 (22), 289 (12), 55 (12), 41 (13). HRMS (ESI, 70 eV): calcd. for C₂₅H₂₅O₂F₃[M⁺]

414.18012; found 414.17980.

21

2-(p-Trifluoromethoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (8 f): white solid (95 mg, 77%),

mp = 109

–

110 °C, [α]_D= +122.2° (c 1.00, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.92 (s, 3H, CH₃),

1.47 – 1.64 (m, 6H, aliphatic), 1.93 – 2.19 (m, 4H, aliphatic), 2.25 – 2.33 (m, 1H, aliphatic), 2.38 – 2.56

(m, 2H, aliphatic), 2.90 – 2.94 (m, 2H, aliphatic), 5.06 (br.s, 1H, OH), 6.72 (s, 1H, CH_Ar), 7.15 (s, 1H,

3

CH_Ar), 7.30 (d, J_H-H= 8.1 Hz, 2H, CH_Ar), 7.48 – 7.53 (m, 2H, CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ =

13.8 (CH₃), 21.6 (CH₂), 26.0 (CH₂), 26.5 (CH₂), 29.2 (CH₂), 31.5 (CH₂), 35.9 (CH₂), 38.3 (CH), 43.9

1

(CH), 48.0 (C), 50.4 (CH), 116.0 (CH_Ar), 120.5 (q, J_C-F= 257.5 Hz, CF₃), 121.4 (2CH_Ar), 124.6 (C_Ar),

3

127.4 (CH_Ar), 130.6 (2CH_Ar), 132.5 (C_Ar), 136.4 (C_Ar), 138.1 (C_Ar), 148.5 (q, J_C-F= 2.3 Hz, C_Ar-OCF₃),

150.3 (C_Ar), 220.9 (C=O). ¹⁹F NMR (282 MHz, CDCl₃): δ = -57.8 (CF₃). IR (ATR, cm⁻¹):

˜ = 3334 (w),

ν

2924 (m), 1721 (s), 1615 (w), 1499 (m), 1249 (s), 1202 (s), 1155 (s). MS (EI, 70 eV): m/z (%) = 430

(100) [M⁺], 431 (24), 346 (10), 345 (21), 332 (14), 319 (10), 306 (19), 69 (19), 55 (10). HRMS (ESI, 70

eV): calcd. for C₂₅H₂₅O₃F₃[M⁺] 430.17503; found 430.17478.

2-(m-Fluorophenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (8 g): light yellow solid (82 mg, 79%), mp

1

= 132

–

133 °C, [α]_D= +129.8° (c 1.00, CHCl₃). H NMR (300 MHz, CDCl₃): δ = 0.93 (s, 3H, CH₃),

1.48 – 1.64 (m, 6H, aliphatic), 1.94 – 2.19 (m, 4H, aliphatic), 2.25 – 2.39 (m, 1H, aliphatic), 2.39 – 2.56

(m, 2H, aliphatic), 2.90 – 2.94 (m, 2H, aliphatic), 5.20 (br.s, 1H, OH), 6.73 (s, 1H, CH_Ar), 7.03 – 7.09 (m,

1H, CH_Ar), 7.16 – 7.27 (m, 3H, CH_Ar+ CDCl₃), 7.38 – 7.46 (m, 1H, CH_Ar). ¹³C NMR (75 MHz, CDCl₃):

δ = 13.9 (CH₃), 21.6 (CH₂), 26.0 (CH₂), 26.5 (CH₂), 29.2 (CH₂), 31.5 (CH₂), 35.9 (CH₂), 38.3 (CH), 43.9

2

(CH), 48.0 (C), 50.4 (CH), 114.4 (d, J_C-F= 21.0 Hz, CH_Ar), 116.0 (CH_Ar), 116.2 (d, J_C-F= 21.6 Hz,

4

3

CH_Ar), 124.65 (d, J_C-F= 3.0 Hz, CH_Ar), 124.7 (d, J_C-F= 2.0 Hz, C_Ar), 127.2 (CH_Ar), 130.5 (d, J_C-F= 8.5

Hz, CH_Ar), 132.4 (C_Ar), 138.2 (C_Ar), 139.9 (d, ³J_C-F= 7.8 Hz, C_Ar), 150.3 (C_Ar), 163.1 (d, ¹J_C-F= 247.0 Hz,

C_Ar-F), 221.2 (C=O). ¹⁹F NMR (282 MHz, CDCl₃): δ = -112.1 (CF). IR (ATR, cm⁻¹):

˜ = 3348 (w),

ν

2921 (m), 1721 (s), 1612 (m), 1579 (m), 1402 (s), 1254 (s), 1012 (m), 874 (s), 782 (s). MS (EI, 70 eV):

m/z (%) = 364 (100) [M⁺], 365 (25), 279 (22), 266 (17), 253 (12), 251 (12), 240 (19), 239 (11), 220 (10),

55 (11), 41 (14). HRMS (ESI, 70 eV): calcd. for C₂₄H₂₅O₂F [M⁺] 364.18331; found 364.18303.

2-(3,5-Dimethoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (8 h): light yellow solid (116 mg,

1

84%), mp = 115

–

116 °C, [α]_D= +161.6° (c 1.00, CHCl₃). H NMR (300 MHz, CDCl₃): δ = 0.92 (s,

3H, CH₃), 1.47 – 1.64 (m, 6H, aliphatic), 1.93 – 2.18 (m, 4H, aliphatic), 2.25 – 2.33 (m, 1H, aliphatic),

2.37 – 2.56 (m, 2H, aliphatic), 2.90 – 2.94 (m, 2H, aliphatic), 3.82 (s, 6H, 2OCH₃), 5.32 (br.s, 1H, OH),

4

6.48 (pt, J_H-H= 2.3 Hz, 1H, CH_Ar), 6.57 (d, J_H-H= 2.3 Hz, 2H, CH_Ar), 6.74 (s, 1H, CH_Ar), 7.17 (s, 1H,

CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ = 13.8 (CH₃), 21.6 (CH₂), 26.0 (CH₂), 26.5 (CH₂), 29.3 (CH₂),

31.5 (CH₂), 35.9 (CH₂), 38.4 (CH), 43.9 (CH), 48.0 (C), 50.4 (CH), 55.4 (2OCH₃), 99.7 (CH_Ar), 106.9

(2CH_Ar), 115.6 (CH_Ar), 125.6 (C_Ar), 126.8 (CH_Ar), 132.1 (C_Ar), 137.9 (C_Ar), 139.3 (C_Ar), 150.3 (C_Ar), 161.5

(2C_Ar), 221.0 (C=O). IR (ATR, cm⁻¹):

˜ = 3369 (w), 2924 (m), 1720 (s), 1591 (s), 1453 (m), 1399 (s),

ν

1341 (m), 1192 (s), 1149 (s), 1057 (s), 833 (s). MS (EI, 70 eV): m/z (%) = 406 (100) [M⁺], 407 (28), 321

(11), 282 (10). HRMS (ESI, 70 eV): calcd. for C₂₆H₃₀O₄[M⁺] 406.21386; found 406.21387.

22

2,4-Diphenyl-3-hydroxyestra-1,3,5(10)-trien-17-one (9 a): 2,4-Dibromoestrone (6 c) (100 mg, 0.234

mmol), boronic acid (3 equiv.), Pd(PPh₃)₄(13.5 mg, 0.05 equiv.), RuPhos (5.5 mg, 0.05 equiv.), K₂CO₃

(129.4 mg, 4.0 equiv.) and dioxane (4 ml) were added in a pressure tube under argon atmosphere. The

mixture was stirred 40 h (or 72 h, see Table 4) at 101 °C. The solvent was evaporated in vacuo. The

residue was purified by column chromatography (silica gel, heptane/EtOAc = 1:4). 9a was isolated as a

1

light yellow solid (98 mg, 99%), mp = 110

–

111 °C (lit.⁸118 °C), [α]_D= +81.3° (c 1.00, CHCl₃). H

NMR (300 MHz, CDCl₃): δ = 0.95 (s, 3H, CH₃), 1.38 – 1.67 (m, 6H, aliphatic), 1.91 – 2.20 (m, 4H,

aliphatic), 2.33 – 2.41 (m, 1H, aliphatic), 2.46 – 2.57 (m, 4H, aliphatic), 4.84 (s, 1H, OH), 7.31 – 7.37 (m,

4H, CH_Ar), 7.42 – 7.47 (m, 3H, CH_Ar), 7.50 – 7.60 (m, 4H, CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ = 13.9

(CH₃), 21.6 (CH₂), 26.2 (CH₂), 26.6 (CH₂), 28.6 (CH₂), 31.6 (CH₂), 35.9 (CH₂), 37.9 (CH), 44.3 (CH),

48.0 (C), 50.4 (CH), 125.8 (C_Ar), 127.0 (CH_Ar), 127.2 (CH_Ar), 128.1 (CH_Ar), 128.2 (C_Ar), 128.5 (2CH_Ar),

129.3 (2CH_Ar), 129.4 (2CH_Ar), 130.1 (CH_Ar), 130.4 (CH_Ar), 132.1 (C_Ar), 135.1 (C_Ar), 135.9 (C_Ar), 138.2

(C_Ar), 147.7 (C_Ar), 220.9 (C=O). IR (ATR, cm⁻¹):

˜ = 3530 (w), 2922 (m), 1734 (s), 1601 (w), 1461 (m),

ν

1438 (m), 1410 (m), 1230 (m), 1145 (m), 755 (m), 699 (s) . MS (EI, 70 eV): m/z (%) = 422 (100) [M⁺],

423 (40), 337 (14), 298 (13), 265 (13), 41 (12). HRMS (ESI, 70 eV): calcd. for C₃₀H₃₀O₂[M⁺] 422.22403;

found 422.22350.

2,4-Di-(p-tolyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 b): light yellow solid (96 mg, 91%), mp =

134

–

135 °C, [α]_D= +72.2° (c 1.02, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.94 (s, 3H, CH₃), 1.46 –

1.64 (m, 5H, aliphatic), 1.91 – 2.17 (m, 4H, aliphatic), 2.40 (s, 3H, CH₃), 2.43 (s, 3H, CH₃), 2.27 – 2.57

3

(m, 6H, aliphatic), 4.87 (s, 1H, OH), 7.19 – 7.34 (m, 7H, CH_Ar+ CDCl₃), 7.47 (d, J_H-H= 8.1 Hz, 2H,

CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ = 13.8 (CH₃), 21.2 (CH₃), 21.3 (CH₃), 21.6 (CH₂), 26.2 (CH₂),

26.6 (CH₂), 28.6 (CH₂), 31.6 (CH₂), 35.9 (CH₂), 37.9 (CH), 44.4 (CH), 48.0 (C), 50.4 (CH), 125.6 (C_Ar),

126.8 (CH), 128.0 (C), 129.1 (2CH), 129.2 (2CH), 129.9 (CH), 130.09 (CH), 130.10 (CH), 130.2 (CH),

131.9 (C_Ar), 132.7 (C_Ar), 135.0 (C_Ar), 135.3 (C_Ar), 136.8 (C_Ar), 137.7 (C_Ar), 147.9 (C_Ar), 221.0 (C=O). IR

(ATR, cm⁻¹):

˜ = 3526 (w), 2921 (m), 2860 (m), 1737 (s), 1512 (m), 1454 (s), 1256 (m), 1232 (m), 1145

ν

(m), 818 (s). MS (EI, 70 eV): m/z (%) = 450 (100) [M⁺], 451 (38), 365 (9), 326 (9), 285 (8). HRMS (ESI,

70 eV): calcd. for C₃₂H₃₄O₂[M⁺] 450.25533; found 450.25665.

2,4-Di-(p-tert-butylphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 c): yellow solid (106 mg, 85%),

mp = 141

–

142 °C, [α]_D= +66.1° (c 1.00, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.94 (s, 3H, CH₃),

1.36 (s, 9H, C(CH₃)₃), 1.40 (s, 9H, C(CH₃)₃), 1.50 – 1.64 (m, 6H, aliphatic), 1.95 – 2.16 (m, 4H,

aliphatic), 2.33 – 2.39 (m, 1H, aliphatic), 2.45 – 2.54 (m, 4H, aliphatic), 4.91 (s, 1H, OH), 7.23 – 7.25 (m,

2H, CH_Ar+ CDCl₃), 7.31 (s, 1H, CH_Ar), 7.43 – 7.53 (m, 6H, CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ =

13.9 (CH₃), 21.6 (CH₂), 26.2 (CH₂), 26.6 (CH₂), 28.6 (CH₂), 31.4 (2C(CH₃)₃), 31.6 (CH₂), 34.5

(C(CH₃)₃), 34.7 (C(CH₃)₃), 35.9 (CH₂), 37.9 (CH), 44.4 (CH), 48.0 (C), 50,4 (CH), 114.7 (CH_Ar), 125.4

(CH_Ar), 125.5 (C_Ar), 126.25 (CH_Ar), 126.30 (CH_Ar), 126.35 (CH_Ar), 126.8 (CH_Ar), 128.0 (C_Ar), 128.9

23

(CH_Ar), 129.6 (CH_Ar), 130.0 (CH_Ar), 131.9 (C_Ar), 132.6 (C_Ar), 135.0 (C_Ar), 135.3 (C_Ar), 148.0 (C_Ar), 149.9

(C_Ar), 150.9 (C_Ar), 221.0 (C=O). IR (ATR, cm⁻¹):

˜ = 3530 (w), 2953 (m), 1738 (s), 1610 (w), 1515 (w),

ν

1456 (s), 1259 (s), 1149 (m), 1017 (m), 830 (s). MS (EI, 70 eV): m/z (%) = 534 (100) [M⁺], 535 (43), 520

(15), 519 (38), 57 (31), 41 (11). HRMS (ESI, 70 eV): calcd. for C₃₈H₄₆O₂[M⁺] 534.34923; found

534.34874.

2,4-Di-(p-methoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 d): light yellow solid (112 mg,

99%), mp = 117

–

118 °C, [α]_D= +84.2° (c 1.00, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.94 (s, 3H,

CH₃), 1.46 – 1.64 (m, 6H, aliphatic), 1.91 – 2.17 (m, 4H, aliphatic), 2.32 – 2.39 (m, 1H, aliphatic), 2.46 –

2.54 (m, 4H, aliphatic), 3.84 (s, 3H, OCH₃), 3.87 (s, 3H, OCH₃), 4.87 (s, 1H, OH), 6.97 (d, ³J_H-H= 8.7 Hz,

3

2H, CH_Ar), 7.05 (d, J_H-H= 8.2 Hz, 2H, CH_Ar), 7.21 – 7.28 (m, 3H, CH_Ar), 7.51 (d, J_H-H= 8.7 Hz, 2H,

CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ = 13.8 (CH₃), 21.5 (CH₂), 26.2 (CH₂), 26.6 (CH₂), 28.6 (CH₂),

31.6 (CH₂), 35.9 (CH₂), 37.9 (CH), 44.4 (CH), 48.0 (C), 50.4 (CH), 55.29 (OCH₃), 55.30 (OCH₃), 113.9

(2CH_Ar), 114.8 (CH_Ar), 114.9 (CH_Ar), 125.3 (C_Ar), 126.7 (CH_Ar), 127.6 (C_Ar), 127.7 (C_Ar), 130.3 (2CH_Ar),

130.6 (C_Ar), 131.2 (CH_Ar), 131.6 (CH_Ar), 131.9 (C_Ar), 135.0 (C_Ar), 148.1 (C_Ar), 158.8 (C_Ar), 159.3 (C_Ar),

220.9 (C=O). IR (ATR, cm⁻¹):

˜ = 3521 (w), 2925 (m), 1734 (s), 1607 (m), 1510 (s), 1456 (s), 1241 (s),

ν

1174 (s), 1031 (s), 829 (s). MS (EI, 70 eV): m/z (%) = 482 (100) [M⁺], 483 (33), 397 (3), 358 (5), 317 (5).

HRMS (ESI, 70 eV): calcd. for C₃₂H₃₄O₄[M⁺] 482.24516; found 482.24510.

2,4-Di-(p-trifluoromethylphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 e): yellow solid (116 mg,

89%), mp = 127

–

128 °C, [α]_D= +52.1° (c 1.55, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.94 (s, 3H,

CH₃), 1.37 – 1.64 (m, 6H, aliphatic), 1.97 – 2.17 (m, 4H, aliphatic), 2.36 – 2.48 (m, 5H, aliphatic), 4.73

(s, 1H, OH), 7.33 (s, 1H, CH_Ar), 7.45 – 7.49 (m, 2H, CH_Ar), 7.69 (m, 4H, CH_Ar), 7.79 – 7.81 (m, 2H,

CH_Ar). ¹³C NMR (75 MHz, CDCl₃): δ = 13.8 (CH₃), 21.5 (CH₂), 26.2 (CH₂), 26.4 (CH₂), 28.6 (CH₂),

1

31.6 (CH₂), 35.8 (CH₂), 37.8 (CH), 44.2 (CH), 47.9 (C), 50.4 (CH), 124.3 (q, J_C-F= 272.2 Hz, CF₃),

3

1

124.8 (C_Ar), 125.4 (q, J_C-F= 3.7 Hz, 2CH_Ar), 125.6 (q, J_C-F= 272.4 Hz, CF₃), 126.36 (CH_Ar), 126.39

2

(CH_Ar), 127.2 (C_Ar), 127.5 (CH_Ar), 129.3 (q, J_C-F= 32.4 Hz, C_Ar), 129.6 (2CH_Ar), 130.5 (q, J_C-F= 32.6

Hz, C_Ar), 130.7 (CH_Ar), 130.9 (CH_Ar), 132.9 (C_Ar), 135.8 (C_Ar), 139.8 (C_Ar), 141.7 (C_Ar), 147.5 (C_Ar),

220.6 (C=O). ¹⁹F NMR (282 MHz, CDCl₃): δ = -62.6 (CF₃), -62.5 (CF₃). IR (ATR, cm⁻¹):

˜ = 3275 (w),

ν

2928 (w), 1725 (m), 1615 (m), 1461 (m), 1398 (m), 1319 (s), 1160 (s), 1105 (s), 1064 (s), 1016 (s), 838

(m). MS (EI, 70 eV): m/z (%) = 558 (100) [M⁺], 559 (34), 539 (11), 473 (15), 460 (12), 434 (17), 12 (55).

HRMS (ESI, 70 eV): calcd. for C₃₂H₂₈O₂F₆[M⁺] 558.19880; found 558.19935.

2,4-Di-(p-trifluormethoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 f): light yellow solid

(109 mg, 79%), mp = 97

–

98 °C, [α]_D= +73.1° (c 1.00, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.94

(s, 3H, CH₃), 1.39 – 1.64 (m, 6H, aliphatic), 1.97 – 2.17 (m, 4H, aliphatic), 2.32 – 2.39 (m, 1H, aliphatic),

2.46 – 2.55 (m, 4H, aliphatic), 4.69 (s, 1H, OH), 7.26 – 7.29 (m, 3H, CH_Ar+ CDCl₃), 7.36 – 7.37 (m, 4H,

CH_Ar), 7.58 (d, ³J_H-H= 8.6 Hz, 2H, CH_Ar). ¹³C NMR (75 MHz, CDCl₃): δ = 13.8 (CH₃), 21.5 (CH₂), 26.2

24

(CH₂), 26.5 (CH₂), 28.6 (CH₂), 31.6 (CH₂), 35.8 (CH₂), 37.8 (CH), 44.3 (CH), 47.9 (C), 50.4 (CH),

1

120.49 (q, J_C-F= 257.7 Hz, OCF₃), 120.52 (q, J_C-F= 257.0 Hz, OCF₃), 120.9 (2CH_Ar), 121.8 (2CH_Ar),

124.6 (C_Ar), 127.0 (C_Ar), 127.3 (CH_Ar), 130.7 (2CH_Ar), 131.7 (CH_Ar), 132.0 (CH_Ar), 132.6 (C_Ar), 134.3

(C_Ar), 135.6 (C_Ar), 136.7 (C_Ar), 147.6 (C_Ar), 148.4 (C_Ar), 149.1 (C_Ar), 220.7 (C=O). ¹⁹F NMR (282 MHz,

CDCl₃): δ = 57.8 (CF₃), -57.7 (CF₃). IR (ATR, cm⁻¹):

˜ = 3545 (w), 2928 (w), 1732 (m), 1507 (m), 1460

ν

(m), 1250 (s), 1203 (s), 1153 (s), 1019 (m), 831 (m). MS (EI, 70 eV): m/z (%) = 590 (100) [M⁺], 591

(33), 505 (19), 466 (14), 69 (18), 55 (12). HRMS (ESI, 70 eV): calcd. for C₃₂H₂₈O₄F₆[M⁺] 590.18863;

found 590.18736.

2,4-Di-(m-fluorophenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 g): light yellow solid (91 mg, 85%),

1

mp = 111

–

112 °C, [α]_D= +108.0° (c 1.00, CHCl₃). A mixture of diastereomers. H NMR (300 MHz,

CDCl₃): δ = 0.939 and 0.943 (2s, 3H, CH₃), 1.37 – 1.65 (m, 6H, aliphatic), 1.92 – 2.20 (m, 4H, aliphatic),

2.31 – 2.39 (m, 1H, aliphatic), 2.46 – 2.55 (m, 4H, aliphatic), 4.80 and 4.81 (s, 1H, OH), 7.00 – 7.17 (m,

4H, CH_Ar), 7.27 – 7.43 (m, 4H, CH_Ar), 7.46 – 7.54 (m, 1H, CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ = 13.8

(CH₃), 21.5 (CH₂), 26.2 (CH₂), 26.5 (CH₂), 28.4 and 28.5 (CH₂), 31.6 (CH₂), 35.8 (CH₂), 37.8 (CH), 44.2

2

and 44.3 (CH), 47.9 (C), 50.4 (CH), 114.1 (d, J_C-F= 21.1 Hz, CH_Ar), 115.2 (d, J_C-F= 21.0 Hz, CH_Ar),

116.1 and 116.5 (CH_Ar), 117.2 (d, ²J_C-F= 20.9 Hz) and 117.5 (d, ²J_C-F= 21.0 Hz) (CH_Ar), 124.66 – 124.72

4

2

(several signals, C_Ar), 124.8 (d, J_C-F= 2.8 Hz, CH_Ar), 125.85 (d, J_C-F= 20.5 Hz) and 125.90 (d, J_C-F

=

20.4 Hz) (CH_Ar), 127.16 (C_Ar), 127.18 (d, ⁴J_C-F= 2.8 Hz, CH_Ar), 129.9 (d, ³J_C-F= 8.5 Hz, CH_Ar), 131.0 (d,

3

³J_C-F= 8.4 Hz) and 131.1 (d, J_C-F= 8.6 Hz) (CH_Ar), 132.5 (C_Ar), 135.47 and 135.50 (C_Ar), 137.9 and

138.0 (C_Ar), 140.2 and 140.3 (C_Ar), 147.5 (C_Ar), 162.8 (d, ¹J_C-F= 245.8 Hz, C_Ar-F), 163.3 (d, ¹J_C-F= 248.3

Hz) and 163.4 (d, ¹J_C-F= 248.3 Hz) (C_Ar-F), 220.7 (C=O). ¹⁹F NMR (235 MHz, CDCl₃): δ = -113.10 and -

113.07 (CF), -111.45 and -111.40 (CF). IR (ATR, cm⁻¹):

˜ = 3539 (w), 2924 (m), 1733 (s), 1581 (s),

ν

1458 (m), 1403 (m), 1260 (s), 1194 (s), 873 (m), 785 (s). MS (EI, 70 eV): m/z (%) = 458 (100) [M⁺], 459

(28), 373 (17), 360 (11), 334 (12), 55 (15), 41 (12). HRMS (ESI, 70 eV): calcd. for C₃₀H₂₈O₂F₂[M⁺]

458.20519; found 458.20528.

2,4-Di-(3,5-dimethoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 h): light yellow solid (89 mg,

70%), mp = 114

–

115 °C, [α]_D= +52.4° (c 1.00, CHCl₃). ¹H NMR (250 MHz, CDCl₃): δ = 0.93 (s, 3H,

CH₃), 1.32 – 1.41 (m, 1H, aliphatic), 1.51–1.63 (m, 5H, aliphatic), 1.92 – 2.17 (m, 4H, aliphatic), 2.31 –

2.39 (m, 1H, aliphatic), 2.45 – 2.61 (m, 4H, aliphatic), 3.81 (br.s, 12H, 4OCH₃), 5.07 (s, 1H, OH), 6.45 –

6.47 (m, 3H, CH_Ar), 6.51 – 6.53 (m, 1H, CH_Ar), 6.71 (d, ⁴J_H-H= 2.3 Hz, 2H, CH_Ar), 7.31 (br.s, 1H, CH_Ar).

¹³C NMR (63 MHz, CDCl₃): δ = 13.8 (CH₃), 21.5 (CH₂), 26.2 (CH₂), 26.5 (CH₂), 28.2 (CH₂), 31.6

(CH₂), 35.8 (CH₂), 37.9 (CH), 44.3 (CH), 48.0 (C), 50.4 (CH), 55.4 (4OCH₃), 99.5 (CH), 100.1 (CH),

107.3 (2CH_Ar), 107.5 (CH_Ar), 108.0 (CH_Ar), 125.6 (C_Ar), 126.7 (CH_Ar), 128.2 (C_Ar), 132.0 (C_Ar), 135.2

(C_Ar), 137.8 (C_Ar), 140.1 (C_Ar), 147.6 (C_Ar), 160.8 (2C_Ar), 161.55 (C_Ar) 161.58 (C_Ar), 220.8 (C=O). IR

(ATR, cm^-1):

˜ = 3508 (w), 2926 (m), 1733 (s), 1589 (s), 1453 (m), 1336 (m), 1201 (s), 1150 (s), 1058

ν

25

(s), 830 (s). MS (EI, 70 eV): m/z (%) = 542 (100), 544 (9), 543 (49), 271 (18). HRMS (EI, 70 eV): calcd.

for C₃₄H₃₈O₆[M⁺] 542.26629; found 542.26555.

2,4-Di-(p-ethylphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 i): light yellow solid (111 mg, 99%),

mp = 113

–

114 °C, [α]_D= +74.0° (c 1.00, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.93 (s, 3H, CH₃),

1.25 – 1.33 (m, 7 H, 2CH₂CH₃+ aliphatic), 1.50–1.63 (m, 5H, aliphatic), 1.89 – 2.19 (m, 4H, aliphatic),

2.31 – 2.39 (m, 1H, aliphatic), 2.45 – 2.57 (m, 4H, aliphatic), 2.65 – 2.77 (m, 4H, 2CH₂CH₃), 4.88 (s, 1H,

OH), 7.21 – 7.29 (m, 5H, CH_Ar+ CDCl₃), 7.33 – 7.35 (m, 2H, CH_Ar), 7.47 – 7.50 (m, 2H, CH_Ar).

¹³C NMR (75 MHz, CDCl₃): δ = 13.8 (CH₃), 15.3 (CH₃), 15.5 (CH₃), 21.5 (CH₂), 26.2 (CH₂), 26.6

(CH₂), 28.54 (CH₂), 28.56 (CH₂), 28.61 (CH₂), 31.6 (CH₂), 35.8 (CH₂), 37.9 (CH), 44.3 (CH), 48.0 (C),

50.4 (CH), 125.6 (C_Ar), 126.8 (CH_Ar), 128.0 (2CH_Ar), 128.1 (C_Ar), 128.82 (CH_Ar), 128.84 (CH_Ar), 129.2

(2CH_Ar), 129.9 (CH_Ar), 130.3 (CH_Ar), 131.9 (C_Ar), 132.9 (C_Ar), 135.0 (C_Ar), 135.5 (C_Ar), 143.1 (C_Ar), 144.0

(C_Ar), 147.9 (C_Ar), 221.0 (C=O). IR (ATR, cm^-1):

˜ = 3527 (w), 2927 (m), 1737 (s), 1511 (m), 1455 (s),

ν

1399 (m), 1257 (m), 1145 (m), 829 (s). MS (EI, 70 eV): m/z (%) = 478 (100) [M⁺], 479 (55), 152 (12),

151 (13), 133 (19), 57 (10), 55 (12), 41 (12), 36 (10). HRMS (ESI, 70 eV): calcd. for C₃₄H₃₈O₂[M+H]

479.29446; found 479.29408.

2,4-Di-(o-fluorophenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 j): light yellow solid (104 mg, 97%),

1

mp = 118

–

119 °C, [α]_D= +92.2° (c 1.00, CHCl₃). A mixture of diastereomers. H NMR (300 MHz,

CDCl₃): δ = 0.93 and 0.94 (2s, 3H, CH₃), 1.34 – 1.42 (m, 1H, aliphatic), 1.50 – 1.64 (m, 5H, aliphatic),

1.95 – 2.17 (m, 4H, aliphatic), 2.36 – 2.54 (m, 5H, aliphatic), 4.69 and 4.71 (2s, 1H, OH), 7.14 – 7.36 (m,

7H, CH_Ar+ CDCl₃), 7.39 – 7.46 (m, 2H, CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ = 13.8 and 13.9 (CH₃),

21.5 (CH₂), 26.0 and 26.1 (CH₂), 26.4 (CH₂), 27.6 and 28.3 (CH₂), 31.55 and 31.58 (CH₂), 35.8 (CH₂),

37.71 and 37.77 (CH), 44.1 and 44.2 (CH), 47.9 and 48.0 (C), 50.39 and 50.43 (CH), 115.72 and 115.76

(CH_Ar), 116.1 – 116.2 (several signals, CH_Ar), 116.5 and 116.6 (CH_Ar), 120.1 (C_Ar), 121.9 and 122.0 (C_Ar),

2

123.19 (d, J_C-F= 17.7 Hz) and 123.28 (d, J_C-F= 17.4 Hz) (C_Ar), 124.2 – 124.3 (several signals, CH_Ar),

2

124.8 – 124.9 (several signals, CH_Ar), 125.29 (d, J_C-F= 15.8 Hz) and 125.39 (d, J_C-F= 16.0 Hz) (C_Ar),

128.2 (CH_Ar), 129.4 (d, ³J_C-F= 8.2 Hz) and 129.5 (d, ³J_C-F= 7.9 Hz) (CH_Ar), 130.20 (d, ³J_C-F= 7.9 Hz) and

3

130.24 (d, J_C-F= 8.1 Hz) (CH_Ar), 131.9 – 132.3 (several signals, 2CH_Ar+ 2C_Ar), 136.6 and 136.7 (C_Ar),

1

148.3 and 148.4 (C_Ar), 160.0 (d, J_C-F= 246.8 Hz, 2C_Ar-F), 220.9 (C=O). ¹⁹F NMR (235 MHz, CDCl₃):

δ = -112.7 and -112.9 (CF), -114.016 and -114.024 (CF). IR (ATR, cm^-1):

˜ = 3550 (w), 2925 (w), 1731

ν

(m), 1576 (w), 1444 (m), 1414 (m), 1206 (m), 755 (s). MS (EI, 70 eV): m/z (%) = 458 (100) [M⁺], 459

(31), 373 (20), 360 (11), 334 (24), 55(11). HRMS (EI, 70 eV): calcd. for C₃₀H₂₈O₂F₂: 458.20519; found

458.20501.

2,4-Di-(3,5-dimethylphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 k): light yellow solid (111 mg,

1

99%), mp = 111

–

112 °C, [α]_D= +104.1° (c 1.00, CHCl₃). H NMR (300 MHz, CDCl₃): δ = 0.94 (s,

3H, CH₃), 1.36 – 1.64 (m, 6H, aliphatic), 1.90 – 2.17 (m, 4H, aliphatic), 2.36 – 2.37 (m, 13H, 4CH₃+

26

aliphatic), 2.46 – 2.59 (m, 4H, aliphatic), 4.94 (br.s, 1H, OH), 6.92 (s, 1H, CH_Ar), 6.94 (s, 1H, CH_Ar), 6.98

(s, 1H, CH_Ar), 7.05 (s, 1H, CH_Ar), 7.17 (m, 2H, CH_Ar), 7.26 (s, 1H, CH_Ar+ CDCl₃). ¹³C NMR (75 MHz,

CDCl₃): δ = 13.9 (CH₃), 21.39 (2CH₃), 21.41 (2CH₃), 21.6 (CH₂), 26.2 (CH₂), 26.6 (CH₂), 28.5 (CH₂),

31.7 (CH₂), 35.9 (CH₂), 38.0 (CH), 44.4 (CH), 48.0 (C), 50.5 (CH), 125.8 (C_Ar), 126.6 (CH_Ar), 127.0

(2CH_Ar), 127.6 (CH_Ar), 128.0 (CH_Ar), 128.3 (C_Ar), 128.9 (CH_Ar), 129.6 (CH_Ar), 131.8 (C_Ar), 134.8 (C_Ar),

135.9 (C_Ar), 138.0 (2C_Ar), 138.2 (C_Ar), 138.86 (C_Ar), 138.88 (C_Ar), 147.7 (C_Ar), 221.0 (C=O). IR (ATR,

cm⁻¹):

˜ = 3525 (w), 2917 (s), 1737 (s), 1599 (s), 1453 (s), 1203 (m), 1035 (m), 847 (s), 706 (s). MS (EI,

ν

70 eV): m/z (%) = 478 (100) [M⁺], 479 (38). HRMS (ESI, 70 eV): calcd. for C₃₄H₃₈O₂[M⁺] 478.28663;

found 478.28554.

2,4-Di-(p-chlorophenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 l): dark yellow solid (28 mg, 24%),

mp = 119

–

120 °C, [α]_D= +58.2° (c 0.65, CHCl₃). ¹H NMR (300 MHz, CDCl₃): δ = 0.93 (m, 3H, CH₃),

1.32 – 1.36 (m, 1H, ailiphatic), 1.55–1.62 (m, 5H, aliphatic), 1.96 – 2.17 (m, 4H, aliphatic), 2.29 – 2.37

(m, 1H, aliphatic), 2.44 – 2.55 (m, 4H, aliphatic), 4.71 (s, 1H, OH), 7.25 – 7.30 (m, 3H, CH_Ar+ CDCl₃),

7.38 – 7.41 (m, 2H, CH_Ar), 7.46 – 7.54 (m, 4H, CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ = 13.8 (CH₃), 21.5

(CH₂), 26.2 (CH₂), 26.5 (CH₂), 28.6 (CH₂), 31.6 (CH₂), 35.8 (CH₂), 37.8 (CH), 44.2 (CH), 47.9 (C), 50.4

(CH_Ar), 124.7 (C_Ar), 127.1 (CH_Ar), 128.6 (2CH_Ar), 129.7 (2CH_Ar), 130.6 (2CH_Ar), 130.59 (C_Ar), 131.5

(CH_Ar), 131.8 (CH_Ar), 132.5 (C_Ar), 133.2 (C_Ar), 134.1 (C_Ar), 134.3 (C_Ar), 135.4 (C_Ar), 136.5 (C_Ar), 147.54

(C_Ar), 220.8 (C=O). IR (ATR, cm^-1):

˜ = 3533 (w), 2922 (m), 1731 (s), 1490 (s), 1455 (s), 1230 (m), 1087

ν

(s), 1014 (s), 819 (s), 802 (s), 732 (m), 720 (m), 693 (m), 665 (m), 640 (m), 581 (m). MS (EI, 70 eV):

m/z (%) = 490 (100) [M⁺], 494 (14), 493 (21), 492 (69), 491 (34). HRMS (EI, 70 eV): calcd. for

C₃₀H₂₈O₂Cl₂[M+] 490.14609; found 490.14496; calcd. for C₃₀H₂₈O₂Cl³⁷Cl [M+] 492.14314; found

492.14362.

2,4-Di-(2,3,4-trimethoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 m): light yellow solid

(52 mg, 37%), mp = 105

–

106 °C, [α]_D= +43.4° (c 1.00, CHCl₃). A mixture of diastereomers. ¹H NMR

(300 MHz, CDCl₃): δ = 0.94 and 0.95 (2s, 3H, CH₃), 1.54 – 1.62 (m, 6H, aliphatic), 1.91 – 2.17 (m, 4H,

aliphatic), 2.30 – 2.37 (m, 1H, aliphatic), 2.45 – 2.54 (m, 4H, aliphatic), 3.68 – 3.75 (m, 6H, 2OCH₃),

3.89 – 3.93 (m, 12H, 4OCH₃), 6.40 and 6.42 (2s, 1H, OH), 6.76 – 6.89 (m, 3H, CH_Ar), 7.06 – 7.09 (m,

1H, CH_Ar), 7.22 – 7.26 (d, ³J_H-H= 4.5 Hz, 1H, CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ = 13.86 and 13.93

(CH₃), 21.5 (CH₂), 26.0 and 26.2 (CH₂), 26.6 and 27.4 (CH₂), 28.3 and 29.7 (CH₂), 31.6 (CH₂), 35.9

(CH₂), 37.7 and 37.9 (CH), 44.3 (CH), 48.0 (C), 50.5 and 50.6 (CH), 55.9 and 56.1 (2OCH₃), 60.8 – 61.1

(several signals, 3OCH₃), 61.50 and 61.53 (OCH₃), 107.54 and 107.59 (CH_Ar), 108.5 (CH_Ar), 122.8 and

123.0 (C_Ar), 124.0 (C_Ar), 125.30 and 125.33 (CH_Ar), 125.52 and 125.56 (C_Ar), 126.06 and 126.12 (C_Ar),

126.2 (CH_Ar), 127.2 and 127.4 (CH_Ar), 131.9 and 132.1 (C_Ar), 136.3 and 136.5 (C_Ar), 142.2 (C_Ar), 148.9

and 149.0 (C_Ar), 150.34 and 150.36 (C_Ar), 151.5 (C_Ar), 152.0 (C_Ar), 153.05 and 153.14 (C_Ar), 153.2 (C_Ar),

C=O not given. IR (ATR, cm⁻¹):

˜ = 2924 (w), 1734 (m), 1596 (w), 1494 (m), 1455 (s), 1405 (s), 1289

ν

27

(s), 1212 (m), 1093 (s), 1006 (s), 797 (m). MS (EI, 70 eV): m/z (%) = 602 (100) [M⁺], 604 (12), 603 (56),

43 (13). HRMS (EI, 70 eV): calcd. for C₃₆H₄₂O₈[M+] 602.28742; found 602.28714.

2,4-Di-(o-ethoxyphenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 n): dark yellow solid (86 mg, 72%),

1

mp = 94

–

95 °C, [α]_D= +52.4° (c 1.00, CHCl₃). A mixture of diastereomers. H NMR (250 MHz,

CDCl₃): δ = 0.94 and 0.95 (2s, 3H, CH₃), 1.21 – 1.28 (m, 4H, CH₂CH₃+ aliphatic), 1.31 – 1.38 (m, 3H,

CH₂CH₃), 1.49 – 1.64 (m, 5H, aliphatic), 1.89 – 2.17 (m, 4H, aliphatic), 2.31 – 2.59 (m, 5H, aliphatic),

4.00 – 4.14 (m, 4H, 2CH₂CH₃), 6.06 and 6.07 (2s, 1H, OH), 6.99 – 7.20 (m, 5H, CH_Ar), 7.24 – 7.26 (m,

1H, CH_Ar+ CDCl₃), 7.29 – 7.44 (m, 3H, CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ = 13.87 and 13.92 (CH₃),

14.7 (CH₂CH₃), 14.8 and 14.9 (CH₂CH₃), 21.6 (CH₂), 25.95 and 26.09 (CH₂), 26.6 and 26.7 (CH₂), 27.1

and 28.0 (CH₂), 31.7 (CH₂), 35.9 (CH₂), 37.9 (CH), 44.4 (CH), 48.0 (C), 50.58 and 50.62 (CH), 64.04 and

64.2 (CH₂CH₃), 64.98 (CH₂CH₃), 113.12 (CH_Ar), 113.2 and 113.4 (CH_Ar), 120.92 and 120.96 (CH_Ar),

121.93 and 121.96 (CH_Ar), 123.50 and 123.56 (C_Ar), 126.45 and 126.48 (C_Ar), 127.2 and 127.33 (C_Ar),

127.34 and 127.43 (CH_Ar), 128.63 – 128.72 (several signals, 2CH_Ar+ C_Ar), 131.59 – 131.69 (several

signals, CH_Ar+ C_Ar), 132.5 (CH_Ar), 136.0 and 136.2 (C_Ar), 148.97 and 149.10 (C_Ar), 155.01 and 155.05

(C_Ar), 156.16 and 156.55 (C_Ar), 221.1 (C=O). IR (ATR, cm^-1):

˜ = 3371 (w), 2924 (w), 1736 (m), 1596

ν

(w), 1579 (w), 1439 (m), 1227 (m), 1116 (m), 1084 (m), 1038 (m), 750 (s). MS (EI, 70 eV): m/z (%) =

510 (100) [M⁺], 511 (36), 317 (9), 271 (7), 107 (8), 29 (9). HRMS (EI, 70 eV): calcd. for C₃₄H₃₈O₄:

510.27646; found 510.27613.

2,4-Di-(p-fluorophenyl)-3-hydroxyestra-1,3,5(10)-trien-17-one (9 o): yellow solid (89 mg, 75%), mp =

126

–

127 °C, [α]_D= +50.3° (c 1.00, CHCl₃). ¹H NMR (300 MHz, CDCl ): δ = 0.93 (s, 3H, CH₃), 1.50–

3

1.63 (m, 6H, aliphatic), 1.96 – 2.19 (m, 4H, aliphatic), 2.36 (m, 1H, aliphatic), 2.46 – 2.55 (m, 4H,

aliphatic), 4.70 (s, 1H, OH), 7.09 – 7.14 (m, 2H, CH_Ar), 7.21 – 7.27 (m, 5H, CH_Ar+ CDCl₃), 7.50 – 7.54

(m, 2H, CH_Ar). ¹³C NMR (63 MHz, CDCl ): δ = 13.8 (CH₃), 21.5 (CH₂), 26.2 (CH₂), 26.5 (CH₂), 28.6

3

(CH₂), 31.6 (CH₂), 35.8 (CH₂), 37.8 (CH), 44.3 (CH), 47.9 (C), 50.4 (CH), 115.3 (²J_C-F=21.3 Hz, 2CH_Ar),

116.5 (²J_C-F=21.4 Hz, 2CH_Ar), 124.8 (C_Ar), 127.1 (CH_Ar), 127.2 (C_Ar), 130.9 (³J_C-F=7.9 Hz, 2CH_Ar), 131.4

(⁴J_C-F= 3.0 Hz, C_Ar), 131.6 (³J_C-F=8.1 Hz, CH_Ar), 132.1 (³J_C-F=8.0 Hz, CH_Ar), 132.3 (C_Ar), 134.0 (⁴J_C-F

3.1 Hz, C_Ar), 135.3 (C_Ar), 147.7 (C_Ar), 162.1 (¹J_C-F= 245.7 Hz, CF_Ar), 162.5 (¹J_C-F= 248.1 Hz, C_ar-F),

221.1 (C=O). IR (ATR, cm^-1):

= 3540 (w), 2924 (m), 1732 (s), 1599 (w), 1507 (s), 1457 (m), 1217 (s),

=

ν

˜

1156 (m), 831 (s). MS (EI, 70 eV): m/z (%) = 458 (100), 459 (28), 373 (10). HRMS (EI, 70 eV): calcd.

for C₃₀H₂₈O₂F₂: 458.20519; found 458.20512.

General procedure for the synthesis of compounds 10 and 11: 4-(o-fluorophenyl)-estrone (7 k) or

2,4-di-(o-fluorophenyl)-estrone (9 j) (0.274 mmol), K₂CO₃(75.9 mg, 2.0 equiv.) and DMF were added in

28

a round-bottom 50 ml flask and stirred 4 h at 150 °C. The solvent was evaporated in vacuo and the

residue was purified by column chromatography (silica gel, heptane/EtOAc = 3:1).

Benzofuro[2’,3’;3,4]estra-1,3,5(10)-trien-17-one (10): according to the general procedure compound 10

was isolated as a white solid (65 mg, 69%), mp = 268

–

269 °C, [α]_D= +97.6° (c 1.00, CHCl₃). ¹H NMR

(300 MHz, CDCl₃): δ = 0.95 (s, 3H, CH₃), 1.58 – 1.73 (m, 6H, aliphatic), 1.99 – 2.04 (m, 1H, aliphatic),

2.12 – 2.29 (m, 3H, aliphatic), 2.44 – 2.58 (m, 3H, aliphatic), 3.25 – 3.37 (m, 1H, aliphatic), 3.46 – 3.55

(m, 1H, aliphatic), 7.32 – 7.48 (m, 4H, CH_Ar), 7.58 (d, ³J_H-H= 8.0 Hz, 1H, CH_Ar), 8.01 (d, ³J_H-H= 7.7 Hz,

1H, CH_Ar). ¹³C NMR (63 MHz, CDCl₃): δ = 12.9 (CH₃), 20.6 (CH₂), 25.3 (CH₂), 25.5 (CH₂), 26.8 (CH₂),

30.7 (CH₂) 34.9 (CH₂), 37.0 (CH), 43.4 (CH), 46.9 (C), 49.4 (CH), 107.7 (CH_Ar), 110.5 (CH_Ar), 121.3

(C_Ar), 121.5 (CH_Ar), 121.7 (CH_Ar), 123.6 (CH_Ar), 123.9 (C_Ar), 125.4 (CH_Ar), 131.3 (C_Ar), 133.0 (C_Ar),

153.4 (C_Ar), 155.4 (C_Ar), C=O not given. IR (ATR, cm⁻¹):

˜ = 2961 (w), 1733 (w), 1258 (s), 1208 (w),

ν

1012 (s), 799 (s), 746 (s). MS (EI, 70 eV): m/z (%) = 344 (100) [M⁺], 345 (27), 287 (10), 259 (22), 246

(15), 234 (10), 233 (15), 232 (10), 231 (15), 220 (28), 219 (26), 218 (23), 207 (13), 205 (22), 189 (12),

181 (15). HRMS (EI, 70 eV): calcd. for C₂₄H₂₄O₂[M⁺] 344.17708; found 344.17727.

2-(o-Fluorophenyl)-benzofuro[2’,3’;3,4]estra-1,3,5(10)-trien-17-one (11): according to the general

procedure compound 11 was isolated as a light yellow solid (74 mg, 77%), mp = 256

– 257 °C, [α]_D=

1

+96.3° (c 1.00, CHCl₃). H NMR (300 MHz, CDCl₃): δ = 0.96 (s, 3H, CH₃), 1.60 – 1.81 (m, 6H,

aliphatic), 2.05 – 2.32 (m, 4H, aliphatic), 2.51 – 2.60 (m, 3H, aliphatic), 3.30 – 3.42 (m, 1H, aliphatic),

3.52 – 3.60 (m, 1H, aliphatic), 7.29 – 7.47 (m, 5H, CH_Ar), 7.53 – 7.58 (m, 2H, CH_Ar), 7.63 – 7.69 (m, 1H,

CH_Ar), 8.03 – 8.06 (m, 1H, CH_Ar). ¹³C NMR (75 MHz, CDCl₃): δ = 13.9 (CH₃), 21.7 (CH₂), 26.3 (CH₂),

26.6 (CH₂), 27.8 (CH₂), 31.7 (CH₂), 35.9 (CH₂), 38.0 (CH), 44.4 (CH), 47.9 (C), 50.4 (CH), 111.7 (CH_Ar),

116.9 (d, ²J_C-F= 22.4 Hz, CH_Ar), 117.3 (C_Ar), 122.60 (CH_Ar), 122.64 (C_Ar), 122.70 (CH_Ar), 124.1 (d, ⁴J_C-F

=

3.4 Hz, CH_Ar), 124.5 (d, ²J_C-F= 15.0 Hz, C_Ar), 124.9 (C_Ar), 126.0 (d, ⁴J_C-F= 2.5 Hz, CH_Ar), 126.5 (CH_Ar),

129.5 (d, ³J_C-F= 8.3 Hz, CH_Ar), 131.8 (d, ³J_C-F= 3.3 Hz, CH_Ar), 132.2 (C_Ar), 134.2 (C_Ar), 151.8 (C_Ar), 156.4

(C_Ar), 156.8 (d, ¹J_C-F= 245.2 Hz, C_Ar-F), 220.9 (C=O). ¹⁹F NMR (282 MHz, CDCl₃): δ = -114.6 (CF). IR

(ATR, cm⁻¹):

˜ = 2923 (m), 1730 (s), 1445 (m), 1254 (m) 1212 (s), 1010 (m), 749 (s). GC/MS (EI, 70

ν

eV): m/z (%) = 438 (100) [M⁺], 439 (28), 353 (12), 327 (9), 314 (21), 313 (16), 312 (11), 299 (15), 275

(22). HRMS (EI, 70 eV): calcd. for C₃₀H₂₇O₂F [M+] 438.19896; found 438.19875.

Isocoumarino[3’,4’;3,4]estra-1,3,5(10)-trien-17-one (12): 4-Bromoestrone (6 a) (200 mg, 0.574 mmol),

o-ethoxycarbonylphenyl boronic acid (340 mg, 3.0 equiv.), Pd(PPh₃)₄(33.2 mg, 0.05 equiv.), RuPhos

(13.4 mg, 0.05 equiv.), K₂CO₃(318.0 mg, 4.0 equiv.) and dioxane (4 ml) were added in a pressure tube

under argon atmosphere. The mixture was stirred at 110 °C 72 h. The solvent was evaporated in vacuo.

The residue was purified by column chromatography (silica gel, heptane/EtOAc = 2:1). 12 was isolated as

1

a pale pink solid (158 mg, 74%), mp = 235

–

236 °C, [α]_D= +131.4° (c 1.00, CHCl₃). H NMR (300

29

MHz, CDCl₃): δ = 0.97 (s, 3H, CH₃), 1.36 – 1.46 (m, 1H, aliphatic), 1.55 – 1.74 (m, 5H, aliphatic), 1.98 –

2.23 (m, 4H, aliphatic), 2.37 – 2.44 (m, 2H, aliphatic), 2.49 – 2.58 (m, 1H, aliphatic), 3.28 – 3.36 (m, 1H,

aliphatic), 3.41 – 3.52 (m, 1H, aliphatic), 7.21 (d, ³J_H-H= 8.7 Hz, 1H, CH_Ar), 7.46 (d, ³J_H-H= 8.7 Hz, 1H,

CH_Ar), 7.53 – 7.59 (m, 1H, CH_Ar), 7.75 – 7.81 (m, 1H, CH_Ar), 8.35 (d, ³J_H-H= 8.4 Hz, 1H, CH_Ar), 8.46 (dd,

3

⁴J_H-H= 1.5 Hz, J_H-H= 7.8 Hz, 1H, CH_Ar). ¹³C NMR (75 MHz, CDCl₃): δ = 14.0 (CH₃), 21.5 (CH₂),

26.68 (CH₂), 26.75 (CH₂), 31.8 (CH₂), 32.5 (CH₂), 35.9 (CH₂), 37.6 (CH), 45.6 (CH), 48.0 (C), 50.3

(CH), 115.7 (CH_Ar), 117.3 (C_Ar), 122.6 (C_Ar), 127.2 (CH_Ar), 127.92 (CH_Ar), 127.95 (CH_Ar), 130.8 (CH_Ar),

133.8 (CH_Ar), 135.7 (C_Ar), 135.9 (C_Ar), 136.9 (C_Ar), 150.3 (C_Ar), 161.4 (C_Ar), 220.3 (C=O). IR (ATR,

cm⁻¹):

˜ = 2935 (w), 2922 (w), 1714 (s), 1487 (m), 1401 (w), 1256 (s), 1002 (m), 958 (m), 782 (s), 691

ν

(s). GC/MS (EI, 70 eV): m/z (%) = 372 (100) [M⁺], 373 (28), 248 (14), 202 (12), 189 (12), 165 (10).

HRMS (EI, 70 eV): calcd. for C₂₅H₂₄O₃[M+] 372.17200; found 372.17175.

Protocol of Lipase Assay

The lipase inhibitory activity of synthesized derivatives was carried out with the slight modifications in

previously reported method.²⁰Initially, all of the derivatives were tested at the a concentration of 0.1 mM.

The assay buffer was composed of 100 mM Tris-HC1 (pH 7.0) and 5 mM CaCl₂. The enzyme buffer

was prepared by adding 10 mM MOPS (morpholinepropanesulphonic acid) and 1 mM EDTA and the

pH of buffer was adjusted to 6.8. Total assay volume of 200 µL, contained 160 µL assay buffer, 20 µL of

test compound and 10 µL of the pancreatic lipase enzyme (0.5 µg per well). The reaction mixture was

o

allowed to incubate for 10 min at 37 C. Then the absorbance was measured as pre-read at 405 nm by

using micro-plate reader (BioTek^TMELx800, Instruments, Inc. USA). After that the reaction was initiated

by the addition of 10 µL of p-NPB (p-nitrophenylbutyrate) as substrate solution and the absorbance was

measured after 20 min of incubation. Orlistat was used as a positive control. The compounds with

percentage inhibition of above 50 were further dilluted to different concentrations for the determination of

IC₅₀values. All the experiments were performed in triplicate. IC₅₀values were calculated by non-linear

regression analysis of program PRISM 5.0 (GraphPad, San Diego, California, USA).

Acknowledgements.

Financial support by the DAAD (scholarships for A. I. and program Deutsch-Pakistanische

Hochschulzusammenarbeit) is gratefully acknowledged. J. Iqbal is thankful to the Organization for the

Prohibition of Chemical Weapons (OPCW), The Hague, The Netherlands and Higher Education

Commission of Pakistan for the financial support through Project No. 20-3733/NRPU/R&D/ 14/520 for

the financial support.

Article Doi

Synthesis, functionalization and biological activity of arylated derivatives of (+)-estrone

DOI: 10.1016/j.bmc.2016.12.009

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Article abstract of DOI:10.1016/j.bmc.2016.12.009

Full text of DOI:10.1016/j.bmc.2016.12.009

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