Supporting Information
Organogelation and cytotoxic evolution of phosphonate ester functionalised
hydrophobic alkanediamide motifs
Kamalakar P. Nandre,a,d Sidhanath V. Bhosale,a,* Rajesh S. Bhosale,a
Sujitha Pombala,b C. Ganesh Kumar,b Kay Lathamc and Sheshanath V.
Bhosalec,*
a
Polymers and Functional Materials Division, CSIR-Indian Institute of Chemical
Technology, Hyderabad-500607, Andra Pradesh, India.
b
Chemical Biology Laboratory, Natural Product Division, CSIR-Indian Institute of
Chemical Technology, Hyderabad 500607, India.
c
School of Applied Sciences, RMIT University, GPO Box 2476, Melbourne, Vic-3001,
Australia.
d
Department of Organic Chemistry, School of Chemical Sciences, North Maharashtra
University, Jalgaon-425001, India
*Corresponding
author.
Email:
bhosale@iict.res.in
sheshanath.bhosale@rmit.edu.au (Shesh. V. Bhosale).
(Sid.
V.
Bhosale);
Table of Contents
1. Materials, instruments and methods ----------------------------------------------------- S2
2. Synthesis and their molecular Identification ------------------------------------------- S3
3. Microscopic analysis, for example: UV-Vis absorption, fluorescence, SEM, TEM,
TGA, DTG, DSC and FTIR -------------------------------------------------------------- S7
4.
1
H NMR and 13C NMR Spectra’s of new compounds ------------------------------- S12
5. References --------------------------------------------------------------------------------- S27
1. Methods, Materials and Instrumentation:
S1
All reagents were purchased either from Sigma Aldrich Chemical Co. or Merck and used as
such without any further purification. All the solvents were received from commercial
sources and purified by standard methods. For all aqueous spectroscopic studies, water of
spectroscopy or HPLC grade was used. Melting points were determined in capillaries and are
uncorrected.1H NMR spectra were recorded on ADVANCE 300MHz and INNOVA 500
MHz NMR spectrometer and all the spectra were calibrated against TMS. Mass spectrometric
data were obtained by electron spray ionization (ESI-MS) technique on an Agilent
Technologies 1100 Series (Agilent Chemistation Software) mass spectrometer. Highresolution mass spectra (HRMS) were obtained by using ESI-QTOF mass spectrometry.
FTIR spectra were obtained on a Perkin Elmer FT-IR 400 spectrometer.
(a) Nuclear magnetic resonance (NMR) spectroscopy: All 1H NMR and 13C spectra were
recorded on AVANCE 300 MHz and INNOVA 500 MHz NMR spectrometers and
referenced to TMS.
(b) Mass and HRMS spectroscopy: Mass spectrometric data were obtained using the
electron spray ionization (ESI-MS) technique on an Agilent Technologies 1100 Series mass
spectrometer. High-resolution mass spectra (HRMS) were obtained by using ESI-Q-TOF
mass spectrometry.
(c) UV-Visible Measurements: All UV-Visible spectra were recorded in a Shimadzu 2450
UV-visible spectrometer in 1cm path length cuvette. The solutions were allowed to
equilibrate at room temp for 2 h before spectral measurements.
(d) Fluorescence Measurements: Fluorescence emission spectra were recorded in a
Horiba Jobin Yvon FluoroMax®-4 –Spectrofluorometer. All experiments were performed in
a quartz cell with a 1 cm path length with 240 nm excitation wavelength.
S2
(e) Field Emission Scanning Electron Microscopy (FE-SEM): FE-SEM measurements
were performed on an electron microscopy HITACHI S-4800, operating at an accelerating
voltage of 15 kV. Freshly prepared dispersion of organogels or xerogels was dropped onto a
carbon tape and the solvent was allowed to evaporate before investigation by SEM and
images were collected.
(f) Differential scanning calorimetry (DSC): Differential scanning calorimetry was
carried out by using a DSC Q 100. 20 mg of compound was placed into a large volume
capsule (LVC) that was then sealed. The sample LVC pan was placed into the DSC apparatus
together with an empty LVC pan as reference. Heating scan was then recorded from 0–200
°C at a scan rate of 1°C min-1.
(g) Thermal Gravimetric Analysis (TGA): Thermo-gravimetric analysis (TGA) of
xerogels was carried out by using a TGA Q500 thermal analyzer. Measurements were carried
out in platinum pans under nitrogen atmosphere (flow rate of 50 mL/ min) with a heating rate
of 10 °C/min over a temperature range of 0 -700 °C.
(h) Fourier Transform Infrared (FTIR) Spectroscopy Measurement: All FTIR
spectra’s were collected on a Perkin Elmer FT-IR 400. The instrument was continuously
purged with CO2-free dry air and spectra’s were recorded between 450 and 4000 cm-1.
2. Molecular Identification
2.1: Precursor synthesis: Three distinct diethyl phosphonate anilines diethyl 4-aminophenyl
phosphonate (A), diethyl (4-aminobenzyl) phosphonate (B) and tetraethyl 1-aminobenzene3,5-bis(methylphosphonate) (C) were exploited for the synthesis of targeted compounds and
prepared by previously reported methodesS1-3.
S3
2.2: General procedure for the preparation of acid chlorides α,ω-diacid dichloride’s: All
acid chlorides were prepared according to the procedure described by Hussein et al.4 All of
the glassware used in the assembly was oven-dried at 120 oC before use. Dicarboxylic acids
were refluxed in thionyl chloride (3 mL) and 1 drop DMF in a round-bottomed flask for three
hours. Co-evaporation of the resulting reaction mixture with toluene (3 x 5 mL) until dryness
gave the corresponding acid chlorides as colourless oil.
2.3: Molecular Identification of newly synthesized compounds
Compound (1a): Yield: 61%, mp 172-174 ºC; 1H NMR (300 MHz, CDCl3) : 1.29 (t, J = 6.9
Hz, 12H, 4-CH3CH2O), 1.74- 1.76 (m, 4H, 2-CH2CH2CO), 2.34-2.45 (m, 4H, 2-CH2CO),
4.01- 4.15 (m, 8H, 4-CH2O), 7.67- 7.78 (m, 8H, ArH), 9.16 (s, 2H, NH); 13C NMR (75 MHz,
CDCl3) : 16.1, 24.6, 36.7, 62.2, 119.2, 123.2, 132.5, 143.0, 172.2; IR (KBr,  cm-1): 1029,
1229, 1530, 1594, 1700, 2981, and 3263; ESI-MS (m/z %): 569 (97) [M + H]+, 591(100) [M
+ Na]+; HRMS: calculated for C26H39O8N2P2 = 569.2176; found =569.2187.
Compound (1b): Yield: 67%, mp 187-189 ºC; 1H NMR (500 MHz, CDCl3) : 1.30 (t, J = 7
Hz, 12H, 4-CH3CH2O), 1.66-1.73 (m, 6H, 3-CH2), 2.35 (t, J = 7.5 Hz, 4H, 2-CH2CO), 4.044.14 (m, 8H, 4-CH2O), 7.70-7.72 (m, 8H, ArH), 8.51 (s, 2H, NH);
13
C NMR (75 MHz,
CDCl3) : 16.2, 25.1, 28.8, 37.2, 62.1, 119.2, 119.4, 132.6, 142.6, 172.5; IR (KBr,  cm-1):
1026, 1232, 1527, 1593, 1697, 2927, 3256; ESI-MS (m/z %): 583(100) [M + H]+; HRMS:
calculated for C27H41O8N2P2 = 583.2332; found = 583.2343.
S4
Compound (1c): Yield: 65%, mp 182-184 ºC; 1H NMR (500 MHz, CDCl3) : 1.22-1.28 (m,
4H, 2-CH2), 1.30 (t, J = 7 Hz, 12H, 4-CH3CH2O), 1.61-1.69 (m, 4H, 2-CH2CH2CO), 2.32 (t,
J = 8.5 Hz, 4H, 2-CH2CO), 4.04-4.15 (m, 8H, 4-CH2O), 7.69-7.79 (m, 8H, ArH), 9.26 (s, 2H,
NH); 13C NMR (75 MHz, CDCl3) : 16.2, 25.4, 28.8, 37.2, 62.1, 119.2, 123.1, 132.6, 142.9,
172.8; IR (KBr,  cm-1): 1028, 1231, 1594, 1706, 2981, 3259; ESI-MS (m/z %): 597 (70) [M
+ H]+, 619 (100) [M + Na]+; HRMS: calculated for C28H43O8N2P2 = 597.2489; found =
597.2501.
Compound (1d): Yield: 51%, mp 110-112 ºC; 1H NMR (500 MHz, CDCl3) : 1.20-1.26 (m,
6H, 3-CH2), 1.29 (t, J = 7 Hz, 12H, 4-CH3CH2O), 1.58-1.69 (m, 4H, 2-CH2CH2CO), 2.34 (t,
J = 7 Hz, 4H, 2-CH2CO), 4.02- 4.14 (m, 8H, 4-CH2O), 7.69-7.76 (m, 8H, ArH), 9.15 (s, 2H,
NH);
C NMR (75 MHz, CDCl3) : 16.1, 25.4, 28.9, 28.9, 37.4, 62.1, 119.3, 123.0, 132.4,
13
142.9, 172.9; IR (KBr,  cm-1): 1027, 1229, 1522, 1591, 1700, 2930, 3267; ESI-MS (m/z %):
611 (100) [M + H]+, 634 (67) [M + H + Na]+; HRMS: calculated for C29H45O8N2P2 =
611.2645; found = 611.2657.
Compound (1e): Yield: 72%, mp 162-164 ºC; 1H NMR (300 MHz, CDCl3) : 1.20-1.25 (m,
8H, 4-CH2), 1.29 (t, J = 7.2 Hz, 12H, 4-CH3CH2O), 1.63-1.67 (m, 4H, 2-CH2CH2CO), 2.34
(t, J = 7.5 Hz, 4H, 2-CH2CO), 4.01-4.15 (m, 8H, 4-CH2O), 7.67-7.79 (m, 8H, ArH), 9.10 (s,
2H, NH);
13
C NMR (75 MHz, CDCl3) : 16.1, 25.4, 28.9, 37.3, 62.16, 119.2, 123.0, 132.5,
142.8, 172.9; IR (KBr,  cm-1): 1027, 1231, 1528, 1593, 1707, 2982, 3255; ESI-MS (m/z %):
625 (100) [M + H]+, 647 (80) [M + H + Na]+; HRMS: calculated for C30H47O8N2P2 =
625.2807; found = 625.2820.
Compound (1f): Yield: 77%, mp 134-136 ºC; 1H NMR (300 MHz, CDCl3) : 1.19-1.25 (m,
12H, 6-CH2), 1.29 (t, J = 7.5 Hz, 12H, 4-CH3CH2O), 1.61-1.71 (m, 4H, 2-CH2CH2CO), 2.35
(t, J = 7.8 Hz, 4H, 2-CH2CO), 4.01-4.15 (m, 8H, 4-CH2O), 7.67-7.78 (m, 8H, ArH), 8.96 (s,
S5
2H, NH);
13
C NMR (75 MHz, CDCl3) : 16.2, 25.4, 28.9, 29.0, 37.4, 62.1, 119.0, 123.2,
132.5, 142.7, 172.7; IR (KBr,  cm-1): 1029, 1231, 1523, 1592, 1708, 2925, 3265; ESI-MS
(m/z %): 653 (100) [M + H]+, 676 (63) [M + Na]+; HRMS: calculated for C32H51O8N2P2 =
653.3115, found = 653.3131.
Compound (1g): Yield: 75%, mp 98-100 ºC; 1H NMR (300 MHz, CDCl3) : 1.21-1.26 (m,
12H, 6-CH2), 1.29 (t, J = 7.5 Hz, 12H, 4-CH3CH2O), 1.6 (m, 6H, 3-CH2), 2.35 (t, J = 7.5 Hz,
4H, 2-CH2CO), 4.01-4.15 (m, 8H, 4-CH2O), 7.75-7.71 (m, 8H, ArH), 8.6 (s, 2H, NH);
13
C
NMR (75 MHz, CDCl3) : 16.1, 25.3, 28.9, 29.0, 37.4, 62.1, 119.0, 132.5, 132.5, 142.7,
172.7; IR (KBr,  cm-1): 1036, 1230, 1529, 1694, 2929, 3260; ESI-MS (m/z %): 668 (40) [M
+ 2H]+, 690 (100) [M + H + Na]+; HRMS: calculated for C33H53O8N2P2 = 667.3271, found =
667.3288.
Compound (1h): Yield: 73%, mp 115-117 ºC; 1H NMR (300 MHz, CDCl3) : 1.20-1.26 (m,
20H, 10-CH2), 1.28 (t, J = 7.0 Hz, 12H, 4-CH3CH2O), 1.71-1.74 (m, 4H, 2-CH2CH2CO),
2.37 (t, J = 7.5 Hz, 4H, 2-CH2CO), 4.00-4.17 (m, 8H, 4-CH2O), 7.70-7.76 (m, 8H, ArH), 8.37
(s, 2H, NH);
13
C NMR (75 MHz, CDCl3) : 16.2, 25.4, 28.0, 29.2, 37.5, 62.0, 119.1, 123.4,
132.8, 142.4, 172.3; IR (KBr,  cm-1): 1027, 1221, 1526, 1592, 1694, 2982, 3248; ESI-MS
(m/z %): 710 (50) [M + H]+, 732 (100) [M + H + Na]+; HRMS: calculated for
C36H58O8N2NaP2 = 731.3560; found = 731.3595.
Compound (2a): Yield: 78%, mp 172-174 ºC; 1H NMR (300 MHz, CDCl3) : 1.23 (t, J = 6.9
Hz, 12 H, 4-CH3CH2O), 1.31 (m, 8H, 4-CH2), 2.29 (t, J = 7.2 Hz, 4H, 2-CH2CO), 3.06 (d, JHP
= 21.3 Hz, 4H, 2-CH2Ar), 3.96-4.06 (m, 8H, 4-CH2O), 7.17 (d, J = 6.6, 4H, ArH), 7.46 (d, J
= 8.1 Hz, 4H, ArH), 8.24 (s, 2H, NH); 13C NMR (75 MHz, CDCl3) : 16.3, 25.4, 28.9, 31.9,
33.7, 37.2, 62.1, 119.8, 126.08, 129.98, 137.66, 172.16; IR (KBr,  cm-1): 1024, 1231, 1540,
S6
1606, 1680, 2943, 3259; ESI-MS (m/z %): 625 (100) [M + H]+; HRMS: calculated for
C30H47O8N2P2 = 625.2807; found = 625.2820.
Compound (2b): Yield: 80%, mp 204-206 ºC, 1H NMR (300 MHz, CDCl3) : 1.23 (t, J = 7.8
Hz, 12 H, 4-CH3CH2O), 1.36 (m, 4H, 2-CH2), 1.70 (m, 8H, 4-CH2), 2.30 (t, J = 7.8 Hz, 4H,
2-CH2CO), 3.07 (d, JHP = 21.3 Hz, 4H, 2-CH2Ar), 3.99-4.04 (m, 8H, 4-CH2O), 7.18 (d, J =
6.6, 4H, ArH), 7.47 (d, J = 8.4 Hz, 4H, ArH), 8.27 (s, 2H, NH); 13C NMR (75 MHz, CDCl3)
: 16.3, 25.4, 28.9, 31.9, 33.7, 37.2, 62.1, 119.8, 126.1, 129.9, 137.5, 172.1; IR (KBr,  cm-1):
1024, 1230, 1541, 1605, 1680; ESI-MS (m/z %): 654 (100) [M + H]+; HRMS: calculated for
C32H51O8N2P2 = 653.3091; found = 653.3132.
Compound (3a): Thick yellow oil, Yield: 67%; 1H NMR (300 MHz, CDCl3) : 1.23-1.32 (m,
36 H, 8-CH3CH2O & 6-CH2), 2.30 (t, J = 7.5 Hz, 4H, 2-CH2CO), 2.88 (d, JHP = 22.2 Hz, 8H,
4-CH2Ar), 3.97-4.06 (m, 16H, 8-CH2O), 6.94 (s, 2H, ArH), 7.46-7.51 (m, 4H, ArH), 8.01 (bs,
2H, NH); IR (KBr,  cm-1):1022, 1252, 1603, 1681, 2925, 2982, 3275; ESI-MS (m/z %): 953
(28) [M + H]+, 975 (100) [M + Na]+;
13
C NMR (75 MHz, CDCl3)δ: 16.1, 20.9, 24.7, 25.3,
29.0, 32.3, 34.1, 37.5, 62.2, 119.3, 126.1, 132.0, 138.9, 171.9; ESI-MS (m/z %): 953 (28) [M
+ H]+, 975 (100) [M + Na]+; HRMS: calculated for C42H72O14N2NaP4 = 975.3917; found =
975.3931.
Compound (3b): Thick yellow oil, Yield: 73%, 1H NMR (300 MHz, CDCl3) : 1.22-1.29 (m,
36 H, 8-CH3CH2O & 6-CH2), 1.65-1.72 (m, 4H, 2-CH2CH2CO), 2.29 (t, J = 7.8 Hz, 4H, 2CH2CO), 3.05 (d, JHP = 21.9 Hz, 8H, 4-CH2Ar), 3.97-4.06 (m, 16H, 8-CH2O), 6.92 (s, 2H,
ArH), 7.48-7.55 (m, 4H, ArH), 8.06 (bs, 2H, NH); 13C NMR (75 MHz, CDCl3) : 16.1, 25.4,
29.0, 29.1, 29.5, 32.3, 34.1, 37.3, 62.2, 119.3, 126.0, 132.0, 138.9, 171.9; IR (KBr,  cm1
):1028, 1241, 1563, 1610, 1686, 2927, 2982, 3275; ESI-MS (m/z %): 982 (12) [M + 2H]+,
S7
1004 [M + H + Na]+; HRMS: calculated for C44H76O14N2NaP4 = 1003.4139; found =
1003.4171.
Compound (3c): Thick yellow oil, Yield: 68%, 1H NMR (300 MHz, CDCl3) : 1.23-1.27 (m,
38 H, 8-CH3CH2O & 7-CH2), 1.67-1.69 (m, 4H, 2-CH2CH2CO), 2.20 (t, J = 7.5 Hz, 4H, 2CH2CO), 3.06 (d, JHP = 21.6 Hz, 8H, 4-CH2Ar), 3.97-4.11 (m, 16H, 8-CH2O), 6.92 (s, 2H,
ArH), 7.49- 7.57 (m, 4H, ArH), 8.07 (bs, 2H, NH); 13C NMR (75 MHz, CDCl3) : 16.2, 25.4,
28.8, 29.1, 29.2, 32.3, 34.1, 37.4, 62.3, 119.4, 126.2, 132.0, 138.9, 172.9; IR (KBr,  cm-1):
1040, 1241, 1562, 1609, 1687, 2927, 2982, 3272; ESI-MS (m/z %): 995 (100) [M=Na]+, 1017
(62); HRMS: calculated for C45H78O14N2NaP4 = 1017.4295; found = 1017.4329.
Compound (3d): Thick yellow oil, Yield: 71%, 1H NMR (300 MHz, CDCl3) : 1.22-1.27 (m,
40H, 8-CH3CH2O & 8-CH2), 1.65-1.69 (m, 4H, 2-CH2CH2CO), 2.29 (t, J = 7.8 Hz, 4H, 2CH2CO), 3.05 (d, JHP = 21.6 Hz, 8H, 4-CH2Ar), 3.96-4.06 (m, 16H, 8-CH2O), 6.92 (s, 2H,
ArH), 7.48-7.53 (m, 4H, ArH), 8.23 (s, 2H, NH); 13C NMR (75 MHz, CDCl3) : 16.2, 25.4,
29.1, 29.2, 32.4, 34.2, 37.4, 62.2, 119.5, 126.2, 132.1, 138.8, 171.8; IR (KBr,  cm-1): 1040,
1241, 1562, 1609, 1687, 2927, 2982, 3273; ESI-MS (m/z %): 1010 (10) [M + H]+, 1032 (20)
[M + Na]+; HRMS: calculated for C46H80O14N2NaP4 = 1031.4452; found = 1031.4483.
Compound (3e): Thick yellow oil, Yield:78%, 1H NMR (500 MHz, CDCl3) : 1.23-1.33 (m,
44 H, 8-CH3CH2O & 10-CH2), 1.66-1.72 (m, 4H, 2-CH2CH2CO), 2.30 (t, J = 8.0 Hz, 4H, 2CH2CO), 3.07 (d, JHP = 21.5 Hz, 8H, 4-CH2Ar), 3.99-4.04 (m, 16H, 8-CH2O), 6.93 (s, 2H,
ArH), 7.46 (s, 4H, NH);
13
C NMR (75 MHz, CDCl3) : 16.2, 25.4, 29.2, 29.2, 29.4, 32.4,
34.2, 37.4, 62.1, 119.4, 126.2, 132.1, 138.8, 171.7; IR (KBr,  cm-1): 1040, 1241, 1562, 1609,
1682, 2925, 2982, 3274; ESI-MS (m/z %): 1038 [M + 2H]+, 1061 (15) [M + 2H + Na]+.
HRMS: calculated for C48H84O14N2NaP4 = 1059.4770; found = 1059.4808.
S8
3. Microscopic analysis
Figure S1: The plot of absorption maxima of 1h and 3e versus varying fraction weight of
cyclohexane at 257 nm absorbance.
Figure S2: The plot of emission intensity of 1h and 3e versus varying fraction weight of
cyclohexane at 303 and 344 nm (λext.= 258 nm).
S9
Figure S3: FE-SEM images of glycerol gel: 1e (a), 1f (b), 1h (c); dioxane: cyclohexane (1:6)
xerogel of 1e (d), 1h (e, f); partial dioxane gel (1:7) of 1f (g, h).
S10
Figure S4: FE-SEM images of precipitate solid of 1h from CHCl3: Cyclohexane (1:7)
Figure S5: FE-SEM images for Chloroform: cyclohexane gel (1:7) of 3e (a, b) and of 3d (c,
d).
S11
Figure S6: TGA/DTG thermograph of 1h
Figure S7: TGA/DTG thermograph of 1e
S12
Figure S8: TGA/DTG thermograph of 3e
Figure S9: Differential scanning calorimetry (DSC) of 1h
S13
Figure S10: Differential scanning calorimetry (DSC) of 3e
4. 1H NMR and 13C NMR Spectra’s of new compounds
1H
spectra of compound 1a (CDCl3)
13C
spectra of compound 1a (CDCl3)
S14
1H
spectra of compound 1b (CDCl3)
S15
13C
1H
spectra of compound 1b (CDCl3)
spectra of compound 1c (CDCl3)
S16
13C
1H
spectra of compound 1c (CDCl3)
spectra of compound 1d (CDCl3)
S17
13C
1H
spectra of compound 1d (CDCl3)
spectra of compound 1e (CDCl3)
S18
13C
1H
spectra of compound 1e (CDCl3)
spectra of compound 1f (CDCl3)
S19
13C
1H
spectra of compound 1f (CDCl3)
spectra of compound 1g (CDCl3)
S20
13C
1H
spectra of compound 1g (CDCl3)
spectra of compound 1h (CDCl3)
S21
13C
1H
spectra of compound 1h (CDCl3)
spectra of compound 2a (CDCl3)
S22
13C
1H
spectra of compound 2a (CDCl3)
spectra of compound 2b (CDCl3)
S23
13C
1H
spectra of compound 2b (CDCl3)
spectra of compound 3a (CDCl3)
S24
13C
1H
spectra of compound 3a (CDCl3)
spectra of compound 3b (CDCl3)
S25
13C
1H
spectra of compound 3b (CDCl3)
spectra of compound 3c (CDCl3)
S26
13C
1H
spectra of compound 3c (CDCl3)
spectra of compound 3d (CDCl3)
S27
13C
1H
spectra of compound 3d (CDCl3)
spectra of compound 3e (CDCl3)
S28
13C
spectra of compound 3e (CDCl3)
4. References:
1. R. J. Cooper, P. J. Camp, R. J. Gordon, D. K. Henderson, D. C. R. Henry, H. McNab,
S. S. De Silva, D. Tackley, P. A. Tasker and P. Wight, Dalton Trans., 2006, 2785.
2. E. A. Wydysh, S. M. Medghalchi, A. Vadlamudi and C. A. Townsend, J. Med. Chem.
2009, 52, 3317.
3. Y. C. Kim, S. G. Brown, T. K. Harden, J. L .Boyer, G. Dubyak, B. F. King, G.
Burnstock and K. A. Jacobson, J. Med. Chem. 2001, 44, 340.
S29