Gloria Uccello-Barretta,* Federica Balzano, Federica Aiello, Francesca Nardelli, Alessia Ciogli, Andrea
Calcaterra and Bruno Botta
Supporting Information
Experimental
1 H NMR signals of hosts 1 4 and guest 6 .
Tables S1–S9
Figures S1–S4
Schemes S1-S2
Experimental
For MS measurements, the employed conditions are: source voltage = +5.0 kV, sheath gas (N
2
) = 25 AU
(Arbitrary Units), auxiliary gas = 0 AU, capillary voltage = +3.0 V, capillary temperature = 548 K, tube lens offset = -15 V. HRMS were obtained with an APEX III (7 T Magnet) FTICR mass spectrometer equipped with an Apollo ESI source (Bruker Daltonics).
NMR measurements for the characterization of hosts 1 5 and for interaction studies were performed on a spectrometer operating at 600 MHz and 150 MHz for 1 H and 13 C nuclei respectively. The temperature was controlled to ±0.1 °C. All 1 H NMR chemical shifts are referenced to TMS as external standard. The 2D NMR spectra were obtained by using standard sequences with the minimum spectral width required. Proton 2D gCOSY
(gradient COrrelated SpectroscopY) spectra were recorded with 128 increments of 2 scans and 2K data points.
The relaxation delay was 3 s. 2D TOCSY (TOtal Correlation SpectroscopY) spectra were recorded by employing mixing times of 80 ms and 120 ms. The pulse delay was maintained at 3 s; 256 increments of 4 scans and 2K data points each were collected. The 2D ROESY (Rotating-frame Overhauser Enhancement SpectroscopY) experiments were performed by employing a mixing time of 0.3 s or 0.6 s. The pulse delay was maintained at 5 s;
256 increments of 6 scans and 2K data points each were collected. Proton 1D ROESY spectra were recorded using the selective pulse SEDUCE generated by means of the Agilent Pandora Software. The selective 1D
ROESY spectra were acquired with 1024 or 512 scans in 32K data points with a 5 s relaxation delay and a mixing time of 0.3 s or 0.6 s. gHSQC (gradient Heteronuclear Single Quantum Coherence) and HMBC
(Heteronuclear Multiple Bond Correlation) spectra were recorded by employing a pulse delay of 1 s and 128 or
200 increments of 64-128 scans. HMBC experiments were optimized for a long-range 1 H13 C coupling constant of 8 Hz. DOSY (Diffusion Ordered SpectroscopY) experiments were carried out by using a stimulated echo sequence with self-compensating gradient schemes, a spectral width of 6000 Hz and 64K data points. Typically, a value of 50 ms was used for

, 2 ms for

, and g was varied in 15 steps (4 transients each) to obtain an approximately 90–95% decrease in the resonance intensity at the largest gradient amplitude. The baselines of all arrayed spectra were corrected prior to processing the data. After data acquisition, each FID was apodized with
1.0 Hz line broadening and Fourier transformed. The data were processed with the DOSY macro (involving the determination of the resonance heights of all the signals above a pre-established threshold and the fitting of the decay curve for each resonance to a Gaussian function) to obtain pseudo two-dimensional spectra with NMR chemical shifts along one axis and calculated diffusion coefficients along the other. Gradient amplitudes in
DOSY experiments have been calibrated by using a standard sample of H
2
O 1%.

(R,R)-1
H
O
NH
H
H
7
6
1
O
NH 2
3
H 4
5t H H
5c
1 H NMR (600 MHz, CDCl
3
, 10 mM, 298 K):

6.20 (dd, J
5t-4
= 17.1 Hz, J
5t-5c
= 1.4 Hz, 2H, H5t), 6.15 (s, 2H, H2), 6.04
(dd, J
4-5t
= 17.1 Hz, J
4-5c
= 10.3 Hz, 2H, H4), 5.60 (dd, J
5c-4
= 10.3 Hz, J
5c-5t
= 1.4 Hz, 2H, H5c), 3.78 (m, 2H, H1), 2.09 (m,
2H, H6eq), 1.79 (m, 2H, H7eq), 1.35 (m, 2H, H7ax), 1.30 (m, 2H, H6ax).
13 C NMR (150 MHz, CDCl
3
, 10 mM, 298 K):

= 166.3 (C3), 130.9 (C4), 126.4 (C5), 54.0 (C1), 32.2 (C6), 24.7 (C7).
(S,S)-2
5c
H H
5t
4 H
3
2 HN
O
1
6
7
8
HN
9
O
1 H NMR (600 MHz, CDCl
3
, 10 mM, 298 K):

= 7.20 (m, 4H, H8), 7.19 (m, 2H, H9), 7.17 (m, 4H, H7), 6.82 (s, 2H, H2),
6.27 (dd, J
5t-4
= 17.0 Hz, J
5t-5c
= 1.3 Hz, 2H, H5 t
), 6.10 (dd, J
4-5t
= 17.0 Hz, J
4-5c
= 10.4 Hz, 2H, H4), 5.65 (dd, J
5c-4
= 10.4 Hz,
J
5c-5t
= 1.3 Hz, 2H, H5 c
), 5.39 (m, 2H, H1).
13 C NMR (150 MHz, CDCl
3
, 10 mM, 298 K):

= 166.3 (C3), 138.4 (C6), 130.6 (C4), 128.7 (C8), 128.0 (C9), 127.6 (C7),
127.1 (C5), 59.6 (C1)
(R,R)-3
10' 12'
39
38
40
34 HN
37
33
32
31 HN
O
6'
4'
O
35
5
30
O
6
4
O
36/36'
B
7
3
29/29'
8
2
O
10
28
O
24
9
1
11
A'
12
O
41/41'
O
42
27
13
14
26
15
19
B'
O
16
17
16'
18
O
18'
25
A
23
21
22
O
20
48/48'
47
O
NH
44
45
43
NH 46
49
52
50
51
24' 22'
1 H NMR (600 MHz, CDCl
3
, 7 mM, 298 K):

= 6.92 (s, 2H, H26/H28), 6.82 (d, J
31-32
= 6.2 Hz, 2H, H31/H43), 6.49 (s, 2H,
H11/H23), 6.14 (s, 2H, H5/H17), 5.97 (s, 2H, H25/H27), 5.51 (d, J
34-33
= 8.9 Hz, 2H, H34/H46), 5.06 (dd, J
2-29’
= 12.4 Hz, J
2-
29
= 5.2 Hz, 2H, H2/H14), 4.60 (dd, J
8-40’
= 13.6 Hz, J
8-40
= 4.1 Hz, 2H, H8/H20), 3.94 (s, 6H, H10’/H22’), 3.91 (s, 6H,
H12’/H24’),3.89 (m, 2H, H33/H45), 3.53 (s, 6H, H4’/H16’), 3.28 (dd, J
29-29’
= 14.3 Hz, J
29-2
= 5.2 Hz, 2H, H29/H41), 3.25 (s,
6H, H6’/H18’), 3.00 (m, 2H, H32/H44), 2.94 (dd, J
36-36’
= 17.0 Hz, J
36-8
= 4.1 Hz, 2H, H36/H48), 2.69 (dd, J
36’-36
= 17.0 Hz,
J
36’-8
= 13.6 Hz, 2H, H36’/H48’), 2.23 (dd, J
29’-29
= 14.3 Hz, J
29’-2
= 12.4 Hz, 2H, H29’/H41’), 2.03 (m, 2H, H37eq/H49eq),

1.82 (m, 2H, H40eq/H52eq), 1.72 (m, 2H, H39eq/H51eq), 1.60 (m, 2H, H38eq/H50eq), 1.22 (m, 2H, H39ax/H51ax), 1.21
(m, 2H, H40ax/H52ax), 1.18 (m, 2H, H38ax/H50ax), 1.11 (m, 2H, H37ax/H49ax).
13 C NMR (150 MHz, CDCl
3
, 10 mM, 298 K):

= 174.4 (C35/C47), 172.2 (C30/C42), 157.2 (C4/C16), 156.3 (C6/C18),
156.0 (C12/C24), 154.8 (C10/C22), 126.5 (C25/C27), 126.1 (C26/C28), 125.1 (C9/C21), 124.2 (C3/C15, C1/C13), 121.3
(C7/C19), 97.5 (C5/C17), 94.5 (C11/C23), 55.9 (C4’/C16’, C10’/C22’), 55.7 (C6’/C18’), 55.3 (C12’/C24’), 57.0 (C32/C44),
51.2 (C33/C45), 41.6 (C36/C48), 40.0 (C29/C41), 33.5 (C2/C14, C8/C20), 32.5 (C37/C49), 31.5 (C40/C52), 25.1
(C39/C51), 24.1 (C38/C50).
(S,S)-4
44
40
43
37
39
38
34
HN
42
41 33
32
HN
31
10' 12'
O
6'
O
35
5
4'
30
4
O
6
O
36/36'
B
7
3
8
2
O
10
28
29/29'
O
24
9
1
11
12
O
45/45' 46
O
A'
27
13
25
A
23
14
15
O
16
16'
26 B' 17
19
21
22
O
20
52/52'
18
18'
O
51
O
NH
48
49
47
NH 50
53
57
54
58
56
55
59
60
24' 22'
1 H NMR (600 MHz, CDCl
3
, 3 mM, 298 K):

= 7.62 (d, J
31-32
= 7.5 Hz, 2H, H31/H47), 7.21-7.18, 7.09-7.00 (m, 20H,
H38/H54, H39/H55, H40/H56, H42/H58, H43/H59, H44/H60), 7.08 (s, 2H, H26/H28), 6.46 (s, 2H, H11/H23), 6.22 (s, 2H,
H5/H17), 6.00 (s, 2H, H25/H27), 5.95 (d, J
34-33
= 8.3 Hz, 2H, H34/H50), 5.43 (dd, J
33-32
= 11.4 Hz, J
33-34
= 8.3 Hz, 2H,
H33/H49), 5.09 (dd, J
2-29’
= 12.5 Hz, J
2-29
= 4.8 Hz, 2H, H2/H14), 4.90 (dd, J
32-33
= 11.4 Hz, J
32-31
= 7.5 Hz, 2H, H32/H48),
4.65 (dd, J
8-36’
= 13.3 Hz, J
8-36
= 4.1 Hz, 2H, H8/H20), 3.86 (s, 6H, H10'/H22’), 3.86 (s, 6H, H12'/H24’), 3.57 (s, 6H,
H6'/H18’), 3.29 (s, 6H, H4'/H16’), 3.24 (dd, J
29-29’
= 14.2 Hz, J
29-2
= 4.8 Hz, 2H, H29/H45), 3.00 (dd, J
36-36’
= 17.3 Hz, J
36-8
=
4.1 Hz, 2H, H36/H52), 2.84 (dd, J
36’-36
= 17.3 Hz, J
36’-8
= 13.3 Hz, 2H, H36’/H52’), 2.38 (dd, J
29’-29
=14.2 Hz, J
29’-2
= 12.5
Hz, 2H, H29’/H45’).
13 C NMR (150 MHz, CDCl
3
, 10 mM, 298 K):

= 174.6 (C30/C46), 171.5 (C35/51), 157.3 (C6/C18), 156.6 (C4/C16), 155.9
(C10/C22), 154.8 (C12/C24), 140.0 (C37/C53), 137.3 (C41/C57), 128.8, 128.2, 128.1, 127.5, 127.1 (C37/C53, C38/C54,
C39/C55, C40/C56, C41/C57, C42/C58, C43/C59, C44/C60), 126.4 (C25/C27), 126.2 (C26/C28), 125.1 (C1/C13), 124.3
(C9/C21), 121.4 (C3/C15, C7/C19), 97.7 (C5/C17), 94.4 (C11/C23), 60.3 (C32/C49), 58.1 (C33/C49), 55.9 (C4’/C16’), 55.8
(C6’/C18’), 55.7 (C10’/C22’), 55.3 (C12’/C24’), 41.8 (36/C52), 40.1 (C29/C45), 33.5 (C2/C14), 33.4 (C8/C20).
6
OH o m
OH a p O
1 H NMR (600 MHz, CDCl
3
, 10 mM, 298 K):

= 7.46 (d, J o-m
= 7.2 Hz, 2H, Ho), 7.41-7.38 (m, 3H, Hm and Hp), 5.26 (s,
1H, Ha).
1 / 6
2 / 6
3 / 6
4 / 6
5 / 6
Table S1 . Mandelic acid ( 6 ) complexation shifts (

=
 obs
–
 f
, ppm) in CSA/ 6 mixtures. mixture

R
(ppm)
Ha

S
(ppm)

R
(ppm)
Ho

S
(ppm)
-0.03
0
-0.10
-0.12
-0.10
-0.03
0.01
-0.15
-0.08
-0.11
0.02
0.06
-0.05
-0.01
-0.03
0.03
0.03
-0.03
-0.04
-0.01

Table S2.
Diamide 1 complexation shifts (

=
 obs
–
 f
, ppm) in the 1 /( RS )6 mixture.
H

(ppm)
NH2
H6eq
0.16
-0.05
H7eq
H7ax
-0.03
-0.05
H6ax -0.03
Table S3.
Diamide 2 complexation shifts (

=
 obs
–
 f
, ppm) in the 2 /( RS )6 mixture.
H 
(ppm)
NH2 0.15
H5c -0.02
Table S4. Diffusion coefficients (D, x 10 10 m 2 s -1 ) of molecular tweezers CSAs and 6 in CSA/( RS )6 mixtures in comparison with pure compounds .
Sample D
CSA
D
( R )-6
D
( S )-6
6
1
1 /( RS )6
-
9.9
8.7
8.6
11.7
-
8.6
-
11.7
-
8.6
- 2
2 /( RS )6 7.4 8.5 8.5
Table S5.
Resorcin[4]arene 3 amide protons complexation shifts (

=
 obs
–
 f
, ppm) in the
( R )6 / 3 and ( S )6 / 3 mixtures (600 MHz, CDCl
3
, 10 mM, 298 K).
proton

(ppm) ( R )6 / 3

(ppm) ( S )6 / 3
NH31/NH43 0.03 0.05
NH34/NH46 0.13 0.17
Table S6.
Resorcin[4]arene 4 amide protons complexation shifts (

=
 obs
–
 f
, ppm) in the
( R )6 / 4 and ( S )6 / 4 mixtures (600 MHz, CDCl
3
, 10 mM, 298 K).
proton

(ppm) ( R )6 / 4

(ppm) ( S )6 / 4
NH31/NH47
NH34/NH50
0.06
0.17
0.06
0.13
Table S7. Diffusion coefficients (D, x 10 10 m 2 s -1 ) of resorcin[4]arene CSAs and of 6 in CSA/( RS )6 mixtures in comparison with pure compounds and evaluation of the bound state molar fraction (x b
) of 6 .
Sample D
CSA
D
( R )-6
D
( S )-6 x b ( R )-6 x b ( S )-6
6
3
3 /( RS )6
4
4 /( RS )6
5
5 /( RS )6
-
4.7
4.7
4.6
4.5
4.4
4.4
11.7
-
6.6
-
6.3
-
6.8
11.7
-
6.5
-
6.5
-
6.7
-
-
0.73
-
0.76
-
0.67
-
-
0.74
-
0.73
-
0.68

Table S8.
Mandelic acid benzylic and ortho protons non-equivalences (|

R
-

S
|, Hz) and diffusion coefficients (D, x 10
H
2
O (μL)*
10 m 2 s
|

R
-

-1 ) measured on the ( RS )6 / 4 sample (1:1, 10 mM) after addition of H
2
O.
S
|
CH
|

R
-

S
| o
D
( R )-6
D
( S )-6
0 24.6 16.5 6.3 6.5
2
6
22.2
21.0
16.2
15.9
6.5
6.5
6.7
6.8
* The addition of 2 μL of H
2
O in the sample corresponds to a molar ratio 4 /H
2
O=1:16.
Table S9.
Resorcin[4]arene 5 amide protons complexation shifts (

=
 obs
–
 f
, ppm) in the ( RS )6 / 5 mixture (600 MHz,
CDCl
3
, 10 mM, 298 K).
H

(ppm)
NH31/NH49
NH38/NH56
0.05
0.11
H H
O
H
NH
7
6
1
O
NH 2
3
H
4
5t H H
5c
1
5c
H H
5t
4 H
3
2 HN
O
1
6
7
8
9
H
H
HN
H
2
O
H1
H7
H4
H4 H1
H6 eq
H6 ax
/H7 ax
H5 t
NH a
7.0
6.5
6.0
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
ppm
H5 t
7.4
7.2
7.0
6.8
6.6
6.4
6.2
6.0
5.8
5.6
5.4
5.2
ppm
NH b
Figure S1 . 1D ROESY spectra (600 MHz, CDCl
3
, 10 mM, 298 K, mixing time = 500 ms) corresponding to the inversion of the amide protons in ( R,R )1 (a) and in ( S,S )2 (b).

H28
39
38
40
34 HN
37
33
32
31 HN
O
6'
O
35
O
6
4'
30
5
4
O
36/36'
B
7
3
8
O
10
28
2
10'
29/29'
O
24
24'
9
1
12'
11
A'
12
O
41/41'
27
13
25
A
23
14
26
15
19
O
B'
16
17
16'
18
O
18'
21
22
O
20
48/48'
47
O
NH
44
45
43
NH 46
49
52
22'
O
42
3
50
51
H29’
H33
H32
NH31 g
44
40
43
42
41
37
39
38
34
HN
33
32
HN
31
10'
24'
4
12'
O
6'
4'
O
35
5
O
6
30
4
O
36/36'
B
7
3
8
O
10
28
2
29/29'
O
24
11
9
12
O
45/45' 46
O
1
A'
27
13
25
A
23
14
15
O
16
16'
26 B' 17
19
21
22
O
20
52/52'
18
18'
O
51
O
NH
48
49
47
NH 50
53
57
54
58
22'
56
55
59
60
H28
H33
H32
H29’
H28
H8
H33
H32
H40eq H40ax
NH34 f
H38
H33 H32
H8
H28
H25
H24’
H29
H4’
H29’
H2 e
H28
H25
H34
H24’
H4’
H29
H34
H28
H27
H10’ H6’
H36
H36’
H8 d
H28
H27
H36
H10’ H6’
H36’
H28
H28
H2 H8
H4’
H6’
H36’
H29’
H25 c H2 H8
H4’
H6’
H36’ H29’
H25
H36’
H29’
H31
H34 H2 H8
H28 b
H31
H25
H33 H32
H36’
H29’
H5
H5
H16’ H16’
H27
H6’ a
H27
H2 H8
Figure S2.
1D ROESY spectra (600 MHz, CDCl
3
, 10 mM, 298 K, mixing time = 500 ms) corresponding to the inversion of selected protons of 3 and 4
: H6’ (a), H28 (b), H25 (c), H8 (d), H2 (e), NH34 (f) and NH31 (g).

H25 d
NH31 c
46
45 44
O
36
37
O
35
42
O
33
O
32
41
43
34
H
N
38
10' 12'
6'
4'
N
H 31
5
O
6
4
39
O
30
B
O
40/40'
7
3
8
O
10
28
2
29/29'
O
O
24
1
11
A'
9
27
13
12
O
O
47/47'
48
14
25
21
26
15
19
20
O
B'
16
17
18
O
A
23
22
O
58/58'
O
57
24' 22'
5
H
N
16'
18'
N
H
49
56
50
O
61
59
51
O
O
55
54
O
64
52
53
62
60
63
H28
H28
H4’/
H18’
H2/H8
H29
H40
H40’
H41
H2
H32
H29
H40
H40’ H29’
H42
H43
H2/H8 b
H28 NH31
H25
H25
H29
H40
H40’
H29’
H28 a
NH31
Figure S3 . 2D ROESY traces (600 MHz, CDCl
3
, 10 mM, 298 K, mixing time = 500 ms) corresponding to the inversion of selected protons of 5 : H28 (a), H2/H8 (b), NH31 (c) and H25 (d).
H36 c
( R,R )3
3.15
3.10
3.05
3.00
2.95
2.90
H36’
H36’
2.80
2.75
2.70
2.65
b
( R )6 /( R,R )3
H36
H36’ a
( S )6 /( R,R )3
3.15
3.10
3.05
3.00
2.95
2.90
2.85
2.80
2.75
2.70
2.65
ppm ppm
H36 H36’
( S,S )4
3.15
3.10
3.05
3.00
2.95
H36
H36’
2.90
2.85
2.80
2.75
2.70
2.65
( R )6 /( S,S )4
H36
H36’
( S )6 /( S,S )4
3.15
3.10
3.05
3.00
2.95
2.90
2.85
2.80
2.75
2.70
2.65
ppm ppm a c b
Figure S4. 1 H NMR (600 MHz, CDCl
3
, 10 mM, 298 K) spectral regions of methylene bridge protons H36 and H36’ of resorcin[4]arene 3
(left) in (a) ( R,R )3 , (b) ( R )6 / 3 and (c) ( S )6 / 3 and of resorcin[4]arene 4 (right) in (a) ( S,S )4 , (b) ( R )6 / 4 and (c) ( S )6 / 4 .
3.15
3.10
3.05
3.00
2.95
2.90
2.85
2.80
2.75
2.70
2.65
ppm
3.15
3.10
3.05
3.00
2.95
2.90
2.85
2.75
2.70
2.65
ppm 2.80

Cbz
H
N
O
OH
+ HO
OH
BOP, Et
3
CH
2
Cl
2
N,
, 4h, r.t.
Cbz
H
N
O
O
( S , S )8
O
O
N
H
Cbz
H
2,
Pd/C,
EtOH/THF 1:1
H
2
N
O
O
( S , S )9
O
O
NH
2
( S )7
Scheme S1 . Synthesis of intermediates ( S , S )8 and ( S , S )9 .
ClOC
O O
COCl
H
N O
O O
O
H
N
O
O
O O
O O
O
O
DIPEA, ( S , S )9 , dry THF, reflux
O
O O O
ClOC
O O
COCl
O
O
N
H
O
O O
O
N
H
O
O
( S , S ) -5
Scheme S2 . Synthesis of ( S , S )5 .