Synthesis and bioactivity of diketopiperazine PJ147 and its derivatives from Gliocladium sp. YUP08

Article abstract of DOI:10.1080/10286020.2014.916283

Concise total synthesis of diketopiperazine PJ147, obtained from mycelium of Gliocladium sp. YUP08, has been achieved in seven steps with 43.5% overall yield. Biological evaluation of PJ147 exhibited strong inhibiting activity against A375-S2, Hela, P388, A-549, HL-60, and BEL-7420 cell lines. Thus, eight derivatives of PJ147 with high water solubility were also synthesized to facilitate the in vivo bioassay of this kind of diketopiperazines. 2014

Full text of DOI:10.1080/10286020.2014.916283

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Synthesis and bioactivity of
diketopiperazine PJ147 and its
derivatives from Gliocladium sp. YUP08
Xue-Zheng Lia, Gang Chenbc, Hai-Feng Wangbc, Hui-Ming Huabc &
Yue-Hu Peibc
a Department of Pharmacy, Affiliated Hospital of Yanbian
University, Yanji 133000, China
b School of Traditional Chinese Materia Medica, Shenyang
Pharmaceutical University, Shenyang 110016, China
c Key Laboratory of Structure-Based Drug Design and Discovery
(Shenyang Pharmaceutical University), Ministry of Education,
Shenyang 110016, China
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Published online: 14 May 2014.
To cite this article: Xue-Zheng Li, Gang Chen, Hai-Feng Wang, Hui-Ming Hua & Yue-Hu Pei (2014)
Synthesis and bioactivity of diketopiperazine PJ147 and its derivatives from Gliocladium sp. YUP08,
Journal of Asian Natural Products Research, 16:7, 764-769, DOI: 10.1080/10286020.2014.916283
To link to this article: http://dx.doi.org/10.1080/10286020.2014.916283
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Journal of Asian Natural Products Research, 2014
Vol. 16, No. 7, 764–769, http://dx.doi.org/10.1080/10286020.2014.916283
Synthesis and bioactivity of diketopiperazine PJ147 and its derivatives
from Gliocladium sp. YUP08
Xue-Zheng Lia, Gang Chenbc*, Hai-Feng Wangbc, Hui-Ming Huabc and Yue-Hu Peibc
aDepartment of Pharmacy, Affiliated Hospital of Yanbian University, Yanji 133000, China; bSchool
of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016,
China; cKey Laboratory of Structure-Based Drug Design and Discovery (Shenyang Pharmaceutical
University), Ministry of Education, Shenyang 110016, China
(Received 16 October 2013; final version received 15 April 2014)
Concise total synthesis of diketopiperazine PJ147, obtained from mycelium of
Gliocladium sp. YUP08, has been achieved in seven steps with 43.5% overall yield.
Biological evaluation of PJ147 exhibited strong inhibiting activity against A375-S2,
Hela, P388, A-549, HL-60, and BEL-7420 cell lines. Thus, eight derivatives of PJ147
with high water solubility were also synthesized to facilitate the in vivo bioassay of this
kind of diketopiperazines.
Keywords: diketopiperazines; antitumor; water solubility
1. Introduction
2. Results and discussion
In our ongoing search for bioactive
metabolites from marine microorganisms,
the new diketopiperazine PJ147 (11) was
obtained from mycelium of Gliocladium
sp. YUP08 isolated from sea mud collected
in Rushan [1]. Biological evaluation of
PJ147 (11) exhibited strong inhibiting
activity against A375-S2, Hela, P388,
A-549, HL-60, and BEL-7420 cell lines
with IC50 values of 4.6, 3.4, 2.86, 1.37, 1.73,
and 3.2 mmol/l, respectively. Thus, the total
synthesis study of PJ147 has been achieved
in seven steps with 43.5% overall yield.
However, the low water solubility of PJ147
hampers the in vivo bioassay. Thus, seven
derivatives of PJ147 named PJ1471-1478
(12–18)withhighwatersolubilitywerealso
synthesized to facilitate the in vivo bioassay
of this kind of diketopiperazines. All the
seven derivatives showed strong inhibiting
activity against HL-60 cell lines by in vitro
bioassay. In this paper, we report the
synthesis and biological evaluation of all
the compounds synthesized.
Derivatives of PJ147 (12–18) were
synthesized according to Scheme 1. The
condensation of N-carbobenzyloxy-L-ala-
nine (1) with glycine methyl ester (2)
provided dipeptider (3). Hereby, a ring
closure condensation of 3 established the
construction of 3-methylpiperazine-2,5-
dione scaffold of 4 by hydrogenation
catalyzed by Pd/C. The bromination of the
hydroxyl group of 6 with phosphorus
tribromide gave 7, which then reacted with
4-hydroxy benzaldehyde (8) to give 5 and
reacted with 3,4-dihydroxyl benzaldehyde
(20) to give 21. Etherification of the
phenolic hydroxyl group of 21 gave 22,
which was then nucleophilically substi-
tuted with different amines, leading to the
formation of 23. Finally, the condensation
of benzaldehyde derivatives (5 and 23)
with (S)-1,4-diacetyl-3-methylpiperazine-
2,5-dione (9) gave the target compounds
11–18, respectively.
The cytotoxic activities of synthesized
compounds 12–18 with better water
*Corresponding author. Email: chengang1152001@163.com
q 2014 Taylor & Francis

Journal of Asian Natural Products Research
765

766
X.-Z. Li et al.
solubility were bioassayed against HL-60
cell lines and their IC50 values were 7.27,
3.84, 4.11, 4.95, 5.4, 6.28, 7.93 mmol/l,
respectively. Based on the cytotoxic
activities of these synthesized compounds
12–18, it seemed that the long chain
substituent at C-10 does not impact
ZCHO), 7.79 (2H, d, J ¼ 8.7 Hz, H-30,
50), 6.98 (2H, d, J ¼ 8.7 Hz, H-20, 60), 5.51
(1H, t, J ¼ 6.4 Hz, H-2), 5.10–5.13 (1H,
m, H-6), 4.62 (2H, d, J ¼ 6.5 Hz, H-1),
2.13–2.19 (4H, m, H-4, 5), 1.77 (3H, s,
CH3-10), 1.70 (3H, s, CH3-8), 1.62 (3H, s,
CH3-9); HR-ESI-MS: m/z 259.1699
[M þ H]þ (calcd for C17H23O2,
259.1698).
the cytotoxic activity of PJ147 (IC50
1.73 mmol/l).
¼
3. Experimental
3.3 (S,Z)-3-(4-(3,7-Dimethylocta-2,6-
dienyloxy)benzylidene)-6-
methylpiperazine-2,5-dione (11)
3.1 General experimental procedures
The melting point as recorded on a Fluke
51II apparatus (Fluke Co., American Fork,
UT, USA). Optical rotations were
measured on a P-E 241 MC (Perkin-
Elmer Co., Jena, Germany). IR spectra
were recorded on a Bruker IFS-55 infrared
spectrophotometer with KBr disks (Bruker
Co., Zurich, Switzerland). The NMR
spectral data were recorded on Bruker
A mixture of 4 (640 mg) and acetic
anhydride (10 ml) was refluxed for 8 h at
1308C. Acetic anhydride was removed
under reduced pressure to obtain brunneus
liquid. The residue was dissolved in DMF
(20 ml) and 5 (5.17 g) was added to the
solution. To the reaction solution, potass-
ium tert-butoxide in tert-butyl alcohol
(15 ml) was slowly added and then was
stirred overnight at room temperature.
Then a saturated NaCl solution (40 ml)
was added to the reaction mixture and a
white precipitate was formed. The mixture
was extracted with ether (40 ml), which
was then removed under vacuum later to
give a yellow liquid.
1
AV-600 (300 MHz for H) with TMS as
internal standard (Bruker Co.). HR-FAB-
MS data were measured on Micross Mass
Autospec-Ultima-TOF
spectrometer
(Bruker Co.). Silica gel GF254 for TLC
and silica gel (200–300 mesh) for column
chromatography were obtained from Qing-
dao Marine Chemical Company, Qingdao,
China.
To a solution of hydrazine hydrate in
DMF (20 ml) was added the yellow liquid,
and the mixture was allowed to react over
3 h. To the reaction mixture was added
saturated salt solution and then precipi-
tation was formed. The mixture was
extracted with ether (40 ml), and the
solvent was removed under vacuum. The
residue was resolved by dichloromethane
and filtered, and the filtrate was removed
under reduced pressure to obtain 11
3.2 (E)-4-(3,7-Dimethylocta-2,6-
dienyloxy)benzaldehyde (5)
To a solution of p-hydroxy benzaldehyde 8
and a.p. carbonate (3.45 g) in acetone, 7
(0.002 mol) was added at room tempera-
ture. The reaction mixture was vigorously
stirred at reflux for 5 h, and filtered. The
filtered solid was diluted with ethyl acetate
(50 ml) and dried over anhydrous Na2SO4
over night. The reaction mixture was
extracted by 10% potassium hydroxide
and saturated salt solution, respectively.
And the ethyl acetate was removed under
reduced pressure to obtain 5. Brunneus
liquid, m.p. 104–1068C; 1H NMR
(300 MHz, CDCl3) d: 8.63 (1H, s,
20
(1.20 g, 65.0%). White powder, ½aꢀD
256.6 (c ¼ 0.52, MeOH); 1H NMR
(300 MHz, DMSO-d6) dH: 9.78 (1H, s,
NH-1), 8.36 (1H, s, NH-4), 7.45 (2H, d,
J ¼ 8.6 Hz, H-10, 100), 6.96 (2H, d,
J ¼ 8.6 Hz, H-11, 110), 6.64 (1H, s, H-8),
5.42 (1H, t, J ¼ 6.2 Hz, H-14), 5.08 (1H, t,
J ¼ 6.3 Hz, H-18), 4.57 (2H, d,

Journal of Asian Natural Products Research
767
J ¼ 6.4 Hz, H-13), 4.11 (1H, q,
J ¼ 6.9 Hz, H-6), 2.04–2.08 (4H, m, H-
16, 17), 1.71 (3H, s, CH3-22), 1.63 (3H, s,
CH3-20), 1.57 (3H, s, CH3-21), 1.33 (3H,
d, J ¼ 6.9 Hz, CH3-7); HR-ESI-MS: m/z
J ¼ 6.9 Hz, H-6), 4.00 (2H, t, J ¼ 6.3 Hz,
H-25), 2.51 (2H, t, J ¼ 6.9 Hz, H-27), 2.44
(4H, q, J ¼ 7.2 Hz, H-28, 280), 2.03–2.06
(4H, m, H-18, 19), 1.77–1.79 (2H, m, H-
26), 1.68 (3H, s, CH3-24), 1.62 (3H, s,
CH3-23), 1.55 (3H, s, CH3-22), 1.32 (3H,
d, J ¼ 6.9 Hz, CH3-7), 0.92 (6H, t,
J ¼ 7.2 Hz, H-29, 290); HR-ESI-MS: m/z
369.2175
[M þ H]þ
(calcd
for
C22H29O3N2, 369.2178).
498.3329
[M þ H]þ
(calcd
for
3.4 PJ1471 (12)
C29H44O4N3, 498.3332).
By following the same procedure as
described for the preparation of 11, 12
was obtained from (E)-3-(3-(dimethyla-
mino)propoxy)-4-(3,7-dimethylocta-2,6-
dienyloxy)benzaldehyde (0.359 g), instead
of 5.
3.6 PJ1473 (14)
By following the same procedure as
described for the preparation of 12, 14
was obtained from (E)-4-(3,7-dimethy-
locta-2,6-dienyloxy)-3-(3-(pyrrolidin-1-
½aꢀD 260.3 (c ¼ 0.52, MeOH); 1H
20
NMR (300 MHz, DMSO-d6) dH: 9.78 (1H,
br s, NH-1), 8.35 (1H, s, NH-4), 7.09 (H, br
s, H-10), 7.03 (H, d, J ¼ 8.4 Hz, H-14),
6.95 (1H, d, J ¼ 8.4 Hz, H-13), 6.61 (1H,
s, H-8), 5.40 (1H, t, J ¼ 6.0 Hz, H-16),
5.06 (1H, t, J ¼ 6.0 Hz, H-20), 4.55 (2H, d,
J ¼ 6.0 Hz, H-15), 4.09 (1H, q,
J ¼ 6.9 Hz, H-6), 4.00 (2H, t, J ¼ 6.3 Hz,
H-25), 2.34 (2H, t, J ¼ 7.0 Hz, H-27), 2.11
(2H, s, H-28, 29), 2.03–2.06 (4H, m, H-18,
19), 1.81–1.83 (2H, m, H-26), 1.68 (3H, s,
CH3-24), 1.62 (3H, s, CH3-23), 1.55 (3H,
s, CH3-22), 1.32 (3H, d, J ¼ 6.9 Hz, CH3-
7); HR-ESI-MS: m/z 470.3021 [M þ H]þ
(calcd for C27H40O4N3, 470.3019).
yl)propoxy)benzaldehyde
(0.385 g)
20
(0.163 g, 32.8%). White powder, ½aꢀD
259.3 (c ¼ 0.50, MeOH); 1H NMR
(300 MHz, DMSO-d6) dH: 9.78 (1H, br s,
NH-1), 8.35 (1H, s, NH-4), 7.07 (1H, br s,
H-10), 7.04 (1H, d, J ¼ 8.4 Hz, H-14),
6.95 (1H, d, J ¼ 8.4 Hz, H-13), 6.62 (1H,
s, H-8), 5.40 (1H, t, J ¼ 6.0 Hz, H-16),
5.04 (1H, t, J ¼ 6.0 Hz, H-20), 4.55 (2H, d,
J ¼ 6.0 Hz, H-15), 4.10 (1H, q,
J ¼ 6.9 Hz, H-6), 4.00 (2H, t, J ¼ 6.3 Hz,
H-25), 2.51 (2H, t, J ¼ 7.2 Hz, H-27),
2.39–2.41 (4H, m, H-28, 280), 2.03–2.06
(4H, m, H-18, 19), 1.85–1.97 (2H, m, H-
26), 1.68 (3H, s, CH3-24), 1.64–1.66 (4H,
m, H-29, 290), 1.62 (3H, s, CH3-23), 1.55
(3H, s, CH3-22), 1.32 (3H, d, J ¼ 6.9 Hz,
CH3-7); HR-ESI-MS: m/z 496.3177
[M þ H]þ (calcd for C29H42O4N3,
496.3175).
3.5 PJ1472 (13)
By following the same procedure as
described for the preparation of 12, 13
was obtained from (E)-3-(3-(diethyla-
mino)propoxy)-4-(3,7-dimethylocta-2,6-
3.7 PJ1474 (15)
dienyloxy)benzaldehyde
(0.387 g)
20
(0.165 g, 33.1%). White powder, ½aꢀD
263.3 (c ¼ 0.53, MeOH); 1H NMR
(300 MHz, DMSO-d6) dH: 9.78 (1H, br s,
NH-1), 8.35 (1H, s, NH-4), 7.07 (1H, br s,
H-10), 7.03 (1H, d, J ¼ 8.4 Hz, H-14),
6.95 (1H, d, J ¼ 8.4 Hz, H-13), 6.61 (1H,
s, H-8), 5.40 (1H, t, J ¼ 6.0 Hz, H-16),
5.04 (1H, t, J ¼ 6.0 Hz, H-20), 4.54 (2H, d,
J ¼ 6.0 Hz, H-15), 4.09 (1H, q,
By following the same procedure as
described for the preparation of 12, 15
was obtained from (E)-4-(3,7-dimethy-
locta-2,6-dienyloxy)-3-(3-(piperidin-1-yl)
propoxy)benzaldehyde (0.399 g) (0.170 g,
20
33.5%). White powder, ½aꢀD 261.1
(c ¼ 0.61, MeOH); 1H NMR(300 MHz,
DMSO-d6) dH: 9.78 (1H, br s, NH-1), 8.35
(1H, s, NH-4), 7.06 (1H, br s, H-10), 7.03

768
X.-Z. Li et al.
(1H, d, J ¼ 8.4 Hz, H-14), 6.95 (1H, d,
J ¼ 8.4 Hz, H-13), 6.62 (1H, s, H-8), 5.40
(1H, t, J ¼ 6.0 Hz, H-16), 5.05 (1H, t,
J ¼ 6.0 Hz, H-20), 4.55 (2H, d, J ¼
6.0 Hz, H-15), 4.10 (1H, q, J ¼ 6.9 Hz,
H-6), 4.00 (2H, t, J ¼ 6.3 Hz, H-25), 2.36
(2H, t, J ¼ 7.2 Hz, H-27), 2.25–2.30 (4H,
m, H-28, 280), 2.02–2.07 (4H, m, H-
18,19), 1.81–1.83 (2H, m, H-26), 1.68
(3H, s, CH3-24), 1.62 (3H, s, CH3-23),
1.55 (3H, s, CH3-22), 1.44–1.47 (4H, m,
CH3-29, 290), 1.35–1.37 (2H, m, CH3-30),
1.33 (3H, d, J ¼ 6.9 Hz, CH3-7); HR-ESI-
MS: m/z 510.3330 [M þ H]þ (calcd for
C30H44O4N3, 510.3332).
was obtained from (E)-4-(3,7-dimethy-
locta-2,6-dienyloxy)-3-(3-morpholinopro-
poxy)benzaldehyde (0.401 g) (0.179 g,
20
35.0%). White powder, ½aꢀD 262.9
1
(c ¼ 0.60, MeOH); H NMR (300 MHz,
DMSO-d6) dH: 9.78 (1H, br s, NH-1), 8.26
(1H, s, NH-4), 7.17 (1H, br s, H-10), 7.07
(1H, d, J ¼ 8.4 Hz, H-14), 6.94 (1H, d,
J ¼ 8.4 Hz, H-13), 6.55 (1H, s, H-8), 5.40
(1H, t, J ¼ 6.0 Hz, H-16), 5.05 (1H, t,
J ¼ 6.0 Hz, H-20), 4.54 (2H, d, J ¼
6.0 Hz, H-15), 4.08 (1H, q, J ¼ 6.9 Hz,
H-6), 4.00 (2H, t, J ¼ 6.3 Hz, H-25),
3.55 (4H, t, J ¼ 4.5 Hz, H-29, 290), 2.40
(2H, t, J ¼ 6.9 Hz, H-27), 2.31–2.36
(4H, m, H-28, 280), 2.03–2.07 (4H, m,
H-18, 19), 1.84–1.86 (2H, m, H-26), 1.68
(3H, s, CH3-24), 1.62 (3H, s, CH3-23),
1.55 (3H, s, CH3-22), 1.30 (3H, d,
J ¼ 6.9 Hz, CH3-7); HR-ESI-MS: m/z
512.3120 [M þ H]þ (calcd for C29H42O5
N3, 512.3124).
3.8 PJ1475 (16)
By following the same procedure as
described for the preparation of 12, 16
was obtained from (E)-4-(3,7-dimethy-
locta-2,6-dienyloxy)-3-(3-(4-methylpiper-
idin-1-yl)propoxy)benzaldehyde (0.399 g)
20
(0.155 g, 30.1%). White powder, ½aꢀD
260.8 (c ¼ 0.49, MeOH); 1H NMR
(300 MHz, DMSO-d6) d: 9.78 (1H, br s,
NH-1), 8.35 (1H, s, NH-4), 7.06 (1H, br s,
H-10), 7.03 (1H, d, J ¼ 8.4 Hz, H-14),
6.95 (1H, d, J ¼ 8.4 Hz, H-13), 6.62 (1H,
s, H-8), 5.40 (1H, t, J ¼ 6.0 Hz, H-16),
5.05 (1H, t, J ¼ 6.0 Hz, H-20), 4.55 (2H, d,
J ¼ 6.0 Hz, H-15), 4.10 (1H, q,
J ¼ 6.9 Hz, H-6), 4.00 (2H, t, J ¼ 6.3 Hz,
H-25), 2.79 (2H, t, J ¼ 11.3 Hz, H-28,
280), 2.38 (2H, t, J ¼ 7.2 Hz, H-27), 2.03–
2.07 (4H, m, H-18, 19), 1.81–1.83 (2H, m,
H-28, 280), 1.82–1.84 (2H, m, H-26), 1.68
(3H, s, CH3-24), 1.62 (3H, s, CH3-23),
1.55 (3H, s, CH3-22), 1.52 (2H, br s, CH3-
29, 290), 1.32 (3H, d, J ¼ 6.9 Hz, CH3-7),
1.29–1.31 (1H, m, H-30), 1.08–1.11 (2H,
m, H-29, 290), 0.86 (3H, d, J ¼ 6.3 Hz,
CH3-31); HR-ESI-MS: m/z 524.3485
[M þ H]þ (calcd for C31H46O4N3,
524.3488).
3.10 PJ1477 (18)
By following the same procedure as
described for the preparation of 12, 18
was obtained from (E)-4-(3,7-dimethy-
locta-2,6-dienyloxy)-3-(3-(4-methylpiper-
azin-1-yl)propoxy)benzaldehyde (0.414 g)
20
(0.202 g, 38.6%). White powder, ½aꢀD
257.1 (c ¼ 0.71, MeOH); 1H NMR
(300 MHz, DMSO-d6) dH: 9.78 (1H, br
s, NH-1), 8.35 (1H, s, NH-4), 7.06 (1H,
br s, H-10), 7.03 (1H, d, J ¼ 8.4 Hz, H-
14), 6.95 (1H, d, J ¼ 8.4 Hz, H-13), 6.62
(1H, s, H-8), 5.40 (1H, t, J ¼ 6.0 Hz, H-
16), 5.05 (1H, t, J ¼ 6.0 Hz, H-20), 4.54
(2H, d, J ¼ 6.0 Hz, H-15), 4.10 (1H, q,
J ¼ 6.9 Hz, H-6), 4.00 (2H, t, J ¼ 6.3 Hz,
H-25), 2.38–2.42 (2H, m, H-27), 2.27–
2.33(8H, m, H-28, 280, 29, 290), 2.12 (2H,
s, CH3-30), 2.03–2.07 (4H, m, H-18, 19),
1.81–1.83 (2H, m, H-26), 1.68 (3H, s,
CH3-24), 1.62 (3H, s, CH3-23), 1.55 (3H,
s, CH3-22), 1.32 (3H, d, J ¼ 6.9 Hz,
CH3-7); HR-ESI-MS: m/z 525.3441
[M þ H]þ (calcd for C30H45O4N4,
525.3441).
3.9 PJ1476 (17)
By following the same procedure as
described for the preparation of 12, 17

Journal of Asian Natural Products Research
769
Innovative Research Team of the Ministry of
Education and Program for Liaoning Innova-
tive Research Team in University.
4. Bioactivity studies
The anti-tumor activities of 11–18 were
determined according to the procedure
described in our previous research [2].
References
[1] Y.F. Huang, L. Tian, H.M. Hua, and Y.H.
Pei, J. Asian Nat Prod. Res. 9, 197 (2007).
[2] G. Chen, N. Xu, Z.F. Li, Q.H. Zhang, H.H.
Wu, and Y.H. Pei, J. Asian Nat Prod. Res.
2, 453 (2010).
Acknowledgments
This paper was financially supported by
projects of Shenyang science and technology
plans F12-156-9-00 and by Program for