Bioorganic & Medicinal Chemistry Letters 29 (2019) 59–61

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Bioorganic & Medicinal Chemistry Letters

journal homepage: www.elsevier.com/locate/bmcl

Towards lead compounds as anti-cancer agents via new phaeosphaeride T

A derivatives
Victoria V. Abzianidzea, , Sofya A. Zakharenkovab, Natalia I. Moiseevac, Petr P. Beltyukova,
⁎

Valeriy A. Polukeevd, Yaroslav A. Dubrovskiia,e, Victor A. Kuznetsova, Yuri G. Trishinb,

Jennifer E. Mejiaf, Alvin A. Holderf,
⁎

a
Research Institute of Hygiene, Occupational Pathology and Human Ecology, Federal Medical Biological Agency, p/o Kuz’molovsky, Saint Petersburg 188663, Russian
Federation
b
Saint Petersburg State University of Industrial Technologies and Design, Saint Petersburg 198095, Russian Federation
c
N.N. Blokchin National Medical Research Center of Oncology, Moscow 115478, Russian Federation
d
Institute of Experimental Medicine, Saint Petersburg 197376, Russian Federation
e
Saint Petersburg State University, Institute of Chemistry, Saint Petersburg 198504, Russian Federation
f
Department of Chemistry and Biochemistry, Old Dominion University, 4541 Hampton Boulevard, Norfolk, VA 23529, United States

ARTICLE INFO ABSTRACT

Keywords: New derivatives of phaeosphaeride A (PPA) were synthesized and characterized. Anti-tumor studies were carried
Natural products out on the U937, HCT-116, PC3, MCF-7, A549, К562, NCI-H929, Jurkat, THP-1, RPMI8228 tumor cell lines, and
Phaeosphaeride A on the HEF cell line. All the compounds synthesized were found to have better efficacy than PPA towards the
In vitro studies tumor cell lines mentioned. Compound 6 (IC50 = 0.59 ± 0.27 µM) was observed to be 11 times more active
Anti-cancer agents
than PPA (IC50 = 6.5 ± 0.30 µM) towards the NCI-H929 cell line, with a therapeutic index of 18. Compound 6
Etoposide
was determined to be over half and 16 times more active than etoposide towards the NCI-H929
HEF cell line
HCT-116 cell line (IC50 = 0.9 ± 0.05 µM) and A549 (IC50 = 100 ± 7.0 µM) cell lines, respectively.
Mercapto derivative
Therapeutic index

Cancer, specifically malignant tumors, is a leading cause of death
around the world. The treatment of cancer dominates health care
budgets internationally and is a growing socio-economic threat against
humanity. Although chemotherapy has improved since its initial de-
velopment, the disabling side effects and relative effectivity of these
modern drugs have much room for improvement. With the aid of the
continuous addition of data from decades of chemotherapy research
and inspiration from successes seen in such drugs as, irinotecan,1 a
semisynthetic derivative of camptothecin, and etoposide,2 a semisyn-
thetic derivative of podophyllotoxin, scientists have begun to target the
chemical modification of natural anti-tumor agents. Phaeosphaeride A
(PPA) belongs to this same group of natural compounds and has an-
ticancer properties. This natural product was first isolated from the
endophytic fungus, FA39 (Phaeosphaeria avenara) by Clardy and co-
workers in 2006 and was reported to inhibit STAT3/DNA binding with
an IC50 value of 0.61 mM.3
In this work, new C-6 PPA derivatives (compounds 1–6) were syn-
thesized and characterized. Cytotoxic studies were carried out on 10
cell lines, e.g., HCT-116, PC3, MCF-7, A549, К562, NCI-H929, Jurkat,
THP-1, RPMI8228, and HEF.
The work of our group with PPA began with the production of new
derivatives formed in reactions involving active sites of a nitrogen-
containing heterocycle and a hydroxyl group at the C-6 atom.4–6 The
chemical modification of the nitrogen-containing ring was found to
lower the cytotoxic activity of the new derivatives towards A549 lung
cancer cells, while among the C-6 derivatives, a compound with greater
activity than the original phaeosphaeride was revealed (atom num-
bering is that of Clardy et al.3).
The selective fluorination of bioactive molecules is a well-proven
strategy in the development of potent drugs with improved bioavail-
ability, increased pharmaceutical efficacy, and reduced toxicity.7,8
Therefore, the secondary hydroxy group at the C-6 carbon was chosen
for substitution with a fluorine atom. Diethylaminosulfur trifluoride
(DAST), a nucleophilic fluorinating reagent, was used to fluorinate the
PPA. The reaction was carried out at 0 °C in anhydrous DCM (Scheme
1). Clean configurational inversion was observed. The ROESY spectrum

⁎
Corresponding authors.
E-mail addresses: vvaavv@mail.ru (V.V. Abzianidze), aholder@odu.edu (A.A. Holder).

https://doi.org/10.1016/j.bmcl.2018.11.003
Received 6 September 2018; Received in revised form 2 November 2018; Accepted 6 November 2018
Available online 09 November 2018
0960-894X/ © 2018 Elsevier Ltd. All rights reserved.
V.V. Abzianidze et al. Bioorganic & Medicinal Chemistry Letters 29 (2019) 59–61

Scheme 1. The fluorination of PPA.

N
O O HS O
Cl(CH2)nCOCl X
N O N O N O
HO HO K2CO3 HO
OH O
O O O n=1 O
O O
PPA 2: n = 1
3: n = 4 ( )n 2,3 4-6
Cl S

X N 4: X = O
5: X = NH
6: X = S
Scheme 2. Synthesis of products 2–6.

Table 1 of CH2 protons at the ester group (δ 4.18 and 4.09, J = 16.4 Hz) shown
IC50 values for the respective compounds when studied on the adhesive cell in the spectrum of 4. Another spin system consisted of two protons of a
lines. terminal alkene (δ 5.06 and 5.02, J = 1.4 Hz). The low-field signals of
Compound Adhesive cell cultures, IC50 (μM) mercapto substituents gave new chemical shifts in the 1H NMR spectra.
The cytotoxic activity of phaeosphaeride A and its derivatives
HCT-116 PC3 MCF-7 A549 HEF (compounds 1–6) were evaluated on the HCT-116, PC3, MCF-7, A549,
К562, NCI-H929, Jurkat, THP-1, RPMI8228 tumor cell lines and the
PPA 44 ± 0.75 32 ± 0.7 20 ± 0.8 41 ± 2.6 19 ± 0.25
1 31 ± 1.4 12 ± 1.7 37 ± 1.2 42 ± 0.7 37 ± 0.8 HEF cell line.
2 2.4 ± 0.18 2.4 ± 0.04 17 ± 1.4 20 ± 0.34 9.1 ± 0.23 Etoposide, a well-known anti-tumor drug, is a semisynthetic deri-
3 11 ± 0.48 9.2 ± 0.18 5.4 ± 0.15 12 ± 0.35 11 ± 0.52 vative of podophyllotoxin. It is used in combination with other che-
4 4.9 ± 0.21 5.8 ± 0.53 3.9 ± 0.02 20 ± 0.48 9.3 ± 0.26 motherapeutic agents for the treatment of refractory testicular tumors
5 8.2 ± 0.27 6.4 ± 0.36 16 ± 0.37 19 ± 0.46 11 ± 0.15
6 4.7 ± 0.09 5.0 ± 0.16 6.3 ± 0.15 5.9 ± 1.1 11 ± 0.2
and as a first line treatment in patients with small cell lung cancer.13
Etoposide 22 ± 1.1 2.7 ± 0.05 9.6 ± 0.27 100 ± 7.0 > 100 Etoposide is also used to treat other malignancies, such as lymphoma,
non-lymphocytic leukemia, and glioblastoma multiforme. For these
reasons, etoposide was used as the control during in vitro studies.
of 1 showed a correlation between the methyl protons at δ = 1.07 ppm Evaluation of the effect of PPA derivatives on proliferative activity
(H-15) and the proton at δ = 4.90 ppm (H-6), confirming the inversion and the viability of eukaryotic cells was performed using the MTT test.
of configuration of the C-6 atom. Product 1, phaeosphaeride B (PPB) IC50 values were determined for adhesive and suspension tumor cell
was also fluorinated and obtained. lines, MCF-7 (breast cancer), PC-3 (prostate adenocarcinoma), HCT-116
The formation of the same product allowed us to assume that the (colon cancer), A549 (lung cancer), K562 (chronic myelogenous leu-
reaction, specifically fluorination, most likely proceeded via the SN1 kemia), THP-1 (acute monocytic leukemia), NCI-H929 and RPMI8226
mechanism. The yields of the products from the fluorination of PPA and (multiple myeloma) Jurkat (T-cell lymphoblastic leukemia). Human
PPB were 41% and 25%, respectively. embryonic fibroblasts were used as a control.
The structure of compound 2, previously proven promising in terms The results obtained proved that the fluoro derivative 1 was 2 times
of biological activity, was optimized by extending the hydrocarbon less toxic than PPA towards HEF cells and 3 times more toxic to PC3
chain containing chlorine.4 Acylation of PPA was carried out by treat- cancer cell (Table 1). There was no significant change on cytotoxic
ment with 5-chlorovaleroyl chloride and resulted in the formation of activity against other cell lines (Table 2).
product 3 (Scheme 2). Various sulfides, 2-mercaptobenzoxazole, 2- The length of the chloro hydrocarbon chain impacted the IC50 va-
mercaptobenzimidazole, and 2-mercaptobenzothiazole were used to lues in different cells. In several cases, the IC50 values for substance 3
substitute the chlorine atom on compound 2. These pharmacophores was higher than for 2 (for HCT-116 and PC3 cell lines) or was nearly
contributed to the emergence or intensification of various biological equal (in K562). In most cases, an increase in the length of the chloro
activities, such as antiviral, antimicrobial, anticonvulsant, antioxidant, hydrocarbon chain of the substituent was accompanied by a 30–70%
as well as anti-cancer properties.9–12 Substitution of chlorine (Scheme decrease in the IC50 value (MCF-7, A549, NCI-H929, Jurkat, THP-1, and
2) proceeded in the presence of potassium carbonate with acetonitrile RPMI8228 cell lines). Compound 2 was found to be approximately 20
as solvent resulted in the formation of products 4–6. HRMS, 1H, and 13C times more toxic towards the HCT-116 cancer cell line than PPA,
NMR spectra of 4–6 proved their structures and purity. Their 1H NMR though its therapeutic index was low (3.8). All of the compounds syn-
spectra were similar to the spectrum of compound 2, with two doublets thesized were more toxic to HEF when compared to etoposide.

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V.V. Abzianidze et al. Bioorganic & Medicinal Chemistry Letters 29 (2019) 59–61

Table 2
IC50 values for the respective compounds when studied on the suspension cell lines.
Compound Suspension cell cultures, IC50 (μM)

К562 NCI-H929 Jurkat THP-1 RPMI8228

PPA 20 ± 1.46 6.5 ± 0.30 10 ± 0.2 19 ± 0.25 9.3 ± 0.36

1 18 ± 0.54 3.9 ± 0.2 15 ± 0.47 19 ± 0.8 16 ± 0.6
2 7 ± 0.25 3.3 ± 0.16 9.6 ± 0.5 11 ± 0.5 9.7 ± 0.08
3 8.5 ± 0.1 2.5 ± 0.1 4.2 ± 0.2 4.6 ± 0.03 4.3 ± 0.15
4 4.5 ± 0.13 3 ± 0.2 2.6 ± 0.05 2.3 ± 0.05 5.1 ± 0.13
5 9.3 ± 0.3 2 ± 0.19 8.9 ± 0.12 5.3 ± 0.5 4.3 ± 0.18
6 4.3 ± 0.13 0.59 ± 0.27 2.3 ± 0.17 1.6 ± 0.05 2.0 ± 0.13
Etoposide 9.4 ± 0.28 0.9 ± 0.05 1.4 ± 0.06 1.0 ± 0.02 4.8 ± 0.23

A549 cancer cell lines compared to parent PPA (Figs. 1 and 2).
The IC50 value of 6 was 11 μM on normal cells (HEF) and for NCI-
H929 cells the therapeutic value was 18.6. Compound 6 appeared to be
1.5 times more active on NCI-H929 cells and many times more active on
A549 cells than the control substance, etoposide.
In summary, the introduction of a fluorine atom into the PPA mo-
lecule reduced the toxicity towards almost all of the lines studied. The
number of carbon atoms in the chlorine-containing chain impacted
cytotoxicity. This study led to the development of a lead compound, the
mercapto derivative 6. Further biological tests are currently being
conducted.

Acknowledgements
Fig. 1. IC50 values of PPA, 6, and etoposide for the NCI-H929 cell line. Values
are mean ± SD (n = 3). The authors would like to thank Dr. A. Berestetskiy (All-Russian
Institute of Plant Protection, Saint Petersburg, Russian Federation) who
generously provided the natural phaeosphaeride A.

Appendix A. Supplementary data

Supplementary data to this article can be found online at https://

doi.org/10.1016/j.bmcl.2018.11.003.

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