Cyclic Dipeptide Based Cell-Penetrating Peptidomimetics For DNA Delivery

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Cyclic dipeptide based cell-penetrating peptidomimetics for

effective DNA delivery
Received 00th January 20xx,
Accepted 00th January 20xx Chilakapati Madhu, Chandrashekhar Voshavar, K. Rajasekhar, T. Govindaraju*
DOI: 10.1039/x0xx00000x
Published on 21 March 2017. Downloaded by Fudan University on 22/03/2017 07:07:23.
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Unnatural CDP-amino acid (kd) is used in the design and synthesis design was conceptualized by the incorporation of cyclic
of novel cell penetrating peptidomimetic (Kkd-5). The balanced dipeptide (CDP) blocks into a peptide containing charged
cationic nature and structural rigidity of Kkd-5 resulted in serum amino acids.
stability, non-toxicity to cells, interaction with membrane and CDPs, also known as 2,5-diketopiperazines, are the
DNA, all of which facilitated efficient cellular uptake and DNA simplest of all the cyclic peptides. They are ubiquitously found
delivery. in nature as secondary metabolites and occur either in simple
13
CDP form or are embedded in complex molecular systems.
Effective and secure delivery of drugs/DNA across the plasma
membrane into the cell or specific organelle within remains a
challenge in the area of biotechnology and biomedical
1
sciences. Most of the delivery systems fail to effectively
penetrate the plasma membrane and exhibit high cytotoxicity,
showing low proteolytic stability and poor transfection
efficiency. At this juncture, cell penetrating peptides (CPPs)
have been widely explored for effective cellular uptake and
2
cargo delivery. CPPs offer numerous advantages over other
synthetic delivery systems including low cytotoxicity, easy
3
synthesis, and relatively high delivery efficiency. Among all
CPPs, cationic CPPs (TAT peptide, penetratin, and R8) are
4
promising candidates for cargo delivery. Amphipathic
peptides are another class of well-documented CPPs that
exhibit cell penetration through electrostatic and hydrophobic
5
interactions with the cell membrane. Polyproline and proline-
rich peptides are a minor class of CPPs shown to cross the
plasma membrane due to their well-known polyproline type–II
6
(PP-II) rod-like helical structures. Although several of the
aforementioned CPPs and their combinations have been
7
reported, each of them suffers from various limitations. In
recent years, CPPs with unnatural amino acids or molecular
8
scaffolds have been developed; these include β-peptides,
9 10 11
ureido peptides, D-peptides, peptoids,
guanidinocarbonylpyrrole and cycloalanine containing
12
peptides. Despite these interesting developments, there is an
enormous demand for novel and effective CPPs. In this
context, we envisaged a novel cell-penetrating peptidomimetic
(CPP) with balanced cationic character and rigid structure. The
Fig. 1 (a) Structures of unnatural CDP-amino acid (kd) and peptidomimetics with
K/E at alternative positions (Kkd-5/Ekd-5). FITC-Kkd-5 and FITC-Ekd-5 are
Bioorganic Chemistry Laboratory, New Chemistry Unit, Jawaharlal Nehru Centre fluorescently labeled versions of Kkd-5 and Ekd-5, respectively. FITC-kd-4 is FITC
for Advanced Scientific Research, Jakkur P.O., Bengaluru, Karnataka 560064, conjugated tetramer of kd. (b) Schematic representation of Kkd-5 cellular uptake,
India. E-mail: tgraju@jncasr.ac.in DNA delivery and transfection. FITC = Fluorescein isothiocyanate, fluorescent
†Electronic supplementary information (ESI) available:, Experimental procedure, label.
characterization and additional data. See DOI: 10.1039/x0xx00000x
This journal is © The Royal Society of Chemistry 20xx J. Name., 2013, 00, 1-3 | 1

Organic & Biomolecular Chemistry Page 2 of 6
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CDPs exhibit a structurally rigid conformation, which imparts yields (75%) and ready to be used for solid phase peptide
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proteolytic stability and perhaps supports hydrogen bonding synthesis (SPPS) of oligomers and peptidomimetics. Next, the
DOI: 10.1039/C7OB00167C
14
interactions with target systems. Furthermore, self-assembly kd unit was incorporated into peptides containing K and E at
of CDPs through versatile molecular chains and layers has been alternate positions through manual solid phase peptide

15
exploited for numerous functional bionanostructures. synthesis (SPPS) to obtain Kkd-5 and Ekd-5, respectively. The
Herein, we report the design and synthesis of an unnatural purity and integrity of all the CDP-peptidomimetics were
CDP-amino acid (kd) and its incorporation into peptide chains proved by HPLC and HRMS (ESI).
containing lysine and glutamic acid at alternative positions to To evaluate the cellular uptake, a fluorescence tag
obtain novel peptidomimetics Kkd-5 and Ekd-5, respectively (fluorescein isothiocyanate: FITC) was covalently conjugated to
(Fig. 1). We evaluated the CDP-peptidomimetics, Kkd-5 and resin-bound Kkd-5 and Ekd-5 to prepare FITC-Kkd-5 and FITC-
Ekd-5 for their serum stability, cellular uptake, cell viability and Ekd-5, respectively (Scheme 1b). The synthesized CDP-
as a synthetic delivery system for DNA with subsequent peptidomimetics were evaluated for their proteolytic stability
transfection studies in different cell lines. in human blood serum (HBS). Kkd-5 or Ekd-5 (50 µM) were
CDP is a well-known by-product in the peptide synthesis. incubated with HBS at 37 °C and analyzed at various time
We envisaged that incorporating CDP into the peptide- points (6, 12, 24 and 48 h) by RP-HPLC (Fig. 2a). Kkd-5 showed
backbone will generate novel peptidomimetics with enhanced high stability against degradation by proteases, with more
bioavailability, viability and utility. We designed a CDP-based than 96% of the peptidomimetic remaining intact even after
unnatural amino acid kd, as shown in Fig. 1, which was 48 h in the HBS. In contrast, Ekd-5 exhibited relatively low
subsequently incorporated into peptidomimetics through proteolytic stability with 30% degradation observed after 48 h
standard peptide synthesis. The synthesis of kd was initiated (Fig. 2a). The exceptional serum stability of Kkd-5 is an
with the coupling of Fmoc-Asp(OtBu)-OH and H-Lys(Boc)-OMe important attribute compared to other CPPs that are highly
using HBTU/HOBt as coupling reagents. The Fmoc-protected susceptible to serum proteases. Although cationic CPPs are
dipeptide 1 (Scheme 1a) was subjected to intramolecular known to be viable DNA delivery agents, the high charge
7,16
cyclization by treating with 20% piperidine in dichloromethane density impairs their cell viability. Therefore, in cellulo
(DCM) to obtain Boc and OtBu-protected cyclic dipeptide 2. cytotoxicity of Kkd-5 and Ekd-5 was evaluated in HeLa cells.
The acid-sensitive Boc and OtBu groups were deprotected with The cell viability was assessed at 24 and 48 h time periods
trifluoroacetic acid (TFA), followed by Fmoc-protection of the upon treating the cells with varying concentrations (50-500
amine functionality using Fmoc-succinimide (Fmoc-OSu) to µM) of Kkd-5 and Ekd-5 in MTT assay. Interestingly, Kkd-5 was
obtain the Fmoc-protected CDP monomer (Fmoc-kd) in good found to be non-toxic to cells even at 500 µM concentration
after 48 h of treatment (Fig. 2b). On the other hand, Ekd-5
showed ~10, 18 and 23% toxicity corresponding to 100 µM,
250 µM and 500 µM concentrations after 48 h of treatment
(Fig. 2b). Thus, the observed excellent cell viability of Kkd-5
can be possibly attributed to balanced and low cationic charge
17
density.
Fig. 2 (a) Serum stability of peptidomimetics Kkd-5 and Ekd-5 at 50 µM in human

blood serum (HBS) for a period of 48 h. (b) Cytotoxicity evaluation of
peptidomimetics, Kkd-5 and Ekd-5 in HeLa cells at 48 h. (c) Cellular uptake studies
in HeLa cells. Quantitative cellular uptake of, Kkd-5 and Ekd-5 assessed by flow
Scheme 1 (a) Synthesis of unnatural Fmoc-protected CDP-amino acid (Fmoc-kd). cytometry study, relative mean fluorescence intensity (RMFI). (d) Confocal
(b) Solid phase synthesis of CDP-incorporated peptidomimetics (Kkd-5, Ekd-5, microscopy images for cellular translocation (HeLa cells) and localization of FITC-
FITC-Kkd-5 and FITC-Ekd-5). Fmoc = Fluorenylmethyloxycarbonyl. Kkd-5 and FITC-Ekd-5, scale bar = 50 µM.

2 | J. Name., 2012, 00, 1-3 This journal is © The Royal Society of Chemistry 20xx

Page 3 of 6
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Cellular uptake is one of the key requisites that any DOI: 10.1039/C7OB00167C
synthetic delivery system must possess for successful DNA
delivery and transfection. The cellular uptake of CDP-

peptidomimetics was evaluated in HeLa cells. FITC-Kkd-5 and
FITC-Ekd-5 were incubated for 4 h at varying concentrations
(0.5, 1, 5, 10 and 20 µM); for comparative study, commercially
available stearic acid-conjugated arginine(R)-octapeptide
(FITC-Stearyl-R8) was used as a positive control. The flow
cytometry analysis of cells showed moderate uptake of ~30%
at 5 µM and interestingly, cellular uptake increased
significantly to 70% and 97% corresponding to 10 and 20 µM
of FITC-Kkd-5, respectively (Fig. 2c and S3). FITC-Ekd-5 did not
enter the cells at low concentration (5 µM) while at higher (10
µM and 20 µM) concentrations exhibited poor cellular uptake
(Fig. 2c and S3). On the other hand, the positive control (FITC-
Stearyl-R8) showed marginally higher cellular uptake
compared to Kkd-5 (Fig. S2 and S3). However, previous reports
Fig. 3 (a) NMR spectra of Kkd recorded upon addition of increasing concentration
revealed that Stearyl-R8 is toxic to cells at a concentration as DOPC [ 0 (i), 1 (ii), 2.5 (iii), 5 (iv), 7.5 (v), 10 (vi), 15 (vii) and 20 mM (viii)]. Inset:
18
low as 3.9 µM. Therefore, non-toxic nature and high serum Molecular level interaction between Kkd and the phosphate groups of DOPC. (b)
stability of Kkd-5 together make it a superior CPP. The confocal Agarose gel electrophoresis of Kkd-5 and Ekd-5 associated with pDNA (pCMV-β-
microscopy images clearly showed efficient cellular uptake and gal) in gel retardation assay. In vitro transfection efficiencies of pDNA (p-CMV-
SPORT-β-gal) delivered with Ekd-5, Kkd-5 and Stearyl-R8 in (c) HeLa cells and (d)
accumulation of Kkd-5 in the cytoplasm while Ekd-5 failed to HEK293 cells, measured through β-galactosidase activity and plotted against
penetrate the plasma membrane (Fig. 2d). The cationic nature varying pDNA:CPP ratios (1:1, 1:2 and 1:4). The transfection values shown are the
of Kkd-5, which is due to five lysine units, enabled its attractive average of triplicate experiments performed with 4-6 data points.
interaction with the negatively charged plasma membrane;
indicated that Kkd-5 interacts with the lipid membrane and
this is responsible for its high cellular uptake. Herein, we note
alters its curvature, whereas Ekd-5 exhibits negligible
that similar synthetic protocols is used for fluorescent labeling
interaction (Fig. S7). Although, the phase-transition or type of
(FITC-Kkd-5) which can be adapted for covalently conjugating
curvature was not deduced upon addition of Kkd-5 to the
various cell impermeable drugs to Kkd-5 and efficiently deliver
model membrane, an alteration in the lipid bilayer of model
them into cells for therapeutic applications.
membrane was observed, as revealed by the minor change in
Next, we embarked on probing the role of kd units towards
the Bragg peaks intensity and peaks overlap in the
observed cellular uptake of Kkd-5. A tetramer of kd-labeled
diffractograms (compare intensities of Kkd-5 alone versus Kkd-
with FITC (FITC-kd-4) was synthesized, and its cellular uptake
5 + model membrane, Fig. S7). Together, NMR and SAXS
was assessed in HeLa cells (Fig. 1a, Scheme 2S). Interestingly,
studies showed that cellular uptake of Kkd-5 is not only driven
FITC-kd-4 (50 µM) showed cellular uptake, which indicated a
by electrostatic interactions between cationic K units and the
possible complementary role of kd units apart from cationic K
anionic membrane but also through specific hydrogen bonding
units in enhancing cellular uptake of the cationic Kkd-5 (Fig.
interaction between kd units and head group of lipids of the
S5). To unravel the molecular level interactions of kd units
plasma membrane. In literature, the cellular uptake of CPPs
with the lipid membrane that contribute to efficient cellular
has been attributed to various mechanisms based on their
uptake we performed NMR studies by titrating 1,2-dioleoyl-sn-
structural and chemical composition. Careful inspection of
glycero-3-phosphocholine (DOPC, lipid component of the
fluorescence images of FITC-Kkd-5 treated cells showed
plasma membrane), against Kkd (Scheme 1S), the repeating
compartmentalized fluorescence in the cytoplasm remarkably,
building block of Kkd-5. The NMR spectrum of Kkd (10 mM) in
such a cellular uptake process is generally observed in the case
DMSO-d6 showed chemical shift signals for amide protons (- 6c
of endocytosis (Fig. S6). To confirm the endocytosis-driven
NH) of kd at 7.78 and 8.18 ppm and amine protons (-NH2) of K
cellular uptake we performed an additional experiment where
at 7.70 ppm (Fig. 3a). These chemical shift signals of Kkd were
PC12 cells were treated with FITC-Kkd-5 (40 µM) at two
downfield shifted upon titrating with increasing concentration
different incubation temperatures of 4 °C and 37 °C. The cells
of DOPC (1, 2.5, 5, 7.5, 10, 15 mM) (Fig. 3a). This downfield
incubated at 37 °C showed efficient cellular uptake and
shifting of proton signals reveals that amide protons of kd and
localization of FITC-Kkd-5 in the cytoplasm, whereas at 4 °C,
amine protons of K participate in hydrogen bonding
negligible uptake was observed. This data suggested that Kkd-
interaction with DOPC, as shown in Fig. 3a. To gain further
5 enters cells through an energy-dependent endocytosis
insight into the interaction of Kkd-5 and Ekd-5 with the plasma 6c
pathway (Fig. S6).
membrane, we performed small angle X-ray scattering (SAXS)
To assess the interaction of CDP-peptidomimetics with
studies with model membrane structure (ESI). The Bragg peaks
DNA, we performed gel retardation assay by treating fixed
in the SAXS diffractograms indicate constructive interference
concentration of pDNA (pCMV-SPORT-β-gal) with variable
of X-rays diffracted by lipid bilayer arrangement. SAXS profiles

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concentrations of Kkd-5 or Ekd-5 (1:0.5, 1:1, 1:2, 1:4 and 1:8). through specific hydrogen bonding and View electrostatic
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The gel retardation data showed that Kkd-5 complexes with interaction, that eventually enhanced the
DOI:cellular uptake and
10.1039/C7OB00167C
pDNA in a concentration-dependent manner, as shown in Fig. transfection efficiency of Kkd-5. Taken together, our results
3b. At 1:0.5 and 1:1 ratios, Kkd-5 displayed more than 50% suggest that CDP-peptidomimetic Kkd-5 with optimized

binding, which was further enhanced to 70% and 80% for 1:2 cationic charge, rigidity, and selective hydrogen bonding
and 1:4 ratios, respectively (Fig. S8). Moreover, at 1:8 ratio the interaction is an attractive strategy to further develop specific
pDNA band completely disappeared (pDNA gain net positive CPPs for delivering small molecule drugs, proteins and genes
charge), which signifies the strong binding of Kkd-5 with pDNA. for various biomedical applications.
On the other hand, Ekd-5 did not show any significant We thank Prof. C. N. Rao for constant support, IYBA and
interaction with pDNA at any of the concentration ratios (Fig. special nanotech grants from DBT and DST, Govt. of India, Dr.
3b). Moreover, ethidium bromide (EtBr) displacement assay Vemula, inSTEM for HeLa cells, Prof. Ranga, JNCASR for flow
using calf thymus DNA (ctDNA) showed efficient interaction cytometry, Dr. Mohan Rao and Dr. Sankaranarayanan, CCMB
and condensation of DNA by Kkd-5 (Fig. S9). The role of CDP for SAXS and Dr. S. Marepally, CSCR for transfection studies.
(kd) in Kkd-5 towards its complexation with pDNA was
investigated by NMR study, wherein increasing concentrations
(1, 2, 4, 8 and 10 mM) of adenosine monophosphate (AMP) References

was titrated against a fixed concentration of Kkd (10 mM). The 1 (a) D. Luo and W. M. Saltzman, Nat. Biotech., 2000, 18, 33-
chemical shift signals of amide protons (-NH) in kd exhibited a 37; (b) R. T. Chacko, J. Ventura, J. Zhuang and S.
downfield shift while the amine protons (-NH2) of K initially Thayumanavan, Adv. Drug Deliv. Rev., 2012, 64, 836-851.
2 H. Margus, K. Padari and M. Pooga, Mol. Ther., 2012, 20,
downfield shifted and finally disappeared at 4 mM of AMP (Fig.
525-533.
S10). These changes in the chemical shifts established the 3 S. Zhang, Y. Xu, B. Wang, W. Qiao, D. Liu and Z. Li, J. Control.
interaction of Kkd with AMP through hydrogen bonding. Thus, Release, 2004, 100, 165-180.
both electrostatic and hydrogen bonding interactions of Kkd in 4 (a) A. D. Frankel and C. O. Pabo, Cell, 1988, 55, 1189-1193;
Kkd-5 contribute towards the complexation of pDNA and its (b) N. Schmidt, A. Mishra, G. H. Lai and G. C. Wong, FEBS
Lett., 2010, 584, 1806-1813; (c) N. Nischan, H. D. Herce, F.
delivery across the plasma membrane into the cells.
Natale, N. Bohlke, N. Budisa, M. C. Cardoso and C. P. R.
Subsequently, we studied delivery and in cellulo transfection Hackenberger, Angew. Chem. Int. Ed., 2015, 54, 1950–1953;
(pDNA) ability of Kkd-5 and Ekd-5. A reporter gene (pDNA) (d) Z. Qian, A. Martyna, R. L. Hard, J. Wang, G. Appiah-Kubi,
encoding p-CMV-SPORT-β-gal for β-galactosidase (β-Gal) C. Coss, M. A. Phelps, J. S. Rossman, D. Pei, Biochemistry,
enzyme was used to evaluate and quantify the transfection 2016, 55, 2601-2612.
5 (a) J. Fernandez-Carneado, M. J. Kogan, S. Pujals and E.
efficiency of Kkd-5, Ekd-5 and Stearyl-R8 in HeLa and HEK293
Giralt, Biopolymers, 2004, 76, 196-203; (b) J. J. Welch, R. J.
cell lines. Kkd-5 exhibited a significant increase in the Swanekamp, C. King, D. A. Dean and B. L. Nilsson, ACS Med.
transfection efficacy as a function of its concentration; a 2-fold Chem. Lett., 2016, 7, 584-589.
enhancement in transfection efficacy was observed with 4 6 (a) K. Sadler, K. D. Eom, J.-L. Yang, Y. Dimitrova and J. P. Tam,
times increase in Kkd-5 concentration (Kkd-5:p-CMV-SPORT-β- Biochemistry, 2002, 41, 14150-14157; (b) J. Fernandez-
Carneado, M. J. Kogan, S. Castel and E. Giralt, Angew. Chem.
gal, 1:4). Remarkably, the transfection efficiency of Kkd-5 was
Int. Ed., 2004, 43, 1811-1814; (c) J. Farrera-Sinfreu, E. Giralt,
found to be ~50% higher compared to the positive control, S. Castel, F. Albericio and M. Royo, J. Am. Chem. Soc., 2005,
Stearyl-R8 in HeLa cells (Fig. 3c). Similar transfection profiles 127, 9459-9468; (d) Y. A. Fillon, J. P. Anderson and J.
were observed in HEK293 cells, where Kkd-5 exhibited ~48% Chmielewski, J. Am. Chem. Soc., 2005, 127, 11798-11803; (e)
higher transfection efficiency over Stearyl-R8 (Fig. 3d). As M. Nanda and K. N. Ganesh, J. Org. Chem., 2012, 77, 4131-
4135; (f) Y. A. Nagel, P. S. Raschle and H. Wennemers,
expected, Ekd-5 failed to show any transfection (Fig. 3c and
Angew. Chem. Int. Ed., 2017, 56, 122-126.
3d). The remarkable transfection efficiency of DNA (p-CMV- 7 D. M. Copolovici, K. Langel, E. Eriste and U. Langel, ACS Nano,
SPORT-β-gal) by Kkd-5 can be attributed to its unique design 2014, 8, 1972-1994.
comprising a rigid CDP backbone and cationic lysine units at 8 (a) N. Umezawa, M. A. Gelman, M. C. Haigis, R. T. Raines and
alternative positions. It is worthwhile to mention that efficient S. H. Gellman, J. Am. Chem. Soc., 2002, 124, 368-369; (b) M.
Rueping, Y. Mahajan, M. Sauer and D. Seebach,
cellular uptake of Stearyl-R8 failed to translate into matching
ChemBioChem, 2002, 3, 257-259.
transfection efficiency as compared to Kkd-5 because the high 9 C. Douat, C. Aisenbrey, S. Antunes, M. Decossas, O. Lambert,
positive charge density of Stearyl-R8 plays a key role in the B. Bechinger, A. Kichler and G. Guichard, Angew. Chem. Int.
cellular uptake while concurrently imparting high toxicity to Ed., 2015, 54, 11133-11137.
the cells. 10 J. Zhou, X. Du, J. Li, N. Yamagata and B. Xu, J. Am. Chem. Soc.,
2015, 137, 10040-10043.
In summary, we demonstrated an efficient strategy to
11 P. A. Wender, D. J. Mitchell, K. Pattabiraman, E. T. Pelkey, L.
prepare cell-penetrating peptidomimetic (Kkd-5) using CDP. Steinman and J. B. Rothbard, Proc. Natl. Acad. Sci. USA, 2000,
The presence of CDP-unnatural amino acid (kd) in Kkd-5 97, 13003-13008.
imparted high serum stability, bioavailability and viability 12 (a) M. Li, S. Schlesiger, S. K. Knauer and C. Schmuck, Angew.
properties. Kkd-5 showed efficient cellular uptake and Chem. Int. Ed., 2015, 54, 2941-2944; (b) H. Wu, G. Mousseau,
S. Mediouni, S. T. Valente and T. Kodadek, Angew. Chem. Int.
transfection efficacy of pDNA (p-CMV-SPORT-β-gal) in HeLa
Ed., 2016, 55, 12637-12642.
and HEK293 cells. NMR and SAXS analysis showed the selective 13 A. D. Borthwick, Chem. Rev., 2012, 112, 3641-3716.
molecular level interaction of kd and K units with DOPC
4 | J. Name., 2012, 00, 1-3 This journal is © The Royal Society of Chemistry 20xx

Page 5 of 6
Journal Name COMMUNICATION
14 (a) R. B. Corey, J. Am. Chem. Soc., 1938, 60, 1598-1604; (b) J.

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Bernard and H. Wennemers, Org. Lett., 2007, 9, 4283-4286. DOI: 10.1039/C7OB00167C
15 (a) J. C. MacDonald and G. M. Whitesides, Chem. Rev., 1994,
94, 2383-2420; (b) T. Govindaraju and M. B. Avinash,
Nanoscale, 2012, 4, 6102-6117; (c) H. Geng, L. Ye, A. Zhang,

J. Li and Z. Feng, Langmuir, 2016, 32, 4586-4594; (d) S.
Manchineella and T. Govindaraju, ChemplusChem, 2016,
DOI: 10.1002/cplu.201600450.
16 S. El-Andaloussi, P. Jarver, H. J. Johansson and U. Langel,
Biochem. J., 2007, 407, 285-292.
17 S. Futaki, T. Suzuki, W. Ohashi, T. Yagami, S. Tanaka, K. Ueda
and Y. Sugiura, J. Biol. Chem., 2001, 276, 5836-5840.
18 S. Futaki, W. Ohashi, T. Suzuki, M. Niwa, S. Tanaka, K. Ueda,
H. Harashima and Y. Sugiura, Bioconjugate Chem., 2001, 12,
1005-1011.


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Graphical Abstract

Cyclic dipeptide based cell-Penetrating peptidomimetics for effective DNA Delivery

Chilakapati Madhu, Chandrashekhar Voshavar, K. Rajasekhar, T. Govindaraju*

Cyclic dipeptide as an unnatural amino acid employed in the preparation of novel cell penetrating peptidomimetics and their
effective DNA delivery is demonstrated.


Cyclic Dipeptide Based Cell-Penetrating Peptidomimetics For DNA Delivery

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