A novel hybrid drug between two potent anti-tubulin agents as a potential prolonged anticancer approach.
We report the design, synthesis and biological characterisation of a novel hybrid drug by conjugation of two tubulin inhibitors, a hemiasterlin derivative A (H-Mpa-Tle-Aha-OH), obtained by condensation of three non-natural amino acids, and cis-3,4',5-trimethoxy-3'aminostilbene (B). As we have previously demonstrated synergy between A and B, we used a monocarbonyl derivative of triethylene glycol as linker (L) to synthesise compounds A-L and A-L-B; via HPLC we analysed the release of its potential hydrolysis products A, A-L, B and B-L in physiological fluids: the hybrid A-L-B undergo hydrolysis in rat whole blood of the ester bond between A and L (half-life=118.2±9.5min) but not the carbamate bond between B and L; the hydrolysis product B-L was further hydrolyzed, but with a slower rate (half-life=288±12min). The compound A-L was the faster hydrolyzed conjugate (half-life=25.4±1.1min). The inhibitory activity of the compounds against SKOV3 ovarian cancer cell growth was analysed. The IC50 values were 7.48±1.27nM for A, 40.3±6.28nM for B, 738±38.5nM for A-L and 37.9±2.11nM for A-L-B. The anticancer effect of A-L-B was evidenced to be obtained via microtubule dynamics suppression. Finally, we stated the expression of the active efflux transporters P-gp (ABCB1) and MRP1 (ABCC1) in the human normal colon epithelial NCM460 cell line by reverse-transcription PCR. Via permeation studies across NCM460 monolayers we demonstrate the poor aptitude of A to interact with active efflux transporters (AET): indeed, the ratio between its permeability coefficients for the basolateral (B)→apical (A) and B→A transport was 1.5±0.1, near to the ratio of taltobulin (1.12±0.06), an hemiasterlin derivative able to elude AETs, and significantly different form the ratio of celiprolol (3.4±0.2), an AET substrate.
Department of Chemical and Pharmaceutical Sciences, University of Ferrara, 44121 Ferrara, Italy. Electronic address: paolo.marchetti@unife.it.
Paolo
P
Marchetti
Department of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, Italy. Electronic address: pvnbbr@unife.it.
Barbara
B
Pavan
Department of Chemical and Pharmaceutical Sciences, University of Ferrara, 44121 Ferrara, Italy. Electronic address: daniele.simoni@unife.it.
Daniele
D
Simoni
Department of Chemical and Pharmaceutical Sciences, University of Ferrara, 44121 Ferrara, Italy. Electronic address: riccardo.barucchello@unife.it.
Riccardo
R
Baruchello
Department of Chemical and Pharmaceutical Sciences, University of Ferrara, 44121 Ferrara, Italy. Electronic address: riccardo.rondanin@unife.it.
Riccardo
R
Rondanin
Department of Biomedical and Speciality Surgical Sciences, Surgery and Experimental Medicine, University of Ferrara, 44121 Ferrara, Italy. Electronic address: carlo.mischiati@unife.it.
Carlo
C
Mischiati
Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, 44121 Ferrara, Italy. Electronic address: giordana.feriotto@unife.it.
Giordana
G
Feriotto
Department of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, Italy. Electronic address: frl@unife.it.
Luca
L
Ferraro
School of Pharmacy, College of Medicine, National Taiwan University, Taipei 10050, Taiwan. Electronic address: lihchinghsu@gmail.com.
Lih-Ching
LC
Hsu
Lestoni Corp., Richmond, VA 23219-155, USA. Electronic address: richvain@gmail.com.
Ray M
RM
Lee
Department of Chemical and Pharmaceutical Sciences, University of Ferrara, 44121 Ferrara, Italy. Electronic address: dla@unife.it.
Alessandro
A
Dalpiaz
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000235:genetics; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D045744:Cell Line, Tumor; D004355:Drug Stability; D006801:Humans; D006868:Hydrolysis; D008099:Liver / Q000378:metabolism; D008297:Male; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology; D017208:Rats, Wistar; D012995:Solubility; D013267:Stilbenes / Q000737:chemistry / Q000494:pharmacology; D050257:Tubulin Modulators / Q000737:chemistry / Q000494:pharmacology
D018435:ATP Binding Cassette Transporter, Subfamily B; D000970:Antineoplastic Agents; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D009842:Oligopeptides; D013267:Stilbenes; D050257:Tubulin Modulators; C513055:ABCB1 protein, human; C529077:cis-3,4',5-trimethoxy-3'-aminostilbene; C467070:hemiasterlin; C421109:multidrug resistance-associated protein 1
Hemiasterlin; Hybrid drug; Hydrolysis; NCM460 cells; Permeation; Stilbene