Synthesis and biopharmaceutical studies of jltn as potential dasatinib prodrug

Article abstract of DOI:10.1248/cpb.c13-00248

Dasatinib was identified as a potent orally administered Src/Abl kinase inhibitor with excellent antiproliferative activity against Philadelphia chromosome-positive chronic myelogenous leukemia in chronic phase. The low bioavailability of Dasatinib may be due to both incomplete oral absorption and first-pass metabolism. A prodrug, JLTN, was synthesized to minimize the first-pass effect of Dasatinib and improve the oral bioavailability following oral administration via targeting intestinal peptide transporter and enhancing chemical stability. Biological evaluation data indicated that there was a 150%-fold increase in oral bioavailability of this prodrug compared to the parent drug Dasatinib in monkeys.

Full text of DOI:10.1248/cpb.c13-00248

August 2013
Note
Chem. Pharm. Bull. 61(8) 877–881 (2013)
877
Synthesis and Biopharmaceutical Studies of JLTN as Potential Dasatinib
Prodrug
,a
Fei Liu,a Li-Wei Lang,a Ji Jiang,b Hua-Jun Lu,c Jian-Min Wang,c and Shih-Chen Wang*
a Department of Nuclear Medicine, Peking Union Medical College & Chinese Academy of Medical Sciences,
b
Peking Union Medical College Hospital; Clinical Pharmacology Research Center, Peking Union Medical College &
Chinese Academy of Medical Sciences, Peking Union Medical College Hospital; Beijing 100730, China: and
c BL Pharmaceuticals; 2–3 Industrial Park, Renhe Town, Shunyi, Beijing 101300, China.
Received March 31, 2013; accepted May 9, 2013; advance publication released online May 16, 2013
Dasatinib was identified as a potent orally administered Src/Abl kinase inhibitor with excellent anti-
proliferative activity against Philadelphia chromosome-positive chronic myelogenous leukemia in chronic
phase. The low bioavailability of Dasatinib may be due to both incomplete oral absorption and first-pass me-
tabolism. A prodrug, JLTN, was synthesized to minimize the first-pass effect of Dasatinib and improve the
oral bioavailability following oral administration via targeting intestinal peptide transporter and enhancing
chemical stability. Biological evaluation data indicated that there was a 150%-fold increase in oral bioavail-
ability of this prodrug compared to the parent drug Dasatinib in monkeys.
Key words Dasatinib; prodrug; JLTN
Dasatinib, [N-(2-chloro-6-methylphenyl)-2-({6-[4-(2-hydrox- Fig. 2, 144 compounds) as potential prodrugs of Dasatinib
yethyl)piperazin-1-yl]-2-methylpyrimidin-4-yl}amino)-1,3-thia- were designed with the aim of improving aqueous solubility
zole-5-carboxamide, monohydrate; formula is C22H26ClN7O2S· and therapeutic efficacy, in terms of use of computer-aided
H2O; see Fig. 1] is a novel, small molecule, orally-bioavailable drug design methodologies (small molecule flexible docking,
multi-targeted kinase inhibitor, developed by Bristol-Myers virtual screening, three dimensions (3D) database search, lead
Squibb (U.S.A.) for the treatment of adults with chronic phase optimization, 3D-quantitative structure–activity relationship
or advanced phase (accelerated, myeloid blast or lymphoid (3D-QSAR), pharmacophore generation and peptide transport-
blast phase) chronic myeloid leukemia (CML) and resistance er). According to the principles of PepT prodrug technique,
to or intolerance of prior therapy (including Imatinib). It in- after getting into the digestive system, the compounds should
hibits breakpoint cluster region/Abelson, v-src sarcoma viral rapidly be converted into Dasatinib by various enzymes in the
oncogene homolog, proto-oncogene c-kit, ephrin a receptor, digestive tract. The compounds interact with intestinal protein
platelet-derived growth factor receptors, and other tyrosine transporters so that the compounds can be easily absorbed in
kinases. Dasatinib is also indicated for the treatment of adults an intestinal tract in comparison with the Dasatinib to gener-
with Philadelphia chromosome-positive acute lymphoblastic ate higher Dasatinib bioavailability.9) Overall, a series of 42
leukemia (Ph+ALL) and lymphoid blast CML and resistance or compounds was synthesized and evaluated. Compared with
intolerance of prior therapy.1–8)
the existing antitumor compounds, the compound (JLTN)
However, Dasatinib suffers from low oral bioavailability showed better in vitro anticancer activities against several can-
(ca. 14%), has large individual treatment variability, and more cer cell lines than that of Dasatinib. Also, pharmacodynamics
adverse events than Imatinib. It is now used as a second line studies have showed that the compound has notable curative
medication for the treatment of CML and Ph+ALL patients. effect and lower side effect.11,12) In an effort to improve the
The incomplete oral bioavailability of Dasatinib may be low oral bioavailability of Dasatinib, detailed synthesis route for
due to poor absorption from the gastrointestinal tract and/ its prodrug (JLTN) will be presented. The oral bioavailability
or extensive first-pass metabolism. In addition, the literature of the prodrug (JLTN) was evaluated after oral administration
suggests that the incomplete oral bioavailability of Dasatinib in monkeys and compared to the parent drug Dasatinib.
is due to a combination of incomplete absorption and high
first pass metabolism.8) Peptide transporters (PepT) show suf-
ficiently high-capacity low-affinity specificity and appear to be
attractive targets for increasing intestinal absorption of some
small molecules. Han and Amidon described this prodrug
strategy (by using enzymatically hydrolyzable bonds in prepa-
ration of PepT-targeted prodrugs) as “peptide transport associ-
ated prodrug therapy.” Good examples of prodrugs are valacy-
Fig. 1. Two-Dimensional Structure of Dasatinib
clovir (Valtrex®, GlaxoSmithKline, U.K.; l-valylester prodrug
of acyclovir) and valganciclovir (Valcyte®, Roche, Swiss Con-
federation; l-valylester prodrug of ganciclovir). Therefore, it is
necessary to increase the absorption rate of Dasatinib as well
to decrease the rate of its metabolism.9–12)
In this current study, a series of amino acid derivatives (see
The authors declare no conflict of interest.
Fig. 2. A Series of Dasatinib Amino Acid Derivatives
© 2013 The Pharmaceutical Society of Japan
*To whom correspondence should be addressed. e-mail: gen_scwang@qq.com

878
Vol. 61, No. 8
Reagents and conditions: (a) 4,6-Dichloro-2-methylpyrimidine (1.5eq.), Cs2CO3 (2.0eq.), DMF, 25°C, 24h, 83%; (b) K2CO3 (1.5eq.), 1-(bromomethyl)-4-methoxyben-
zene (1.2eq.), DMF, 25°C, 18h, 46%; (c) NaOH–H2O (2.0eq., 20%), THF–CH3OH–H2O (3:3:1), 25°C, 20h, 70%; (d) thionyl dichloride (6.0eq.), THF, 50°C, 5h, then
(2-chloro-6-methylphenyl)amine (3.0eq.), 50°C, 18h, 80%; (e) piperazine (10.0eq.), 1,4-dioxane, reflux, 24h, 95%; (f) 2-bromoethanol-d4 (0.7eq.), N,N-diethylethanamine
(1.4eq.), DMF, 60°C, 24h, 57%; (g) N-(tert-butoxycarbonyl)-l-leucine (1.2eq.), EDC·HCl (1.5eq.), HOBT (1.5eq.), CH2Cl2, 25°C, 20h, 65%; (h) trifluoroacetic acid
(0.2eq.), CH2Cl2, 30°C, 48h; (i) hydrogen chloride–propan-2-ol (5.0eq.), ethyl acetate, 25°C, 0.5h, 78% (two steps).
Chart 1. Synthesis of JLTN-d4 (8)
d4, N-(2-chloro-6-methylphenyl)-2-({6-[4-(2-hydroxyethyl)-
Chemistry In the present study, the synthetic routes to the piperazin-1-yl]-2-methylpyrimidin-4-yl}amino)-1,3-thia-
Dasatinib amino acid derivative are outlined in Chart 1.13,14)
zole-5-carboxamide-d4; JLTN-d4, 2-(4-{6-[(5-{[(2-chloro-6-
Results and Discussion
JLTN-d4 (8) was prepared from (2E)-N-(2-chloro-6- methylphenyl)amino]carbonyl}-1,3-thiazol-2-yl)amino]-2-
methylphenyl)-2-({6-[4-(2-hydroxyethyl)piperazin-1-yl]-2- methylpyrimidin-4-yl}piperazin-1-yl)ethyl l-leucinate trihy-
methylpyrimidin-4-yl}imino)-3-(4-methoxybenzyl)-2,3- drochloride-d4 (8) were synthesized at BL Pharmaceuticals.
dihydro-1,3-thiazole-5-carboxamide-d4 (6) and N-(tert-
Conclusions and Discussion In order to improve the
butoxycarbonyl)-l-leucine through three steps, including aqueous solubility and therapeutic efficacy of Dasatinib, the
condensation, deprotection and salification. Compound 6 was prodrug JLTN was synthesized and evaluated bioavailability
obtained from the ethyl 2-amino-1,3-thiazole-5-carboxylate by by oral administration to monkeys. Biological evaluation data
nucleophilic substitution, protection, hydrolysis, amidation and indicated that there was a 150%-fold increase in oral bioavail-
alkylation with ethyl 2-bromoethanol-d4.
Reference standards for high-performance liquid chroma-
ability of this prodrug compared to the parent drug Dasatinib.
In conclusion, JLTN could improve the oral bioavailability
tography (HPLC) and mass spectrometry, namely, Dasatinib- of Dasatinib in monkeys through PepT-mediated absorption

August 2013
879
and enhanced chemical stability. The prodrug strategy tar- hydro-1,3-thiazole-5-carboxylic Acid To
a
solution of
geted to intestinal PepT could offer a promising strategy to compound 2 (25.0g) in CH3OH–tetrahydrofuran (THF)–H2O
improve oral bioavailability of poorly absorbed Dasatinib. (3:3:1, 350mL) was added dropwise a 5n aq. NaOH solution
Preliminary in vivo studies indicated that this prodrug has (25mL) at 5°C. The solution was warmed to room tempera-
significantly improved oral bioavailability (rat) than of the ture, stirred for 18h. The mixture was concentrated in vacuo,
reference drug,8) but additional pharmacokinetic studies of and the residue was washed with water, and dried in vacuo
JLTN for preclinical studies need to get confirmed by further to obtain compound 3 (16.3g, 69.8%) as a white solid. HPLC
investigations.
Purity: 91.2%. ESI-MS m/z (M+H+) Calcd 391.1, Obsd 391.4.
1H-NMR (600MHz, DMSO-d6) δ ppm 2.49 (3H, s), 3.66 (3H,
s), 5.22 (2H, s), 6.80–6.82 (3H, m), 7.31–7.32 (2H, m), 7.82
Experimental
General All reagents were purchased from commercial (1H, s). 13C-NMR (150MHz, DMSO-d6) δ ppm 25.18, 49.76,
manufacturers unless otherwise indicated. All chemicals used 55.00, 110.54, 114.96, 128.43, 129.77, 158.12, 158.86, 162.70,
were reagent grade or better.
164.06, 166.17.
Proton NMR spectra were recorded on a Bruker DRX-600
Synthesis of Compound 4, (2E)-N-(2-Chloro-6-
spectrometer using CDCl3 or dimethyl sulfoxide-d6 (DMSO- methylphenyl)-2-[(6-chloro-2-methylpyrimidin-4-yl)-
d6) as solvent at ambient temperature and tetramethylsilane imino]-3-(4-methoxybenzyl)-2,3-dihydro-1,3-thiazole-5-
(TMS) as an internal standard. All reactions were monitored carboxamide To a solution of compound 3 (15.0g, 1.0eq.)
by Merck Millipore classical Silica gel 60F254 TLC plates. All in dry THF (230mL) was added dropwise thionyl chloride
compounds were of >90% purity by analytical HPLC analy- (6.0eq.). The reaction mixture was heated to 45°C for about
ses.
All procedures used for the animal studies were approved peated evaporation with dry THF in vacuo. The crude acyl
by the BL Pharmaceuticals Animal Care and Use Committee.
chloride was dissolved in dry THF (240mL) and added drop-
5h. Then, the excess of thionyl chloride was removed by re-
Syntheses. Synthesis of Compound 1, Ethyl 2-[(6-Chloro- wise a solution of the 2-chloro-6-methylaniline (3.0eq.) in dry
2-methylpyrimidin-4-yl)amino]-1,3-thiazole-5-carboxylate THF (10mL). Then, the reaction mixture was stirred at 45°C
To an ice-cooled solution of ethyl 2-amino-1,3-thiazole-5-car- for about 18h. The reaction was monitored using TLC with
boxylate (35.0g, 1.0eq.), Cs2CO3 in N,N-dimethylformamide petroleum ether–ethyl acetate (5:1) as eluent. The solution
(DMF) (2.0eq., 250mL) was added dropwise a solution of was concentrated. The residue was suspended in 1n aq. hydro-
4,6-dichloro-2-methylpyrimidine in DMF (1.5eq., 70mL). gen chloride solution (150mL), washed with water. The resi-
The solution was warmed to room temperature, stirred for due was suspended in benzene–1,4-dioxane (6:1, 225mL) and
24h. The reaction was monitored using TLC with petroleum filtered. The residue was triturated with benzene. The solid
ether–ethyl acetate (2:1) as eluent. The mixture was poured was filtered and dried in vacuo to obtain compound 4 (15.8g,
onto crushed ice (1000g), vigorously stirred for 10min and 80.0%) as a tan solid. HPLC Purity: 97.9%. ESI-MS m/z (M+
collected by filtration. The residue was washed with water, H+) Calcd 514.1, Obsd 514.4. 1H-NMR (600MHz, DMSO-
ethyl acetate, and dried in vacuo to obtain compound 1 (50.7g, d6) δ ppm 2.20 (3H, s), 2.57 (3H, s), 3.73 (3H, s), 5.37 (2H,
83.5%) as a yellow solid. HPLC Purity: 96.2%. Electrospray s), 6.94–7.42 (7H, m), 8.38 (1H, s), 10.03 (1H, s). 13C-NMR
ionization mass spectrometry (ESI-MS) m/z (M+H+) Calcd (150MHz, DMSO-d6) δ ppm 18.25, 25.11, 50.32, 55.08, 111.14,
299.0, Obsd 299.3. 1H-NMR (600MHz, DMSO-d6) δ ppm 114.20, 118.34, 127.05, 127.90, 128.35, 129.10, 129.98, 131.11,
1.29 (3H, t, J=6.0Hz), 2.59 (3H, s), 4.28 (2H, q, J=6.0Hz), 132.14, 132.94, 138.59, 158.61, 158.70, 159.11, 161.69, 163.74,
6.94 (1H, s), 8.13 (1H, s), 12.36 (1H, s). 13C NMR (150MHz, 166.30.
DMSO-d6) δ ppm 14.23, 25.25, 60.74, 103.61, 121.59, 145.46,
157.39, 158.60, 161.49, 162.36, 167.40.
Synthesis of Compound 5, (2E)-N-(2-Chloro-6-
methylphenyl)-3-(4-methoxybenzyl)-2-[(2-methyl-6-piper-
Synthesis of Compound 2, Ethyl (2E)-2-[(6-Chloro- azin-1-ylpyrimidin-4-yl)imino]-2,3-dihydro-1,3-thiazole-
2-methylpyrimidin-4-yl)imino]-3-(4-methoxybenzyl)-2,3- 5-carboxamide To a solution of compound 4 (11.0g, 1.0eq.),
dihydro-1,3-thiazole-5-carboxylate To an ice-cooled solu- piperazine (10.0eq.) in 1,4-dioxane (110mL) was refluxed for
tion of compound 1 (48.3g, 1.0eq.), K2CO3 in DMF (1.5eq., 24h. The mixture was concentrated under vacuum, and the
480mL) was added dropwise 1-(bromomethyl)-4-methoxyben- solid was triturated successively with water and vigorously
zene (1.5eq.). The solution was warmed to room temperature, stirred for 10min. The precipitate was collected by filtration
stirred for 18h. The mixture was poured onto crushed ice and purified by column chromatography (CH2Cl2–CH3OH=
(500g), vigorously stirred for 10min and collected by filtra- 10:1) to give compound 5 (11.5g, 95.3%) as a light yellow
tion. The residue was washed with water, petroleum ether– solid. HPLC Purity: 93.3%. ESI-MS m/z (M+H+) Calcd 564.2,
1
ethyl acetate (1:2), and dried in vacuo to obtain compound 2 Obsd 564.4. H-NMR (600MHz, CDCl3) δ ppm 2.30 (3H, s),
(31.5g, 46.5%) as a white solid. HPLC Purity: 93.3%. ESI-MS 2.55 (3H, s), 2.98 (4H, m), 3.65 (4H, m), 3.80 (3H, s), 5.22 (2H,
m/z (M+H+) Calcd 419.1, Obsd 419.3. 1H-NMR (600MHz, s), 6.11 (1H, s), 6.85–7.34 (7H, m), 7.57 (1H, s).
DMSO-d6) δ ppm 1.27 (3H, t, J=7.2Hz), 2.56 (3H, s), 3.70
Synthesis of Compound 6, (2E)-N-(2-Chloro-6-
(3H, s), 4.27 (2H, q, J=7.2Hz), 5.31 (2H, s), 6.89–6.90 (2H, methylphenyl)-2-({6-[4-(2-hydroxyethyl)piperazin-1-yl]-
m), 7.02 (1H, s), 7.42–7.43 (2H, m), 8.51 (1H, s). 13C-NMR 2-methylpyrimidin-4-yl}imino)-3-(4-methoxybenzyl)-2,3-
(150MHz, DMSO-d6) δ ppm 14.18, 25.10, 50.38, 55.05, 61.12, dihydro-1,3-thiazole-5-carboxamide-d4 To an ice-cooled
111.40, 113.30, 114.00, 127.90, 130.02, 135.75, 158.79, 159.01, solution of ethyl 2-bromoethanol-d4 (1.0eq.), compound 5
160.73, 161.48, 163.53, 166.31.
(9.47g, 1.5eq.) in DMF (60mL) was added dropwise Et3N
Synthesis of Compound 3, (2E)-2-[(6-Chloro-2-meth- (2.0eq.). The reaction mixture was heated to 60°C for 20h.
ylpyrimidin-4-yl)imino]-3-(4-methoxybenzyl)-2,3-di- The mixture was concentrated under vacuum, and the solid

880
Vol. 61, No. 8
was triturated successively with water and vigorously stirred
for 20min. The precipitate was collected by filtration and pu-
rified by column chromatography (CH2Cl2–CH3OH=10:1) to
give compound 6 (5.85g, 56.9%) as a light yellow solid. HPLC
Purity: 95.6%. ESI-MS m/z (M+H+) Calcd 612.2, Obsd 612.3.
1H-NMR (600MHz, CDCl3) δ ppm 2.30 (3H, s), 2.55 (3H, s),
2.65 (4H, m), 3.69–3.73 (4H, m), 3.80 (3H, s), 5.22 (2H, s), 6.12
(1H, s), 6.85–7.33 (7H, m), 7.57 (1H, s). 13C-NMR (150MHz,
DMSO-d6) δ ppm 18.28, 25.62, 43.67, 49.61, 52.90, 55.08,
58.52, 60.27, 91.74, 114.61, 116.40, 127.00, 128.18, 128.46,
129.04, 129.65, 130.79, 132.20, 133.20, 138.63, 158.96, 159.17,
159.26, 162.81, 163.32, 164.11.
Synthesis of Compound 7, 2-[4-(6-{[(2E)-5-{[(2-Chloro-
6-methylphenyl)amino]carbonyl}-3-(4-methoxybenzyl)-1,3-
thiazol-2(3H)-ylidene]amino}-2-methylpyrimidin-4-yl)-
piperazin-1-yl]ethyl N-(tert-Butoxycarbonyl)-l-leucinate-d4
To an ice-cooled solution of compound 6 (2.5g, 1.0eq.),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
(EDC·HCl, 1.5eq.), 1H-1,2,3-benzotriazol-1-ol (HOBT, 1.5eq.)
in CH2Cl2 (30mL) was added dropwise a solution of N-(tert-
butoxycarbonyl)-l-leucine (Boc-Leu-OH) in CH2Cl2 (1.2eq.,
10mL) and the solution was warmed to room temperature.
The mixture was stirred for 20h. The organic phase was
washed with 6–7% aq. NH3·H2O solution, dried, evaporated
and purified by column chromatography (petroleum ether–
ethyl acetate=1:2) to give compound 7 (2.18g, 64.7%) as a
Fig. 3. Mean Plasma Concentration–Time Profiles of Dasatinib and Da-
satinib-d4 after an Oral Dose of Equimolar Mixture of Dasatinib (5mg/
kg) and JLTN-d4 in Male Rhesus Monkeys (N=3)
Table 1. Mean Pharmacokinetic Parameters of Dasatinib and Dasatinib-
d4 in the Monkey
PK parameter
Dasatinib
Dasatinib-d4
Cmax (ng·mL−1)
Tmax (h)
38.9
4
54.9
4
MRTlast (h)
AUCall (ng·h·mL−1)
(RSD%)
6.8
173.5
(48.1%)
6.7
255.4
(47.8%)
light yellow solid. HPLC Purity: 97.9%. ESI-MS m/z (M+
1
H+) Calcd 825.4, Obsd 825.3. H-NMR (600MHz, CDCl3) δ 139.02, 156.84, 159.75, 160.42, 162.52, 164.12, 168.72.
ppm 0.95 (6H, m), 1.44 (9H, s), 1.55–1.67 (2H, m), 1.73 (1H,
Pharmacokinetics and Oral Bioavailability in Monkeys
m), 2.30 (3H, s), 2.55 (3H, s), 2.56–2.64 (4H, m), 3.56–3.71 The pharmacokinetics of equimolar mixture of Dasatinib
(4H, m), 3.80 (3H, s), 4.26–4.38 (1H, m), 4.89 (1H, m), 5.21 (5mg/kg) and JLTN-d4 were investigated in male Rhesus
(2H, s), 6.11 (1H, s), 6.86–7.32 (7H, m), 7.58 (1H, s). 13C NMR monkeys (approximately 4–5kg, N=3) following an oral dose
(150MHz, DMSO-d6) δ ppm 18.27, 21.36, 22.69, 24.24, 25.60, (capsule) by gavage in study design. The monkeys were fasted
27.94, 28.16, 43.66, 49.60, 51.96, 52.50, 55.08, 55.99, 61.69, overnight and fed 4h post dose. Blood samples were col-
78.10, 91.73, 114.15, 116.38, 127.00, 128.19, 128.45, 129.05, lected at 0, 5, 10, 30min, 1, 2, 3, 4, 6, 8, 12, 24 and 48h after
129.65, 130.80, 132.19, 133.19, 138.62, 155.50, 158.96, 159.14, oral dosing. Approximately 1mL of blood was collected in
159.26, 162.82, 163.25, 164.11, 172.99.
tubes containing ethylenediaminetetraacetic acid (EDTA) and
Synthesis of Compound 8, JLTN-d4, 2-(4-{6-[(5-{[(2-Chlo- plasma was obtained by centrifugation. Plasma samples were
ro-6-methylphenyl)amino]carbonyl}-1,3-thiazol-2-yl)- stored at −80°C until analysis. Samples were analyzed for
amino]-2-methylpyrimidin-4-yl}piperazin-1-yl)ethyl l-Leu- Dasatinib and Dasatinib-d4 by high performance liquid chro-
cinate Trihydrochloride-d4 To an ice-cooled solution of matography-tandem mass spectrometry (HPLC-MS-MS).15–18)
compound 7 (2.18g, 1.0eq.) in CH2Cl2 (30mL) was added
The pharmacokinetic parameters were computed by non-
dropwise CF3CO2H (30mL) and the solution was warmed compartmental analysis using Phoenix WinNonlin (Version
to room temperature. The solution was stirred for 48h, 6.3, Pharsight Corporation, CA, U.S.A.).
evaporated and the residual crude oil was added ethyl acetate
Mean pharmacokinetic parameters of Dasatinib and
(40mL). The solution was treated with hydrogen chloride– Dasatinib-d4 in the monkey are summarized in Table 1 and
ethyl acetate (5.0eq.) and vigorously stirred for 30min. The the plasma concentration time profiles are presented in Fig.
solid was filtered and dried in vacuo. The residue was sus- 3. The Tmax of Dasatinib and Dasatinib-d4 were both 4h. The
pended in DMF (35mL) and filtered. Then the filtrate was Cmax obtained directly from the concentration–time data were
added dropwise THF (70mL). The solid was filtered, washed 38.9ng·mL−1, 54.9ng·h·mL−1; the MRTlast were 6.8h, 6.7h; the
with ethyl acetate and dried in vacuo to obtain compound 8 AUCall were 173.5ng·h·mL−1 (RSD% 48.1%), 255.4ng·h·mL−1
(1.46g, 77.9%) as a white solid. HPLC Purity: 99.6%, isotopic (RSD% 47.8%); respectively. The geometric mean ratio (Dasa-
abundance: 98.58 atom %D. ESI-MS m/z (M+H+) Calcd 605.3, tinib-d4/Dasatinib) was 1.5 for AUCall and 1.4 for Cmax
.
1
Obsd 605.2. H-NMR (600MHz, DMSO-d6) δ ppm 0.91 (6H,
s), 1.63–1.74 (2H, m), 1.75–1.82 (1H, m), 2.24 (3H, s), 2.47 (3H,
Acknowledgment This work was supported by
a
s), 3.10–3.25 (2H, m), 3.50–3.61 (2H, m), 3.62–3.78 (2H, m), Grant from the National Program on Key Research Proj-
4.02–4.05 (1H, m), 4.26–4.46 (4H, m), 6.24 (2H, s), 7.20–7.40 ect of New Drug Innovation (No. 2008ZX09312-016,
(3H, m), 8.30 (1H, s), 8.80 (3H, s), 10.00 (1H, s), 11.49 (1H, s), 2012ZX09303006-002).
11.74 (1H, brs). 13C-NMR (150MHz, DMSO-d6) δ ppm 18.53,
22.20, 22.32, 23.99, 24.34, 41.51, 50.55, 50.91, 50.98, 53.98,
59.38, 84.40, 127.25, 128.58, 129.30, 132.63, 133.37, 133.51,
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