Novel Phenyl-Urea Derivatives As Dual-Target Ligands That Can Activate Both GK and Pparc

Acta Pharmaceutica Sinica B 2012;2(6):588–597

Institute of Materia Medica, Chinese Academy of Medical Sciences Chinese Pharmaceutical Association

Acta Pharmaceutica Sinica B

www.elsevier.com/locate/apsb www.sciencedirect.com

ORIGINAL ARTICLE Novel phenyl-urea derivatives as dual-target ligands that can activate both GK and PPARc

Lijian Zhanga,y, Kang Tiana,y, Yongqiang Lib,y, Lei Leic, Aifang Qina, Lijuan Zhanga, Hongrui Songa, Lianchao Huob, Lijing Zhangb, Xiaofeng Jinb, Zhufang Shenc, Zhiqiang Fengb,n aSchool of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China bInstitute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China cBeijing Key Laboratory of Active Substance Discovery and Druggability Evaluation, Institute of Materia Medica,Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China

Received 29 May 2012; revised 20 September 2012; accepted 9 October 2012

KEY WORDS Abstract A series of novel phenyl-urea derivatives which can simultaneously activate glucokinase (GK) and peroxisome proliferator-activated receptor g (PPARg) was designed and prepared, and Type 2 diabetes; GK activators; their activation of GK and PPARg was evaluated. The structure–activity relationships of these PPARg agonists; compounds are also described. Three compounds showed potent ability to activate both GK and Dual action PPARg. The possible binding mode of one of these compounds with GK and PPARg were predicted by molecular docking simulation.

nCorresponding author. Tel.: þ86 10 63189351. E-mail address: [email protected] (Zhiqiang Feng). yThese authors contributed equally to this work. Peer review under responsibility of Institute of Materia Medica, Chinese Academy of Medical Sciences and Chinese Pharmaceutical Association.

2211-3835 & 2012 Institute of Materia Medica, Chinese Academy of Medical Sciences and Chinese Pharmaceutical Association. Production and hosting by Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.apsb.2012.10.002 Novel phenyl-urea derivatives as dual-target ligands that can activate both GK and PPARg 589

1. Introduction ligand-activated transcription factors that offer a promising therapeutic approach for metabolic syndrome6. Activation of Type 2 diabetes (T2D), a complex and chronic disease PPARg by insulin-sensitizing thiazolidinedione (TZD) drugs7,8 characterized by hyperglycemia, is becoming more prevalent. leads to improve insulin action in muscle and liver. Several It was estimated that 366 million people will be afflicted with groups have described various synthetic small molecules as diabetes worldwide by 2030 and 95% of them are T2D1. There PPARg agonists (Fig. 2). mainly are three contributing factors leading to hyperglyce- Based on the above understanding of GK and PPARg,a mia, including reduction of insulin secretion from pancreatic drug able to simultaneously activate both GK and PPARg b-cells2, decreased tissue sensitivity to insulin and dysregulated would be expected to have a strong antihyperglycemia effect hepatic glucose metabolism. Although many antihyperglyce- by promoting insulin secretion, modulating hepatic glucose mia drugs have been developed, the therapeutic effects of the balance, and decreasing insulin resistance. commercially available single target drugs for T2D are Our design of compounds with dual action as both GK unsatisfactory as those drugs address only a part of the disease activators and PPARg agonists is based on the assembly of or a specific stage of T2D, and can induce drug resistance. ligand-based pharmacophores. We started from the fusion of a Therefore, it is necessary to develop multiple-target hypogly- GK activator 1 reported by Prosidion/OSI, and a PPARg cemic drugs with more widespread therapeutic effect. agonist 7 reported from Dainippon Sumitomo, to form the Glucokinase (GK), a glucose-phosphorylating enzyme and compound structure 11 with dual pharmacophores (Fig. 3). member of the hexokinase family,ismainlyexpressedintheliver Though these two compounds 1 and 7 are different in shape, and pancreatic b-cells. It is a potential therapeutic targets for they share the same disubstituted phenyl scaffold. We used the T2D3,4, as activation of GK not only promotes glycogen synthesis phenyl-urea structure of GK activator 1 as the parent compound but also enhances insulin secretion from pancreatic b-cells. Hence, backbone and added the pharmacophore of 7 and various the development of GK activators to lower blood glucose and nitrogen heteroaromatic groups to it, generating structure 12 normalize insulin secretion is a promising area of current research with three groups, R1,R2,andR3 that remained modifiable. for treating T2D. Several groups have described various synthetic Based upon this template, a series of phenyl-urea derivatives small molecules that act as GK activators (Fig. 1)5. were designed, prepared and evaluated for ability to activate GK Peroxisome proliferator-activated receptors (PPARs) exist and PPARg. Some of these compounds showed potent ability to as three isoforms (PPARa, PPARb and PPARg) and are activate both GK and PPARg simultaneously.

Figure 1 Structures of some GK activators reported and their pharmacophores (moieties in blue).

Figure 2 Structures of some PPARg agonists and their pharmacophores (moieties in blue). 590 Lijian Zhang et al.

Figure 3 Design strategy for GK/PPARg dual-target ligands.

Scheme 1 Synthesis of compounds 17a–f and 18a–f. Reagents and conditions: (a) K2CO3,DMF,801C, 6 h; (b) MeOH, 10% Pd-C, rt, 6–8 h; (c) R1Br, K2CO3, DMF, 80 1C, 4–8 h; (d) aldehyde, AcOH, EtOH, 70 1C, 4–8 h, and then NaBH4,601C, 10 h and (e) MeCN, 22 or 23, reﬂux.

Scheme 2 Synthesis of compounds 22 and 23. Reagents and conditions: (a) K2CO3, 2-aminothiazole or 2-amino-benzothiazole, MeCN, reﬂux.

2. Results and discussion 13 was O-alkylated with ethyl 2-bromoacetate in the presence of K2CO3 in DMF to give nitrobenzene derivatives 14. Aniline 2.1. Chemistry derivative 15 was obtained from reduction of 14, and the secondary amines 16a–f were prepared by the N-alkylation of The phenyl-urea derivatives were synthesized by the routes 15 with alkylbromide or aldehydes9. Subsequently, the con- shown in Schemes 1–3. Commercially available p-nitrophenol densation of 16a–f with the 4-nitrophenylcarbamate esters of Novel phenyl-urea derivatives as dual-target ligands that can activate both GK and PPARg 591

Scheme 3 Synthesis of compounds 20a–k. Reagents and conditions: (a) CDI, DMAP, DCM, reﬂux; (b) k2CO3, MeOH, H2O, 60 1C.

Table 1 GK activation effect of compounds 17a–f Table 2 GK activation effect of compounds 18a–f. and GKA22.

a a Compd. R1 Activation (fold) Compd. R1 Activation (fold)

17a Cyclopropylmethyl 1.2 18a Cyclopropylmethyl 0.8 17b Cyclohexylmethyl 1.0 18b Cyclohexylmethyl 1.0 17c n-Butyl 1.4 18c n-Butyl 1.3 17d n-Pentyl 1.8 18d n-Pentyl 1.3 17e i-Pentyl 1.2 18e i-Pentyl 1.2 17f 3-Methyl-2-butenyl 1.4 18f 3-Methyl-2-butenyl 0.8 GKA22 1.7 aGK activation (fold) was obtained at a Glc concentration of aGK activation (fold) was obtained at a Glc concentration of 4mM. 4 mM.

nitrogen. As shown in Table 1, the potency of compounds with heteroaromatic amines 22 or 23 produced the phenyl-urea acycloalkyl groups viz 17c–f, was superior to compounds with derivatives 17a–f or 18a–f (Scheme 1). Compounds 22 and 23 cycloalkyl groups viz 17a–b. The potency of compounds with were prepared starting from 4-nitrophenyl carbonochloridate straight-chain alkyl groups viz 17c–d, was slightly higher than (Scheme 2). The phenyl-urea derivatives 19a–k were prepared compounds with branched-chain alkyl groups viz 17e–f.Andthe by the three-component condensation of secondary amine 16d, potency of compound with n-pentyl group 17d was the strongest carbonyldiimidazole, and various 2-amino nitrogen heteroaro- in Table 1. matics10. Compounds 20a–k were obtained by the hydroliza- Subsequently, the activation of GK by the compounds 18a–f, bearing a 6-methylbenzothiazole ring replacing the thiazole ring of tion of 19a–k in a methanol–water–K2CO3 system (Scheme 3) 17a–f, was evaluated. As shown in Table 2, the potency of compounds 18a–f was obviously lower than that of the corre- 2.2. In vitro biological activity assay sponding compounds 17a–f. The similar structure–activity rela- tionship noted in Table 1 was observed for compounds 18a–f in The in vitro GK activity assay was performed at 37 1Cfor60min Table 2. in 96-well plates with a final volume of 150 mL reaction mixture AscanbeseenfromTables 1 and 2, replacing the thiazole ring containing 25 mM ATP, 1 mM NAD, 2 mM MgCl2, 5 mM DTT, of 17a–f with a 6-methylbenzothiazole ring had a detrimental 25 mM KCl, 100 mM Tris–HCl, 4.5 unit/mL glucose 6-phosphate effect on potency for GK activation, and the potency of 17a–f was dehydrogenase, 2 unit/mL recombinant GK, 4 mM glucose and notably increased when R1 was n-pentyl group 17d and 18d, test compound. After starting the reaction the increase in whether the nitrogen heteroaromatic moiety was thiazole or absorbance at 340 nm was monitored from 0 to 60 min with a 6-methylbenzothiazole. Subsequently, with R1 as a n-pentyl group, Bio-TEK microplate reader as a measure of GK activity. we explored the effect of a wide range nitrogen heteroaromatic The in vitro activation of GK by the target compounds were groups attached to the secondary urea nitrogen on GK activation evaluated at a concentration 10 mM, and GKA22, a GK efficiency. activator developed by the AstraZeneca11, was used as positive As shown in Table 3, the compound 19a with a pyridine control. The compounds that showed greatest ability to activate ring and 17d with a thiazole ring were equipotent in GK GK were selected for further study and for their ability to activation. The substitution of simple thiazole group led to activate transcription by PPARg in a cell-based assay. obvious reduction of potency, for example 19g–i. And when We first estimated the ability of compounds 17a–f to activate R3 was H instead of the ethyl, the same substitution of GK; these bear a thiazole ring attached to the secondary urea thiazole group analogs 20e–g, also displayed less potency than 592 Lijian Zhang et al.

Table 3 GK activation effect of the compounds 19a–k Table 3 (continued ) and 20a–k.

Activation Compd. R R 3 2 (fold)a Activation Compd. R R 3 2 (fold)a

20c H 1.3 19a Et 1.8

20d H 1.1 19b Et 1.4

19c Et 0.8 20e H 1.6

20f H 1.4 19d Et 1.7

20g H 1.3 19e Et 1.4

20h H 1.2 19f Et 1.2

20i H 1.5 19g Et 1.6

20j H 1.7 19h Et 1.5

20k H 1.6

19i Et 1.6

aGK activation (fold) was obtained at a Glc concentration of 4mM.

19j Et 1.5 the corresponding compounds 17d and 19g–h.WhenR2 was a pyridine derivates group12,13, the simple substitution of pyridine group, for example 19b–f led to significant reduction in or loss of potency, except compound 19d which contains a 5- 19k Et 1.3 methoxycarbonyl pyridine moiety showing a GK activation fold of 1.7, similar to the non-substituted pyridine group analog 19a. Interestingly, when R3 was H the GK activation fold of the high potency compound 19a was reduced, the corresponding com- 20a H 1.6 pound 20a was 1.6, and the low potency compounds 19b–c were increased, with the potency of the corresponding compounds 20b–c was 1.7 and 1.3, respectively. In addition, when R2 was a 20b H 1.7 benzothiazole derivates group the simply C-6 substitution of a benzothiazole group also led to slight reduction of potency, for Novel phenyl-urea derivatives as dual-target ligands that can activate both GK and PPARg 593 example 18d and 19k.ButwhenR was H replacing the ethyl, 3 Table 4 Activation of selected compounds to GK and the same substitution of benzothiazole group analogs 20j–k, PPARg. displayed remarkably improved potency, with GK activation fold of 1.7 and 1.6, respectively, as compared to that of Compd. EC1.5 (mM)a PPARg (%)b corresponding compounds 18d and 19k. Based on the ability of the targeted compounds to activate 17d 79.17 44.6 GK, seven potential candidates (activation foldZ1.6) were 19a 94.70 41.5 selected for further evaluation in the GK enzyme assay 19d 5.35 43.4 (EC1.5) and a cell-based PPARg transcription assay. As 19i 68.89 64.1 20e 32.12 50.9 shown in Table 4, compounds 19d, 20j and 20k not only were 20j 7.52 54.0 potent GK activators with EC1.5 values of 5.35, 7.52 and 20k 6.85 51.2 6.85 mM, respectively, similar to that of GKA22, but also GKA22 3.50 – displayed the activity to activate PPARg transcription with a potency in the range of 43–54% as compared to rosiglitazone. aEC1.5 means the concentration of compounds required to We also predicted the binding mode for these compounds in the increase enzyme activity by 50%. bThe effect of tested compounds on activity of PPARg ligand-binding domains of GK and PPARg respectively by using 6 the CDOCKER docking program. As shown in Fig. 4A, com- transcription at 10 M was evaluated by luciferase reporter gene assay and presented as activity relative to rosiglitazone. pound 19d binds at the allosteric site of GK similar to that of other The activity of rosiglitazone is assigned as 100%. reported complexes14,15, and forms a bidentate hydrogen bond by the pyridine N atom and the NH group of the amide with the backbone NH and carbonyl oxygen of Arg63 on GK. This may explain its potent ability to activate GK. Meanwhile, Fig. 4B represents the main interactions between compound 19d and 14.4 mmol) and ethyl bromoacetate (2.7 g, 16.2 mmol) in PPARg. Compound 19d does not form a network of hydrogen DMF (20 mL), the reaction mixture was heated at 80 1C for bonds with the side chains of Tyr473, His449, Ser289 and His323 6 h, then cooled to room temperature and poured into 100 mL in the polar cavity of the PPARg as rosiglitazone does, but showed water, extracted with ethyl acetate several times. The com- H-bond interactions with Cys285, Ser289, Tyr327 and Ser342. The bined organic layers were washed with brine, dried on absence of strong interactions may explain the significant decrease anhydrous Na2SO4, filtered and evaporated under reduced in the activation of PPARg transcription. pressure to afford 3.08 g of compound 14, yield 95%.

4.2. Ethyl 2-(4-aminophenoxy) acetate (15). 3. Conclusion A suspension of ethyl 2-(4-nitrophenoxy) acetate 14 (450 mg, In summary, a series of phenyl-urea derivatives were designed, 2 mmol) and Pd-C (45 mg) in methanol (15 mL) was treated prepared and evaluated for their ability to activate GK and with H at atmosphere pressure and room temperature for 6– PPARg. Some of these compounds, for example 19d, 20j and 2 8 h. The mixture was filtered and concentrated under reduced 20k, showed potent activity towards both GK and PPARg,and pressure to give 378 mg of 15, yield 97%. the possible binding modes of compounds 19d with GK and PPARg were predicted by in silico molecular docking. These dual actions towards GK and PPARg may produce a more wide- 4.3. General procedure for the preparation of compounds spread therapeutic effect on the hyperglycemia associated with 16a–f. T2D. The discovery of compounds that act on both targets gives a new approach to the design of anti-diabetic agents. Method 1: the corresponding alkyl bromides (1 equiv.) was slowly added to a stirred solution of ethyl 2-(4-aminophenoxy) acetate 15 (1 equiv.) and anhydrous potassium carbonate 4. Experimental (0.5 equiv.) in dry DMF, and then heated at 80 1C until the reaction completed. The mixture was diluted with ethyl acetate All reagents and chemicals were obtained from commercial and washed with water and brine successively, dried over suppliers and used as received. Reaction progress was mon- anhydrous Na2SO4, filtered and concentrated under reduced itored by thin-layer chromatography on Qindao silica gel 60 pressure. The residue was purified by column chromatography F-254 with UV detection. Flash column chromatography was with PE/EtOA as eluent. performed on silica gel 60 (200–300 mesh) from Qindao 1 Method 2: several drops AcOH and the corresponding Haiyang Chemical Factory. H NMR spectra were recorded aldehyde (1 equiv.) were added to a stirred solution of ethyl 2- on a Mercury-300 spectrometer with Chemical shifts in (4-aminophenoxy) acetate 15 (1 equiv.) in MeOH, and then ppm (d) relative to the solvent signal and coupling constant heated to 70 1C. When the reaction was completed, NaBH4(3 (J) values in Hz. HPLC-MS analysis was performed on a equiv.) was added and reacted at 60 1C for 10 h, then Thermo Finnigan LCQ-Advantage mass spectrometer. evaporated the solvent under reduced pressure. The residue was diluted with ethyl acetate and washed with water and 4.1. Ethyl 2-(4-nitrophenoxy) acetate (14). brine successively, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude product Anhydrous potassium carbonate (2.5 g, 17.4 mmol) in por- obtained was further purified by column chromatography tions was added to a stirred solution of 4-nitrophenol 13 (2 g, with PE/EtOA as eluent. 594 Lijian Zhang et al.

4.4. 4-Nitrophenyl-thiazol-2-yl-carbamate (22) and acidiﬁed with 1 M HCl, and ﬁltrated, recrystallized to give the 4-nitrophenyl-6-methylbenzo[d] thiazol-2-yl-carbamate (23). target compounds 20a–k.

4-Nitrophenyl chloroformate (1.2 equiv.) was added to a 4.7. Characterization of new products stirred mixture of thiazol-2-amine (1 equiv.) and anhydrous potassium carbonate (0.6 equiv.) in MeCN, then refluxed. 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(cyclopropylmethyl)- The reaction mixture was concentrated and diluted with ethyl 1 3-(thiazol-2-yl)urea(17a): H NMR (DMSO-d6, 300 MHz), d: acetate, washed with water several times. The organic phase 9.68 (s, 1H, CONH), 7.23 (m, 3H, ArH), 6.97 (m, 3H, ArH), was dried with anhydrous Na2SO4, then filtered, concen- 4.80 (s, 1H, COCH2), 4.18 (q, 2H, J¼6.9 Hz, CH2), 3.52 trated and used in the next step. Use the 6-methylbenzo[d] (d, 2H, J¼6.6 Hz, CH2), 1.22 (t, 3H, J¼6.9 Hz, CH3), thiazol-2-amine instead of thiazol-2-amine and the same 0.89–0.94 (m, 1H, CH¼), 0.34–0.40 (m, 2H, CH2), 0.04– method to give 4-nitrophenyl 6-methylbenzo[d]thiazol-2-yl- þ 0.09 (m, 2H, CH2); m/z 376 [MþH] ; HR-MS calculated for carbamate (23). þ C18H21N3O4S: m/z 376.1326 [MþH] , found 376.1326. 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(cyclohexylmethyl)- 1 3-(thiazol-2-yl)urea(17b): H NMR (DMSO-d6, 300 MHz), d: 4.5. General procedure for the preparation of compounds 9.60 (s, 1H, ¼CHNH–), 7.25 (d, 1H, J¼3.3 Hz, ArH), 7.20 (d, 17a–f and 18a–f. 2H, J¼8.7 Hz, ArH), 6.95–6.98 (m, 3H, ArH), 4.80 (s, 1H, COCH2), 4.18 (q, 2H, J¼7.2 Hz, CH2), 3.53 (d, 2H, Intermediates 22 or 23 (1 equiv.) was added to a solution of J¼6.9 Hz, CH2), 1.56–1.65 (m, 4H, CH22), 1.39 (m, 16a–f (1.1 equiv.) in MeCN and refluxed. After the reaction 1H, CH¼), 1.22 (t, 3H, J¼7.2 Hz, CH3), 1.09–1.18 (m, 4H, þ was completed, the reaction mixture was cooled to r.t. and the CH2 2), 0.88–0.93 (m, 2H, CH2); m/z 418 [MþH] ;HR- þ solvent was evaporated under reduced pressure. The residue MS calculated for C21H27N3O4S: m/z 418.179 [MþH] , found was then diluted with dichloromethane and washed with water 418.1795. and brine, dried by anhydrous Na2SO4, filtered and concen- 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-butyl)-3-(thiazol-2- 1 trated under reduced pressure. The rude product was purified yl)urea (17c): H NMR (DMSO-d6, 300 MHz), d: 7.27 (d, 1H, by column chromatography to give 17a–f and 18a–f. ¼CHNH), 7.23 (d, 2H, J¼3.6 Hz, ArH), 7.00 (d, 3H, J¼ 3.6 Hz, ArH), 4.81 (s, 2H, COCH2), 4.23 (q, 2H, J¼7.2 Hz, OCH2), 3.66 (t, 2H, J¼6.9 Hz, CH2), 1.39 (q, 2H, 4.6. General procedure for the preparation of compounds J¼6.9 Hz, CH2), 1.301.15 (m, 5H, CH2CH3), 0.91 (t, þ 19a–k and 20a–k. 3H, J¼7.2 Hz, CH3); m/z 377.1 [MþH] ; HR-MS calcu- þ lated for C18H23N3O4S: m/z 378.1482 [MþH] , found The hetero-aromatic amine (1 equiv.) and 4-dimethylamino- 378.1058. pyridine (0.05 equiv.) were addedtoastirredsolutionof1,10- 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(thiazol- 1 carbonyldiimidazole (1.1 equiv.) in anhydrous dichloromethane, 2-yl)urea (17d): H NMR (DMSO-d6, 300 MHz), d:9.81 and the reaction was refluxed for 3 h, then cooled to 0 1C, and the (s, 1H, CONH), 7.25 (d, 1H, J¼3.9 Hz, ArH), 7.20 (d, 2H, ethyl 2-[4-(pentylmino)phenoxy]acetate (16d)wasaddedinpor- J¼8.7 Hz, ArH), 6.95–6.98 (m, 3H, ArH), 4.80 (s, 2H, tions, then refluxed until reaction was completed. The mixture was COCH2), 4.18 (q, 2H, J¼6.9 Hz, CH2), 3.62 (t, 2H, diluted with dichloromethane, and washed with water and brine, J¼7.2 Hz, CH2), 1.40–1.45 (m, 2H, CH2), 1.20–1.24 driedonanhydrousNa2SO4, filtered and concentrated under (m, 7H, CH22, CH3), 0.83 (t, 3H, J¼6.9 Hz, -CH3); m/z þ reduced pressure. The residue was purified by column chromato- 392.2 (MþH) ; HR-MS calculated for C19H25N3O4S: m/z graphy to afford compounds 19a–k. 392.1632 [MþH]þ, found 392.1639. Potassium carbonate (2 equiv.) was added to a solution of the 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(i-pentyl)-3-(thia- 1 compounds 19a–k (1 equiv.) in methanol and water (5:1, v/v). zol-2-yl)urea (17e): H NMR (DMSO-d6, 300 MHz), d: 9.63 The mixture was heated at 60 1C until the reaction finished. The (s, 1H, CONH), 7.247.18 (m, 3H, ArH), 6.97 (m, 2H, solvent was evaporated under reduced pressure. The residue was ArH), 4.80 (s, 2H, COCH2), 4.18 (q, 2H, J¼7.2 Hz,

Fig. 4 Calculated binding mode of hit compound 19d. The structures of GK (A) and PPARg (B) are displayed as cartoon in off-white. Selected residues are shown as thin sticks (carbons in purple, oxygens in red and nitrogen in blue). Coloring for the ligand is as follows: carbons in green, oxygens in red, nitrogens in blue and sulfur in orange. Predicted H-bonds are shown as white thin lines. Novel phenyl-urea derivatives as dual-target ligands that can activate both GK and PPARg 595

þ CH2), 3.65 (t, 2H, J¼7.2 Hz, CH2), 1.581.51 (m, 1H, 456.2 [MþH] ; HR-MS calculated for C24H29N3O4S: m/z þ CH¼), 1.28–1.35 (m, 2H, CH2), 1.21 (t, 3H, J¼6.9 Hz, 456.1944 [MþH] , found 456.1952. þ CH3), 0.83 (d, 6H, J¼6.6 Hz, CH3 2); m/z 392 [MþH] ; 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(3-methyl-2-bute- þ 1 HR-MS calculated for C19H25N3O4S: m/z 392.1630 [MþH] , nyl)-3-(6-methylbenzo[d] thiazol-2-yl)urea (18f): H NMR found 392.0625. (DMSO-d6, 300 MHz), d: 10.04 (s, 1H, CONH), 7.58 (s, 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(3-methyl-2-butenyl)- 1H, ArH), 7.28–7.11 (m, 4H, ArH), 6.93 (d, 2H, J¼8.4 Hz, 1 3-(thiazol-2-yl)urea (17f): H NMR (DMSO-d6, 300 MHz), d: ArH), 5.22 (t, 1H, J¼6.9 Hz, CH¼), 4.79 (s, 2H, 9.70 (s, 1H, CONH), 7.24 (d, 1H, J¼3.6 Hz, ArH), 7.16 COCH2), 4.14–4.27 (m, 4H, CH22), 2.34 (s, 3H, (d, 2H, J¼8.7 Hz, ArH), 6.94 (m, 3H, ArH), 5.20 (t, 1H, CH3), 1.62 (s, 3H, CH3), 1.44 (s, 3H, CH3), 1.20 (t, 3H, þ J¼6.9 Hz, CH¼), 4.79 (s, 2H, COCH2), 4.14–4.24 (m, J¼6.9 Hz, CH3); m/z 454 [MþH] ; HR-MS calculated for þ 4H, CH22), 1.62 (s, 3H, CH3), 1.42 (s, 3H, CH3), C24H27N3O4S: m/z 454.1792 [MþH] , found 454.1795. þ 1.21 (t, 3H, J¼7.2 Hz, CH3); m/z 390 [MþH] ; HR-MS 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(pyri- þ 1 calculated for C19H23N3O4S: m/z 390.1481 [MþH] , found dine-2-yl)urea (19a): H NMR (DMSO-d6, 300 MHz), d: 390.1482. 8.07 (m, 1H, ArH), 7.94 (d, 1H, J¼8.7 Hz ArH), 7.64–7.73 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(cyclopropylmet (m, 1H, ArH), 7.32 (d, 2H, J¼8.7 Hz, ArH), 7.06 (d, 2H, hyl)-3-(6-methylbenzo[d] thiazol-2-yl)urea (18a): 1H NMR J¼8.7 Hz, ArH), 7.04 (s, 1H, CONH) 6.93–6.97 (m, 1H, (DMSO-d6, 300 MHz), d: 10.11 (s, 1H, CONH), 7.58 (s, ArH), 4.84 (s, 2H, COCH2), 4.19 (t, 2H, J¼6.6 Hz, 1H, ArH), 7.11–7.25 (m, 4H, ArH), 6.96 (d, 2H, J¼8.7 Hz, CH2), 3.60 (t, 2H, J¼7.2 Hz, CH2), 1.40–1.45 (m, ArH), 4.80 (s, 2H, COCH2), 4.18 (q, 2H, J¼7.2 Hz, 2H, CH2), 1.21–1.40 (m, 4H, CH22), 1.19–1.32 (m, þ CH2), 3.53 (d, 2H, J¼6.9 Hz, CH2), 2.34 (s, 1H, 3H, CH3), 0.83 (t, 3H, J¼6.9 Hz, CH3); m/z 386 [MþH] ; þ CH3), 1.22 (t, 3H, J¼6.9 Hz, CH3), 0.89–0.94 (m, 1H, HR-MS calculated for C21H27N3O4: m/z 386.2063 [MþH] , CH¼), 0.37–0.41 (m, 2H, CH2), 0.04–0.09 (m, 2H, – found 386.2074. þ CH2–); m/z 440.2 [MþH] ; HR-MS calculated for 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(5-methyl- þ 1 C23H25N3O4S: m/z 440.1640 [MþH] , found 440.1639. pyridine-2-yl)urea (19b): HNMR(DMSO-d6,300MHz),d:7.90 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(cyclohexylmethyl)-3- (m, 1H, ArH), 7.84 (d, 1H, J¼8.7 Hz, ArH), 7.52 (dd, 1H, 1 (6-methylbenzo[d] thiazol-2-yl)urea (18b): HNMR(DMSO-d6, J1¼2.1 Hz, J2¼8.4 Hz, ArH), 7.30 (d, 2H, J¼8.7 Hz, ArH), 7.05 300 MHz), d:9.93(s,1H,CONH), 7.56 (s, 1H, ArH), (d, 2H, J¼8.7 Hz, ArH), 6.97 (s, 1H, CONH), 4.84 (s, 2H, 7.12–7.20 (m, 3H, ArH), 6.95–6.97 (m, 3H, ArH), 4.80 COCH2), 4.18 (q, 2H, J¼6.9 Hz, CH2), 3.60 (t, 2H, (s, 2H, COCH2), 4.19 (q, 2H, J¼7.2 Hz, CH2), 3.56 J¼6.9 Hz, CH2), 2.16 (s, 3H, CH3), 1.40–1.44 (m, 2H, (d, 2H, J¼6.9 Hz, CH2), 2.34 (s, 1H, CH3), 1.56–1.66 CH2), 1.251.18 (m, 7H, CH22, CH3), 0.82 (t, 3H, þ (m, 4H, CH22), 1.41 (m, 1H, CH¼), 1.22 (t, 3H, J¼6.9 Hz, CH3); m/z 400 [MþH] ; HR-MS calculated for þ J¼6.9 Hz, CH3), 1.09–1.18 (m, 4H, CH22), 0.88–0.93 C22H29N3O4: m/z 400.222 [MþH] , found 400.2231. þ (m, 2H, CH2); m/z 482.2 [MþH] ;HR-MScalculatedfor 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(5-chlo þ 1 C26H31N3O4S: m/z 482.2115 [MþH] , found 482.2108. ropyridine-2-yl)urea (19c): H NMR (DMSO-d6, 300 MHz), d: 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-butyl)-3-(6-methy 8.18–8.22 (dd, 1H, J1 ¼0.6 Hz, J2 ¼2.4 Hz, ArH), 7.96 (d, 1H, 1 lbenzo[d]thiazol-2-yl) urea (18c): H NMR (DMSO-d6, J¼9.0 Hz, ArH), 7.81 (dd, 1H, J1 ¼2.4 Hz, J2 ¼9.0 Hz, ArH), 300 MHz), d: 7.58 (s, 1H, ArH), 7.29 (s, 1H, ArH), 7.22 7.29–7.32 (m, 3H, ArH, CONH), 7.04 (d, 2H, J¼9.0 Hz, (d, 2H, J¼3.6 Hz, ArH), 7.14 (d, 1H, J¼3.6 Hz, ArH), 4.80 ArH), 4.83 (s, 2H, COCH2), 4.18 (q, 2H, J¼6.9 Hz, (s, 2H, COCH2), 4.22 (q, 2H, J¼6.9 Hz, CH2), 2.34 CH2), 3.60 (t, 2H, J¼7.2 Hz, CH2), 1.40–1.44 (m, (s, 3H, CH3), 1.43 (t, 2H, J¼6.9 Hz, CH2), 1.301.20 2H, CH2), 1.25–1.18 (m, 7H, CH22, CH3), 0.83 (t, þ (m, 5H, CH2CH3), 0.87 (t, 3H, J¼6.9 Hz, CH3); m/z 441.2 3H, J¼6.6 Hz, CH3); m/z 420 [MþH] ; HR-MS calculated þ þ [MþH] ; HR-MS calculated for C23H27N3O4S: m/z 422.1795 for C22H26ClN3O4: m/z 420.1679 [MþH] , found 460.1685. [MþH]þ, found 422.1804. 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(5- 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3- methoxylformylpyridine-2-yl) urea (19d): 1H NMR (DMSO- 1 (6-methylbenzo[d]thiazol-2-yl) urea (18d): H NMR d6, 300 MHz), d: 8.63 (d, 1H, J¼2.1 Hz, ArH), 8.18–8.22 (dd, (DMSO-d6, 300 MHz), d: 10.08 (s, 1H, CONH), 7.58 (s, 1H, J1 ¼2.1 Hz, J2 ¼8.7 Hz, ArH), 8.06 (d, 1H, J¼8.7 Hz, 1H, ArH), 7.29 (s, 1H, ArH), 7.20 (d, 2H, J¼8.7 Hz, ArH), ArH), 7.60 (s, 1H, CONH), 7.32 (d, 2H, J¼8.7 Hz, ArH), 7.12 (d, 1H, J¼8.4 Hz, ArH), 6.96 (d, 2H, J¼9 Hz, ArH), 7.05 (d, 2H, J¼8.7 Hz, ArH), 4.84 (s, 2H, COCH2), 4.14– 4.80 (s, 2H, COCH2), 4.18 (q, 2H, J¼6.9 Hz, CH2), 4.21 (q, 2H, J¼6.9 Hz, CH2), 3.81 (s, 3H, OCH3), 3.62 3.65 (t, 2H, J¼7.2 Hz, CH2), 2.34 (s, 3H, CH3), 1.44 (t, 2H, J¼7.5 Hz, CH2), 1.41–1.46 (m, 2H, CH2), 1.14– (m, 2H, CH2), 1.14–1.29 (m, 7H, CH2 2, CH3), 0.83 1.26 (m, 7H, CH22, CH3), 0.83 (t, 3H, J¼6.9 Hz, þ þ (t, 3H, J¼6.9 Hz, CH3); m/z 456 [MþH] ; HR-MS calcu- CH3); m/z 444.2 [MþH] ; HR-MS calculated for þ þ lated for C24H29N3O4S: m/z 456.1955 [MþH] , found C23H29N3O6: m/z 444.2114 [MþH] , found 444.2129. 456.1952. 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(5- 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(i-pentyl)-3-(6-me- ethoxylformylpyridine-2-yl) urea (19e): 1H NMR (DMSO- 1 thylbenzo[d]thiazol-2-yl) urea (18e): H NMR (DMSO-d6, d6, 300 MHz), d: 8.62 (s, 1H, ArH), 8.33 (d, 1H, J¼8.7 Hz, 300 MHz), d: 7.58 (s, 1H, ArH), 7.18–7.28 (m, 3H, ArH), 7.12 ArH), 8.06 (d, 1H, J¼9.0 Hz, ArH), 7.60 (s, 1H, CONH), (d, 1H, J¼7.5 Hz, ArH), 6.96 (d, 2H, J¼8.7 Hz, ArH), 4.80 7.32 (d, 2H, J¼8.4 Hz, ArH), 7.05 (d, 2H, J¼8.4 Hz, ArH), (s, 2H, COCH2), 4.17 (q, 2H, J¼7.2 Hz, CH2), 3.68 (t, 4.84 (s, 2H, COCH2), 4.24–4.31 (q, 2H, J¼7.2 Hz, 2H, J¼7.2 Hz, CH2), 2.34 (s, 3H, CH3), 1.48–1.59 (m, CH2), 4.14–4.21 (q, 2H, J¼6.9 Hz, CH2), 3.62 (t, 2H, 1H, CH¼), 1.371.30 (m, 2H, CH2), 1.22 (t, 3H, J¼6.6 Hz, CH2), 1.44 (m, 2H, CH2), 1.18–1.30 (m, J¼6.9 Hz, CH3), 0.83 (d, 6H, J¼6.9 Hz, CH3 2); m/z 10H, CH22, CH3 2), 0.83 (t, 3H, J¼6.9 Hz, CH3); 596 Lijian Zhang et al.

þ m/z 458.2 [MþH] ; HR-MS calculated for C24H31N3O6: m/z 1-[4-(2-Hydroxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3- þ 1 458.228 [MþH] , found 458.2286. (pyridine-2-yl)urea (20a): H NMR (DMSO-d6, 300 MHz), 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(5-iso d: 8.07 (m, 1H, ArH), 7.94 (d, 1H, J¼8.7 Hz, ArH), 7.67–7.72 1 propoxylformylpyridine-2-yl)urea (19f): H NMR (DMSO-d6, (m, 1H, ArH), 7.29 (d, 2H, J¼8.7 Hz, ArH), 6.98 (s, 1H, 300 MHz), d: 8.60 (d, 1H, J¼1.5 Hz, ArH), 8.16–8.20 (dd, 1H, CONH), 6.92–7.02 (m, 3H, ArH), 4.57 (s, 2H, J1 ¼2.1 Hz, J2 ¼8.7 Hz, ArH), 8.05 (d, 1H, J¼8.7 Hz, ArH), COCH2), 3.59 (t, 2H, J¼7.5 Hz, CH2), 1.43 (m, 2H, 7.60 (s, 1H, CONH), 7.32 (d, 2H, J¼8.7 Hz, ArH), 7.05 CH2), 1.23–1.25 (m, 4H, CH22), 0.83 (t, 3H, J¼6.9 þ (d, 2H, J¼8.7 Hz, ArH), 5.10 (m, 1H, CH¼), 4.84 Hz, CH3); m/z 358 [MþH] ; HR-MS calculated for þ (s, 2H, COCH2), 4.14–4.21 (q, 2H, J¼6.9 Hz, CH2), C19H23N3O4: m/z 358.1763 [MþH] , found 358.1761. 3.62 (t, 2H, J¼7.5 Hz, CH2), 1.44 (m, 2H, CH2), 1.14– 1-[4-(2-Hydroxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(5- 1 1.26 (m, 13H, CH22, CH3 3), 0.83 (t, 3H, J¼6.9 methylpyridine-2-yl)urea (20b): H NMR (DMSO-d6, þ Hz, CH3); m/z 472.2 [MþH] ; HR-MS calculated for 300 MHz), d: 13.03 (s, 1H, OH), 7.90 (m, 1H, ArH), 7.83 þ C25H33N3O6: m/z 472.2445 [MþH] , found 472.2442. (d, 1H, J¼8.4 Hz, ArH), 7.53 (dd, 1H, J1 ¼2.1 Hz, J 2 ¼ 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(5-meth 8.4 Hz, ArH), 7.30 (d, 2H, J¼9.0 Hz, ArH), 7.01–7.04 (m, 3H, 1 ylthiazol-2-yl)urea (19g): H NMR (DMSO-d6, 300 MHz), d: ArH, CONH), 4.73 (s, 2H, COCH2), 3.59 (t, 2H, 9.63 (s, 1H, CONH), 7.18 (d, 2H, J¼8.7 Hz, ArH), 6.95 J¼7.2 Hz, CH2), 2.16 (s, 1H, CH3), 1.40–1.44 (m, 2H, (d, 2H, J¼8.7 Hz, ArH), 6.48 (s, 1H, ArH), 4.79 CH2), 1.24 (m, 4H, CH22), 0.82 (t, 3H, J¼6.6 Hz, þ (s, 2H, COCH2), 4.17 (q, 2H, J¼6.9 Hz, CH2), 3.61 CH3); m/z 372.2 [MþH] ; HR-MS calculated for þ (t, 2H, J¼7.5 Hz, CH2), 2.12 (s, 3H, CH3) 1.40–1.50 C20H25N3O4: m/z 372.191 [MþH] , found 372.1918. (m, 2H, CH2), 1.241.14 (m, 7H, CH22, CH3), 1-[4-(2-Hydroxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(5- þ 1 0.80–0.84 (t, 3H, J¼6.9 Hz, CH3); m/z 406.2 [MþH] ;HR- chloropyridine-2-yl)urea (20c): H NMR (DMSO-d6, þ MS calculated for C20H27N3O4S: m/z 406.1786 [MþH] , found 300 MHz), d: 13.05 (s, 1H, OH), 8.13 (d, J¼2.7 Hz, ArH), 406.1795. 7.97 (d, 1H, J¼9.0 Hz, ArH), 7.82 (dd, 1H, J1 ¼2.7 Hz, 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3- J2 ¼9.0 Hz, ArH), 7.29–7.32 (m, 3H, ArH, CONH), 7.02 1 (5-chlorothiazol-2-yl)urea (19h): H NMR (DMSO-d6, (d, 2H, J¼8.7 Hz, ArH), 4.73 (s, 2H, COCH2), 3.60 (t, 2H, 300 MHz), d: 10.34 (s, 1H, CONH), 7.40 (s, 1H, ArH), J¼7.2 Hz, CH2), 1.43 (m, 2H, CH2), 1.24 (m, 4H, 7.20 (d, 2H, J¼8.7 Hz, ArH), 6.81 (d, 2H, J¼8.7 Hz, ArH), CH22), 0.83 (t, 3H, J¼6.6 Hz, CH3); m/z 392 þ 4.80 (s, 2H, COCH2), 4.17 (q, 2H, J¼6.9 Hz, CH2), [MþH] ; HR-MS calculated for C19H22ClN3O4: m/z þ 3.60 (t, 2H, J¼7.5 Hz, CH2), 1.40–1.50 (m, 2H, CH2), 392.1373 [MþH] , found 392.1372. 1.24–1.33 (m, 7H, CH22, CH3), 0.80–0.84 1-[4-(2-Hydroxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(5- þ 1 (t, 3H, J¼6.9 Hz, CH3); m/z 426.1 [MþH] ; HR-MS hydroxylformylpyridine-2-yl)urea (20d): H NMR (DMSO-d6, þ calculated for C19H24ClN3O4S: m/z 426.1245 [MþH] , found 300 MHz), d: 13.03 (s, 2H, OH 2), 8.64 (d, 1H, J¼2.1 Hz, 426.1249. ArH), 8.20 (m, 1H, ArH), 8.05 (d, 1H, J¼8.7 Hz, ArH), 7.52 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3- (s, 1H, CONH), 7.32 (d, 2H, J¼8.7 Hz, ArH), 7.03 (d, 2H, 1 (5-ethoxylformylthiazol-2-yl)urea (19i): H NMR J¼8.7 Hz, ArH), 4.74 (s, 2H, COCH2), 3.62 (t, 2H, (DMSO-d6, 300 MHz), d: 10.54 (s, 1H, CONH), 7.85 (s, J¼6.9 Hz, CH2), 1.44 (m, 2H, CH2), 1.24 (m, 4H, 1H, ArH), 7.20 (d, 2H, J¼8.7 Hz, ArH), 6.97 (d, 2H, J¼8.7 CH22), 0.83 (t, 3H, J¼6.9 Hz, CH3); m/z 402 þ Hz, ArH), 4.80 (s, 2H, COCH2), 4.14–4.26 (m, 4H, [MþH] ; HR-MS calculated for C20H23N3O6: m/z 402.1669 þ CH22), 3.61 (t, 2H, J¼7.2 Hz, CH2), 1.42 (m, 2H, [MþH] , found 402.1660. CH2), 1.14–1.30 (m, 10H, CH22, CH3 2), 0.80– 1-[4-(2-Hydroxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(thia- þ 1 0.84 (t, 3H, J¼6.9 Hz, CH3); m/z 464.2 [MþH] ; HR-MS zol-2-yl)urea (20e): H NMR (DMSO-d6, 300 MHz), d: 7.25 þ calculated for C22H29N3O6S: m/z 464.1855 [MþH] , found (d, 1H, J¼3.9 Hz, ArH), 7.20 (d, 2H, J¼8.7 Hz ArH), 6.94– 464.1850. 6.98 (m, 3H, ArH), 4.70 (s, 2H, COCH2), 3.62 (t, 2H, 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(benzo J¼6.9 Hz, CH2), 1.42 (m, 2H, CH2), 1.21–1.24 (m, 4H, 1 [d]thiazol-2-yl)urea (19j): H NMR (DMSO-d6,300MHz),d: CH22), 0.82 (t, 3H, J¼6.9 Hz, CH3); m/z 364 þ 10.24 (s, 1H, CONH), 7.78 (d, 1H, J¼7.5 Hz, ArH), [MþH] ; HR-MS calculated for C17H21N3O4S: m/z 364.1329 7.397.29 (m, 2H, ArH), 7.227.15 (m, 3H, ArH), 6.96 (d, [MþH]þ, found 364.1326. 2H, J¼8.4 Hz, ArH), 4.80 (s, 2H, COCH2), 4.15–4.22 (q, 1-[4-(2-Hydroxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3- 1 2H, J¼6.9 Hz, CH2), 3.66 (t, 2H, J¼7.2 Hz, CH2), 1.44 (5-methylthiazol-2-yl)urea (20f): H NMR (DMSO-d6, (m, 2H, CH2), 1.16–1.29 (m, 7H, CH22, CH3), 0.83 300 MHz), d: 7.18 (d, 2H, J¼8.7 Hz, ArH), 6.93 (d, 2H, þ (t, 3H, J¼6.9 Hz, CH3); m/z 442 [MþH] ; HR-MS calculated J¼8.7 Hz, ArH), 6.48 (s, 1H, ArH), 4.69 (s, 2H, COCH2), þ for C23H27N3O4S: m/z 442.1789 [MþH] , found 442.1795. 3.60 (t, 2H, J¼6.9 Hz, CH2), 2.12 (s, 3H, CH3), 1.42 1-[4-(2-Ethoxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(6-metho (m, 2H, CH2), 1.231.21 (m, 4H, CH22), 0.82 1 þ xylbenzo[d]thiazol-2-yl) urea (19k): H NMR (DMSO-d6, (t, 3H, J¼6.9 Hz, CH3); m/z 378 [MþH] ; HR-MS calcu- þ 300 MHz), d: 10.05 (s, 1H, CONH), 7.43 (s, 1H, ArH), lated for C18H23N3O4S: m/z 378.1496 [MþH] , found 7.33 (m, 1H, ArH), 7.21 (d, 2H, J¼8.7 Hz, ArH), 6.90–6.98 378.1482. (m, 3H, ArH), 4.80 (s, 2H, COCH2), 4.15–4.22 (q, 2H, 1-[4-(2-Hydroxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(5-chlor- 1 J¼6.9 Hz, CH2), 3.76 (s, 3H, OCH3), 3.65 (t, 2H, othiazol-2-yl)urea (20g): H NMR (DMSO-d6, 300 MHz), d: J¼7.2 Hz, CH2), 1.44 (m, 2H, CH2), 1.16–1.25 (m, 10.35 (s, 1H, CONH), 7.32 (s, 1H, ArH), 7.20 (d, 2H, 7H, CH22, CH3), 0.83 (t, 3H, J¼6.9 Hz, CH3); m/z J¼8.7 Hz, ArH), 6.94 (d, 2H, J¼8.7 Hz, ArH), 4.69 (s, 2H, þ 472 [MþH] ; HR-MS calculated for C24H29N3O5S: m/z COCH2), 3.60 (t, 2H, J¼7.5 Hz, CH2), 1.42 (m, 2H, þ 472.1906 [MþH] , found 472.1901. CH2), 1.24–1.23 (m, 4H, CH22), 0.80–0.84 (t, 3H, Novel phenyl-urea derivatives as dual-target ligands that can activate both GK and PPARg 597

þ J¼6.9 Hz, CH3); m/z 398 [MþH] ; HR-MS calculated for References þ C17H20ClN3O4S: m/z 398.0927 [MþH] , found 398.0936. 1-[4-(2-Hydroxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(5-hydro- 1. Wild S, Roglic G, Green A, Sicree R, King H. Global prevalence 1 xylformylthiazol-2-yl) urea (20h): H NMR (DMSO-d6, of diabetes: estimates for the year 2000 and projections for 2030. 300 MHz), d: 12.84 (s, 2H, COOH 2), 10.38 (s, 1H, Diabetes Care 2004;27:1047–53. CONH), 7.82 (s, 1H, ArH), 7.20 (d, 2H, J¼8.7 Hz, 2. Saltiel AR, Kahn CR. Insulin signalling and the regulation of glucose and lipid metabolism. Nature 2001;414:799–806. ArH), 6.95 (d, 2H, J¼8.7 Hz, ArH), 4.69 (s, 2H, COCH2), 3. Zhang M, Chen L. Berberine in type 2 diabetes therapy: a new 3.63 (t, 2H, J¼7.2 Hz, CH2), 1.43 (m, 2H, CH2), 1.14– perspective for an old antidiarrheal drug?. Acta Pharm Sin B 1.28 (m, 4H, CH22), 0.80–0.84 (t, 3H, J¼6.9 Hz, CH ); m/z 408 [MþH]þ; HR-MS calculated for 2012;2:378–86. 3 4. Matschinsky FM, Glaser B, Magnuson MA. Pancreatic beta-cell C H N O S: m/z 408.1216 [MþH]þ, found 408.1224. 18 21 3 6 glucokinase: closing the gap between theoretical concepts and 1-[4-(2-Hydroxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3- 1 experimental realities. Diabetes 1998;47:307–15. (benzo[d]thiazol-2-yl)urea (20i): H NMR (DMSO-d6, 5. Grimsby J, Sarabu R, Corbett WL, Haynes NE, Bizzarro FT, 300 MHz), d: 7.78 (d, 1H, J¼7.5 Hz, ArH), 7.39–7.29 (m, Coffey JW, et al. Allosteric activators of glucokinase: potential 2H, ArH), 7.22–7.15 (m, 3H, ArH), 6.96 (d, 2H, J¼8.4 Hz, role in diabetes therapy. Science 2003;301:370–3. ArH), 4.80 (s, 2H, COCH2), 4.15–4.20 (q, 2H, J¼6.9 Hz, 6. Moller DE. New drug targets for type 2 diabetes and the CH2), 3.66 (t, 2H, J¼7.2 Hz, CH2), 1.45 (m, 2H, metabolic syndrome. Nature 2001;414:821–7. CH2), 1.24–1.25 (m, 4H, CH22 ), 0.83 (t, 3H, 7. Moller DE, Greene DA. Peroxisome proliferator-activated recep- þ J¼6.9 Hz, CH3); m/z 414 [MþH] ; HR-MS calculated for tor (PPAR) agonists for diabetes. Adv Protein Chem 2001;56:181– þ C21H23N3O4S: m/z 414.1486 [MþH] , found 414.1482. 212. 1-[4-(2-Hydroxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(6- 8. Ye JP. Challenges in drug discovery for thiazolidinedione sub- 1 stitute. Acta Pharm Sin B 2011;1:137–42. methylbenzo[d]thiazol-2-yl)urea (20j): H NMR (DMSO-d6, 300 MHz), d: 7.58 (s, 1H, ArH), 7.27 (s, 1H, ArH), 7.20 (d, 9. Ushiroda K, Maruta K, Takazawa T, Nagano T, Taiji M, Kohno 2H, J¼8.7 Hz, ArH), 7.12 (d, 1H, J¼7.8 Hz, ArH), 6.94 (d, T, et al. Synthesis and pharmacological evaluation of novel 2H, J¼8.7 Hz, ArH), 4.70 (s, 2H, COCH ), 3.65 (t, 2H, benzoylazole-based PPARa/g activators. Bioorg Med Chem Lett 2 2011;21:1978–82. J¼7.2 Hz, CH ), 2.34 (s, 3H, CH ), 1.45 (m, 2H, 2 3 10. Kumari V, Li C. Comparative docking assessment of glucokinase CH2), 1.24–1.29 (m, 4H, CH22), 0.83 (t, 3H, J¼6.9 þ interactions with its allosteric activators. Curr Chem Genomics Hz, CH3); m/z 428 [MþH] ; HR-MS calculated for þ 2008;2:76–89. C22H25N3O4S: m/z 428.1639 [MþH] , found 428.1639. 11. Bahn YS, Cox GM, Perfect JR, Heitman J. Carbonic anhydrase 1-[4-(2-Hydroxyl-2-oxoethoxy)phenyl]-1-(n-pentyl)-3-(6- and CO sensing during Cryptococcus neoformans growth, differ- 1 2 methoxylbenzo[d]thiazol-2-yl)urea (20k): H NMR (DMSO- entiation, and virulence. Curr Biol 2005;15:2013–20. d6,300MHz),d:7.42(s,1H,ArH),7.34(m,1H,ArH),7.21(d, 12. Ishikawa M, Nonoshita K, Ogino Y, Nagae Y, Tsukahara D, 2H, J¼8.7 Hz, ArH), 6.90–6.96 (m, 3H, ArH), 4.70 (s, 2H, Hosaka H, et al. Discovery of novel 2-(pyridine-2-yl)-1H-benzi- COCH2), 3.76 (s, 3H, OCH3), 3.65 (t, 2H, J¼7.2 Hz, midazole derivatives as potent glucokinase activators. Bioorg Med CH2), 1.44 (m, 2H, CH2), 1.24–1.25 (m, 4H, CH22), Chem Lett 2009;19:4450–4. þ 0.83 (t, 3H, J¼6.9 Hz, CH3); m/z 444 [MþH] ;HR-MS 13. Pfefferkorn JA, Lou J, Minich ML, Filipski KJ, He M, Zhou R, þ et al. Pyridones as glucokinase activators: identiﬁcation of a calculated for C22H25N3O5S: m/z 444.1593 [MþH] , found 444.1588. unique metabolic liability of the 4-sulfonyl-2-pyridone hetero- cycle. Bioorg Med Chem Lett 2009;12:3247–52. 14. Grimsby J, Berthel SJ, Sarabu R. Glucokinase activators for the Acknowledgments potential treatment of type 2 diabetes. Curr Top Med Chem 2008;8:1524–32. This project was ﬁnancially supported as National S&T Major 15. Kamata K, Mitsuya M, Nishimura T, Eiki J, Nagata Y. Special Project on Major New Drug Innovation (No. Structural basis for allosteric regulation of the monomeric 2009zx09103-036) and by the National Nature Science Foun- allosteric enzyme human glucokinase. Structure 2004;12:429–38. dation of China (No. 30572256).