Yuanwei Chen

Design, Synthesis, and Biological Evaluation of Deuterated C-Aryl Glycoside as a Potent and Long-Acting Renal Sodium-Dependent Glucose Cotransporter 2 Inhibitor for the Treatment of Type 2 Diabetes

SGLT2 inhibitors deuterated at sites susceptible to oxidative metabolism were found to have a slightly longer tmax and half-life (t1/2), dose-dependent increase in urinary glucose excretion (UGE) in rats, and slightly superior effects on UGE in dogs while retaining similar in vitro inhibitory activities against hSGLT2. In particular, deuterated compound 41 has the potential to be a robust long-acting antidiabetic agent.

EGT1442, a potent and selective SGLT2 inhibitor, attenuates blood glucose and HbA1c levels in db/db mice and prolongs the survival of stroke-prone rats

Sodium glucose co-transporter 2 (SGLT2) is a renal type III integral membrane protein that co-transports sodium and glucose from filtrate to epithelium in the proximal tubule. Human subjects with homozygous or compound heterozygous mutations in SLC5A2 exhibit glucosuria without hypoglycemia or other obvious morbidity, suggesting that blockade of SGLT2 has the potential to promote normalization of blood glucose without hypoglycemia in the setting of type 2 diabetes. This report presents the in vitro and in vivo pharmacological activities of EGT1442, a recently discovered SGLT2 inhibitor in the C-aryl glucoside class. The inhibitory effects of EGT1442 for human SGLT1 and SGLT2 were evaluated in an AMG uptake assay and the in vivo efficacy of treatment with EGT1442 was investigated in rats and dogs after a single dose and in db/db mice after chronic administration. The effect of EGT1442 on median survival of SHRSP rats was also evaluated. The IC(50) values for EGT1442 against human SGLT1 and SGLT2 are 5.6μM and 2nM, respectively. In normal rats and dogs a saturable urinary glucose excretion was produced with an ED(50) of 0.38 and 0.09mg/kg, respectively. Following chronic administration to db/db mice, EGT1442 dose-dependently reduced HbA(1c) and blood glucose concentration without affecting body mass or insulin level. Additionally, EGT1442 significantly prolonged the median survival of SHRSP rats. EGT1442 showed favorable properties both in vitro and in vivo and could be beneficial to the management of type 2 diabetic patients.

Exploration of O-spiroketal C-arylglucosides as novel and selective renal sodium-dependent glucose co-transporter 2 (SGLT2) inhibitors

A series of novel O-spiroketal C-arylglucosides have been prepared and evaluated in cell-based functional assays for activity against human sodium-dependent glucose co-transporters 1 and 2 (SGLT1 and 2). The core spiro[isobenzofuran-1,2-pyran] structure proved to be an effective scaffold for diversification and a number of compounds with single digit nanomolar potency and high selectivity have been synthesized. Compound 5a promoted glucosuria when administered in vivo in rats and produced a significant blood glucose reduction effect.

C-aryl glucosides substituted at the 4'-position as potent and selective renal sodium-dependent glucose co-transporter 2 (SGLT2) inhibitors for the treatment of type 2 diabetes.

A series of C-aryl glucosides with various substituents at the 4-position of the distal aryl ring have been synthesized and evaluated for inhibition of hSGLT1 and hSGLT2. Introduction of alkyl or alkoxy substituents at the 4-position was found to improve SGLT2 potency, whereas introduction of a hydrophilic group at this position was deleterious. Compounds with alkoxy-, cycloalkoxy- or cycloalkenyloxy-ethoxy scaffolds exhibited good inhibitory activity and high selectivity toward SGLT2. Selected compounds were investigated for in vivo efficacy.

O-Spiro C-aryl glucosides as novel sodium-dependent glucose co-transporter 2 (SGLT2) inhibitors

Two series of O-spiro C-aryl glucosides were synthesized and tested for inhibition of hSGLT1 and hSGLT2. 6-O-Spiro C-aryl glucosides exhibited potent in vitro hSGLT2 inhibitory activity but 2-O-spiro C-aryl glucosides showed no in vitro hSGLT2 inhibitory activity at a screening concentration of 1microM.

Conformationally Constrained Spiro C-Arylglucosides as Potent and Selective Renal Sodium-Dependent Glucose Co-transporter 2 (SGLT2) Inhibitors

Diabetes is a highly prevalent modern disease with over 246 million people afflicted worldwide in 2007. A failure of glycemic homeostasis secondary to nutritional imbalance is considered to be the principle explanation for the alarming and increasing incidence of type 2 diabetes mellitus (DM2) in both developed and developing countries. Although a large number of antihyperglycemic agents have been developed to treat the disease, 63 % of DM2 patients fail to achieve the target levels of glycosylated hemoglobin (HbA1C<7 %) recommended by the American Diabetes Association, 4] and consequently these individuals are at risk of developing complications, such as accelerated cardiovascular disease, diabetic nephropathy, retinopathy and ulceration. Recently, renewed emphasis on the development of safe oral antidiabetic agents with a favorable cardiovascular profile has highlighted the attractions of inhibition of renal glucose resorption as a therapeutic mechanism. Sodium glucose co-transporter 2 (SGLT2) is a 672-amino acid, high-capacity, low-affinity transporter expressed nearly exclusively in the S1 and S2 segments of the renal proximal tubule and believed to mediate the majority of renal glucose resorption from the glomerular filtrate. Because the etiology of type 2 diabetes mellitus (DM2) depends on a hypertrophic adipose reservoir, mechanisms that promote glucose disposal by urinary output are therapeutically attractive compared to mechanisms that promote increased glucose assimilation by adipocytes. Selective inhibitors of SGLT2 are expected to be safe because individuals homozygous or compound heterozygous for mutations in SLC5A2, the gene encoding SGLT2, exhibit no significant morbidities. In contrast, penetrant alleles leading to SGLT1 deficiency are the genetic cause of glucose– galactose malabsorption syndrome, which is associated with severe neonatal diarrhea and failure to thrive. In particular, the high selectivity could potentially reduce the gastrointestinal side effect. Hence inhibitors selective for SGLT2 over SGLT1 are attractive candidates for development. Following the initial disclosure of T-1095A, a selective and potent SGLT2 inhibitor designed based on the naturally occurring inhibitor phlorizin, by Tanabe Seiyaku Co., Ltd. (Osaka, Japan), multiple classes of SGLT2 inhibitors have been reported, including Oand C-glucosides. The most advanced inhibitors currently undergoing clinical development in phase III trials, dapagliflozin (1) 16] and canagliflozin, are C-arylglucosides. The studies described here were directed at identifying metabolically robust agents with high selectivity towards SGLT2. Information gained from modeling studies and analysis of the crystal structure of dapagliflozin (1) suggested the possibility of creating novel and conformationally constrained chemotypes with improved potency for SGLT2 by cyclizing the 1and 6’-positions of the glucose moiety and glucose-proximal phenyl ring (Figure 1). Preliminary studies showed that retention of a chlorine substituent at the 4’-position on the proximal phenyl ring is critical for activity. The synthesis and evaluation of three series of novel analogues, which have a different scaffold than previously reported inhibitors, are described here. The synthesis of spiro[isobenzofuran-1,2’-pyran] analogues 12 a–e was addressed first (Scheme 1). Persilylated gluconolactone 3 was prepared in 89 % yield by the slow addition of trimethylsilyl chloride (TMSCl) to commercially available gluconolactone 2 in the presence of N-methylmorpholine. 21] Benzoic acid 4 was subjected to bromination with N-bromosuccinimide (NBS) followed by esterification to yield aniline 6. Sandmeyer reaction and subsequent oxidation of 7 provided the key electron-deficient tetra-substituted benzene 8. Friedel–Crafts acylation of R substituted benzenes generated the benzophenone 9. Selective reduction of the resulting ketone with triethylsilane and further reduction of the methyl ester gave the corresponding benzyl alcohol 10. Protection of the primary hydroxy group with chloromethyl methyl ether produced bromide 11. Lithium–halogen exchange and subsequent coupling with lactone 3 gave a mixture of lactols, which were converted in situ to the desired spiro[isobenzofuran-1,2’-pyran] derivatives 12 a–e in 40 to 63 % yield after purification by preparative thin layer chromatography (TLC). The synthesis of spiro[indane-1,2’-pyran] glucosides 19 a–e was more challenging than that of O-spiroketal C-arylglucosides analogues (Scheme 2). Benzyl alcohol 13 was oxidized with Dess–Martin reagent and subsequently subjected to [a] B. Lv, B. Xu, Prof. Dr. Y. Chen Chengdu Institute of Organic Chemistry (CIOC) Chinese Academy of Sciences (CAS) No. 16, Southion 2, the first circle road, 610041 Chengdu (P.R. China) Fax: (+ 86) 28-8525-9387 E-mail : 51popo51@163.com chenyw@cioc.ac.cn [b] B. Lv, Dr. Y. Feng, Dr. J. Dong, Dr. M. Xu, B. Xu, W. Zhang, Dr. Z. Sheng, Prof. Dr. Y. Chen Egret Pharma (Shanghai) Company, Ltd. Halei Road 1118, 201203 Shanghai (P.R. China) [c] Dr. A. Welihinda, Prof. Dr. B. Seed Theracos Inc. , 550 Del Rey Avenue, Sunnyvale, CA 94805-3528 (USA) Supporting information for this article is available on the WWW under http://dx.doi.org/10.1002/cmdc.201000051.

ortho-Substituted C-aryl glucosides as highly potent and selective renal sodium-dependent glucose co-transporter 2 (SGLT2) inhibitors

A series of 2-substituted C-aryl glucosides have been synthesized and evaluated for inhibition of hSGLT1 and hSGLT2. Introduction of an appropriate ortho substituent at the proximal phenyl ring adjacent to the glycosidic bond was found to improve SGLT2 inhibitory activity and dramatically increase selectivity for hSGLT2 over hSGLT1. Selected compounds were investigated for in vivo efficacy.

Effect of N -methyl deuteration on metabolism and pharmacokinetics of enzalutamide

Background The replacement of hydrogen with deuterium invokes a kinetic isotope effect. Thus, this method is an attractive way to slow down the metabolic rate and modulate pharmacokinetics. Purpose Enzalutamide (ENT) acts as a competitive inhibitor of the androgen receptor and has been approved for the treatment of metastatic castration-resistant prostate cancer by the US Food and Drug Administration in 2012. To attenuate the N-demethylation pathway, hydrogen atoms of the N–CH3 moiety were replaced by the relatively stable isotope deuterium, which showed similar pharmacological activities but exhibited favorable pharmacokinetic properties. Methods We estimated in vitro and in vivo pharmacokinetic parameters for ENT and its deuterated analog (d3-ENT). For in vitro studies, intrinsic primary isotope effects (KH/KD) were determined by the ratio of intrinsic clearance (CLint) obtained for ENT and d3-ENT. The CLint values were obtained by the substrate depletion method. For in vivo studies, ENT and d3-ENT were orally given to male Sprague Dawley rats separately and simultaneously to assess the disposition and metabolism of them. We also investigated the main metabolic pathway of ENT by comparing the rate of oxidation and hydrolysis in vitro. Results The in vitro CLint (maximum velocity/Michaelis constant [Vmax/Km]) of d3-ENT in rat and human liver microsomes were 49.7% and 72.9% lower than those of the non-deuterated compound, corresponding to the KH/KD value of ~2. The maximum observed plasma concentration, Cmax, and area under the plasma concentration -time curve from time zero to the last measurable sampling time point (AUC0–t) were 35% and 102% higher than those of ENT when orally administered to rats (10 mg/kg). The exposure of the N-demethyl metabolite M2 was eightfold lower, whereas that of the amide hydrolysis metabolite M1 and other minor metabolites was unchanged. The observed hydrolysis rate of M2 was at least ten times higher than that of ENT and d3-ENT in rat plasma. Conclusion ENT was mainly metabolized through the “parent→M2→M1” pathway based on in vitro and in vivo elimination behavior. The observed in vitro deuterium isotope effect translated into increased exposure of the deuterated analog in rats. Once the carbon–hydrogen was replaced with carbon–deuterium (C–D) bonds, the major metabolic pathway was retarded because of the relatively stable C–D bonds. The systemic exposure to d3-ENT can increase in humans, so the dose requirements can be reduced appropriately.

Design, synthesis, and biological evaluation of deuterated apalutamide with improved pharmacokinetic profiles

A series of deuterated apalutamide were designed and prepared. Compared to its prototype compound 18, deuterated analogues 19 and 21 showed obviously higher plasma concentrations and better PK parameters after oral administration in mice. In rats, N-trideuteromethyl compound 19 displayed 1.8-fold peak concentration (Cmax), and nearly doubled its drug exposure in plasma (AUC0-∞) compared to compound 18. Unsurprisingly, compounds 18 and 19 had similar affinity for AR in vitro. In summary, the deuteration strategy could obviously improve PK parameters of apalutamide.

Design, Synthesis, and Biological Evaluation of Dimorpholine Substituted Thienopyrimidines as Potential Class I PI3K/mTOR Dual Inhibitors

Dysfunctional signaling of the PI3K/AKT/mTOR pathway in cancer and its crucial role in cell growth and survival have made it a much desired target for cancer therapeutics. A series of dimorpholine substituted thienopyrimidine derivatives had been prepared and evaluated in vitro and in vivo. Among them, compound 14o was identified as a dual Class I PI3K and mTOR kinase inhibitor, which had an approximately 8-fold improvement in mTOR inhibition relative to the class I PI3K inhibitor 1 (pictilisib, GDC-0941). Western blot analysis confirmed the 14o mechanistic modulation of the cellular PI3K/AKT/mTOR pathway through inhibiting phosphorylation of both AKT and S6 in human cancer cell lines. In addition, 14o demonstrated significant efficacy in SKOV-3 and U87MG tumor xenograft models without causing significant weight loss and toxicity.