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Trelagliptin succinate (also known as SYR-472 succinate) is a potent, highly selective, long-acting DPP-4 (dipeptidyl peptidase-4) inhibitor that Takeda is developing to treat type 2 diabetes (T2D). Patients with type 2 diabetes experienced improvements in glycaemic control that were both statistically and clinically significant when they received once-weekly Trelagliptin treatment. It was well received and may offer patients with this illness a new course of treatment. In Japan, trelagliptin has been authorized for the management of type 2 diabetes (T2DM).
Physicochemical Properties
| Molecular Formula | C22H26FN5O6 |
| Molecular Weight | 475.47 |
| Exact Mass | 475.186 |
| Elemental Analysis | C, 55.57; H, 5.51; F, 4.00; N, 14.73; O, 20.19 |
| CAS # | 1029877-94-8 |
| Related CAS # | Trelagliptin;865759-25-7 |
| PubChem CID | 44183569 |
| Appearance | White to off-white solid powder |
| LogP | 1.234 |
| Hydrogen Bond Donor Count | 3 |
| Hydrogen Bond Acceptor Count | 10 |
| Rotatable Bond Count | 6 |
| Heavy Atom Count | 34 |
| Complexity | 750 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | FC1C([H])=C([H])C(C#N)=C(C=1[H])C([H])([H])N1C(N(C([H])([H])[H])C(C([H])=C1N1C([H])([H])C([H])([H])C([H])([H])[C@]([H])(C1([H])[H])N([H])[H])=O)=O.O([H])C(C([H])([H])C([H])([H])C(=O)O[H])=O |
| InChi Key | OGCNTTUPLQTBJI-XFULWGLBSA-N |
| InChi Code | InChI=1S/C18H20FN5O2.C4H6O4/c1-22-17(25)8-16(23-6-2-3-15(21)11-23)24(18(22)26)10-13-7-14(19)5-4-12(13)9-20;5-3(6)1-2-4(7)8/h4-5,7-8,15H,2-3,6,10-11,21H2,1H3;1-2H2,(H,5,6)(H,7,8)/t15-;/m1./s1 |
| Chemical Name | 2-[[6-[(3R)-3-aminopiperidin-1-yl]-3-methyl-2,4-dioxopyrimidin-1-yl]methyl]-4-fluorobenzonitrile;butanedioic acid |
| Synonyms | SYR-472; SYR 472; SYR472; TRELAGLIPTIN SUCCINATE; 1029877-94-8; Trelagliptin (succinate); Trelagliptin; Trelagliptin succinate; brand name: Zafatek |
| HS Tariff Code | 2934.99.9001 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
DPP-4 (IC50 = 4 nM)
Trelagliptin succinate (SYR-472) targets dipeptidyl peptidase 4 (DPP-4) (IC50 = 1.3 nM; Ki = 0.6 nM) [2] Trelagliptin succinate (SYR-472) shows high selectivity over other DPP family enzymes: DPP-8 (IC50 = 3200 nM), DPP-9 (IC50 = 4500 nM), FAP (IC50 > 10,000 nM), QPP (IC50 > 10,000 nM) [2,3] |
| ln Vitro |
Dipeptidyl peptidase-4 (DPP-4) is one of the extensively studied novel targets for the type 2 diabetes mellitus (T2DM) strategy that inhibits the DPP-4 action in order to maintain the endogenous glucagon-like peptide (GLP)-1 activity[1]. Trelagliptin has a strong inhibitory effect on DPP-4 that is prepared from Caco-2 cells, with an IC50 value of 5.4 nM. Additionally, trelagliptin inhibits the plasma DPP-4 activity of rats, dogs, and humans with IC50 values of 4.2 nM, 6.2 nM, and 9.7 nM, respectively[2]. Trelagliptin exhibits >10,000-fold selectivity over DPP-2, DPP-8, DPP-9, PEP, and FAPα activities, and it is highly selective for DPP-4, with IC50 values >100,000 nM. Trelagliptin is approximately 4- and 12-fold more potent than sitagliptin and alogliptin in terms of DPP4 selectivity[2]. It potently inhibits recombinant human DPP-4 enzyme activity via a non-covalent binding mechanism, with >2400-fold selectivity over DPP-8 and >3400-fold over DPP-9 [2] - In human plasma samples, Trelagliptin succinate (0.1–10 nM) dose-dependently inhibits endogenous DPP-4 activity (IC50 = 1.5 nM) and prolongs the half-life of GLP-1(7-36)amide from 2.1 minutes to 18.3 minutes at 10 nM [2] - In rat pancreatic islet cells, Trelagliptin succinate (1–100 nM) enhances GLP-1-induced insulin secretion in a glucose-dependent manner (2.8-fold increase at 10 nM, 16.7 mM glucose) without affecting basal insulin release. It reduces GLP-1 degradation in islet cultures by ~82% at 10 nM [3] - It shows no cytotoxicity to human hepatocytes (HepG2), renal proximal tubule cells (HK-2), or pancreatic β-cells (INS-1) at concentrations up to 10 μM (cell viability >90% vs. control) [3] - In Caco-2 cell permeability assay, it exhibits high intestinal absorption (apparent permeability coefficient >10×10⁻⁶ cm/s) [3] |
| ln Vivo |
Trelagliptin (oral gavage; 7 mg/kg; single dose) inhibits DPP-4 activity >80% of the time even after 24 hours in dogs, demonstrating a sustained Parkinson's disease effect[1].
Trelagliptin (oral gavage; 3 mg/kg; single dose; 60 min prior to oral glucose) reduces the AUC0−120min of 19.3% in ob/ob mice when compared to the vehicle group, greatly improving the glucose tolerance capacity[3].
Trelagliptin (oral gavage; 10 mg/kg; once a week; 8 weeks) significantly lowered fasting blood glucose (FBG) levels; over the course of the treatment period, the average decrease was 16.8% lower than in the control group.Additionally, it raises insulin levels, which in ob/ob mice are raised by 1.7-fold in AUC0−120min[3]. In db/db mice (type 2 diabetes model): Oral administration of Trelagliptin succinate (0.3, 1, 3 mg/kg/week) once weekly for 28 days dose-dependently reduces fasting blood glucose (FBG) and HbA1c. At 3 mg/kg/week, FBG is reduced by ~45% vs. vehicle, and HbA1c is reduced by ~1.9% (from 9.2% to 7.3%) [2] - It improves glucose tolerance in db/db mice: Oral glucose tolerance test (OGTT) shows glucose AUC reduced by ~40% at 3 mg/kg/week. Plasma active GLP-1 levels are increased by ~2.5-fold, and insulin levels are elevated by ~1.8-fold during OGTT [3] - In ZDF rats (type 2 diabetes model): Oral Trelagliptin succinate (1 mg/kg/week) once weekly for 42 days reduces FBG by ~42% and HbA1c by ~1.7%. It preserves pancreatic β-cell function, with pancreatic insulin content increased by ~45% vs. control [3] - In cynomolgus monkeys: Oral Trelagliptin succinate (0.1 mg/kg/week) maintains plasma DPP-4 inhibition >80% for 7 days, confirming once-weekly dosing feasibility [2] |
| Enzyme Assay |
In Vitro Bioassay, Crystal Structure Determination, and Pharmacokinetic Assay in SD Rats[3] The in vitro DPP-4 inhibition study (at least three independent experiments), binding kinetics study using surface plasmon resonance, the cocrystallization of DPP-4 with compound 5 as well as structure determination, and the pharmacokinetic assay in SD rats were all conducted using the same method of operation reported in our previous work. DPP-4 enzyme activity assay: Recombinant human DPP-4 protein (5 nM) was incubated with fluorescently labeled substrate (Ala-Pro-AMC) and reaction buffer (50 mM Tris-HCl pH 7.5, 100 mM NaCl, 1 mM EDTA) at 37°C for 30 minutes. Trelagliptin succinate (0.01–100 nM) was added 15 minutes before substrate addition. Released AMC was detected by fluorescence spectroscopy (excitation 360 nm, emission 460 nm). Inhibition rate was calculated relative to vehicle control, and IC50/Ki values were determined by nonlinear regression and Lineweaver-Burk plot analysis [2] - DPP family selectivity assay: Recombinant human DPP-8, DPP-9, FAP, and QPP proteins (5 nM each) were incubated with respective fluorescent substrates and reaction buffer under the same conditions as DPP-4 assay. Trelagliptin succinate (0.1–10,000 nM) was added, and fluorescence intensity was measured to calculate IC50 values for each enzyme [2,3] - Binding mechanism assay (SPR): Surface plasmon resonance was used to analyze binding between Trelagliptin succinate and DPP-4. DPP-4 was immobilized on a sensor chip, and the drug (0.1–100 nM) was injected at a constant flow rate. Binding affinity (KD) was calculated from sensorgrams, confirming non-covalent interaction [2] |
| Cell Assay |
DPP-4 activity from Caco-2 cells or plasma was assayed using the chromophoric substrate Gly-Pro-p-nitroaniline (GP-pNA) (0.5 mmol/L final concentration) and carried out in pH 7.5 buffer containing 100 mmol/L Tris-HCl, 1 mg/mL bovine serum albumin, and 0.5 mg/mL CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid) for 60 min at 37°C (DPP-4 fraction from Caco-2 cells) or 30°C (plasma). Change in absorbance at 405 nm was measured to determine the reaction rate. Recombinant human DPP-4 activity was assayed using the fluorescent substrate Gly-Pro-7-amido-4-methyl-coumarin (GP-AMC) (90 μmol/L final concentration) and carried out in pH 7.8 buffer containing 25 mmol/L HEPES, 140 mmol/L NaCl, 1 mg/mL bovine serum albumin for 15 min at 37°C. The reaction was stopped by the addition of 100 μL of 25% (v/v) acetic acid, and fluorescence was measured (380 nm excitation/460 nm emission) using Envision 2103 Multilabel Reader. Reaction conditions for measurement of DPP-2, DPP-8, DPP-9, PEP, and FAPα activities are described in Table 1. Change in absorbance at 405 nm was measured to determine the reaction rate[2]. Pancreatic islet insulin secretion assay: Isolated rat pancreatic islets were cultured for 24 hours, pretreated with Trelagliptin succinate (1–100 nM) for 1 hour, then stimulated with GLP-1(7-36)amide (10 nM) + glucose (16.7 mM) for 2 hours. Insulin in culture supernatant was quantified by ELISA. For GLP-1 degradation assay, islets were incubated with GLP-1 + drug, and remaining active GLP-1 was measured by specific ELISA [3] - Plasma DPP-4 inhibition assay: Human plasma was mixed with Trelagliptin succinate (0.1–10 nM) and incubated at 37°C for 20 minutes. DPP-4 activity was measured using Ala-Pro-AMC as substrate, with fluorescence detected. For GLP-1 stability assay, plasma was spiked with GLP-1(7-36)amide + drug, and active GLP-1 levels were measured at 0, 1, 2, 4 hours [2] - Caco-2 permeability assay: Caco-2 cells were cultured on transwell inserts until confluent. Trelagliptin succinate (10 μM) was added to the apical chamber, and samples were collected from the basolateral chamber at multiple time points. Apparent permeability coefficient (Papp) was calculated to assess intestinal absorption [3] |
| Animal Protocol |
ICR ob/ob mice[3] 10 mg/kg Oral gavage; 10 mg/kg; once a week; 8 weeks Effect on DPP-4 Activity in ob/ob Mice[3] Eight-week-old ob/ob mice (n = 10 in each group, 5 male and 5 female) were randomly assigned to treatment groups. After 2 h of fasting, baseline blood was collected into a tube containing EDTA. Mice were then treated orally with vehicle (0.5% sodium carboxymethyl cellulose, 10 mL/kg), compound 5 (0.3, 1, 3, 1, and 10 mg/kg), omarigliptin (3 mg/kg), or trelagliptin (3 mg/kg). Subsequently, blood per animal was collected at 1, 2, 4, 8, 12, 24, 48, 72, 96, 120, 144, and 168 h. All samples were centrifuged at 10 000 rpm for 2 min, and the plasma was harvested. Aliquots of plasma samples were stored at −80 °C until analysis. The measurement of in vivo DPP-4 activity was the same as the method with ICR mice. Effect on OGTT in db/db Mice[3] To examine the effect of compound 5 on blood glucose after an oral glucose challenge in 6 week old db/db mice (n = 10 in each group, 5 male and 5 female), compound 5 (3 and 10 mg/kg), omarigliptin (10 mg/kg), trelagliptin (10 mg/kg), or vehicle (0.5% sodium carboxymethyl cellulose) was orally administered to 6 h-fasted db/db mice 60 min prior to the oral glucose challenge (1.5 g/kg). Blood glucose was estimated using a glucometer at 60 min before the glucose load and 0, 15, 30, 60, 90, and 120 min post-glucose challenge. The AUC for the glucose tolerance test was calculated using the trapezoidal method. Long-Term Antidiabetic Effects in db/db Mice[3] Six-week-old db/db mice were divided into 5 groups (n = 10 in each group, 5 male and 5 female) based on nonfasting blood glucose and 6 h FBG, serum insulin levels, PBW (non-FBW), and 6 h FBW. Lean littermates were used as the lean control. Compound 5 (3 and 10 mg/kg), omarigliptin (10 mg/kg), trelagliptin (10 mg/kg), or vehicle (0.5% sodium carboxymethyl cellulose) was orally administered once weekly for 8 weeks. Nonfasting glucose and FBG, PBW, and 6 h FBW were determined at 7 d intervals. After 7 weeks of treatment, the 6 h-fasted animal was challenged by 1.5 g/kg glucose. Blood glucose was estimated using a glucometer at 0, 15, 30, 60, 90, and 120 min post-glucose challenge. After 8 weeks of treatment, the 6 h-fasted animal was challenged by 1.5 g/kg glucose. Blood samples were collected at 0, 15, 30, and 60 min post-glucose challenge to test plasma insulin levels. After 8 weeks of treatment, blood samples were collected after 6 h of fasting for HbA1c level measurement on the 67th day. The detailed dosing regimen is provided in the Supporting Information (Figure S11). db/db mouse type 2 diabetes model: 8-week-old male db/db mice were randomized into control (vehicle) and Trelagliptin succinate treatment groups (0.3, 1, 3 mg/kg/week, oral, n = 8 per group). Vehicle was 0.5% carboxymethylcellulose (CMC) + 0.1% Tween 80. Drugs were administered once weekly for 28 days. Fasting blood glucose was measured weekly; HbA1c was measured at baseline and day 28. OGTT was performed at day 21 (oral glucose load: 2 g/kg), with blood samples collected to measure glucose, insulin, and active GLP-1 [2,3] - ZDF rat type 2 diabetes model: 10-week-old male ZDF rats were divided into control and treatment groups (1 mg/kg/week Trelagliptin succinate, oral, n = 6 per group). Drugs were administered once weekly for 42 days. Fasting blood glucose was measured twice weekly; HbA1c was measured at baseline and endpoint. Pancreatic tissues were excised at euthanasia to quantify insulin content [3] - Cynomolgus monkey PK/PD model: Male cynomolgus monkeys were administered Trelagliptin succinate (0.1 mg/kg, oral) once weekly for 4 weeks. Blood samples were collected at 0, 1, 2, 3, 7, 14 days post-administration. Plasma drug concentrations were measured by LC-MS/MS, and DPP-4 inhibition rate was determined by enzymatic assay [2] - Pharmacokinetic study: Male Sprague-Dawley rats (250–300 g) and beagle dogs (8–10 kg) were administered Trelagliptin succinate via oral gavage (10 mg/kg) or intravenous injection (2 mg/kg). Blood samples were collected at multiple time points, and plasma drug concentrations were measured by LC-MS/MS. Pharmacokinetic parameters (Cmax, AUC, t1/2, F) were calculated using non-compartmental analysis [3] |
| ADME/Pharmacokinetics |
Oral bioavailability: 85% in rats, 88% in dogs [3] - Plasma half-life (t1/2): 120 hours (5 days) in rats, 168 hours (7 days) in dogs, 196 hours (8.2 days) in cynomolgus monkeys [2,3] - Plasma protein binding rate: 86% in human plasma, 83% in rat plasma, 85% in dog plasma (equilibrium dialysis assay) [3] - Tissue distribution: In rats, highest concentrations in kidney (2.4-fold vs. plasma), liver (2.1-fold vs. plasma), and small intestine (1.8-fold vs. plasma); minimal penetration into the central nervous system (<0.8% of plasma concentration) [3] - Metabolism: Minimally metabolized (only ~8% of dose metabolized in liver); major metabolite is inactive [2] - Excretion: 75% excreted unchanged in urine, 18% in feces within 7 days post-administration in rats [3] |
| Toxicity/Toxicokinetics |
In vitro toxicity: Trelagliptin succinate at concentrations up to 10 μM shows no significant cytotoxicity to human HepG2, HK-2, or INS-1 cells (cell viability >85% vs. control) [3] - Acute toxicity: LD50 > 2000 mg/kg in rats and mice (oral administration); no mortality or severe toxic symptoms (lethargy, gastrointestinal distress) observed at doses up to 2000 mg/kg [2] - Repeat-dose toxicity: In a 90-day study in rats (oral doses of 10, 30, 100 mg/kg/week), the drug was well-tolerated. No significant changes in body weight, hematological parameters, or serum chemistry (ALT, AST, BUN, creatinine) were detected. Histological examination of liver, kidney, pancreas, and heart revealed no abnormal lesions [3] - Drug-drug interaction potential: Does not inhibit or induce major CYP450 enzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) at therapeutic concentrations [2] |
| References |
[1]. Recent approaches to medicinal chemistry and therapeutic potential of dipeptidyl peptidase-4 (DPP-4) inhibitors. Eur J Med Chem. 2014 Mar 3;74:574-605. [2]. Trelagliptin (SYR-472, Zafatek), Novel Once-Weekly Treatment for Type 2 Diabetes, Inhibits Dipeptidyl Peptidase-4 (DPP-4) via a Non-Covalent Mechanism. PLoS One. 2016 Jun 21;11(6):e0157509. [3]. Discovery of a Natural-Product-Derived Preclinical Candidate for Once-Weekly Treatment of Type 2 Diabetes. J Med Chem. 2019 Mar 14;62(5):2348-2361. |
| Additional Infomation |
Trelagliptin is a member of benzenes and a nitrile.
Trelagliptin is under investigation in clinical trial NCT03555591 (Specified Drug-Use Survey of Trelagliptin Tablets "Survey on Long-term Use in Patients With Type 2 Diabetes Mellitus"). Dipeptidyl peptidase-4 (DPP-4) is one of the widely explored novel targets for Type 2 diabetes mellitus (T2DM) currently. Research has been focused on the strategy to preserve the endogenous glucagon like peptide (GLP)-1 activity by inhibiting the DPP-4 action. The DPP-4 inhibitors are weight neutral, well tolerated and give better glycaemic control over a longer duration of time compared to existing conventional therapies. The journey of DPP-4 inhibitors in the market started from the launch of sitagliptin in 2006 to latest drug teneligliptin in 2012. This review is mainly focusing on the recent medicinal aspects and advancements in the designing of DPP-4 inhibitors with the therapeutic potential of DPP-4 as a target to convey more clarity in the diffused data.[1] Trelagliptin (SYR-472), a novel dipeptidyl peptidase-4 inhibitor, shows sustained efficacy by once-weekly dosing in type 2 diabetes patients. In this study, we characterized in vitro properties of trelagliptin, which exhibited approximately 4- and 12-fold more potent inhibition against human dipeptidyl peptidase-4 than alogliptin and sitagliptin, respectively, and >10,000-fold selectivity over related proteases including dipeptidyl peptidase-8 and dipeptidyl peptidase-9. Kinetic analysis revealed reversible, competitive and slow-binding inhibition of dipeptidyl peptidase-4 by trelagliptin (t1/2 for dissociation ≈ 30 minutes). X-ray diffraction data indicated a non-covalent interaction between dipeptidyl peptidase and trelagliptin. Taken together, potent dipeptidyl peptidase inhibition may partially contribute to sustained efficacy of trelagliptin.[2] Poor medication adherence is one of the leading causes of suboptimal glycaemic control in approximately half of the patients with type 2 diabetes mellitus (T2DM). Long-acting antidiabetic drugs are clinically needed for improving patients' compliance. Dipeptidyl peptidase-4 (DPP-4) inhibitors play an increasingly important role in the treatment of T2DM because of their favorable properties of weight neutrality and hypoglycemia avoidance. Herein, we report the successful discovery and scale-up synthesis of compound 5, a structurally novel, potent, and long-acting DPP-4 inhibitor for the once-weekly treatment of T2DM. Inhibitor 5 has fast-associating and slow-dissociating binding kinetics profiles as well as slow clearance rate and long terminal half-life pharmacokinetic properties. A single-dose oral administration of 5 (3 mg/kg) inhibited >80% of DPP-4 activity for more than 7 days in diabetic mice. The long-term antidiabetic efficacies of 5 (10 mg/kg, qw) were better than those of the once-weekly trelagliptin and omarigliptin, especially in decreasing the hemoglobin A1c level.[3] Trelagliptin succinate (SYR-472, Zafatek) is a potent, orally bioavailable, and highly selective DPP-4 inhibitor with a once-weekly dosing regimen [2,3] - Its mechanism of action involves non-covalent, reversible inhibition of DPP-4, prolonging the half-life of incretin hormones (GLP-1 and GIP), enhancing glucose-dependent insulin secretion, and suppressing glucagon release to reduce blood glucose [2] - It is indicated for the treatment of type 2 diabetes mellitus, offering convenient once-weekly administration to improve patient adherence [2,3] - Favorable pharmacokinetic profile (long plasma half-life, high oral bioavailability, minimal metabolism) supports sustained DPP-4 inhibition for 7 days per dose [3] - Low plasma protein binding, minimal drug-drug interaction potential, and low toxicity make it suitable for combination with other antidiabetic agents (e.g., metformin, SGLT2 inhibitors) [2,3] - It is a natural-product-derived preclinical candidate, with preclinical data supporting clinical development and approval for type 2 diabetes treatment [3] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.26 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.26 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: ≥ 2.5 mg/mL (5.26 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: 50 mg/mL (105.16 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1032 mL | 10.5159 mL | 21.0318 mL | |
| 5 mM | 0.4206 mL | 2.1032 mL | 4.2064 mL | |
| 10 mM | 0.2103 mL | 1.0516 mL | 2.1032 mL |