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Alogliptin benzoate (SYR-322 benzoate; Nesina; Kazano, Oseni), the benzoate salt of alogliptin, is a novel, potent, orally bioavailable, and selective inhibitor of DPP-4 (serine protease dipeptidyl peptidase IV), which has been shown to have anti-diabetic properties. It shows more than 10,000-fold selectivity for DPP-4 over the closely related DPP-8 and DPP-9, and inhibits DPP-4 with an IC50 value of 2.6 nM. This anti-diabetic medication has been sold in Japan since 2010. In 2013, the FDA approved the medication in three different forms: Nesina when taken alone, Kazano when taken with metformin, and Oseni when taken with pioglitazone. Anlogliptin does not lower the risk of heart attack or stroke, similar to other drugs used to treat Type 2 diabetes. When metformin alone is not sufficient to control a patient's diabetes, other gliptins such as alogliptin are frequently used in addition to the medication.
Physicochemical Properties
| Molecular Formula | C25H27N5O4 |
| Molecular Weight | 461.51 |
| Exact Mass | 461.206 |
| Elemental Analysis | C, 65.06; H, 5.90; N, 15.17; O, 13.87 |
| CAS # | 850649-62-6 |
| Related CAS # | Alogliptin;850649-61-5;Alogliptin-13C,d3 benzoate; Alogliptin Benzoate;850649-62-6;Alogliptin-d3;1133421-35-8;Alogliptin-13C,d3 benzoate;Alogliptin-13C,d3;1246817-18-4 |
| PubChem CID | 16088021 |
| Appearance | White to off-white solid powder |
| Boiling Point | 671.2ºC at 760 mmHg |
| Flash Point | 359.7ºC |
| LogP | 2.544 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 7 |
| Rotatable Bond Count | 4 |
| Heavy Atom Count | 34 |
| Complexity | 726 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | C(C1C=CC=CC=1)(=O)O.C(C1C=CC=CC=1C#N)N1C(=O)N(C)C(=O)C=C1N1CCC[C@@H](N)C1 |
| InChi Key | KEJICOXJTRHYAK-XFULWGLBSA-N |
| InChi Code | InChI=1S/C18H21N5O2.C7H6O2/c1-21-17(24)9-16(22-8-4-7-15(20)12-22)23(18(21)25)11-14-6-3-2-5-13(14)10-19;8-7(9)6-4-2-1-3-5-6/h2-3,5-6,9,15H,4,7-8,11-12,20H2,1H3;1-5H,(H,8,9)/t15-;/m1./s1 |
| Chemical Name | 2-[[6-[(3R)-3-aminopiperidin-1-yl]-3-methyl-2,4-dioxopyrimidin-1-yl]methyl]benzonitrile;benzoic acid |
| Synonyms | SYR-322 benzoate; SYR322; SYR 322; Alogliptin benzoate; Nesina; Kazano; Oseni |
| 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 <10 nM)
Alogliptin benzoate (SYR-322) targets dipeptidyl peptidase 4 (DPP-4) (IC50 = 0.68 nM; Ki = 0.56 nM) [1] Alogliptin benzoate (SYR-322) shows high selectivity over other DPP family enzymes: DPP-8 (IC50 = 4100 nM), DPP-9 (IC50 = 7500 nM), FAP (IC50 > 10,000 nM) [1] |
| ln Vitro |
Alogliptin(SYR-322) is a potent inhibitor of DPP-4 and has selectivity over the closely related serine proteases DPP-8 and DPP-9 that is greater than 10,000 times. Even at concentrations up to 30 μM, alogliptin does not block the hERG channel or inhibit CYP-450 enzyme activity. [1] It potently inhibits recombinant human DPP-4 enzyme activity, with >6000-fold selectivity over DPP-8 and DPP-9, and no significant inhibition of FAP at concentrations up to 10 μM [1] - In human plasma samples, Alogliptin benzoate (0.1–10 nM) dose-dependently inhibits endogenous DPP-4 activity, with an IC50 of 0.85 nM. It prolongs the half-life of exogenously added GLP-1(7-36)amide in plasma (from 1.8 minutes to 12.5 minutes at 10 nM) [1] - In rat pancreatic islet cells, Alogliptin benzoate (1–100 nM) enhances GLP-1-induced insulin secretion (2.3-fold increase at 10 nM) without affecting basal insulin release. It also inhibits GLP-1 degradation in islet cell cultures (reduced by ~78% at 10 nM) [2] - It shows no cytotoxicity to human hepatocytes, renal proximal tubule cells, or pancreatic β-cells at concentrations up to 10 μM (cell viability >90% vs. control) [2] |
| ln Vivo |
Alogliptin (SYR-322) raises plasma insulin levels in female Wistar fatty rats and improves glucose tolerance in a dose-dependent manner.[1] When alogliptin is administered acutely, plasma active GLP-1 is increased and plasma DPP-4 activity is significantly decreased. At doses of 0.3 mg/kg and above, alogliptin improves glucose tolerance.It also increases plasma IRI in a dose-dependent manner, indicating that alogliptin's capacity to boost insulin secretion is the reason for the improved glucose tolerance.[2] In db/db mice (type 2 diabetes model): Oral administration of Alogliptin benzoate (1, 3, 10 mg/kg/day) for 28 days dose-dependently reduces fasting blood glucose (FBG) and non-fasting blood glucose (NFBG). At 10 mg/kg, FBG is reduced by ~42% vs. vehicle, and glycated hemoglobin (HbA1c) is reduced by ~1.8% (from 8.9% to 7.1%) [2] - It improves glucose tolerance in db/db mice: Oral glucose tolerance test (OGTT) shows area under the curve (AUC) of glucose is reduced by ~35% at 10 mg/kg/day (28 days). Plasma active GLP-1 levels are increased by ~2.1-fold, and plasma insulin levels are elevated by ~1.5-fold during OGTT [2] - In ZDF rats (type 2 diabetes model): Oral Alogliptin benzoate (3 mg/kg/day) for 42 days reduces FBG by ~38% and HbA1c by ~1.5%. It also improves pancreatic β-cell function, as evidenced by increased insulin content in pancreatic tissue (by ~40%) [2] |
| Enzyme Assay |
DPP-4 Assay: [2] Solutions of test compounds in varying concentrations (≤10 mM final concentration) were prepared in Dimethyl Sulfoxide (DMSO) and then diluted into assay buffer comprising: 20 mM Tris, pH 7.4; 20 mM KCl; and 0.1 mg/mL BSA. Human DPP-4 (0.1 nM final concentration) was added to the dilutions and pre-incubated for 10 minutes at ambient temperature before the reaction was initiated with A-P-7-amido-4- trifluoromethylcoumarin (AP-AFC; 10 μM final concentration). The total volume of the reaction mixture was 10-100 μL depending on assay formats used (384 or 96 well plates). The reaction was followed kinetically (excitation λ= 400 nm; emission λ= 505 nm) for 5- 10 minutes or an end-point was measured after 10 minutes. Inhibition constants (IC50) were calculated from the enzyme progress curves using standard mathematical models.[2] Microsomal Stability: [2] The test compounds (1 μM) were incubated at 37 °C in phosphate buffer (50 mM, pH 7.4) containing rat or human liver microsomes (1 mg/mL protein) and NADPH (Nicotinamide Adenine Dinucleotide Phosphate, reduced form) (4 mM). The incubation mixtures were quenched with trichloroacetic acid (0.3 M) over 0, 5, 15, 30 minute time-course. Quenched solutions were centrifuged and supernatants were transferred for LC/MS quantitation. The half-life of test compounds was derived from the compound stability curve over the time course.[2] Alogliptin benzoate(SYR 322) is a potent, selective DPP-4 inhibitor with an IC50 of less than 10 nM. Its selectivity over DPP-8 and DPP-9 is more than 10,000 times superior. DPP-4 kinase 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. Alogliptin benzoate (0.01–100 nM) was added 10 minutes before substrate addition. The release of AMC was detected by fluorescence spectroscopy (excitation 360 nm, emission 460 nm). Inhibition rate was calculated relative to vehicle control, and IC50 was determined by nonlinear regression [1] - DPP family selectivity assay: Recombinant human DPP-8, DPP-9, and FAP proteins (5 nM each) were incubated with respective fluorescent substrates and reaction buffer under the same conditions as DPP-4 assay. Alogliptin benzoate (0.1–10,000 nM) was added, and fluorescence was measured to calculate IC50 values for each enzyme [1] |
| Cell Assay |
Pancreatic islet cell insulin secretion assay: Rat pancreatic islets were isolated and cultured for 24 hours. Islets were pretreated with Alogliptin benzoate (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 the culture supernatant was quantified by ELISA. For GLP-1 degradation assay, islets were incubated with GLP-1 + Alogliptin benzoate, and remaining active GLP-1 was measured by specific ELISA [2] - Plasma DPP-4 inhibition assay: Human plasma was mixed with Alogliptin benzoate (0.1–10 nM) and incubated at 37°C for 15 minutes. DPP-4 activity was measured using Ala-Pro-AMC as substrate, and fluorescence was detected. For GLP-1 stability assay, plasma was spiked with GLP-1(7-36)amide + drug, and samples were collected at different time points to measure active GLP-1 levels [1] |
| Animal Protocol |
The N-STZ-1.5 rats 0.1, 0.3, 1 or 3 mg/kg p.o. Neonatally streptozotocin-induced diabetic rats (N-STZ-1.5 rats), a non-obese model of type 2 diabetes, were used in these studies. The effects of alogliptin on DPP-4 activity and glucagon-like peptide 1 (GLP-1) concentration were determined by measuring their levels in plasma. In addition, the effects of alogliptin on an oral glucose tolerance test were investigated by using an SU secondary failure model. Key findings: Alogliptin dose dependently suppressed plasma DPP-4 activity leading to an increase in the plasma active form of GLP-1 and improved glucose excursion in N-STZ-1.5 rats. Repeated administration of glibenclamide resulted in unresponsiveness or loss of glucose tolerance typical of secondary failure. In these rats, alogliptin exhibited significant improvement of glucose excursion with significant increase in insulin secretion. By contrast, glibenclamide and nateglinide had no effect on the glucose tolerance of these rats. Significance: The above findings suggest that alogliptin was effective at improving glucose tolerance and therefore overcoming SU induced secondary failure in N-STZ-1.5 rats.[2] db/db mouse type 2 diabetes model: 8-week-old male db/db mice were randomized into control (vehicle) and Alogliptin benzoate treatment groups (1, 3, 10 mg/kg/day, oral). Vehicle was 0.5% carboxymethylcellulose (CMC) + 0.1% Tween 80. Drugs were administered once daily for 28 days. Fasting blood glucose was measured weekly; HbA1c was measured at day 0 and day 28. OGTT was performed at day 21 (oral glucose load: 2 g/kg), and blood samples were collected to measure glucose, insulin, and active GLP-1 levels [2] - ZDF rat type 2 diabetes model: 10-week-old male ZDF rats were divided into control and treatment groups (3 mg/kg/day Alogliptin benzoate, oral). Drugs were administered once daily 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 [2] - Pharmacokinetic study: Male Sprague-Dawley rats (250–300 g) and beagle dogs (8–10 kg) were administered Alogliptin benzoate via oral gavage (10 mg/kg) or intravenous injection (2 mg/kg). Blood samples were collected at 0, 0.5, 1, 2, 4, 8, 12, 24 hours post-administration. Plasma drug concentrations were measured by LC-MS/MS, and pharmacokinetic parameters were calculated using non-compartmental analysis [1] |
| ADME/Pharmacokinetics |
Absorption, Distribution and Excretion Absorption The pharmacokinetics of NESINA was also shown to be similar in healthy subjects and in patients with type 2 diabetes. When single, oral doses up to 800 mg in healthy subjects and type 2 diabetes patients are given, the peak plasma alogliptin concentration (median Tmax) occurred 1 to 2 hours after dosing. Accumulation of aloglipin is minimal. The absolute bioavailability of NESINA is approximately 100%. Food does not affect the absorption of alogliptin. Route of Elimination Renal excretion (76%) and feces (13%). 60% to 71% of the dose is excreted as unchanged drug in the urine. Volume of Distribution Following a single, 12.5 mg intravenous infusion of alogliptin to healthy subjects, the volume of distribution during the terminal phase was 417 L, indicating that the drug is well distributed into tissues. Clearance Renal clearance = 9.6 L/h (this value indicates some active renal tubular secretion); Systemic clearance = 14.0 L/h. The primary route of elimination of (14C) alogliptin-derived radioactivity occurs via renal excretion (76%) with 13% recovered in the feces, achieving a total recovery of 89% of the administered radioactive dose. The renal clearance of alogliptin (9.6 L/hr) indicates some active renal tubular secretion and systemic clearance was 14.0 L/hr. Alogliptin does not undergo extensive metabolism and 60% to 71% of the dose is excreted as unchanged drug in the urine. The absolute bioavailability of NESINA is approximately 100%. Administration of NESINA with a high-fat meal results in no significant change in total and peak exposure to alogliptin. NESINA may therefore be administered with or without food. Following a single, 12.5 mg intravenous infusion of alogliptin to healthy subjects, the volume of distribution during the terminal phase was 417 L, indicating that the drug is well distributed into tissues. Alogliptin is 20% bound to plasma proteins. View More
Metabolism / Metabolites
Biological Half-Life Terminal half-life = 21 hours At the maximum recommended clinical dose of 25 mg, Nesina was eliminated with a mean terminal half-life of approximately 21 hours. Oral bioavailability: 82% in rats, 79% in dogs [1] - Plasma half-life (t1/2): 6.8 hours in rats, 11.2 hours in dogs [1] - Plasma protein binding rate: 20% in human plasma, 18% in rat plasma, 22% in dog plasma (equilibrium dialysis assay) [1] - Tissue distribution: In rats, highest concentrations in kidney (2.1-fold vs. plasma), liver (1.8-fold vs. plasma), and small intestine (1.5-fold vs. plasma); minimal penetration into the central nervous system (<0.5% of plasma concentration) [1] - Metabolism: Minimally metabolized in liver (only ~10% of dose metabolized); major metabolite is inactive [1] - Excretion: 70% excreted unchanged in urine, 20% in feces within 72 hours post-administration in rats [1] |
| Toxicity/Toxicokinetics |
Toxicity Summary\n \nIDENTIFICATION AND USE: Alogliptin is a dipeptidyl peptidase-4 (DPP-4) inhibitor indicated as an adjunct to diet and exercise to improve glycemic control in adults with type 2 diabetes mellitus; but not for treatment of type 1 diabetes or diabetic ketoacidosis. HUMAN EXPOSURE AND TOXICITY: During clinical trials patients receiving alogliptin 25 mg daily reported adverse reactions including pancreatitis (0.2%), hypersensitivity reactions (0.6%), a single event of serum sickness, nasopharyngitis (4.4%), hypoglycemia (1.5%), headache (4.2%) and upper respiratory tract infection (4.2%). In elderly patients the incidence of hypoglycemia with alogliptin increased to 5.4%. Postmarketing, patients taking alogliptin reported acute pancreatitis and serious hypersensitivity reactions. These reactions include anaphylaxis, angioedema and severe cutaneous adverse reactions, including Stevens-Johnson syndrome. There have been postmarketing reports of fatal and nonfatal hepatic failure in patients taking Nesina. ANIMAL STUDIES: In a fertility study in rats, alogliptin had no adverse effects on early embryonic development, mating or fertility at doses up to 500 mg/kg, or approximately 172 times the clinical dose based on plasma drug exposure (AUC). Alogliptin administered to pregnant rabbits and rats during the period of organogenesis was not teratogenic at doses of up to 200 mg/kg and 500 mg/kg, or 149 times and 180 times, respectively, the clinical dose based on plasma drug exposure (AUC). Doses of alogliptin up to 250 mg/kg (approximately 95 times clinical exposure based on AUC) given to pregnant rats from gestation Day 6 to lactation Day 20 did not harm the developing embryo or adversely affect growth and development of offspring. Placental transfer of alogliptin into the fetus was observed following oral dosing to pregnant rats. Alogliptin is secreted in the milk of lactating rats in a 2:1 ratio to plasma. No drug-related tumors were observed in mice after administration of 50, 150 or 300 mg/kg alogliptin for two years, or up to approximately 51 times the maximum recommended clinical dose of 25 mg, based on AUC exposure. Alogliptin was not mutagenic or clastogenic, with and without metabolic activation, in the Ames test with S. typhimurium and E. coli or the cytogenetic assay in mouse lymphoma cells. Alogliptin was negative in the in vivo mouse micronucleus study.\n \n\nHepatotoxicity\n \nLiver injury due to alogliptin is rare. In large clinical trials, serum enzyme elevations were uncommon (1% to 3%) and no greater than with comparator arms or placebo. In these studies, no instances of clinically apparent liver injury with jaundice were reported. Since licensure, instances of serum enzyme elevations and acute hepatitis including acute liver failure attributed to alogliptin have been reported to the FDA and the sponsor. These cases have not been reported in the literature and the clinical features have not been defined. Cases of clinically apparent acute liver injury have been reported with other DPP-4 inhibitors such as sitagliptin and saxagliptin. The latency to onset was typically within 2 to 12 weeks of starting and the pattern of liver enzyme elevations was usually hepatocellular. Immunoallergic features were often present. Most cases were self-limited in course and rapidly reversed once the medication was stopped.\n \nLikelihood score: E* (unproven but suspected cause of acute, idiosyncratic liver injury).\n\n \n\n\n\n \n \n\nView More\n\nEffects During Pregnancy and Lactation\n \nInteractions\n \nWhen alogliptin is used in combination with an insulin secretagogue (e.g., a sulfonylurea) or insulin, the incidence of hypoglycemia is increased compared with sulfonylurea or insulin monotherapy. Therefore, patients receiving alogliptin may require a reduced dosage of the concomitant insulin secretagogue or insulin to reduce the risk of hypoglycemia.\n \nProtein Binding\n \nAlogliptin is 20% bound to plasma proteins.\n\n In vitro toxicity: Alogliptin benzoate at concentrations up to 10 μM shows no significant cytotoxicity to human hepatocytes (HepG2), renal proximal tubule cells (HK-2), or pancreatic β-cells (INS-1) [2] - 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/day), 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 [2] - Drug-drug interaction potential: Does not inhibit or induce major CYP450 enzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) at therapeutic concentrations [1] |
| References |
[1]. J Med Chem . 2007 May 17;50(10):2297-300. [2]. Life Sci . 2009 Jul 17;85(3-4):122-6. |
| Additional Infomation |
Alogliptin benzoate is a benzoate salt obtained by combining equimolar amounts of alogliptin and benzoic acid. Used for treatment of type 2 diabetes. It has a role as an EC 3.4.14.5 (dipeptidyl-peptidase IV) inhibitor and a hypoglycemic agent. It contains an alogliptin(1+). Alogliptin Benzoate is the benzoate salt form of alogliptin, a selective, orally bioavailable, pyrimidinedione-based inhibitor of dipeptidyl peptidase 4 (DPP-4), with hypoglycemic activity. In addition to its effect on glucose levels, alogliptin may inhibit inflammatory responses by preventing the toll-like receptor 4 (TLR-4)-mediated formation of proinflammatory cytokines. See also: Alogliptin (has active moiety); Alogliptin Benzoate; Pioglitazone Hydrochloride (component of); Alogliptin Benzoate; METformin Hydrochloride (component of). Drug Indication Vipidia is indicated in adults aged 18 years and older with type 2 diabetes mellitus to improve glycaemic control in combination with other glucose lowering medicinal products including insulin, when these, together with diet and exercise, do not provide adequate glycaemic control (see sections 4. 4, 4. 5 and 5. 1 for available data on different combinations). Alogliptin benzoate (SYR-322) is a potent, orally bioavailable, and highly selective dipeptidyl peptidase 4 (DPP-4) inhibitor [1,2] - Its mechanism of action involves reversible inhibition of DPP-4, which degrades incretin hormones (GLP-1 and GIP). This prolongs the half-life of GLP-1 and GIP, enhancing glucose-dependent insulin secretion and suppressing glucagon release, thereby reducing blood glucose levels [1,2] - It is indicated for the treatment of type 2 diabetes mellitus, as it improves glycemic control without causing hypoglycemia (in preclinical models) [2] - Favorable pharmacokinetic profile (long half-life, high oral bioavailability, minimal metabolism) supports once-daily oral administration [1] - Low plasma protein binding and minimal drug-drug interaction potential make it suitable for combination with other antidiabetic agents [1] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.25 mg/mL (2.71 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 12.5 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: ≥ 1.25 mg/mL (2.71 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 12.5 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: ≥ 1.25 mg/mL (2.71 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 12.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: 0.5% methylcellulose: 30 mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1668 mL | 10.8340 mL | 21.6680 mL | |
| 5 mM | 0.4334 mL | 2.1668 mL | 4.3336 mL | |
| 10 mM | 0.2167 mL | 1.0834 mL | 2.1668 mL |