A Review on HPLC Method Development and Validation for Gliptin Class: New Oral Antidiabetic Agents

 

Archana B. Gore, Manojkumar K. Munde*, Nikita B. Rukhe, Nilesh S. Kulkarni

PES Modern College of Pharmacy (for Ladies), Affiliated to Savitribai Phule Pune University, Moshi,

Pune-412105, Maharashtra, India.

 

ABSTRACT:

Gliptin is the class of antidiabetic medicine also called as dipeptidylpeptidase-4. DPP-4 (dipeptidyl peptidase-4) inhibitors (or "gliptins") represent a class of oral anti-hyperglycaemic agents that inhibit the enzyme DPP-4, thus augmenting the biological activity of the "incretin" hormones (glucagon-like peptide-1 [GLP-1] and glucose-dependent insulinotropic polypeptide [GIP]) Sitagliptin, Saxagliptin, Alogliptin, Linagliptin, Vildagliptin are the Gliptin class inhibitor for the treatment of type 2 diabetes mellitus and they decrease the breakdown of the incretin hormones such as glucagon like peptide 1 (GLP-1). All together gliptins have a good oral bioavailability which is not significantly influenced by food intake. PK/pharmacodynamics characteristics, that is, sufficiently prolonged half-life and sustained DPP-4 enzyme inactivation, generally allow one single oral administration per day for the management of T2DM; the only exception is vildagliptin for which a twice-daily administration is recommended because of a shorter half-life DPP-4. This paper is an updated review, providing an analysis of both the similarities and differences between the various compounds known as gliptins, currently used in the clinic (sitagliptin, saxagliptin, alogliptin linagliptin and vildagliptin). This paper discusses the pharmacokinetic and pharmacodynamic characteristics of gliptins. In this review we complied analytical method development and determination of the Gliptin inhibitors. Table no.1, 2, 3, 4, 5, shows the analytical method development and validation of Sitagliptin, Saxagliptin, Alogliptin, Linagliptin, and Vildagliptin alone and with its combination by the HPCL method.

 

 

 

INTRODUCTION:

Gliptin is also called as dipeptidylpeptidase-4 (ddp-4) inhibitors. Dipeptidylpeptidase-4(dpp-4) inhibitors offer new options for the management of type 2 diabetes. Glucagon increases blood glucose levels, and dpp-4 inhibitors decrease glucagon and blood glucose levels.

 

The mechanism of dpp-4 inhibitors is to increase incretin levels (glp-1 and gip), which inhibit glucagon release, which in turn increases insulin secretion, reduce gastric emptying, and decreases blood glucose levels.¹ They work by blocking the action of dpp-4, an enzyme which destroys a group of gastrointestinal hormones called incretins. incretins help stimulate the production of insulin when it is needed (e.g. after eating) and decrease the production of glucagon by the liver when it is not needed (e.g. during digestion). They also slow down digestion and reduce appetite. So by protecting incretins from injury, dpp-4 inhibitors help control the blood glucose levels.² Gliptins are effective in lowering blood glucose levels and, because they can help decrease appetite, may be beneficial for people needing to lose weight. They work by increasing insulin and suppressing glucagon. ³

 

Mechanism of action:

Dpp-4 inhibitors work by blocking the action of dpp-4, an enzyme which destroys the hormone incretin. Incretins help the body produce more insulin only when it is needed and reduce the amount of glucose being produced by the liver when it is not needed. ⁴

 

Structure:

 

Fig. 1 Mechanism of action

 

Sitagliptin:

Sitagliptin was the first oral dipeptidylpeptidase-4 inhibitor approved by the FDA for the treatment of type 2 diabetics. Long-term clinical studies demonstrated that sitagliptin was effective in decreasing HbA_1c levels and fasting glucose levels and enhance postprandial glucose excursions. Sitagliptin is Chemically known as (2R)-4-Oxo-4[3-(trifluoromethyl)-5, 6-dihydro[1, 2, 4]triazolo[4, 3-a]pyrazin-7(8H)-yl]1-(2, 4, 5-trifluorophenyl]butan-2-amide.⁵

 

Pharmacological action:

sitagliptin is a dipeptidylpeptidase-4 (DPP-4) inhibitor that improves glycaemic control by preventing DPP-4 from inactivating the incretin hormones glucagon-like peptide-1 and glucose-dependent insulinotropic polypeptide, thus prolonging incretin activity in response to ingestion of supplement.⁶

 

Pharmacodynamics:

Patients having type 2 diabetes mellitus, administration of sitagliptin led to inhibition of DPP-4 enzyme activity for a 24-hour period. After an oral glucose load or a meal, this dipeptidylpeptidase-4 inhibition resulted in a 2to3 fold increase in circulating levels of active GLP-1 and GIP, reduce glucagon concentrations, and increased responsiveness of insulin release to glucose, resulting in higher C-peptide and insulin concentrations. The rise in insulin with the reduce in glucagon was associated with lower fasting glucose concentrations and decrease glucose excursion following an oral glucose load. Sitagliptin did not lower blood glucose or cause hypoglycaemia in the studies with healthy subjects.

 

Cardiac Electrophysiology:

In a randomized, placebo-controlled crossover study, 79 healthy subjects were administered a single oral dose of sitagliptin 100 mg, sitagliptin 800 mg (8 times the recommended dose), and placebo. At the recommended dose of 100 mg, there was no effect on the QTc interval obtained at the peak plasma concentration, or at any other time during the study. Following the 800 mg dose, the maximum increase in the placebo-corrected mean change in QTc from baseline was observed at 3 hours postdose and was 8.0 msec. This increase is not considered to be clinically significant. At the 800 mg dose, peak sitagliptin plasma concentrations were approximately 11 times higher than the peak concentrations following a 100-mg dose.

 

In patients with type 2 diabetes mellitus administered sitagliptin 100 mg (N=81) or sitagliptin 200 mg (N=63) daily, there were no meaningful changes in QTc interval based on ECG data obtained at the time of expected peak plasma concentration.

 

Pharmacokinetics:

The pharmacokinetics of sitagliptin has been extensively characterized in healthy volunteers and patients with type 2 diabetes mellitus. Following a single oral 100-mg dose to healthy volunteers, mean plasma AUC of sitagliptin was 8.52 Μm hr, Cmax was 950 nm, and apparent terminal half-life (t_1/2) was 12.4 hours. Plasma AUC of sitagliptin increased in a dose-proportional manner and increased approximately 14% following 100 mg doses at steady-state compared to the first dose. The intra-subject and inter-subject coefficients of variation for sitagliptin AUC were small (5.8% and 15.1%). In healthy volunteers and in patients with type 2 diabetes mellitus the pharmacokinetics of sitagliptin was generally similar.

 

Absorption:

After oral administration of a 100 mg dose to healthy volunteers, peak plasma concentrations of sitagliptin were reached occurring 1 to 4 hours postdose. The absolute bioavailability of sitagliptin is approximately 87%.

 

Effect of Food:

Coadministration of a high-fat meal with sitagliptin had no effect on the pharmacokinetics of sitagliptin.

 

Distribution:

The mean volume of distribution at steady state following a single 100-mg intravenous dose of sitagliptin to healthy volunteer is approximately 198 liters. The fraction of sitagliptin reversibly bound to plasma proteins is low (38%).

 

Metabolism:

Sitagliptin oral dose, approximately 16% of the radioactivity was excreted as metabolites of sitagliptin. Six metabolites were detected at trace levels and are not expected to contribute to the plasma DPP-4 inhibitory activity of sitagliptin. In vitro studies indicated that the primary enzyme responsible for the limited metabolism of sitagliptin was CYP3A4, with contribution from CYP2C8.

 

Elimination:

Approximately 79% of sitagliptin is excreted unchanged in the urine with metabolism being a minor pathway of elimination. The apparent terminal t_1/2 following a 100 mg oral dose of sitagliptin was approximately 12.4 hours and renal clearance was approximately 350 mL/min.

 

Excretion:

Following administration of an oral sitagliptin dose to healthy subjects, approximately 100% of the administered radioactivity was eliminated in feces (13%) or urine (87%) within one week of dosing.

 

Elimination of sitagliptin occurs primarily via renal excretion and involves active tubular secretion. Sitagliptin is a substrate for human organic anion transporter-3 (hOAT-3), which may be involved in the renal elimination of sitagliptin. The clinical relevance of hOAT-3 in sitagliptin transport has not been established. Sitagliptin is also a substrate of P-glycoprotein (P-gp), which may also be involved in mediating the renal elimination of sitagliptin. However, cyclosporine, a P-gp inhibitor, did not reduce the renal clearance of sitagliptin. [7]

 

Side effect:

The more common side effects that can occur with sitagliptin include: upset stomach, diarrhoea, stomach pain, upper respiratory infection, stuffy or runny nose and sore throat, headache.

 

Serious side effect:

Pancreatitis Symptoms can include: severe pain in your stomach that won’t go away, and that can be felt from your stomach through your back, vomiting

 

Low blood sugar Symptoms can include: intense hunger, nervousness, shakiness, sweating, chills, and clamminess, dizziness, fast heart rate, light-headedness’, sleepiness, confusion, blurred vision, headache, depression, irritability, crying spells, nightmares and crying out in your sleep.

 

Severe allergic reaction. Symptoms can include: skin rash, hives, swelling of your face, lips, tongue, and throat, trouble breathing or swallowing.

 

Kidney problems. Symptoms can include: swelling of your feet, ankles, or legs, drowsiness, tiredness, chestpain, nausea, shortness of breath, producing less urinethan usual.[8]

 

Sitagliptin:

Sitagliptinis available in market brand name of Januvia. It is an antidiabetic medication used to treat type 2 diabetes.[9] sitagliptin chemically known as(R)-3-Amino-1-[3-(trifluormetyl)-5, 6, 7, 8-tetrahydro[1, 2, 4]triazol[4, 3-a]pyrazin-7-yl]-4-(2, 4, 5-trifluorphenyl)butan-1-on. It has Molecular formula C16 H15F6N5O and molecular weight 407gm/mol.[10] Sitagliptin is a white to off-powder and exhibits pH dependent aqueous solubility. It is soluble in water and N, N-dimethyl formamide, slightly soluble in methanol, soluble in ethanol, acetonitrile and acetone.[11] Dipeptidyl Peptidase 4 Inhibitor is mechanism of action of sitagliptin. Sitagliptinis a dipeptidyl peptidase-4 (DPP-4) inhibitor which is used in alliance with diet andexercise in the therapy of type 2 diabetes, either singleorin combination with other oral hypoglycemic agents.[12]

 

Fig.2 sitagliptin

 

This table 1 literature shows the simple sensitive and rapid RP-HPLC method for the development and validation of Sitagliptin and its combination with the mobile phase, stationary phase, flow rate, wavelength, and retention time.

 

TABLE 1. Analytical method development and validation of Sitagliptin and its combination by HPLC

 

Alogliptin-

Nesina is the brand name of Alogliptin available in the market. Alogliptin is chemically known as (R)-2-((6-(3-aminopiperidin-1-yl)-3-methyl-2, 4-dioxo-3, 4-dihydropyrimidin-1(2H)-yl)methyl)benzonitrile. The empirical formula of alogliptin is C₁₈H₂₁N₅O₂.

 

With molecular weight 339.4 g/mol.[19] The solubility of Alogliptin in eight pure solvents (water, methanol, ethanol, n-propanol, isopropanol, acetone, acetonitrile, N, N-dimethylformamide and has melting point 185-188°C.[20] alogliptin is an oral anti-diabetic drug in the DPP-4 inhibitor (gliptin) class. Alogliptin does not decrease the risk of heart attack and stroke. Like other members of the gliptin class, it causes little or no weight gain, exhibits relatively little risk of hypoglycaemia, and has relatively modest glucose-lowering activity.[21] Alogliptin inhibits dipeptidyl peptidase 4 (DPP-4), which normally degrades the incretins glucose-dependent insulinotropic polypeptide (GIP) and glucagon like peptide 1 (GLP-1). The inhibition of DPP-4 increases the amount of active plasma incretins which helps with glycemic control. [22]

 

Fig. 3 Alogliptin

 

This Table 2 literature shows the simple sensitive and rapid RP-HPLC method for the development and validation of Alogliptin and its combination with the mobile phase, stationary phase, flow rate, wavelength, and retention time.

 

TABLE 2. Analytical method development and validation of Alogliptin and its combination by HPLC

 

Saxagliptin:

On July 31, 2009 the FDA approved Saxagliptin. Saxagliptin was licensed for use throughout the European Union by the European Medicines Agency on December 1, 2009. Saxagliptin, sold under the brand name Onglyza, is an oral anti-diabetic drug. It has molecular formula C₁₈H₂₅N₃O₂ and 315.417 g·mol⁻¹ as molecular weight. Saxagliptin is used as monotherapy or in combination with other drugs for the treatment of type 2 diabetic. Early development was solely by Bristol-Myers Squibb; in 2007.[28] It is White to yellow or light brown, non-hygroscopic, crystalline powder; lightly soluble in water at 24 °C; slightly soluble in ethyl acetate; soluble in methanol, ethanol, isopropyl alcohol; acetonitrile, acetone, and polyethylene glycol 400. DPP-4 enzyme activity inhibits saxagliptin for a 24-hour period. It also decreased glucagon concentrations and increased glucose -dependent insulin secretion from pancreatic beta cells.[29]

 

Fig. 4 Saxagliptin

 

This Table 3 literature shows the simple sensitive and rapid HPLC method for the development and validation of Saxagliptin and its combination with the mobile phase, stationary phase, flow rate, wavelength, and retention time.

 

TABLE 3.Analytical method development and validation of Saxagliptin and its combination by HPLC

 

Linagliptin:

Linagliptin sold under the brand name Tradjenta in market. Linagliptin is chemically known as8-[(3R)-3-aminopiperidin-1-yl]-7-but-2-ynyl-3-methyl-1-[(4-methylquinazolin-2-yl) methyl] purine-2, 6-dion. molecular formula of alogliptin C₂₅H₂₈N₈O₂ and 472.54 as molecular weight.[34] Linagliptin is a crystalline white to yellowish solid.[35] solubility value at 313.15 K was obtained inmethanol (3.02 × 10 −3) inpure solvents, followed byethanol (2.31 × 10 −3), acetonitrile (1.72 × 10 −3), n-propanol (1.25 × 10 −3 ) andisopropanol (8.74 × 10 −4 ).[36] Linagliptin is a dipeptidyl peptidase-4 inhibitor. It works by increasing the production of insulin and decreasing the production of glucagon by the pancreas [37]

 

Fig.5 Linagliptin

 

This Table 4 literature shows the simple sensitive and rapid RP-HPLCmethod for the development and validation of Linagliptin and its combination with the mobile phase, stationary phase, flow rate, wavelength, and retention time.

 

TABLE 4. Analytical method development and validation Linagliptin and its combination by HPLC

 

Vildagliptin-

Thebrand name ofVildagliptin is Galvus. Vildagliptin is chemically known as 1-[2-[(3-Hydroxy-adamantyl) amino]acetyl]pyrrolidine-2-carbonitrile. Molecular formula of vildagliptin C₁₇H₂₅N₃O₂ and 303.4 g/mol as molecular weight. It is an oral anti-hyperglycaemic agent (anti-diabetic drug) of the dipeptidyl peptidase-4 (DPP-4) inhibitor class of drugs. Vildagliptin inhibits the inactivation of GLP-1 and GIP by DPP-4, allowing GLP-1 and GIP to potentiate the secretion of insulin in the beta cells and suppress glucagon release by the alpha cells of the islets of Langerhans in the pancreas. Vildagliptin has been shown to decrease hyperglycemia in type 2 diabetes mellitus. [44] Vildagliptin is a white to slightly yellowish or slightly greyish crystalline powder with a melting point/range of approximately 150৹C. It is smoothly soluble in water. [45]Oral bioavailability of vildagliptin 85%. [46]

 

This Table 5 literature shows the simple sensitive and rapid RP-HPLC method for the development and validation of Vildagliptin and its combination with the mobile phase, stationary phase, flow rate, wavelength, and retention time.

 

Fig.6 Vildagliptin

 

Table 5.Analytical method development and validation of Vildagliptin and its combination by HPLC

 

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Received on 11.06.2021         Modified on 21.10.2021

Accepted on 28.12.2021   ©AandV Publications All Right Reserved