INTRODUCTION:

SGLT-2 inhibitors are also called as gliflozins. SGLT-2 is a class of medicine which inhibits reabsorption of glucose in kidney and lower blood sugar level. They are also used in the treatment of type II diabetes mellitus (DM-2). SGLT-2 inhibits the sodium-glucose transport protein-2. The gliflozins are used to treat type 2 diabetes mellitus but are most often used as second or third line agents instead of first-line because there are other medications on the market that have much longer safety record and are less expensive than gliflozins. Gliflozins may be a good option for patients who are failing with metformin monotherapy, especially if reducing weight is part of the underlying treatment.

They are used in combination, for example metformin plus gliflozin and the triple therapy metformin, sulfonylurea and gliflozin.[1]

MECHANISM OF ACTION:

Sodium glucose co transporters (SGLTs) are newly available drug which are used in treatment of early and late type 2 diabetes. It blocks the glucose reabsorption in kidney and increase urinary glucose excretion. Glucose excreted and plasma levels drop down lead to development of all glycemic parameters. This mechanism of action is depend on blood glucose level as well as different actions of thiazolidinediones (mediated through GLUTs), is independent of the actions of insulin. Therefore, there is minimum potential for hypoglycemia, not risk of overstimulation or tiredness of beta cells. Because their mode of action relies upon normal renal glomerular-tubular function, SGLT-2 efficacy is reduced in persons with renal impairment. [2][3]

FIG:- SGLT-2 INHIBITORS AND ITS MECHANISM OF ACTION.

The drugs available in the SGLT2 inhibitors class are Forxiga (Dapagliflozin), Invokana (Canagliflozin) and Jardiance (Empagliflozin)[4]

EMPAGLIFLOZIN:

Empagliflozin approved in FDA in January 2014. Empagliflozin has molecular formula is C₂₃H₂₇ClO₇ and molecular weight is 450.912 gm/mol. Empagliflozin is a C-glycosyl compound consisting of a beta-glucosyl residue having a (4-chloro-3-{4-[(3S)-tetrahydrofuran-3-yloxy]benzyl}phenyl group at the anomeric centre. A sodium-glucose co-transporter 2 inhibitor used as an adjunct to diet and exercise to improve glycemic control in adults with type 2 diabetes mellitus. It has a role as a sodium-glucose transport protein subtype 2 inhibitor and a hypoglycemic agent. It is a C-glycosyl compound, an aromatic ether, a tetrahydrofuryl ether and a member of monochlorobenzenes. [5]

PHARMACOLOGICAL ACTIONS:

SGLT2-inhibitos function not only as antidiabetics for normalizing blood glucose, but also have diuretic and hypotensive actions and resolve salt-sensitive hypertension which plays an important role in diabetes. Therefore, SGLT-2 inhibitors strongly prevent heart and renal failure. [6][7][8]

PHARMACODYNAMICS:

URINARY GLUCOSE EXCRETION:

In type 2 diabetic patients, urinary glucose excretion raise without delay following a dose of empagliflozin and was maintained at the end of a 4-week treatment period averaging at approximately 64gm per day with 10 mg empagliflozin and 78g per day with 25mg empagliflozin once daily. Dose-proportional exposure of empagliflozin over 8 days resulted in urinary glucose excretion ranging from 77.9g with the 10mg dose to 89.8g with the 100mg dose.

URINARY VOLUME:

In a 5-day study, mean 24-hr urine volume increase from baseline was 341ml on Day 1 and 135ml on Day 5 of empagliflozin 25mg once daily treatment.[9]

CARDIAC ELECTROPHYSIOLOGY:

In a randomized, placebo-controlled, active-comparator and crossover study 30 healthy subjects were administered a single oral dose of empagliflozin 25mg, empagliflozin 200mg (8 times the maximum dose), moxifloxacin, and placebo. Like other SGLT-2 inhibitors, empagliflozin lowers serum uric acid levels probably through an effect on the urate transporter.[10][11]

PHARMACOKINETICS:

ABSORPTION:

The pharmacokinetics of empagliflozin has been characterized in healthy volunteers and patients with Diabetes mellitus type 2 (T2DM) and no clinically relevant differences were noted between the two populations. After oral administration, peak plasma concentrations of empagliflozin were reached at 1.5hr post-dose with 78% bioavailability. After that, plasma concentration decline in a biphasic method has quick distribution phase and relatively slow terminal phase. Steady state means plasma AUC and Cmax were 1870 nmol/L and 259nmol/L, in that order with 10mg empagliflozin once daily treatment and 4740nmol/L and 687nmol/L respectively with 25mg empagliflozin once daily treatment.[12]

DISTRIBUTION:

The apparent steady-state capacity of distribution was expected to 73.8 L base on the population pharmacokinetic analysis. Oral administration of an empagliflozin solution to healthy subjects, 86.2% plasma protein binding and partitioning of red blood cell is 36.8%.[13]

METABOLISM:

In the pre-clinical species, oxidative metabolism of empagliflozin was the primary metabolism pathway, but in humans empagliflozin is mainly metabolized by glucuronidation by the uridine 50-diphospho-glucuronosyltransferase (UGT) isoforms UGT1A3, UGT1A8, UGT1A9 and UGT2B7 to three main glucuronide conjugates (2-O-, 3-O-, and 6-O-glucuronide), which are each present in human plasma at <10% of total drug related exposure and are thought to be inactive.[14] In vitro studies suggested that the primary route of metabolism of empagliflozin in humans is glucuronidation by the uridine 50-diphospho-glucuronosyltransferases UGT2B7, UGT1A3, UGT1A8 and UGT1A9.[15]

ELIMINATION:

The mean terminal half-life ranged from 5.6 to 13.1 hrs in single rising dose studies and from 10.3 to 18.8 hrs in multiple-dose studies [16] In humans, both renal (approx 54%, about half as unchanged parent drug) and fecal (approx 41%, mostly unchanged parent drug) excretion are important elimination routes in contrast to most observations in the non-clinical species in which fecal elimination predominates.[17]

SIDE EFFECTS:

Frequent urination, burning and painful urination, cloudy urine and back pain. In women vaginal odor, white or yellowish vaginal discharge (may be lumpy or look like cottage cheese) or vaginal itching. In men redness, itching or swelling of the penis; rash on the penis; foul smelling discharge from the penis; or pain in the skin around the penis, feeling tired, weak, or uncomfortable; along with a fever and pain, tenderness, redness, and swelling of the genitals or the area between the genitals and the rectum, flu-like symptoms, dry mouth, nausea and vomiting, stomach pain, unusual fatigue or tiredness, difficulty breathing, breath that smells fruity, decreased consciousness or confusion.[18]

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

TABLE 1: Analytical method development and validation of Empagliflozin and its combinaton by HPLC method.

Sr.No	Research work	Stationary Phase	Mobile Phase	Detection wavelength	Flow rate (ml/min)	Retention time (min)	Ref no.
1.	Method development and validation of RP-HPLC method for the estimation of empagliflozin in API	Intersil C₁₈, (150mmx40mm, 5 μm)	Acetate buffer and methanol in the ratio of 30:70 v/v	260 nm	2 ml/min	1.223 min	19
2.	Method development and validation of empagliflozin by RP-HPLC in bulk and pharmaceutical dosage form	BDS column and PDA detector	Ortho phosphoric acid and Acetonitrile 70:30 v/v	233 nm	1 ml/min	3.23 min	20
3.	A new simple method development, validation and forced degradation studies of empagliflozin by using RP-HPLC	Waters ODS (C18) RP Column, 250 mm x 4.6 mm. 5µm.	Phosphate buffer: acetonitrile 45:55 v/v (pH-2.8)	228 nm	1 ml/min	3.887 min	21
4.	A validated isocratic chiral liquid chromatographic method for the enantio selective separation of empagliflozin on new immobilized amylose based stationary phase	Chiralpak IG was used having length 250mm, Internal diameter of 4.6mm and particle size of 5µ	n-Hexane: Ethanol (65:35 v/v)	224 nm	0.8 ml/min	15.37(S) min 17.75(R) min	22
5	Method development and validation of empagliflozin by RP-HPLC in bulk and pharmaceutical dosage form	Hypersil BDS 150mm x 4.6 mm, 5m	0.1% ortho phosphoric acid and acetonitrile (70:30 v/v)	233nm	1 ml/min	3.23 min	23
Empagliflozin with combination
1.	Analytical method development and validation of new stability-indicating reverse-phase high-performance liquid chromatography method for simultaneous estimation of metformin hydrochloride and empagliflozin in tablet dosage form	BDS column (250×4.6 mm, 5 m particle size)	Buffer and acetonitrile in the ratio of 50:50 v/v	210 nm	1 ml/min	2.588 min Met 3.679 min Empa	24
2.	Development and validation of stability indicating RP-HPLC method for the simultaneous estimation of metformin hydrochloride and empagliflozin in bulk and in a synthetic mixture	Kromosil 250 x 4.6 mm, 5µm.	0.1% Ortho phosphoric acid buffer and acetonitrile 45:55 v/v	233 nm	0.8 ml/min	2.277 min Met 3.421 min Empa	25
3.	Stability indicating RP-HPLC method development and validation for estimation of empagliflozin and metformin hcl	Inertsil ODS-2 (250 × 4.6 mm, 5µm)	Phosphate buffer pH 3.5 and acetonitrile in proportion of 50:50 v/v	227 nm	1 ml/min	2.635 min Met 4.388 min Empa	26
4.	Simultaneous estimation of empagliflozin and linagliptin by RP-HPLC method	C18 (250 mm x 4.6 mm, 5 µm)	Phosphate buffer solution and methanol 70:30 v/v	240 nm	1 ml/min	3.026 min Empa 3.979 min Lina	27
5.	Stability indicating method development and validation for simultaneous estimation of linagliptin and empagliflozin in tablets by HPLC	Hypersil ODS 3V, 250 x 4.6 mm.5.0µ.	Mobile phase-A: buffer 100% mobile phase-B: water: acetonitrile (5:95 v/v) Preparation of diluent-1: mix acetonitrile and water in the ratio of 70:30 v/v. Preparation of diluent-2: mix acetonitrile and water in the ratio of 50:50 v/v.	225 nm	1 ml/min	5.435min Empa 8.345min Lina	28
6.	Validated RP-HPLC method for simultaneous determination of canagliflozin, dapagliflozin, empagliflozin and metformin	Reversed phase C18 column [250×4.6 mm-5µm	Acetonitrile and 0.05 M phosphate buffer pH 4 in a ratio 50:50 v/v	212 nm	1 ml/min	1.898 min Met 3.004 min Empa 3.560 min Dapa 4.414 min Cana	29
7.	LC–MS/MS determination of empagliflozin and metformin	C18 column (50 mm x 2.1mm, 1.7 µm)	0.1% aqueous formic acid :acetonitrile (75:25 v/v)	-	0.2 ml/min	-	30
8.	A new validated stability indicating RP-HPLC method for simutaneous estimation of metformin hydrochloride and empagliflozin in tablet dosage forms	Kromasil C18; 50 x 4.6 mm; 5m.	Acetonitrile and buffer [0.1% orthophosphoric acid (pH 2.8)] 50:50 v/v	260 nm	1 ml/min	2.192 min Met 3.200 min Empa	31
9.	RP-HPLC method development and validation for simultaneous determination of linagliptin and empagliflozine in tablet dosage form	ODS column (250 x 4.6mm, 5µ)	Buffer and acetonitrile (45:50 v/v)	245 nm	1 ml/min	2.2 min Lina 3.6 min Empa	32
10.	Stability indicating RP-HPLC method for simultaneous estimation of empagliflozin and linagliptin in bulk and pharmaceutical dosage forms	Agilent C18 column (4.6×150mm)5µ	Methanol : pH 3 phosphate buffer (70: 30 % v/v)	254 nm	1 ml/min,	2.365 min empa 3.907min lina	33
11.	Stability indicating simultaneous estimation of metformin and empagliflozin in pharmaceutical tablet dosage form by RP-HPLC	KromasilC18 column (250mm×4.6mm, 5µm particle size)	Buffer: Acetonitrile 45:55 v/v	226 nm	1.1 ml/min	2.182 min Met 2.908 min Empa	34
12.	Development and validation of stability indicating RP-HPLC method for simultaneous estimation of empagliflozine and linagliptin in tablet formulation	C18 column [BDS 250mm x 4.6 mm, 5µ]	0.1% Perchloric acid and Acetonitrile 60:40 v/v	230 nm	1 ml/min	2.05 min Empa 4.10 min lina	35

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

Sr.no	Research work	Stationary Phase	Mobile phase	Detection Wavelength	Flow rate (ml/min)	Retention time(min)	Ref. no
1.	A new RP-HPLC method development and validation of dapagliflozin in bulk and tablet dosage form	Waters C18, 5 µm particle size, 25 cm × 4.6 mm	Phosphate buffer and acetonitrile in the ratio of 60:40 v/v	237 nm	1 ml/ min	3.461 min	38
2.	Method development and validation of dapagliflozin in API by RP-HPLC and UV- Spectroscopy	BDS column	Acetonitrile and ortho phosphoric acid in the ratio of 55:4 v/v	203 nm	1.2ml/min	2.87 min	39
3.	RP-HPLC method for estimation of dapagliflozin from its tablet	Princeton C18 column	Acetonitrile: 0.1% triethylamine (pH-5.0) in the ratio of 50:50v/v	224 nm	1 ml/min	5.163 min	40
4.	Stability-Indicating RP-HPLC method development for simultaneous determination and estimation of dapagliflozin in raw and tablet formulation	C18 (150mm x 4.6mm, 5μm)	Methanol:water (75:25 v/v)	230 nm	1 ml/min	3.1 min	41
5.	Development and validation of dapagliflozin by reversed-phase high-performance liquid chromatography method and forced degradation studies	Hypersil BDS (250 mm × 4.6 mm, 5 μ)	Buffer:acetonitrile (60:40 v/v)	245 nm	1 ml/min	2.789 min	42
6.	Development and validation of a RP-HPLC method for the estimation of dapagliflozin In API	BDS Column (250×4.5mm, 5µ)	Ortho Phosphoric Acid And Acetonitrile (45:55 v/v)	245 nm	1 ml/min	2.963 min.	43
7.	Stability indicating RP-HPLC method for estimation of dapagliflozin in bulk and tablet dosage Form	Hypersil BDS C18 250 Mm X 4.6 Mm, 5 mM	0.1% Ortho phosphoric acid buffer and acetonitrile 50:50% v/v.	245 nm	1 ml/min	2.226 min	44
Dapagliflozin with combination
1.	A highly validated RP-HPLC method development for the simultaneous estimation of dapagliflozin and saxagliptin in tablet dosage forms	Standard BDS C8 column (50 ×4.6 mm, 5m)	Potassium dihydrogen phosphate: acetonitrile in the ratio 55:45 v/v	210 nm	1 .0ml/min	2.266min Dapa, 2.805min Saxa.	45
2.	Validated Rp-Hplc Method For Simultaneous Determination Of Canagliflozin, Dapagliflozin, Empagliflozin And Metformin	C18 column [250×4.6 mm-5µm p.s] Inertsil® ODS.	acetonitrile and 0.05 M potassium dihydrogen phosphate buffer PH 4 in a ratio [65:35, v/v]	212nm	1.0ml/min	1.898min Met 3.004min Empa 3.560minDapa 4.414min Cana	46
3.	Development Of A New Stability Indicating Rp-Hplc Method For Simultaneous Estimation Of Saxagliptine And Dapagliflozin And Its Validation As Per Ich Guidelines	XTerra C 18 column (150mm x 4.6mm x5µm particle size).	phoaphate buffer (pH 4) and Acetonitrile (50:50v/v)	225nm	1.0ml/min	2.1min Saxa, 2.8min Dapa	47
4.	A Validated Stability Indicating High-Performance Liquid Chromatographic Method For Simultaneous Determination Of Metformin Hcl And Dapagliflozin In Bulk Drug And Tablet Dosage Form	Hypersil BDS C18 (250 mm × 4.6 mm, 5 µ particle size)	buffer (0.1% orthophosphoric acid) adjusted to pH 6.8 with triethylamine:acetonitrile in the ratio of 50:50%	240 nm	1 .0ml/min	2.791min Met, 3.789min Dapa	48
5.	Development and Validation of Stability-Indicating RP-HPLC Method for Simultaneous Determination of Dapagliflozin and Saxagliptin in Fixed-Dose Combination	Xterra C-18 analytical column (150 mm × 4.6 mm , particle size 3.5 µ)	buffer and acetonitrile (53:47 v/v)	230 nm	1.2 ml/min	1.14min Dapa, 0.95min saxa	49

Canagliflozin:

INVOKANA is the brand name of canagliflozin available in the market. Canagliflozin is chemically known as [2S,3R,4R,5S,6R]-2-3- {5-(4-fluoro-phenyl)-thiophen-2-ylmethyl} 4-methyl-phenyl] -6- hydroxymethyl tetrahydro-pyran-3,4,5-triol . The empirical formula of canagliflozin is C₂₄H₂₅FO₅S with a molecular weight of 444.52g/mol. It is available in white to off white solid and melting point ranges from 95-105°C. It is soluble in many organic solvents like (methanol, Dimethyl sulfoxide) and insoluble in aqueous media. [50] Canagliflozin is a antidiabetic medication used to treatment of type 2 diabetes. It is less preferred than metformin. It is not recommended in type 1 diabetes. Canagliflozin is a sodium-glucose co-transporter 2 (SGLT-2) inhibitor. It works by increasing the amount of glucose lost in the urine. Elimination 1/2 life is 11.8 (10-13) hrs and bioavailability is 65%. [51]

This Table 3 of literature shows the simple, sensitive and rapid RP-HPLC method for the development and validation of canagliflozin single drug with the mobile phase, stationary phase, flow rate, wavelength and retention time.

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

Sr.no	Research work	Stationary phase	Mobile phase	Detection wavelength	Flow rate (ml/min)	Retention time(min)	Ref. no
1.	A novel validated RP-HPLC method for the estimation of canagliflozin in bulk and pharmaceutical dosage forms.	Inertsil ODS-3(250×4.6mm, 5µ)	combination of 0.02% Formic acid: Acetonitrile 40:60 v/v	230nm	1.2 ml/min	4.4 min	52
2.	Analytical method development and validation for the estimation of canagliflozin in bulk and formulation by RP- HPLC	Inertsil ODS-3 (250 × 4.6 mm, 5μ)	Ammonium acetate buffer (pH-4.5) and Acetonitrile in the ratio of 30:70 v/v	252 nm	1 ml/min	4.5 min	53
3.	A validated reverse phase liquid chromatographic method for the determination of canagliflozin	Phenomenex Gemini-NX C18 Column (250×4.6 mm, 5 μm particle size)	Acetonitrile: 1-octanesulphonic acid in a ratio of 70:30 v/v	245 nm	1 ml /min	3.5 min	54
4.	Development and validation of a stability-indicating reverse phase HPLC-PDA method for determination of canagliflozin in bulk and pharmaceutical dosage form	C18 (250×4.6 mm, 5 µm)	0.1% w/v ortho-phosphoric acid and acetonitrile in the ratio of 45:55 v/v	290 nm	1 ml/min	6.29 min	55
5.	development and validation of chromatographic method for estimation of canagliflozin in API and tablet dosage form	C18 column having 250 mm length, 4.6 mm internal diameter, 5μ particle size.	Methanol: Acetonitrile: 0.1 % Ammonium Acetate in the proportion of 40:40:20 v/v/v.	290 nm	1ml/min	4.1 min	56
Canagliflozin with combination
1.	Analytical Method Development and Validation of Canagliflozin and Metformin hcl by Using RP – HPLC	WATER’S (250x 4.6mm, 5um)	0.1% OPA: Methanol (60:40 v/v)	273 nm	0.5ml/min	2.693 min Met, 4.227min Cana.	57
2.	A validated stability indicating RP-HPLC method for simultanious determination of metformin and canagliflozin in pharmaceutical formulation	Kromosil C18 250 column	Phosphate buffer and acetonitrile in the ratio of 65:35 v/v	248 nm	1.0ml/min	2.413 min Met, 3.548 min Cana.	58
3.	Method Development and validation of simultaneous estimation of metformin and canagliflozin by using RP HPLC	Kromasil C18 column (250mm x 4.6mm x5µm particle size)	Acetonitrile:Buffer:Methanol (52:38:10 v/v/v)	254 nm	1.0ml/min	2.216 min Met, 3.223min Cana.	59

This Table 4 of literature shows formulation available in the empagliflozin drug, list of polymers, method used for formulation.

TABLE 4: Formulations available in Empagliflozin, Dapagliflozin and Canagliflozin drug.

Sr. No	Drug	Formulation Type	Polymer	Method Used	Ref .no
1.	Empagliflozin	Empagliflozin spherical Agglomerates	MCCPH112, Cassalpinina spinosa, Hpmck 100M, Sodium alginate, Magnesium sterate	Direct compression	60
2.	Dapagliflozin	Extended Release Tablet	Sodium carboxymethyl cellulose, HPMCK100M, Silica,Mag.sterate	Direct compression	61
3.	Canagliflozin	In-situ Gelling Liquid Oral Formulation	Sodium Alginate, Calcium carbonate	Gel	62
4.	Canagliflozin	Sustained Release Matrix Tablets	Eudragit RS 100 ,Xanthan gum , Carbopol , Lactose monohydrate , Microcrystalline cellulose , Talc, Magnesium stearate	Direct compression	63

CONCLUSION:

Literature presented in this article shows pharmacological parameters of all SGLT-2 inhibitor classes like empagliflozin, canagliflozin and dapagliflozin. Essentially these review emphases on method development and validation of empagliflozin, dapagliflozin and canagliflozin alone and with its combination presented in tabular form from table 1-3. Table 4 shows the formulation available in the SGLT-2 inhibitors. On the account of method development and validation as well as formulations, present literature is very useful for all researchers and currently working people on SGLT-2 inhibitors class.

ACKNOWLEDGEMENT:

Authors are thankful to the Department of Pharmacy, Sumandeep Vidyapeeth University, Piparia, Waghodia, Vadodara, Gujarat, India for providing all the facilities required for literature survey throughout the work.

CONFLICT OF INTEREST:

There are no conflicts of interest.

REFERENCES:

1. https://en.wikipedia.org/wiki/SGLT2_inhibitor.

2. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4269649.

3. https://www.google.com/searchq=mechanism+of+action+of+sglt2+inhibitorsandrlz=1C1ASVA_enIN785 IN785 and source=lnms and tbm=isch and sa=Xandved=0ahUKEwi66pbt IPjAhW1mOYKHbKSDK4Q_AUIECgBandbiw=1366andbih=625#imgrc=qHcGnfD7qk71XM.

4. https://medlineplus.gov/druginfo/meds/a614043.html.

5. https://pubchem.ncbi.nlm.nih.gov/compound/empagliflozin.

6. Kimura G. Diuretic action of sodium-glucose co transporter 2 inhibitors and its importance in the management of heart failure. Circ J 2016 25; 80(11): 2277-81.

7. Vallon V, Thomson SC. Targeting renal glucose reabsorption to treat hyperglycaemia. The pleiotropic effects of SGLT2 inhibition.Diabetologia. 2017;60(2): 215-25.

8. Chawla G, Chaudhary KK. A complete review of empagliflozin: Most specific and potent SGLT2 inhibitor used for the treatment of type 2 diabetes mellitus Diabetes and Metabolic Syndrome. Clin Res and Rev 2019;13:2001-08.

9. Heise T, Seman L, Macha S, Jones P, Marquart A, Pinnetti S. Safety, tolerability, pharmacokinetics and pharmacodynamics of multiple rising doses of empagliflozin in patients with type 2 diabetes mellitus. Diabetes Ther. 2013;4(2):331-45.

10. Pfeffer MA, Claggett B, Diaz R, Dickstein K, Gerstein HC, Køber LV. et al. Lixisenatide in patients with type 2 diabetes and acute coronary syndrome. N Engl J Med. 2015;373(23):2247-57.

11. Christoph W, John ML, Silvio EI, David F, Michaela M. Empagliflozin and clinical outcomes in patients with type 2 diabetes mellitus, established cardiovascular disease, and chronic kidney disease. Circulation. 2018;137:119-29.

12. Seman L, Macha S, Nehmiz G, Simons G, Ren B, Pinnetti S. Empagliflozin, a potent and selective SGLT2 inhibitor, induces dose-dependent glucosuria in healthy subjects. Clin. Pharmacol Drug Dev. 2013;2(2):152-61.

13. http://www.rxlist.com/glyxambi-drug/clinical-pharmacology.htm.

14. Taub ME, Ludwig-Schwellinger E, Ishiguro N, Kishimoto W, Yu H, Wagner K. Sex species and tissue-specific metabolism of empagliflozin in male mouse kidney forms an unstable hemiacetal metabolite that degrades to 4-Hydroxycrotonaldehyde, a reactive and cytotoxic species. Chem Res Toxicol. 2015;28(1):103-15.

15. Scheen AJ. Pharmacodynamics, efficacy and safety of sodiumeglucose co-transporter type 2 (SGLT2) inhibitors for the treatment of type 2 diabetes mellitus. Drugs. 2015;75(1):33-59.

16. Scheen AJ. Pharmacokinetic and pharmacodynamic profile of empagliflozin, a sodium glucose co-transporter 2 inhibitor. Clin Pharmaco 2014;53(3):213-25.

17. Tomlinson B, Hu M, Zhang Y, Chan P, Liu ZM. Evaluation of the pharmacokinetics, pharmacodynamics and clinical efficacy of empagliflozin for the treatment of type 2 diabetes. Expert Opin Drug Metabol Toxicol 2016 ;21:1-3.

18. https://medlineplus.gov/druginfo/meds/a614043.html.

19. Padmaja N, Veerabhadram G. Method development and validation of Rp-Hplc method for the estimation of empagliflozin in API. Int J Pharm Sci Res. 2016; 7(2): 724-27.

20. Soumika M, Sangeetha E, Mahender L .Method development and validation of empagliflozin by Rp-Hplc in bulk and pharmaceutical dosage Form . Int J Adv Pharma Sci 2016; 7(1): 3040-42.

21. Bolle SK, Illendula S, Rao KNV, Dutt HR. A new simple method development, validation and forced degradation studies of empagliflozin by using Rp-Hplc. Int J Pharm Bio Sci 2019; 9(1): 25-35.

22. Nakka M, Nakka S, Utpala BN. A validated isocratic chiral liquid chromatographic method for the enantio selective separation of empagliflozin on new immobilized amylose based stationary phase. Int J Inov Pharma Sci Res. 2017;10(5):81-92.

23. Shyamala, M.Soumika, E.Sangeetha, L.Mahender. Method development and validation of empagliflozin by Rp-Hplc In bulk and pharmaceutical dosage form. Int J Adv Pharm Sci 2016;7(1): 3040-42.

24. Geetha SA, Rajitha G, Yadav RY, Uma P. Analytical method development and validation of new stability-indicating reverse-phase high-performance liquid chromatography method for simultaneous estimation of metformin hydrochloride and empagliflozin in tablet dosage form. Asian J Pharm Clin Res 2019 ;12(1): 241-44.

25. C. Rupasi , M. Bhagavan. Development and validation of stability indicating Rp-Hplc method for the simultaneous estimation of metformin hydrochloride and empagliflozin in bulk and in a synthetic mixture. Int J Pharm 2016; 6(4): 138-47.

26. Patel AM, Chaudhary AB. Stability indicating Rp-Hplc method development and validation for estimation of empagliflozin and metformin Hcl. World J Pharm Sci 6(9): 872-85.

27. Bakshi A, Mounika A, Bhutada S, Dr. M. Bhagvan. Simultaneous estimation of empagliflozin and linagliptin by RP-HPLC method. World J Pharm Sci 7(8) :1062-71.

28. Nagunath S, Subramanian NS, Reddy K. Stability indicating method development and validation for simultaneous estimation of linagliptin and empagliflozin in tablets by HPLC. Saudi J Med Pharm Sci 2018;4(8) : 884-96.

29. Ghadir AK, Ismail S, Mohammed SG, Mohammed AH. Validated RP-HPLC method for simultaneous determination of canagliflozin, dapagliflozin, empagliflozin and metformin, IJPCBS 2018; 8(1): 1-13.

30. Bassam M, Ayoub SM. LC–MS/MS determination of empagliflozin and metformin. J Chroma Sci 2017;55 (7):742–47.

31. Vinay KD, Rao NS. A new validated stability indicating Rp-Hplc method for simutaneous estimation of metformin hydrochloride and empagliflozin in tablet dosage forms. Int Res J Pharm Med Sci. 2018;1(5):16-22.

32. Madhusudhan P , Reddy MR , Devanna N. RPHPLC method development and validation for simultaneous determination of linagliptin and empagliflozine in tablet dosage form. Int Adv Res J Sci, Eng andTech. 2015;2(2) :95-99.

33. Rao MS, Anusha K, Syed M. stability indicating Rp-Hplc method for simultaneous estimation of empagliflozin and linagliptin in bulk and pharmaceutical dosage forms. J Syn Natu Chem 2017;2(1):1-6.

34. http://ajrconline.org/abstractview.aspx?PID=2017-10-6-11.

35. Naazneen S, Sridevi A . Development and validation of stability indicating RP-HPLC method for simultaneous estimation of empagliflozine and linagliptin in tablet formulation. Scholars Research Library Der Pharm Let 2016;8(17):57-65.

36. Suma BV ,Deveswaran R. An overview on analytical methods for dapagliflozin- an antidiabetic drug.Int J Pharm Sci Res. 2019;10(6): 2688-92.

37. https://www.google.com/search?q=dapagliflozinandrlz=1C1ASVA_enIN785IN785andoq=dapaandaqs=chrome.1.69i57j0l5.5290j0j8andsourceid=chromeandie=UTF-8.

38. Debata J, Kumar S, Jha SK, Khan A. A new Rp-Hplc method development and validation of dapagliflozin in bulk and tablet dosage form. Int J Drug Dev Res. 2017;9(2):48-51.

39. Sanagapati M, Dhanalakshmi K ,Reddy N, Sreenivasa S. Method development and validation of dapagliflozin in API by Rp-Hplc and UV-spectroscopy. Int J Pharm Sci Drug Res. 2014; 6(3): 250-52.

40. Mante GV, Hemke AT, Umekar MJ. RP-HPLC method for estimation of dapagliflozin from its tablet. Int J Chem Tech Res. 2018;11(01): 242-48.

41. Shakirbash S. Development and validation of dapagliflozin by reversed-phase high-performance liquid chromatography method and it forced degradation studies. Asian J Pharm Clin Res. 2017;10(11):101-05.

42. Sanagapati M, Dhanalakshmi K, Reddy N, Sreenivasa S. Development and validation of a RP-HPLC method for the estimation of dapagliflozin in API. Int J Pharm sci res 2014;5(12): 5394-97.

43. Jeyabaskaran M, Rambabu C, Lakshmi MS. Stability indicating Rp-Hplc method for estimation of dapagliflozin in bulk and tablet dosage form. ICJPIR 2015;2(4):69-78.

44. Padmaja BR, Sivagami B, Chandrasekar R, Babu MN. A highly validated Rp-Hplc method development for the simultaneous estimation of dapagliflozin and saxagliptin in tablet dosage forms. Int J Pharma Sci Drug Res 2018;10(5):372-78.

45. Ghadir AK, Ismail S, Mohammed SG, Mohammed AH. Validated Rp-Hplc method for simultaneous determination of canagliflozin, dapagliflozin, empagliflozin and metformin. Int J Pharma chem bio sci. 2018;8(1):1-13.

46. Kommineni V, Chowdary KPR, Prasad SV. Development of a new stability indicating RP-HPLC method for simultaneous estimation of saxagliptine and dapagliflozin and its validation as per Ich guidelines. Indo American J Pharma Sci. 2017;4(09):2920-32.

47. Mohammad Y, Gowri SD . A validated stability indicating high-performance liquid chromatographic method for simultaneous determination of metformin hcl and dapagliflozin in bulk drug and tablet dosage form. Asian J Pharm Clin Res 2015;8(3):320-26.

48. Singh N, Bansal P, Maithani M, Chauhan Y. Development and validation of stability-indicating RP-HPLC method for simultaneous determination of dapagliflozin and saxagliptin in fixed-dose combination. New J Chem:1-28.

49. Sanagapati M, Dhanalakshmi K, Nagarjuna RG, Sreenivasa S. Method development and validation of dapagliflozin in API by Rp-Hplc and UV-Spectroscopy. Int J Pharma Sci Drug Res 2014; 6(3):250-52.

50. Housheh S, Bachour V. Analysis of canagliflozin, review. World J Pharm Res;6(4):306-16.

51. https://www.google.com/search?q=canagliflozinandrlz=1C1ASVA_enIN785IN785andoq=canagandaqs=chrome.1.69i57j0l5.9410j0j7andsourceid =chromeandie=UTF-8.

52. Marella VL, Syed A, Prasanna L, Nalluri BN. A novel validated RP-HPLC method for the estimation of canagliflozin in bulk and pharmaceutical dosage forms. Int J Adv Pharm Ana 2017; 07(03):24-27.

53. Bhatt D, Thatavarthi RB. Analytical method development and validation for the estimation of canagliflozin in bulk and formulation by RP-HPLC. Int J Pharm Sci Drug Res 2018;10(3):139-43.

54. Sreenivasulua S, Raob MR, ChandraSekhar KB. A validated reverse phase liquid chromatographic method for the determination of canagliflozin, world J of Pharm Sci; 6(2) 1119-32.

55. Kaur I, Wakode S, Singh HP, Manachanda S. Development and validation of a stability-indicating reverse phase Hplc-Pda method for determination of canagliflozin in bulk and pharmaceutical dosage form. Pharmaceutical Methods 2016;7(1) 54-62.

56. Ladva BJ, Dobariya PV, Pancholi HD, Nayak BS, Jain S. Development and validation of chromatographic method for estimation of canagliflozin in API and tablet dosage form. Int J Recent Sci Res 2016;7(5):10976-79.

57. Bakshi A, Reddy NS, Bhutada S, Raju MB. Analytical method development and validation of canagliflozin and metformin Hcl by using Rp – Hplc. Int J Pharm Sci. 2018;12(4):122-30.

58. Gaware D, Patil RN, Harole M. A validated stability indicating Rp-Hplc method for simultanious determination of metformin and canagliflozin in pharmaceutical formulation. World J Pharm Sciences.2017;4(12):631-40.

59. Bangaruthalli J, Gowri DS, Renuka MN, Akhila P, Vanga D. Method development and validation of simultaneous estimation of metformin and canagliflozin by using Rp-Hplc. Int J Sci and Eng Res 2018;9(11)1309-19.

60. Kavuluru VS, Kothapallibonnath CS. Characterization and evaluation of empagliflozin spherical agglomerates by direct compression method. Int J Recent Sci Res 2018;9(2):24394-98.

61. https://patents.google.com/patent/WO2017114227A1/en.

62. Bhatta RG, Sadashivaiah R, Kotappa SBB. Formulation, optimization and evaluation of in-situ gelling liquid oral formulation of a novel antidiabetic drug: canagliflozin. Ind J Pharm Edu Res. 2019;53(2) 121-28.

63. Srinivas M, Gulle SB, Devilal J, Nanumala SK: Development and in vitro characterization of canagliflozin sustained release matrix tablets. 2016; vol. 6 (2): 258-280.

Received on 07.08.2019 Modified on 12.09.2019

Research J. Pharm. and Tech 2020; 13(7): 3472-3479.

DOI: 10.5958/0974-360X.2020.00616.2

LC–MS/MS determination of empagliflozin and metformin

Stability indicating simultaneous estimation of metformin and empagliflozin in pharmaceutical tablet dosage form by RP-HPLC

Development and validation of dapagliflozin by reversed-phase high-performance liquid chromatography method and forced degradation studies

Development and validation of a RP-HPLC method for the estimation of dapagliflozin In API

A highly validated RP-HPLC method development for the simultaneous estimation of dapagliflozin and saxagliptin in tablet dosage forms

Development and validation of a stability-indicating reverse phase HPLC-PDA method for determination of canagliflozin in bulk and pharmaceutical dosage form

55. Kaur I, Wakode S, Singh HP, Manachanda S. Development and validation of a stability-indicating reverse phase Hplc-Pda method for determination of canagliflozin in bulk and pharmaceutical dosage form. Pharmaceutical Methods 2016;7(1) 54-62.