CARDIAC GLYCOSIDES SEMINAR.pptx

Cardiac Glycosides

1. CARDIAC GLYCOSIDES

2. INTRODUCTION What are Glycosides? Glycosides may be defined as the organic compounds from plants or animal sources which on enzymatic or acid hydrolysis give one or more sugar moieties along with non sugar moiety. Cardiac Glycosides : Cardiac glycosides are an important class of naturally occurring drugs having cardiac inotropic property which has, both, toxic as well as beneficial effects on the heart.

3. SCROPHULARIACEAE Example Digitalis purpurea APOCYANACEAE Example Strophanthus hispidus LILIACEAE Example Urginea indica Sources of Cardiac Glycosides

4. General Properties of Cardiac Glycosides Amorphous powder Bitter taste Odourless Soluble in water and alcohols Insoluble in Organic solvents Highly toxic in nature **Increased number of sugars enhances water solubility .**

5. HISTORICAL USE Extracted from the skin of Poison Toad Used as poison to A ssassinate people and to make poisonous arrows. MODERN USE Extracted from Plant Glycosides To treat congestive heart failure. Treatment of atrial fibrillation and flutter.

6. GENERAL MEACHANSIM OF ACTION FOR CARDIAC GLYCOSIDES Direct inhibition of Na + /K + ‐ATPase Inability of myocytes to pump Na + leading to intracellular accumulation of Na + Indirect inhibition of Na + / Ca ++ exchanger, which increases intracellular Ca ++ More Ca ++ released by the sarcoplasmic reticulum, thereby making more Ca ++ available to bind to troponin-C Increased force and velocity of contraction Topic Features Describe here the topic of the section

7. CLASSIFICATION According to the type of Lactone Ring CARDENOLIDES BUFADIENOLIDES

8. Contains a five membered lactone ring with one double bond. The lactone ring is attached to steroidal nucleus through C-17 position. Present in a large no. in nature. Examples : Digitoxin, Digoxin CARDENOLIDE BUFADIENOLIDE Contains a six membered lactone ring with two double bonds. The lactone ring is attached to steroidal nucleus through 17 ᵝ position. Occurrence is fewer in nature. Examples : Bufalin, Scillaren

9. CHEMISTRY OF CARDIAC GLYCOSIDES

10. GLYCONE MOIETY 3-4 Sugar molecules attached to aglycone moiety. Sugar attached to 3 rd position through b linkage Various types of sugars such as L-Rhamnose, D-Glucose, D-Fructose, D- Digitoxose , D- Digitalose , D- Cymarose , D- Sarmentose , D- Thevatose , L- Vallarose etc. Glycone moiety does not potentiates the medicinal activity, but it increases the solubilization of aglycone part in the body and it is beneficial in the absorption as well as distribution. Sugars predominantly exist as B conformation. Besides Glucose and Rhamnose they usually contain De- oxysugars . Presence of -CH3CO group on sugar affects lipophilic and kinetics of entire glycosides  

11. AGLYCONE MOIETY The term “ genin ” is used for aglycone moiety without sugar. Steroidal nucleus is important for the activity. 3B-OH group is attached to the sugar moiety. 14B-OH group is important for activity. Additional -OH groups at C-5 , C-11, C-16 may be present. 17B-OH is attached to the unsaturated lactone. No. of -OH groups increases on aglycone, it makes onset of action faster as well as it enhances metabolism. Cis A/B, Trans B/C, Cis C/D are present in the structure. It makes nucleus very stable so more active. Presence and attachment to a specific position to a steroidal nucleus of lactone is essential for proper activity. Size and degree of unsaturation varies with source of glycoside. Aglycone moiety more important than Glycone moiety. Cyclopentanoperhydrophenthrene 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19

12. LACTONE MOIETY The presence of Unsaturated lactone increases activity of Cardiac glycosides, but if it is saturated then it will lose its activity. PYRONE Butyrolactone Cardenolide Bufadienolide Butyrolactone ring Pyrone ring It has a C-23 Steroidal nucleus It has a C-24 Steroidal nucleus Found in Leguminosae, Cruciferae, Euphorbiaceae Found in Liliaceae, Ranunculeasae Example : Digitalis, Strophanthus Example : Squill

13. General Chemical Tests

14. Chemical Tests for Cardiac Glycosides Drug 50% ethanol and 0.1 ml 1% solution of m- dinitrobenzene in ethanol + + Violet colour appears due to presence of active methylene group Raymond’s Test : 2-3 drops of 20% sodium hydroxide solution

15. Chemical Tests for Cardiac Glycosides Crude drug Heat with 0.1 -5% solution of xanthydrol in glacial acetic acid + + Red Colour is produced due to presence of 2-deoxy sugar Xanthydrol Test : 1% Hydrochloric acid

16. Chemical Tests for Cardiac Glycosides Glycoside Mixture of 1% ferric sulphate solution in 5% glacial acetic acid + + Reddish Brown colour appears at the junction and the upper liquid layer is Bluish-Green due to presence of deoxy sugar Keller-Killiani test : 1-2 drops of concentrated sulphuric acid **Upper layer turns Red if squill is present**

17. Chemical Tests for Cardiac Glycosides Glycoside Few drops of Pyridine + + Appearance of a Silver Mirror shows positive test Tollen’s test : (silver mirror test) Few ml of Tollen’s Reagent (solution of silver nitrate and ammonia)

18. Chemical Tests for Cardiac Glycosides Drug extracted with CHCl3 Few drops of acetic anhydride + + Appearance of Violet ring which turns Blue and finally Green due to presence of sterol Liebermann Test For Sterols : Concentrated Sulphuric acid (from side wall of test tube)

19. Chemical Tests for Cardiac Glycosides Piece of lamina or thick section of leaf + Yellow To Orange is seen Baljet Test : Sodium picrate reagent

20. Chemical Tests for Cardiac Glycosides Solution of Glycosides + Development of Blue or Violet colour that fades out in 1-2 hours shows presence of cardenolides Kedde Test : Kedde reagent (equal volumes of 2% 3,5 dinitrobenzoic acid in menthol and 7.5% aqueous solution of KOH)

21. Chemical Tests for Cardiac Glycosides Drug Few ml of pyridine and 2 drops of nitroprusside + + Deep Red colour is produced Legal Test : 2-3 drops of 20% sodium hydroxide solution

22. Chemical Tests for Cardiac Glycosides Solution of Glycosides + Appearance of Blue or Violet colour that shows presence of cardenolides and bufadienolides Antimony trichloride Test : Solution of antimony tetrachloride and trichloroacetic acid and heat

23. PLANTS CONTAINING CARDIAC GLYCOSIDES Digitalis purpurea Digitalis lanata Urginea maritima Urginea indica

24. Synonym : Purple foxglove, Lady’s glove, Finger flower Biological source : Dried leaves of Digitalis purpurea . Family : Scrophulariaceae Shape : Ovate lanceolate to broadly ovate Colour : Dark greyish green Odour : Odourless Taste : Bitter Size : 30cm long, 4cm wide Leaves : Hairy, slightly pubescent on both surfaces. Margin : Crenate or serrate or dentate Apex : Sub acute DIGITALIS PURPUREA MORPHOLOGY

25. GEOGRAPHICAL SOURCES DIGITALIS PURPUREA USA INDIA ENGLAND OTHER PARTS OF EUROPE IRAQ MEXICO JAPAN

26. 1. Biennial herb (a plant which blooms in its second year and then dies) 2. Good quality of drug is obtained especially from cultivated plant. 3. Flourish best in well drained loose soil preferably of siliceous origin with some slight shade 4. Plant growing in sunny situation possesses the active qualities of herb compare to shade by trees 5. Best grown when allowed to seed itself. 6. Seeds should be mixed with fine sand in order to ensure even distribution. 7. Before sowing soil is sterilized. 8. Sown in spring, the plant does not blossom till the following year. CULTIVATION OF DIGITALIS PURPUREA

27. 9. In dry season, sufficient water is supplied. 10. In 1st year, long stalk with rosette of leaves is produced. 11. True plant shows dull pink or magenta not pale colored or spotted externally. CULTIVATION OF DIGITALIS PURPUREA

28. COLLECTION OF DIGITALIS PURPUREA 1. September to November by hand Collection 2. Organic matter and discolored leaves are avoided. 3. After collection, leaves should be dried as soon as possible at 60°C. 4. By quick drying, green color of leaves is maintained. 5. Drying is carried out till moisture is Not More Than 5%. 6. Packed under-pressure in airtight container.

29. Microscopy Of Digitalis purpurea Dorsiventral leaf Plenty of simple covering & glandular trichome on both surfaces Covering trichome: uniseriate, 3-4 celled long, collapsed cells, acute apex & finely warty cuticle. Glandular trichome: short unicellular stalk, bicellular or rarely unicellular head Anomocytic (irregular celled) stomata Trichome-stomata more in lower surface Pericycle: parenchymatous above & collenchymatous below Calcium oxalate crystals are absent

30. Chemical constituents of Digitalis purpurea It contains 0.2-0.45% mixture of both primary and secondary cardiac glycosides. Purpurea glycosides A and B and Glucogitoloxin are primary glycosides. Primary glycosides are less absorbed and less stable than secondary glycosides such as digitoxin, gitoxin , gitaloxin . Digitalis contains about 30 Glycosides which include verodoxin, glucoverodexin and odoroside H. Additionally, it also contains 2 saponin glycosides – digitonin and gitonin. Digitoxigenin

31. Chemical Tests for Digitalis Boil 1g of finely powdered digitalis with 10ml 70% alcohol (2-3 mins) and filter F iltrate + 5ml water + 0.5ml strong solution of lead acetate. Shake well and sperate the filtrate Transfer to a test tube containing 2ml conc. H₂SO₄ Extractive is dissolved in glacial acetic acid, cooled + 2 drops of FeCl3 solution Reddish brown layer acquiring bluish-green colour after standing is observed due to presence of digitoxose Clear filtrate + equal vol. of CHCl ₃. Evaporate to yield extractive Keller-Kiliani Test for Digitoxose :

32. Chemical Tests for Digitalis Dissolve the Extract in pyridine S odium nitroprusside solution and make it alkaline + + Baljet Test : Pink or Red colour is produced Legal Test : Section of Digitalis Sodium picrate solution Yellow to Orange colour is produced

33. STANDARDS OF QUALITY Foreign organic matter : Not more than 2% Loss on drying : Not more than 5% w/w, by drying to constant weight at 105 °C   Acid – insoluble ash : Not more than 5%

34. Uses of Digitalis purpurea ADULTERANTS Digitalis leaves are adulterated with : The leaves of Verbascum Thapsus The leaves of Primula vulgaris The leaves of Symphatum officinale It is used in the treatment of congestive cardiac failure. It is used in the treatment of atrial fibrillation, atrial flutter, superventricular tachycardia. It has a good effect on kidney and results in good diuresis. Also used in case of epilepsy and acute mania. The major disadvantage of digitalis is that it has ‘cumulative effect’ so administration of drug should be under strict medical supervision. STORAGE Digitalis is stored in a well closed container in a cool place away from light. Leaves must not contain more than 5% of moisture because it results in loss of cardiac activity. The powder is stored in containers with dehydrating agents like silica gel to absorb excess moisture.

35. Synonym : Woolly foxglove, Grecian foxglove Biological source : Dried leaves of Digitalis lanata . Family : Scrophulariaceae Shape : Sessile, linear lanceolate Size : 30cm long, 4cm wide Margin : Entire Apex : Acuminate Trichomes : 10-14 celled non- glandular trichomes DIGITALIS LANATA MORPHOLOGY

36. GEOGRAPHICAL SOURCES DIGITALIS LANATA USA INDIA NETHERLANDS ECUADOR

37. 1. Biennial herb (a plant which blooms in its second year and then dies) 2. Perennial herb (perennial (Latin per, "through", annus, "year") is a plant that lives for more than two years) 3. Light (sandy), medium loamy and heavy clay soil 4. Prefers acid-alkaline-neutral soil 5. Can grow in semi-shade or no shade. 6. Dry or moist soil 7. Propagated by seeds 8. Seeds are sown in early spring in a cold frame 9. Germination in 2-4 weeks at 20° C CULTIVATION OF DIGITALIS LANATA

38. COLLECTION OF DIGITALIS LANATA When they are large enough to handle, seedlings are transplanted into individual pots and planted them out in the summer.

39. Microscopy Of Digitalis lanata Upper Epidermis : Single layered, covered with a thick cuticle, cells have wavy and thick walls; Stomata are also occasionally seen on the upper epidermis. Trichomes : two kinds of trichomes are seen in the upper epidermis, multicellular, uniseriate covering trichomes and glandular trichomes with unicellular stalk. Lower epidermis : Identical to upper epidermis but numerous stomata. Mesophyll : It is differentiated into palisade and spongy parenchyma. Vascular bundle is present towards the ventral surface (upper epidermis) of the midrib. Calcium oxalate crystals are absent

40. Chemical constituents of Digitalis lanata It contains 5 primary glycosides and in all about 70 glycosides. The primary glycosides are identified as Lantasoide A, B, C, D, and E. The aglycones – digoxigenin and diginatigenin are specific to Digitalis lanata .

41. Uses of Digitalis lanata Allied Species Digitalis lutea : (Straw Foxglove) This drug is as potent as Digitalis purpurea. It is supposed to have less irritation and is used as a common substitute for the official drug. Digitalis thapsi : (Spanish Foxglove) This drug is 1.25 to 3 times more potent than Digitalis purpurea . It shows 3-4 times more activity than Digitalis purpurea. It is the commercial source of digoxin, lanatoside C. It is used in the treatment of congestive heart failure and atrial fibrillation. Use should always be supervised by a qualified practitioner since in excess they cause nausea, vomiting, slow pulse, visual disturbance, anorexia and fainting.

42. Comparison between Digitalis purpurea and Digitalis lanata Digitalis Purpurea Digitalis lanata It has a tea-like odour and is bitter in taste It has tea-like odour Contains Purpurea Glycoside A and B along with Saponins, flavonoids and Anthraquinones. Contains Lanatoside A, B, C, D, E Sclerenchyma and calcium oxalate crystals are absent Calcium oxalate crystals are absent Petiole is winged Petiole is sessile Surface of leaf is pubescent (covered with fine soft hair) Surface of leaf shows cilia at lower third part. Numerous fragments of non-glandular hair Glandular hair are present

43. Synonym : White squill, scilla, Drimia maritima Biological source : Dried slices of bulbs of Urginea maritima Family : Liliaceace Colour : Translucent yellowish-white Odour : Slight and characteristic Taste : Bitter and acrid Size : Entire bulb is 12-15 cm in diameter and 18-20 cm in height. The slices are 3-6 cm in length and 4-8 mm in width. Shape : The slices are concavo- convex shaped EUROPEAN SQUILL MORPHOLOGY

44. GEOGRAPHICAL SOURCES EUROPEAN SQUILL MOROCCO GREECE SPAIN ITALY ALGERIA FRANCE

45. The plant prefers light (sandy) and medium (loamy) soils and requires well-drained soil . The plant prefers acid, neutral and basic (alkaline) soils. The plant can tolerate strong winds but not maritime exposure. Seeds are sown as soon as it is ripe in a greenhouse. The seeds are sown thinly so that the seedlings can be left in the pot for their first growing season. Once the plant becomes dormant the young bulbs are divided, placing two to three bulbs in each pot. CULTIVATION OF EUROPEAN SQUILL

46. COLLECTION OF EUROPEAN SQUILL Bulbs are dug out from the soil in the end of August and external scaly leaves and central portion are removed. The slices are dried completely in the sunlight or by heat of the stove. The drug is stored in airtight and especially in moisture proof containers.

47. Microscopy Of Squill Single layer of polygonal elongated epidermis is present which is covered with the cuticle. Mesophyll region consists of acicular calcium oxalate crystals, mucilage sheath, small round starch grains and vascular bundle . T.S of Squill

48. Chemical constituents of European Squill Squill contains cardiac glycosides of bufadienolide type. Scillaren-A is the most important (about of total glycosides) It also contains Scillaren-B   It contains Glucoscillaren-A and Proscillaridin -A in small amounts. Along with that it contains xanthoscillide , flavonoids, mucilage, calcium oxalate, sinistrin and a volatile substance which causes irritation

49. Red variety of squill contains : 1. Anthocyanin (red color) 2. Scilliroside (glycoside which is toxic to rat)

50. Comparison between Scillaren A and Scillaren B Nature Crystalline Amorphous Colour Colourless White/ Yellowish white Taste Bitter Very Bitter Odour Odourless Odourless Solubility Comparatively less water soluble More soluble in water, ethanol and methanol Activity Less active More active Stability Less stable More Stable SCILLAREN - A SCILLAREN - B PROPERTIES

51. Chemical Tests for European Squill Drug extracted with CHCl3 Few drops of acetic anhydride + + Appearance of Violet ring which turns Blue due to presence of sterol Liebermann Test For Sterols : Concentrated Sulphuric acid (from side wall of test tube)

52. Chemical tests for European Squill Mucilage + Corallin soda Yellow stains Red stains Mucilage + Iodine Mesophyll region of squill contains mucilage, calcium oxalate and xanthoscillide **Squill shows negative results for Baljet and Legal test**

53. Uses of European Squill ADULTERANTS The common adulterants of squill plant are the bulbs of different species of ledebouria . It is a potent cardiotonic without having any cumulative effect (unlike Digitalis ). It is mostly employed in small doses as an effective expectorant specially in chronic bronchitis. It causes vomiting in relatively higher doses It also has some diuretic properties squill glycosides possess high therapeutic index and rapid elimination.

54. Synonym : Jangli pyaz , sea onion, scilla, urginea Biological source : Dried slices of bulbs of Urginea indica kunth Family : Liliaceace Colour : Squill bulbs are yellowish to white Odour : Slight and characteristic Taste : Bitter and acrid Size : 30-60 mm in length and 3-8 mm broad. Shape : The slices are united in groups of 4-8 which are curved INDIAN SQUILL MORPHOLOGY

55. GEOGRAPHICAL SOURCE OF INDIAN SQILL KONKAN COAST SAURASHTRA LOWER HIMALAYAS

56. Though it is not been cultivated, it grows well at a temperature of 15–20°C and in sandy soil. The bulbs grow to full size within 5 years. The bulbs are collected after flowering, cut in to small slices and dried under sun. CULTIVATION AND COLLECTION OF INDIAN SQUILL

57. Chemical constituents of Indian Squill Indian squill contains about 0.3% of cardiac glycosides. Scillaren-A and Scillaren-B are the major constituents. The drug also contains glucoscillaren A and enzyme Scillarenase . The other contents of drug are mucilage (about 40 %) and calcium oxalate

58. Chemical tests for Indian Squill Mucilage + Corallin soda Reddish-purple stains Red stains Mesophyll + Iodine Water

59. Uses of Indian Squill ADULTERANTS Commercial samples of Indian squill in the market, are mixed with Scilla hyacintoiana . It is a potent cardiotonic without having any cumulative effect (unlike Digitalis ). It is mostly employed in small doses as an effective expectorant specially in chronic bronchitis and asthma. It shows some anti-cancer activity against human epidermoid carcinoma. It also has some diuretic properties

60. MARKETED PREPARATIONS OF CARDIAC GLYCOSIDES

61. • Generic name : Digoxin. • Brand name : Lanoxin, Lanoxicaps , Cardoxin , Digitek . • It is a cardiotonic glycoside obtained mainly from Digitalis lanata . • Digoxin has positive inotropic and negative chronotropic activity. • It is used to control ventricular rate in atrial fibrillation and in the management of congestive heart failure with atrial fibrillation. Marketed Preparations of Cardiac Glycosides Lanoxin Pediatric injection C ardoxin 0.25 mg tablets DIGOXIN

62. • Indications : Heart failure, atrial fibrillation • Dosage : In Adults- oral solution 0.05mg/mL , Injectable solution- 0.1mg/ml. • Contraindications : Digitalis toxicity, ventricular tachycardia/fibrillation, obstructive cardiomyopathy.  • Side effect : Extra beats, anorexia, nausea and vomiting. Diarrhoea in elderly, confusion, dizziness, drowsiness, restlessness, nervousness, agitation and amnesia, Marketed Preparations of Cardiac Glycosides Lanoxin 250 micrograms tablets

63. • Indications : Congestive heart failure , atrial flutter, atrial tachycardia, atrial fibrillation. • Dosage : Adult and child age 12 years and above, orally 1.2 to1.6 mg per day in divided dose. Child 1 to 2 year age 0.04 mg per kg weight. • Contraindications : Hypersensitivity, hypokalemia , pregnancy, lactation, hepatic disease, A.V. block, ventricular tachycardia. • Side effect : Hypersensitivity, hypokalaemia, pregnancy, lactation, hepatic disease, A.V. block, ventricular tachycardia. Marketed Preparations of Cardiac Glycosides Digoxin- Sandoz 0.25mg tablets DIGITOXIN DIGITOXIN is similar in structure and effects to digoxin. It is excreted via liver. It could be used in patients with poor or erratic kidney function. It is now rarely used.

64. • Action : Digoxin immune FAB do not allow digoxin to bind to the sites of action and reverse digoxin toxicity. • Indications : Digoxin or digitoxin toxicity. • Contraindications : Hypersensitivity, mild digoxin toxicity, children , pregnancy, lactation. • Side effects : Facial swelling, redness, rapid respiration rate, low cardiac output. Marketed Preparations of Cardiac Glycosides D igiFab DIGIBIND Digoxin Immune Fab Overdose or accumulation of digoxin causes digitalis toxicity. Signs and symptoms : anorexia, diarrhoea, nausea and vomiting, bradycardia, premature ventricular contractions, malaise, blurred vision, visual illusions

65. Dixin Tablets IP 0.25 mg Marketed Preparations of Cardiac Glycosides Digoxin Tablets, USP 0.125 mg

66. Digoxin Elixir .05mg/ml 60mL Marketed Preparations of Cardiac Glycosides Lanoxin Paediatric 50 mcg /ml Elixir Syrup

67. THANKS! Presented By: References: 1. Pharmacognosy by C.K. Kokate , A.P. Purohit, S.B. Gokhale 2. Pharmacy180.com 3.https://www.slideshare.net/priyankagoswami/glycosidesem-8 4. Essentials of medical pharmacology by K.D Tripathi Utkarsha Ambavane Suyash Ardalkar Aditi Bangar Ganesh Bhandare Moulika Chakrabortty Deepak Kahar Pratiksha Karde Pranali Katale Shweta Kothawale Alisha Maniyar Pragati Mishra Soham Patil Anil Yadav