ANTI-DIARRHOEAL HERBAL COMPOSITIONS

Abstract

The present synergistic herbal compositions comprising herbal ingredients extracted from at least two herbal species selected from the group consisting of Ocimum basilicum, Agathosma betulina, Cinnamomum cassia, Cinnamomum verum and Apium graveolens helps maintain the electrolyte and fluid balance by exerting its inhibitory secretory activity and by increasing the absorptive capacity of the intestine to prevent dehydration during an episode of infectious and non-infectious diarrhoea and in dehydration associated with various clinical and non-clinical conditions

Claims

1. A synergistic herbal composition having anti-diarrhoeal properties, comprising herbal ingredients extracted from at least two herbal species selected from the group consisting of Ocimum basilicum (basil), Agathosma betulina, Cinnamomum cassia (cassia), Cinnamomum verum (bark of Cinnamomum) and Apium graveolens (celery).

2. The synergistic herbal composition as claimed in claim 1, wherein the said herbal ingredient is an oleoresin or is an oil extracted from the said herbal species.

3. The synergistic herbal composition as claimed in claim 1, wherein the said oleoresin and essential oils is extracted from aerial or non-aerial parts selected from the group comprising the leaf, stem, roots, bark and seed of the said herbal species.

4. The synergistic herbal composition as claimed in claim 1, wherein the said herbal ingredients is present in a concentration ranging from 50 ml/Kg to 300 ml/Kg.

5. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; and (b) cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg.

6. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; and (b) cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg.

7. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; and (b) buchu leaf oil in a concentration ranging from 0.33 L/kg to 25 L/kg.

8. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (c) basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; and (d) celery oil in a concentration ranging from 0.0008 L/kg to 8 L/kg.

9. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (c) basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; and (d) basil oleoresin in a concentration ranging from 0.002 L/kg to 23 L/kg.

10. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (c) cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; and (d) cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg.

11. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (c) cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; and (d) buchu leaf oil in a concentration ranging from 0.33 L/kg to 25 L/kg.

12. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; and (b) celery seed oil in a concentration ranging from 0.0008 L/kg to 8 L/kg.

13. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; and (b) basil oleoresin in a concentration ranging from 0.002 L/kg to 23 L/kg.

14. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg; and (b) buchu leaf oil in a concentration ranging from 0.33 L/kg to 25 L/kg.

15. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg; and (b) celery seed oil in a concentration ranging from 0.0008 L/kg to 8 L/kg.

16. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg; and (b) basil oleoresin in a concentration ranging from 0.002 L/kg to 23 L/kg.

17. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) buchu leaf oil in a concentration ranging from 0.33 L/kg to 25 L/kg; and (b) celery seed oil in a concentration ranging from 0.0008 L/kg to 8 L/kg.

18. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) buchu leaf oil in a concentration ranging from 0.33 L/kg to 25 L/kg; and (b) basil oleoresin in a concentration ranging from 0.002 L/kg to 23 L/kg.

19. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) celery seed oil in a concentration ranging from 0.0008 L/kg to 8 L/kg; and (b) basil oleoresin in a concentration ranging from 0.002 L/kg to 23 L/kg.

20. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) Basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; (b) Cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; and (c) Cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg.

21. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) Basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; (b) Cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; (c) Cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg; and (d) Buchu leaf oil in a concentration ranging from 0.33 L/kg to 25 L/kg.

22. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) Basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; (b) Cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; (c) Cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg; (d) Buchu leaf oil. in a concentration ranging from 0.33 L/kg to 25 L/kg; and (e) Celery seed oil in a concentration ranging from 0.0008 L/kg to 8 L/kg.

23. The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; (a) Basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; (b) Cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; (c) Cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg; (d) Buchu leaf oil. in a concentration ranging from 0.33 L/kg to 25 L/kg; (e) Celery seed oil in a concentration ranging from 0.0008 L/kg to 8 L/kg; and (f) Basil oleoresin in a concentration ranging from 0.002 L/kg to 23 L/kg.

24. The synergistic herbal composition as claimed in any one of the preceding claims, wherein the said composition further optionally comprises electrolyte(s) selected from the group comprising sodium (Na.sup.+), Cl.sup. (Chloride), HCO.sub.3.sup., K.sup.+ (Potassium), H.sub.2PO.sub.4, HPO.sub.4.sup.2, Mg.sup.2+ (Magnesium), Ca.sup.2+ (calcium) and SO.sub.4.sup.2 (sulphate).

25. The synergistic herbal composition as claimed in claim 24, wherein the said electrolyte(s) is present in a concentration ranging from about 8 meq to about 160 meq of Na.sup.+, about 5 meq to about 160 meq of Cl.sup., about 1 meq to about 45 meq of HCO.sub.3.sup., about 0.5 meq to about 25 meq of K.sup.+, about 0.1 to about 20 meq of H.sub.2PO.sub.4.sup., about 0.1 meq to about 20 meq of HPO.sub.4.sup.2, about 0.1 meq to about 6 meq of Mg.sup.2+, about 0.1 to 6 meq Ca.sup.+2, and about 0.006 to about 6 meq of SO.sub.4.sup.2.

26. The synergistic herbal composition as claimed in claim 1, wherein the said composition having osmolarity ranging from 50 mOSm to 325 mOSm.

27. The synergistic herbal composition as claimed in claim 1, wherein the said composition is formulated as an oral rehydration solution (ORS), as a powder, syrup, suspension, tablets and intravenous fluid.

28. Use of the herbal composition comprising herbal ingredients extracted from at least one herbal species selected from the group consisting of Ocimum basilicum, Agathosma betulina, Cinnamomum cassia, Cinnamomum verum and Apium graveolens for the treatment of diarrhoea, diarrhoea related disorders and electrolyte and fluid imbalance by exhibiting the following mechanisms; (i) increasing the non-nutrient and nutrient dependent electrolyte absorption from matured and differentiated villus epithelial cells of the intestine by decreasing active chloride secretion from the crypt of the intestine; (ii) decreasing the concentration of intracellular secretagogues such as cAMP, cGMP, and Ca.sup.2+; (iii) directly inhibiting anion channels, or decreasing the expression of secretory anion channels from the membrane or by increasing the turn-over rate of channels on the brush border membrane; (iv) increasing electrolyte absorption by increasing the cell turn-over rate; and (v) increasing electrolyte absorption by increasing the height of villus.

29. Use of the herbal composition comprising herbal ingredients extracted from at least one herbal species selected from the group consisting of Ocimum basilicum, Agathosma betulina, Cinnamomum cassia, Cinnamomum verum and Apium graveolens as a rehydration formulation in infectious and non-infectious diarrhea, Crohn's disease, ulcerative colitis, radiation and chemotherapeutic drug induced gastrointestinal diseases, environmental enteropathy and malnutrition, renal diseases, exercise induced dehydration, dehydrationin old subjects, pre-eclampsia and eclampsia during pregnancy, nausea and vomiting during pregnancy bowel preparation for colonoscopy, nausea and vomiting in pregnancy, astronauts during extended space stay etc.

30. Use of the herbal composition as claimed in claims 28 and 29, wherein the the said composition is prepared in electrolyte(s) selected from the group comprising sodium (Na.sup.+), Cl.sup. (Chloride), HCO.sub.3.sup., K.sup.+ (Potassium), H.sub.2PO.sub.4, HPO.sub.4.sup.2, Mg.sup.2+ (Magnesium) Ca.sup.2+ (Calcium) and SO.sub.4.sup.2 (sulphate).

31. Use of the herbal composition as claimed in claims 28 and 29, wherein the concentration of the said herbal composition is ranging from 50 ml/Kg to 300 ml/Kg.

32. A method of treating a subject suffering from diarrhoea, diarrhoea related disorders and electrolyte and fluid imbalance by administering a therapeutically effective concentration of an herbal composition, the said method comprising herbal ingredients extracted from at least one herbal species selected from the group consisting of Ocimum basilicum, Agathosma betulina, Cinnamomum cassia, Cinnamomum verum and Apium graveolens.

33. A method of treating a subject by inducing rehydration in subjects suffering from exercise induced dehydration, dehydration, Crohn's disease, ulcerative colitis, radiation and chemotherapeutic drug induced gastrointestinal diseases by administering a therapeutically effective concentration of an herbal composition, the said method comprising herbal ingredients extracted from at least one herbal species selected from the group consisting of Ocimum basilicum, Agathosma betulina, Cinnamomum cassia, Cinnamomum verum and Apium graveolens.

34. The method of treating as claimed in claims 32 and 33, wherein the said composition is prepared in an electrolyte(s) solution selected from the group comprising sodium (Na.sup.+), Cl.sup. (Chloride), HCO.sub.3.sup., K.sup.+ (Potassium), H.sub.2PO.sub.4, HPO.sub.4.sup.2, Mg.sup.2+ (Magnesium), Ca.sup.2+ (Calcium) and SO.sub.4.sup.2 (sulphate).

35. The method of treating a subject as claimed in claims 33 and 34, wherein the said herbal composition is administered in a concentration of the herbal composition is ranging from 50 ml/Kg to 300 ml/Kg.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0035] FIG. 1 depicts the effect of (A) Buchu leaf oil, (B) Basil oil, (C) Cassia oil, (D) Cinnamon bark oil, (E) Celery seed oil and (F) Basil oleoresin on inherent anion secretion. The aforesaid herbal extracts decreased anion secretion (delta I.sub.sc) at 30 minutes, 45 minutes and 1 hour;

[0036] FIG. 2 depicts the effect of (A) Buchu leaf oil and (B) Basil oil on forskolin (cAMP) induced anion secretion. The aforesaid herbal extracts significantly decreased forskolin induced chloride secretion (delta I.sub.sc);

[0037] FIG. 3 depicts the effect of (A) Buchu leaf oil, (B) combination of buchu leaf oil and cassia oil, (C) cinnamon bark oil and celery seed oil and a (D) combination of buchu leaf oil, basil oleoresin, cassia oil, cinnamon bark oil and celery seed oil on forskolin induced anion secretion;

[0038] FIG. 4(a) depicts Western blot analysis showing the effect of basil oil on protein levels of CFTR (a cAMP-activated chloride channel) and SGLT1 (sodium-coupled glucose transporter 1) in the brush border membrane of small intestinal tissues from mice, both in the presence and absence of forskolin;

[0039] FIG. 4(b) depicts Western blot analysis showing the effect of cinnamon bark oil (CBO) on the protein levels of CFTR and SGLT1 in the brush border membrane isolated from mouse small intestine, both in the presence and absence of forskolin;

[0040] FIG. 4(c) depicts Western blot analysis showing the effect of the formulation on the protein levels of CFTR and SGLT1 during secretagogue induced diarrheal conditions (formulation comprises a combination of Basil oil, Cinnamon bark oil, Cassia oil and Buchu leaf oil);

[0041] FIG. 5(a) depicts the effect of basil oil (BO) on fluid absorption using in vitro loop assay; FIG. 5(b) depicts the effect of cinnamon bark oil (CBO) on net fluid absorption using in vitro loop assay to study its effect on basal and forskolin-stimulated conditions; FIG. 5(c) depicts the Effect of cassia oil (CO) on net fluid absorption in mouse small intestine: in vitro loop assay; FIG. 5(d) depicts the effect of buchu leaf oil (BLO) on fluid absorption in mouse small intestine using in vitro loop assay.

[0042] FIG. 6 depicts a representative saturation kinetic curve showing the method to arrive at the concentration of buchu leaf oil to inhibit specific channels responsible for electrolyte loss and therefore effective in increasing net fluid absorption

ABBREVIATIONS USED

[0043] Isc: short circuit current

[0044] Km: The Michaelis constant

[0045] Vmax: Maximum velocity

DETAILED DESCRIPTION OF THE INVENTION

[0046] Geographical Source of Biological Material Used in the Present Invention:

[0047] Basil oleoresin extracted from Ocimum basilicum is procured from India,

[0048] Buchu leaf oil extracted from Agathosma betulina is procured from South Africa,

[0049] Basil oil extracted from Ocimum basilicum is procured from India,

[0050] Cassia oil extracted from Cinnamomum cassia is procured from Indonesia,

[0051] Cinnamon bark oil extracted from Cinnamomum verum is procured from Sri Lanka, and Celery seed oil extracted from Apium graveolens is procured from Egypt.

[0052] The invention will now be described in detail in connection with certain preferred and optional embodiments, so that various aspects thereof may be more fully understood and appreciated.

[0053] In a preferred embodiment, the present invention provides an herbal composition comprising at least two herbal ingredients selected from the group consisting of Ocimum basilicum, Agathosma betulina, Cinnamomum cassia, Cinnamomum verum and Apium graveolens and a pharmaceutically acceptable carrier.

[0054] Accordingly, the herbal composition comprises at least one component selected from an oleoresin extracted from Ocimum basilicum, and essential oils selected from the group comprising buchu leaf oil is extracted from Agathosma betulina, Basil oil is extracted from Ocimum basilicum; Cinnamomum cassia oil, cassia oil from the bark of Cinnamomum verum and celery oil from the seed of Apium graveolens.

[0055] The present synergistic herbal composition comprises herbal ingredients in a concentration ranging from 50 ml/Kg to 300 ml/Kg.

[0056] The following are the embodiments that the present invention provides with regards to the herbal compositions employed herein.

[0057] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0058] (a) basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; and [0059] (b) cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg.

[0060] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0061] (a) basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; and [0062] (b) cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg.

[0063] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0064] (a) basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; and [0065] (b) buchu leaf oil in a concentration ranging from 0.33 L/kg to 25 L/kg.

[0066] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0067] (a) basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; and [0068] (b) celery oil in a concentration ranging from 0.0008 L/kg to 8 L/kg.

[0069] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0070] (a) basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; and [0071] (b) basil oleoresin in a concentration ranging from 0.002 L/kg to 23 L/kg.

[0072] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0073] (a) cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; and [0074] (b) cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg.

[0075] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0076] (a) cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; and [0077] (b) buchu leaf oil in a concentration ranging from 0.33 L/kg to 25 L/kg

[0078] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0079] (a) cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; and [0080] (b) celery seed oil in a concentration ranging from 0.0008 L/kg to 8 L/kg.

[0081] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0082] (a) cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; and [0083] (b) basil oleoresin in a concentration ranging from 0.002 L/kg to 23 L/kg.

[0084] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0085] (a) cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg; and [0086] (b) buchu leaf oil in a concentration ranging from 0.33 L/kg to 25 L/kg.

[0087] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0088] (a) cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg; and [0089] (b) celery seed oil in a concentration ranging from 0.0008 L/kg to 8 L/kg.

[0090] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0091] (a) cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg; and [0092] (b) basil oleoresin in a concentration ranging from 0.002 L/kg to 23 L/kg.

[0093] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0094] (a) buchu leaf oil in a concentration ranging from 0.33 L/kg to 25 L/kg; and [0095] (b) celery seed oil in a concentration ranging from 0.0008 L/kg to 8 L/kg.

[0096] The synergistic herbal composition as claimed in claim 1, wherein the said composition comprises; [0097] (a) buchu leaf oil in a concentration ranging from 0.33 L/kg to 25 L/kg; and [0098] (b) basil oleoresin in a concentration ranging from 0.002 L/kg to 23 L/kg.

[0099] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0100] (a) celery seed oil in a concentration ranging from 0.0008 L/kg to 8 L/kg; and [0101] (a) basil oleoresin in a concentration ranging from 0.002 L/kg to 23 L/kg.

[0102] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0103] (a) Basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; [0104] (b) Cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; and [0105] (c) Cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg.

[0106] The aforesaid composition comprising three herbal ingredients extracted from Ocimum basilicum, Agathosma betulina, Cinnamomum cassia, Cinnamomum verum and Apium graveolens is formulated in varying combinations in appropriated concentrations.

[0107] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0108] (a) Basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; [0109] (b) Cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; [0110] (c) Cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg; and [0111] (d) Buchu leaf oil in a concentration ranging from 0.33 L/kg to 25 L/kg.

[0112] The aforesaid composition comprising the four herbal ingredients extracted from Ocimum basilicum, Agathosma betulina, Cinnamomum cassia, Cinnamomum verum and Apium graveolens is formulated in varying combinations in appropriated concentrations.

[0113] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0114] (a) Basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; [0115] (b) Cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; [0116] (c) Cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg; [0117] (d) Buchu leaf oil. in a concentration ranging from 0.33 L/kg to 25 L/kg; and [0118] (e) Celery seed oil in a concentration ranging from 0.0008 L/kg to 8 L/kg.

[0119] The aforesaid composition comprising the five herbal ingredients extracted from Ocimum basilicum, Agathosma betulina, Cinnamomum cassia, Cinnamomum verum and Apium graveolens is formulated in varying combinations in appropriated concentrations.

[0120] In an embodiment, the present invention provides a synergistic herbal composition, wherein the said composition comprises; [0121] (a) Basil oil in a concentration ranging from 0.027 L/kg to 57 L/kg; [0122] (b) Cinnamon bark oil in a concentration ranging from 0.33 L/kg to 22 L/kg; [0123] (c) Cassia oil in a concentration ranging from 0.67 L/kg to 50 L/kg; [0124] (d) Buchu leaf oil. in a concentration ranging from 0.33 L/kg to 25 L/kg; [0125] (e) Celery seed oil in a concentration ranging from 0.0008 L/kg to 8 L/kg; and [0126] (f) Basil oleoresin in a concentration ranging from 0.002 L/kg to 23 L/kg.

[0127] The individual anion secretion inhibitory effect of the components of the herbal composition, including Buchu leaf oil, basil oil, cassia oil, cinnamon bark oil, celery seed oil and basil oleoresin is demonstrated in FIG. 1. Each of the active agents cause decreased anion secretion within a minimum time duration of 30 minutes.

[0128] In another preferred embodiment, the present invention provides an herbal composition further comprising electrolytes selected from the group comprising sodium (Na.sup.+), Cl.sup. (Chloride), HCO.sub.3.sup. (bicarbonate), K.sup.+ (Potassium), H.sub.2PO.sub.4.sup., HPO.sub.4.sup.2 (phosphate ions), Mg.sup.2+ (Magnesium), Ca.sup.2+ (calcium) and SO.sub.4.sup.2 (sulphate).

[0129] Accordingly, the herbal composition comprises one or more of the electrolytes in a concentration ranging from about 8 meq to about 160 meq of Na.sup.+, about 6 meq to about 156 meq of Cl.sup., about 1 meq to about 45 meq of HCO.sub.3.sup., about 0.5 meq to about 25 meq of K.sup.+, about 0.1 to about 20 meq of H.sub.2PO.sub.4.sup., about 0.1 meq to about 20 meq of HPO.sub.4.sup.2, about 0.1 meq to about 6 meq of Mg.sup.2+, about 0.1 to 6 meq Ca.sup.+2, about 0.006 to about 6 meq of SO.sub.4.sup.2.

[0130] More preferably, the present herbal composition comprises 75 meq of Na.sup.+, 70 meq of Cl.sup., 25 meq of HCO.sub.3.sup., 20 meq of K.sup.+, 0.4 meq of H.sub.2PO.sub.4.sup., 2.4 meq of HPO.sub.4.sup.2, 1.2 meq of Mg, 1.2 meq Ca and 1.3 meq of SO.sub.4.sup.2.

[0131] In an embodiment, the present invention provides the herbal composition to have pH ranging from 3 to 7.6.

[0132] In a further embodiment, the present invention provides the herbal composition having osmolarity ranging from about 50 mOSm to about 325 mOSm.

[0133] In another preferred embodiment, the present invention provides a synergistic herbal composition comprising active ingredients selected from the group consisting of oleoresin of Ocimum basilicum, oil from leaves of Agathosma betulina, oil extracted from Cinnamomum cassia, oil extracted from bark of Cinnamomum verum and oil extracted from seeds of Apium graveolens and a pharmaceutically acceptable carrier.

[0134] The anion secretion inhibitory activity of the synergistic composition comprising combination of buchu leaf oil, basil oleoresin cassia oil, cinnamon bark oil and celery seed oil against forskolin-stimulated anion secretion is demonstrated in FIG. 3. In an episode of diarrhoea, cAMP activates CFTR channel incorporation and opening of these channels on the brush border membrane of villus epithelial cells, subsequently causing anion secretion. Anion secretion causes electrolyte and fluid loss leading to diarrhoea. The combination of buchu leaf oil, basil oleoresin cassia oil, cinnamon bark oil and celery seed oil resulted in further inhibition of forskolin-stimulated anion secretion (151.943.0 A/cm.sup.2 vs 73.527.2 A/cm.sup.2) (FIG. 3(D)). Therefore, these results reflect the synergistic anion secretion inhibitory activity of the herbal composition of the present invention against secretagogues such as cAMP, and cGMP involved in the occurrence of diarrhoea.

[0135] In another embodiment, the present invention provides an herbal composition having anti-diarrhoeal properties, wherein the said composition is formulated as an oral rehydration solution (ORS), as a powder, syrup, paste, suspension, tablets and intravenous fluid.

[0136] Accordingly, the present herbal composition can be administered via the oral or intravenous route.

[0137] In yet another embodiment, the present invention provides the herbal composition to further comprise pharmaceutically acceptable additives selected from the group comprising of sweeteners, flavouring agents, masking agents, colorants, preservatives, excipients, gelling agents, oligosaccharides, vitamins, dietary supplements, natural fruit extracts, amino acids and combinations thereof.

[0138] In an optional embodiment, the present invention provides the herbal composition comprising glucose.

[0139] In another optional embodiment, the present invention provides the herbal composition comprising a zinc source, most preferably elemental zinc.

[0140] In yet another preferred embodiment, the present invention provides a method of treating diarrhoea, diarrhoea related disorders and electrolyte and fluid imbalance the said method comprising administering an herbal composition comprising at least one herbal ingredient selected from the group consisting of Basil oleoresin, Buchu leaf oil, Basil oil, Cassia oil, Cinnamon bark oil and Celery seed oil and at least one electrolyte to an individual exhibiting symptoms of diarrhoea.

[0141] Accordingly, the present synergistic herbal composition can be administered to correct electrolyte and fluid imbalance, infectious and non-infectious diarrhoea, acute or chronic dehydration, antibiotics-induced diarrhoea, drug-induced diarrhoea, environmental enteropathy, diarrhoea secondary to food allergies, stress-induced diarrhoea, acute shock, hypovolemia, acute severe diarrhoea, hypotension, muscle cramps, Inflammatory Bowel Diseases (IBD) (Crohn's disease, ulcerative colitis), renal diseases, nausea and vomiting.

[0142] The present invention provides use of herbal compositions comprising active ingredients selected from the group consisting of oleoresin of Ocimum basilicum, leaf oil of Agathosma betulina, oil extracted from Cinnamomum cassia, oil extracted from bark of Cinnamomum verum and oil extracted from seeds of Apium graveolens and a pharmaceutically acceptable carrier, for treating diarrhoea, diarrhoea related disorders and electrolyte and fluid imbalance.

[0143] The following examples, which include preferred embodiments, will serve to illustrate the practice of this invention, it being understood that the particulars shown are by way of example and for purpose of illustrative discussion of preferred embodiments of the invention.

EXAMPLES

Example 1: Plant Extracts Screened for Anti-Diarrhoeal Activity

[0144] The following active ingredients, plant extracts or compounds were screened initially using the electrophysiological approach and were ranked according to their anti-secretory activity, absorptive capacity or a combination of both, all in the absence of a secretagogue. The active agents/compounds tested include Basil oleoresin, Buchu leaf oil, Basil oil, Cassia oil, Cinnamon bark oil and Celery seed oil.

Example 2: Extraction Process of Essential Oils and Oleoresins

[0145] (a) Extraction Process for Essential Oils [0146] The plant material to be screened for anti-diarrhoeal activity including Ocimum basilicum, Agathosma betulina, Cinnamomum cassia, Cinnamomum verum and Apium graveolen were collected, pre-processed and were steam distilled separately. The water phase was separated and the oil was filtered. The oil was treated with anhydrous 5 mg/10 mL sodium sulphate and was again filtered. Quality control assays were performed and the final product was filtered to obtain the respective oil. The essential oils extracted from each of the plants were stored in sterile containers at room temperature.

[0147] (b) Extraction procedure for oleoresins [0148] Ocimum basilicum to be screened for anti-diarrhoeal activity was collected pre-processed and was subjected to solvent extraction. The extract is filtered and desolventised and was blended with additives if required. Quality control assay was performed. The final extract was filtered. The basil oleoresin extracted was stored in sterile containers at room temperature.

Example 3: Methodology to Assess the Functionality of the Active Ingredients for their Antidiarrheal Properties

[0149] Electrophysiological approach was used to identify if the agent under study have unstimulated or basal anti-secretory activity, unstimulated or basal absorptive capacity or a combination of both, all in the absence of secretagogues. Electrophysiological technique will help identifying active secretion and/or absorption in the presence/absence of secretagogues.

Example 4: Preparation of Formulations

[0150]

TABLE-US-00002 TABLE A Dose (l/kg) lower Higher dose Active ingredient dose studied studied ml/kg 1 Basil oil 1.6 3.4 2 Cinnamon bark oil 20 1.3 3 Cassia oil 40 3 4 Buchu leaf oil 20 1.5 5 Celery seed oil 0.05 0.5 6 Basil oleoresin 0.14 1.4

[0151] The formulation was first arrived by using 100% of the active agents at their respective K.sub.M and functional studies validating the effectiveness of the formulation. Thus, the active agents were used in a 1:1 ratio using 100% of respective K.sub.M. However, in separate experiments it was found that different ratios of the active agents in a given formulation could be used to decrease current and therefore increase fluid absorption. Thus, the active agents could be mixed to form a formulation using a range of their different ratios, e.g., buchu leaf oil and cassia oil could be mixed at a ratio of 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10. Similarly, the ratios of the two oils could be used in their reverse proportion such that, cassia oil and buchu leaf oil could be mixed at a ratio of 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10.

TABLE-US-00003 TABLE B Method for arriving at the concentration of each component used in the formulation: Active agents were used in a 1:1 ratio using 100% of respective KM. The ratio maintained when the agents were used in their lower and the higher concentration. The lowest concentration that resulted in a significant decrease in current was then used as the lower dose and correspondingly for each of the active agents. The highest concentration was similarly derived from the highest concentration of the substrate that resulted in the maximum reduction in current in the saturation kinetic analysis. E.g., basil oil gave a change in chloride secretion at a concentration of 0.008 L/5 mL. The lower dose for kg body weight was therefore calculated to be 1.6 L/kg body weight (0.008 * 200 mL). Cumulative Cumu- dose studied lative Concentration of each at lowest dose component/kg concentration studied Combination body weight (L/kg) at 1 + 2 1.6 L/kg, 20 L/kg 10.8 2.3 1 + 3 1.6 L/kg, 40 L/kg 20.8 3.2 1 + 4 1.6 L/kg, 20 L/kg 10.8 2.4 1 + 5 1.6 L/kg, 0.05 L/kg 0.82 2.0 1 + 6 1.6 L/kg, 0.14 L/kg 0.9 2.4 2 + 3 20 L/kg, 40 L/kg 30 2.1 2 + 4 20 L/kg, 20 L/kg 20 1.4 2 + 5 20 L/kg, 0.05 L/kg 10 0.9 2 + 6 20 L/kg, 0.14 L/kg 10 1.4 3 + 4 40 L/kg, 20 L/kg 30 2.3 3 + 5 40 L/kg, 0.05 L/kg 20 1.8 3 + 6 40 L/kg, 0.14 L/kg 20 2.2 4 + 5 20 L/kg, 0.05 L/kg 10 1 4 + 6 20 L/kg, 0.14 L/kg 10 1.5 5 + 6 0.05 L/kg, 0.14 L/kg 0.1 1 1+ 2 + 3 1.6 L/kg, 20 L/kg, 20.5 2.6 40 L/kg 1 + 2 + 3 + 4 1.6 L/kg, 20 L/kg, 20.4 2.3 40 L/kg, 20 L/kg 1 + 2 + 3 + 4 + 5 1.6 L/kg, 20 L/kg, 16.3 1.94 40 L/kg, 20 L/kg 0.05 L/kg 1 + 2 + 3 + 4 + 5 + 6 1.6 L/kg, 20 L/kg, 13.6 1.9 40 L/kg, 20 L/kg 0.05 L/kg, 0.14 L/kg The aforesaid values are obtained in studies carried out with mice. To obtain the dosage applicable for human administration, dose conversion from animal to human was performed by multiplying the values by 60 kg. indicates data missing or illegible when filed

[0152] The aforesaid formulations (individual oils or their combinations) were prepared and tested in an electrolyte solution containing electrolyte composition as shown below. [0153] 75 mM Sodium chloride [0154] 20 mM Potassium chloride [0155] 1.2 mM Calcium chloride [0156] 1.2 mM Magnesium chloride [0157] 25 mM Sodium bicarbonate [0158] 2.4 mM Potassium hydrogen phosphate [0159] 0.4 mM Potassium dihydrogen phosphate

[0160] It should be noted that oils can be effective when used alone or in combination in different electrolyte concentrations, a range of total osmolarity (50 mosm to 350 mosm) and range of pH (2.1 to 8.5).

Example 5: Dose Optimization by Saturation Curve Assay

[0161] Saturation kinetics assay was done to determine the km and Vmax of the active ingredients under study. The tissue is treated with increasing concentrations of the active ingredients and is incubated in the Ussing chamber for a particular interval of time. Short circuit current and conductance values were measured after each addition and a curve was plotted with concentration of compounds along X axis and percentage inhibition on the Y axis.

[0162] K.sub.m and V.sub.max is determined using the classical Michaelis-Menton equation. K.sub.m is the concentration of the substrate (isolate), which permits the transporter to achieve half V.sub.max (I.sub.sc). The substrate (isolate) exerts its effect on transporters responsible for absorption and/or secretion by activating or inhibiting the transporters. Activation or inhibition of a transporter or channel by a specific substrate (isolate) at a high K.sub.m, signifies a low affinity for the substrate, and requires a greater concentration of substrate to achieve V.sub.max. K.sub.m for Basil oleoresin, Buchu leaf oil, Basil oil, Cassia oil, Cinnamon bark oil and Celery seed oil determined by saturation curve analysis were 0.016, 0.006, 0.012, 0.008, 0.016 and 0.008 L/5 mL respectively. The concentrations of the individual oils used in the formulations was calculated from the km (1.75 l, 2.85 l, 2.21 l, 1.0 l and 0.60 l for Basil oleoresin, Buchu leaf oil, Basil oil, Cassia oil, Cinnamon bark oil and Celery seed oil respectively). The lowest concentration of the substrate that showed a decrease in short circuit current was used for arriving at the dose.

[0163] A representative saturation kinetic curve showing the method to arrive at the concentration of buchu leaf oil to inhibit specific channels responsible for electrolyte loss and therefore effective in increasing net fluid absorption is shown in FIG. 6. From the same saturation kinetic analysis, the lowest concentration of buchu leaf oil that was found to reduce the current (Isc) was used to arrive at the lowest concentration of the oil to inhibit the channel responsible for electrolyte loss. Similarly, highest concentration of the given substrate that resulted in maximum reduction in current was used to arrive at the higher dose for the given oil.

[0164] The dose range for basil oleoresin is 0.002 l/kg to 23 l/kg, buchu leaf oil is 0.33 l/kg to 25 l/kg, basil oil is 0.027 l/kg to 57 l/kg, cassia oil is 0.67 l/kg to 50 l/kg, cinnamon bark oil is 0.33 l/kg to 22 l/kg, celery seed oil is 0.0008 l/kg to 8 l/kg. Formulations were made using appropriate proportions of the essential oil according to their activity. Saturation kinetics of each of the individual oils that showed significant decrease in anion secretion were conducted to determine the k.sub.m and V.sub.max. K.sub.m was used to determine the optimum concentration of the individual oils in the formulation.

Example 6: In Vitro Loop Assay: Ileal Sac Assays

[0165] The lowest concentrations of the oils were used for in vitro loop assay. Tissues were exposed to the individual oils at a concentration ranging from 0.001 L to 5 L in Ussing chamber. The oils that showed a more potent activity in functional and molecular studies were selected for the loop assay. The lowest concentration that showed a measurable decrease in short circuit current was used for the loop assay (buchu leaf oil 0.1 L, cassia oil 0.2 L, basil oil 0.008 L, cinnamon bark oil 0.1 L). Combination of oils were used into a formulation using different ratios of the active agents. The formulation was first arrived by using 100% of the active agents at their respective K.sub.M and functional studies validating the effectiveness of the formulation. Thus, the active agents were used in a 1:1 ratio using 100% of respective K.sub.M. However, in separate experiments it was found that different ratios of the active agents in a given formulation could be used to decrease current and therefore increase fluid absorption. Thus, the active agents could be mixed to form a formulation using a range of their different ratios, e.g., buchu leaf oil and cassia oil could be mixed at a ratio of 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10. Similarly, the ratios of the two oils could be used in their reverse proportion such that, cassia oil and buchu leaf oil could be mixed at a ratio of 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10. in the same concentration was used for the formulation. Ex vivo studies were performed with ileal segments approximately 10 cm in length that were ligated at one end and then filled with 200 l of Ringer solution, Ringer solution containing forskolin (10 M) or Ringer solution containing forskolin plus an agent under study (e.g. buchu leaf oil). The ileal segment was then tied at the other end to form a sac and incubated in Ringer solution bubbled with 95% 02 and 5% CO.sub.2. The beaker was slowly agitated in a water bath at 37 C. After 60-minute incubation, the sacs were removed from the beaker. The volume of the fluid in the ileal sac was measured, and the tissues rapidly cooled. The percentage of net fluid absorption was calculated using the formula (I-O)/I, where I=volume of fluid added to the lumen and O=volume of fluid in the lumen at the end of incubation time (FIG. 5). Mucosa was scraped out from the intestine and brush border membrane was isolated from the tissues. The brush border membrane was used further for western blot analysis.

Example 7: Functional Assay of Combination of Compounds for their Anti-Secretory Activity in the Presence/Absence of Secretagogues

[0166] The effect of buchu leaf oil with cassia oil and the combination of buchu leaf oil, basil oleoresin cassia oil, cinnamon bark oil and celery seed oil on forskolin-stimulated anion (Cl.sup. and HCO.sub.3.sup.) secretion is given in FIG. 2. It is noted that buchu leaf oil resulted in a significant decrease in forskolin-stimulated anion secretion (25.78.3 A/cm.sup.2 vs 40.46.7 A/cm.sup.2), but the combination of buchu leaf oil and cassia oil resulted in a far greater inhibition of the forskolin-stimulated chloride secretion (132.012.6 A/cm.sup.2 vs 62.319.9 A/cm.sup.2). Also, combination of cinnamon bark oil with celery seed oil resulted in significant decrease in forskolin-stimulated anion secretion (104.115.4 A/cm.sup.2 vs 75.813.0 A/cm.sup.2). Similarly, the combination of buchu leaf oil, basil oleoresin cassia oil, cinnamon bark oil and celery seed oil resulted in further inhibition of forskolin-stimulated anion secretion (151.943.0 A/cm.sup.2 vs 73.527.2 A/cm.sup.2) (FIG. 3).

Example 8

[0167] Western blot analysis was done to determine the expression of transporter involved in nutrient dependent and independent electrolyte absorption. Expression of secretory channels CFTR was found to be reduced in presence of the extracts and formulation. Animals at interdigestive phase and digestive phase were used to study the effect of oils on the expression of the transporter SGLT1 during nutrient independent and nutrient dependent phase respectively. Some of the oils increased the expression of SGLT1 and some decreased SGLT1 depends on the phase, whether digestive or inter-digestive (Representative data shown; FIGS. 4(a), (b) and (c)).

Example 9: The Ussing Chamber System

[0168] The present inventors have employed a functional approach for the validation of plant ingredients/compounds for their anti-secretory activity in Ussing chamber. The Ussing chamber provides an in vitro physiological system to measure the transport of ions, nutrients, and drugs across various epithelial tissues. The Ussing chamer system consists of two functional unitsThe chamber itself and the electric circuitry. Each chamber was filled with the physiological buffer solution, which is continuously aerated and connected with a circulating water bath to maintain the physiological pH and temperature. The electrical circuitry allows the measurement of parameters like short-circuit current, trans-epithelial conductance, voltage, impedence and capacitance. The Ussing chamber system also contains a software support system for data collection (Acquire and Analyze software). Silver/silver chloride voltage and current electrodes are used for the measurement of electrical parameters.

[0169] Mice intestinal epithelial tissue was collected by sharp dissection and is opened longitudinally along the entire length. The tissue is cut into 8 segments so that the segments form the entire length of the intestine (from jejunum to illeum) are included in the experiments. The 8 tissues are placed in the 8 channels of the Ussing chamber in a sequential order (jejunum to illeum). The electric circuitry is voltage clamped to zero, to measure the active transport of ions across the membrane. Data is captured using Acquire and Analyze software version 2.3.

Example 10: Agents that Decrease Active Chloride Secretion

[0170] Oleoresins obtained from solvent extraction process and essential oils obtained from steam distillation process were first screened for anti-secretory activity. Oleoresins/oils that showed activity/effectiveness was then ranked based on their anti-secretory activity measured using the electrophysiological approach mentioned. The compounds with the highest unstimulated or basal anti-secretory activity were therefore taken up for further evaluation. Compounds which decreased active chloride secretion/Antidiarrheal agents are enlisted in the table herein below.

TABLE-US-00004 Components of the herbal composition 1 Oleoresin Basil oleoresin 2 Oils Buchu leaf oil 3 Basil oil 4 Cassia oil 5 Cinnamon bark oil 6 Celery seed oil

[0171] The plant extracts were studied for the anti-secretory activity in the presence of a secretagogue such as cAMP. In a diarrhoea episode, increase in intracellular cAMP activates CFTR channel incorporation and opening of these channels on the brush border membrane of villus epithelial cells and subsequently causing anion secretion. Anion secretion causes electrolyte and fluid loss leading to secretory diarrhoea. The extracts screened in the present invention were determined to possess excellent anti-secretory property in the presence of a secretagogues, cAMP induced anion secretion was produced by using forskolin. Representative data of agents that decreased forskolin induced chloride secretion is given in FIG. 2.

Example 10(A): Synergy of the Present Composition

[0172] (i) Fluid Absorption: [0173] The formulation comprising of basil oil, cinnamon bark oil, cassia oil, buchu leaf and celery seed oil at a concentration as described in Table 1, resulted a net fluid absorption of 21 L/cm/hr when compared to 5.8 L/cm/hr, 31.8 L/cm/hr, 11.2 L/cm/hr, 2.7 L/cm/hr and 7.6 L/cm/hr for basil oil, cinnamon bark oil, cassia oil and buchu leaf oil respectively in small intestinal tissues incubated with forskolin. [0174] The formulation comprising of basil oil, cinnamon bark oil, cassia oil, buchu leaf oil, celery seed oil and basil oleoresin at a concentration as described in Table 1, resulted in a net fluid absorption of 11.6 L/cm/hr when compared to 5.8 L/cm/hr, 31.8 L/cm/hr, 11.2 L/cm/hr, 2.7 L/cm/hr, 7.6 L/cm/hr and 8 L/cm/hr for basil oil, cinnamon bark oil, cassia oil, buchu leaf oil, celery seed oil and basil oleoresin respectively in small intestinal tissues incubated with forskolin.

[0175] (ii) Effect of the Present Formulation on Protein Levels of CFTR and SGLT1 [0176] For studying synergistic effects, half the dose for each of the compound that showed an effective reduction in forskolin-stimulated increase in short circuit current was used (Data not shown). In order to study the synergy between basil oil and cinnamon bark oil, the said oils were mixed at a concentration of 0.8 ((1.6 L/kg)/2) and 10 ((20 L/kg)/2) L respectively. [0177] When two or more compounds were used together in a formulation, the effect was considered to be synergistic when the desired function was equal or more than what was obtained when used alone or one or more of the compounds neutralized the negative effect it may have in nutrient absorption during digestive or inter-digestive phase (FIGS. 4(a) and 4(b) compared to FIG. 4(c)). Incubating the small intestine with basil oil decreased SGLT1 protein levels suggesting that basil oil decreased glucose stimulated sodium absorption necessitating the importance for including other extracts or agents that will increase SGLT1 protein levels on the brush border membranes when used together as a formulation. Thus the formulation includes extracts or agents that cumulatively increased SGLT1 protein levels on the membrane and their function by demonstrating increased glucose stimulated increase in short circuit current (indicating increased electrolyte and glucose absorption). The reason for including basil oil in the formulation was because, when used together with the formulation, basil oil was found to be most effective in decreasing forskolin stimulated chloride secretion (FIG. 3 D). It also decreased unstimulated and secretagogue induced chloride secretion when used alone (FIGS. 1 B and 2 B). Western blot analysis for SGLT1 showed that forskolin stimulated an increase in SGLT1 expression (FIG. 4 A).

Example 10(B)

[0178] Forskolin stimulated CFTR protein levels decreased in the brush border membrane in mouse intestinal tissues treated with formulation (A combination of 1, 2, 3 and 4) (FIG. 4(c)). These observations agreed very well with electrophysiological observations that the formulation decreased forskolin stimulated increase in current (FIG. 3D). Therefore this formulation can be effectively used for treating secretory diarrhea that is associated with significant electrolyte and fluid loss. In vitro loop assay to measure net fluid movement showed that the formulation increased net fluid absorption when compared to control or basal conditions, indicating decreased electrolyte loss under basal conditions and therefore better hydration. The formulation comprising of basil oil, cinnamon bark oil, cassia oil and buchu leaf oil at a concentration as described in Table 1, resulted in a 4 fold increase in net fluid absorption in small intestinal tissues incubated with forskolin. Also, FSK induced increase in CFTR expression decrease when treated with the formulation, indicating that the formulation can effectively decreased the secretagogue (cAMP) induced chloride secretion. Therefore the formulation could be used in disease conditions that are associated with increased secretagogue-induced electrolyte loss, such as in infectious diarrhea. Western blot analysis for SGLT1 showed that the formulation in the presence of secretagogue further enhanced SGLT1 protein levels on the brush border membrane. Therefore, the formulation can be used to enhance electrolyte and nutrient absorption during diarrheal diseases, associated with increased electrolyte loss.