WO2011080721A2

WO2011080721A2 - A herbal composition as hepatoprotective and treatment for liver disorders

Info

Publication number: WO2011080721A2
Application number: PCT/IB2011/000002
Authority: WO
Inventors: Rangesh Paramesh; Uddagiri Venkanna Babu; Ekta Saxena
Original assignee: Himalaya Global Holdings Ltd.
Priority date: 2010-01-01
Filing date: 2011-01-01
Publication date: 2011-07-07
Also published as: WO2011080721A3

Abstract

The invention relates to a herbal composition, more particularly, a composition comprising extracts of Eclipta prostrata whole parts and/or Cichorium intybus seeds and/or Andrographis paniculata whole parts and/or Fumaria indico whole parts and/or Oroxylum indicum leaves and/or Ailanthus excelsa leaves for hepatoprotective and methods of treating liver disorders such as jaundice, cirrhosis, non-alcoholic fatty liver disease etc. and other liver disorders due to infection and chemotherapy in humans using said natural herbal composition.

Description

A HERBAL COMPOSITION AS HEPATOPROTECTIVE AND TREATMENT

FOR LIVER DISORDERS

FIELD OF THE INVENTION

This invention relates to a herbal composition having hepatoprotective activity. More particularly, the invention relates to a composition comprising extracts of Eclipta prostrata whole parts and/or Cichorium intybus seeds and/or Andrographis paniculata whole parts and/or Fumaria indica whole parts and/or Oroxylum indicum leaves and/or Ailanthus excelsa leaves and a pharmaceutically acceptable carrier, methods of obtaining the same, pharmaceutical formulations thereof and methods of using the composition as hepatoprotective which is useful in treating liver disorders such as jaundice, cirrhosis, non-alcoholic fatty liver disease etc. and other liver disorders due to infection and chemotherapy in humans using said natural herbal composition.

BACKGROUND OF THE INVENTION

Day by day we are identifying new and more powerful agents to prevent, treat, or retard infections and cure the diseases they cause. Billions of dollars are being spent every year in just identifying, locating, and developing these new drugs. However, till now effective therapies for many of the common disease like liver, heart, kidney etc. are still elusive.

At present, most patients are treated with drugs that are very effective over a short term. However, they have very adverse effect on long term use. They are also very expensive and demonstrate many side effects and uncertain activities.

In comparison to the above, the herbal compositions are very effective in curing such diseases in a very efficient manner without any side effects. The herbal composition prepared for liver helps in restoring the metabolic efficiency of the liver by protecting the hepatic parenchyma and promoting hepatocellular regeneration. It also helps in preventing the loss of functional integrity of the cell membrane and in restoring the hepatic function in infective hepatitis.

SUMMARY OF THE INVENTION This invention relates to herbal composition which is useful in treating liver disorders such as jaundice, cirrhosis, non-alcoholic fatty liver disease etc. and other liver disorders. More particularly, the invention relates to a herbal composition comprising extracts of Eclipta prostrate whole parts and/or Cichorium intybus seeds and/or Andrographis paniculata whole parts and/or Fumaria indica whole parts and/or Oroxylum indicum leaves and/or Ailanthus excelsa leaves and a pharmaceutically acceptable carrier, methods of obtaining the same, pharmaceutical formulations thereof and methods of using the composition as hepatoprotective agent.

Further, the present invention relates to the selection and identification of the herbs and obtaining the extract by subjecting the same to different extraction techniques including conventional solvent extraction and super critical fluid extraction. The bioassay guided fractionation of the extract or combination thereof to identify the active markers or active fraction and to develop effective and safe composition for the use in human beings and animals as hepatoprotective agent and treatment for all kinds of liver disorders such as jaundice, cirrhosis, non-alcoholic fatty liver disease and other liver disorders due to infection and chemotherapy in humans.

BRIEF DESCRIPTION OF DRAWINGS

FIG.l depicts the effect of N P-042/ERNP variants on SGPT level of rats in an experimental model of carbon tetrachloride-induced hepatic damage in rats.

FIG.2 depicts the effect of N P-042/ERN P variants on SGOT level of

experimental model of carbon tetrachloride-induced hepatic damage in rats. FIG.3 depicts the effect of N P-042/ERNP variants on liver glycogen content of rats in an experimental model of carbon tetrachloride-induced hepatic damage in rats. FIG. depicts the comparative evaluation of NP-42 variants for Hepatoprotective activity in an experimental model of thioacetamide induced hepatic damage in rats.

FIG.5 depicts the comparative evaluation of NP-42 variants for Hepatoprotective activity in an experimental model of thioacetamide induced hepatic damage in rats.

FIG.6 depicts the comparative evaluation of NP-42 variants for Hepatoprotective activity in an experimental model of thioacetamide induced hepatic damage in rats.

FIG.7 depicts the comparative evaluation of NP-42 variants in an experimental model of carbon tetrachloride -induced hepatic damage in rats.

FIG.8 depicts the comparative evaluation of N P-42 variants in an experimental model of carbon tetrachloride induced hepatic damage in rats. FIG.9 depicts the comparative evaluation of NP-42 variants in an experimental model of carbon tetrachloride induced hepatic damage in rats.

EMBODIMENT OF THE INVENTION In an embodiment of proposed invention a herbal composition having hepatoprotective activity comprises effective amounts of extracts of Eclipta prostrato, Cichorium intybus, Andrographis paniculata, Fumaria indica, Ailanthus excelsa and suitable pharmaceutically acceptable carriers and additives. In another embodiment of the invention the herbal extracts are obtained either by separate extraction or from the blend of Eclipta prostrata, Cichorium intybus, Andrographis paniculata, Fumaria indica, Ailanthus excelsa parts. In another embodiment of the invention the herbal extracts are obtained from the specific parts of the herbs preferably whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Andrographis paniculata, whole parts of Fumaria indica and leaves of Ailanthus excelsa. In still another embodiment of the invention the blend of herbs comprises whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Andrographis paniculata, whole parts of Fumaria indica and leaves of Ailanthus excelsa in the ratio of 20:18:30: 15 :17 respectively. In still another embodiment of the invention a herbal composition having hepato- protective activity comprises effective amounts of extracts of Eclipta prostrata, Cichorium intybus, Andrographis paniculata, Fumaria indica, Oxoxylum indicum and suitable pharmaceutically acceptable carrier and additives. In still another embodiment of the invention the herbal extracts are obtained either by separate extraction or from the blend of Eclipta prostrata, Cichorium intybus, Andrographis paniculata, Fumaria indica, Oxoxylum indicum parts.

In yet another embodiment of the invention the herbal extracts are obtained from the specific pa rts of the herbs preferably whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Andrographis paniculata, whole parts of Fumaria indica and leaves of Oxoxylum indicum. In still another embodiment of the invention the blend of herbs comprises whole parts of Eclipta prostrota, seeds of Cichorium intybus, whole parts of Andrographis paniculata, whole parts of Fumaria indica and leaves of Oxoxylum indicum in the ratio of 20:18:30:15:17 respectively. In still another embodiment of the invention the extracts are obtained by method selected from the group of percolation method, hot - soxlation method, super - critical - fluid method or enzyme extraction method.

In still another embodiment the said extracts are obtained by percolation method comprising the following steps of: . a. Shade drying the material; b. Pulverizing the material obtained in step a. to coarse powder; c. Placing the powdered material in different percolators and extracting with n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1:1), methanol and water (1:1) and water at room temperature for 24 h to 48 h.; and d. Filtering the plant extracts and concentrating to dryness using a rotatory evaporator or on steam bath at optimum temperature under reduced pressure.

In still another embodiment the said extracts are obtained by hot-soxlation method comprising the steps of:

a. Placing the coarse powdered material in each soxlator using solvents n- hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1:1), methanol and water ( 1 :1) and water at refluxing temperature of each solvent;

b. recycling the process until extraction is completed; and

c. filtering the plant extracts and concentrating to dryness using rotatory evaporator or on steam bath at optimum temperature.

In still another embodiment the said extracts are obtained by super - critical - fluid extraction method comprising the steps of: a. Shade drying the material; b. Pulverizing the dried material to coarse powder; c. Placing the material in a SCF extractor at the temperature ranging from 40-50 °C at high pressure range of 300-350 bar using carbon dioxide as super critical fluid for extraction upto 4 to 6 hours; and d. Collecting the extract in the collection vessel and evaporating at room temperature to remove any further residues of carbon dioxide.

In another embodiment the said extracts are obtained by enzyme extraction method comprising the steps of: a. Shade drying the material; b. Pulverizing the dried material to coarse powder; c. Placing the material in Stainless Steel container and adding 0.5% to 5% of cellulase and pectinase enzyme combination in 4 volumes of distilled water; d. Processing the enzyme extraction at the temperature of 55°C to 60°C under occasional stirring upto 4-6 hours; and e. Filtering the extract and concentrating the same to dryness on rotatory evaporator or on steam bath at optimum temperature under reduced pressure.

In another embodiment the herbal composition is in the form of a tablet, capsule, syrup, injection or any other suitable form.

DETAILED DESCRIPTION OF THE INVENTION

An important aspect of the present invention is to provide a herbal composition which is useful in treating liver disorders such as jaundice, cirrhosis, non-alcoholic fatty liver disease etc. and other liver disorders. More particularly, the invention relates to a herbal composition comprising extracts of Eclipto prostrata whole parts and/or Cichorium intybus seeds and/or Andrographis paniculata whole parts and/or Fumaria indica whole parts and/or Oroxylum indicum leaves and/or Ailanthus excelsa leaves and a pharmaceutically acceptable carrier, methods of obtaining the same, pharmaceutical formulations thereof and methods of using the composition as hepatoprotective.

Further, the present invention involves the selection and identification of the herbs and obtaining the extract by subjecting the same to different extraction techniques including conventional solvent extraction and super critical fluid extraction. The bioassay guided fractionation of the extract or combination thereof to identify the active markers or active fraction and to develop effective and safe composition for the use in human beings and animals as hepatoprotective agent and treatment for all kinds of liver disorders such as jaundice, cirrhosis, non-alcoholic fatty liver disease and other liver disorders due to infection and chemotherapy in humans. The following methodologies, examples, and studies of the composition follow the teachings of and illustrate the present invention :

Example 1

Preparation of extract from Eclipta prostrata by percolation method

The shade dried material of whole plant parts of Eclipta prostrata was pulverized to coarse powder and about 10 Kg each of powdered material placed in different percolators and extracted with n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1:1), methanol and water (1 :1) and water at room temperature for 24 h to 48 h., then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure.

Example 2

Preparation of extract from Eclipta prostrate by hot-soxlation method

The coarse powdered material of whole parts of Eclipta prostrata was subjected to hot- soxlation by placing 10 Kg of material in each soxlator using solvents n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1: 1), methanol and water (1:1) and water at refluxing temperature of each solvent and recycled the process until extraction is completed, then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperatu re.

All extracts such as n-hexane extract (EP-1), acetone extract (EP-2 ), ethyl alcohol extract (EP-3), methanol extract (EP-4), ethyl alcohol and water ( 1:1) extract (EP-5), methanol and water (1:1) extract (EP-6) and water extract (EP-7) prepared from the whole parts of Eclipta prostrata by percolation method or hot-soxlation method were subjected to HPTLC (High Performance Thin Layer Chromatography) and HPLC (High performance Liquid chromatography) in various mobile phases on precoated TLC plates (Merck) and ODS column for qualitative and quantitative estimation of marker compounds and active principles. Example 3

Preparation of extract from Eclipta prostrata by Enzyme extraction

The shade dried material of whole plant parts of Eclipta prostrata was pulverized to powder and about 10 Kg each of powdered material placed in Stainless Steel container and 0.5% to 5% of cellulase and pectinase enzyme combination was added in 4 volumes of distilled water. The enzyme extraction was processed at the temperature of 55°C to 60°C under occasional stirring upto 4-6 hours, and then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure.

Example 4

Preparation of extract from Eclipta prostrata by Super Critical Fluid Extraction

The shade dried material of whole plant parts of Eclipta prostrata was pulverized to coarse powder and about 100 Kg of powdered material placed in a SCF extractor at the temperature of 40-50C at high pressure of 300-350 bar using carbon dioxide as super critical fluid for extraction upto 4 to 6 hours and then the extract was collected in the collection vessel and evaporated at room temperature to remove any further residues of carbon dioxide. The extract thus obtained was free from any solvent residues and in highest pure form.

Example 5

Preparation of extract from Cichorium intybus by percolation method

The shade dried material of seeds of Cichorium intybus was pulverized to coarse powder and about 10 Kg each of powdered material placed in different percolators and extracted with n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water ( 1: 1), methanol and water ( 1:1) and water at room temperature for 24 h to 48 h., then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure.

Example 6

Preparation of extract from Cichorium intybus by hot-soxlation method

The coarse powdered material of seeds of Cichorium intybus was subjected to hot- soxlation by placing 10 Kg of material in each soxlator using solvents n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1 :1), methanol and water (1:1) and water at refluxing temperature of each solvent and recycled the process until extraction is completed, then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature.

All extracts such as n-hexane extract (Cl- l), acetone extract (CI-2 ), ethyl alcohol extract (CI-3), methanol extract (CI-4), ethyl alcohol and water ( 1: 1) extract (CI-5), methanol and water ( 1: 1) extract (CI-6) and water extract (CI-7) prepared from the whole parts of Cichorium intybus by percolation method or hot-soxlation method were subjected to HPTLC (High Performance Thin Layer Chromatography) and HPLC (High performance Liquid chromatography) in various mobile phases on precoated TLC plates (Merck) and ODS column for qualitative and quantitative estimation of marker compounds and active principles.

Example 7

Preparation of extract from Cichorium intybus by Enzyme extraction

The shade dried material of seeds of Cichorium intybus was pulverized to powder and about 10 Kg each of powdered material placed in Stainless Steel container and 0.5% to 5% of cellulase and pectinase enzyme combination was added in 4 volumes of distilled water. The enzyme extraction was processed at the temperature of 55°C to 60°C under occasional stirring upto 4-6 hours, and then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure.

Example 8

Preparation of extract from Cichorium intybus by Super Critical Fluid Extraction

The shade dried material of seeds of Cichorium intybus was pulverized to coarse powder and about 100 Kg of powdered material placed in a SCF extractor at the temperature of 40-50C at high pressure of 300-350 bar using carbon dioxide as super critical fluid for extraction upto 4 to 6 hours and then the extract was collected in the collection vessel and evaporated at room temperature to remove any further residues of carbon dioxide. The extract thus obtained was free from any solvent residues and in highest pure form.

Example 9

Preparation of extract from Andrographis paniculata by percolation method

The shade dried material of whole plant parts of Andrographis paniculata was pulverized to coarse powder and about 10 Kg each of powdered material placed in different, percolators and extracted with n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water ( 1: 1), methanol and water ( 1 : 1) and water at room temperature for 24 h to 48 h., then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure .

ExamplelO

Preparation of extract from Andrographis paniculata by hot-soxlation method

The coarse powdered material of whole parts of Andrographis paniculata was subjected to hot-soxlation by placing 10 Kg of material in each soxlator using solvents n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water ( 1: 1), methanol and water ( 1:1) and water at refluxing temperature of each solvent and recycled the process until extraction is completed, then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature.

All extracts such as n-hexane extract (AP-1), acetone extract (AP-2), ethyl alcohol extract (AP-3), methanol extract (AP-4), ethyl alcohol and water (1:1) extract (AP-5), methanol and water ( 1: 1) extract (AP-6) and water extract (AP-7) prepared from the whole parts of Andrographis panicuiata by percolation method or hot-soxlation method were subjected to HPTLC (High Performance Thin Layer Chromatography) and H PLC (High performance Liquid chromatography) in various mobile phases on precoated TLC plates (Merck) and ODS column for qualitative and quantitative estimation of marker compounds and active principles.

Example 11

Preparation of extract from Andrographis panicuiata by Enzyme extraction

The shade dried material of whole plant parts of Andrographis panicuiata was pulverized to powder and about 10 Kg each of powdered material placed in Stainless Steel container and 0.5% to 5% of cellulase and pectinase enzyme combination was added in 4 volumes of distilled water. The enzyme extraction was processed at the temperature of 55°C to 60°C under occasional stirring upto 4-6 hours, and then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure.

Examplel2

Preparation of extract from Andrographis panicuiata by Super Critical Fluid Extraction The shade dried material of whole plant parts of Andrographis panicuiata was pulverized to coarse powder and about 100 Kg of powdered material placed in a SCF extractor at the temperature of 40-50°C at high pressu re of 300-350 bar using carbon dioxide as super critical fluid for extraction upto 4 to 6 hours and then the extract was collected in the collection vessel and evaporated at room temperature to remove any further residues of carbon dioxide. The extract thus obtained was free from any solvent residues and in highest pure form. Example 13

Preparation of extract from Fumaria indica by percolation method

The shade dried material of whole plant parts of Fumaria indica was pulverized to coarse powder and about 10 Kg each of powdered material placed in different percolators and extracted with n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1:1), methanol and water (1:1) and water at room temperature for 24 h to 48 h., then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure.

Example 14

Preparation of extract from Fumaria indica by hot-soxlation method

The coarse powdered material of whole parts of Fumaria indica was subjected to hot- soxlation by placing 10 Kg of material in each soxlator using solvents n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1:1), methanol and water ( 1:1) and water at refluxing temperature of each solvent and recycled the process until extraction is completed, then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature.

All extracts such as n-hexane extract (Fl- l), acetone extract (FI-2), ethyl alcohol extract (FI-3), methanol extract (FI-4), ethyl alcohol and water ( 1: 1) extract (FI-5), methanol and water (1: 1) extract (FI-6) and water extract ( FI-7) prepared from the whole parts of Fumaria indica by percolation method or hot-soxlation method were subjected to HPTLC (High Performance Thin Layer Chromatography) and HPLC (High performance Liquid chromatography) in various mobile phases on precoated TLC plates (Merck) and ODS column for qualitative and quantitative estimation of marker compounds and active principles.

Example 15

Preparation of extract from Fumoria indica by Enzyme extraction

The shade dried material of whole plant parts of Fumario indica was pulverized to powder and about 10 Kg each of powdered material placed in Stainless Steel container and 0.5% to 5% of cellulase and pectinase enzyme combination was added in 4 volumes of distilled water. The enzyme extraction was processed at the temperature of 55°C to 60°C under occasional stirring upto 4-6 hours, and then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure.

Example 16

Preparation of extract from Fumoria indica by Super Critical Fluid Extraction

The shade dried material of whole plant parts of Fumaria indica was pulverized to coarse powder and about 100 Kg of powdered material placed in a SCF extractor at the temperature of 40-50°C at high pressure of 300-350 bar using carbon dioxide as super critical fluid for extraction upto 4 to 6 hours and then the extract was collected in the collection vessel and evaporated at room temperature to remove any further residues of carbon dioxide. The extract thus obtained was free from any solvent residues and in highest pure form.

Example 17

Preparation of extract from Oroxylum indicum by percolation method

The shade dried material of leaves of Oroxylum indicum was pulverized to coarse powder and about 10 Kg each of powdered material placed in different percolators and extracted with n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1: 1), methanol and water ( 1:1) and water at room temperature for 24 h to 48 h., then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure. Example 18

Preparation of extract from Oroxylum indicum by hot-soxlation method

The coarse powdered material of leaves of Oroxylum indicum was subjected to hot- soxlation by placing 10 Kg of material in each soxlator using solvents n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1:1), methanol and water (1 :1) and water at refluxing temperatu re of each solvent and recycled the process until extraction is completed, then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperatu re.

EP All extracts such as n-hexane extract (Ol-l), acetone extract (OI-2), ethyl alcohol extract (01-3), methanol extract (01-4), ethyl alcohol and water ( 1: 1) extract (01-5), methanol and water (1:1) extract (01-6) and water extract (01-7) prepared from the leaves of Oroxylum indicum by percolation method or hot-soxlation method were subjected to H PTLC (High Performance Thin Layer Chromatography) and HPLC (High performance Liquid chromatography) in various mobile phases on precoated TLC plates (Merck) and ODS column for qualitative and quantitative estimation of marker compounds and active principles.

Example 19

Preparation of extract from Oroxylum indicum by Enzyme extraction

The shade dried material of leaves of Oroxylum indicum was pulverized to powder and about 10 Kg each of powdered material placed in Stainless Steel container and 0.5% to 5% of cellulase and pectinase enzyme combination was added in 4 volumes of distilled water. The enzyme extraction was processed at the temperature of 55°C to 60°C under occasional stirring upto 4-6 hours, and then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure. Example 20

Preparation of extract from Oroxylum indicum by Super Critical Fluid Extraction

The shade dried material of leaves of Oroxylum indicum was pulverized to coarse powder and about 100 Kg of powdered material placed in a SCF extractor at the temperature of 40-50°C at high pressure of 300-350 bar using carbon dioxide as super critical fluid for extraction upto 4 to 6 hou rs and then the extract was collected in the collection vessel and evaporated at room temperature to remove any further residues of carbon dioxide. The extract thus obtained was free from any solvent residues and in highest pure form. Example 21

Preparation of extract from Herbal Blend by percolation method

The shade dried material of herbal blend of whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Andrographis poniculata, whole parts of Fumaria indica and leaves of Oxoxylum indicum in the ratio of 20:18:30:15: 17 respectively mixed and pulverized to coarse powder and about 10 Kg each of herbal blend placed in different percolators and extracted with n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1: 1), methanol and water ( 1:1) and water at room temperature for 24 h to 48 h., then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure .

Example 22

Preparation of extract from Herbal Blend by hot-soxlation method The coarse powdered material of herbal blend of whole parts of Eclipto prostrata, seeds of Cichorium intybus, whole parts of Andrographis paniculata, whole parts of Fumaria indica and leaves of Oxoxylum indicum in the ratio of 20: 18:30: 15:17 respectively was subjected to hot-soxlation by placing 10 Kg of herbal blend in each soxlator using solvents n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water ( 1:1), methanol and water ( 1:1) and water at refluxing temperature of each solvent and recycled the process until extraction is completed, then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature.

All extracts such as n-hexane extract (NP42-1), acetone extract (NP42-2 ), ethyl alcohol extract (NP42-3), methanol extract (N P42-4), ethyl alcohol and water ( 1 : 1) extract (NP42-5), methanol and water ( 1 : 1) extract (N P42-6) and water extract (N P42 -7) prepared from the herbal blend by percolation method or hot-soxlation method were subjected to H PTLC (High Performance Thin Layer Chromatography) and HPLC ( High performance Liquid chromatography) in various mobile phases on precoated TLC plates (Merck) and ODS column for qualitative and quantitative estimation of marker compounds and active principles. The water extract of herbal blend was prepared in the pilot scale batch from 100 Kg of herbal blend and the final extract was subjected to spray drying to obtain dry powder extract. This extract was coded as NP-042 and subjected to animal studies.

Example 23

Preparation of extract from herbal blend by Enzyme extraction

The shade dried material of herbal blend of whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Andrographis paniculata, whole parts of Fumaria indica and leaves of Oxoxylum indicum in the ratio of 20: 18:30: 15:17 respectively was pulverized to powder and about 10 Kg each of powdered herbal blend was placed in Stainless Steel container and 0.5% to 5% of cellulase and pectinase enzyme combination was added in 4 volumes of distilled ^"water. The enzyme extraction was processed at the temperature of 55°C to 60°C under occasional stirring upto 4-6 hours, and then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure. The yields of single and herbal blend enzyme extracts are mentioned in below table.

Table-1

Example 24

Preparation of extract from herbal blend by Super Critical Fluid Extraction

The shade dried material of herbal blend of whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Andrographis paniculata, whole parts of Fumaria indica and leaves of Oxoxylum indicum in the ratio of 20:18:30: 15: 17 respectively was pulverized to coarse powder and about 100 Kg of powdered herbal blend was placed in a SCF extractor at the temperature of 40-50°C at high pressure of 300-350 bar using carbon dioxide as super critical fluid for extraction upto 4 to 6 hours and then the extract was collected in the collection vessel and evaporated at room temperature to remove any further residues of carbon dioxide. The extract thus obtained was free from any solvent residues and in highest pure form.

Example 25

Preparation of extract from Ailanthus excelsa by percolation method;

The shade dried material of leaves of Ailanthus excelsa was pulverized to coarse powder and about 10 Kg each of powdered material placed in different percolators and extracted with n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water ( 1: 1), methanol and water (1: 1) and water at room temperature for 24 h to 48 h., then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure.

Example 26

Preparation of extract from Ailanthus excelsa by hot-soxlation method:

The coarse powdered material of leaves of Ailanthus excelsa was subjected to hot- soxlation by placing 10 Kg of material in each soxlator using solvents n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1 :1), methanol and water ( 1:1) and water at refluxing temperature of each solvent and recycled the process until extraction is completed, then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature.

All extracts such as n-hexane extract (AE-1), acetone extract (AE-2), ethyl alcohol extract (AE-3), methanol extract (AE-4), ethyl alcohol and water ( 1: 1) extract (AE-5), methanol and water (1:1) extract (AE-6) and water extract (AE-7) prepared from the leaves of Ailanthus excelsa by percolation method or hot-soxlation method were subjected to HPTLC (High Performance Thin Layer Chromatography) and HPLC (High performance Liquid chromatography) in various mobile phases on precoated TLC plates (Merck) and ODS column for qualitative and quantitative estimation of marker compounds and active principles.

Example 27

Preparation of extract from Ailanthus excelsa by Enzyme extraction

The shade dried material of leaves of Ailanthus excelsa was pulverized to powder and about 10 Kg each of powdered material placed in Stainless Steel container and 0.5% to 5% of cellulase and pectinase enzyme combination was added in 4 volumes of distilled water. The enzyme extraction was processed at the temperature of 55°C to 60°C under occasional stirring upto 4-6 hours, and then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure.

Example 28

Preparation of extract from Ailanthus excelsa by Super Critical Fluid Extraction

The shade dried material of leaves of Ailanthus excelsa was pulverized to coarse powder and about 100 Kg of powdered material placed in a SCF extractor at the temperature of 40-50°C at high pressure of 300-350 bar using carbon dioxide as super critical fluid for extraction upto 4 to 6 hours and then the extract was collected in the collection vessel and evaporated at room temperature to remove any further residues of carbon dioxide. The extract thus obtained was free from any solvent residues and in highest pure form.

Example 29

Preparation of extract from Herbal Blend by percolation method :

The shade dried material of herbal blend of whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Andrographis paniculata, whole parts of Fumaria indica and leaves of Ailanthus excelsa in the ratio of 20:18:30:15:17 respectively mixed and pulverized to coarse powder and about 10 Kg each of herbal blend placed in different percolators and extracted with n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1:1), methanol and water (1:1) and water at room temperature for 24 h to 48 h., then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature and under reduced pressure .

Example 30

Preparation of extract from Herbal Blend by hot-soxlation method:

The coarse powdered material of herbal blend of whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Andrographis paniculata, whole parts of Fumaria indica and leaves of Ailanthus excelsa in the ratio of 20:18:30:15:17 respectively was subjected to hot-soxlation by placing 10 Kg of herbal blend in each soxlator using solvents n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1 :1), methanol and water ( 1:1) and water at refluxing temperature of each solvent and recycled the process until extraction is completed, then plant extracts were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperature. All extracts such as n-hexane extract (NP42-8, also decoded as ERN P-167), acetone , extract (N P42-9, also decoded as ERNP-169 ), ethyl alcohol extract (N P42-10, also decoded as ERN P-168), methanol extract (NP42-11), ethyl alcohol and water ( 1: 1) extract (NP42-12, also decoded as NP42/A/123), methanol and water ( 1:1) extract (NP42-13) and water extract (NP42-14, also decoded as ERN P-172) prepared from the herbal blend by percolation method or hot-soxlation method were subjected to HPTLC (High Performance Thin Layer Chromatography) and HPLC (High performance Liquid chromatography) in various mobile phases on precoated TLC plates (Merck) and ODS column for qualitative and quantitative estimation of marker compounds and active principles.

The water extract of herbal blend (ERNP-172) was prepared in the pilot scale batch from 100 Kg of herbal blend and the final extract was subjected to spray drying to obtain dry powder extract. This extract was coded as NP-042/A and subjected to animal studies

Example 31

Preparation of extract from herbal blend by Enzyme extraction

The shade dried material of herbal blend of whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Andrographis paniculata, whole parts of Fumaria indica and leaves of Ailanthus excelsa in the ratio of 20:18:30:15: 17 respectively was pulverized to powder and about 10 Kg each of powdered herbal blend was placed in Stainless Steel container and 0.5% to 5% of cellulase and pectinase enzyme combination was added in 4 volumes of distilled water. The enzyme extraction was processed at the temperature of 55°C to 60°C under occasional stirring upto 4-6 hours, and then plant extracts (coded as ERNP-170) were filtered and concentrated to dryness on rotatory evaporator or on steam bath at optimum temperatu re and under reduced pressure. Example 32

Preparation of extract from herbal blend by Super Critical Fluid Extraction

The shade dried material of herbal blend of whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Andrographis paniculata, whole parts of Fumaria indica and leaves of Ailanthus excelsa in the ratio of 20:18:30: 15:17 respectively was pulverized to coarse powder and about 100 Kg of powdered herbal blend was placed in a SCF extractor at the temperature of 40-50C at high pressure of 300-350 bar using carbon dioxide as super critical fluid for extraction upto 4 to 6 hours and then the extract was collected in the collection vessel and evaporated at room temperature to remove any further residues of carbon dioxide. The extract thus obtained was free from any solvent residues and in highest pure form.

Example 33

Animal studies: Comparative evaluation of NP-042 variants in an experimental model of carbon tetrachloride and thioacetamide induced hepatic damage in rats

OBJECTIVE : The objective of the present study was to compare the hepatoprotective potential of N P-042/ERNP variants against carbontetrachloride and thioacetamide- induced hepatic damage in rats.

Studv-1

Evaluation of different NP-042/ERNP extracts in an experimental model of carbon tetrachloride-induced hepatic damage in rats:

Eighty male Wistar rats were divided in to eight groups ten each. The rats from groups I and II received DM water once a day orally at a dose of 10 ml/kg b.wt. and were served as normal and positive untreated controls respectively. Rats from groups III to VIII received N P-042/ERN P-167, NP-042/ERN P-168, N P-042/ERNP-169, NP-042/ERNP-170, NP-042/ERN P-171 and NP-042/ERNP-172 at a dose of 500 mg/kg, b.wt. respectively. All the rats received the respective assigned treatment for 14 days. On day 14, one hour after the respective assigned treatment all the rats, except group I, were administered with carbon tetrachloride at a dose of 0.5ml/kg, b.wt. in the form of equal mixture with liquid paraffin. In other words, it was given at the dose of lml/kg, b.wt. of carbon tetrachloride and liquid paraffin mixture (1:1 ratio). On 15^th day, 16 to 18 hrs post carbon tetrachloride administration, the blood was collected from the retro-orbital plexus under mild ether anesthesia and subjected for the estimation of markers of liver function such as SGPT and SGOT. The rats were sacrificed using deep ether anesthesia, Liver was weighed and small portion of liver was homogenized and subjected for estimation of glycogen.

Studv-2

Evaluation of different NP-042 extracts in an experimental model of thioacetamide- induced hepatic damage in rats:

Forty inbred male Wistar rats were divided in to 5 groups of 8 each. The rats from groups I and II received water once a day orally at a dose of 10 ml/kg b.wt. and served as normal and positive untreated controls respectively. Rats from groups III to V received NP-042, N P-042/A, NP-042/A/123, at a dose equivalent to lOOOmg/kg b.wt of extracts respectively. Ail the rats received the respective assigned treatment for 14 days. On day 14, one hour after the respective assigned treatment all the rats, except group I, were administered with hepatotoxin, thioacetamide 50 mg/kg b.wt. by subcutaneous route. On 15^th day, blood was collected from the retro-orbital plexus under mild ether anesthesia and subjected for the estimation of SGPT, SGOT and ALP.

Studv-3

Evaluation of N P-042 A and NP-042 A/123 extracts in an experimental model of carbon tetrachloride-induced hepatic damage in rats:

Forty-eight male Wistar rats were randomized to 6 groups of eight each. The rats of groups I and II received DM water once a day orally at a dose of 10 ml/kg b.wt. and were served as normal and untreated positive controls respectively. Rats from groups III to VI received NP-042/A (500mg/kg body wt. per day), N P-042/A ( lOOOmg/kg body wt. per day), NP-042/A/123 (500mg/kg body wt. per day) and N P-042/A/123 ( lOOOmg/kg body wt. per day) respectively. All the rats received the respective assigned treatment for 14 days. On day 14, one hour after the respective assigned treatment all the rats, except group I, were administered with carbon tetrachloride at a dose of 0.5ml/kg, b.wt. in the form of equal mixture with liquid paraffin. In other words, it was given at the dose of lml/kg, body weight of carbon tetrachloride and liquid paraffin mixture (1:1 ratio). On 15^th day, 16 to 18 hrs post carbon tetrachloride administration, the blood was collected from the retro-orbital plexus under mild ether anesthesia and subjected for the estimation of markers of liver function such as SG PT, SGOT and serum alkaline phosphatases. Statistical Analysis:

The Individual values were expressed as Mean ± SEM. The results were analyzed statistically using One-Way ANOVA to find out the level of significance followed by Dunnet's post comparison test. The minimum level of significance was fixed at p<0.05. The analysis was carried out using GraphPad Prism version 4.01. The V ranges from 7 to 8 in each group.

Results:

The first study (Study-1) was carried out to evaluate and compare the hepatoprotective potential of different N P-042/ERN P variants (NP-042/ERNP-167, N P-042/ERNP-168, N P- 042/ERNP-169, NP-042/ERNP-170, NP-042/ERN P-171 and N P-042/ERN P-172) in a well established experimental model of carbon tetrachloride-induced hepatic damage in rats. The extracts were tested at equal dose but at multiple doses (14 days). For the purpose of testing and comparing, the dose of extracts was fixed at a dose of 500mg/kg/day p. o, The carbon tetrachloride-intoxicated rats showed severe and significant derangement in liver function markers like enzymes SGPT and SGOT levels. Treatment with N P- 042/ERN P-171 significantly reversed the elevation of serum markers of hepatic damage like, SGPT, SGOT levels in carbon tetrachloride-intoxicated rats at both the tested doses. There is also a appreciable decrease in the mean values of the serum markers of liver function in NP-042/ERNP-170, NP-042/ERNP-171 and N P-042/ERN P-168 treated groups. From the results of above experiments, it can be inferred that, different N P- 042/ERN P extracts offered a appreciable hepatoprotection in rats against the hepatic damage induced by well known hepatotoxin carbon tetrachloride but to a varying extent, at selected dose, mode of administration and in method employed.

In the second set of study (Study-2), the different N P-042 extracts (NP-042, NP-042A and N P-042A/123) were evaluated/ compared for their hepatoprotective potential in experimental model of thioacetamide-induced hepatotoxicity in rats. The N P-042 extracts were tested at single dose level of dose level of 1000 mg/kg body weight/ day for 15 days. The thioacetamide-intoxicated rats showed severe and significant derangement in liver function markers like enzymes SGPT, SGOT and serum alkaline phosphatase (ALP). Treatment with NP-042/A/123 and N P-042/A significantly reversed the elevation of specific serum markers of hepatic damage like, SGPT, SGOT levels in rats with hepatic damage. The reductions in liver markers were also found to be reduced in other extract treated group (NP-042).

In third set of study (Study-3), the extracts N P-042/A and N P-042/A/123 were compared for their hepatoprotective potential in a well-known experimental model of carbon tetrachloride-induced hepatotoxicity in rats. Both the extracts were tested at two dose levels (500mg and 1000 mg/kg/day p.o. ). The assigned treatments were carried out for 14 days prior to challenge with a hepatotoxin, carbon tetrachloride. The carbon tetrachloride-intoxicated rats showed severe and significant derangement in liver function markers like enzymes SGPT, SGOT and also serum alkaline phosphatase levels. Treatment with NP-042A/123 significantly reversed the elevation of serum markers of hepatic damage like, SGPT, SGOT levels in carbon tetrachloride-intoxicated rats at both the tested doses. There is also a decrease in the mean values of the liver marker levels in NP-042A treated groups.

The following methodologies, examples, and tables were adopted for the various formulation of the herbal composition which illustrates the present invention:

Example 34

Total chemical constituents of various herbal blend extracts

Samples were analysed for total andrographolides, total tannins, flavonoids and alkaloids by in house specification methods. Results are mentioned in Table-2.

Table-2

Example 35

Standarisation of extracts by LCMSMS method analysis

LC-MS/MS optimization conditions: The LC-MS/MS analysis performed with Shimadzu LC-20 AD Prominence series HPLC system. LC separation was performed on a Reverse phase C18 (250 X4.6mm, 5um) Phenomenex column. The mobile phase consisted of 0.1% acetic acid in water and acetonitrile in a ratio of 90:10 (v/v), delivered at a flow rate of lml/min. The autosamp!er SIL-HTC (Shimadzu maker) injection volume was set at 20μΙ. UV detector chromatograms were monitor at 275nm and 254nm.

The API-2000 (Applied Biosystems, Canada) tandem triple quadrupole mass spectrometer equipped with an ESI source coupled with LC separation system in positive mode was used to ionize the molecules. Analyst 1.5 version was used for the control of equipment, acquisition and data analysis. ESI source conditions as follows: declustering potential (DP) 10 V, source gas (GS1) 50 psi and (GS2) 60 psi, curtain gas (CUR) 20psi, ion spray voltage (IS) 5500 V, focusing potential (FP) 400 psi and source temperature (TEM) 428 C were optimized with respect to ionization intensity response of the peak.

Sample solution preparation:

Individual sample solutions were prepared in the concentration of lmg/ml by weighing accurately 10 mg of individual samples in a separate 10 ml clean volumetric flask. The aqueous extracts were dissolved in water and remaining solvent extracts were dissolved in methanol by sonication for 10 min in an ultrasonic bath and finally made up to the volume respectively. The final sample solutions were filtered through 0.2μιτι syringe filter before loading to the autosampler.

The major chemical constituents detected in various herbal blend extracts were identified as fumaramine, cichoriin, chlorogenic acid, lactupicrin, protopine, wedeloactone, luteolin, andrographolide etc.

The results are mentioned in below Table-3

Name of herb in Chemical constituents MW (m/z) formulation identified

Cichorium intybus Lactucin 276.28

8-deoxylactucin 260

Chlorogenic acid 354.31

Cichoriin 340.28

Lactupicrin 410.13

Iso-chlorogenic acid 516.45

Fumaria indica d-hydrastine 383.39

Protopine 353.36

Cryptopine 369.41 d-bicuculline 367.35

Fumaramine 380.39

Ailanthus excelsa Glaucarubin 496.23

3,4-dihydroexcelsin 480.59

Glaucarubinone 494.53

Glaucarubol 396.43

Glaucarubolone 394.41

Eclipta prostrata Wedelolactone 314.24

Nor-wedelolactone 300.21

Beta-amyrin 426.71

Hentriacontanol 452.83

Luteolin 286.23

Andrographis 14-deoxy-ll, 12- 332.0 paniculata dihydroandrographoli

de

Neoandrographolide 480.0 14- 496.0

deoxyandrographolide

Panicolin 314.0

Homo- 344.0

androgdrapholide

14-deoxy- 12- 350.0

hydroxyandrographoli

de

14-deoxy-l l- 348.0

oxoandrographolide

14- 334.0

deoxyandrographolide

3,14- 318.0

dihydroxyandrograph

olide

Andrographiside 512.0

Andrographin 328.0

Andrographane 562.0

Example 36

Pharmaceutical formulations details

Direct compression of tablets

Procedure: Herbal actives and excipients were sieved through a 36 mesh sieve, resulting blend was then compressed into tablets. The composition of the tablets was as follows

Table-4

S.No. Ingredients Formula 1 Formula 2 Formula 3 Formula 4

(in mg) (in mg) (in mg) (in mg)

1 N P-042 SPD Extract 450 450 450 450

2 M icrocrystalline 200 375 170 270 cellulose 102

3 Maize starch 170 45 - -

4 Croscarmellose 35 - 25 - sodium 5 Magnesium stearate 5 5 - -

6 Sodium starch - 25 - - glycolate

7 Lactose DC - - 250 -

8 Colloidal Silicon - - 5 5 Dioxide

9 Dicalcium phosphate - - - 250 Dihydrate

10 PVP K 30 - - - 25

Total 850 900 900 1000

Results:

Direct compression tablets parameters are mentioned

Table-5

Granulation: By using Purified Water as a granulating fluid

PROCEDURE: Active and excipients were sieved through a 36 mesh sieve into a suitable mixer and granulated with a suitable quantity of boiled cooled purified water to form a medium /heavy granule. The granules were dried in a suitable oven at 60°C, until the LOD was <2%. The granulation was also tried with Isopropyl alcohol, alcohol etc.

BLENDING: The resulting dried granule was then passed through a 16 mesh sieve to give a granule and then blended with excipients. The resulting blend was then compressed into tablets using suitable capsule shaped tooling to give capsule shaped tablet, The composition of the tablets are mentioned in Table-6. Table -6

Formula Formula Formula Formula Formula Formula

SI.

Ingredients 1 2 3 4 5 6 No.

(in mg) (in mg) (in mg) (in mg) (in mg) (in mg)

NP-042 SPD

1 450 450 450 450 450 450 Extract

Microcrystalline

2 250 300 - - 300 150 cellulose

3 Maize starch - - - - 130

Talc - 50 - - - -

Croscarmellose

4 30 - 25 - 20 27 sodium

Magnesium

5 aluminum - - - - - 200 silicate

Pregelatinized

6 - - 80 - - - Starch

7 Lactose - - - 150 - -

Sodium starch

8 - 30 - 30 - - glycolate

Dicalcium

9 Phosphate - - 325 350 - -

Dihydrate

10 Purified water Qs Qs Qs Qs Qs Qs

B lending

Croscarmellose 18

11 15 - 15 15 15

sodium

Magnesium 5

12 5 5 - - 5

stearate

Sodium starch -

13 - 15 - - - glycolate

Colloidal Silicon -

14 - - 5 5 - Dioxide

Total 750 850 900 1000 920 850 RESULT: Wet Granulation by using excipients and Purified water as a granulating fluid, Results are mentioned in Table-7

Table-7

Capsule Formulation: By using purified water as a granulating fluid.

PROCEDURE: Active and excipients were sieved through a 36 mesh sieve into a suitable mixer and granulated with a suitable quantity of boiled cooled purified water to form a medium /heavy granule. The granules were dried in a suitable oven at 60°C, until the LOD was <2%.

BLENDING: The resulting dried granule was then passed through a 16 mesh sieve to give a granule and then blended with excipients. The resulting blend was then filled in suitable size capsules, the final composition of the capsules are mentioned in Table-8

Table-8

Formula 1 Formula 2

S.No. Ingredients

(in mg) (in mg)

1 NP-042 SPD Extract 450 450

Microcrystalline

2 200 350

cellulose

3 Maize starch 200 - 4 Croscarmellose sodium 27 30

5 Purified Water Qs Qs

B lending

6 Croscarmellose sodium 18 -

7 Magnesium stearate 5 5

8 Sodium starch glycolate - 15

Total 900 850

RESULT: Granules were filled in size '00' capsules (Table-9)

Table-9

Although this invention has been described by example and with reference to possible embodiment thereof, it is to be understood that modifications or improvements may be made thereto without departing from the scope of the invention.

Claims

CLAIMS We claim:

1. A herbal composition having hepato-protective activity comprising effective amounts of extracts of Eclipta prostrata, Cichorium intybus, Androgrophis paniculata, Fumaria indica, Ailanthus excelsa and suitable pharmaceutically acceptable carriers and additives.

2. The herbal composition as claimed in claim 1, wherein the extracts are obtained either by separate extraction or from the blend of Eclipta prostrata, Cichorium intybus, Androgrophis paniculata, Fumaria indica, Ailanthus excelsa parts.

3. The herbal composition as claimed in claims 1 & 2, wherein the extracts are obtained from the specific parts of the herbs preferably whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Androgrophis paniculata, whole parts of Fumaria indica and leaves of Ailanthus excelsa.

4. The herbal composition as claimed in claim 2 wherein the blend of herbs comprises whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Androgrophis paniculata, whole parts of Fumaria indica and leaves of Ailanthus excelsa in the ratio of 20:18:30:15: 17 respectively.

5. A herbal composition having hepato-protective activity comprising effective amou nts of extracts of Eclipta prostrata, Cichorium intybus, Androgrophis paniculata, Fumaria indica, Oxoxylum indicum and suitable pharmaceutically acceptable carriers and additives.

6. The herbal composition as claimed in claim 5, wherein the extracts are obtained either by separate extraction or from the blend of Eclipta prostrata, Cichorium intybus, Androgrophis paniculata, Fumaria indica, Oxoxylum indicum parts.

7. The herbal composition as claimed in claims 5 & 6, wherein the extracts are obtained from the specific parts of the herbs preferably whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Androgrophis paniculata, whole parts of Fumaria indica and leaves of Oxoxylum indicum.

8. The herbal composition as claimed in claim 6 wherein the blend of herbs comprises whole parts of Eclipta prostrata, seeds of Cichorium intybus, whole parts of Andrographis paniculata, whole parts of Fumaria indica and leaves of Oxoxylum indicum in the ratio of 20: 18:30: 15: 17 respectively.

9. The herbal composition as claimed in claims 1 & 5 wherein the preferable pharmaceutical carriers are Microcrystalline cellulose, Maize starch, Croscarmellose sodium, and mixtures thereof.

10. The herbal composition as claimed in claims 1 & 5 wherein the additives are in the form of blending agents, emulsifiers, preservatives, dispersants, and solvents known in the art.

11. The herbal composition as claimed in claims 1-7, wherein the said extracts are obtained by method selected from the group of percolation method, hot - soxlation method, super - critical - fluid method or enzyme extraction method .

12. The herbal composition as claimed in claims 1 - 7, wherein the said extracts are obtained by percolation method comprising the following steps of: a. Shade drying the material; b. Pulverizing the material obtained in step a. to coarse powder; c. Placing the powdered material in different percolators and extracting with n-hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1:1), methanol and water ( 1: 1) and water at room temperature for 24 h to 48 h; and d. Filtering the plant extracts and concentrating to dryness using a rotatory evaporator or on steam bath at optimum temperature under reduced pressure.

13. The herbal composition as claimed in claims 1 - 7, wherein the said extracts are obtained by hot-soxlation method comprising the steps of:

a. Placing the coarse powdered material in each soxlator using solvents n- hexane, acetone, ethyl alcohol, methanol, ethyl alcohol and water (1:1), methanol and water ( 1: 1) and water at refluxing temperature of each solvent;

b. recycling the process until extraction is completed; and

14. The herbal composition as claimed in claims 1 - 7, wherein the said extracts are obtained by super - critical - fluid extraction method comprising the steps of: a. Shade drying the material; b. Pulverizing the dried material to coarse powder; c. Placing the material in a SCF extractor at the temperature ranging from 40- 50 °C at high pressure range of 300-350 bar using carbon dioxide as super critical fluid for extraction upto 4 to 6 hours; and d. Collecting the extract in the collection vessel and evaporating at room temperature to remove any further residues of carbon dioxide.

15. The herbal composition as claimed in claims 1 - 7, wherein the said extracts are obtained by enzyme extraction method comprising the steps of: a. Shade drying the material; b. Pulverizing the dried material to coarse powder; c. Placing the material in Stainless Steel container and adding 0.5% to 5% of cellulase and pectinase enzyme combination in 4 volumes of distilled water; d. Processing the enzyme extraction at the temperature of 55°C to 60°C under occasional stirring upto 4-6 hours; and

e. Filtering the extract and concentrating the same to dryness on rotatory evaporator or on steam bath at optimum temperature under reduced pressure.

16. The herbal composition as claimed in claims 1 - 7, 11 and 15 wherein the yield from the enzyme extraction method is 8.99% for Eclipta prostrata, 12.45% for Cichorium intybus, 12.45% for Andrographis paniculata, 13.67% for Fumaria indica, 12.84% for Oxoxylum indicum, 13.5% for Ailanthus excelsa.

17. The herbal composition as claimed in claims 2, 4, 11 and 15 wherein the yield from the enzyme extraction method is 16.99% for herbal blend comprising Eclipta prostrata, Cichorium intybus, Andrographis paniculata, Fumaria indica and Ailanthus excelsa.

18. The herbal composition as claimed in claims 6, 8, 11 and 15 wherein the yield from the enzyme extraction method is 8.50% for herbal blend comprising Eclipta prostrata, Cichorium intybus, Andrographis paniculata, Fumaria indica and Oxoxylum indicum.

19. The herbal composition as claimed in claims 1 - 18 wherein the composition is in the form of a tablet, capsule, syrup, injection or any other suitable form.