DESC:FIELD OF THE INVENTION
The present invention relates to a herbal composition and use for treating patients with liver diseases. The present invention also relates to a method for making the herbal composition and methods for treating patients with the said composition.

BACKGROUND OF THE INVENTION
In recent years, westernization of eating habits has progressed, and lifestyle-related diseases such as diabetes, hyperlipidemia, fatty liver, obesity and arteriosclerosis are increasing. These increased incidences are not genetic but mainly due to environmental factors. For example, abnormal lipid metabolism due to intake of high-fat and high-calorie foods causes blood lipid elevation, development of insulin resistance, adipocyte hypertrophy, insulin secretion failure, and the like. As a result, diabetes, obesity, arteriosclerosis, etc. occur frequently, leading to progression of the disease state.

Fatty liver is a disease in which triglycerides accumulate in the liver due to excessive intake of fat and alcohol and abnormal lipid metabolism in the liver. Clinically, when the fat is observed in 5% or more of hepatocytes or when the fat is more than 5 mg per 100 mg of liver, it is classified as fatty liver. In recent years, patients with fatty liver have been increasing rapidly due to high fat, high calorie diet and lack of exercise due to the development of civilization, and the age group has been on the whole from the teens to the elderly. When left untreated, liver can be progressed to hepatitis, liver cirrhosis, liver cancer and the like.

There are two main types of fatty liver disease: non-alcoholic and alcoholic. The former is not related to heavy alcohol use and results from fat accumulation in the liver with little to no inflammation or liver cell damage. This is called simple fatty liver and generally doesn’t progress to a point of liver damage. Fatty liver can also develop in non-heavy drinkers with concurrent inflammation and liver cell damage; this condition is known as non-alcoholic steatohepatitis, and is associated with progressive scarring of the liver (cirrhosis) and liver cancer.

Alcoholic steatohepatitis is caused by alcohol abuse and only happens in people who are heavy drinkers and those who have been drinking for a long period of time. Alcoholic fatty liver disease is usually a silent disease with few or no symptoms. The person may feel tired or have some aches in the upper right side of the abdomen. This is the earliest stage of alcohol-related liver disease. More severe or progressed forms of alcoholic liver disease include alcoholic hepatitis and cirrhosis.

For the treatment of fatty liver, fibrates such as clofibrate and bezafibrate are prescribed in parallel with exercise therapy and dietary restrictions. Its mechanism of action is by inhibiting the synthesis of triglycerides in the liver. In addition, nicotinic acid preparations are drugs with a similar mechanism of action. On the other hand, foods for specified health use, foods containing globin proteolysates and oolong tea polyphenols that suppress the absorption of ingested neutral fat are on the market.

Rhabdomyolysis is a side effect of fibrate drugs. This is a condition in which a part of skeletal muscle is degenerated/ necrotic, and myoglobin flows into the blood. The outflow of muscle components places a load on the renal tubules and is often associated with acute renal failure. In addition, there is stomach discomfort and skin rashes, and special attention is required especially for patients with hepatic/ renal dysfunction and diabetic patients. The nicotinic acid preparations are also likely to cause stomach discomfort, diarrhoea, rash, flushing of the face, headache, warmth, and sweating palpitations.

There are some patent documents that describe the use of herbal compositions for treatment of fatty liver.
KR100604354B1 discloses a pharmaceutical composition for the prevention and treatment of liver diseases containing herbal extracts, and more particularly to a pharmaceutical composition for the prevention and treatment of liver diseases, which essentially contain phosphorus, Baekchul, Fingering, Seungsong and Nyo extract.

KR101523663B1 relates to a pharmaceutical composition for preventing or treating fatty liver disease or obesity, which comprises a mixed herbal extract of Arctium lappa Linne, Glycyrrhiza uralensis Fischer, Ginger (Zingiberis rhizoma Crudus) and Magnoliae Cortex.

CN105396103A relates to a preparation for rapid treatment of fatty liver which is prepared from the following herbal raw materials: Abrus cantoniensis, Semen Sinapis, Allium macrostemon, Filiform cassytha herb, Elsholtzia blanda, Asparagus cochinchinensis, Radix angelicae pubescentis, Phyllanthus urinaria, Hedyotis diffusa, Scutellaria baicalensis, Ginger, Elsholtzia ciliata, Fritillaria thunbergii, Polygala japonica, Seaweed, Chinese Buckeye Seed, Chicken's gizzard-membrane, Amomum kravanh, Poncirus trifoliate, Cortex Lycii, Curcuma zedoaria, Ruddle, Dragon bone, and Swertia bimaculata.

CN105999209A discloses a traditional Chinese medicine prepared from, by weight, 2-10 parts of Ginger, 16-22 parts of Lily, 10-15 parts of Fructus gardeniae, 15-22 parts of Ganoderma lucidum, 5-8 parts of Radix scutellariae, 10-15 parts of Wrinkled gianthyssop herb, 7-10 parts of Semen cassiae and 2-7 parts of Calculus bovis.

CN102198168A relates to a product made from Rosemary extract for treating alcoholic livers and fatty livers, and a processing technology of the product. According to the invention, fat-soluble extracts and water-soluble extracts of Rosemary are processed through an ultra-low temperature extraction technology, and are formulated and combined into finished products.
KR101160088B1 discloses a composition containing a mixed extract of Prickly pear, Brown root, Licorice, Cabbage, Baekchul, Baekbaekpi, Ginseng for inhibiting alcoholic fatty liver and hyperlipidemia and hangover.

KR20150118774A relates to a pharmaceutical composition comprising an extract of Angelica gigas and Zygotes, Angelica gigas, Radix ginseng, and Gardenia for prevention or treatment of fatty liver.

US9962420B2 discloses compositions formulated in the form of tablets, wafer capsules, gel capsules, sticks, sachets, vials, droppers or in injectable form. The compositions comprise a mixture of: an extract of Chrysanthellum indicum, an extract of Cynara scolymus, an extract of Vaccinium myrtillus, an extract of Olea europaea, and pipeline.

Although the prior art discloses compositions comprising different combinations of herbs or plant extracts for the treatment of fatty liver, there is still a need for a composition of readily available herbal extracts that acts on multiple targets, has fewer side effects and is economical.

OBJECTS OF THE INVENTION
The main object of the present invention is to provide a synergistic composition of herbal extracts for treating patients with liver diseases.

Another object of the present invention is to provide a composition comprising a mixture of extracts from Piper longum, Nigella sativa, Eclipta alba, Zingiber officinale and Cinnamomum zeylanicum.

Yet another object of the present invention is to provide a composition that is a mixture of active components piperine, thymoquinone, wedelolactone, 6-shogaol and cinnamaldehyde.

Still another object of the present invention is to provide a method for making the herbal composition.

Further object of the invention is to use the synergistic composition of herbal extracts for treating patients with liver diseases.

SUMMARY OF THE INVENTION
The herbal composition of present invention comprises a mixture of extracts from Piper longum, Nigella sativa, Eclipta alba, Zingiber officinale and Cinnamomum zeylanicum.

In an embodiment the herbal composition of present invention comprises a mixture of 2.5 – 4mg Piper longum extract, 26.5 – 43.75mg Nigella sativa extract, 126.25 – 210mg Eclipta alba extract, 112.5 – 150mg Zingiber officinale extract and 11.25 – 15mg Cinnamomum zeylanicum extract.

In another embodiment, the herbal composition of present invention is a mixture of active components - piperine, thymoquinone, wedelolactone, 6-shogaol and cinnamaldehyde.

In another embodiment the present invention also discloses the method for preparing the composition of herbal extracts.

In still another embodiment the present invention provides the use of the herbal composition for the treatment of liver diseases mainly alcoholic fatty liver and its associated complications.

DETAILED DESCRIPTION OF THE INVENTION
The inventors of the present invention have taken intensive efforts to develop a composition of herbal extracts that has lower side effects while effectively treating liver diseases mainly alcoholic fatty liver and its associated complications.

The composition of the present invention has a unique combination of herbal extracts scientifically proven to be effective in different stages of the disease as it comprises of maximum concentration of active principles with specifications to its mechanism of action.

In an embodiment, the present invention provides a synergistic composition comprising a mixture of extracts from Piper longum, Nigella sativa, Eclipta alba, Zingiber officinale and Cinnamomum zeylanicum.

Piper longum (Long pepper), sometimes called Indian long pepper or pipli, is a flowering vine in the family Piperaceae. It is cultivated for its fruit, which is usually dried and used as a spice and seasoning. Ayurveda has explained various uses of long pepper for dietary purpose as well as for various health purposes. It is basically used for healthy respiratory system. Piper longum is also beneficial for healthy digestion and healthy metabolism.

Nigella sativa (Black cumin), also called black seed, black caraway, Roman coriander, kalonji, or fennel flower, annual plant of the Ranunculus family (Ranunculaceae), grown for its pungent seeds, which are used as a spice and in herbal medicine. Nigella sativa seeds contain a high amount of thymoquinone, the main active within the seeds, which is a powerful antioxidant. Black seed oil has shown promising results in treating some of the most common health conditions, including high blood pressure and asthma. Black seed oil is also thought to have anticancer properties.

Eclipta alba commonly known as False daisy, Karisalankanni, and Bhringraj, is a species of plant in the sunflower family. The plant has diverse medicinal values and is commonly used for treatment of gastrointestinal disorders, respiratory tract disorders (including asthma), fever, hair loss and graying of hair, liver disorders (including jaundice), skin disorders, spleen enlargement, and cuts and wounds.

Zingiber officinale (Ginger) is a rootstock that grows sideways and is fleshy, lumpy, yellowish, with a spicy and fragrant smell. It is a rhizome of a plant and dried roots are used as medicine. It has been known to treat chills, fever, headache, vomiting, seawater and sputum caused by a cold, as well as diarrhea caused by food poisoning.

Cinnamomum zeylanicum (Ceylon cinnamon, true cinnamon) has been prized for its medicinal properties for thousands of years. Cinnamon, obtained from inner bark of Cinnamomum trees, is a spice with distinct smell and flavor due to the oily part, which is very high in the compound cinnamaldehyde. Cinnamon contains large amounts of highly potent polyphenol antioxidants. The antioxidants in cinnamon have anti-inflammatory effects, which may help lower the risk of disease. Cinnamon may improve some key risk factors for heart disease, including cholesterol, triglycerides and blood pressure.

In a preferred embodiment, the synergistic composition of the present invention comprises a mixture of following active ingredients obtained from the described herbal extracts - piperine, thymoquinone, wedelolactone, 6- shogaol and cinnamaldehyde.

In an embodiment, the composition of the present invention comprises a mixture of 2.5 – 4mg Piper longum extract, 26.5 – 43.75mg Nigella sativa extract, 126.25 – 210mg Eclipta alba extract, 112.5 – 150mg Zingiber officinale extract and 11.25 – 15mg Cinnamomum zeylanicum extract.

The herbal extracts described in the present invention, may be obtained via any suitable process known in the art, in the form of powder.

The composition of the present invention may further comprise suitable excipients. The amount of excipient used depends on the type of formulation, and one type of excipient can perform more than one function.

The excipient as used herein may be selected from diluent, lubricant, disintegrant, suspending agent and preservative.

Diluents are inert excipients that are used to adjust the bulk in a composition. Commonly used diluents comprise of lactose, dicalcium phosphate, micro crystalline cellulose, kaolin, mannitol and starch.

Lubricants are used in the composition to prevent adhesion of the powder. Commonly used lubricants comprise of magnesium stearate, calcium stearate, talc, stearic acid and polyethylene glycol.

Disintegration includes at least one of alginic, carboxymethylcellulose, clays, croscarmellose sodium, sodium alginate and cross-linked polyvinylpyrrolidone.

Suitable preservatives include, but are not limited to, benzyalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, sodium benzoate and the like.

If desired, flavoring, coloring and/or sweetening agents may be added as well.

Said herbal composition of the present invention may exist in various formulations. These formulations include, but are not limited to, liquid, semi-solid and solid pharmaceutical formulation, such as dispersion or suspension, tablet, capsule, pellet, granules and powder. The preferred oral formulation of the herbal extracts according to the present invention include capsule.

In an embodiment the herbal composition is prepared by sifting the herbal extracts through size 40 mesh and adding the excipients to get the final formulation.

In further embodiment, the composition of herbal extracts of the present invention is used for treating and/or preventing liver diseases, including, but not limited to alcoholic or fatty liver, liver cirrhosis and viral hepatitis. The patients with liver disease are administered a therapeutically effective amount of the herbal extract composition of the present invention.

In preferred embodiment, the herbal composition of present invention is used to treat alcoholic fatty liver disease.

The synergistic composition of herbal extracts of the present invention contains potent "liver specific" active ingredients that act on different stages of the alcoholic liver disease.

In an embodiment the herbal composition of the present invention shows anti-inflammatory action due to inhibition of prostaglandin release.

Piperine is the main compound present in black pepper extract and it has demonstrated remarkable antioxidant, antitumor, and drug availability-enhancing characteristics of this compound, together with immunomodulatory potential. Piperine inhibits the production of prostaglandin E2 (PGE2) and nitric oxide induced by lipopolysaccharide while decreasing tumor necrosis factor-a (TNF-a), inducible NO synthase (iNOS) and cyclooxigenase-2 (COX-2) thereby resulting in anti-inflammatory activity.

The Nigella sativa extract contains the active ingredient thymoquinone, is a promising agent to improve hepatic steatosis, oxidative stress; inflammatory, apoptotic status, fibrosis and so prevent liver damage.

The Eclipta alba extract used in the present invention contains the active constituent, wedelolactone, which up-regulates the protein levels of adenosine monophosphate activated protein kinase (AMPK) and peroxisome proliferator-activated receptor-alpha (PPARa) as well as the gene expression of AMPK, PPARa, lipoprotein lipase (LPL), and the low-density lipoprotein receptor (LDLR). Furthermore, wedelolactone also increases the activities of superoxidase dismutase (SOD) and glutathione peroxidase (GSH-Px) and decreases the level of the lipid peroxidation product malondialdehyde (MDA) in the liver, therefore decreasing the activity of alanine aminotransferase (ALT). Thus, wedelolactone possesses lipid-lowering and steatosis-improving effects.

The Zingiber officinale extract used in the present invention, contains the main constituent 6-shogaol, which possess beneficial health properties such as anti-inflammatory and chemo preventive effects. The 6-shagaol decreases the diethyl nitrosamine (DEN)-mediated elevations of serum aspartate transaminase and alanine transaminase as well as the DEN-induced hepatic lipid peroxidation, enhancing the antioxidant defense mechanism.

The Cinnamomum zeylanicum extract used in the present invention contains the active ingredient, cinnamaldehyde, which increases peroxisome proliferator-activated receptors (PPARs) transcription. Cinnamaldehyde induces the expression of PPARd and PPAR? target genes, namely aP2 and CD36, in differentiated adipocytes. As a result, PPARd, PPAR? and their heterodimeric partner RXR appear to play a part in the cinnamaldehyde action in the target tissues, thereby enhancing insulin sensitivity and fatty acid ß-oxidation and energy uncoupling in skeletal muscle and adipose tissue.

Also, the inventors of the present invention believe that the active compounds of the herbal composition of the present invention can enhance the antioxidant defense systems, such as glutathione peroxidase and glutathione S-transferase, and reduce levels of malondialdehyde (MDA) and hepatic steatosis. The main reason for the protective feature is the effect of the active compounds on the expres¬sion of proliferating cell nuclear antigen (PCNA). PCNA is a nuclear protein that is involved in regulating cell pro¬liferation. In liver disorders the expression of PCNA is increased, which ultimately can result in excessive and uncontrolled proliferation of tissue to replace the damaged tissue and causes liver fibrosis pro¬gression.

From the details presented in the disclosure it is evident that the composition of the present invention is not a mere admixture resulting in a composition having the aggregation of the properties of the components used but a composition formed by the synergistic activities of the components used.

The following examples are for the purpose of illustration of the invention only and are not intended in any way to limit the scope of the present invention.

EXAMPLES OF THE INVENTION

No Ingredients % w/w
1 Piper longum extract 0.877
2 Nigella sativa extract 9.294
3 Eclipta alba extract 44.276
4 Zingiber officinale extract 39.454
5 Cinnamomum zeylanicum extract 3.945
6 Di-calcium Phosphate 1.754
7 Bronopol 0.200
8 Magnesium Stearate 0.200
Average Weight 100%

The herbal extracts are sifted through Size 40 mesh and mixed in a blender for 10 min. Magnesium stearate, Di-calcium phosphate and Bronopol are added to the blender and mixing continued for 5 min. The powder obtained is filled into a Size 2 capsule. The Empty Capsule Weight was 63 mg, Weight of Net content was 285.140 mg and the Average weight of Capsule was 348.140 mg.
The capsule formulation was tested for Microbial content and the results are:

Product Dilution Bacterial Result Bacterial Limit Fungal Result Fungal Limit
Herbal extracts Capsule 10-2 650 cfu/g 1000 cfu/g <10 cfu/g 100 cfu/g

The capsules prepared by the process of present invention were evaluated of toxicity and hepatoprotective activity against chronic liver toxicity in Wistar rats. The rats were administered the test substance orally by dissolving the capsules in distilled water. For the study adult male albino Wistar rats (150 ± 20g) were divided into 6 groups. The group wise administration was done for 25 days –
Group 1: saline (1 ml p.o), Group 2: ethanol (3.76 g/kg) + silymarin (100 mg/kg) simultaneously, Group 3: ethanol (3.76 g/kg) + test capsules (100 mg/kg) simultaneously, Group 4: ethanol (3.76 g/kg) + test capsules (200 mg/kg) simultaneously, Group 5: ethanol (3.76 g/kg) + test capsules (400 mg/kg) simultaneously and Group 6: ethanol (3.76 g/kg).
The blood sample were collected on Day 1 and Day 26 and were centrifuged at 3000rpm for 30min at 4°C for 20mins to obtain the serum for analysis of various biochemical parameters. Determination of liver enzymatic markers (SGOT, SGPT, ALT, AST, ALP), non- enzymatic markers (cholesterol and triglycerides), total protein levels were estimated according to the commercial kit manufacturer’s instructions (Human Gesellschaft, Germany).

TABLE 1: DETERMINATION OF LIVER ENZYMATIC PARAMETERS

S.NO GROUP SGOT (U/L) SGPT(U/L) ALP(U/L) AST(U/L)
1 I (Normal) 190.60 ± 0.12 97.97 ± 0.12 92.87 ± 0.12 37.4 ± 3.1
2 II (Silymarin) 152.93 ± 0.09* 76.59 ± 0.10* 53.03 ± 0.23* 45.6± 2.5*
3 III (100 mg/kg) 185.14 ± 0.05 89.28 ± 0.02 69.84 ± 0.09 143.4 ± 2.5*
4 IV (200 mg/kg) 154.33 ± 0.02* 79.40 ± 0.04* 61.19 ± 0.03* 119.4 ± 4.6*
5 V (400 mg/kg) 207.06 ± 0.11 105.67 ± 0.01 73.79 ± 0.08 92.7 ± 3.1*
6 VI (Alcohol) 240.70 ± 0.06 160.98 ± 0.05 105.18 ± 0.11 195.5 ± 5.6

Values are expressed as Mean ± SD of 6 individuals.
*P<0.01, compared with Alcohol control

TABLE 2: DETERMINATION OF LIVER NON-ENZYMATIC PARAMETERS
S.NO GROUP CHOLESTEROL TG (mg/dl) HDL
(mg/dl) VLDL
(mg/dl) LDL
(mg/dl)
1 I (Normal) 68.16 ± 0.12 580.63 ±
0.11 30.65 ±
0.09 116.10 ±
0.18 30.30±
0.24
2 II (Silymarin) 80.11 ± 0.21* 345.27 ±
0.03* 26.01 ±
0.12* 69.03 ±
0.06* 570±
0.38*
3 III (100
mg/kg) 55.63 ± 0.16 698.50 ±
0.07 28.20 ±
0.04 86.24 ±
0.14* 26.54±
0.11
4 IV (200
mg/kg) 66.69 ± 0.11 721.87 ±
0.04 32.29 ±
0.07* 144.33 ±
0.09 31.75±
0.24*
5 V (400
mg/kg) 70.85 ± 0.19 564.62 ±
0.10* 52.19 ±
0.01 112.87 ±
0.02* 29.30±
0.27*
6 VI (Alcohol) 61.52 ± 0.15 658.35 ±
0.05 38.39 ±
0.04 131.60 ±
0.20 32.75 ±
0.31

Values are expressed as Mean ± SD of 6 individuals.
*P<0.01, compared with Alcohol control

TABLE 3: DETERMINATION OF TOTAL PROTEIN LEVELS

S.NO GROUP TOTAL PROTEINS (mg/dl)
1 I (Normal) 8.70 ± 0.14*
2 II (Silymarin) 8.20 ± 0.08*
3 III (100 mg/kg) 8.53 ± 0.02*
4 IV (200 mg/kg) 7.83 ± 0.09*
5 V (400 mg/kg) 7.90 ± 0.11*
6 VI (Alcohol) 10.07 ± 0.06*
As per the study outcome the test capsules, obtained from the present invention, showed decreasing serum levels of enzymatic (ALT and AST) transaminase, alkaline phosphatase, SGOT, SGPT, non-enzymatic parameters such as triglycerides and total cholesterol. Also, the test capsules, disclosed in the specification, improved the serum protein levels around the normal values after the decrease that was seen in the alcohol treated groups. The capsules from present invention also prevented histopathological changes in the liver, kidneys and lungs due to the toxicity of ethanol.
Further the test capsules, obtained from the disclosure of present invention, were also assessed for the toxic potential, when administered by oral gavage, in a single dose to Wistar rats. The test capsules were administered as a single dose at an equivolume of 1ml/100g body weight for all animals and the actual volume if administration was calculated based on the most recent body weight of animals. Observations were made including changes in skin and fur, eyes, mucous membranes, tremors, convulsions, salivation, diarrhea, sleep and coma and also respiratory, autonomic, somatomotor activity and behavioral pattern in the animals. The body weight of each rat was recorded prior to treatment on Day 1, then weekly and at the terminal sacrifice time (Day 14).

The test capsules did not reveal any clinical signs, mortalities and gross pathological changes when administered once by oral gavage route in Wistar rats at a dose of 2000mg/kg. Hence, it can be concluded that test capsules, prepared by the present invention, are safe and non-toxic in Wistar rats up to a dose of 2000mg/kg.

An open label prospective clinical study to evaluate the efficacy of the capsules, obtained from the present invention, for alcoholic liver disease is underway. The objective of the study is to evaluate safety and efficacy compared to standard of care for alcoholic fatty liver. The tentative dose is in the range of 50-2000 mg/kg. Exact dose will be derived from the clinical study.
,CLAIMS:We claim,
1) A herbal composition comprising a mixture of extracts from Piper longum, Nigella sativa, Eclipta alba, Zingiber officinale and Cinnamomum zeylanicum.

2) The herbal composition according to claim 1, comprises a mixture of 2.5 – 4mg Piper longum extract, 26.5 – 43.75mg Nigella sativa extract, 126.25 – 210mg Eclipta alba extract, 112.5 – 150mg Zingiber officinale extract and 11.25 – 15mg Cinnamomum zeylanicum extract.

3) The herbal composition according to claim 1 and 2, is a mixture of active components piperine, thymoquinone, wedelolactone, 6-shogaol and cinnamaldehyde.

4) The herbal composition of the preceding claims, wherein the herbal extracts are obtained as powder via any suitable process known in the art.

5) The herbal composition of the preceding claims, may be formulated as liquid, semi-solid and solid form

6) The herbal composition of preceding claims, preferably is formulated as dispersion or suspension, tablet, capsule, pellet, granules or powder dosage form.

7) The herbal composition of earlier claims, is most preferably formulated as capsule dosage form.

8) The herbal composition of preceding claims, is used for treating and/or preventing liver diseases, including, but not limited to alcoholic or fatty liver, liver cirrhosis and viral hepatitis.

9) The herbal composition as described in claims of present invention, is used for the treatment of alcoholic fatty liver disease.

10) The herbal composition of preceding claims, is prepared by sifting the herbal extracts through Size 40 mesh, mixing in blender for 10min and formulating the powder in desired dosage form.