WO2003030635A1

WO2003030635A1 - A herbal hepatoprotective and weight gain promoter and a process thereof

Info

Publication number: WO2003030635A1
Application number: PCT/IN2001/000178
Authority: WO
Inventors: Ravindra Kumar Agarwal; Amit Agarwal
Original assignee: Natural Remedies Private Limited
Priority date: 2001-10-11
Filing date: 2001-10-11
Publication date: 2003-04-17

Abstract

The present invention relating to herbal hepatoprotective and weight gain promoter which comprises the synergistic mixture of whole plant of Andrographis paniculata, Solanum nigrum, Phyllanthus amarus and Boerhaavia diffusa. The present invention also relates to a process of preparation of such composition. The composition is particularly useful as hepatoprotective and weight gain promoter.

Description

"AHERBAL HEPATOPROTECTIVE AND WEIGHT GAIN

PROMOTERAND A PROCESS THEREOF"

The present invention relates to a novel herbal composition useful for hepatoprotection and promoting weight gain in domestic animals. One embodiment of the present invention resides in providing a herbal composition which is synergistically effective in promoting weight gain in domestic animals such as poultry, cattle etc through optimization of liver function. The composition of the present invention is also a very potent and effective liver tonic. The composition guards against toxins and toxic effects of feed contaminants and several chemical drugs. It helps to regenerate damaged liver tissues caused by infections, toxins and exhibits hepatogenic property through optimization of liver function. The composition is unique because of its multifunctional properties as well as its activity through optimization of liver functions. An herbal composition having such multifunctional properties and activity through optimization of liver functions to promote weight gain in domestic animals is hitherto not known either in modern medicine or other hitherto marketed herbal products. The composition is equally useful in case of Aflatoxicosis and also improper fat metabolism in domestic animals. The composition is a choleretic and thus it normalises fat metabolism and in turn causes improvement in poor feed conversions.

Livestock suffer from drop in optimum functioning of liver, caused by toxins, chemical drugs, certain feed contaminants, parasitism, infections etc. The present invention serves as a highly efficient liver tonic, rectifies deranged liver and optimizes all liver functions. Similarly poultry birds so susceptible to above factors and often suffer from liver problems.

Liver is considered to be one of the most important organs of human beings and animals. It is responsible for maintaining the body^'s metabolic homeostasis. The liver serves many functions, including regulation of blood volume and manufacture of certain blood clotting factors, the storage of several substances, including glycogen, copper, iron and Vitamin Bι₂; the metabolism of proteins, carbohydrates, and fats; production of heat; detoxification (removing the poisonous effect) of certain foreign substances in the blood; destruction of old red blood cells; and formation of bile. Specialized functions of the liver that can be emphasized include processing of dietary amino acids, carbohydrates, lipids and vitamins, phagoc>1osis of particulate material in the splanchnic circulation, synthesis of serum proteins, bio-transformation of circulating metabolites and detoxification and excretion into bile of endogenous waste products. The variety of functions performed by the liver makes it vulnerable to a wide variety of metabolic, toxic, microbial. circulatory and neoplastic related diseases. Primary diseases of liver include jaundice, hepatitis and cirrhosis of the liver. Secondary diseases of the liver are generally the damages caused in cases of l alcoholism, cardiac decomposition, extrahepatic infections and the like. Consequences of a deranged liver are generally life threatening and often fatal.

Digestion is a very important and vital function carried out by the body both in animals and humans which serves as an energy generator to run the body mechanisms. Improper digestion apart from causing discomfiture also results in incomplete absorption and assimilation of nutrients and hence poor feed conversion. The poor feed conversion in turn affects general health of the domestic animals such poultry, cattle, birds etc as in the case of humans. Consequently, it results in reduced productivity, livability and hinders uniform sexual maturity. To counteract these effects and to positively promote growth, antibiotics have been in use for many years. Table- 1 lists the antibacterial growth promoters and toxin-binders available in the market.

TABLE-1

ANTIBACTERIAL GROWTH PROMOTERS

_>duct Active ingredients Mixing Rate

1. Zinc bacitracin 100-200 gms/tonne 2. Oxytetracycline 100-500gms/tonne Vergimamycin_^ 500 gms/tonne

4. FlavophosjΛolijrøl 100 gms/tonne TOXIN B INDERS

1. Organic acids, Gentian voi let and zeolites 2.5kgs-10kgs/tonne depending on moisture

2. Phytosilicates, Zeolites 500gms-2.5 kg tonne of feed

3. Zeolites, organic acids and probiotics 1.5 kgs/tonne of feed for moisture level upto 15%

However, treatment using antibiotics has its merits and demerits. Its merits are: l .They may inhibit the growth of microorganisms that produce excessive amounts of Ammonia and other toxic nitrogenous waste products in the intestine. The intestinal bacteria are altered in such a way that less urease is produced and thus lesser Ammonia is generated. Ammonia is a highly toxic gas (permissible level only 25 ppm) and suppresses the growth of birds.

2.They may improve the availability or absorption of certain nutrients like Calcium, Phosphorus. Magnesium, protein, amino acids and vitamins.

3. They eliminate pathogens causing sub-clinical infection. In many instances, they may actually cure pathological diseases (mainly confined to the intestinal tract) and control low disease levels which may be encountered even in good, healthy poultry. 4.They destroy the microbial flora causing inflammation (therefore swelling and thickening of the intestinal wall), and thus help to maintain thinner intestinal wall and thereby effect better absorption of nutrients.

5.Antibiotics improve feed or water consumption or both. General health of the bird and feed conversion improves. It is reported that ceca of antibiotic fed chicken are larger and filled with greater quantity of moist excreta, compared to the ceca of chickens fed the same diet without an antibiotic.

Reference in this connection may be made to "Nutrition of Chicken", Ed. M.L. Scott, M.C. Nesheim and R.J. Young, Ithaka, New York, 1982, 388-390; "Feeds & Principles of Animal Nutrition", Ed. G.C. Banerjee, Oxford & IBH Publishing Co. Pvt. Ltd., Calcutta, 1988, 375.

The demerits of using antibiotics are highlighted below:

1.Antibiotics are generally non-specific and may also destroy the beneficial organisms in the Gastro Intestinal Tract (GIT).

2.Having destroyed or impaired the beneficial organisms, absorption of nutrients ih the GIT is adversely affected even though the nutrients are present in adequate amounts in the diet, their absorption is hindered and thus is lost to the birds.

3. It is feared that widespread use of antibiotics in food animals can lead to development of resistance in humans.

A classic example is that of Avoparcin. Avoparcin was a well-known glycopetode antibiotic. which was used in many countries as a growth enhancer in various species of livestock including pigs and poultry. In 1995, this product was banned in Denmark and in Germany in 1996. It was banned for use in European Union in 1997. This was because of concerns that the use of Avoparcin was leading to the development of resistance against an important human medicine, Vancomycin and that the resistance could transfer from bacteria in the food chain to man. This could be important as Vancomycin is a so called 'antibiotic of last resort' in human medicine.

Reference in this context may be made to International hatchery practice, 1998Nol.12, No.8. Published by. P.A.P. U.K

4. Some antibiotics are suspected to be carcinogenic. Like for instance, Nitrofurans have been removed from the Canadian market as of February 1^st 1993. Concerns about the carcinogenicity of Nitrofurazone in rats and mice based on US data promoted Canadian action. The Branch subsequently ruled that the sale of the feed supplement (drugs) for use in food-producing animals should cease.

Reference in this connection may be made to Animal Pharm, No. 270, February 12^th 1993, 13 5. Another major drawback with the use of certain antibiotics is that they are also absorbed in GIT of the bird, along with other nutrients. In such cases a specific withdrawal period is required, during which these absorbed antibiotics are dislodged or metabolized and are not retained in the body in their original form. In case this withdrawal time is not adhered to strictly before slaughter (as is often the case) the bird meat retains the antibiotic(s) and can lead to problems if the bird meat is used for human consumption. Table-2 lists the withdrawal time and limitations for use for certain antibiotics.

TABLE-2

DRUG PRES AUGHTER LIMITATIONS FOR USE TOLERANCE LEVEL

WITHDRAWAL

TIME (PPM)

Aklomide,roxarsone 5 Do not use in egg-laying birds; Aklomide: liver, use as only source of organic (Aklomix-3) muscle,4.5;skin,fat,3.Arseni arsenic c:muscle,eggs,0.5; other by- roducts,2

Aklomide,sulfanitran, 5 Do not use in egg-laying birds; Aklomide and arsenic same roxarsone(No vastat- 3 ) use as only source of organic as above; sulfanitran,0 arsenic

Amprolium 0 For control only several Ei-meria Liver,kidney, 1 ; sp. muscle,0.5;whole eggs,4;egg yoiks,8 Arsenic, same as above

Arsanilic acid or 5 Use as only source of organic Arsenic, same as above sodium arsanilate arsenic

Bacitracin 0 Meat, eggs,0.05(0.02 unit/g)

Buquinolate 5 Use as only source of organic Liver, arsenic kidney,skin,0.4;muscle,0.1 ; eggs,0.2(calculated as arsenic)

Butynorate,sulfanitan, 5 Do not use in egg-laying birds; Butynorate and dinsed,none dinsed,roxarsone(Poly use as only source of organic published;aresenic,same as stat) _ _ arsenic above

Carbomycin 2 Do not use in egg-laying birds; 0

Chlortetracycline 1 Do not use in egg-laying birds Kidney,4; when fed at 500g/900 kg feed muscle,liver,fat.skin, 1 ;eggs. 0

Clopidol(Coyden) Do not use in chickens over 16 Liver and weeks old when feeding 0.025%, kidney, 15;muscle,5 or reduce dosage to 0.0125% 5 days before slaughter

Decoquinate 5 Do not use in egg-laying birds Muscle, 1 ; other tissues,2

Erythromycin(Gallim 1-2 Do not use in egg-laying birds at Eggs,0.025,edible ycin) a level over 185g 900 kg feed tissues.0.125

Estradiol 42 Injected subcutaneously at base ofO monopalmitate skull at not less than 5 weeks of

.age_. _ ^'

Furazolidone (Furox 5 Do not use in egg-laying birds or None published NF-180) in replacements over 14 weeks old Gentamicin sulfate 35 For use in day-old chickens only None published injection

Hygromycin B 0 8-12 g/900 kg feed 0

Lasalocid sodium 8-12 g/900 kg feed Edible tissues,0.05

Lincomycin 8-12 g/900 kg feed Edible tissues,0.1

Metoserpate Do not use in egg-laying birds 0.02 hydrochloride

(Pacitron)

Monensin 3-5 Do not use in egg-laying birds 0.05 sodium(Coban)

Nequinate(Statyl) Use as only source of organic Edible tissues,0.1 arsenic; do not use in chickens over 16 weeks old

Nicarbazin (Nicarb) 4 ?_ not use in egg-laying birds 4

Nitarsone(Histostat- 5 Use as only source of organic None published 501 ___ arsenic

Nitrofurazone(NFZ,A 5 Do not use for replacement birds None published mifur) over 14 weeks old

3-nitro-4- 5 Use as only source of organic Muscle,eggs,0.5;other byhydroxyphenylarsonic arsenic products^ acid(roxarsone)

Nitromide,sulfanitran 5 Do not use in egg-laying birds Nitromide,none

(Unistat-2) published;sulfanitran,0

Nitromide.sulfanitran 5 Do not use ih egg-laying birds, Nitromide,0.03;sulfanitran.0 (Unistat-3) use as only source of organic ;arsenic, same as above arsenic_^ ___

Novobiocin(Albamix) 4 Do not use in egg-laying birds 0 Nystatin _ 0 50-100g/900 kg feecf 0

Oleandomycin Do not use in egg-laying birds Edible tissues,0.15 Ormetoprim 5 Do not use in birds over 16 week Edible tissues,0.1 old

Oxytetracycline 0 50-200 g/900 kg feed Kidney,3; muscle,liver,fat,skin, 1

Penicillin JΛ- 25 g 900 kg feed Edible tissues, egges,0

Penicillin Do not use in egg-laying birds 0 powder(buffered) Robenidine~ Do not use in egg-laying birds Skin and fat,0.2;other edible hydrochloride tissues,0.1 (Robenz)

Spectinomycin Do not use in egg-laying birds Edible tissues,0.1

Streptomycin Administer not more than 4 days. Edible tissues, eggs,0 hydrochloride or do not use in egg-laying birds streptomycin sulfate

Sulfachloropyrazine,s 4 Do not use in birds producing 0 odium monαhydrate eggs for human food

Sιdfadimethoxine,orm5 Do not use in birds over 16 weeks 0.1 for both drugs etoprim(Rotenaid) old

Sulfanitran Do not use in egg-laying birds 0

Sulfaquinoxaline (SQ 10 Do not use in birds producing None published 40,Sulquin-40) eggs for human food

Sulfomyxin Do not administer to egg-laying 0 birds

Tetracycline_ 0.25 Tylosin phosphate 5* Feed at 0-5 days of age; 2^nd Edible tissues, eggs,0.2 administration is fed24-48 hrs at3-5 weeks of age

Reference in this connection may be made to John's Veterinary Pharmacology & Therapeutics, Ed. Nicholas, H. Booth, Leslie E.Macdonald, Kalyani publishing, Ludhiana, 5^th edition, 1984, 1083-84.

The European Union international conference on antimicrobial resistance that was held in ^' Copenhagen (Refer to International Pig Topics, Vol. 3, No.7, 1998, 3) came to several interesting conclusions. The Copenhagen document concluded that most of the delegates felt that the use of antimicrobials for growth promotion could not be justified and growth promoting antimicrobials should be replaced by safer non-antimicrobial alternatives including better farming practice.

Hence it is clear that a safer and effective alternative growth promoter is needed for domestic animals especially for poultry, cattle, birds etc.

As per J. Walter (1980), the characteristic of an ideal growth promoter for domestic animals should have the following essential features:

• Improves performance effectively and economically.

• Not cause cross-resistance to other antibiotics at actual use levels. »Not be involved with transferable drug resistance

•Not cause deleterious disturbances to the normal gut flora

•Not be absorbed from the gut into edible tissues.

•Not be mutagenic or carcinogenic

•Not give rise to environmental pollution • it must be readily biodegradable.

•Be non-toxic to the animal and its human handlers.

In this context, reference may also be made to International hatchery practice. Published by P.A.P. UK, 1998, Vol.l2, No.8, p.3

DEVELOPMENT OF THE INVENTION: After studying the requirements in connection with the development especially of poultry industry, it was learnt that efficacy of the various types of weight promoters hitherto available differ. Further their cost also varies, sometimes proving unaffordable for poultry farmer.

The present invention has been developed to meet the above said requirements Therefore the main objective of the present invention is to provide an improved herbal composition useful for hepatoprotection and promoting weight gain in domestic animals Another objective of the present invention is to provide an improved herbal composition useful for hepatoprotection and promoting weight gain in domestic animals which is very potent, non toxic and safe.

Yet another objective of the present invention is to provide an improved herbal composition useful for hepatoprotection and promoting weight gain in domestic animals which mixes well with feed and ensures homogeneous mixing Still another objective of the present invention is to provide an improved herbal composition useful for hepatoprotection and promoting weight gain in domestic animals which does not require any withdrawal period before slaughter and there are no toxic residues retained in the bird meat.

Yet another objective of the present invention is to provide an improved herbal composition useful for hepatoprotection and promoting weight gain in domestic animals which does not cause deleterious disturbances to the normal gut flora. Further another objective of the present invention is to provide an improved herbal composition useful for hepatoprotection and promoting weight gain in domestic animals which is not absorbed from the gut into edible tissues.

Still another objective of the present invention is to provide an improved herbal composition useful for hepatoprotection and promoting weight gain in domestic animals which is not mutagenic or carcinogenic

Yet another objective of the present invention is to provide an improved herbal composition useful for hepatoprotection and promoting weight gain in domestic animals which does not give rise to environmental pollution and is readily biodegradable.

Another objective of the present invention is to provide an improved herbal composition useful for hepatoprotection and promoting weight gain in domestic animals which is non-toxic to the animal and its human handlers.

In Ayurveda (Indian system of medicine) recommends several medicinal plants useful as herbal feed supplements which are also effective in curing ailments of the liver. Ayurvedic literature like "Vaidya Chintamani, Bhava Prakash Niganthu, Yoga Ratnakara and Charaka Samhita" have references to the herbs capable of being used in the treatment of liver disorders. The present invention has been developed based on our finding that a synergistic hepatoprotective and weight gain promoting composition can be formulated to meet the above mentioned objectives employing plants judiciously selected from the families of Acanthaceae, Euphorbiaceae, Solanaceae and Nyctaginaceae. From the above said families of the plants used in Ayurveda for hepatoprotective and weight gain promoting in domestic animals we have selected according to the present invention the following plants.

(l)ANDROGRAPHIS PANICULATA

(2)SOLANUM NIGRUM (3)PHYLLANTHUS AMARUS

(4) BOERHAA VIA DIFFUSA

The details of the information about the above said plants which are available in public domain are given below.

1) ANDROGRAPHIS PANICULA TA Part used: Whole plant

Botanical description: Erect or procumbent, herbs or under-shrubs, the branches are often tetragonous. Leaves entire. Flowers in axillary and terminal, often panicled, racemes, unilateral bracts small, bracteoles minute or absent. Calyx 5-partite, Corolla small, tubular at base. prominently 2-lipped, the upper slightly bifid, the lower usually deflexed, 3-Iobed and coloured. Stamens 2 filaments broad, anthers 2-celled. Ovary 2-celled, ovules 2-6 in each cell, style slender. stigma minutely bifid. Fruit an oblong or. elliptic capsule compressed at right angles to the septum,

4-12 seeded, the retinacula acute or rounded. Seeds hard, deeply rugosely pitted glabrous. (Ref.

Matthew K.M., "The Flora of Tamil Nadu Carnatic", Pub. The Rapinat Herbarium, Tiruchirapalli.

India. 1983, Vol. 2, 1152) Medicinal" uses: The plant is bitter, acrid, cooling, vulnerary, antipyretic, antiperiodic, anti- mflammatory, expectorant, depurative, sudoforic, anthelmintic, digestive and stomachic. It is useful in rryperdypsia, burning sensation, wounds, ulcers, chronic fever, malarial and intermittent fevers. irLflairirnations, cough, bronchitis, skin diseases, leprosy, pruritis, intestinal worms, flatulence colic. diarrhoea, dysentery, hemorrhoids (Wealth of Asia ,NISCOM, CSIR, New Delhi, 1996 -.Indian Medicinal Plants, A compendium of 500 species. Orient Longman, 1993; Vol. 1: 135; Nadakarni K.

M. Hie Indian Materia Medica, Pub. Bombay Popular Prakashan 1 93; Vol. 1 : 101)

Phytochemistry: The main constituents of Andrographis paniculata are diterepene lactones, viz.. andrographolide, neoandrographolide, andrographiside and flavones viz., oroxylin, wogonin, andrographidines A, B, C, D, E, F (Indian Herbal Pharmacopoeia, Vol. I, p.18-29. Joint Publication of IDMA (Mumbai) and RRL (Jammu-Tawi; Matsuda T, et al "Cell differentiation - Inducing Diterpenes from Andrographis pa iculata Nees", Chem. Pharm. Bull, 1994, 42(6),

1216- 1225)

Pharmacology: Hepatoprotective effect of andrographolide (the major active diterpenoid lactone of the plant Andrographis paniculata) was studied in many experimental models. In a study a dose of 300 mg/kg of the extract was selected to study hepatoprotective action against CC14-induced liver damage, was found to be effective in preventing liver damage, which was evident by morphological, biochemical and functional parameters (Rana, A.C; Avadhoot, Y;

"Hepatoprotective effects of Andrographis paniculata against carbon tetrachloride-induced liver damage", Archives of Pharmacological Research, 1991; 14(1), 93-95). Andrographolide was found to be more potent than silymarin, a known hepatoprotective drug (Visen, P.K.S; Shukla, B; Patnaik,

C.K; Dhawan, B.N; "Evaluation of hepatoprotective activity of andrographolide isolated from the plant Andrographis paniculata". Proceedings 24 , Indian Pharmacological Society, Conference,

1991; 12)

2) SOLANUM NIGRUM Part used: Whole plant

Botanical description: An erect annual herb,, -stem glabrous or pubscent, much divaricately branched. Leaves numerous, ovate, lanceolate, subacute or accuminate, glabarous, thin entire.

Flowers 3-8 in extra-axillary drooping sunbumbellate cymes; fruits purlish black or reddish berries.

Seeds many, discoid, minutely pitted yellow (Indian Medicinal Plants, A compendium of 500 species, Orient Longman, 1993,5,160-163)

Medicinal uses: The plant used as an antispasmodic and in the treatment of cirrhosis of the liver.

Berries are considered to posses tonic, diuretic and cathartic properties and are useful in heart diseases (Wealth of Asia ,NISCOM, CSIR. New Delhi, 1996 Vol. 1: 135)

Phytochemistry: The plant contains protein, fat, minerals, carbohydrates, riboflavin, nicotinic acid. vitamin-c, carotene, glucose, fructose, steroidal alkaloids viz. solamargine, solasonine, solanigrine. etc. (Wealth of Asia ,NISCOM. CSIR, New Delhi.1996)

Pharmacology: In a study conducted on rats there was increased feed consumption and increase in body weight in female rats which received Solanum nigrum cooked leaves in feed when compared to control group. Further, Solanum nigrum improved the quality and quantity of nutrients like Vit. C, β carotene, protein and aminoacids in feed which probably favour on increase in the activity of liver drug metabolizing enzymes. In another study there was increase in body weight, liver weight and liver protein in female weaning rats which received Solanum nigrum extract when compared to control group (Saxena A and NK Garg Ind J.exp. Bio., 1979 17(7),662-664).

The hepatoprotective activity of Solanum nigrum crude plant extracts may be due to their ability to suppress the oxidative degradation of DNA in the tissue debris. (Sultana S., et a! "Crude extracts of hepatoprotective plants, Solanum nigrum and Cichorium intybus inhibit free radical-mediated DNA damage.^" J. ofEthnopharmacology, 1995, 45, 189-192). 3) BOERHAA VIA DIFFUSA Part used: Whole plant Botanical description: A perennial diffuse herb with stout root stock and many procurement branches; leaves simple, opposite, short petioled in unequal pairs, ovate-oblong, acute or obtuse, subcordate at base, glabrous above and whitish beneath ; flowers pale rose colored, small, short stalked, in irregular clusters of terminal panicles at the ends of branches; fruits highly viscid, easily detachable, one-seeded, indehiscent with a thin peήcsrp(Indian Medicinal Plants, A compendium of 500 species, Orient Longman, 1993 ; Vol. 1 : 253-255)

Medicinal uses: The plant is bitter, astringent, cooling, anthelmintic, diuretic, emetic, expectorant. It is used in the treatment of jaundice (Wealth of Asia ,NISCOM, CSIR, New Delhi, 1996 lndian Medicinal Plants , A compendium of 500 species. Orient Longman, 1993; Vol. 1: 253-255) Pbytochemistry: The major constituent namely Punarnavoside is found in 0.03-0.05%.Other minor consdtuents include rotenoids viz., boeravinones A,B,C,D, & E ; lignans liridodendrin etc. Pharmacology: The aqueous extract of Boerhaavia diffusa roots (2 ml/kg rat administration) has more hepatoprotective activity than powder forrri. probably due to the better absorption of the liquid form through the intestinal tract (Rawat AKS et al "Hepatoprotective activity of Boerhaavia diffusa L. roots a popular Indian ethnomedicine^" J. ofEthnopharmacology, 1997,56,61 -66). 4) PHYLL4NTHUS AMARUS Part used: Whole plant

Botanical description: A small erect herb, stem terete, younger parts rough; leaf elliptic oblong to obvate; Flowers axillary, proximal 2-3 axils with unisexual 1-3 male flowers and all succeeding axils with bisexual cymules. Male flowers-pedicel long, calyx 5, sub equal, oblong, elliptic, apex acute, hyaline with unbranched mid rib; disc segments 5, rounded, stamens 3. filaments connate. Female flowers-pedicel long, calyx lobes 5, ovate-oblong, acute at apex; disc flat deeply 5 lobed, lobes often toothed at apex, styles 3, free, shallowly bifid at apex. Capsule oblate and rounded, seeds triangular with longirudinal ribs (Bagchi G.D, Srivastava G.N , Singh S.C, Distinguishing features of medicinal herbaceous species of Phyllanthus occuring in Lucknow District (U.P) India. Int. J. Plwrmacognosy, 1992. 30(3), 161-168)

Medicinal uses: The plant is bitter, astringent, cooling, diuretic, stomachic, febrifuge and antiseptic. It is useful in dropsy, jaundice, diarrhoea, dysentery, intermittent fevers, diseases of urino-genital system, scabies ulcers and wounds (Wealth of Asia, NISCOM, CSIR, New Delhi. 1996: Indian Medicinal Plants. A compendium of 500 species, Orient Longman. 1995; Vol. 4: 252- 255; Bhava prakash nigunia. Motilal banarsidas publications, 1998, 273) Phytochemistry: The major lignans Phyllanthin and Hypophyllanthin has been reported to exhibit antihepatotoxic activity. Lignans - a diarylbutane, Phyllanthin and an aryltetrahydronaphthalene, hypophyllanthin (Indian Herbal Pharmacopoeia, pub: Regional Research Laboratory, Jammu Tawi & Indian Drug Manufacturer's Association Mumbai, 1999, 2, 85 - 92) Pharmacology: Phyllanthus amarus at the dose of 0.66 g/kg in each rat orally was found to be effective, not only against liver damage from CC14 in rats but also effective in the liver regeneration (Sane R.T, Kuber V.V, Chalissery M.S., Menon S, Hepatoprotection by Phyllanthus amarus and Phyllanthus debili in Ccl4 induced liver dysfunction, Current Science, 1995, 68 (12), 1243 - 1246); In another study Phyllanthus amarus was tested on hepatitis B virus. The alcoholic extracts were effective against HBV antigens, the butanol extract being the most potent. The active fractions inhibited the interaction between HBsAg/HbeAg and corresponding antibodies suggesting anti- HBs, anti-Hbe-like activity and also an effect on HBV-DNA. ("In- vitro effect of Phyllanthus amarus on hepatitis B virus", Mehrotra R et al., Indian J. Med. Res. 1991, 93(A), 71-73]. The crude powders of the above mentioned plants were administered individually to domestic animals ( Poultry birds) to determine the hepatoprotective and weight gain promoting activities in the following manner

Day old chicks were obtained from M/s. Venkateshwara Hatcheries (Vencobb hybrid line) with equal sex ratio for the study and divided into control and treated groups. Control group received the feed (Lipton's Gold Mohur feed from Hindustan Lever Limited) and treated groups received the same feed mixed with the required doses of the test material( namely the above said plants ). The birds were fed for a period of 42 days. Feed consumption ratio, body weight, livability were recorded as the parameters. Additional parameters like Bursa weight, serum glutamate pyruvate transami ase (SGPT) and dressing percentage were also recorded. The results are shown in Table 3 ^~ TABLE-3

Group Test Strain of Feed used Avg. b.wt. (g) + Improvement Avg. FCR + Livability material bird SEM (%) SEM (%)

250 g/ton of feed

I Control 1665 + 1S5 0 2.128 + 0.029 92.46 π Plant 1 * 1764 + 160 5.9 2.113 ± 0.021 94.83 m Plant 2*

-hvbrid line Mohur 1800 ± 158 8.1 2.119 ± 0.031 91.64 rV Plant 3* feed 1810 + 149 8.7 2.012 + 0.026 95.32 V^"" Plant *- 1797 + 163 7.9 2.216 + 0.031 94.21 n = 50. In the above table Plant 1 relates to Andrographis paniculata;, Plant 2 relates to Solanum nigrum; Plant 3 relates to Phyllanthus amarus and Plant 4 relates to - Boerhaavia diffusa

From the information given in the Table 3 we have observed that above plants have not only hepatoprotective activity but also weight gain promoting properties in domestic animals such as poultry birds. Based on our above said observations we have developed the composition which is effective as hepatoprotective and weight gain promoter in domestic animals such as poultry birds.

Accordingly, the present invention provides, a herbal composition useful for hepatoprotection and promoting weight gain in domestic animals comprising

(i) 25- 35% w/w of the total weight of the composition of the powder of t e plant of Andrographis paniculata

(ii) 20-30% w/w of the total weight of the composition of the powder of the plant of Phyllanthus amarus (Hi) 20-30% w/w of the total weight of the composition of the powder of the plant of Solanum nigrum and

(iv) 15- 25% w/w of the total weight of the composition of the powder of the plant of Boerhaavia diffusa

In a preferred embodiment of the present invention any part of the above said plants may be used individually or all the parts together as per the requirements .

According to another feature of the invention the composition may be formed into any oral dosage forms like, tablets, bolus, capsules, syrups and liquids etc. For facilitating making the composition in the above said dosage forms, pharmaceutical excipients, which are usually employed to prepare such oral dosage form may be employed . By way of examples recipients like starch, pre- gelatinized starch, dicalcium phosphate or a mixture thereof may be used.

According^" to another feature of the present invention a herbal composition may also contain preservatives which may be selected from propyl paraben sodium, methyl paraben sodium or bronopol or a mixture thereof and the like .

According to yet another feature of the present invention there is provided a process for preparing the herbal composition useful for hepatoprotective and weight gain promotion in domestic animals which comprises

(i) mixing a first mixture of 25-35% w/w of the powder of the plant Andrographis paniculata and

20-30% w/w of the powder of the plant Phyllanthus amarus then, mixing the resultant first mixture with (ii) a second mixture of 20-30% w/w of the powder of the plant of Solanum nigrum and 15-25% w/w of the powder of the plant of Boerhaavia diffusa (iii) the powders having a mesh size of 5 to20 and

(iv)the resulting composition is finally pulverized by conventional methods to a mesh size of 70-

100 and the composition having a mesh size of 70- 100 In a preferred embodiment of the invention for making the powders any part of the above said plants individually or all the parts together may be used as per the requirements

Before making the powders , the parts of the plants or the whole plants are cleaned to avoid foreign materials , cut and chopped to make it easy to make the powders ..

According to another feature of the invention the composition may be formed into any oral dosage forms like, tablets, bolus, capsules, syrups and liquids etc by conventional means . For facilitating making the composition in the above said dosage forms, pharmaceutical excipients, which are usually employed to prepare such oral dosage form may be employed . By way of examples recipients like starch, pre-gelatinized starch, dicalcium phosphate or a mixture thereof may be used.

According to another feature of the present invention a herbal composition may also contain preservatives which may be selected from propyl paraben sodium, methyl paraben sodium or bronopol or a mixture thereof and the like .

The invention is described in detail in the Examples given below which are provided by way of illustration only and therefore should not be construed to limit the scope of the invention.

Example 1

An amount of 150gms (30%w/w) of Andrographis paniculata (whole plant) and lOOg s (20%/w) of Phyllanthus amarus (whole plant) were cleaned and chopped in a chopping machine to obtain a mesh size of 8 to 10 mesh and mixed together . Similarly 150 gms (30%w/w) of Solanum nigrum whole plant after cleaning and chopping was powdered in a hammer mill to a sieve size of 8 to 10 mesh was mixed with lOOgms (20% w/w) of the powder having the mesh size of 8 to 10 of the plant Boerhaavia diffusa, (whole plant). The above said two mixtures were mixed together and the resulting composition was pulverised in a pulveriser to a sieve size of 70 to 100 The above composition for convenience is henceforth referred to as Composition A

Each of the two above said compositions were subjected to the following trials.

Example 2 An amount of 125gms (25%w/w) of Andrographis paniculata (whole plant) and 150gms (30%/w) of Phyllanthus amarus (whole plant) were cleaned and chopped in a chopping machine to obtain a mesh size of 8 to 10 mesh and mixed together . Similarly 100 gms (20%w/w) of Solanum nigrum whole plant after cleaning and chopping was powdered in a hammer mill to a sieve size of 8 to 10 mesh was mixed with 125gms (25%w/w) of the powder having the mesh size of 8 to 10 of the plant Boerhaavia diffusa, (whole plant). The above said two mixtures were mixed together and the resulting composition was pulverised in a pulveriser to a sieve size of 70 to 100 5 The above composition for convenience is henceforth referred to as Composition B

Each of the two above said compositions were subjected to the following trials.

Experimentally aflatoxicosis was induced with aflatoxicin Bi, on broilers and efficacy of two Compostions (Example 1 and Example 2) as hepatoprotective and weight gain promoter was o assessed in 300 birds. The various parameters studies employed were weekly gain in body weights, feed consumption and FCR. Biochemical assessment was made using SGOT, SGPT, Albumin, Globulin and Albumin globulin ratio, Haematological parameters were also included like haemoglobin, packed cell volume. The humoral acquired immunity and cell mediated immunity were also included to evaluated the immune status. Gross and histopathological lesions of the 5 organs collected namely liver, kidney, bursa of Fabricius, etc. were also recorded. Composition B (Example 2) treated group showed significant improvement over the aflatoxin fed birds in terms of body weight gain, feed consumption, FCR, etc. The im une status of the birds revealed improved cell mediated and humoral immune response. These results can be seen in the Table 4 0 Table 4 Showing the efficacy of the Compositions A & B (Example 1 and Example 2 ) in aflatoxin induced hepatotoxicity

Test 1 shows- Example 1 (Composition A) Test 2 shows- Example 2 (Composition B)

The above study clearly indicates that both the Compositions are effective in protecting chicks against aflatoxin induced hepatotoxicity. However since Composition B (Example 2) was more potent it was selected for further studies. Composition B (Example 2)was tested for its efficacy at different dosages for its hepatoprotective and weight gain promoter activity in two different dosages to chicks (Vencobb hybrid line) Control group received the feed (Lipton's Gold Mohur feed from Hindustan Lever Limited) and treated groups received the same feed mixed with the required doses of the test material. The birds were fed for a period of 42 days. Feed consumption ratio, body weight, livability were recorded as the parameter. Additional parameters like Bursa weight, serum glutamate pyruvate transaminase (SGPT) and dressing percentage were also recorded. Composition B (Example 2) at 250g/ton dosage afforded maximum hepatoprotection and weight gain which was comparable with a dosage of 500g/tonne of feed. The results are given in Table 5

Table 5

n = 60

Test 2 shows the Example 2 ( Composition B ) - 250 g/ton ;

Test 2- shows the Example 2 ( Composition B ) 500g/ton

Form the information provided in Table 3 ( for individual plants - column 6 ) and Table 5 ( for the mixture of the plants - Column 6) it would be observed that the improvement percentage has increased 3 to 4 times when the mixture of the plants as per the present invention is administered . This improvement is due to the synergistic combination of the plants used in the specified quantities and method of preparing the composition .

Accordingly the composition of the present invention is not a mere admixture of the ingredients employed but is a result of a unexpected synergistic combination of the ingredients employed Study of acute toxicity of the Example 2(Composition B)

Sixteen Albino Swiss mice were taken, divided into two groups and fasted for 16 hours. One group served as the control and received the 'vehicle'. The second group received the feed supplement (Composition B ) powder as a 0.5% suspension in crystalline microcellulose at a dose of 3000mg/kg mice body weight. There were no symptoms of toxicity (gross observation) and death still 72 hours. 56mg/kg mice body weight was found to be the effective antihepatotoxic dose and the therapeutic index was found to be >53 times. Hence it can be inferred that the feed supplement (Composition B) is very safe.

Comparative evaluation of the powder and liquid forms of the Composition B :

The composition B was tested for its efficacy as hepatoprotective in its crude drug form and as a liquid The liquid form of the Composition B was prepared by mixing the powder with water

Male albino rats of either sex weighing 200-250gms were taken and the composition B, both the powderf 250gm / ton of feed )and the liquid forms( (@ 2.5 mg / kg of body weight) separately were administered for two weeks, mixed with feed or in drinking water. Carbon terra chloride was administered after 48 hours to induce hepatotoxicity or liver damage. The data was analysed using ANOYA and Student's t-test. And the result is given below

Group Control CTC Composition B Composition B (as powder) (as liquid)

SGPT(U L) 55+8 1278+140* 160+78⁺ 348+83"

* pO.OQOl compared to other groups -p<0.005

Field trials of Composition B in powder form were conducted at various agroclimatic condition, feeds and strains of birds in India on layers and broilers have shown an improvement in body weight, livability and production number. Composition B has also shown to reduce the egg shell breakage and reduce mortality in layers as can be seen from the Table 6 .

Table 6 RESULTS OF CONTROLLED/FIELD TRIALS IN BROILERS

* The flock was sold little earlier because of better market prices.

Z -Composition B C-CONTROL group * % improvement over control group

Conclusion: The results indicate that the performance enhancing activity of the composition B is very consistent and it is independent irrespective of strain of broiler, feed (branded/unbranded) and agro climatic conditions. The composition of the present invention also improves feed metabolism and digestion and thus helps in better growth, weight gain and general health. It acts in the following ways:

1. Increases secretion and flow of bile from the liver and thereby assists fat digestion and assimilation.

2. Shows potent hepatoprotective and anti-hepatotoxic activity, guards the liver parenchyma against toxic effects of toxins, antibiotics, -mthelrnintics etc.

3. Exhibits strong hepatogenic activity and helps in the regeneration of damaged liver tissue.

4.1n cases of liver damage indicating delayed BSP clearance, rise in bilirubin, low A/G ratio and prolonged prothrombin time (clotting time), therapy enables all the above parameters, to return to normal range.

5. Stimulates glycogenesis and glyconeogenesis in the liver. Also helps in restoring cholesterol metabolism in the liver. ό.Research has indicated that its supplementation in feed is effective not only in inhibiting damages caused by Aflatoxins in livestock but also in reversing the damaging effects of Aflatoxin with improved growth, weight gain, early maturity and better farm productivity.

Indications: Jaundice,Hepatitis.Ailatoxicosis,To improve the body resistance,Debility and general weakness,Toxaemia,During Convalescence,Fatty liver diseases,Leg weakness,For improving feed efficiency, general health and productiorLFor better livability,As a adjunct or after antibiotic/anthelmintic/chemical drugs therapy to restore normal detoxification and liver functions.

Advantages of the invention

Special benefits of the invention:

Broilers Layers

1. 6-10% higher weight gain Attainment of uniform sexual maturity

2. 4-9% improvement in FCR Better hen housed production

3. Nullifies the ill-effects of feed related toxins, i.e.Better FCR Aflatoxins, chemicals and pesticides

4. Enhancement of general health and henceReduction in thin shell and egg breakages improvement in livability in the older birds

5. Shortens the growing period Nullifies the ill-effects of feed related toxins, i.e. Aflatoxins, chemicals and pesticides

Additionally, the present invention has other advantages such as

1.It is safe and non-toxic

2. Mixes well with feed-ensures homogeneous mixing

3. Highly cost effective 4.There is no withdrawal period required before slaughter and there are no toxic residues retained in the bird meat.

5. It is very potent, non toxic and safe

6. It does not cause deleterious disturbances to the normal gut flora which is not absorbed from the gut into edible tissues. 7. It is not to be mutagenic or carcinogenic

8. It does not give rise to environmental pollution and is readily biodegradable.

9. It is non-toxic to the animal and its human handlers.

Claims

We claim:-

1. A herbal composition useful for hepatoprotective and promoting weight gain in domestic animals comprising (i) 25- 35% w/w of the total weight of the composition of the powder of the plant of Andrographis paniculata

(ii) 20-30%) w/w of the total weight of the composition of the powder of the plant of Phyllanthus amarus

(in) 20-30% w/w of the total weight of the composition of the powder of the plant of Solanum nigrum and

(iv) 15- 25% w/w of the total weight of the composition of the powder of the plant of Boerhaavia diffusa and the composition having a mesh size of 70- 100

2. A herbal composition as claimed in claim 1 wherein any part of the above said plants is used individually or all the all the parts together as per the requirements

3.A herbal composition as claimed in claims 1 & 2 wherein the composition is formed into any oral dosage forms like, tablets, bolus, capsules, syrups and liquids etc.

4 A herbal composition as claimed in claims 1 to 3, wherein the composition contains pharmaceutical excipients, which are usually employed to prepare such as starch, pre-gelatinized starch, dicalcium phosphate or a mixture thereof .

5. A herbal composition as claimed in claim 3 & 4 wherein the composition also contains preservatives selected from propyl paraben sodium, methyl paraben sodium or bronopol or a mixture thereof and the like.

6. A process for the preparation of a herbal composition as claimed in claims 1 to 5 which comprises

(i) mixing a first mixture of 25-35% w/w of the powder of the plant Andrographis paniculata and 20-30% w/w of the powder of the plant Phyllanthus amarus then, mixing the resultant first mixture with

(ii) a second mixture of 20-30% w/w of the powder of the plant of Solanum nigrum and 15-25% w/w of the powder of the plant of Boerhaavia diffusa (iii) the powders having a mesh size of 5 to20 and

(iv)tbe resulting composition is finally pulverized by conventional methods to a mesh size of 70- 100