A polyherbal Anti-cancer formulation

Abstract

The present invention is all about a polyherbal anti-cancer formulation. The present invention relates to a polyherbal anti-cancer formulation comprising various part of plants. The present invention provides a polyherbal anti-cancer formulation comprising Zingibar officinale, Tinospora cordifolia, Swertia chirayita, Picrorhiza kurroa, Cissampelos pareiras, Cyperus rotundus, Cedrus deodara, Marsdenia tenacissima, Holarrhena antidysenterica and Hemidesmus indicus as herbal ingredients. The present invention also relates to a polyherbal anti-cancer formulation for treatment of breast cancer and process for preparing the same.

Description

[0001] A POLYHERB AL ANTI-CANCER FORMULATION

[0002] FIELD OF THE INVENTION

[0003] The present invention is all about a polyherbal anti-cancer formulation. More particularly, the present invention relates to a polyherbal anti-cancer formulation comprising various part of plants. The present invention provides a polyherbal anticancer formulation comprising Zingibar officinale, Tinospora cordifolia, Swertia chirayita, Picrorhiza kurroa, Cissampelos pareiras, Cyperus rotund s, Cedrus deodara, Marsdenia tenacissima, Holarrhena antidysenterica and Hemidesmus indicus as herbal ingredients. The present invention also relates to a polyherbal anticancer formulation for treatment of cancer and process for preparing the same.

[0004] BACKGROUND OF THE INVENTION

[0005] Cancer is a term used to describe a group of diseases characterized by abnormal cell proliferation that has the potential to infiltrate or spread to other parts of the body. A lump, unusual bleeding, a persistent cough, unexplained weight loss, and a change in bowel movements are all possible indications and symptoms. In 2015, around 90.5 million people were diagnosed with cancer. As of 2019, over 18 million new cases are reported each year. The deaths of approximately 8.8 million people each year (15.7 percent of deaths). Around 1, 65, 000 children under the age of 15 were diagnosed with cancer in 2012. Cancer risk rises dramatically as people get older, and several malignancies are more common in developed countries.

[0006] Lung cancer, prostate cancer, colorectal cancer, and stomach cancer are the most frequent cancers in men. Breast cancer, colorectal cancer, lung cancer, and cervical cancer are the most frequent cancers in women. Except for skin cancers, breast cancer is the most frequent cancer in women around the world. Each year, it accounts for roughly 30% (or 1 in 3) of all new female malignancies.

[0007] Breast cancer is a disorder in which the cells of the breast get uncontrollably large. There are various types of breast cancer. The type of breast cancer is determined by which cells in the breast become cancerous. Breast cancer can start in a variety of places in the breast. Lobules, ducts, and connective tissue are the three primary components of a breast. The glands that generate milk are known as lobules. The ducts are tubes that transport milk from the breast to the nipple. Everything is held together by connective tissue, which is made up of fibrous and fatty tissue. Breast cancer usually starts in the ducts or lobules. Breast cancer can spread to other parts of the body via blood and lymph vessels. Breast cancer is said to have metastasized when it spreads to other regions of the body.

[0008] Chemotherapy, radiation therapy, surgery, and immunotherapy are common therapies for breast cancer, but they have a number of drawbacks, including systemic side effects, a lack of site specificity, hair loss, gastro-esophageal reflux syndrome, and poor solubility and absorption. Most of the allopathic anti-cancer drugs can not differentiate between cancerous cell and normal cell. This leads to significance side effects like lower blood count, hair loss, mouth ulcers, nausea and diarrhea. As a result of these side effects, many patients are turning to complementary and alternative medicines (CAMs), including herbal therapies.

[0009] Ayurveda, the oldest Indian indigenous medicine system of plant drugs is known from very early times for preventing or treatment of various tumors using the natural drugs. Herbal treatments are more effective and safe, have a less side effects. Medicinal plants have different types of phytoconstituents which may exert their effectiveness in cancer treatment. Polyherbalism gives various benefits not accessible in single herbal formulations due to synergism. Therefore, need to develop liquid solution that contain mixture of 10 herbs and cow urine have some phytochemicals from each plant and are imbed with goodness of miraculous properties of cow urine. Phyto-chemicals from plants like tannins, flavonoids, triterpenoids, and alkaloids have been shown to possess anticancer activity.

[0010] Bioavailability is the ability of a drug or other substance to be absorbed and used by the body. The rate of the drug absorbed determines the onset of action of the drug. The various formulations of the drugs take different time for getting absorbed and showing their effect on the site of action. Tablet and capsule formulation is the most used formulation, But the tablets and capsule takes time for absorption as first they need to be dissolve and disintegrate before absorption. To overcome the disadvantages of tablets and capsules, oral liquid formulations of different drugs have been developed. Liquid formulations of drugs have faster absorption rate and ultimately have a higher bioavailability. Also, liquid formulations are in better compliance with children and patients of various age groups as they are easy to administer.

[0011] Therefore, inventors of the present invention has surprisingly developed a polyherbal liquid formulation from various part of plants for breast cancer treatment which is better alternative with no side effects to the allopathic and chemotherapeutic treatment of cancer.

[0012] OBJECTIVES OF THE INVENTION

[0013] The main objective of the present invention is to provide a polyherbal anti-cancer formulation.

[0014] Another objective of the present invention is to provide a polyherbal anti-cancer formulation which is safe and have less side effects.

[0015] Yet another objective of the present invention is to provide a polyherbal anticancer formulation which is having better patient compliance.

[0016] Another objective of the present invention is to provide a polyherbal anti-cancer formulation which provides anti-cancer activity.

[0017] SUMMARY OF THE INVENTION

[0018] The main aspect of the present invention is to provide a polyherbal anti-cancer formulation. Another aspect the present invention is to provide polyherbal anti-cancer formulation comprising herbal ingredients Zingibar officinale, Tinospora cordifolia, Swertia chirayita, Picrorhiza kurroa, Cissampelos pareiras, Cyperus rotund s, Cedrus deodara, Holarrhena antidysenterica, Marsdenia tenacissima, Hemidesmus indicus and cow urine.

[0019] Another aspect the present invention is to provide polyherbal anti-cancer formulation and process of preparation thereof.

[0020] BRIEF DESCRIPTION OF DRAWINGS

[0021] Fig i : illustrate phytochemical test of polyherbal anti-cancer formulation.

[0022] Fig 2: illustrates LC-MS analysis of polyherbal anti-cancer formulation.

[0023] Fig 3 : illustrate cell line of polyherbal anti-cancer formulation.

[0024] Fig 4: illustrate weight variation data during treatment phase of anti-cancer study.

[0025] Fig 5 : illustrate Gross necropsy data of mammary gland tumor

[0026] Fig 6: illustrate tumor volume data

[0027] Fig 7: illustrate Mda level from mammary / tumor homo generate

[0028] Fig 8 : illustrate SOD level from mammary / tumor homo generate.

[0029] Fig 9 : illustrate graphical representation of estimation of serum TNF-a level.

[0030] Fig 10: illustrate graphical representation of Liver function test via biomarkers Fig 11: illustrate graphical representation of Kidney function test via biomarkers

[0031] DETAILED DESCRIPTION OF THE INVENTION

[0032] The detailed description set forth below is intended as a description of exemplary embodiments and is not intended to represent the only forms in which the exemplary embodiments may be constructed and / or utilized. The description sets forth the functions and the sequence of steps for constructing and / or operating the exemplary embodiments. However, it is to be understood that the same or equivalent functions and sequences which may be accomplished by different exemplary methods are also intended to be encompassed within the spirit and scope of the invention. As defined herein, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs.

[0033] Although any process and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described.

[0034] As stated in the present invention herein, the singular forms “a,” “an” and “the” specifically also encompass the plural forms of the terms to which they refer, unless the content clearly dictates otherwise. The term “about” is used herein to means approximately, in the region of, roughly, or around.

[0035] As stated herein, that it follows in a transitional phrase or in the body of a claim, the terms “comprise(s)” and “comprising” are to be interpreted as having an open ended meaning. That is, the terms are to be interpreted synonymously with the phrases “having at least” or “including at least”. When used in the context of a process, the term “comprising” means that the process includes at least the recited steps, but may include additional steps. When used in the context of a composition, the term “comprising” means that the composition includes at least the recited features or components, but may also include additional features or components.

[0036] As used herein term “formulation” or “composition” or “dosage” conveys the same meaning and can be used interchangeably.

[0037] As per one embodiment as defined herein, “polyherbal” means the use of more than one herb in a medicinal preparation.

[0038] The main embodiment of the present invention is to provide a polyherbal anti-cancer formulation. Another main embodiment of the present invention is to provide a polyherbal anticancer formulation comprising Zingibar officinale, Tinospora cordifolia, Swertia chirayita, Picrorhiza kurroa, Cissampelos pareiras, Cyperus rotund s, Cedrus deodara, Marsdenia tenacissima, Holarrhena antidysenterica, Hemidesmus indicus and cow urine.

[0039] Zingibar officinale

[0040] The present invention contains Zingiber officinale, belonging to family Zingiberaceae commonly known as “ginger” or “Sunth” is a spice consumed worldwide for culinary and medicinal purposes. Sunth consists of dried rhizome of Zingiber officinale Roxb. Widely cultivated in India, rhizomes dug in January- February, buds and roots removed, soaked overnight in water, decorticated, and sometimes treated with lime and dried. The plant has a number of chemicals responsible for its medicinal properties, such as ant arthritis, anti-inflammatory, antidiabetic, and appetite stimulant, anti-emetic and in motion sickness, antibacterial, antifungal, anticancer, etc.

[0041] As per another embodiment of the present invention, Rhizome part of Zingibar officinale is used for polyherbal anti-cancer formulation.

[0042] As per another embodiment of the present invention, Zingibar officinale is used preferably in the range of 1 to 50 mg, more preferably 1 to 30 mg and most preferably 1 to 20 mg.

[0043] Tinospora cordifolia

[0044] The present invention contains Tinospora cordifolia, belonging to family Menispermaceae commonly known as “Guduchi” or “Giloy” is a consumed worldwide for medicinal purposes. It is basically native to the Indian subcontinent and South East Asia. The plant has a number of chemicals responsible for its medicinal properties such as Alkaloids, Terpenoids, Lignans, Steroids and also has medicinal property like antioxidant activity, antimicrobial activity, antibacterial activity, antifungal activity, anti-diabetic activity, antistress activity.

[0045] As per another embodiment of the present invention, stem part of Tinospora cordifolia is used for polyherbal anti-cancer formulation.

[0046] As per another embodiment of the present invention, Tinospora cordifolia is used preferably in the range of 1 to 50 mg, more preferably 1 to 30 mg and most preferably 1 to 20 mg.

[0047] Swertia chirayita

[0048] The present invention contains Swertia chirayita, belonging to family Gentianaceae commonly known as “kirat tikta” is consumed worldwide for medicinal purposes. Swertia chirayita is cultivated from Bhutan to Kashmir and in the Khasi hills at 1200-1500m and it also can be grown in sub-temperate territories between 1500- 2100m altitudes. The plant used for treating fever, upset stomach, constipation, anorexia (appetite loss), skin diseases, intestinal worms and cancer.

[0049] As per another embodiment of the present invention, whole part of Swertia chirayita is used for polyherbal anti-cancer formulation.

[0050] As per another embodiment of the present invention, Swertia chirayita is used preferably in the range of 1 to 50 mg, more preferably 1 to 30 mg and most preferably 1 to 20 mg.

[0051] Picrorhiza kurroa

[0052] The present invention contains Picrorhiza kurroa, belonging to family Plantains commonly known as “Katuki” is a consumed worldwide for medicinal purposes. Picrorhiza is a plant that grows in the Himalayan Mountains. Picrorhiza is used for allergy, liver and upper respiratory tract, reduce fevers, and to treat dyspepsia, chronic diarrhea, and scorpion sting. Picrorhiza contains chemicals that might stimulate the immune system, kill cancer cells, and relieve inflammation (swelling).

[0053] As per another embodiment of the present invention, Rhizome part of Picrorhiza karroo is used for polyherbal anti-cancer formulation.

[0054] As per another embodiment of the present invention, Picrorhiza kurroa is used preferably in the range of 1 to 50 mg, more preferably 1 to 30 mg and most preferably 1 to 20 mg.

[0055] Cissampelos pareiras

[0056] The present invention contains Cissampelos pareiras, belonging to family Menispermaceae is usually known as “Ambastha” or “Laghu Patha” consumed worldwide for medicinal purposes. It grows primarily in the wet tropical biome. The plant Cissampelos pareiras contain chemicals like isoquinoline alkaloids along with few flavonoids, flavonoid glycosides, and fatty acids. The crude extracts of C. pareira have shown various pharmacological activities such as antipyretic, antiinflammatory, antiarthritic, antiulcer, antidiabetic, anticancer, antifertility, antimicrobial, antioxidant, antivenom, antimalarial, and immunomodulatory, etc.

[0057] As per another embodiment of the present invention, Root part of Cissampelos pareiras is used for polyherbal anti-cancer formulation.

[0058] As per another embodiment of the present invention, Cissampelos pareiras is used preferably in the range of 1 to 50 mg, more preferably 1 to 30 mg and most preferably 1 to 20 mg.

[0059] Cyperus rotundus

[0060] The present invention contains Cyperus rotundus, belonging to family Sedges commonly known as “Musta” is a consumed worldwide for medicinal purposes. Cyperus rotundus is a perennial plant that may reach a height of up to 140 cm. It is native to tropical and subtropical Old World, with a significant presence in Africa, southern and central Europe, and southern Asia, including India. Several chemical have been identified like cadalene, cyprotene, flavonoids, sesquiterpenes, terpenoids, mustakone, isocyperol, acyperone, rotundene, valecine.

[0061] As per another embodiment of the present invention, Rhizome part of Cyperus rotundus is used for polyherbal anti-cancer formulation.

[0062] As per another embodiment of the present invention, Cyperus rotundus is used preferably in the range of 1 to 50 mg, more preferably 1 to 30 mg and most preferably 1 to 20 mg.

[0063] Cedrus deodara

[0064] The present invention contains Cedrus deodara, belonging to family Pine commonly known as “Devdaru” is a consumed worldwide for medicinal purposes. Cedrus deodara widely used to treat inflammation, pain, spasm, diabetes, Herpes simplex virus type-1 and cancer. It is native to the Himalayan region, specifically in the western parts of the Himalayas. The Cedrus deodara wood contains cedeodarin, ampelopsin, cedrin, cedrinoside, and deodarin (3, 4, 5, 6-tetrahydroxy-8-methyl dihydroflavonol).

[0065] As per another embodiment of the present invention, Heart wood part of Cedrus deodara is used for polyherbal anti-cancer formulation.

[0066] As per another embodiment of the present invention, Cedrus deodara is used preferably in the range of 1 to 50 mg, more preferably 1 to 30 mg and most preferably 1 to 20 mg.

[0067] Marsdenia tenacissima

[0068] The present invention contains Marsdenia tenacissima, belonging to family Apocynaceae commonly known as “Murva” is a consumed worldwide for medicinal purposes. Tropical and subtropical dry and moist deciduous forests having annual rainfall between 1000 mm and 1500 mm are the natural habitat of Marsdenia. Approximately 196 chemical ingredients covering steroids, triterpenes and organic acids have been identified from different parts of Marsdenia tenacissima plant.

[0069] Marsdenia tenacissima used for the treatment of asthma, trachitis, tonsillitis, pharyngitis, cystitis, cancer, hypertention, pneumonia and drug or food poisoning. As per another embodiment of the present invention, Root part of Marsdenia tenacissima is used for polyherbal anti-cancer formulation.

[0070] As per another embodiment of the present invention, Marsdenia tenacissima is used preferably in the range of 1 to 50 mg, more preferably 1 to 30 mg and most preferably 1 to 20 mg.

[0071] Holarrhena antidysenterica

[0072] The present invention contains Holarrhena antidysenterica, belonging to family Apocynaceae commonly known as “Vatsakphal” or “kutaj” is a consumed worldwide for medicinal purposes. Holarrhena antidysenterica is widely used in traditional medical system for treatment of constipation, colic, and diarrhea and it is also potent to fight against cancerous cells. It is widely found in Indian subcontinent and the tropical Himalayas.

[0073] As per another embodiment of the present invention, seed of Holarrhena antidysenterica is used for polyherbal anti-cancer formulation.

[0074] As per another embodiment of the present invention, Holarrhena antidysenterica is used preferably in the range of 1 to 50 mg, more preferably 1 to 30 mg and most preferably 1 to 20 mg.

[0075] Hemidesmus indicus The present invention contains Hemidesmus indicus, belonging to family Apocynaceae commonly known as “Sariva” is a consumed worldwide for medicinal purposes. Hemidesmus indicus used in treatment of leprosy, skin diseases, fever, asthma, bronchitis, syphilis, pruritus and other urinary diseases, cancer, chronic rheumatism, and leucorrhoea. It is native to the tropical and subtropical regions of India. The roots of Hemidesmus indicus contain hexatriacontane, lupeol, its octacosanoate, a-amyrin, P-amyrin, its acetate and sitosterol.

[0076] As per another embodiment of the present invention, Root of Hemidesmus indicus is used for polyherbal anti-cancer formulation.

[0077] As per another embodiment of the present invention, Hemidesmus indicus is used preferably in the range of 1 to 50 mg, more preferably 1 to 30 mg and most preferably 1 to 20 mg.

[0078] All herbal ingredient were purchased from shree kamdhenu divya Aushadhi mahila sahkari mandali Jamnagar, Gujarat, India.

[0079] As per one embodiment of the present invention, cow urine is used in the polyherbal liquid anti-cancer formulation.

[0080] As per one embodiment of the present invention, cow urine is used for maintaining the pH in the present invention.

[0081] As per one embodiment of the present invention, polyherbal anti-cancer formulation can be prepared from infusion, decoction, maceration, percolation, oil infusion and distillation process.

[0082] As per the main embodiment of the present invention, polyherbal anti-cancer formulation can be prepared from distillation process. As per one embodiment of present invention, process of preparation of polyherbal anti-cancer formulation comprises the steps of: a) Weighing all herbal ingredients properly; b) Soaking all herbal ingredient in one liter cow urine for 8 hours; c) Adding equal quantity of cow urine into step (b) solution; d) Performing distillation of step (c) until all the liquid phase get distillate; e) Filtering the solution of step (d) to remove any foreign particle; f) Storing the solution of step (e) for further use.

[0083] As per one embodiment of the present invention, polyherbal anti-cancer formulation is safe and have less side effects.

[0084] As per one embodiment of the present invention, polyherbal anti-cancer formulation is effective in treatment of breast cancer.

[0085] The invention is further illustrated by the following examples which are provided to be exemplary of the invention and do not limit the scope of the invention. While the present invention has been described in terms of its specific embodiments, certain modifications and equivalents will be apparent to those skilled in the art and are intended to be included within the scope of the present invention.

[0086] EXAMPLES

[0087] EXAMPLE 1: OPTIMIZED BATCH FOR POLYHERBAL FORMULATION

[0088] Table 1: Optimized batch for liquid formulation

[0089] Process of Preparation a) All herbal ingredients were weighed properly; b) All herbal ingredient were soaked in one liter cow urine for 8 hours; c) The equal quantity of cow urine was added into step (b) solution; d) The distillation of step (c) was performed until all the liquid phase get distillated; e) The solution of step (d) was filtered to remove foreign particle; f) The solution of step (e) was stored for further used.

[0090] EXAMPLE 2: PHYTOCHEMICAL TEST OF POLYHERBAL ANTICANCER FORMULATION

[0091] Polyherbal anti-cancer formulation undergoes the phytochemical test and based on the test it was observed that tannins, flavonoids, triterpenoids, glycoside and alkaloids are present in herbal ingredients which leads to the polyherbal liquid formulation decrease the risk of breast cancer.

[0092] The phytochemical test of polyherbal anti-cancer formulation was seen in figure 1

[0093] EXAMPLE 3: LC / MS OF POLYHERBAL ANTI-CANCER FORMULATION

[0094] LC / MS analysis of polyherbal anti-cancer formulation was performed to separate, quantify and identify the compounds present in the herbal plants and based on the LC / MS analysis of polyherbal liquid formulation it discloses the presence of Zingiberene, Shagol, Tinosporide, Swerchirin, Amarogentin, Swertiamarin, Cyprotundone, Alpha-atlantone, Gamma-atlantone, Pregnane, Holadysentrrine, Holadienone, Kurchessine compounds present in the plants used in polyherbal anticancer formulation.

[0095] EXAMPLE 4: IN VITRO ASSAY OF ANTI-CANCER FORMULATION BY

[0096] USING MCF-7 CANCER CELL LINE a) The cell culture was added in 28 well plate(l x 105cell / wall); b) The step (a) was incubated for 24 hour for attachment at 37 °C and 5% CO2; c) The step (b) was treated with polyherbal liquid formulation; d) The step (c) was incubated for 24 hours at 37 °C and 5% CO2; e) The 100 pl of MTT solution[(3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide)] was added into the step (d) wells; f) The wells of step (e) was incubated for 1 hour; g) The excess medium was removed from step (f) and two wash of saline water were used; h) The lying buffer (200 pl) was added into each well; i) The color was measured at wavelength 562 nm.

[0097] The minimum inhibitory concentration obtained during in-vitro assay of polyherbal anti-cancer formulation by using MCF-7 cancer was 188.031 ug / ml. The cell line formed by polyherbal anti-cancer formulation was seen in figure 3.

[0098] EXAMPLE 5: IN VIVO ASSAY OF POLYHERBAL ANTI-CANCER

[0099] FORMULATION

[0100] 1. Acute oral toxicity study

[0101] 2. Anti-cancer study

[0102] Table 2: Animal requirement for acute toxicity and cancer study EXAMPLE 6: EVALUATION PARAMETER FOR ACUTE ORAL

[0103] TOXICITY STUDY

[0104] In acute oral toxicity study, the groups of rats (normal and treatment) were taken. Treatments were administered orally in the rats and the study performed focused only on evaluating toxicity by observing any changes in the different parameters

[0105] RESULT

[0106] The parameters in acute oral toxicity performed in the above table 3 concludes that there were no death signs or toxicity developed in rats and neither were any significant difference in body weight and no pathological abnormalities found in normal and treated groups.

[0107] EXAMPLE 7: ANTI-CANCER STUDY OF ANTI-CANCER FORMULATION

[0108] DMBA (7,12- dimethyl benz [a] anthracene) was used for induction breast cancer in healthy female Sprague-Dawley rats weighing around 75-90gm and were six weeks old through the subcutaneous route.

[0109] 1. After 90- 110 days the tumor was formed and rat’ s body weight measurement was performed every week after DMBA induction.

[0110] 2. The tumor bearing rats were treated with herbal formulations for 30 days.

[0111] 3. Anti-cancerous drug were administered orally for 30 days until the day of sacrifice.

[0112] Table 4: Dose Distribution in groups

[0113] EXAMPLE 8: EVALUATION PARAMETER OF ANTI-CANCER

[0114] ACTIVITY 8.1. CHANGE IN BODY WEIGHT

[0115] 1. Weight variation data during treatment phase

[0116] The weight variation was calculated from day 91th because the tumor injected was formed after 90 days.

[0117] Table 5: Weight variation data during treatment phase Result

[0118] As per the results on day 121 G-3 Standard doxorubicin G-3 shows significant reduction in weight as compared to G-2 Disease control group, this happens due to cytotoxic effect of Doxorubicin drug. Meanwhile treatment group G-4, 5 & 6 shows no observable effect on weight variation during 30-days treatment period. Since chemotherapy often decrease the weight because it suppresses all rapidly dividing cells. But in our case we had observed that anticancer liquid formulation was not significantly effective in weight gaining process, but parallel this formulation has stable the weight for 30 days treatment.

[0119] 8.2 GROSS NECROPSY OF MAMMARY GLAND TUMOR

[0120] Table 10: Gross necropsy of mammary gland tumor

[0121] The gross necropsy of mammary gland tumor observed in the table 10 and figure 9 it was observed that mammary gland tumor was reduced during treatment phase control group in group 6 with dose of (4ml / day / kg) it was reduced to 10.18nm.

[0122] 8.3 TUMOR VOLUME DATA

[0123] Table 7: Tumor volume data

[0124] Based on the table 13 and figure 10 it was observed that the tumor volume was reduced during treatment phase control group in group 6 with dose of (4 ml / day / kg) it was reduced to 2.3 ± 0.42.

[0125] 8.4. MDA LEVEL FROM MAMMARY / TUMOUR HOMO GENATE

[0126] Table 8: Mda level from mammary / tumor homogenate

[0127] The reduction of MDA level was observed in table 14 and figure 11 and it was observed that MDA Level was reduced during treatment phase control group and in group 5 with dose of (3 ml / day / kg) it was highly reduced to 3.18 ± 0.482.

[0128] 8.5. SOD LEVEL FROM MAMMARY / TUMOUR HOMO GENATE

[0129] Table 9: SOD level from mammary / tumor homogenate

[0130] Based on the results, it was observed that there were increase in the SOD level in the treatment groups comparatively disease and standard group of polyherbal anticancer formulation.

[0131] 8.6. GROSS NECROPSY OF ORGANS

[0132] Gross necropsy of various organs like brain, liver, heart, lungs, kidney and ovary was observed and there was no damage part observed in the organs of disease control group and there was no damage seen during treatment phase control group.

[0133] 8.7. EVALUATION OF HISPATHOLOGICAL MAMARIAN TISSUE

[0134] It was observed that there were reduction in the tumor in the rats of treatment group and there was no formation of dysplasia was observed in the treatment groups.

[0135] 8.8. EVALUATION OF LIVER FUNCTION TEST VIA ESTIMATING

[0136] LIVER BIOMARKER SGOT & SGPT

[0137] Table 10: Liver function test via biomarkers

[0138] The estimation of liver function test was done via biomarkers like the SGOT and SGPT. The level of SGOT and SGPT level was reduced in treatment group with the Group 5- Treatment (3ml / day / kg) with 44.56 ± 9.71 of SGOT and 73.78 ± 12.11 SGPT (U / L) 8.9. EVALUATION OF KIDNEY FUNCTION TEST VIA ESTIMATING

[0139] KIDNEY BIOMARKER UREA, URIC ACID AND CREATININE

[0140] Table 11: Kidney function test via biomarkers

[0141] The evaluation of liver function test was done via using biomarkers like the urea, uric acid and creatinine and it was observed that in serum urea during our treatment group there was significant decrease in serum urea as compare to the disease control group in dose dependent manner. In the uric acid, there was significant increase observed in higher dose of treatment control group as compare to disease control group and there was no significant difference in creatinine level in DMBA disease control group as compare to the normal control group as well as treatment control groups. During all these observation it was concluded that anticancer liquid formulation serves as kidney organ protector in treatment control group.

Claims

ClaimsWe claim,1. A polyherbal anti-cancer formulation comprising of : a. A therapeutically effective amount of herbal ingredients and; b. A therapeutically effective amount of cow urine (Gomutra).

2. The polyherbal anti-cancer formulation as claimed in claim 1, wherein the therapeutically effective amount of herbal ingredients comprising Zingibar officinale, Tinospora cordifolia, Swertia chirayita, Picrorhiza kurroa, Cissampelos pareiras, Cyperus otundus, Cedrus deodara, Marsdenia tenacissima, Holarrhena antidysenterica, and Hemidesmus indicus.

3. The polyherbal anti-cancer formulation as claimed in claim 1, wherein the therapeutically effective amount of herbal ingredients are used in the range of l-50mg, more preferably l-30mg and preferably l-20mg.

4. The polyherbal anti-cancer formulation as claimed in claim 1, wherein the therapeutically effective amount of cow urine is in the polyherbal anticancer formulation.

5. The polyherbal anti-cancer formulation as claimed in claim 1, wherein the phytochemicals present in the polyherbal anti-cancer formulation comprises of alkaloids, amino-acids, carbohydrates, flavonoids, cardiac glycoside, saponin glycoside, tannins, steroids and triterpenoids.

6. The polyherbal anti-cancer formulation as claimed in claim 1, wherein a polyherbal liquid formulation can be prepared from infusion, decoction, maceration, percolation, oil infusion and distillation process.

7. The process for preparation of a polyherbal anti-cancer formulationcomprises step: a. Weighing all herbal ingredients; b. Soaking all herbal ingredient in one liter cow urine for 8 hours; c. Adding equal quantity of cow urine into step (b) solution; d. Performing distillation of step (c) until all the liquid phase get distillate; e. Filtering the solution of step (d) to remove any foreign particles; f. Storing the solution of step (e).

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