|Title of Invention||
A PROCESS FOR PREPARATION OF A NOVEL HERBAL MEDICINAL COMPOSITION FOR CANCER TREATMENT FROM JANAKIA ARAYALPATHRA ROOT AND TRIHCOPUS ZEYLANICUS LEAF
|Abstract||1. A process for preparation of a herbal medicinal composition (phytomedicine) for cancer treatment from extracts of Janakia arayalpathra root and dried leaves of Trichopus zeylanicus in the ratio of 1: ■/ comprising of the following steps of i. collection of fresh leaves of Trichopus zeylanicus from the cultivated gardens (or wild habitat), drying and powdering in an ordinary mixer at low speed, ii. thoroughly mixing the extract of Janakia arayalpathra roots obtained, using methods, the dried leaf powder of Step I in the ratio of 1:1 using suspending agents like 2% gum acacia or 5% Tween 80 to obtain the herbal medicinal composition (phytomedicine) for cancer treatment.|
The present invention in general relates to a process for preparation of a herbal medicinal composition (formulation) for cancer treatment from the root of the plant Janakia arayalpathra, and the leaves of Thhcopus zeylanicus.
The cancer curative properties of the herbal formulation prepared from the above plants have been established by animal experiments. The formulation is prepared according to the Ayurvedic Pharmaceutical practices. The drug is free from any toxic side effects as evidenced from the toxicity studies.
The primary object of the invention Is to invent an effective cure for ascitic type of cancer.
It is another object of the present invention to Improve the efficiency of Janakia arayalpathra root extract as a cancer treating herbal drug by adding another medicinal plant Trichopus zeylanicus.
It is another object of the invention to invent a herbal drug, which will be effective and will not have any side effects.
The invention relates to a process for preparing a herbal formulation containing Janakia arayalpathra root and Trichopus zeylanicus leaf. The herbal medicine is a mixture of methanol extract of Janakia arayalpathra (mature root) and leaf powder of Trichopus zeylanicus subsp. travancoricus. This preparation (formulation) can effectively prevent the growth of Ehrlichs Ascitic Carcinoma (EAC) in mice.
One of the important aspects of the invention is the addition of Trichopus zeylanicus leaf powder in the anticancer extract from Janakia arayalpathra root, thereby dramatically improving the efficacy of the extract against EAC in mice. The formulation consisting of J- arayalpathra root extract and T. zeylanicus leaf powder, prepared by the process of this invention protected all the mice challenged vAth 1x10* EAC cells/mouse at a dose of 200 mg/kg (100 mg/kg J.arayalpathra root extract and 100 mg Tzeylanicus leaf powder).
Janakia arayalpathra Joseph and Chandrasekharan (Periplocaceae) is a rare and endemic undershrub found in Western Ghats, India. The tribals inhabiting the Western Ghat region of Kerala use the tuberous root of this plant for peptic ulcer and tumours of external organs (Pushpangadan P, Rajasekharan S, Ratheesh Kumar PK, Jawahar CR, Radhakrishnan K, Nair CPR, Sarada Amma L, Bhatt AV, 1990 Ancient Science of Life, 9, 13-15). Further, it has been shown that fresh Janakia arayalpathra root (1 gm/kg) has anti-EAC and immunomodulatory activities. The fresh root aqueous suspension (Igm/kg) protected about 50% of mice challenged with 0.5 x 10® EAC cells (Subramoniam A, Rajasekharan S, Latha PG, Evans DA and Pushpangadan P. 1996, Fitoterapia, 67, 140-144), Recent work carried out at TBGRI has shovm that ethanol extract of Jarjakia arayalpathra possesses anticancer activity. At an optimum dose (50 mg/kg) it protected 50% mice challenged with EAC cells and exhibited marked inhibition of solid (Daltons Lymphoma) tumour growth in mice. This anticancer activity of the extract was found to decrease when the concentration was increased above the optimum dose (50 mg/kg). However, at a lower dose (25 mg/kg) the herbal drug showed only marginal activity. A patent application has been filed on these observations by Department of Science and Technology, (Govt, of India). [Inventors: P. Pushpandagan, S.Rajasekharan, A. Subramoniam, V.George, P.G.Latha and V.V. Asha; File No.949/DEL/2000]. However, for effective treatment the potency of the drug has to be increased.
Trichopus zeylanicus is used by the Kani tribes of Agasthiar Hills of Kerala for getting instant stamina, better health and vitality. Earlier studies have shown that T. zeylanicus leaves have adaptogenic, aphrodisiac and antitoxic activities. The leaf powder (100 mg/kg) protected rats from paracetamol-overdose induced liver damage (Subramoniam, A, Evans DA, Rajasekharan S and Pushpangadan P, 1998, Indian J.Exptl. Bioi. 36,385-389). Limited single dose study has also shown that the plant leaf powder has some level of anti-EAC activity in mice
(Puspangadan P., Rajasekharan S, Subramoniam A, Latha PG, Evans DA and Valsaraj R (1995) Ancient Science of Life 54, 127-135).
The objective of the present invention is to improve the efficacy of Janakia atdyalpathra root extract with another medicinal (adaptogenic) plant. The same has been achieved.
STATEMENT OF INVENTION
A process for preparation of a herbal medicinal composition (phytomedicine) for cancer treatment from extracts of Janakia arayalpathra root and dried leaves of Trichopus zeylanicus in the ratio of 1: ? comprising of the following steps of
1) collection of fresh leaves of Triciiopus zeylanicus from the cultivated gardens (or wild habitat), drying and powdering in an ordinary mixer at low speed,
2) thoroughly mixing the extract of Janakia arayalpathra roots obtained, v/ith the dried leaf povi«jer of Step I in the ratio of 1:1 using suspending agents like 2% gum acacia or 5% Tween 80 to obtain the herbal medicinal composition (phytomedicine) for cancer treatment.
Detailed description of the process:
The mature leaves of Trichopus zeylanicus was collected from Tropical Botanic Garden and Research Institute and air-dried at (28-32 degree C) room temperature. The dried leaf was powdered in a mixie and the anticancer activity of the powder was tested using Ehrlich's Ascitic Carcinoma (EAC) in mice as a model. Swiss albino mice vrere challenged with EAC cells (half or one million EAC cells/mouse).
The leaf powder of Tnchopus zeyianicus was suspended in 2% gum acacia and administered orally to different groups at different doses (0, 50, 100 and 200 mg/kg) daily from the day of EAC cell challenge (1/2 million cells/mouse; i,p,); 10 mice were used in each group. The treatments were continued for 24 days or until the death of the animal. The peritonea! cavity of the surviving animals vras examined carefully after 40 days for the presence of tumour cells. The percentage protection to the tumour challenged mice provided by the drug is reflected in Figure. 1, Tnchopus zeyianicus leaf povwder shovred significant anti-EAC activity. At a dose of 100 or 200 mg/kg, the herbal drug protected 40-50% of EAC (0,5 X 106 cells/mouse) challenged mice. The animals which survived the cancer challenge did not show the presence of cancer cells in the peritoneal cavity, when examined. However, a lower dose (50 mg/kg) was devoid of anti-EAC activity (Figure 1).
When the mice were challenged with 1 million EAC cells/mouse instead of 0.5 million cells, 100 mg/kg Trichopus zeyianicus leaf administration did not provide protection from EAC (Fig. 2). This suggests that T, Zeyianicus can protect mice from EAC cells, when the cancer cells number is relatively less in the peritoneal cavity.
The mature roots of J.arayalpathra were collected, cleaned, cut into pieces and air dried at 28-32' C (room temperature) until constant weight was obtained. The dried material was powdered with a domestic mixer at low speed. The powdered material vras mixed separately vAih methanol or ethanol and the extract was dried and tested for its anticancer (Anti-EAC) activity. (Our previous study has shown that water as well as petroleum ether extracts are devoid of anticancer activity).
The powdered root of J. arayalpathra was mixed with alcohol (10 gm/IOOmI) and it was constantly stirred using a magnetic stirrer for eight hours and the filtrate was collected through Whatman No. 1 filter paper. The residue obtained was extracted again with fresh solvent for four hours and the filtrate was again collected. The filtrates collected as above were combined and evaporated to dryness using rotoevaporator to get it completely dried at 40'C, The powdered material was mixed separately with methanol or ethanoi and extracted under a pressure lower than atmospheric pressure. 100 gm of dried root powder yielded about 14 g of methanol extract. This extract was suspended in 5% Tv^reen 80 and used for the treatment. Swiss male albino mice (6-8 weeks old) were challenged with EAC (1 X 10° 6 cells/mouse; i.p.) and divided Into different groups of 10 mice each. One group of mice served as placebo control. The extract was administered (p.o) to different groups in indicated concentrations. (25, 50 or 100 mg/kg) daily from the day of tumour challenge. The treatments were continued for 24 days. The results are reflected in Figure 2. The methanol extract showed significant anti-cancer activity. At a dose of 50 mg/kg, the extract protected 50% of EAC (1x 10^ cells/mouse) challenged mice. The animal which survived the cancer challenge did not show the presence of cancer cells in the peritoneal cavity, when examined 40 days after tumour challenge. However, the methanol extract did not exhibit a concentration dependent anti-cancer activity. As shown in fig. 2 the maximum protection was observed at 50 mg/kg dose; at a higher dose (100 mg/kg) the activity was not found better than that at 50 mg/kg.
To improve the efficacy of J. arayalpathra root extracts, a herbal formulation (combination) vras prepared.
I. zeylanicus leaf powder was mixed with the methanol extract of J. arayalpathra root (1:1 ratio) to prepare the herbal combination. This formulation was suspended using 5% Tween 80 or 2% gum acacia as a suspending agent for oral administration.
This composition showed remarkable anti-EAC activity in mice. As shown in Figure 3, the combination protected all the mice which were challenged with 1 X 10°6 EAC cells at a dose of 200 mg/kg. All the untreated animals died of tumour burden within 20 days after tumour challenge. The drug treatment was discontinued after 24 days. When examined after 40 days, all the treated animals were found to be normal without any detectable EAC cells in the peritoneal cavity. Further the drug treatment did not result in any toxic symptoms in the mice. Thus it appears that this Is highly promising to treat at least certain types of cancer in the early stages of neoplastic growth.
To test whether the combination/formulation has anti-solid tumour activity, in addition to anti-EAC activity, the effect of the herbal formulation on the development of Daltons Lymphoma solid tumour in mice was done as described below.
Mice were challenged with Dalton's Lymphoma cells (10^ ceils/mouse, intramuscular injection) into the right hind limb to develop solid tumour. The mice were divided into 3 groups of 10 mice each. Group 1 was kept as placebo controls. Groups 3 and 2 received 100 and 200 mg/kg drug respectively, daily, (p.o) for 28 days. As given in Fig. 4 the solid tumour development was not substantially influenced by the administration of the herbal formulation; only marginal effect was observed. [T. zeylanicus powder is likely to promote new blood capillaries formation which is required to nourish growing solid tumour, whereas tumour cells in fluid do not require this. This is a possible reason for the ineffectiveness of the herbal combination in the case of DL-solid tumour].
The herbal fomiulation (the present invention) could be effective to cancer ceils in fluid state such as blood cancer, ascitic form of cancer, etc.
1. A process for preparation of a herbal medicinal composition (phytomedicine)
for cancer treatment from extracts of Janakia arayalpathra root and dried
leaves of Trichopus zeylanicus in the ratio of 1: ■/ comprising of the following
i. collection of fresh leaves of Trichopus zeylanicus from the cultivated gardens (or wild habitat), drying and powdering in an ordinary mixer at low speed,
ii. thoroughly mixing the extract of Janakia arayalpathra roots obtained, using methods, the dried leaf powder of Step I in the ratio of 1:1 using suspending agents like 2% gum acacia or 5% Tween 80 to obtain the herbal medicinal composition (phytomedicine) for cancer treatment.
2. The process as claimed in claim 1, Step II wherein the two ingredients i.e.
the extract of Janakia arayalpathra root and dried leaves of Trichopus
zeylanicus are mixed in the ratio of 1:1 (wt/wt ) using 5% Tween 80 as
3. The process for preparing a herbal medicinal composition (phytomedicine) as
described substantially herein.
|Indian Patent Application Number||659/MAS/2001|
|PG Journal Number||37/2010|
|Date of Filing||09-Aug-2001|
|Name of Patentee||TROPICAL BOTANIC GARDEN AND RESEARCH INSTITUTE|
|Applicant Address||KARIMANCODE P.O., PACKA-PALODE, THIRUVANANTHAURAM - 695562|
|PCT International Classification Number||461K35/68|
|PCT International Application Number||N/A|
|PCT International Filing date|