Cancer Research MAK-5 Amrit Kalash in Animal Models

Reduction of Metastases of Lewis Lung Carcinoma by an Ayurvedic Food Supplement [MAK-4] in Mice

Vimal K. Patel, PhD,* J. Wang, PhD,* R.N. Shen, MD,* H.M. Sharma, MD,***** and Z. Brahmi, PhD*

This study investigated the effect of oral feeding of an Ayurvedic rasayana (health-promoting/therapeutic herbal preparation) called Maharishi Arnrit

Kalash-4 (MAK-4) on metastasis of Lewis Lung Carcinoma (LLC) in mice. The mice were fed either chow containing 3% MAK-4 or standard laboratory chow, and inoculated subcutaneously with LLC tumor cells. After 4-3 weeks, the animals receiving the MAK-4-supplemented chow had a 65% reduction (p<0.01) in the

number of metastatic nodules, and a 43% reduction (p<0.01) in the size of the nodules, compared to the control group.

EXPT. 1 EXPT. 2 EXPT. 1 EXPT. 2

FIG. 1. Metastatic lung nodules (numbers/lung left side and

size > 4 mm/lung right side) in animals on M-4-containing chow

(Darkened Bars) and laboratory chow (Open Bars).

*P < .01 & *T < .001.

Abstract and figure reprinted bypermission of the publisher from Nutrition Research, Vol. 12, pp. 51-61. Copyright 1992 by

Elsevier Science Inc.

4. Title

Ayurvedic (Science of Life) Agents [MAK-4 and MAK-3] Induce Differentiation in Murine Neuroblastoma

Cells in Culture

Publication

Neuropharmacology, Vol. 31, No. 6, pp. 599-607, 1992.

Authors

K.N. Prasad,* Judith Edwards-Prasad,* Susan Kentroti,** C. Brodie,** and Antonia Vernadakis.**

Conducted at

* Center for Vitamins and Cancer Research, Department of Radiology, and

**Departments of Psychiatry and Pharmacology, University of Colorado Health Sciences Center,

Denver, CO 80262

Summary

This study shows that an ethanol extract of MAK-5 (also known as Maharishi Arnrit Kalash Ambrosia)

induced morphological differentiation (neurite formation) and biochemical differentiation (increased activity of tyrosine hydroxylase by about 13-fold) in 75% of murine neuroblastoma cells in culture (p<0.03), indicative of reversal of the malignant process. An aqueous extract of MAK-3 increased only the activity of tyrosine hydroxylase and to a lesser extent than the ethanol extract. A treatment time of 3 days was needed for the expression of maximum differentiation. Ethanol and aqueous extracts of MAK-3 also increased the intracellular level of adenosine 3 ',3 '-cyclic monophosphate (cAMP) by about 4-fold in 3 days. Ethanol extracts of MAK-5 also induced neurite formation in neuroblastoma cells grown in serum-free medium, but the concentration requirement was about a fifth of that needed in serum. A treatment time of 24 hours was sufficient to induce optimal differentiation in neuroblastoma cells grown in serum-free medium. The differentiating agents in the ethanol extract of MAK-5 were resistant to heat and light and could not be removed by treatment with activated charcoal. Neither the ethanol nor the aqueous extracts of MAK-4 (also known as Maharishi Arnrit Kalash Nectar) induced differentiation

Days After Treatment

Fig. 2. Effect of an ethanol extract of MAK.-A on morphological differentiation, as a function of treatment time. Ethanol-MAK-A (50 c gml) was added I day after plating. After 3 days of treatment, ethanol-MAK-A was removed and the number of morphologically differentiated cells was determined after I and 2 days of removal. Each value represents an average of 9 samples SEM. The sizes of bars

for the SEM at some points did not exceed the size of the symbol; therefore, they were not represented.

Table 2. Effects of extracts of Maharishi Arnril Kalish-Ambrosii (MAK-4) on activity of tyrosine hydroxylase in neuroblasloma cell in culture

Cells (0.25 x 10") for all groups, except those which receive 50 cg/ml ofethanol-MAK-A and 170 g/mlll ofaqueous-MAK 4 ; the latter were plated with I x 10* cells and were plated in tissue culture dishes (100 mm) and an ethanol extract an aqueous extract of MAK-A, were added separately 24 hr late: The medium and extracts were changed after 2 days of treatment and the activity of the enzyme was determined after 15 mm and 3 days of treatment. Each value represents an average of samples. Experiments were repeated 3 times and similar change were observed in the treated groups, in comparison to control; *Standard error of the mean. 'Significantly different (/* < 0.05) from control. *** Significantly different ( P < fl 05)

Level of cAMP

Table 3. Effects of extracts of Maharishi Amrit Kalish-Ambrosia (MAK-4) on the intracellular level of cAMP in neuroblasloma cell in culture (pmol/hr/mg protein)

Cells (50.000 cells for all groups except those which receive' 50 ip:ml and 170~g/ml of ethanol-MAK-4A. The latter were plated with 10" cells) plated in tissue culture dishes (60mm) in an ethanol extract and an aqueous extract of MAK.-4 were added separately 24 hr later. The medium and extracts were changed after 2 days of treatment and the level of cAMP was determined after 15min and 3 days of treatment. Each value represents an average of 3 samples. Experiments were repeated 3 limes and similar changes were observed in the treated group! in comparison to controls.

Abstract, figure and tables reprinted from Neuropharmacology, Vol. 31, No. 6, pp. 599-607, Copyright 1999.with permission from Elsevier Science Ltd, The Boulevard, Lang ford Lane, Kidlington 0X5 IGB, UK.