Patent Publication Number: US-2022233624-A9

Title: Anti-cancer and anti-proliferative compositions, and methods for their use in treating cancer

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application claims priority from Indian Provisional Application No. 202041009290 (IN), filed Mar. 4, 2020, which is incorporated by reference herein in its entirety. 
     FIELD OF THE INVENTION 
     The present invention relates to compositions and methods for treating cancer and/or inhibiting the proliferation of cancer cells and/or cancer-associated cells with compositions comprising  Terminalia chebula  (e.g. AyuFlex®),  Terminalia bellerica  (e.g. Ayuric®),  Phyllanthus emblica  (e.g. Capros®),  Withania somnifera  (e.g. Sensoril®), Shilajit (e.g. PrimaVie®),  Azadirachta indica  (e.g. PhytoBGS®), and combinations thereof, including for instance a trivalent chromium complex with extracts of Shilajit and  P. emblica  (e.g. Crominex-3+®). Combinations of the extracts, co-administration with anti-cancer drugs, and related methods are also described. 
     BACKGROUND 
       Terminalia chebula  has been extensively used in Ayurveda, Unani and Homoeopathic systems of medicine for improvement of different health conditions.  T. chebula  may be rich in tannoids and may contain a variety of other constituents. Chemical constituents isolated from  T. chebula  may vary considerably in type and/or concentration due to number of factors, e.g., ecological variation, soil variation, nutrient variation, as well as variations in the process of extraction. 
       Terminalia bellerica  (a.k.a. belerica, belirica, bellirica) is grown widely throughout India, Sri Lanka, and South East Asia.  T. bellerica  has been used for centuries in Ayurveda and may contain several chemical constituents in common with  T. chebula.    
       Phyllanthus emblica , the Indian gooseberry, is also widely used in Indian medicine for treatment of various diseases. 
       Withania somnifera , commonly known as Ashwagandha, has been used in herbal formulations of the Ayurvedic or Indian system of medicine, for instance to help to ward off stress and act as an adaptogen. 
     Shilajit, also known as “Moomiyo,” is found in high altitudes, for instance of the Himalayan Mountains, and is considered one of the “wonder medicines” of Ayurveda, the traditional Indian system of medicine dating back to 3500 B.C.E. Shilajit is physiologically active organic matter, composed of rock humus, rock minerals, and organic substances that have been compressed by layers of rock mixed with marine organisms and microbial metabolites. Shilajit oozes out of the rocks as a black mass in the Himalayas at higher altitudes ranging from 1000 to 5000 meters, as the rocks become warm during summer. Shilajit contains fulvic acids (“FAs”) as its main components, along with dibenzo-α-pyrones (“DBPs”) and DBP chromoproteins, humic acid, and more than forty (40) minerals. DBPs are also known as Urolithins (e.g. Urolithin A, Urolithin B). 
       Azadirachta indica , commonly known as neem, nitree, or Indian lilac, is a tree in the mahogany family Meliaceae. It is one of two species in the genus  Azadirachta , and is native to the Indian subcontinent, i.e., India, Nepal, Pakistan, Bangladesh, Sri Lanka, and Maldives. It is typically grown in tropical and semi-tropical regions. Neem trees also grow in islands located in the southern part of Iran. Its fruits and seeds are the source of neem oil. 
     Products made from neem trees have been used in India for over two millennia for their medicinal properties. Neem products are considered a major component in Siddha medicine and Ayurvedic and Unani medicine and are particularly prescribed for skin diseases. Neem oil is also used to promote healthy hair, to improve liver function, detoxify the blood, and balance blood sugar levels. Neem leaves have also been used to treat skin diseases like eczema and psoriasis. 
     Primary patient-derived cancer cells (PDCs), cultured directly from tissue obtained from surgery in cancer patients, provide a physiologically relevant platform for testing an individual&#39;s cancer cells and cell type responses to a specific drug or composition. Information derived from PDC testing may be useful in personalized as well as generalized cancer patient treatments. 
     In addition, in-vitro studies using standardized cancer cell lines provide valuable information for animal and human treatments. 
     SUMMARY OF THE INVENTION 
     The present invention is directed to a method of inhibiting the proliferation of cancer cells and/or cancer-associated cells comprising the steps of providing a composition comprising  Terminalia chebula  (for instance, AyuFlex®),  Terminalia bellerica  (for instance, Ayuric®),  Phyllanthus emblica  (for instance, Capros®),  Withania somnifera  (for instance, Sensoril®), Shilajit (for instance, PrimaVie®), and/or  Azadirachta indica  (for instance, PhytoBGS®), and combinations thereof, including for instance a trivalent chromium complex with extracts of Shilajit and  P. emblica  (e.g. Crominex-3+®), and applying the composition to cancer cells in an amount effective to cause an anti-proliferative effect. In an embodiment, cancer cells inhibited according to the present invention include glioma cells, breast cancer cells (ER/PR+ Her2 equivocal (“HR+”), ER/PR− Her2+(“Her2+”), and Triple Negative (“TN”)), chronic lymphocytic leukemia cells (“CLL”), acute myeloid leukemia cells (“AML”), small cell lung cancer cells, non-small lung cancer cells, colon cancer cells, pancreatic cancer cells, prostate cancer cells, and/or ovarian cancer cells. 
     Also, the present invention is directed to a method of treating cancer in a subject in need of such treatment, comprising the steps of providing a composition comprising  Terminalia chebula  (for instance, AyuFlex®),  Terminalia bellerica  (for instance, Ayuric®),  Phyllanthus emblica  (for instance, Capros®),  Withania somnifera  (for instance, Sensoril®), Shilajit (for instance, PrimaVie®), and/or  Azadirachta indica  (for instance, PhytoBGS®), and combinations thereof, including for instance a trivalent chromium complex with extracts of Shilajit and  P. emblica  (e.g. Crominex-3+®), and administering the composition to the subject in an amount effective to treat the cancer, for instance by slowing the progression or growth of the cancer, stopping the progression or growth of the cancer, shrinking the cancerous tumor, reducing the number of cancer cells and/or cancer-associated cells in the subject, and/or rendering the cancer almost undetectable or undetectable in the subject. In an embodiment, a cancer treated according to the present application includes glioma, breast cancer (including ER/PR+ Her2 equivocal, ER/PR− Her2+, and Triple Negative), chronic lymphocytic leukemia, acute myeloid leukemia, colon cancer, small-cell lung cancer, non-small cell lung cancer, pancreatic cancer, prostate cancer, and/or ovarian cancer. In an embodiment, the composition is an extract, for instance a standardized aqueous extract, of the plants identified above. In an embodiment, a method of treatment according to this invention includes administering compositions such as extracts of the above with anti-cancer drugs to enhance the effects of the drugs and, in an embodiment, reduce side, adverse, or toxic effects of the drugs, in an embodiment by reducing the necessary dose of the traditional drug. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a graph illustrating anti-proliferative and anti-cancer effects of AyuFlex® on cancer PDCs (left to right: Glioma, Breast Cancer (HR+), Breast Cancer (Her2+), Breast Cancer (TN), CLL, AML) in an anti-proliferation assay (“% INHIBITION” marked at 0, 25, 50, 75, 100%, with extract concentrations shown in 3 concentrations for each cancer type, left to right: 100 ug/ml, 30 ug/ml, 10 ug/ml). 
         FIG. 2  is a graph illustrating anti-proliferative and anti-cancer effects of Ayuric® on cancer PDCs (left to right: Glioma, Breast Cancer (HR+), Breast Cancer (Her2+), Breast Cancer (TN), CLL, AML) in an anti-proliferation assay (“% INHIBITION” marked at 0, 25, 50, 75, 100%, with extract concentrations shown in 3 concentrations for each cancer type, left to right: 100 ug/ml, 30 ug/ml, 10 ug/ml). 
         FIG. 3  is a graph illustrating anti-proliferative and anti-cancer effects of Capros® on cancer PDCs (left to right: Glioma, Breast Cancer (HR+), Breast Cancer (Her2+), Breast Cancer (TN), CLL, AML) in an anti-proliferation assay (“% INHIBITION” marked at 0, 25, 50, 75, 100%, with extract concentrations shown in 3 concentrations for each cancer type, left to right: 100 ug/ml, 30 ug/ml, 10 ug/ml). 
         FIG. 4  is a graph illustrating anti-proliferative and anti-cancer effects of Sensoril® on cancer PDCs (left to right: Glioma, Breast Cancer (HR+), Breast Cancer (Her2+), Breast Cancer (TN), CLL, AML) in an anti-proliferation assay (“% INHIBITION” marked at 0, 25, 50, 75, 100%, with extract concentrations shown in 3 concentrations for each cancer type, left to right: 100 ug/ml, 30 ug/ml, 10 ug/ml). 
         FIG. 5  is a graph illustrating anti-proliferative and anti-cancer effects of PrimaVie® Shilajit on cancer PDCs (left to right: Glioma, Breast Cancer (HR+), Breast Cancer (Her2+), Breast Cancer (TN), CLL, AML) in an anti-proliferation assay (“% INHIBITION” marked at 0, 25, 50, 75, 100%, with extract concentrations shown in 3 concentrations for each cancer type, left to right: 100 ug/ml, 30 ug/ml, 10 ug/ml). 
         FIG. 6  is a graph illustrating anti-proliferative and anti-cancer effects of PhytoBGS® on cancer PDCs in an anti-proliferation assay (“% INHIBITION” marked at 0, 25, 50, 75, 100%, with extract concentrations shown in 3 concentrations for each cancer type, left to right: 100 ug/ml, 30 ug/ml, 10 ug/ml). 
         FIG. 7  is a graph illustrating anti-proliferative and anti-cancer effects of AyuFlex®, Ayuric®, Capros®, Sensoril®, PrimaVie®, and PhytoBGS® on glioma PDCs in an anti-proliferation assay. 
         FIG. 8  is a graph illustrating the efficacy of Ayuflex®, Capros®, and Ayuric® on glioma cancer cells in an anti-proliferation assay, alone or in combination with temozolomide, or in the combinations Ayuflex®+Capros®, Ayuflex®+Ayuric®, Capros®+Ayuric®. 
         FIG. 9  is a graph illustrating anti-proliferative and anti-cancer effects of AyuFlex®, Ayuric®, Capros®, Sensoril®, PrimaVie®, and PhytoBGS® on breast cancer “HR+” PDCs in an anti-proliferation assay. 
         FIG. 10  is a graph representing a dose-response curve of Ayuflex® on HR+ breast cancer cells. 
         FIG. 11  is a graph illustrating the efficacy of Ayuflex®, Capros®, and Ayuric® on HR+ breast cancer cells in an anti-proliferation assay, each in combination with docetaxel or 5-fluorouracil (5-FU), or in the combinations Ayuflex®+Capros®, Ayuflex®+Ayuric®, Capros®+Ayuric®. 
         FIG. 12  is a graph illustrating the efficacy of Ayuflex®, Capros®, and Ayuric® on HR+ breast cancer cells in an anti-proliferation assay, alone or in combination with docetaxel or 5-fluorouracil (5-FU), or in the combinations Ayuflex®+Capros®, Ayuflex®+Ayuric®, Capros®+Ayuric®, grouped by extract according to the invention. 
         FIG. 13  is a graph illustrating anti-proliferative and anti-cancer effects of AyuFlex®, Ayuric®, Capros®, Sensoril®, PrimaVie®, and PhytoBGS® on breast cancer “Her2+” PDCs in an anti-proliferation assay. 
         FIG. 14  is a graph representing a dose-response curve of Ayuflex® on Her2+ breast cancer cells. 
         FIG. 15  is a graph illustrating the efficacy of Ayuflex®, Capros®, and Ayuric® on Her2+ breast cancer cells in an anti-proliferation assay, each in combination with docetaxel or 5-fluorouracil, or in the combinations Ayuflex®+Capros®, Ayuflex®+Ayuric®, Capros®+Ayuric®. 
         FIG. 16  is a graph illustrating anti-proliferative and anti-cancer effects of AyuFlex®, Ayuric®, Capros®, Sensoril®, PrimaVie®, and PhytoBGS® on breast cancer “TN” (Triple Negative) PDCs in an anti-proliferation assay. 
         FIG. 17  is a graph representing a dose-response curve of Ayuflex® on TN breast cancer cells. 
         FIG. 18  is a graph illustrating the efficacy of Ayuflex®, Capros®, and Ayuric® on TN breast cancer cells in an anti-proliferation assay, each in combination with docetaxel or 5-fluorouracil, or in the combinations Ayuflex®+Capros®, Ayuflex®+Ayuric®, Capros®+Ayuric®. 
         FIG. 19  is a graph illustrating anti-proliferative and anti-cancer effects of AyuFlex®, Ayuric®, Capros®, Sensoril®, PrimaVie®, and PhytoBGS® on CLL (Chronic lymphocytic leukemia) PDCs in an anti-proliferation assay. 
         FIG. 20  is a graph representing a dose-response curve of Ayuflex®, Capros®, Ayuric®, and the drug ibrutinib on CLL cancer cells. 
         FIG. 21  is a graph illustrating the efficacy of Ayuflex®, Capros®, and Ayuric® on CLL cancer cells in an anti-proliferation assay, alone or in combination with ibrutinib, or in the combinations Ayuflex®+Capros®, Ayuflex®+Ayuric®, Capros®+Ayuric®. 
         FIG. 22  is a graph illustrating anti-proliferative and anti-cancer effects of AyuFlex®, Ayuric®, Capros®, Sensoril®, PrimaVie®, and PhytoBGS® on AML (Acute myeloid leukemia) PDCs in an anti-proliferation assay. 
         FIG. 23  is a graph representing a dose-response curve of PhytoBGS®, PrimaVie®, and Sensoril® on AML cancer cells. 
         FIG. 24  is a graph illustrating the efficacy of Phyto-BGS®, PrimaVie®, and Sensoril® on AML cancer cells in an anti-proliferation assay, alone or in combination with arsenic trioxide, cytarabine, or doxorubicin, or in the combinations PhytoBGS®+PrimaVie®, PhytoBGS®+Sensoril®, and PrimaVie®+Sensoril®. 
         FIG. 25  is a graph representing a dose-response curve of Sensoril® on AML subtype Acute Promyelocytic Leukemia (APL, APML) patient-derived cancer cells (PDCs), showing a 50% Inhibitory Concentration (IC 50 ) of 32.94 ug/ml. 
         FIG. 26  is a graph showing the inhibition of APL PDC proliferation with Sensoril® and cytarabine. 
         FIG. 27  is a graph showing the inhibition of APL PDC proliferation with Sensoril® alone and synergistically with Sensoril® and arsenic trioxide (As 2 O 3 ) in combination. 
         FIG. 28  is a graph showing the inhibition of APL PDC proliferation with Sensoril® alone and synergistically with Sensoril® and doxorubicin in combination. 
         FIG. 29  is a graph showing the inhibition of non-small cell lung cancer cell proliferation with extracts of this invention. 
         FIG. 30  is a graph showing the inhibition of colon cancer cell proliferation with extracts of this invention. 
         FIG. 31  is a graph illustrating the efficacy of Ayuflex®, Capros®, and Ayuric® on glioma PDCs SB 32833 (Glioblastoma Grade IV) in an anti-proliferation assay, alone or in the combinations Ayuflex®+Capros®, Ayuflex®+Ayuric®, Capros®+Ayuric®, with GDC-0941 (10 uM), Doxorubicin (10 uM), and Temozolomide (100 uM) as controls. 
         FIG. 32  is a graph illustrating the efficacy of Ayuflex®, Capros®, and Ayuric® on glioma SB 6129 (anaplastic astrocytoma Grade III) patient-derived cancer cells in an anti-proliferation assay, alone (left to right: AyuFlex, Capros, Ayuric, or in combination Ayuflex®+Capros®, Ayuflex®+Ayuric®, Capros®+Ayuric®, with GDC-0941 (10 uM), Doxorubicin (10 uM), and Temozolomide (100 uM) as controls. 
         FIG. 33  is a graph illustrating the efficacy of extracts of this invention in inhibiting proliferation of glioblastoma cells (U87-MG cell line). 
         FIG. 34  is a graph illustrating the efficacy of extracts of this invention in inhibiting proliferation of glioblastoma cells (U87-MG cell line). 
         FIG. 35  illustrates dose-response curves of inhibitory activity of Sensoril® (IC 50  67 ug/ml), cytarabine (IC 50  567 nM), As 2 O 3  (IC 50  2.3 uM), and doxorubicin (IC 50  79 nM) on AML cells (HL60 cell line). 
         FIG. 36  is a graph illustrating the efficacy of extracts of this invention in inhibiting Triple Negative Breast Cancer cell proliferation (SB 30750 PDCs) after a 72-hour incubation period. 
         FIG. 37  is a graph illustrating the efficacy of extracts of this invention in inhibiting Triple Negative Breast Cancer cell proliferation (MDAMB-231 cell line) after a 72-hour incubation period. 
         FIG. 38  is a graph illustrating the efficacy of extracts of this invention in inhibiting Triple Negative Breast Cancer cell proliferation (MDAMB-231 cell line) after a 120-hour incubation period. 
         FIG. 39  is a graph comparing the efficacy of extracts of this invention in inhibiting MDAMB-231 Triple Negative Breast Cancer cell proliferation (MDAMB-231 cell line) after 72- and 120-hour incubation periods. 
         FIG. 40  is a graph illustrating the efficacy of AyuFlex®, Ayuric®, and other extracts of this invention in inhibiting proliferation of small cell lung cancer cells. Extract concentrations are, left to right, 100 ug/ml, 30 ug/ml, 10 ug/ml. 
         FIG. 41  is a graph illustrating the efficacy of Ayuric®, Ayuflex®, Capros®, and other extracts of this invention in inhibiting proliferation of prostate cancer cells. Extract concentrations are, left to right, 100 ug/ml, 30 ug/ml, 10 ug/ml. 
         FIG. 42  is a graph illustrating the efficacy of Ayuflex®, Ayuric®, Sensoril®, and other extracts of this invention in inhibiting proliferation of ovarian cancer cells. Extract concentrations are, left to right, 100 ug/ml, 30 ug/ml, 10 ug/ml. 
         FIG. 43  illustrates dose-response curves of inhibitory activity of 6 different samples of hydroethanolic extracts of  Withania somnifera  on AML HL60 cell line cancer cells, and provides an IC 50  for each sample. 
         FIG. 44  is a graph illustrating the efficacy of standardized aqueous and hydroethanolic extracts of  Withania somnifera  (Sensoril®), Ayuflex®, Ayuric®, and other extracts of this invention in inhibiting proliferation of histiocytic lymphoma cancer cells. Extract concentrations are, left to right, 100 ug/ml, 30 ug/ml, 10 ug/ml. 
         FIG. 45  is a graph illustrating the efficacy of standardized aqueous and hydroethanolic extracts of  Withania somnifera  (Sensoril®), Ayuflex®, and other extracts of this invention in inhibiting proliferation of pancreatic cancer cells. Extract concentrations are, left to right, 100 ug/ml, 30 ug/ml, 10 ug/ml. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The below definitions and discussion are intended to guide understanding but are not intended to be limiting with regard to other disclosures in this application. References to percentage (%) in compositions of the present invention refers to the % by weight of a given component to the total weight of the composition being discussed, also signified by “w/w”, unless stated otherwise. 
     In the present invention, “anti-cancer” generally refers to preventing, treating and/or otherwise halting cancer in a subject for instance by reducing viability of cancer cells and/or cancer-associated cells, including for instance slowing the progression or growth of the cancer, stopping the progression or growth of the cancer, shrinking the cancerous tumor, reducing the number of cancer cells and/or cancer-associated cells in the subject, and/or rendering the cancer almost undetectable or undetectable in the subject. In an embodiment, cancer in a subject is diagnosed by a medical provider. In an embodiment, cancer in a subject is not diagnosed by a medical provider. 
     In the present invention, “anti-proliferative” and the like refers to inhibiting the proliferation of cancer cells, including for instance inhibiting the proliferation of cancer cells in a subject or for instance inhibiting the proliferation of cancer cells such as PDCs in a cell culture outside of a subject&#39;s body, by rendering the cancer cells temporarily or permanently non-viable (not able to grow or develop) and metabolically inactive. Reference to cells “inhibited by” a composition of this invention, or other substance, is to the inhibition of proliferation of cancer cells by the composition, for instance as shown by the anti-proliferation assays described in the Examples below, unless stated otherwise. In an embodiment, inhibiting the proliferation of cancer cells refers to killing cancer cells. In an embodiment, the proliferation of cancer cells may be inhibited by a composition of the present invention by 1%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or for instance by 1%-100%, or any range of numbers therein, such as for instance 33-37% or 62%-75%. In an embodiment, the number of viable cancer cells may be reduced by 1%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, by 1%-100%, or any range of numbers therein, such as for instance 2-8% or 79-93%. Known anti-cancer agents GDC-0941 (a class I phosphatidylinositol 3 kinase (PI3K) inhibitor; pictilisib) and doxorubicin are shown as anti-proliferative drug controls in Example I below, inhibiting the proliferation of cancer cells by about 50% to about 100%, depending e.g. on the sensitivity of the PDC type. An “anti-cancer drug” may be pictilisib, doxorubicin, temozolomide, docetaxel, 5-fluorouracil (5-FU), ibrutinib, arsenic trioxide, and/or cytarabine, and/or any other control or standard of care drug identified in this application. Comparisons of different concentrations of standardized aqueous extracts of the present invention with the inhibition of proliferation of PDCs by GDC-0941 and doxorubicin are also shown in the Figures. 
     In the present invention, “cancer cells” refers to cells of, or taken from or derived from, a cancerous source, such as a solid cancerous tumor or hematopoietic cancerous tissue or cells. An example of “cancer cells” of the present invention is a mixed culture of primary patient-derived cancer cells (PDC), taken directly from an individual human subject&#39;s cancerous tissue post-surgery. PDCs may include cells from any cancer, including for instance cells from a glioma, such as a glioblastoma Grade IV; cells from a breast cancer tumor, such as breast cancer cells that are ER/PR+ Her2 equivocal, ER/PR-Her2+, or triple negative; leukemia cells from a subject having chronic lymphocytic leukemia; and/or leukemia cells from a subject having acute myeloid leukemia (e.g. M 4 subtype), and so forth. See for instance Table 1. Also, cancer cells of this invention include cells of a standard cell line, such as the cell lines discussed in the below Examples. In an embodiment, cancer cells of the present invention are part of a tumor or other cancerous cell source in a mammalian subject, such as in a human body. In an embodiment, cancer cells of the present invention include characteristics attributed to cells of a cancerous tumor or malignancy or the like, for instance as known in the art. 
     In the present invention, “applying” (and the like) a composition of the present invention refers to making the composition physically available to the cancer cells, for instance by administering the extract to a subject having cancer so that the extract or at least its active components reach the cancer cells in the subject&#39;s body. Applying the extract in an “effective amount” to cancer cells refers to applying the extract in an amount that will inhibit proliferation of the cancer cells, for instance as described in Example I and other Examples below. 
     In the present invention, “administering”, “administration”, and the like refer to providing a composition of the present invention to a subject so that the composition (or components thereof) reaches the subject&#39;s bloodstream and/or tissues and thus reaches cancer cells, and acts on the cancer cells to slow and/or stop their proliferation and render them non-viable. Administration may be by the subject or by another. Administration to the subject may be oral, for instance in the form of a dietary supplement, and/or in a solid pharmaceutical dosage form, preferably in a discrete dose unit, such as a table, hard gelatin capsule, soft gelatin capsule, etc. Administration may also be through parenteral, intramuscular, transdermal, topical, sublingual, intravenous, and other physiologically acceptable routes. 
     In an embodiment, references to active components and the like throughout this application are not intended as being bound by theory, as it is the administration of the full composition of the present invention which is shown to provide the remarkable inhibition of cancer cells of the present invention. 
     “Co-administration” refers to administering two or more compositions such as extract(s) of the present invention, or one or more such compositions with another composition or with an anti-cancer drug. Such co-administration may be at different times, so long as both extract and anti-cancer drug are available in the bloodstream and/or tissues of the subject in an effective amount to slow and/or stop the proliferation of cancer cells and in an embodiment render them non-viable. Doses of anti-cancer drug may be per the normal standard of care or at the lower end or in a lower dose, for instance in view of a subject&#39;s intolerance of adverse effects from the medication. 
     In the present invention, “treating cancer” and the like refers to halting the progression of cancer in a subject and/or causing the cancer including e.g. cancer cells to decrease, shrink, and if possible become or remain undetectable in the subject, for instance by reducing the size of a cancer tumor and/or number of cancerous cells. In an embodiment, the number of viable cancer cells and/or size of the tumor may be reduced by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, by 1%-100%, or any range of numbers therein. In an embodiment, cancer being treated according to this invention is diagnosed in a subject by a medical professional, such that the subject has been diagnosed as having cancer. In an embodiment, cancer being treated according to this invention has not been diagnosed in a subject by a medical professional, such that the subject has not been diagnosed as having cancer, however cancer cells exist in the subject. 
     In an embodiment, a “subject” according to this invention is a human; in another embodiment, the subject is a mouse, rat, dog, horse, or any mammal including for instance a mammal serving as a model for cancer research, or a human. In an embodiment, treating the cancer with an “effective amount” of a composition of this invention includes administering an amount effective to slow the progression or growth of the cancer, stop the progression or growth of the cancer, shrink the cancerous tumor, reduce the number of cancer cells in the subject, and/or render the cancer almost undetectable or undetectable in the subject, and/or act against the cancer for instance as discussed elsewhere in this application or in the state of the art. “Enhancing” the treatment of cancer refers to administering a composition of this invention to a subject undergoing treatment with a different anti-cancer drug, to improve treatment outcome, as discussed throughout this application. 
     Compositions 
     A “composition” of the present invention comprises  Terminalia chebula  (e.g. AyuFlex®),  Terminalia bellerica  (e.g. Ayuric®),  Phyllanthus emblica  (e.g. Capros®),  Withania somnifera  (e.g. Sensoril®), Shilajit (e.g. PrimaVie®),  Azadirachta indica  (e.g. PhytoBGS®), or a combination thereof, including for instance a trivalent chromium complex with extracts of Shilajit and  P. emblica  (e.g. Crominex-3+®). In an embodiment, a composition consists essentially of, or consists of, one or more of the above. 
     In an embodiment, a composition of this invention is an extract of  Terminalia chebula  (e.g. AyuFlex®),  Terminalia bellerica  (e.g. Ayuric®),  Phyllanthus emblica  (e.g. Capros®),  Withania somnifera  (e.g. Sensoril®), Shilajit (e.g. PrimaVie®),  Azadirachta indica  (e.g. PhytoBGS®), or a combination thereof, including for instance a trivalent chromium complex with extracts of Shilajit and  P. emblica  (e.g. Crominex-3+®). In an embodiment, an extract of this invention refers to a natural substance prepared from  Terminalia chebula  fruit,  Terminalia bellerica  fruit,  Phyllanthus emblica  fruit,  Withania somnifera  roots and leaves, Shilajit,  Azadirachta indica  twigs and leaves, or a combination thereof, that has been disrupted from its natural state and treated with water or aqueous solution such as phosphate buffered saline (PBS), alcohol such as methanol or ethanol, or a hydroalcoholic mixture. In an embodiment a hydroalcoholic mixture may include alcohol and water together, for instance in a ratio of 9:1 to 1:9; in another embodiment, an extract may be prepared by first treating the plant or e.g. herbomineral material with one substance, such as ethanol, and then with a second substance, such as water or an aqueous solution. See for instance Example IX. In an embodiment, extracted substances may be pooled. In an embodiment, an extract of this invention is a standardized extract. 
     In an embodiment, an aqueous standardized extract of this invention is AyuFlex®, Ayuric®, Capros®, Sensoril®, PrimaVie®, and/or PhytoBGS®. A composition of the present invention may be a combination, for instance, a blend of one or more extracts such as AyuFlex®, Ayuric®, Capros®, Sensoril®, PrimaVie®, and/or PhytoBGS®, and/or a trivalent chromium complex such as Crominex+3®. In an embodiment Ayuflex® and Ayuric® are combined. In another embodiment, AyuFlex® and Capros® are combined. In an embodiment, Ayuric® and Capros® are combined. In an embodiment, AyuFlex®, Ayuric®, and Capros® are all combined in a composition of the present invention. In an embodiment, Sensoril® and PrimaVie® are combined. In an embodiment, Sensoril® and Phyto-BGS® are combined. In an embodiment, PrimaVie® and Phyto-BGS® are combined. In an embodiment, Sensoril®, PrimaVie®, and Phyto-BGS® are combined. Other combinations of the present extracts may be combined into a composition of the present invention. A composition of the present invention may comprise, consist essentially of, or consist of, any extract of the present invention or combination thereof. 
     In an embodiment, an extract such as an aqueous standardized extract is in powdered form and may be blended together with another extract or trivalent chromium complex such as Crominex+3® or another substance, for instance in powdered form, or other solid or form. In another embodiment, the extract(s) may be for instance blended or dissolved into a composition in liquid form, or combined in another form. A composition of the present invention may further comprise one or more excipients, additives, and/or other substances, including for instance microcrystalline cellulose, croscarmellose sodium, magnesium stearate, and/or silicon dioxide; and/or a suitable aqueous solution such as a buffer solution. A composition of the present invention may be formulated into nutraceutical or pharmaceutical dosage forms comprising for instance tablets, capsules, powders, liquids, chews, gummies, transdermals, injectables, dietary supplements, topical creams or gels, lozenges, pills, and so forth. In an embodiment, a composition of the present invention is a solution comprising an extract and applied to cancer cells as in the Examples below, or used to prepare the applied solutions. 
     In an embodiment, a composition comprising one or more standardized aqueous extracts of this invention is administered in an effective amount to a mammal, including a daily dose for a human being of at least 1-2000 mg of at least one extract, in an embodiment at least 50 mg, 100 mg, 250 mg, 500 mg, 750 mg, 1000 mg, 1500 mg, or 2000 mg; and optionally 1-20000 mg of each and any other extract included in the composition. In a mammal, a dose may be about the same as in a human, and may be adjusted per kilogram of weight of the mammal. A powdered blend of one or more extracts and optionally excipients or other substances such as fillers, disintegrants, flow enhancers, and lubricants, for instance, microcrystalline cellulose, croscarmellose sodium, silicon dioxide, and magnesium stearate, may be blended using standard powder blending techniques. In an embodiment, a composition of this invention may be marked as organic by an appropriate agency or organization. 
     Bioactive Components of Extracts of this Invention 
     AyuFlex® of this invention is a standardized aqueous extract of the fruits of  Terminalia chebula  plant, off white to brown color powder and soluble in water with astringent taste. It contains not less than 39% w/w low molecular weight hydrolysable tannins as bioactives with not less than 27% w/w chebulinic acid and chebulagic acid combined and not less than 12% w/w of other unindentified low molecular weight hydrolysable tannins. Ellagic acid and gallic acid are also present in the extract, for which the analytical results may be reported without any specification, but specifications for these bioactives may also be identified, for instance in an embodiment, each may be present in amounts of less than 10% of the extract or composition. In an embodiment, an aqueous extract of this invention, Ayuflex®, contains about 65-70% w/w low molecular weight hydrolysable tannins including about 45-50% w/w chebulinic acid and chebulagic acid combined. In an embodiment, said aqueous extract may have a measured amount of 8-9% each of gallic acid and ellagic acid. In an embodiment, a composition of this invention may be a preparation of  Terminalia chebula  fruit, dried and powdered and in an embodiment, standardized for pharmaceutical or nutraceutical usage. In an embodiment, an extract of this invention may be prepared from such a fruit preparation. 
       Terminalia bellerica  (for instance, Ayuric®)—Ayuric® of this invention is a standardized aqueous extract of the fruits of  Terminalia bellerica  plant, brown color powder and soluble in water with astringent taste. It contains not less than 15% w/w of low molecular weight hydrolysable tannins as bioactives, including chebulinic acid and chebulagic acid and other unidentified low molecular weight hydrolysable tannins. Ellagic acid and gallic acid are also present in the extract, for which the analytical results are only reported without any specification, but specifications for these bioactives may also be identified. In an embodiment, Ayuric® is a composition of this invention and contains 33-38% w/w low molecular weight hydrolysable tannins, including chebulinic acid and chebulagic acid and other unidentified low molecular weight hydrolysable tannins, and in a further embodiment, also contains 6-8% w/w gallic acid and 0.5-2% w/w ellagic acid. In an embodiment, a composition of this invention may be a preparation of  Terminalia bellerica  fruit, dried and powdered and in an embodiment, standardized for pharmaceutical or nutraceutical usage. In an embodiment, an extract of this invention may be prepared from such a fruit preparation. 
       Phyllanthus emblica  (for instance, Capros®)—Capros® of this invention is a standardized aqueous extract of the fruits of  Phyllanthus emblica  plant, light yellow color powder and soluble in water with astringent taste. It contains not less than 60% w/w of low molecular weight hydrolysable tannins, comprising of emblicanin-A, emblicanin-B, punigluconin and pedunculagin, and not more than 4% w/w of gallic acid. In an embodiment, Capros® as a composition of this invention contains about 75-80% w/w low molecular-weight hydrolysable tannins, and about 0.5-1.5% w/w gallic acid. In an embodiment, a composition of this invention may be a preparation of  Phyllanthus emblica  fruit, dried and powdered and in an embodiment, standardized for pharmaceutical or nutraceutical usage. In an embodiment, an extract of this invention may be prepared from such a fruit preparation. 
       Withania somnifera  (for instance, Sensoril®)—Sensoril® of this invention is a standardized aqueous extract of the roots and leaves of  Withania somnifera  plant, brown color powder and soluble in water with bitter taste. It contains not less than 10% w/w of withanolide glycosides, less than 0.5% w/w of withanolide aglycones (as Withaferin-A) and not less than 32% w/w oligosaccharides as bioactives. Withanolides, such as withastromolide, withanolide A, withanolide B, 27-hydroxy withanone, withanone, etc. may also be present in this extract. In an embodiment, Sensoril® as a composition of this invention contains about 10-12% w/w of withanolide glycosides, about 0-0.5% w/w of withanolide aglycones (as Withaferin-A), and 35-40% w/w oligosaccharides. 
     In an embodiment of the present invention, an extract of  Withania somnifera  is made by using ethanol extraction followed by water extraction. This extract is referred to as Sensoril®-AWE (“Alcohol-Water-Extract”). Other alcohols, such as methanol, isopropyl alcohol, etc. may also be used as extraction solvents instead of ethanol. Such a hydro-alcoholic Sensoril®-AWE extract of this invention contains not less than 10% w/w withanolide glycosides, not less than 2%, preferably not less than 3.0% w/w withanolide aglycones (as Withaferin-A) and not less than 20% w/w oligosaccharides as bioactives. Withanolides, such as withastromolide, withanolide A, withanolide B, 27-hydroxy withanone, withanone, etc. may also be present in this extract. In an embodiment, a composition of this invention is a hydroethanolic  Withania somnifera  (Sensoril®-AWE) extract containing 12-20% w/w withanolide glycosides (including as mentioned elsewhere in this application ranges therein, such as 12% w/w, 13% w/w, and so on), 2-8% w/w withanolide aglycones (as Withaferin-A), and 24-35% w/w oligosaccharides. 
     Shilajit (for instance, PrimaVie®)—PrimaVie® of this invention is a standardized aqueous extract of Shilajit, an herbo-mineral exudate from the Himalayan, Altai and other mountains, brown color powder and soluble in water with earthy taste. It contains not less than 50% w/w of fulvic acid and not less than 10.3% w/w of free dibenzo-α-pyrones and dibenzo-α-pyrones conjugated with chromoproteins combined. Dibenzo-α-pyrones are also known as Urolithins, for example Urolithin A and Urolithin B. In an embodiment, PrimaVie® as a composition of this invention is a standardized aqueous extract containing 58-63% fulvic acids with a dibenzo pyrone core nucleus and 14-18% of free dibenzo-α-pyrones and dibenzo-α-pyrones conjugated with chromoproteins combined. 
       Azadirachta indica  (for instance, PhytoBGS®)—PhytoBGS® of this invention is a standardized aqueous extract of the leaves and twigs of  Azadirachta indica  plant, brown color powder and soluble in water with bitter taste. It contains not less than 2% w/w of flavonoids (comprising of quercetin-3-O-glucoside, quercetin-3-O-rutinoside, apigenin rutionoside and other rutin derivatives) and not less than 5% w/w and up to 20% w/w of myoinositol monophosphate as bioactives, and is devoid of possibly toxic compounds such as azadirachtone, azadiradione, nimbolide, nimbin. In an embodiment, a PhytoBGS® composition of this invention contains about 3% w/w flavonoids and 6-8% w/w myoinositol monophosphate. 
     Trivalent chromium complex (for instance, Crominex-3+®)—Crominex-3+® of this invention is a complex of trivalent chromium chloride,  Phyllanthus emblica  extract and Shilajit extract. It is a light brown color powder, soluble in water with astringent taste. It contains not less than 2% w/w of trivalent chromium as the bioactive. In an embodiment, a composition of this invention is Crominex-3+® containing 2-3% of trivalent chromium. 
     The bioactive compositions of the extracts described above are only representative and may differ depending on the analytical method used, especially so with natural products, as natural products contain multiple bioactives for which reference standards are not readily available in the market. 
     Extraction Methodology 
     In an embodiment, a method of manufacturing of a  T. chebula  fruit extract is described in U.S. Pat. No. 10,500,240, which is incorporated by reference herein for this purpose, to the extent allowed by law. The same method is used for manufacturing  T. bellerica  extract also. 
     In an embodiment, the extraction process of the current invention includes the steps of providing dried fruits of  T. chebula  or  T. bellerica , de-seeding the fruits, pulverizing or grinding the pulp to a powder, extracting the pulp powder with an extraction solvent or solvent mixture, optionally, with heating, to provide a  T. chebula  or  T. bellerica  enriched liquid extract, optionally concentrating the liquid extract and drying the concentrated liquid extract to provide a hydrolyzable tannoid enriched  T. chebula  or  T. bellerica  extract powder. Aqueous solvent is preferred. A particularly preferred solvent is water. Useful extraction temperatures can range from about 25° C. (ambient) to about 90° C. Particularly useful extraction temperatures can range from about 25° C. to about 80° C. 
     In an embodiment, AyuFlex® and Ayuric® are prepared in keeping with the methods described above. 
     In another embodiment, ethanol or methanol or a hydro-alcoholic mixture may be used as the solvent system for extraction. 
     In an embodiment, useful extraction times in conjunction with maintaining useful temperatures can range from about 2 hours to about 16 hours. A particularly useful extraction time range at about 25°±5° C. is from about 12 hours to about 16 hours, and at a temperature of 40°±5° C. is from about 2 hours to about 6 hours. Length and temperature of extraction may be varied at atmospheric pressure (i.e., approx. 1 atm). It is contemplated that pressure can be varied in the extraction process, for example, by use of a commercial pressure reactor apparatus. 
     The extraction process can also include drying the liquid extract to a powder form. Suitable drying methods include spray drying, lyophilization (freeze drying), vacuum drying (with or without heating), evaporation (with or without heating), and concentration under vacuum. Once isolated or obtained, the hydrolyzable tannoid enriched  T. chebula  or  T. bellerica  extract powder may be processed by any suitable means, including grinding, milling, sieving, sizing, blending and the like. The obtained hydrolyzable tannoid enriched  T. chebula  or  T. bellerica  extract powder may be prepared in any suitable particle size or particle size range. 
     Process additives such as microcrystalline cellulose, starch, maltodextrin and the like as carrier materials, anti-adherents such as silicone dioxide, rice bran powder and the like, and preservatives such as sodium benzoate, methyl paraben, propyl paraben, natural preservatives and the like may be added during the extraction process or during the final blending of the dried extract powder. 
     In the case of  Phyllanthus emblica  extract, seeds are removed from the fresh fruits after washing, the juice is separated from the pulp by pressing and/or centrifugation and the juice is dried by spray drying or another suitable drying technique, such as microwave drying, freeze drying, etc. Additives, such as preservatives, such as sodium chloride and sodium benzoate, carrier materials such as maltodextrin, microcrystalline cellulose, anti-adherent such as silicon dioxide and rice bran powder may also be added, optionally. 
     In the cases of  Withania somnifera , Shilajit and  Azadirachta indica , the dried roots and/or leaves of the plant, the shilajit stone and the dried leaves and twigs respectively, are milled and subjected to extraction and drying. Extraction is done using water only as the solvent or an alcohol, preferably ethanol or methanol, followed by water. The liquid extract is then optionally concentrated by evaporation and then dried by spray drying, freeze drying, microwave drying or another suitable drying technique. Extraction temperatures may range from 40° C.±5° C. to 90° C.±5° C., preferably 60° C.±5° C. to 80° C.±5° C. Extraction times may vary from 1 hour to 12 hours, preferably from 2 hours to 6 hours. Several cycles of extraction may also be done. In an embodiment of  Azadirachta indica , only leaves may be used as the starting raw material. 
     Trivalent chromium complex is made by dissolving trivalent chromium chloride in water, with or without heat, adding  Phyllanthus emblica  extract and mixing for a suitable length of time to form a complex, then adding shilajit and mixing for a suitable length of time, then adding a carrier material such as microcrystalline cellulose, maltodextrin, starch, etc. and mixing for a suitable length of time, followed by spray drying, freeze drying, microwave drying or another suitable drying method. The temperature of the complexation step may be from 30° C.±5° C. to 80° C.±5° C., preferably from 40° C.±5° C. to 60° C.±5° C. 
     In an embodiment, a “combination” of this invention refers to a mixture of an extract of the present invention with another extract, and/or a mixture of one or more extract(s) with a known anti-cancer drug, for instance those drugs used as controls and/or identified as standard of care drugs in the Examples. In an embodiment, a combination is in the same composition. In an embodiment, a composition of the present invention includes one or more extracts as described, and the combination of extract with an anti-cancer drug occurs in a subject&#39;s body. The term “combination” is not intended to be limiting in the context of this invention. Combination therapy refers to a regimen for administering an extract including a combination of extracts of this invention with an anti-cancer drug, for instance to improve treatment outcome, and/or to increase anti-cancer effects as compared with the extract(s) alone and the anti-cancer drug alone. In an embodiment, a combination according to the present invention provides a synergistic effect, greater than the effect achieved by its individual components. While references to synergy and synergistic effects may be made throughout this application, all instances of such may not be expressly pointed out. 
     A “dietary supplement” according to the present invention refers to a composition of this invention which is orally administered as an addition to a subject&#39;s diet, which is not a natural or conventional food, which when administered inhibits the proliferation of cancer cells in the subject&#39;s body. In an embodiment, the dietary supplement is administered daily; in an embodiment, the dietary supplement is administered daily for instance for at least 3 days, 5 days, 1 week, 1 day to 1 week, 1 week to 26 weeks, or chronically for at least 1 day to 3 months, 6 months, 9 months, or 1 year or more, or for another period of time according to the present invention. A dietary supplement may be formulated into various forms, as discussed throughout this application. In an embodiment, a dietary supplement of this invention comprises one or more extracts of this invention, such as one or more aqueous standardized extracts or one or more hydroalcoholic extracts of the present invention. 
     EXAMPLES 
     The present invention may be further understood in connection with the below Examples and with embodiments described throughout this application. The following non-limiting Examples and embodiments described below and throughout this application are provided to illustrate the invention. Materials and methods used in Example I were also used with regard to the other Examples, unless indicated or expressly stated otherwise. 
     Example I 
     Anti-proliferation assays were performed on 6 types of primary patient-derived cancer cells (PDCs) from 6 different human donors, as described in Table 1: 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 PDCs from 6 different human donors 
               
            
           
           
               
               
               
               
            
               
                   
                   
                   
                 Donor 
               
               
                   
                   
                 Cancer 
                 Gender/Age 
               
               
                 Cancer Category 
                 PDC 
                 Characteristics 
                 (years) 
               
               
                   
               
               
                 Solid 
                 Glioma 
                 Glioblastoma 
                 M/74 
               
               
                   
                   
                 Grade IV 
                   
               
               
                 Solid 
                 Breast Cancer 
                 ER/PR+ Her2 
                 M/52 
               
               
                   
                   
                 equivocal 
                   
               
               
                   
                   
                 (“HR+”) 
                   
               
               
                 Solid 
                 Breast Cancer 
                 ER/PR− Her2+ 
                 F/45 
               
               
                   
                   
                 (“Her2+”) 
                   
               
               
                 Solid 
                 Breast Cancer 
                 Triple negative 
                 F/42 
               
               
                   
                   
                 (“TN”) 
                   
               
               
                 Hematopoietic 
                 CLL (Chronic 
                 — 
                 M/77 
               
               
                   
                 lymphocytic 
                   
                   
               
               
                   
                 leukemia) 
                   
                   
               
               
                 Hematopoietic 
                 AML (Acute 
                 M4 subtype 
                 F/76 
               
               
                   
                 myeloid 
                   
                   
               
               
                   
                 leukemia) 
               
               
                   
               
            
           
         
       
     
     Tests were performed in 384-well plates (24×16), using 1250 to 10,000 cells/well depending on the size and proliferation rate of different PDCs, in 30 ul of culture media (DMEM/F12 with serum and growth factors), with control wells set aside on each plate for media alone (4 wells), untreated PDCs (4 wells), and for PDCs treated with 10 ul of a known anti-cancer drug as a control. For instance, 10 ul of 40 uM GDC-0941 (pictilisib)), or 40 uM doxorubicin (2 wells each), was used, for a final concentration of 10 uM each well. The outermost rows/columns of the plates were filled with PBS (pH 7.4) to avoid an edge-effect and were considered as blanks. 
     Standardized aqueous extracts identified in Table 2 were prepared in main stock solutions by dissolving 1 mg of extract in 1 ml PBS (phosphate-buffered saline, pH 7.4). 
                     TABLE 2                  Examples of standardized aqueous extracts                         Preparation type   Source   Example               Standardized      Terminalia   chebula  fruits   AyuFlex ®       aqueous extract               Standardized      Terminalia   bellerica  fruits   Ayuric ®       aqueous extract               Standardized      Phyllanthus   emblica  fruits   Capros ®       aqueous extract               Standardized      Withania   somnifera  roots + leaves   Sensoril ®       aqueous extract               Standardized    Shilajit   PrimaVie ®       aqueous extract               Standardized      Azadirachta   indica  leaves + twigs   PhytoBGS ®       aqueous extract                    
All extracts used in this Example were provided by Natreon, Inc. (New Brunswick, N.J.).
 
     Anti-Proliferation Assay Protocol: 30 ul of cells were plated at the density of 1250 to 10,000 cells/well. The plate was incubated in a CO 2  incubator for 0/N incubation for 24 hours. Next day, 10 ul of 40 uM GDC-0941 (pictilisib) or 10 ul of 40 uM doxorubicin were added to incubated cells in the specified control drug wells, and 10 ul of 4× extract solutions were added to incubated cells in 3 concentrations: 400 ug/ml (2 wells), 120 ug/ml (3 wells), 40 ug/ml (3 wells). The final assay concentration of control drug was 10 uM for DGC-0941 (pictilisib) and doxorubicin. Final assay concentrations of made from stock solutions of standardized aqueous extracts identified in Table 2 were 100 ug/ml, 30 ug/ml, and 10 ug/ml. Since 10 ul extract was added to 30 ul of cell suspension, it diluted 4 times, so to maintain the final assay concentrations, 4× concentrated extract solutions (400 ug/ml, 120 ug/ml, and 40 ug/mo) were prepared from main stock (1 mg/ml) and added to the plate. In all experiments, 10 ul of 4 times concentrated stock (4×) was added to wells containing 30 ul of cell suspension to achieve final assay concentrations at  1 X. Other anti-cancer drugs used as controls throughout this Example include temozolomide, docetaxel, 5-fluorouracil (5-FU), ibrutinib, arsenic trioxide, and cytarabine, and were applied to PDCs generally as described. 
     Once extracts were added to incubated cells and mixed gently, the plate was given a brief spin and incubated in the CO 2  incubator for 72 hours. The plate was removed from the incubator and 10 ul of detection reagent (CellTiterGlo®; Promega Corporation, Wisconsin, USA) added to each well of the 384 well plate. The plate was incubated (10 min) and luminescence counts measured, with luminescence produced proportional to the number of viable (living, metabolically active) cells for each assay. The assay described above is a two-dimensional (2D) assay. 
     References to synergy below are not intended as limiting; other instances of synergy may be present in view of the data provided below. 
     Results and Discussion 
     1.  Terminalia chebula  Fruit Extract (AyuFlex®) 
     Table 3 shows the percentage of PDCs inhibited by AyuFlex® (±Standard Deviation) in the anti-proliferation assay described above. The results are pictured in  FIG. 1 . Bars shown from left to right for each cell type represent the application of 100 ug Ayuflex® extract/ml, 30 ug Ayuflex® extract/ml, and 10 ug Ayuflex® extract/ml to the cells. 
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Inhibition of PDCs by AyuFlex ® 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                 Breast 
                 Breast 
                 Breast 
                   
                   
               
               
                 % inhibition ±SD 
                 Glioma 
                 Cancer, HR+ 
                 Cancer, Her2+ 
                 Cancer, TN 
                 CLL 
                 AML 
               
               
                   
               
               
                 100 ug/ml 
                 68 ± 2 
                 64 ± 9 
                 62 ± 8 
                 77 ± 4 
                 95 ± 2 
                 33 ± 0 
               
               
                 30 ug/ml 
                 35 ± 9 
                 50 ± 7 
                 64 ± 7 
                 65 ± 4 
                  52 ± 13 
                  6 ± 8 
               
               
                 10 ug/ml 
                  34 ± 10 
                 43 ± 8 
                 52 ± 3 
                 54 ± 3 
                 26 ± 9 
                 −41 ± 3  
               
               
                   
               
            
           
         
       
     
     Hematopoietic PDCs 
     AyuFlex® showed dose-dependent, high anti-cancer activity against CLL (Chronic lymphocytic leukemia) primary cells, with 50% of CLL cells inhibited by AyuFlex® extract at the 30 ug/ml concentration applied to the CLL cells, and 95% inhibition of CLL proliferation at the 100 ug/ml concentration. AyuFlex® inhibited about 33% of AML (acute myeloid leukemia) cells at the 100 ug/ml concentration. 
     Solid Tumor PDCs 
     AyuFlex® showed potent anti-cancer activity against all solid cancer PDCs. The most potent anti-cancer activity was seen against Breast Cancer PDCs (Triple Negative (“TN”), and ER/PR− Her2+(“Her2+”), with AyuFlex® showing more than 50% inhibition of these cells at the concentration of 10 ug/ml. Anti-cancer activity against the other Breast Cancer PDCs (“HR+”) was also high, showing 50% inhibition at greater than/equal to the 30 ug/ml concentration. Anti-cancer activity against glioma cells were also high, with 35% inhibition at 30 ug/ml and 68% inhibition at the 100 ug/ml concentration. 
     2.  Terminalia bellerica  Fruit Extract (Ayuric®) 
     Table 4 shows the percentage of PDCs inhibited by Ayuric® (±Standard Deviation) in the anti-proliferation assay described above. The results are pictured in  FIG. 2 . Bars shown from left to right for each cell type represent the application of 100 ug Ayuric® extract/ml, 30 ug Ayuric® extract/ml, and 10 ug Ayuric® extract/ml to the cell wells. 
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 Inhibition of PDCs by Ayuric ® 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                 Breast 
                 Breast 
                 Breast 
                   
                   
               
               
                 % inhibition ±SD 
                 Glioma 
                 Cancer, HR+ 
                 Cancer, Her2+ 
                 Cancer, TN 
                 CLL 
                 AML 
               
               
                   
               
               
                 100 ug/ml 
                 47 ± 9 
                 54 ± 11 
                  48 ± 13 
                 65 ± 9 
                 73 ± 8 
                 30 ± 15 
               
               
                 30 ug/ml 
                 27 ± 9 
                 32 ± 9  
                 36 ± 8 
                 47 ± 8 
                  29 ± 10 
                 5 ± 5 
               
               
                 10 ug/ml 
                  20 ± 11 
                 16 ± 13 
                 −1 ± 3 
                  28 ± 10 
                 −3 ± 4 
                 44 ± 11 
               
               
                   
               
            
           
         
       
     
     Hematopoietic Cancer PDCs 
     Ayuric® showed the highest anti-cancer activity against CLL primary cells, with dose-dependent inhibitory activity and 70% inhibition observed with the 100 ug/ml concentration applied to the CLL PDCs. In AML cells, Ayuric® exhibited moderate anti-cancer activity. 
     Solid Tumor PDCs 
     Ayuric® showed good to moderate anti-cancer activity for all solid cancer PDCs. Ayuric® demonstrated dose-response inhibition and about 50% inhibition of breast cancer Triple Negative PDCs. The anti-cancer activity was found to be more moderate against HR+ breast cancer PDCs and glioma cells. 
     3 . Phyllanthus emblica  Fruit Extract (Capros®) 
     Table 5 shows the percentage of PDCs inhibited by Capros® (±Standard Deviation) in the anti-proliferation assay described above. The results are pictured in  FIG. 3 . Bars shown from left to right for each cell type represent the application of 100 ug Capros® extract/ml, 30 ug Capros® extract/ml, and 10 ug Capros® extract/ml to the cell wells. 
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Inhibition of PDCs by Capros ® 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                 Breast 
                 Breast 
                 Breast 
                   
                   
               
               
                 % inhibition ±SD 
                 Glioma 
                 Cancer, HR+ 
                 Cancer, Her2+ 
                 Cancer, TN 
                 CLL 
                 AML 
               
               
                   
               
               
                 100 ug/ml 
                 45 ± 2 
                 60 ± 9 
                 53 ± 6 
                 68 ± 6 
                 75 ± 1 
                  9 ± 0 
               
               
                 30 ug/ml 
                 17 ± 8 
                 34 ± 9 
                 54 ± 7 
                 58 ± 8 
                 15 ± 4 
                 23 ± 6 
               
               
                 10 ug/ml 
                 10 ± 1 
                 30 ± 6 
                 23 ± 5 
                  28 ± 10 
                 −13 ± 1  
                 −5 ± 4 
               
               
                   
               
            
           
         
       
     
     Hematopoietic PDCs 
     Capros® showed dose-response and highest anti-cancer activity against CLL primary cells, showing 75% inhibition at the 100 ug/ml concentration applied to the cells. In AML cells, Capros® inhibited about 25% of AML (acute myeloid leukemia) cells at the 30 ug/ml concentration. 
     Solid Tumor PDCs 
     For all solid cancer PDCs, Capros® showed good to moderate anti-cancer activity. Capros® inhibited Breast Cancer Triple Negative and Her2+ cells by 50% at the 30 ug/ml concentration, and inhibited Glioma and Breast Cancer HR+ cells by 45% and 60% at the 100 ug/ml concentration. 
     4 . Withania somnifera  Roots+Leaves (Sensoril®), Aqueous Extract 
     Table 6 shows the percentage of PDCs inhibited by Sensoril® (±Standard Deviation) in the anti-proliferation assay described above. The results are pictured in  FIG. 4 . Bars shown from left to right for each cell type represent the application of 100 ug Sensoril® extract/ml, 30 ug Sensoril® extract/ml, and 10 ug Sensoril® extract/ml to the cell wells. 
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 Inhibition of PDCs by Sensoril ® 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 % inhibition ± 
                   
                 Breast 
                 Breast 
                 Breast 
                   
                   
               
               
                 SD 
                 Glioma 
                 Cancer HR+ 
                 Cancer, Her2+ 
                 Cancer TN 
                 CLL 
                 AML 
               
               
                   
               
               
                 100 ug/ml 
                 12 ± 15 
                 −2 ± 14 
                 24 ± 0  
                 52 ± 0  
                 14 ± 0  
                 69 ± 2  
               
               
                  30 ug/ml 
                 5 ± 9 
                 −4 ± 12 
                 −9 ± 3  
                  7 ± 11 
                 −18 ± 3  
                 69 ± 13 
               
               
                  10 ug/ml 
                  8 ± 14 
                 −9 ± 18 
                 −9 ± 11 
                 6 ± 8 
                 −3 ± 8  
                 60 ± 13 
               
               
                   
               
            
           
         
       
     
     Hematopoietic PDCs 
     Sensoril® showed very potent activity against AML primary cells, inhibiting ≥50% at 10 ug/ml concentration. Sensoril® did not show much anti-cancer activity for CLL. 
     Solid Tumor PDCs 
     Sensoril® inhibited the proliferation of breast cancer “TN” cells by 50% at the 100 ug/ml concentration. Sensoril® did not show substantial anti-cancer activity for glioma or the other breast cancer cells tested. 
     5. Shilajit (PrimaVie®) 
     Table 7 shows the percentage of PDCs inhibited by PrimaVie® (±Standard Deviation) in the anti-proliferation assay described above. The results are pictured in  FIG. 5 . Bars shown from left to right for each cell type represent the application of 100 ug PrimaVie® extract/ml, 30 ug PrimaVie® extract/ml, and 10 ug PrimaVie® extract/ml to the cell wells. 
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 Inhibition of PDCs by PrimaVie ® 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                 Breast 
                 Breast 
                 Breast 
                   
                   
               
               
                 % inhibition ±SD 
                 Glioma 
                 Cancer, HR+ 
                 Cancer, Her2+ 
                 Cancer, TN 
                 CLL 
                 AML 
               
               
                   
               
               
                 100 ug/ml 
                 17 ± 4 
                 17 ± 6 
                 19 ± 0 
                 20 ± 0  
                 −22 ± 19 
                 56 ± 7 
               
               
                 30 ug/ml 
                  1 ± 13 
                 −22 ± 4  
                 −5 ± 0 
                  0 ± 11 
                 −12 ± 20 
                 64 ± 4 
               
               
                 10 ug/ml 
                 −9 ± 5 
                 21 ± 8 
                 −13 ± 2  
                 1 ± 8 
                  7 ± 17 
                 25 ± 5 
               
               
                   
               
            
           
         
       
     
     Hematopoietic PDCs 
     PrimaVie® showed the highest anti-cancer activity against AML primary cells, inhibiting ≥50% at the 30 ug/ml concentration applied to the PDCs. PrimaVie® showed a lower anti-cancer activity for CLL PDCs. 
     Solid Tumor PDCs 
     PrimaVie® showed a lower anti-cancer activity for all solid tumor PDCs. 
     6 . Azadirachta indica  Leaves+Twigs Extract (PhytoBGS®) 
     Table 8 shows the percentage of PDCs inhibited by Phyto-BGS® (±Standard Deviation) in the anti-proliferation assay described above. The results are pictured in  FIG. 6 . Bars shown from left to right for each cell type represent the application of 100 ug Phyto-BGS® extract/ml, 30 ug Phyto-BGS® extract/ml, and 10 ug Phyto-BGS® extract/ml to the cell wells. 
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 Inhibition of PDCs by Phyto-BGS ® 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                 Breast 
                 Breast 
                 Breast 
                   
                   
               
               
                 % inhibition ±SD 
                 Glioma 
                 Cancer, HR+ 
                 Cancer, Her2+ 
                 Cancer, TN 
                 CLL 
                 AML 
               
               
                   
               
               
                 100 ug/ml 
                 26 ± 4 
                 32 ± 9 
                 34 ± 8 
                 27 ± 18 
                 −8 ± 15 
                 37 ± 5 
               
               
                 30 ug/ml 
                 −1 ± 1 
                  9 ± 16 
                  7 ± 1 
                 13 ± 12 
                 −12 ± 20  
                  54 ± 10 
               
               
                 10 ug/ml 
                  4 ± 12 
                 −13 ± 4  
                 −2 ± 4 
                  9 ± 11 
                 −1 ± 11 
                 28 ± 7 
               
               
                   
               
            
           
         
       
     
     Hematopoietic PDCs 
     PhytoBGS® showed the highest anti-cancer activity against AML primary cells, showing 50% inhibition at the 30 ug/ml concentration applied to the PDCs. PhytoBGS® showed a lower anti-cancer activity for CLL PDCs. 
     Solid Tumor PDCs 
     PhytoBGS® showed a lower anti-cancer activity for all solid tumor PDCs. 
     Further Results: Solid Cancer Cells, Hematopoietic Cancer Cells, Combination Therapies 
       FIGS. 7, 9, 13, 16, 19, and 22  compare the anti-proliferative effect of the 6 different extracts tested on solid cancer cells and hematopoietic cancer cells. Bars shown from left to right for each extract represent the application of 100 ug extract/ml, 30 ug extract/ml, and 10 ug extract/ml to the cell wells. 
       FIG. 7  represents information provided in Tables 9-12B, showing a significant anti-cancer inhibition of proliferation of Glioma (Grade 4 Glioblastoma) cancer cells in the anti-proliferation assay.  FIG. 7  compares the inhibition of proliferation of Glioma PDCs by the 6 different extracts in the assay, showing the highest inhibition by the application of known drugs 10 uM GDC-0941 (pictilisib) and 10 uM doxorubicin. Of the Extracts of the present invention, 100 ug/ml AyuFlex® showed 68% inhibition (±2, SD) of glioma PDC proliferation, as well as 35% (±9 SD) inhibition at the 30 ug/ml concentration and 34% inhibition (±10 SD) at the 10 ug/ml concentration. Capros® and Ayuric® at 100 ug/ml concentrations showed 47 and 45% inhibition of glioma PDC proliferation (±2 and 9, respectively). PhytoBGS®, PrimaVie®, and Sensoril® inhibited glioma PDCs proliferation at the highest concentration (100 ug/ml) at 26%±4, 17%±4, and 12%±15. 10 uM drug controls (GDC-0941 (pictilisib), doxorubicin) are shown at the far right of  FIG. 7 . 
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 Assay Controls 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                   
                 Assay 
                   
               
               
                   
                 Avg 
                 SD 
                 fold 
                 Z′ 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 100% Proliferation (Cell Control) 
                 36996 
                 5004 
                 1298 
                 0.59 
               
               
                 0% Proliferation (Media only) 
                 29 
                 22 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 10 
               
             
            
               
                   
               
               
                 Assay Drug Control 
               
               
                 GDC-0941 @ 10 uM 
               
            
           
           
               
               
            
               
                 Avg 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
               
                 6290 
                 7680 
                 83 
                 79 
                 81 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 11 
               
             
            
               
                   
               
               
                 Assay Drug Control 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
            
               
                 Avg 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
               
                 6872 
                 6345 
                 81 
                 83 
                 82 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 12A 
               
             
            
               
                   
               
               
                 Assay Results 
               
            
           
           
               
               
            
               
                   
                 % Inhibition 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex 
                 66 
                 70 
                 45 
                 27 
                 33 
                 25 
                 44  
                 33 
               
               
                 Capros 
                 43 
                 46 
                 25 
                 9 
                 16 
                 9 
                 33* 
                 11 
               
               
                 Phyto-BGS 
                 24 
                 29 
                  22* 
                 −2 
                 −1 
                 −7 
                 16* 
                 2 
               
               
                 Primavie 
                 15 
                 20 
                  16* 
                 −7 
                 −5 
                 −12 
                 15* 
                 −5 
               
               
                 Sensoril 
                 23 
                  1* 
                 16 
                 0 
                 0 
                 −4 
                 24  
                 6 
               
               
                 Ayuric 
                 53 
                 40 
                 38 
                 21 
                 23 
                 9 
                 32  
                 20 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 12B 
               
             
            
               
                   
               
               
                 Assay Results 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 Avg % 
                   
                 Avg % 
                   
                 Avg % 
                   
               
               
                   
                 Inhibition 
                 SD 
                 Inhibition 
                 SD 
                 Inhibition 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 AyuFlex ® 
                 68 
                 2 
                 35 
                 9 
                 34 
                 10 
               
               
                 Capros ® 
                 45 
                 2 
                 17 
                 8 
                 10 
                 1 
               
               
                 Phyto-BGS ® 
                 26 
                 4 
                 −1 
                 1 
                 −2 
                 6 
               
               
                 PrimaVie ® 
                 17 
                 4 
                 −6 
                 2 
                 −9 
                 5 
               
               
                 Sensoril ® 
                 23 
                 0 
                 5 
                 9 
                 15 
                 12 
               
               
                 Ayuric ® 
                 47 
                 9 
                 27 
                 9 
                 20 
                 11 
               
               
                   
               
            
           
         
       
     
       FIG. 8  represents information provided in Tables 13-16, showing a significant anti-cancer inhibition of glioma cell proliferation. Extracts Ayuric®, Capros®, and Ayuflex® inhibited proliferation of glioma (brain cancer) cells by about 50% or higher, alone, and also in combination with temozolomide, a current and popular drug used to treat glioma. As shown in  FIG. 8 , all three extracts—Ayuric®, Capros®, and Ayuflex®—showed better inhibition than temozolomide when tested alone or in combination. Temozolomide is only effective in about 20% of glioma patients.  FIG. 8  shows that when combined with Ayuric®, Capros®, or Ayuflex®, the effectiveness of temozolomide is approximately doubled. The combination of temozolomide and Ayuric®, Capros®, or Ayuflex® appears to provide a synergistic inhibition of the proliferation of glioma cells. 
     Also,  FIG. 8  shows the first application of Ayuflex® and Capros® combined; Ayuflex® and Ayuric® combined; and Capros® and Ayuric® combined on PDC glioma cells, showing 73% or higher inhibition of proliferation. The combinations of 100 ug/ml Ayuflex® and 100 ug/ml Capros® and of 100 ug/ml Capros® and 100 ug/ml Ayuric® resulted in about 75% and about 73% inhibition of glioma cell proliferation, respectively. The combination of 100 ug/ml Ayuflex® and 100 ug/ml Ayuric® resulted in about 84% inhibition of glioma cell proliferation. Doxorubicin is presented as a control drug at the far right of  FIG. 8 . 
     
       
         
           
               
             
               
                 TABLE 13 
               
               
                   
               
               
                 PLATE MAP 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                 Cell 
                 Ayuflex @ 100 ug/ml 
                 Capros @ 100 ug/ml 
                 Doxorubin @ 10 uM 
               
               
                 Control 
                 Ayuflex @ 100 ug/ml + 
                 Ayuflex @ 100 ug/ml + 
                 Ayuflex @ 100 ug/ml + 
               
               
                   
                 Temozolomide @ 300 uM 
                 Temozolomide @ 100 uM 
                 Temozolomide @ 30 uM 
               
               
                   
                 Capros @ 100 ug/ml + 
                 Capros @ 100 ug/ml + 
                 Capros @ 100 ug/ml + 
               
               
                   
                 Temozolomide @ 300 uM 
                 Temozolomide @ 100 uM 
                 Temozolomide @ 30 uM 
               
               
                   
                 Ayuric @ 100 ug/ml + 
                 Ayuric @ 100 ug/ml + 
                 Ayuric @ 100 ug/ml + 
               
               
                   
                 Temozolomide @ 300 uM 
                 Temozolomide @ 100 uM 
                 Temozolomide @ 30 uM 
               
               
                 Media 
                 Ayuflex @ 100 ug/ml + 
                 Ayuflex @ 100 ug/ml + 
                 Capros @ 100 ug/ml + 
               
               
                   
                 Capros @ 100 ug/ml 
                 Ayuric @ 100 ug/ml 
                 Ayuric @ 100 ug/ml 
               
               
                   
                 Temozolomide @ 300 uM 
                 Temozolomide @ 100 uM 
                 Temozolomide @ 30 uM 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 14 
               
             
            
               
                   
               
               
                 PLATE QUALITY CONTROL 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                   
                 Assay 
                   
               
               
                   
                 Avg 
                 SD 
                 fold 
                 Z′ 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 100% Proliferation (Cell Control)  
                 62598 
                 3313 
                 4317 
                 0.84 
               
               
                 0% Proliferation (Media only) 
                 15 
                 1 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 15 
               
             
            
               
                   
               
               
                 DRUG CONTROL 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                 Avg 
                 % Inhibition 
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 Avg 
               
               
                   
               
               
                 1418 
                 1584 
                 2381 
                 98 
                 97 
                 96 
                 97 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 16 
               
             
            
               
                   
               
               
                 RESULTS OF COMBINATION STUDY 
               
            
           
           
               
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Combination 
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 Avg 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex @ 100 ug/ml 
                 25994 
                 31061 
                 27155 
                 58 
                 50 
                 57 
                 55 
                 4 
               
               
                 Ayuflex @ 100 ug/ml + 
                 23635 
                 28706 
                 24030 
                 62 
                 54 
                 62 
                 59 
                 5 
               
               
                 Temozolomide @ 300 uM 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex @ 100 ug/ml + 
                 19216 
                 26462 
                 26112 
                 69 
                 58 
                 58 
                 62 
                 7 
               
               
                 Temozolomide @ 100 uM 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex @ 100 ug/ml + 
                 20081 
                 25499 
                 23823 
                 68 
                 59 
                 62 
                 63 
                 4 
               
               
                 Temozolomide @ 30 uM 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Capros @ 100 ug/ml 
                 27974 
                 34706 
                 36188 
                 55 
                 45 
                 42 
                 47 
                 7 
               
               
                 Capros @ 100 ug/ml + 
                 30426 
                 34509 
                 30327 
                 51 
                 45 
                 52 
                 49 
                 4 
               
               
                 Temozolomide @ 300 uM 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Capros @ 100 ug/ml + 
                 22936 
                 30624 
                 30757 
                 63 
                 51 
                 51 
                 55 
                 7 
               
               
                 Temozolomide @ 100 uM 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Capros @ 100 ug/ml + 
                 28445 
                 34230 
                 36564 
                 55 
                 45 
                 42 
                 47 
                 7 
               
               
                 Temozolomide @ 30 uM 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuric @ 100 ug/ml + 
                 22662 
                 27655 
                 26418 
                 64 
                 56 
                 58 
                 59 
                 4 
               
               
                 Temozolomide @ 300 uM 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuric @ 100 ug/ml + 
                 21383 
                 26609 
                 26896 
                 66 
                 58 
                 57 
                 60 
                 5 
               
               
                 Temozolomide @ 100 uM 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuric @ 100 ug/ml + 
                 21535 
                 28083 
                 28708 
                 66 
                 55 
                 54 
                 58 
                 6 
               
               
                 Temozolomide @ 30 uM 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex @ 100 ug/ml + 
                 13705 
                 16899 
                 16306 
                 78 
                 73 
                 74 
                 75 
                 3 
               
               
                 Capros @ 100 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex @ 100 ug/ml + 
                 8003 
                 11131 
                 10051 
                 87 
                 82 
                 84 
                 84 
                 3 
               
               
                 Ayuric @ 100 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Capros @ 100 ug/ml + 
                 15730 
                 18388 
                 16458 
                 75 
                 71 
                 74 
                 73 
                 2 
               
               
                 Ayuric @ 100 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Temozolomide @ 300 uM 
                 45178 
                 54144 
                 53028 
                 28 
                 14 
                 15 
                 19 
                 8 
               
               
                 Temozolomide @ 100 uM 
                 36970 
                 45539 
                 44666 
                 41 
                 27 
                 29 
                 32 
                 8 
               
               
                 Temozolomide @ 30 uM 
                 37016 
                 52703 
                 53038 
                 41 
                 16 
                 15 
                 24 
                 15 
               
               
                 Doxorubicin @ 10 uM 
                 1418 
                 1584 
                 2381 
                 98 
                 97 
                 96 
                 97 
                 1 
               
               
                   
               
            
           
         
       
     
       FIG. 9  represents information provided in Tables 17-20B, showing the inhibition of proliferation of breast cancer cells (“HR+”, ER/PR+ Her2 Equivocal) in the anti-proliferation assay.  FIG. 9  compares the inhibition of proliferation of Breast cancer “HR+” PDCs by the 6 different extracts in the assay, showing the highest inhibition by the application of control drugs GDC-0941 (pictilisib, 10 uM) and doxorubicin (10 uM). AyuFlex® showed the most potent anti-proliferative effects on these cancer cells, comparable to GDC-0941 (pictilisib), closely followed by Capros®, and then Ayuric®. Effects by PhytoBGS®, PrimaVie®, and Sensoril® are also depicted in  FIG. 9 . Bars shown from left to right for each extract represent the application of 100 ug extract/ml, 30 ug extract/ml, and 10 ug extract/ml to the cell wells. 
     
       
         
           
               
             
               
                 TABLE 17 
               
             
            
               
                   
               
               
                 Assay Controls 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                   
                 Assay 
                   
               
               
                   
                 Avg 
                 SD 
                 fold 
                 Z′ 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 100% Proliferation (Cell Control) 
                 64398 
                 5624 
                 1966 
                 0.74 
               
               
                 0% Proliferation (Media only) 
                 33 
                 20 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 18 
               
             
            
               
                   
               
               
                 Assay Drug Controls 
               
               
                 GDC-0941 @ 10 uM 
               
            
           
           
               
               
            
               
                 Avg 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
               
                 19603 
                 21420 
                 70 
                 67 
                 68 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 19 
               
             
            
               
                   
               
               
                 Assay Drug Controls 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
            
               
                 Avg 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
               
                 16240 
                 13292 
                 75 
                 79 
                 77 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 20A 
               
             
            
               
                   
               
               
                 Assay Results 
               
            
           
           
               
               
            
               
                   
                 % Inhibition 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex ® 
                 70 
                 57 
                 58 
                 48 
                 45 
                 53 
                 37 
                 39 
               
               
                 Capros ® 
                 67 
                 54 
                 44 
                 30 
                 26 
                 35 
                 33 
                 23 
               
               
                 Phyto-BGS ® 
                 39 
                 26 
                 22 
                 −9 
                 14 
                  8* 
                 −16 
                 −11 
               
               
                 Primavie ® 
                 13 
                 21 
                  6* 
                 −25 
                 −19 
                 26 
                 15 
                 −20 
               
               
                 Sensoril ® 
                 8 
                 −12 
                  3 
                 4 
                 −17 
                 11 
                 −21 
                 −18 
               
               
                 Ayuric ® 
                 61 
                 46 
                 42 
                 29 
                 24 
                 31 
                 12 
                 5 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 20B 
               
             
            
               
                   
               
               
                 Assay Results 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 Avg % 
                   
                 Avg % 
                   
                 Avg % 
                   
               
               
                   
                 inhibition 
                 SD 
                 inhibition 
                 SD 
                 inhibition 
                 SD 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Ayuflex ® 
                 64 
                 9 
                 50 
                 7 
                 43 
                 8 
               
               
                 Capros ® 
                 60 
                 9 
                 34 
                 9 
                 30 
                 6 
               
               
                 Phyto-BGS ® 
                 32 
                 9 
                 9 
                 16 
                 −13 
                 4 
               
               
                 Primavie ® 
                 17 
                 6 
                 −22 
                 4 
                 21 
                 8 
               
               
                 Sensoril ® 
                 −2 
                 14 
                 −4 
                 12 
                 −9 
                 18 
               
               
                 Ayuric ® 
                 54 
                 11 
                 32 
                 9 
                 16 
                 13 
               
               
                   
               
            
           
         
       
     
       FIG. 10  shows a dose-response curve of Ayuflex® on HR+ breast cancer cells (that is, breast cancer cells that are ER/PR+ Her2 equivocal). As shown in  FIG. 9  and Table 20B, AyuFlex® showed 50% inhibition of HR+ breast cancer cells at 30 ug/ml doses, tested at 3 graded doses. The S-shaped dose response curve in  FIG. 10  reveals an IC 50  (Inhibitory Concentration 50) of about 42.41 ug Ayuflex®/ml. Table 21 shows data used to generate the dose-response curve. 
     
       
         
           
               
             
               
                 TABLE 21 
               
             
            
               
                   
               
               
                 Dose-response curve of AyuFlex ® on HR+ breast cancer cells 
               
               
                 Ayuflex DRC 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 Conc. 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 1 
                 200 
                 ug/ml 
                 81 
                 80 
                 100 
                 100 
                 100 
               
               
                 2 
                 100 
                 ug/ml 
                 6192 
                 7135 
                 93 
                 92 
                 93 
               
               
                 3 
                 50 
                 ug/ml 
                 27345 
                 35402 
                 70 
                 61 
                 65 
               
               
                 4 
                 25 
                 ug/ml 
                 51896 
                 68951 
                 42 
                 23 
                 33 
               
               
                 5 
                 12.5 
                 ug/ml 
                 66658 
                 83762 
                 26 
                 7 
                 16 
               
               
                 6 
                 6.3 
                 ug/ml 
                 59876 
                 75427 
                 33 
                 16 
                 25 
               
               
                 7 
                 3.1 
                 ug/ml 
                 72667 
                 96931 
                 19 
                 −8 
                 5 
               
               
                 8 
                 1.6 
                 ug/ml 
                 79455 
                 99926 
                 11 
                 −11 
                 0 
               
               
                   
               
            
           
         
       
     
       FIG. 11  represents information provided in Table 22, showing a significant anti-cancer inhibition of HR+(ER/PR+ Her2equivocal) proliferation in combination with docetaxel (an antimicrotubule agent that inhibits spindle assembly during mitosis) or 5-fluorouracil (5-FU). 
     
       
         
           
               
             
               
                 TABLE 22 
               
             
            
               
                   
               
               
                 Inhibition of HR+ with AyuFlex ®, Capros ®, Ayuric ® 
               
            
           
           
               
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Combination 
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 Avg 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex @ 30 ug/ml + 
                 24851 
                 31307 
                 24077 
                 72 
                 65 
                 73 
                 70 
                 4 
               
               
                 Docetaxel @ 3 uM 
               
               
                 Capros @ 100 ug/ml + 
                 18138 
                 25149 
                 23083 
                 80 
                 72 
                 74 
                 75 
                 4 
               
               
                 Docetaxel @ 3 uM 
               
               
                 Ayuric @ 100 ug/ml + 
                 12153 
                 16841 
                 17627 
                 86 
                 81 
                 80 
                 83 
                 3 
               
               
                 Docetaxel @ 3 uM 
               
               
                 Docetaxel @ 3 uM 
                 29734 
                 41946 
                 37833 
                 67 
                 53 
                 58 
                 59 
                 7 
               
               
                 Ayuflex @ 30 ug/ml + 
                 39229 
                 51560 
                 50871 
                 56 
                 43 
                 43 
                 47 
                 8 
               
               
                 5-FU @ 3 uM 
               
               
                 Capros @ 100 ug/ml + 
                 29313 
                 39767 
                 37433 
                 67 
                 56 
                 58 
                 60 
                 6 
               
               
                 5-FU @ 3 uM 
               
               
                 Ayuric @ 100 ug/ml + 
                 19508 
                 30418 
                 27860 
                 78 
                 66 
                 69 
                 71 
                 6 
               
               
                 5-FU @ 3 uM 
               
               
                 5-FU @ 3 uM 
                 51975 
                 71897 
                 69012 
                 42 
                 20 
                 23 
                 28 
                 12 
               
               
                 Ayuflex @ 30 ug/ml + 
                 17047 
                 23164 
                 19543 
                 81 
                 74 
                 78 
                 78 
                 3 
               
               
                 Capros @ 100 ug/ml 
               
               
                 Ayuflex @ 30 ug/ml + 
                 7683 
                 14692 
                 14489 
                 91 
                 84 
                 84 
                 86 
                 4 
               
               
                 Ayuric @ 100 ug/ml 
               
               
                 Capros @ 100 ug/ml + 
                 9840 
                 13176 
                 12303 
                 89 
                 85 
                 86 
                 87 
                 2 
               
               
                 Ayuric @ 100 ug/ml 
               
               
                 Doxorubicin @ 10 uM 
                 17135 
                 18734 
                 22648 
                 81 
                 79 
                 75 
                 78 
                 3 
               
               
                   
               
            
           
         
       
     
       FIG. 11  shows that all three extracts—Ayuflex®, Capros®, and Ayuric®—in combination with docetaxel, showed increased, synergistic inhibition of HR+ breast cancer cell proliferation than when tested alone. 3 uM docetaxel applied to HR+ breast cancer cells in the antiproliferation assay described above inhibited HR+ breast cancer cells by about 59%. As shown in  FIG. 9  and Table 20B above, Ayuflex® alone (30 ug/ml), Capros® alone (100 ug/ml), and Ayuric® alone (100 ug/ml) inhibited HR+breast cancer cells by about 50%, 60%, and 54%, respectively.  FIG. 11  shows that extracts Ayuflex® (30 ug/ml), Capros® (100 ug/ml), and Ayuric® (100 ug/ml), combined with 3 uM docetaxel, inhibited proliferation of HR+ breast cancer cells by about 70%, 75%, and 83% respectively. 
       FIG. 11  also shows that all three extracts—Ayuflex®, Capros®, and Ayuric®—in combination with 5-fluorouracil (5-FU), showed increased, synergistic inhibition of HR+ breast cancer cell proliferation than when tested alone. 3 uM 5-FU applied to HR+ breast cancer cells in the antiproliferation assay described above inhibited HR+ breast cancer cells by about 28%. As shown in  FIG. 9  and Table 20B above, Ayuflex® alone (30 ug/ml), Capros® alone (100 ug/ml), and Ayuric® alone (100 ug/ml) inhibited HR+ breast cancer cells by about 50%, 60%, and 54%, respectively.  FIG. 11  shows that extracts Ayuflex® (30 ug/ml), Capros® (100 ug/ml), and Ayuric® (100 ug/ml), combined with 3 uM 5-FU, inhibited proliferation of HR+ breast cancer cells by about 47%, 60%, and 71% respectively. 
     Also,  FIG. 11  shows that Ayuflex® and Capros® combined; Ayuflex® and Ayuric® combined; and Capros® and Ayuric® combined substantially increased inhibition of HR+ breast cancer cell proliferation over Ayuflex® (50%), Capros® (60%), and Ayuric® (54%) alone. The combination of 30 ug/ml Ayuflex® and 100 ug/ml Capros® resulted in about 78% inhibition of HR+ breast cancer cell proliferation, while the combination of 30 ug/ml Ayuflex® and 100 ug/ml Ayuric®, and of 100 ug/ml Capros® and 100 ug/ml Ayuric® inhibited HR+ breast cancer cell inhibition by about 86% and 87%, respectively. Control drug doxorubicin (10 uM) is shown as inhibiting HR+ breast cancer cell proliferation by 78%, at the far right of  FIG. 11 . 
       FIG. 12  compiles results shown in  FIG. 11  and Table 22 according to the extract studied (that is, Ayuflex®, Capros®, Ayuric®). Ayuflex® (30 ug/ml) combined with 3 uM docetaxel inhibited proliferation of HR+ breast cancer cells by 70%, compared with 59% inhibition by docetaxel alone, 50% inhibition by Ayuflex® alone, and 47% inhibition when combined with 3 uM 5-FU. 3 uM 5-FU alone inhibited cell proliferation by 29%. 
     Capros® (100 ug/ml) combined with 3 uM docetaxel inhibited proliferation of HR+ breast cancer cells by 75%, compared with 59% inhibition by docetaxel alone, 60% inhibition by Capros® alone, and 60% inhibition when combined with 3 uM 5-FU. 3 uM 5-FU alone inhibited cell proliferation by 29%. 
     Ayuric® (100 ug/ml) combined with 3 uM docetaxel inhibited proliferation of HR+ breast cancer cells by 83%, compared with 59% inhibition by docetaxel alone, 54% inhibition by Ayuric® alone, and 71% inhibition when combined with 3 uM 5-FU. 3 uM 5-FU alone inhibited cell proliferation by 29%. 
     Each of the 3 extracts studied—Ayuflex®, Capros®, Ayuric®—showed better inhibition when used in combination with marketed drugs docetaxel and 5-FU than compared to the marketed drugs alone. Also, when these extracts were tested in the antiproliferative assay in combination with each other, for any combination of 2 of the extracts (Ayuflex® 30 ug/ml, Capros® 100 ug/ml, Ayuric® 100 ug/ml), a very good inhibition of HR+ breast cancer cell proliferation was observed, at greater than 78%. See  FIG. 12 , showing the combination of Ayuflex® and Capros® as inhibiting cell proliferation by 78%, Ayuflex® and Ayuric® at 86%, and Capros® and Ayuric® at 87%. For comparison, 10 uM doxorubicin is shown at the far right of  FIG. 12  as inhibiting proliferation of these cells by 78%. 
     As shown in  FIG. 9 , when tested at 3 different doses, Ayuflex® showed the highest anti-cancer activity for HR+ breast cancer cells, followed by Capros® and then Ayuric®. Accordingly, the order of anti-cancer activity for the extracts alone is Ayuflex®&gt;Capros®&gt;Ayuric®. However, in  FIGS. 11 and 12 , when these extracts were tested on the same HR+ breast cancer PDCs in combination with docetaxel and 5-FU, surprisingly, Ayuric® showed the best additive activity, followed by Capros®, and then Ayuflex®. Accordingly, the order of anti-cancer activity for the combination of these extracts with docetaxel or 5-FU is Ayuric®+Drug X&gt;Capros®+Drug X&gt;Ayuflex®+Drug X. 
     Overall, all the three extracts—Ayuflex®, Capros®, and Ayuric®—showed strong anti-cancer activity against HR+ breast cancer PDCs. The cells are highly aggressive in culture; the finding and confirmation that each extract can substantially inhibit the PDCs proliferation in significant percentages in 72 hours, alone or in combination, was surprising and unexpected. 
       FIG. 13  represents information provided in Tables 23-26B, showing the inhibition of proliferation of breast cancer cells (“Her2+”, ER/PR− Her2+) in the anti-proliferation assay described above.  FIG. 13  compares the inhibition of proliferation of Breast cancer “Her2+” PDCs by the 6 different extracts in the assay. The result of application of 10 uM GDC-0941 (pictilisib) and 10 uM doxorubicin as anti-cancer drug controls are shown to the right of the Figure. AyuFlex® showed the most potent anti-proliferative effects of the 6 extracts tested on these cancer cells, comparable to doxorubicin, closely followed by Capros®, and then Ayuric®. Effects by PhytoBGS®, PrimaVie®, and Sensoril® are also depicted in  FIG. 13 . 
     
       
         
           
               
             
               
                 TABLE 23 
               
             
            
               
                   
               
               
                 Assay Controls 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                   
                 Assay  
                   
               
               
                   
                 Avg 
                 SD 
                 fold 
                 Z′ 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 100% Proliferation (Cell Control) 
                 75052 
                 7174 
                 1464 
                 0.71 
               
               
                 0% Proliferation (Media only) 
                 51 
                 22 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 24 
               
             
            
               
                   
               
               
                 Assay Drug Control 
               
               
                 GDC-0941 @ 10 uM 
               
            
           
           
               
               
            
               
                 Avg 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
               
                 21823 
                 23075 
                 71 
                 69 
                 70 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 25 
               
             
            
               
                   
               
               
                 Assay Drug Control 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
            
               
                 Avg 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
               
                 28714 
                 31853 
                 62 
                 58 
                 60 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 26A 
               
             
            
               
                   
               
               
                 Assay Results 
               
            
           
           
               
               
            
               
                   
                 % Inhibition 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex ® 
                 67 
                 56 
                 72  
                 60 
                 60 
                 53  
                 49 
                 54 
               
               
                 Capros ® 
                 57 
                 48 
                 62  
                 51 
                 49 
                 26  
                 18 
                 24 
               
               
                 Phyto-BGS ® 
                 29 
                 40 
                 35* 
                 8 
                 7 
                 3 
                 −5 
                 −4 
               
               
                 Primavie ® 
                 19 
                 −1 
                 11* 
                 −5 
                 −5 
                 13* 
                 −12 
                 −14 
               
               
                 Sensoril ® 
                 24 
                 −6 
                 11* 
                 −12 
                 −7 
                 3 
                 −14 
                 −17 
               
               
                 Ayuric ® 
                 57 
                 39 
                 44  
                 28 
                 37 
                 13* 
                 −3 
                 2 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 26B 
               
             
            
               
                   
               
               
                 Assay Results 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 Avg % 
                   
                 Avg % 
                   
                 Avg % 
                   
               
               
                   
                 inhibition 
                 SD 
                 inhibition 
                 SD 
                 inhibition 
                 SD 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Ayuflex ® 
                 62 
                 8 
                 64 
                 7 
                 52 
                 3 
               
               
                 Capros ® 
                 53 
                 6 
                 54 
                 7 
                 23 
                 5 
               
               
                 Phyto-BGS ® 
                 34 
                 8 
                 7 
                 1 
                 −2 
                 4 
               
               
                 Primavie ® 
                 19 
                 0 
                 −5 
                 0 
                 −13 
                 2 
               
               
                 Sensoril ® 
                 24 
                 0 
                 −9 
                 3 
                 −9 
                 11 
               
               
                 Ayuric ® 
                 48 
                 13 
                 36 
                 8 
                 −1 
                 3 
               
               
                   
               
            
           
         
       
     
     As Ayuflex® showed the highest anti-cancer, anti-proliferative activity of the extracts tested in  FIG. 13 , testing with the aim of revealing a dose-response curve and the IC 50  of Ayuflex® for Her2+ breast cancer cells was performed.  FIG. 14  shows a dose-response curve of Ayuflex® on Her2+ breast cancer cells. As shown in  FIG. 13 , AyuFlex® showed 50% inhibition of TN breast cancer cells at a 10 ug/ml dose, and was tested at 3 graded doses. The dose response curve in  FIG. 14  reveals an IC 50  of about 68.27 ug Ayuflex®/ml. Table 27 shows data used to generate the dose-response curve. 
     
       
         
           
               
             
               
                 TABLE 27 
               
             
            
               
                   
               
               
                 Ayuflex Dose-Response Curve 
               
               
                 Ayuflex DRC 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 Conc. 
                   
                 Raw data 
                 % Inhibition 
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 (ug/ml) 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 1 
                 200 
                 4617 
                 8293 
                 86 
                 75 
                 81 
               
               
                 2 
                 67 
                 17613 
                 20848 
                 47 
                 37 
                 42 
               
               
                 3 
                 22 
                 27891 
                 32137 
                 16 
                 4 
                 10 
               
               
                 4 
                 7 
                 28432 
                 30269 
                 15 
                 9 
                 12 
               
               
                 5 
                 2 
                 25593 
                 27659 
                 23 
                 17 
                 20 
               
               
                 6 
                 1 
                 34443 
                 32209 
                 −3 
                 3 
                 0 
               
               
                 7 
                 0 
                 30800 
                 33752 
                 8 
                 −1 
                 3 
               
               
                 8 
                 0 
                 27215 
                 30136 
                 18 
                 10 
                 14 
               
               
                   
               
            
           
         
       
     
       FIG. 15  represents information provided in Table 28, showing a significant anti-cancer inhibition of Her2+ breast cancer cell proliferation by extracts in combination with 3 uM docetaxel or 3 uM 5-fluorouracil (5-FU). Ayuflex® and docetaxel together inhibited Her2+ cell proliferation by 66% and Ayuric® and docetaxel by 68%, with Capros® and docetaxel inhibiting Her2+ by about 61%. As shown in  FIG. 15 , docetaxel alone inhibited proliferation of the Her2+ cells by about 50%. 
     In contrast, 5-FU in combination with Ayuflex® (30 ug/ml) and Capros® (100 ug/ml), inhibited proliferation by about 24% and 27% respectively, as compared with 18% by 5-FU alone.  FIG. 13  shows that Ayuric® inhibited proliferation of Her2+ cells by about 34%, however, Ayuric® and 5-FU together appeared to work synergistically, inhibiting proliferation by about 56%. 
     Finally, combinations of the three extracts share similarities in their inhibition of Her2+ cells (57%, 47%, 46% respectively for Ayuflex®/Capros®, Ayuflex®/Ayuric®, and Capros®/Ayuric®). With the exception of the combination of Ayuric® and 5-FU, the extracts share similarities in their inhibition of Her2+ cells when used in combination with docetaxel and 5-FU. It is noted that the PDCs used in this experiment were slow-growing and differed in morphology. Also, a new stock of Ayuflex® was prepared for this study, which showed a little less potency than stock used in some previous experiments. 
     
       
         
           
               
             
               
                 TABLE 28 
               
             
            
               
                   
               
               
                 Inhibition of Her2+ breast cancer cell proliferation 
               
            
           
           
               
               
               
               
            
               
                   
                 Raw data 
                 % inhibition 
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Combination 
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 Avg 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex @30 ug/ml + 
                 8318 
                 13508 
                 11954 
                 75 
                 60 
                 64 
                 66 
                 8 
               
               
                 Docetaxel @ 3 uM 
               
               
                 Capros @100 ug/ml + 
                 11793 
                 13674 
                 13069 
                 65 
                 59 
                 61 
                 61 
                 3 
               
               
                 Docetaxel @ 3 uM 
               
               
                 Ayuric @ 100 ug/ml + 
                 9029 
                 12954 
                 10392 
                 73 
                 61 
                 69 
                 68 
                 6 
               
               
                 Docetaxel @ 3 uM 
               
               
                 Docetaxel @ 3 uM 
                 23284 
                 18884 
                 14463 
                  30* 
                 43 
                 57 
                 50 
                 9 
               
               
                 Ayuflex @ 30 ug/ml + 
                 24373 
                 26114 
                 34166 
                 27 
                 22 
                 −3 
                 24 
                 4 
               
               
                 5-FU@ 3 uM 
               
               
                 Capros @ 100 ug/ml + 
                 22889 
                 25557 
                 17709 
                 31 
                 23 
                  47* 
                 27 
                 6 
               
               
                 5-FU @ 3 uM 
               
               
                 Ayuric @ 100 ug/ml + 
                 15454 
                 20125 
                 14086 
                 54 
                  40* 
                 58 
                 56 
                 3 
               
               
                 5-FU @ 3 uM 
               
               
                 5-FU @ 3 uM 
                 19643 
                 28153 
                 26851 
                  41* 
                 16 
                 19 
                 18 
                 3 
               
               
                 Ayuflex @ 30 ug/ml + 
                 13209 
                 19318 
                 15708 
                 60 
                  42* 
                 53 
                 56 
                 5 
               
               
                 Capros @ 100 ug/ml 
               
               
                 Ayuflex @ 30 ug/ml + 
                 12903 
                 17628 
                 17705 
                  61* 
                 47 
                 47 
                 47 
                 0 
               
               
                 Ayuric @ 100 ug/ml 
               
               
                 Capros @ 100 ug/ml + 
                 11840 
                 17003 
                 18991 
                  65* 
                 49 
                 43 
                 46 
                 4 
               
               
                 Ayuric @ 100 ug/ml 
               
               
                 Doxorubicin @ 10 uM 
                 1530 
                 1667 
                 996 
                 95 
                 95 
                 97 
                 96 
                 1 
               
               
                   
               
               
                 *Outliers 
               
            
           
         
       
     
       FIG. 16  represents information provided in Tables 29-32B, showing the inhibition of proliferation of breast cancer cells (“TN”, Triple Negative) in the anti-proliferation assay.  FIG. 16  compares the inhibition of proliferation of Breast cancer “Triple Negative” PDCs by the 6 different extracts in the assay, with GDC-0941 (pictilisib) and doxorubicin shown at the right side of  FIG. 16  as drug controls known to inhibit the proliferation of cancer cells. AyuFlex® showed the most potent anti-proliferative effects on these cancer cells, comparable to GDC-0941 and doxorubicin, closely followed by Capros®, Ayuric®, and then Sensoril®, showing anti-proliferative activity in the 100 ug/ml concentration applied to cells. Effects by PhytoBGS® and PrimaVie® are also depicted in  FIG. 16 . 
     
       
         
           
               
             
               
                 TABLE 29 
               
             
            
               
                   
               
               
                 Assay Controls 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                   
                 Assay 
                   
               
               
                   
                 Avg 
                 SD 
                 fold 
                 Z′ 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 100% Proliferation (Cell Control) 
                 71278 
                 6637 
                 1262 
                 0.72 
               
               
                 0% Proliferation (Media only) 
                 57 
                 18 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 30 
               
             
            
               
                   
               
               
                 Assay Drug Control 
               
               
                 GDC-0941 @ 10 uM 
               
            
           
           
               
               
            
               
                 Avg 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
               
                 11517 
                 12897 
                 84 
                 82 
                 83 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 31 
               
             
            
               
                   
               
               
                 Assay Drug Control 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
            
               
                 Avg 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
               
                 19364 
                 19643 
                 73 
                 72 
                 73 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 32A 
               
             
            
               
                   
               
               
                 Assay Results 
               
            
           
           
               
               
            
               
                   
                 % Inhibition 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex ® 
                 79 
                 74 
                 70 
                 63 
                 63 
                 58 
                 52 
                 53 
               
               
                 Capros ® 
                 72 
                 64 
                 66 
                 55 
                 52 
                 39 
                 22 
                 24 
               
               
                 Phyto-BGS ® 
                 40 
                 15 
                 26 
                 2 
                 10 
                 22 
                 3 
                 3 
               
               
                 Primavie ® 
                 84 
                 20 
                 9 
                 −12 
                 2 
                 10 
                 −4 
                 −3 
               
               
                 Sensoril ® 
                 52 
                 19 
                 19 
                 −3 
                 4 
                 16 
                 0 
                 2 
               
               
                 Ayuric ® 
                 71 
                 58 
                 56 
                 41 
                 45 
                 38 
                 18 
                 27 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 32B 
               
             
            
               
                   
               
               
                 Assay Results 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 Avg % 
                   
                 Avg % 
                   
                 Avg % 
                   
               
               
                   
                 inhibition 
                 SD 
                 inhibition 
                 SD 
                 inhibition 
                 SD 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Ayuflex ® 
                 77 
                 4 
                 65 
                 4 
                 54 
                 3 
               
               
                 Capros ® 
                 68 
                 6 
                 58 
                 8 
                 28 
                 10 
               
               
                 Phyto-BGS ® 
                 27 
                 18 
                 13 
                 12 
                 9 
                 11 
               
               
                 Primavie ® 
                 20 
                 0 
                 0 
                 11 
                 1 
                 8 
               
               
                 Sensoril ® 
                 35 
                 24 
                 7 
                 11 
                 6 
                 8 
               
               
                 Ayuric ® 
                 65 
                 9 
                 47 
                 8 
                 28 
                 10 
               
               
                   
               
            
           
         
       
     
     As Ayuflex® showed the highest anti-cancer, anti-proliferative activity of the extracts tested in  FIG. 16 , testing with the aim of revealing a dose-response curve and the IC 50  of Ayuflex® for TN breast cancer cells was performed.  FIG. 17  shows a dose-response curve of Ayuflex® on TN (Triple Negative) breast cancer cells. AyuFlex® showed 50% inhibition of TN breast cancer cells at a 10 ug/ml dose, when tested at 3 graded doses. The dose response curve in  FIG. 17  reveals an IC 50  of about 54.61 ug Ayuflex®/ml. Table 33 shows data used to generate the dose-response curve. 
     
       
         
           
               
             
               
                 TABLE 33 
               
             
            
               
                   
               
               
                 Ayuflex ® Dose-Response Curve 
               
               
                 Ayuflex DRC 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 Conc. 
                   
                 Raw data 
                 % Inhibition 
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 (ug/ml) 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 1 
                 200 
                 12870 
                 15012 
                 83 
                 80 
                 82 
               
               
                 2 
                 66.7| 
                 34601 
                 36098 
                 54 
                 52 
                 53 
               
               
                 3 
                 22.2 
                 51275 
                 55608 
                 32 
                 26 
                 29 
               
               
                 4 
                 7.4 
                 55229 
                 62697 
                 27 
                 17 
                 22 
               
               
                 5 
                 2.5 
                 65914 
                 76238 
                 13 
                 −1 
                 6 
               
               
                 6 
                 0.8 
                 67950 
                 75290 
                 10 
                 0 
                 5 
               
               
                 7 
                 0.3 
                 74107 
                 80541 
                 2 
                 −7 
                 −2 
               
               
                 8 
                 0.1 
                 67946 
                 74190 
                 10 
                 2 
                 6 
               
               
                   
               
            
           
         
       
     
       FIG. 18  represents information provided in Table 34, showing a significant anti-cancer inhibition of TN breast cancer cell proliferation in combination with 3 uM docetaxel or 3 uM 5-fluorouracil (5-FU). Ayuflex® (10 ug/ml) and docetaxel together inhibited TN cell proliferation by 63% and Ayuric® (30 ug/ml) and docetaxel by 67%, with Capros® (30 ug/ml) and docetaxel inhibiting TN cell proliferation by about 57%. Docetaxel alone inhibited proliferation of the TN cells by about 48%. In a previous study, Ayuflex® (10 ug/ml) inhibited TN cell proliferation by 54%; Capros® inhibited TN cell proliferation by 58%; and Ayuric® inhibited TN cell proliferation by 47%. The inhibition of proliferation by docetaxel and Ayuric® in particular shows synergistic results, as alone, each substance inhibited TN cell proliferation by 47-48%, but taken together, inhibition of TN cell proliferation increased nearly 20% to 67%. 
       FIG. 18  also shows that 5-FU (3 uM) in combination with Ayuflex® (10 ug/ml) or with Ayuric® (30 ug/ml), inhibited TN cell proliferation by 54%; and in combination with Capros®, inhibited TN cell proliferation by 49%, as compared with 38% by 5-FU alone. In addition,  FIG. 18  shows that combinations of the three extracts inhibited TN cell proliferation by 55%, 51%, 55% respectively for Ayuflex® (10 ug/ml)/Capros® (30 ug/ml), Ayuflex® (10 ug/ml)/Ayuric® (30 ug/ml), and Capros® (30 ug/ml)/Ayuric® (30 ug/ml). Overall, the 3 extracts share similarities in their inhibition of TN cells, when tested alone, in combination with docetaxel and 5-FU, or in combination with each other. 
     
       
         
           
               
             
               
                 TABLE 34 
               
             
            
               
                   
               
               
                 Inhibition of TN breast cancer cell proliferation 
               
            
           
           
               
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Combination 
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 Avg 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex @ 10 ug/ml + 
                 24205 
                 29046 
                 29650 
                 68 
                 62 
                 61 
                 63 
                 4 
               
               
                 Docetaxel @ 3 uM 
               
               
                 Capros @ 30 ug/ml + 
                 26716 
                 35847 
                 35282 
                 65 
                 53 
                 53 
                 57 
                 7 
               
               
                 Docetaxel @ 3 uM 
               
               
                 Ayuric @ 30 ug/ml + 
                 19960 
                 26958 
                 27891 
                 74 
                 64 
                 63 
                 67 
                 6 
               
               
                 Docetaxel @ 3 uM 
               
               
                 Docetaxel @ 3 uM 
                 33410 
                 42476 
                 42703 
                 56 
                 44 
                 43 
                 48 
                 7 
               
               
                 Ayuflex @ 10 ug/ml + 
                 29616 
                 38064 
                 36091 
                 61 
                 50 
                 52 
                 54 
                 6 
               
               
                 5-FU @ 3 uM 
               
               
                 Capros @ 30 ug/ml + 
                 32709 
                 43186 
                 40709 
                 57 
                 43 
                 46 
                 49 
                 7 
               
               
                 5-FU @ 3 uM 
               
               
                 Ayuric @ 30 ug/ml + 
                 28675 
                 36109 
                 38462 
                 62 
                 52 
                 49 
                 54 
                 7 
               
               
                 5-FU @ 3 uM 
               
               
                 5-FU @ 3 uM 
                 39077 
                 50221 
                 50264 
                 48 
                 33 
                 33 
                 38 
                 9 
               
               
                 Ayuflex @ 10 ug/ml + 
                 29096 
                 36274 
                 36747 
                 61 
                 52 
                 51 
                 55 
                 6 
               
               
                 Capros @ 30 ug/ml 
               
               
                 Ayuflex @ 10 ug/ml + 
                 32247 
                 41305 
                 37502 
                 57 
                 45 
                 50 
                 51 
                 6 
               
               
                 Ayuric @ 30 ug/ml 
               
               
                 Capros @ 30 ug/ml + 
                 27434 
                 38949 
                 35603 
                 64 
                 48 
                 53 
                 55 
                 8 
               
               
                 Ayuric @ 30 ug/ml 
               
               
                 Doxorubicin @ 10 uM 
                 4420 
                 6305 
                 5874 
                 91 
                 92 
                 92 
                 93 
                 1 
               
               
                   
               
            
           
         
       
     
       FIG. 19  represents information provided in Tables 35-38B, showing the inhibition of proliferation of chronic lymphocytic leukemia (CLL) PDCs in the anti-proliferation assay.  FIG. 19  compares the inhibition of proliferation of CLL PDCs by the 6 different extracts in the anti-proliferation assay, showing nearly 100% inhibition of AML cell proliferation with the 100 ug/ml AyuFlex® extract concentration, and anti-proliferative activity of AyuFlex® comparable to drug standards GDC-0941 (pictilisib) and doxorubicin at the 30 ug/ml concentration. Capros® and Ayuric® outperformed the drug standards at their 100 ug/ml concentrations, showing about 75% anti-proliferation activity. Effects by PhytoBGS®, PrimaVie®, and Sensoril® are also depicted in  FIG. 19 . 
     
       
         
           
               
             
               
                 TABLE 35 
               
             
            
               
                   
               
               
                 Assay Controls 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 Assay 
                   
               
               
                   
                   
                 Avg 
                 SD 
                 fold 
                 Z′ 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 100% Proliferation (Cell Control) 
                 5930 
                 532 
                 84 
                 0.71 
               
               
                   
                 0% Proliferation (Media only) 
                 70 
                 28 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 36 
               
             
            
               
                   
               
               
                 Assay Drug Control 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
            
               
                 Avg 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
               
                 3002 
                 2367 
                 50 
                 61 
                 55 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 37 
               
             
            
               
                   
               
               
                 Assay Drug Control 
               
               
                 GDC-0941 @ 10 uM 
               
            
           
           
               
               
            
               
                 Avg 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
               
                 2562 
                 3477 
                 57 
                 42 
                 50 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 38A 
               
             
            
               
                   
               
               
                 Assay Results 
               
            
           
           
               
               
            
               
                   
                 % Inhibition 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex ® 
                 97 
                 94 
                 66  
                 41 
                 49 
                 36  
                 24 
                 18 
               
               
                 Capros ® 
                 76 
                 74 
                 38* 
                 12 
                 19 
                 19* 
                 −13 
                 −12 
               
               
                 Phyto-BGS ® 
                 2 
                 −19 
                 10  
                 −20 
                 −27 
                 12  
                 −6 
                 −8 
               
               
                 Primavie ® 
                 −9 
                 −36 
                 9 
                 −12 
                 −31 
                 26  
                 −3 
                 −3 
               
               
                 Sensoril ® 
                 14 
                 −16 
                 4 
                 −16 
                 −20 
                 3 
                 −43 
                 −8 
               
               
                 Ayuric ® 
                 78 
                 67 
                 39  
                 25 
                 21 
                 18* 
                 −6 
                 0 
               
               
                   
               
            
           
         
       
     
                     TABLE 38B                  Assay Results                                 100 ug/ml   30 ug/ml   10 ug/ml                                             Avg %       Avg %       Avg %               inhibition   SD   inhibition   SD   inhibition   SD                                                     Ayuflex   95   2   52   13   26   9       Capros   75   1   15   4   −13   1       Phyto-BGS   −8   15   −12   20   −1   11       Primavie   −22   19   −12   20   7   17       Sensoril   −1   21   −11   13   −16   24       Ayuric   73   8   29   10   −3   4                    
As previous testing showed Ayuflex®, Capros®, and Ayuric® shared good activity for inhibiting CLL PDC proliferation in the anti-proliferative assay discussed above, testing with the aim of revealing a dose-response curve and the IC 50  of Ayuflex®, Capros®, and Ayuric® for CLL PDCs was performed.  FIG. 20  shows a dose-response curve of Ayuflex® on CLL cells, and an IC 50  of 21.95 ug/ml. The 22 ug/ml IC 50  for Ayuflex® was in line with about 50% inhibition observed with Ayuflex® previously. Also, an IC 50  of 80.82 ug/ml was identified for Capros® and for Ayuric®, an IC 50  of about 93.46 ug/ml. Tables 40-42 show data used to generate the dose-response curves for the extracts. A dose-response curve of the inhibition of CLL cells by ibrutinib is also shown in  FIG. 20 , and data to generate the curve shown in Table 39.
 
     
       
         
           
               
             
               
                 TABLE 39 
               
             
            
               
                   
               
               
                 Dose-response curve 
               
               
                 Ibrutinib DRC 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 Conc. 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 1 
                 6.00E−05 
                 20364 
                 23884 
                 49 
                 40 
                 44 
               
               
                 2 
                 3.00E−05 
                 31715 
                 32913 
                 20 
                 17 
                 18 
               
               
                 3 
                 1.50E−05 
                 33454 
                 37044 
                 15 
                  6 
                 11 
               
               
                 4 
                 7.50E−06 
                 36853 
                 40636 
                  7 
                 −3 
                 2 
               
               
                 5 
                 3.75E−06 
                 41353 
                 41370 
                 −5 
                 −5 
                 −5 
               
               
                 6 
                 1.88E−06 
                 43486 
                 44692 
                 −10  
                 −13  
                 −12 
               
               
                 7 
                 9.38E−07 
                 157228 
                 77748 
                 −299*  
                 −97* 
               
               
                 8 
                 4.69E−07 
                 78043 
                 49566 
                 −98* 
                 −26  
                 −26 
               
               
                   
               
               
                 *Outliers 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 40 
               
             
            
               
                   
               
               
                 Dose-response curve 
               
               
                 Ayuflex DRC 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 Conc. 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 1 
                 200 
                 ug/ml 
                 1274 
                 1720 
                 97 
                 96 
                 96 
               
               
                 2 
                 100 
                 ug/ml 
                 5815 
                 132833 
                 85 
                 −237*  
                 85 
               
               
                 3 
                 50 
                 ug/ml 
                 9035 
                 45631 
                 77 
                 −16* 
                 77 
               
               
                 4 
                 25 
                 ug/ml 
                 14360 
                 18110 
                 64 
                 54 
                 59 
               
               
                 5 
                 12.5 
                 ug/m 
                 26570 
                 30474 
                 33 
                 23 
                 28 
               
               
                 6 
                 6.3 
                 ug/ml 
                 32559 
                 34113 
                 18 
                 14 
                 16 
               
               
                 7 
                 3.1 
                 ug/ml 
                 30136 
                 33881 
                 24 
                 14 
                 19 
               
               
                 8 
                 1.6 
                 ug/ml 
                 38674 
                 42196 
                 2 
                 −7 
                 −2 
               
               
                   
               
               
                 *Outliers 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 41 
               
             
            
               
                   
               
               
                 Dose-response curve 
               
               
                 Ayuric DRC 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 Conc. 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 1 
                 200 
                 ug/ml 
                 1510 
                 1974 
                 96 
                 95 
                 96 
               
               
                 2 
                 100 
                 ug/ml 
                 6546 
                 8141 
                 84 
                 80 
                 82 
               
               
                 3 
                 50 
                 ug/ml 
                 118831 
                 41081 
                 −202*  
                 −4 
                 −4 
               
               
                 4 
                 25 
                 ug/ml 
                 53443 
                 33759 
                 −36* 
                 14 
                 14 
               
               
                 5 
                 12.5 
                 ug/ml 
                 32062 
                 36771 
                 19 
                  7 
                 13 
               
               
                 6 
                 6.3 
                 ug/ml 
                 174800 
                 69322 
                 −344*  
                 −76* 
               
               
                 7 
                 3.1 
                 ug/ml 
                 80265 
                 49847 
                 −104*  
                 −26  
                 −26 
               
               
                 8 
                 1.6 
                 ug/ml 
                 34358 
                 38432 
                 13 
                  3 
                 8 
               
               
                   
               
               
                 *Outliers 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 42 
               
             
            
               
                   
               
               
                 Capros ® Dose-Response Curve 
               
               
                 Capros ® DRC 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 Conc. 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 1 
                 200 
                 ug/ml 
                 6929 
                 7493 
                 83 
                 81 
                 82 
               
               
                 2 
                 100 
                 ug/ml 
                 13583 
                 21326 
                 66 
                 46 
                 56 
               
               
                 3 
                 50 
                 ug/ml 
                 22529 
                 26420 
                 43 
                 33 
                 38 
               
               
                 4 
                 25 
                 ug/ml 
                 29369 
                 33909 
                 26 
                 14 
                 20 
               
               
                 5 
                 12.5 
                 ug/ml 
                 32722 
                 39366 
                 17 
                 0 
                 9 
               
               
                 6 
                 6.3 
                 ug/ml 
                 34111 
                 41002 
                 14 
                 −4 
                 5 
               
               
                 7 
                 3.1 
                 ug/ml 
                 33371 
                 41891 
                 15 
                 −6 
                 5 
               
               
                 8 
                 1.6 
                 ug/ml 
                 33010 
                 40669 
                 16 
                 −3 
                 7 
               
               
                   
               
            
           
         
       
     
       FIG. 21  shows, overall, that extracts of the present invention (Ayuflex®, Capros®, and Ayuric®) in combinations with each other showed better anti-cancer effects than extracts alone. Also, Ibrutinib showed a better inhibition profile for CLL cancer cells when tested in combination with these extracts than ibrutinib alone. Ibrutinib&#39;s effect can be enhanced with combined doses of extracts into subjects, according to the present invention. At the left side of  FIG. 21 , 3 uM ibrutinib alone inhibited CLL PDC proliferation by about 36%, however, in combination with Ayuflex® (30 ug/ml), Capros® (100 ug/ml) or Ayuric® (100 ug/ml), inhibition of proliferation increased to about 60% (Ayuflex®) or 66% (Capros®, Ayuric®). The inhibition of CLL cell proliferation by Ayuflex®, Capros®, or Ayuric® was previously measured at about 50% (Ayuflex®) and about 73-75% (Capros®, Ayuric®) (See right side of  FIG. 21 .) 
     Increasing the ibrutinib concentration to 10 uM increased the inhibition of CLL cell proliferation by ibrutinib alone by about 11%, to 41% inhibition of proliferation. In combination with Ayuflex® (30 ug/ml), Capros® (100 ug/ml) or Ayuric® (100 ug/ml), inhibition of proliferation of CLL cells changed to about 62% (Ayuflex®), 59% (Capros®), and 69% (Ayuric®). The highest level of inhibitory activity toward CLL cells shown in  FIG. 21  is shown by combinations of extracts of the present invention (Ayuflex® (30 ug/ml), Capros® (100 ug/ml) or Ayuric® (100 ug/ml)). The Ayuflex®/Capros® combination shown in  FIG. 21  inhibited proliferation of CLL cancer cells by 80%, while the Ayuflex®/Ayuric® combination and the Capros®/Ayuric® combination inhibited proliferation of the CLL PDCs by 88% and 90%, respectively. In contrast, as shown at the far right of  FIG. 21 , Ayuflex® (30 ug/ml) alone in a previous study inhibited CLL cell proliferation by 52%; Capros® (100 ug/ml), by 75%; and Ayuric® (100 ug/ml), by 73%. 10 uM doxorubicin showed relatively low inhibitory activity for CLL cells, as shown at the far right of  FIG. 21 . Table 43 shows data represented by  FIG. 21 . 
     
       
         
           
               
             
               
                 TABLE 43 
               
             
            
               
                   
               
               
                 Inhibition of CLL PDC proliferation 
               
            
           
           
               
               
            
               
                   
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                   
                 Avg 
                 SD 
                 n1 
                 n2 
                 n3 
                 Avg 
                 SD 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Ibrutinib @ 3 uM 
                 20689 
                 27455 
                 28014 
                 25386 
                 4077 
                 48 
                 31 
                 29 
                 36 
                 10 
               
               
                 Ayuflex @ 30 ug/ml + 
                 13369 
                 18253 
                 15428 
                 15683 
                 2452 
                 66 
                 54 
                 61 
                 60 
                 6 
               
               
                 Ibrutinib @ 3 uM 
               
               
                 Capros @ 100 ug/ml + 
                 11439 
                 14698 
                 14572 
                 13570 
                 1846 
                 71 
                 63 
                 63 
                 66 
                 5 
               
               
                 Ibrutinib @ 3 uM 
               
               
                 Ayuric @ 100 ug/ml + 
                 11783 
                 12997 
                 15371 
                 13384 
                 1825 
                 70 
                 67 
                 61 
                 66 
                 5 
               
               
                 Ibrutinib @ 3 uM 
               
               
                 Ibrutinib @ 10 uM 
                 19340 
                 27230 
                 173394* 
                 23285 
                 5579 
                 51 
                 31 
                 * 
                 41 
                 14 
               
               
                 Ayuflex @ 30 ug/ml + 
                 12542 
                 17995 
                 14819 
                 15119 
                 2739 
                 68 
                 55 
                 63 
                 62 
                 7 
               
               
                 Ibrutinib @ 10 uM 
               
               
                 Capros @ 100 ug/ml + 
                 18454 
                 16505 
                 13407 
                 16122 
                 2545 
                 53 
                 58 
                 66 
                 59 
                 6 
               
               
                 Ibrutinib @ 10 uM 
               
               
                 Ayuric @ 100 ug/ml + 
                 14344 
                 12561 
                  9754 
                 12220 
                 2314 
                 64 
                 68 
                 75 
                 69 
                 6 
               
               
                 Ibrutinib @ 10 uM 
               
               
                 Ayuflex @ 30 ug/ml + 
                 6094 
                 9211 
                  8212 
                 7839 
                 1592 
                 85 
                 77 
                 79 
                 80 
                 4 
               
               
                 Capros @ 100 ug/ml 
               
               
                 Ayuflex @ 30 ug/ml + 
                 5095 
                 5394 
                  4257 
                 4915 
                 589 
                 87 
                 87 
                 89 
                 88 
                 1 
               
               
                 Ayuric @ 100 ug/ml 
               
               
                 Capros @ 100 ug/ml + 
                 4269 
                 4344 
                  4088 
                 4234 
                 132 
                 89 
                 89 
                 90 
                 90 
                 0 
               
               
                 Ayuric @ 100 ug/ml 
               
               
                 Ayuflex @ 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
                 52 
                 13 
               
               
                 {previous study} 
               
               
                 Capros @ 100 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
                 75 
                 1 
               
               
                 {previous study} 
               
               
                 Ayuric @ 100 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
                 73 
                 8 
               
               
                 {previous study} 
               
               
                 Doxorubicin @ 10 uM 
                 33348 
                 43129 
                 39159 
                 38545 
                 4919 
                 16 
                 −9 
                  1 
                 2 
                 13 
               
               
                   
               
               
                 *Outliers 
               
            
           
         
       
     
       FIG. 22  represents information provided in Tables 44-47B, showing the inhibition of proliferation of acute myeloid leukemia (AML) PDCs in the anti-proliferation assay.  FIG. 22  compares the inhibition of proliferation of AML PDCs by the 6 different extracts in the anti-proliferation assay, with Sensoril® matching or outperforming anti-proliferative activity by drug standards GDC-0941 (pictilisib) and doxorubicin at all concentrations tested (100 ug/ml, 30 ug/ml, 10 ug/ml). PrimaVie® showed comparable activity to Sensoril® in its 100 ug/ml and 30 ug/ml concentrations. The PhytoBGS® 30 ug/ml concentration and the Ayuric® 10 ug/ml concentration approached the anti-proliferation effects shown by drug standard GDC-0941. Effects by AyuFlex® and Capros® are also depicted in  FIG. 22 . GDC-0941 (pictilisib) and doxorubicin are presented as standard drug controls at the far right of  FIG. 23 . 
     
       
         
           
               
             
               
                 TABLE 44 
               
             
            
               
                   
               
               
                 Assay Controls 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                   
                 Assay 
                   
               
               
                   
                 Avg 
                 SD 
                 fold 
                 Z′ 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 100% Proliferation (Cell Control)  
                 15453 
                 1102 
                 533 
                 0.78 
               
               
                 0% Proliferation (Media only) 
                 29 
                 10 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 45 
               
             
            
               
                   
               
               
                 Assay Drug Control 
               
               
                 GDC-0941 @ 10 uM 
               
            
           
           
               
               
            
               
                 Avg 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
               
                 6828 
                 5808 
                 56 
                 63 
                 59 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 46 
               
             
            
               
                   
               
               
                 Assay Drug Control 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
            
               
                 Avg 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
               
                 4686 
                 4147 
                 70 
                 73 
                 72 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 47A 
               
             
            
               
                   
               
               
                 Assay Results 
               
            
           
           
               
               
            
               
                   
                 % Inhibition 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex ® 
                 33 
                 −11 
                 11  
                 0 
                 −54  
                 −66  
                 −43 
                 −39  
               
               
                 Capros ® 
                 9 
                 −19 
                 46* 
                 19 
                 27 
                  25* 
                 −7 
                 −2 
               
               
                 Phyto-BGS ® 
                 33 
                 40 
                 74* 
                 61 
                 47 
                 32 
                 33 
                 20 
               
               
                 Primavie ® 
                 60 
                 51 
                 61  
                 67 
                  35* 
                  73* 
                 21 
                 28 
               
               
                 Sensoril ® 
                 71 
                 67 
                 82  
                 68 
                 57 
                 69 
                 51 
                  17* 
               
               
                 Ayuric ® 
                 41 
                 20 
                 41* 
                 8 
                  1 
                 52 
                 60 
                 36 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 47B 
               
             
            
               
                   
               
               
                 Assay Results 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 Avg % 
                   
                 Avg % 
                   
                 Avg % 
                   
               
               
                   
                 inhibition 
                 SD 
                 inhibition 
                 SD 
                 inhibition 
                 SD 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Ayuflex ® 
                 33 
                 0 
                 6 
                 8 
                 −41 
                 3 
               
               
                 Capros ® 
                 9 
                 0 
                 23 
                 6 
                 −5 
                 4 
               
               
                 Phyto-BGS ® 
                 37 
                 5 
                 54 
                 10 
                 28 
                 7 
               
               
                 Primavie ® 
                 56 
                 7 
                 64 
                 4 
                 25 
                 5 
               
               
                 Sensoril ® 
                 69 
                 2 
                 69 
                 13 
                 60 
                 13 
               
               
                 Ayuric ® 
                 30 
                 15 
                 5 
                 5 
                 44 
                 11 
               
               
                   
               
            
           
         
       
     
     As Phyto-BGS®, PrimaVie®, and Sensoril® showed the highest anti-cancer, anti-proliferative activity for AML PDCs of the extracts tested in  FIG. 22 , testing with the aim of revealing a dose-response curve and IC 50  for AML cancer cells was performed.  FIG. 23  shows dose-response curves for Phyto-BGS®, PrimaVie®, and Sensoril® on AML cancer cells. An IC 50  of about 19.51 ug Sensoril®/ml was measured. Sensoril® showed the best anti-cancer activity of the 3 extracts tested. In a previous study, Sensoril® showed 60% inhibition at a 10 ug/ml dose, when tested at 3 graded doses. Tables 48-50 show data used to generate the dose-response curves. 
     
       
         
           
               
             
               
                 TABLE 48 
               
             
            
               
                   
               
               
                 Dose-response curve 
               
               
                 Phyto-BGS DRC 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 Conc. 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 1 
                 200 
                 ug/ml 
                 6031 
                 8522 
                 63 
                 48 
                 55 
               
               
                 2 
                 100 
                 ug/ml 
                 11057 
                 14285 
                 32 
                 12 
                 22 
               
               
                 3 
                 50 
                 ug/ml 
                 12265 
                 15660 
                 25 
                 4 
                 14 
               
               
                 4 
                 25 
                 ug/ml 
                 13697 
                 16313 
                 16 
                 0 
                 8 
               
               
                 5 
                 12.5 
                 ug/ml 
                 13577 
                 16924 
                 17 
                 −4 
                 6 
               
               
                 6 
                 6.3 
                 ug/ml 
                 14206 
                 17655 
                 13 
                 −8 
                 2 
               
               
                 7 
                 3.1 
                 ug/ml 
                 14013 
                 15451 
                 14 
                 5 
                 10 
               
               
                 8 
                 1.6 
                 ug/ml 
                 12746 
                 14484 
                 22 
                 11 
                 16 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 49 
               
             
            
               
                   
               
               
                 Dose-response curve 
               
               
                 Primavie DRC 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 Conc. 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 1 
                 200 
                 ug/ml 
                 11209 
                 14738 
                 31 
                 10 
                 20 
               
               
                 2 
                 100 
                 ug/ml 
                 14520 
                 18950 
                 11 
                 −16 
                 −3 
               
               
                 3 
                 50 
                 ug/ml 
                 13778 
                 17755 
                 15 
                 −9 
                 3 
               
               
                 4 
                 25 
                 ug/ml 
                 14414 
                 16829 
                 12 
                 −3 
                 4 
               
               
                 5 
                 12.5 
                 ug/ml 
                 14095 
                 15965 
                 14 
                 2 
                 8 
               
               
                 6 
                 6.3 
                 ug/ml 
                 13534 
                 16727 
                 17 
                 −3 
                 7 
               
               
                 7 
                 3.1 
                 ug/ml 
                 13415 
                 16638 
                 18 
                 −2 
                 8 
               
               
                 8 
                 1.6 
                 ug/ml 
                 15363 
                 17617 
                 6 
                 −8 
                 −1 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 50 
               
             
            
               
                   
               
               
                 Dose-response curve 
               
               
                 Sensoril DRC 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 Conc. 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 1 
                 200 
                 ug/ml 
                 238 
                 344 
                 99 
                 98 
                 98 
               
               
                 2 
                 100 
                 ug/ml 
                 474 
                 740 
                 97 
                 96 
                 96 
               
               
                 3 
                 50 
                 ug/ml 
                 3667 
                 3860 
                 78 
                 76 
                 77 
               
               
                 4 
                 25 
                 ug/ml 
                 6173 
                 7312 
                 62 
                 55 
                 59 
               
               
                 5 
                 12.5 
                 ug/ml 
                 8103 
                 9989 
                 50 
                 39 
                 45 
               
               
                 6 
                 6.3 
                 ug/ml 
                 10181 
                 11777 
                 38 
                 28 
                 33 
               
               
                 7 
                 3.1 
                 ug/ml 
                 12124 
                 14101 
                 26 
                 14 
                 20 
               
               
                 8 
                 1.6 
                 ug/ml 
                 13339 
                 15317 
                 18 
                 6 
                 12 
               
               
                   
               
            
           
         
       
     
       FIG. 24  represents information provided in Table 51. Phyto-BGS® (30 ug/ml) in combination with Arsenic Trioxide (3 uM), Cytarabine (3 uM), or Doxorubicin (3 uM), inhibited AML proliferation by 86%, 97%, and 100%, respectively. PrimaVie® (30 ug/ml) in combination with Arsenic Trioxide (3 uM), Cytarabine (3 uM), or Doxorubicin (3 uM), inhibited AML proliferation by 82%, 96%, and 100%, respectively. Sensoril® (10 ug/ml) in combination with Arsenic Trioxide (3 uM), Cytarabine (3 uM), or Doxorubicin (3 uM), inhibited AML proliferation by 90%, 96%, and 100%, respectively. The known anti-cancer drugs tested—Arsenic Trioxide, Cytarabine, and Doxorubicin, are shown as inhibiting AML proliferation by 97%, 99%, and 100%. Surprisingly, in combination with each other, the extracts tested performed well when compared the known anti-cancer drugs used as controls. The combination of Phyto-BGS (30 ug/ml) and PrimaVie® (30 ug/ml) inhibited AML cell proliferation by 85%; the Phyto-BGS (30 ug/ml) and Sensoril® (10 ug/ml) combination inhibited AML cell proliferation by 97%; and the PrimaVie® (30 ug/ml) and Sensoril® combination inhibited AML cell proliferation by 100%. In a previous study Phyto-BGS® (100 ug/ml) inhibited AML cell proliferation by 37%; PrimaVie® (100 ug/ml) inhibited AML cell proliferation by 56%; and Sensoril® (10 ug/ml) inhibited AML cell proliferation by 60%. Overall, in  FIG. 24 , extracts in combinations showed better anti-cancer effects than extracts alone. Known AML drugs cytarabine, doxorubicin, and arsenic trioxide (each at 3 uM) alone showed greater than 95% inhibition (97-100%), whereas combinations of extracts of the present invention showed AML cell proliferation inhibition at or greater than 85% (85-100%). 
     
       
         
           
               
             
               
                 TABLE 51 
               
             
            
               
                   
               
               
                 Inhibition of AML proliferation 
               
            
           
           
               
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 Avg 
                 SD 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Phyto-BGS ®@ 30 ug/ml 
                 11231 
                 13737 
                 12743 
                   35**   
                 21 
                 27 
                 24 
                 4.0 
               
               
                 Phyto-BGS ®@30 ug/ml + 
                 1993 
                 2545 
                 2608 
                 89 
                 85 
                 85 
                 86 
                 1.9 
               
               
                 Arsenic Trioxide@ 3 uM 
               
               
                 Phyto-BGS ®@ 30 ug/ml + 
                 489 
                 635 
                 586 
                 97 
                 96 
                 97 
                 97 
                 0.4 
               
               
                 Cytarabine @ 3 uM 
               
               
                 Phyto-BGS ®@ 30 ug/ml + 
                 56 
                 39 
                 49 
                 100  
                 100 
                 100 
                 100 
                 0.0 
               
               
                 Doxorubicin @ 3 uM 
               
               
                 Primavie ®@ 30 ug/ml 
                 10627 
                 13790 
                 14651 
                   39*   
                 21 
                 16 
                 18 
                 3.5 
               
               
                 Primavie ®@ 30 ug/ml + 
                 2766 
                 3463 
                 3273 
                 84 
                 80 
                 81 
                 82 
                 2.1 
               
               
                 Arsenic Trioxide @ 3 uM 
               
               
                 Primavie ®@ 30 ug/ml + 
                 686 
                 690 
                 731 
                 96 
                 96 
                 96 
                 96 
                 0.1 
               
               
                 Cytarabine @ 3 uM 
               
               
                 Primavie ®@ 30 ug/ml + 
                 41 
                 66 
                 45 
                 100  
                 100 
                 100 
                 100 
                 0.1 
               
               
                 Doxorubicin @ 3 uM 
               
               
                 Sensoril ®@ 10 ug/ml 
                 7146 
                 9782 
                 10670 
                   59*   
                 44 
                 39 
                 41 
                 3.6 
               
               
                 Sensoril ®@ 10 ug/ml + 
                 1470 
                 2007 
                 1796 
                 92 
                 89 
                 90 
                 90 
                 1.6 
               
               
                 Arsenic Trioxide @ 3 uM 
               
               
                 Sensoril ®@ 10 ug/ml + 
                 588 
                 731 
                 835 
                 97 
                 96 
                 95 
                 96 
                 0.7 
               
               
                 Cytarabine @ 3 uM 
               
               
                 Sensoril ®@ 10 ug/ml + 
                 34 
                 33 
                 42 
                 100  
                 100 
                 100 
                 100 
                 0.0 
               
               
                 Doxorubicin @ 3 uM 
               
               
                 Arsenic Trioxide@ 3 uM 
                 551 
                 599 
                 484 
                 97 
                 97 
                 97 
                 97 
                 0.3 
               
               
                 Cytarabine@ 3 uM 
                 203 
                 192 
                 140 
                 99 
                 99 
                 99 
                 99 
                 0.2 
               
               
                 Doxorubicin@ 3 uM 
                 25 
                 45 
                 32 
                 100  
                 100 
                 100 
                 100 
                 0.1 
               
               
                 PhytoBGS ®@30 ug/ml + 
                 2545 
                 2857 
                 2390 
                 85 
                 84 
                 86 
                 85 
                 1.4 
               
               
                 Primavie ®@30 ug/ml 30 ug/l 
               
               
                 PhytoBGS ®@30 ug/ml + 
                 557 
                 583 
                 534 
                 97 
                 97 
                 97 
                 97 
                 0.1 
               
               
                 Sensoril ®@ 10 ug/ml 
               
               
                 Primavie ®@30 ug/ml + 
                 36 
                 37 
                 39 
                 100  
                 100 
                 100 
                 100 
                 0.0 
               
               
                 Sensoril ®@ 10 ug/ml 
               
               
                   
               
               
                 Bolded text = Outliers 
               
            
           
         
       
     
     Example II 
     Inhibition of Acute Myeloid Leukemia Cell Proliferation by Sensoril® 
     AML (Acute myeloid leukemia) is one of the most common types of leukemia in adults. AML is an aggressive disease in which numerous myeloblasts are found in the bone marrow and blood of a subject. AML can spread to other parts of the body including the lymph nodes, liver, spleen, central nervous system, and testicles. 
     Several drugs have been approved to treat AML by the US Food and Drug Administration and are available to subjects in need, however, these drugs may have less than desired efficacy and may produce undesirable side, adverse, or toxic effects. Examples of drugs currently available to treat AML include arsenic trioxide (As 2 O 3 ), cytarabine, and doxorubicin. Current therapeutic targets for treating AML include inhibitors of FLT3, IDH, histone deacetylase (HDAC), and BCL-2 polo-like kinase (Plk). 
     Anti-cancer luminescence assays on AML primary cells included incubation with extract for 72 hours, as in Example I. Doxorubicin was used as an assay control. In this Example, AML drugs were used as disease specific controls. 
     As discussed above in Example I, 6 extracts (Ayuflex®, Capros®, Phyto-BGS®, PrimaVie®, Sensoril®, and Ayuric®) were tested on AML PDC-SB cells (33866, M-4 type, 76 year old subject) at 3 concentrations: 100 ug/ml, 30 ug/ml, and 10 ug/ml. Three extracts (Phyto-BGS®, PrimaVie®, and Sensoril®) showed desirable activity in those tests and were further tested on the same PDCs (patient-derived cells) for determination of their IC 50  values and in combination with marketed AML drugs such as cytarabine, doxorubicin, or arsenic trioxide or in combination with other extracts. Sensoril® showed the best and most remarkable anti-cancer anti-proliferative activity alone with 60% inhibition at 10 ug/ml, tested at 3 graded doses, and with an IC 50  of about 20 ug/ml (See  FIG. 23 , showing an IC 50  of 19.51 ug/ml). The plate passed quality control criteria with a Z′ of 0.63. Extracts in combinations showed synergistic anti-cancer effects compared with extracts alone. AML drugs cytarabine, doxorubicin, and arsenic trioxide alone at 3 uM concentrations showed greater than 95% inhibition. See for instance  FIGS. 1-6 and 22-24  and associated Tables and related discussion above.  FIG. 22  shows anti-cancer effects of Phyto-BGS®), PrimaVie®, and Sensori)®;  FIG. 23  shows dose-response curves for these 3 extracts on the AML PDCs, and  FIG. 24  shows efficacy of the extracts and combinations as discussed above. These studied showed in part 37% inhibition by Phyto-BGS® (100 ug/ml), 56% inhibition by PrimaVie® (100 ug/ml), and 60% inhibition by Sensoril® (10 ug/ml). 
     The data in Tables 52-59 and  FIGS. 25-28  is from experiments using different AML PDCs than those discussed above to show Sensoril&#39;s effects on other PDCs of a different AML subtype. The AML PDCs evaluated below were an Acute Promyelocytic Leukemia (APL, APML) subtype, APL-SB 46120, from a 15-year-old human subject. 
     Sensoril® was evaluated at 3 different concentrations (IC 75 , IC 50 , and IC 25 , derived from information such as shown in  FIG. 23  and related information above), and also evaluated with IC 50  and IC 25  concentrations of marketed drugs, in part to show potential combinations that provide improved efficacy when administered to subjects. As the concentration of cytarabine, doxorubicin, and arsenic trioxide showed greater than 95% inhibition at 3 uM concentrations in Example I, lowering the concentrations of these drugs enabled a clearer showing of the synergistic effect of Sensoril®. In one experiment, the IC 50  and IC 25  of cytarabine, doxorubicin, and arsenic trioxide on the M4 APL cells discussed above and for instance in  FIGS. 22-24  was determined. IC 25  values were calculated using IC 50  and hill slope. The IC 50  and IC 25  values of all three AML drugs on the M4 cells are described in Table 52. The drugs were used in these concentrations on the new APL subtype cells as described further below. Also, a dose-response curve for Sensoril® was generated in the new APL subtype PDCs, as shown in  FIG. 25 , and an IC 50  of 32.94 ug/ml was determined for the APL subtype PDCs. As a comparison and as noted above, the IC 50  for the M4 cells in previous studies was 19.5 ug/ml. Tables 53-57 present quality control and data analysis information relating to these experiments. 
     
       
         
           
               
             
               
                 TABLE 52 
               
             
            
               
                   
               
               
                 Inhibitory Concentrations of Sensoril ® and AML drugs on AML cells 
               
            
           
           
               
               
               
               
               
            
               
                 Inhibitory 
                   
                   
                   
                   
               
               
                 Concentration 
                   
                   
                   
                   
               
               
                 (IC) 
                 Sensoril ® 
                 Cytarabine 
                 As 2 O 3   
                 Doxorubicin 
               
               
                   
               
               
                 IC 25   
                  8.2 ug/ml 
                  23 nM 
                 1.38 uM 
                  40.7 nM 
               
               
                 IC 50   
                  19.5 ug/ml 
                 104 nM 
                 2.99 uM 
                 102.8 nM 
               
               
                 IC 75   
                 46.25 ug/ml 
                 470 nM 
                 6.45 uM 
                   260 nM 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 53 
               
             
            
               
                   
               
               
                 Plate 1 Quality Control 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 Standard 
                   
                   
               
               
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 100% Proliferation 
                 11591 
                 1061 
                 169 
                 0.72 
               
               
                 (Cell Control) 
                   
                   
                   
                   
               
               
                 0% Proliferation 
                 69 
                 5 
                   
                   
               
               
                 (Media only) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 54 
               
             
            
               
                   
               
               
                 Plate 1 Quality Control (Doxorubicin) 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                 Average 
                 % Inhibition 
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 Average 
               
               
                   
               
               
                 465 
                 486 
                 627 
                 97 
                 96 
                 95 
                 96 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 55 
               
             
            
               
                   
               
               
                 Plate 2 Quality Control 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                 Standard 
                   
                   
               
               
                   
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 100% Proliferation 
                 16734 
                 1146 
                 167 
                 0.79 
               
               
                   
                 (Cell Control) 
                   
                   
                   
                   
               
               
                   
                 0% Proliferation 
                 101 
                 35 
                   
                   
               
               
                   
                 (Media only) 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 56 
               
             
            
               
                   
               
               
                 Plate 2 Quality Control (Doxorubicin) 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                 Average 
                 % Inhibition 
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 Average 
               
               
                   
               
               
                 418 
                 668 
                 621 
                 98 
                 97 
                 97 
                 97 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 57 
               
             
            
               
                   
               
               
                 Plate 1 - Sensoril ® Dose-Response Curve 
               
               
                 Sensoril ® Dose-Response Curve (DRC) (IC 50  = 32.94 ug/ml) 
               
            
           
           
               
               
               
               
            
               
                   
                 Concentration 
                 Raw data 
                 Analysis 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 Sample 
                 ug/ml 
                 n1 
                 n2 
                 n1 
                 n2 
                 Average 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 1 
                 200 
                 2617 
                 2926 
                 78 
                 75 
                 77 
                 2 
               
               
                 2 
                 100 
                 3579 
                 4065 
                 70 
                 65 
                 67 
                 3 
               
               
                 3 
                 50 
                 4661 
                 5182 
                 60 
                 56 
                 58 
                 3 
               
               
                 4 
                 25 
                 7733 
                 10052 
                 33 
                 13 
                 23 
                 14 
               
               
                 5 
                 12.5 
                 9878 
                 11050 
                 15 
                 5 
                 10 
                 7 
               
               
                 6 
                 6.25 
                 12217 
                 13830 
                 −5 
                 −19 
                 −12 
                 10 
               
               
                 7 
                 3.13 
                 9193 
                 10849 
                 21 
                 6 
                 14 
                 10 
               
               
                 8 
                 1.56 
                 12252 
                 14753 
                 −6 
                 −27 
                 −17 
                 15 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 58 
               
             
            
               
                   
               
               
                 Plate 1 
               
            
           
           
               
               
               
            
               
                 Combination 
                 Avg 
                 SD 
               
               
                   
               
            
           
           
               
               
               
            
               
                 Sensoril IC75 
                 37 
                 7 
               
               
                 Sensoril IC50 
                 −13 
                 12 
               
               
                 Sensoril IC25 
                 −7 
                 20 
               
               
                 Sensoril IC75 + Cytarabine IC50 
                 36 
                 4 
               
               
                 Sensoril IC50 + Cytarabine IC50 
                 −26 
                 15 
               
               
                 Sensoril IC25 + Cytarabine IC50 
                 −20 
                 27 
               
               
                 Sensoril IC75 + Cytarabine IC25 
                 46 
                 13 
               
               
                 Sensoril IC50 + Cytarabine IC25 
                 −19 
                 15 
               
               
                 Sensoril IC25 + Cytarabine IC25 
                 −62 
                 7 
               
               
                 Cytarabine IC50 
                 20 
                 16 
               
               
                 Cytarabine IC25 
                 −51 
                 15 
               
               
                 Doxorubin @ 10 uM 
                 96 
                 1 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 59 
               
             
            
               
                   
               
               
                 Plate 2 
               
               
                 Plate 2 
               
            
           
           
               
               
               
               
            
               
                   
                 Combination 
                 Avg 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Sensoril IC75 + As203 IC50 
                 90 
                 2 
               
               
                   
                 Sensoril IC50 + As203 IC50 
                 94 
                 1 
               
               
                   
                 Sensoril IC25 + As203 IC50 
                 93 
                 2 
               
               
                   
                 Sensoril IC75 + As203 IC25 
                 93 
                 1 
               
               
                   
                 Sensoril IC50 + As203 IC25 
                 94 
                 3 
               
               
                   
                 Sensoril IC25 + As203 IC25 
                 94 
                 1 
               
               
                   
                 Sensoril IC75 + Doxorubin IC50 
                 93 
                 3 
               
               
                   
                 Sensoril IC50 + Doxorubin IC50 
                 98 
                 2 
               
               
                   
                 Sensoril IC25 + Doxorubin IC50 
                 98 
                 1 
               
               
                   
                 Sensoril IC75 + Doxorubin IC25 
                 97 
                 2 
               
               
                   
                 Sensoril IC50 + Doxorubin IC25 
                 93 
                 1 
               
               
                   
                 Sensoril IC25 + Doxorubin IC25 
                 94 
                 2 
               
               
                   
                 AS203 IC50 
                 60 
                 9 
               
               
                   
                 AS203 IC25 
                 16 
                 14 
               
               
                   
                 Doxorubin IC50 
                 43 
                 18 
               
               
                   
                 Doxorubin IC25 
                 −41 
                 5 
               
               
                   
                 Doxorubin 10 uM 
                 97 
                 1 
               
               
                   
               
            
           
         
       
     
     In  FIGS. 26-28 , Sensoril® was evaluated at 3 different concentrations alone (IC 75 , IC 50 , IC 25 ; Table 52), and combined with IC 50  and IC 25  concentrations of AML drugs (see also Tables 58-59). Sensoril® showed additive and synergistic anti-cancer anti-proliferative effects when combined with As 2 O 3  ( FIG. 27 ) and Doxorubicin ( FIG. 28 ). Sensoril® showed similar IC 50  values for the AML subtype APL PDCs (approximately 31 ug/ml) and the AML (M4) PDCs of Example 1 (approximately 20 ug/ml). Arsenic trioxide and doxorubicin alone showed good efficacy compared with cytarabine alone. These results tally with clinical treatment, as arsenic trioxide and doxorubicin are preferred drugs for APL patients. In this study, Sensoril® in combination with cytarabine showed some but no significant inhibition of the subtype APL PDCs ( FIG. 26 ). 
     Sensoril® shows potent anti-cancer anti-proliferative activity against different subtypes of AML, alone and in combination with drugs relevant to AML and APL treatment. Sensoril® may be administered with standard AML drugs, including at a low dose of the AML drugs, to achieve optimal efficacy and lower side effects of the drugs. 
     Example III 
     Inhibition of Non-Small Cell Lung Cancer Cell and Colon Cancer Cell Proliferation with  Terminalia chebula  (AyuFlex®) and  Terminalia bellerica  (Ayuric®) 
     Anti-proliferative and anti-cancer activity of extracts of this invention on non-small cell lung cancer cells and colon cancer cells were assessed generally as described in Example I and are discussed below.  Terminalia chebula  (AyuFlex®) and  Terminalia bellerica  (Ayuric®) showed significant activity. As noted throughout this application, the below data is intended to describe the present invention but not to be limiting. 
     Non-small cell lung cancer cells (cell line NCI-H-358) and colon cancer cells (cell line HT-29) were plated in a 384-well plate in amounts of about 1.25 k cells/well. AyuFlex®, Capros®, Phyto-BGS®, Primavie®, Sensoril®, Ayuric®, and Crominex® were added to the cells in concentrations of 100 ug/ml, 30 ug/ml, and 10 ug/ml, in triplicate, and incubated for 72 hours, and CellTiterGlo (Promega Corporation, Wisconsin, USA) used as a detection reagent, as described in Example I. Drugs used to treat non-small cell lung cancer (Standard of Care (SOC) drugs) Docetaxel (10 uM) and Doxorubicin (10 uM) were used as controls, as were drugs used to treat colon cancer (Standard of Care (SOC) drugs) 5-FU (5-fluoro-uracil, 10 uM) and Doxorubicin (10 uM). The IC 50  of docetaxel for non-small cell lung cancer H-358 cells has been published as 367 nM ( Intl. J. Oncol.  31:241-252 (2007)). The IC 50  of 5-FU for colon cancer H-29 cells has been published as 13 uM (J. Surg. Res. 111(1):63-69 (2003)). Wells with untreated cells and with plain media were also included as controls. Plates passed quality control parameters. 
     Anti-Cancer Anti-Proliferative Inhibition of Non-Small Cell Lung Cancer Cells 
     Tables 60-62 show data for controls, and  FIG. 29  shows data relating to the 7 extracts tested (AyuFlex®, Capros®, Phyto-BGS®, Primavie®, Sensoril®, Ayuric®, and Crominex®), with bars reading left to right and the associated table reading top to bottom: 100 ug/ml, 30 ug/ml, and 10 ug/ml. Docetaxel exhibited 58% inhibition of non-small cell lung cancer cell proliferation at a 10 uM concentration. Assay control doxorubicin at 10 uM showed 78% inhibition of non-small cell lung cancer cell proliferation at a 10 uM concentration. 
     Of the 7 extracts tested and shown in  FIG. 29  (left to right: Ayuflex®, Capros®, Phyto-BGS®, Primavie®, Sensoril®, Ayuric®, Crominex®, at 100, 30, and 10 ug/ml concentrations), Ayuric® showed the most potent anti-cancer and anti-proliferative effect, with 91% inhibition of proliferation at a 100 ug/ml concentration and 27% inhibition at 30 ug/ml. Ayuflex® also showed an anti-cancer anti-proliferative effect, inhibiting proliferation of non-small cell lung cancer cells by 40% at a 100 ug/ml concentration. The other extracts tested showed a lesser or no effect on cells of this lung cancer cell line. 
     
       
         
           
               
             
               
                 TABLE 60 
               
             
            
               
                   
               
               
                 Plate Controls 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 Standard 
                   
                   
               
               
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 100% Proliferation 
                 62761 
                 5395 
                 1111 
                 0.74 
               
               
                 (Cell Control) 
                   
                   
                   
                   
               
               
                 0% Proliferation 
                 57 
                 18 
                   
                   
               
               
                 (Media only) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 61 
               
             
            
               
                   
               
               
                 Plate Controls - Docetaxel 
               
               
                 Docetaxel @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 23464 
                 28364 
                 26690 
                 63 
                 55 
                 58 
                 58(4) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 62 
               
             
            
               
                   
               
               
                 Plate Controls - Doxorubicin 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 13222 
                 14367 
                 14575 
                 79 
                 77 
                 77 
                 78(1) 
               
               
                   
               
            
           
         
       
     
     Anti-Cancer Anti-Proliferative Inhibition of Colon Cancer Cells 
     Tables 63-65 show data for controls and that the data passed quality control parameters.  FIG. 30  shows data relating to the 7 extracts tested (AyuFlex®, Capros®, Phyto-BGS®, Primavie®, Sensoril®, Ayuric®, and Crominex®), with bars reading left to right and the associated table reading top to bottom: 100 ug/ml, 30 ug/ml, and 10 ug/ml. 5-FU provided only 12% inhibition of colon cancer cell proliferation at a 10 uM concentration. Assay control doxorubicin showed inhibition of colon cancer cell proliferation by 94%. 
     Of the 7 extracts tested, Ayuric® showed the most potent anti-cancer anti-proliferative activity, providing 80% inhibition of colon cancer cell proliferation at a concentration of 100 ug/ml and 22% inhibition at 30 ug/ml. Ayuflex® inhibited colon cancer cell proliferation by 45% at a 100 ug/ml concentration. The remaining 5 extracts showed little to no effect on this colon cancer cell line. 
     
       
         
           
               
             
               
                 TABLE 63 
               
             
            
               
                   
               
               
                 Plate Controls 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                 Standard 
                   
                   
               
               
                   
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 100% Proliferation 
                 92463 
                 2167 
                 1988 
                 0.93 
               
               
                   
                 (Cell Control) 
                   
                   
                   
                   
               
               
                   
                 0% Proliferation 
                 47 
                 4 
                   
                   
               
               
                   
                 (Media only) 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 64 
               
             
            
               
                   
               
               
                 Plate Controls - 5-FU 
               
               
                 5-FU @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 74375 
                 86123 
                 83116 
                 20 
                 7 
                 10 
                 12(7) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 65 
               
             
            
               
                   
               
               
                 Plate Controls - Doxorubicin 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 5430 
                 6159 
                 5802 
                 94 
                 93 
                 94 
                 94(0) 
               
               
                   
               
            
           
         
       
     
     Example IV 
     Inhibition of Glioma Cells with AyuFlex®, Capros®, and Ayuric® 
     As discussed with regard to  FIG. 7  in Example I above, AyuFlex®, Capros®, and Ayuric® showed significant dose-dependent inhibition of patient derived primary glioma cells (described in Table 1). Also, as shown in  FIG. 8 , AyuFlex®, Capros®, and Ayuric® exhibited better inhibition than a standard-of-care drug, temozolomide, when tested alone or in combination. Combinations of these extracts (AyuFlex®+Capros®, AyuFlex®+Ayuric®, Capros®+Ayuric®) exhibited potent anti-cancer activity in the glioma PDCs, with &gt;73% inhibition. 
     The anti-cancer activity of AyuFlex®, Capros®, and Ayuric® was further tested on two other glioma PDCs taken from 2-3 different subjects (SB 6129, SB 32833 (55 year old male)). These glioma PDCs had a different mix of cell types (neuronal, astrocytic, dendroglioma). Also, the anti-cancer activity of AyuFlex®, Capros®, and Ayuric® was further tested on existing glioblastoma cell line U87 MG, to allow comparisons with other compositions known in the art. AyuFlex®, Capros®, and/or Ayuric® were incubated with the new PDC lines and the U87 MG cell line for 72 hours, as described in Example I above. 
     Tables 66-67 show data for controls and that the data passed quality control parameters. Table 68 relates to  FIG. 31 , a graph showing the efficacy of AyuFlex®, Capros®, and/or Ayuric® on Glioma SB 32833 Glioblastoma Grade IV patient derived cells, in a combination study. The graph shows, left to right, % inhibition of SB 32833 PDCs by AyuFlex® (100, 30, 10 ug/ml); Capros® (100, 30, 10 ug/ml); and Ayuric® (100, 30, 10 ug/ml); as well as combinations AyuFlex® (100 ug/ml)+Capros® (100 ug/ml), AyuFlex® (100 ug/ml)+Ayuric® (100 ug/ml), AyuFlex® (30 ug/ml)+Capros® (100 ug/ml), AyuFlex® (30 ug/ml)+Ayuric® (100 ug/ml), and Capros® (100 ug/ml)+Ayuric (100 ug/ml). (GDC-0941@10 uM, Doxorubicin@10 uM, and Temozolomide@100 uM shown at far right).  FIG. 32  also shows the efficacy of the above extract doses (same as for  FIG. 31 ) and combinations on glioma SB 6129 (anaplastic astrocytoma Grade III PDCs.  FIG. 33  shows the efficacy of AyuFlex®, Capros®, and Ayuric® (same extract doses as  FIG. 31 ) after a 72 hour incubation on known cell line U87-MG (glioblastoma) A dose-dependent effect was observed for all 3 extracts, with the anti-cancer effect on SB 32833 PDCs comparable to that described in Example 1 including  FIGS. 7-8  above. 
     
       
         
           
               
             
               
                 TABLE 66 
               
             
            
               
                   
               
               
                 Control data 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                 Standard 
                   
                   
               
               
                   
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 100% Proliferation 
                 39636 
                 4762 
                 855 
                 0.64 
               
               
                   
                 (Cell Control) 
                   
                   
                   
                   
               
               
                   
                 0% Proliferation 
                 46 
                 25 
                   
                   
               
               
                   
                 (Media only) 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 67 
               
             
            
               
                   
               
               
                 Doxorubicin control 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 3578 
                 5759 
                 5708 
                 91 
                 86 
                 86 
                 87 
               
               
                   
               
            
           
         
       
     
                     TABLE 68                  Inhibition of Glioma Cells                                 Raw data   % Inhibition                                                     Combination   n1   n2   n3   n1   n2   n3   Avg   SD                                                         Ayuflex 100 ug/ml   4306   5649   5809   89   86   85   87   2       Ayuflex 30 ug/ml   8975   11126   12150   77   72   69   73   4       Ayuflex 10 ug/ml   17236   23956   23625   57   40   40   46   10       Capros 100 ug/ml   10643   13866   12564   73   65   68   69   4       Capros 30 ug/ml   21842   29142   29043   45   27   27   33   11       Capros 10 ug/ml   27354   36276   35965   31   8   9   16   13       Ayuric 100 ug/ml   5066   7275   7389   87   82   81   84   3       Ayuric 30 ug/ml   14419   20278   18373   64   49   54   55   8       Ayuric 10 ug/ml   15383   19439   19324   61   51   51   55   6       Ayuflex 100 ug/ml +   3083   4733   5300   92   88   87   89   3       Capros 100 ug/ml                                       Ayuflex 100 ug/ml +   474   1034   589   99   98   99   98   1       Ayuric 100 ug/ml                                       Ayuflex 30 ug/ml +   5109   8377   8192   87   79   79   82   5       Capros 100 ug/ml                                       Ayuflex 30 ug/ml +   4701   5847   5574   88   85   86   87   2       Ayuric 100 ug/ml                                       Capros 100 ug/ml +   15   15   20   100   100   100   100   0       Ayuric 100 ug/ml                                       GDC-0941 @ 10 uM   20   16   16   100   100   100   100   0       Doxorubicin @ 10 uM   3578   5759   5708   91   86   86   87   3       Temozolomide @ 100 uM   23511   31320   32454   41   21   18   27   12                    
In the three independent studies using 3 different gliomas (1 grade III and 2 grade IV gliomas), AyuFlex®, Capros®, and/or Ayuric® alone and in combination exhibited significant anti-cancer effects. Glioblastoma multiforme (GBM) is one of the most challenging brain tumor to treat, as patients generally do not live more than 1-2 years. Temozolomide is one of the only approved treatments, or the only treatment, for gliomas, however, 50-70% of patients treated with temozolomide have been reported as non-responders to temozolomide. Accordingly, the present invention includes treatment with AyuFlex®, Capros®, and/or Ayuric®, or other compositions of the present invention.
 
     Example V 
     Inhibition of AML Cells with Sensoril®, Phyto-BGS, and PrimaVie® 
     As shown in  FIGS. 22, 24, 26, 27, and 28 , Sensoril®, Phyto-BGS® and PrimaVie® compositions of this invention inhibited 2 different types of AML PDCs. As shown in this Example, Sensoril®, Phyto-BGS®, and PrimaVie® compositions, and also a Crominex® composition, inhibited cancer cell proliferation in an AML cell line: HL60. 
     Inhibition assays were generally run as described for other AML assays above, with 5000 cells per well in 384 well plates. Standard-of-care drugs cytarabine (3 uM), arsenic trioxide (As 2 O 3 ) (60 uM) and doxorubicin (10 uM) were assayed as controls. Phyto-GBS®, PrimaVie®, and Crominex® were assayed at 100 ug/ml, 30 ug/ml, and 10 ug/ml concentrations, and Sensoril® tested with a 9-point, 2-fold dose response curve (DRC), starting from 200 ug/ml. CellTiterGlo detection reagent was used, as discussed for instance in Example I above, and the 4 compositions were incubated with the cells for 72 hours. Tables 69-70 show data for controls and that the data passed quality control parameters. 
     
       
         
           
               
             
               
                 TABLE 69 
               
             
            
               
                   
               
               
                 Assay controls—Plate 1 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 Standard 
                   
                   
               
               
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 100% Proliferation 
                 43535 
                 2633 
                 2561 
                 0.82 
               
               
                 (Cell Control) 
                   
                   
                   
                   
               
               
                 0% Proliferation 
                 17 
                 1 
                   
                   
               
               
                 (Media only) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 70 
               
             
            
               
                   
               
               
                 Assay controls—Plate 2 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 Standard 
                   
                   
               
               
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 100% Proliferation 
                 87067 
                 5601 
                 189 
                 0.81 
               
               
                 (Cell Control) 
                   
                   
                   
                   
               
               
                 0% Proliferation 
                 461 
                 18 
                   
                   
               
               
                 (Media only) 
               
               
                   
               
            
           
         
       
     
     As shown in  FIG. 34 , Phyto-BGS® and PrimaVie® showed dose-dependent anti-cancer effects on HL60 cells, with greater than 60% inhibition at a dose of 100 ug/ml. Crominex® demonstrated 30-40% inhibition at all doses (100, 30, 10 ug/ml). Control standard-of-care drugs cytarabine (3 uM), As 2 O 3  (60 uM), and doxorubicin (3 uM) exhibited 90% or higher inhibition of the HL60 AML cells. 
       FIG. 35  shows dose-response curves of Sensoril®, cytarabine, AS2O3, and doxorubicin on the HL60 lines, with the dose-response curve for Sensoril® showing an IC 50  for inhibition of HL60 cells of 67 ug/ml; the curve for cytarabine showing an IC 50  of 567 nM; the curve for As 2 O 3  showing an IC 50  of 2.3 uM; and the curve for doxorubicin showing an IC 50  of 79 nM. Combinations of Sensoril® with cytarabine, As2O3, and doxorubicin are expected to inhibit HL60 AML cells, for instance as shown in Table 71. 
     
       
         
           
               
             
               
                 TABLE 71 
               
             
            
               
                   
               
               
                 Potential dosing for further experiments on HL60 AML cells 
               
            
           
           
               
               
               
               
               
            
               
                   
                 Sensoril ® 
                 Cytarabine 
                 As2O3 
                 Doxorubicin 
               
               
                   
                 (ug/ml) 
                 (uM) 
                 (uM) 
                 (uM) 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 IC75 
                 109.65 
                 1.08 
                 3.01 
                 0.106 
               
               
                 IC50 
                 67.13 
                 0.567 
                 2.43 
                 0.079 
               
               
                 IC25 
                 41.10 
                 0.298 
                 1.72 
                 0.059 
               
               
                   
               
            
           
         
       
     
     Example VI 
     Inhibition of Triple Negative (TN) Breast Cancer PDCs (SB 30750) and TNBC Cell Line MDAMB-231 with AyuFlex®, Capros®, and Ayuric® 
     As shown for instance in  FIGS. 16-18 , AyuFlex®, Capros®, and Ayuric® showed exceptional anti-cancer effects against SB 36344 cells. AyuFlex® (10 ug/ml) was previously shown to inhibit TN breast cancer PDCs by 54%; Capros® (30 ug/ml), by 58%; and Ayuric® (30 ug/ml) by 47%. 
     In this Example, AyuFlex®, Capros®, and Ayuric® were each applied to different PDCs than previous tests, TN breast cancer PDCs SB 30750, and tested via the anti-proliferation assay described above. As shown in  FIG. 36 , each of AyuFlex®, Capros®, and Ayuric® was applied to the SB 30750 PDCs alone in concentrations of 100, 30, and 10 ug/ml, and combined as follows: AyuFlex® (30 ug/ml)+Capros® (30 ug/ml), AyuFlex® (30 ug/ml)+Ayuric® (30 ug/ml), AyuFlex® (10 ug/ml)+Capros® (30 ug/ml), AyuFlex® (10 ug/ml)+Ayuric® (30 ug/ml), and Capros® (30 ug/ml)+Ayuric® (30 ug/ml). Docetaxel (3 uM), 5-FU (3 uM), and doxorubicin (10 uM) controls are shown at the far right of the Figure. Incubation times were 72 hours. AyuFlex®, Capros®, and Ayuric® each showed a dose-dependent effect in the SB 30750 PDCs, and showed similar anti-cancer, inhibitory effects to the inhibition seen in the PDC cells of a different subject (SB 36344) tested in  FIGS. 16 and 18 . 
     Also, AyuFlex®, Capros®, and Ayuric® were tested in the anti-proliferative assay discussed above in the TN Breast Cancer cell line MDAMB-231. AyuFlex®, Capros®, and Ayuric® were incubated for 72 hours and 120 hours. The dose-response efficacy of AyuFlex®, Capros®, and Ayuric® after a 72-hour period of incubation was similar to that observed after a 120-hour period of incubation with the MDAMB-231 cells. Without being bound by theory, possibly this was due to saturation of cell growth at the 120-hour time-point. 
     Surprisingly, AyuFlex®, Capros®, and Ayuric® extracts showed better activity in both PDCs than in the MDAMB-231 cell line. The MDAMB-231 cell line was established from pleural effusion of a Caucasian female with a metastatic mammary adenocarcinoma 1, and has a high stem cell percentage. The TNBC PDCs used in earlier studies (shown in  FIGS. 16-18 ) were from Grade II and TNM-T2N1aMx tumor, and the PDCs of the present Example (SB 30750) are from a Grade I and TNM-T2N1aMx tumor, both likely lower grade and non-metastatic tumors compared with MDAMB-231 cells. Without being bound by theory, it may be that the extracts are more active in earlier stage tumors than metastatic ones. 
     In summary, with the results of this Example VI and previous studies described herein, AyuFlex®, Capros®, and Ayuric® extracts demonstrated reproducible anti-cancer effects in 3 independent experiments using two different TNVC cancer patient derived cells. 
     
       
         
           
               
             
               
                 TABLE 72 
               
             
            
               
                   
               
               
                 Plate Controls 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                 Standard 
                   
                   
               
               
                   
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 100% Proliferation 
                 74408 
                 9061 
                 1413 
                 0.63 
               
               
                   
                 (Cell Control) 
                   
                   
                   
                   
               
               
                   
                 0% Proliferation 
                 53 
                 6 
                   
                   
               
               
                   
                 (Media only) 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 73 
               
             
            
               
                   
               
               
                 Doxorubicin control 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 16406 
                 23949 
                 31038 
                 78 
                 68 
                 58 
                 68 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 74 
               
             
            
               
                   
               
               
                 Inhibition of TNBC PDCs (SB 30750) by AyuFlex ®, Capros ®, and Ayuric ® 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Combination 
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 Avg 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex 100 ug/ml 
                 23242 
                 33738 
                 33268 
                 69 
                 55 
                 55 
                 60 
                 8 
               
               
                 Ayuflex 30 ug/ml 
                 27053 
                 33894 
                 37482 
                 64 
                 54 
                 50 
                 56 
                 7 
               
               
                 Ayuflex 10 ug/ml 
                 45510 
                 63323 
                 62574 
                 39 
                 15 
                 16 
                 23 
                 14 
               
               
                 Capros 100 ug/ml 
                 25381 
                 34302 
                 35137 
                 66 
                 54 
                 53 
                 58 
                 7 
               
               
                 Capros 30 ug/ml 
                 54237 
                 63005 
                 68764 
                 27 
                 15 
                 8 
                 17 
                 10 
               
               
                 Capros 10 ug/ml 
                 50700 
                 70923 
                 69595 
                 32 
                 5 
                 6 
                 14 
                 15 
               
               
                 Ayuric 100 ug/ml 
                 25631 
                 37388 
                 44444 
                 66 
                 50 
                 40 
                 52 
                 13 
               
               
                 Ayuric 30 ug/ml 
                 30833 
                 42730 
                 58052 
                 59 
                 43 
                 22 
                 41 
                 18 
               
               
                 Ayuric 10 ug/ml 
                 45673 
                 59207 
                 64851 
                 39 
                 20 
                 13 
                 24 
                 13 
               
               
                 Ayuflex 30 ug/ml + 
                 18056 
                 22928 
                 25595 
                 76 
                 69 
                 66 
                 70 
                 5 
               
               
                 Capros 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex 30 ug/ml + 
                 14857 
                 17538 
                 19683 
                 80 
                 76 
                 74 
                 77 
                 3 
               
               
                 Ayuric 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex 10 ug/ml + 
                 25519 
                 33410 
                 39663 
                 66 
                 55 
                 47 
                 56 
                 10 
               
               
                 Capros 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex 10 ug/ml + 
                 22904 
                 30026 
                 31629 
                 69 
                 60 
                 58 
                 62 
                 6 
               
               
                 Ayuric 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Capros 30 ug/ml + 
                 20621 
                 29043 
                 29474 
                 72 
                 61 
                 60 
                 65 
                 7 
               
               
                 Ayuric 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Docetaxel @ 3 uM 
                 24001 
                 35688 
                 39462 
                 68 
                 52 
                 47 
                 56 
                 11 
               
               
                 5-FU @ 3 uM 
                 41465 
                 63866 
                 64261 
                 44 
                 14 
                 14 
                 24 
                 18 
               
               
                 Doxorubicin @ 10 uM 
                 16406 
                 23949 
                 31038 
                 78 
                 68 
                 58 
                 68 
                 10 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 75 
               
             
            
               
                   
               
               
                 Inhibition of MDAMB-231 cell line by AyuFlex ®, Capros ®, and 
               
               
                 Ayuric ® (72 hour incubation) 
               
            
           
           
               
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Combination 
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 Avg 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex 100 ug/ml 
                 58945 
                 77704 
                 76140 
                 44 
                 27 
                 28 
                 33 
                 10 
               
               
                 Ayuflex 30 ug/ml 
                 80393 
                 98268 
                 102334 
                 24 
                 7 
                 3 
                 11 
                 11 
               
               
                 Ayuflex 10 ug/ml 
                 92286 
                 122115 
                 122215 
                  13* 
                 −15 
                 −16 
                 −15 
                 0 
               
               
                 Capros 100 ug/ml 
                 70641 
                 92822 
                 95695 
                 33 
                 12 
                 10 
                 18 
                 13 
               
               
                 Capros 30 ug/ml 
                 108078 
                 130370 
                 132500 
                 −2 
                 −23 
                 −25 
                 −17 
                 13 
               
               
                 Capros 10 ug/ml 
                 97875 
                 128124 
                 134081 
                  7 
                 −21 
                 −27 
                 −13 
                 18 
               
               
                 Ayuric 100 ug/ml 
                 57505 
                 70158 
                 69844 
                 46 
                 34 
                 34 
                 38 
                 7 
               
               
                 Ayuric 30 ug/ml 
                 75642 
                 98513 
                 97248 
                 29 
                 7 
                 8 
                 15 
                 12 
               
               
                 Ayuric 10 ug/ml 
                 83078 
                 106876 
                 105518 
                 21 
                 −1 
                 0 
                 7 
                 13 
               
               
                 Ayuflex 30 ug/ml + 
                 74411 
                 92848 
                 90342 
                 30 
                 12 
                 15 
                 19 
                 9 
               
               
                 Capros 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex 30 ug/ml + 
                 70949 
                 87917 
                 86978 
                 33 
                 17 
                 18 
                 23 
                 9 
               
               
                 Ayuric 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex 10 ug/ml + 
                 85852 
                 108852 
                 101693 
                 19 
                 −3 
                 4 
                 7 
                 11 
               
               
                 Capros 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex 10 ug/ml + 
                 70288 
                 95788 
                 94340 
                 34 
                 9 
                 11 
                 18 
                 14 
               
               
                 Ayuric 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Capros 30 ug/ml + 
                 75562 
                 97654 
                 99317 
                 29 
                 8 
                 6 
                 14 
                 13 
               
               
                 Ayuric 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Docetaxel @ 3 uM 
                 35229 
                 52837 
                 50750 
                 67 
                 50 
                 52 
                 56 
                 9 
               
               
                 5-FU @ 3 uM 
                 83364 
                 111466 
                 110358 
                 21 
                 −5 
                 −4 
                 4 
                 15 
               
               
                 Doxorubicin 10 uM 
                 23120 
                 28746 
                 28430 
                 78 
                 73 
                 73 
                 75 
                 3 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 76 
               
             
            
               
                   
               
               
                 Inhibition of MDAMB-231 cell line by AyuFlex ®, Capros ®, and 
               
               
                 Ayuric ® (120 hour incubation) 
               
            
           
           
               
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Combination 
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 Avg 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Ayuflex 100 ug/ml 
                 56389 
                 77349 
                 72974 
                 56 
                 39 
                 43 
                 46 
                 9 
               
               
                 Ayuflex 30 ug/ml 
                 78537 
                 95588 
                 95641 
                 39 
                 25 
                 25 
                 30 
                 8 
               
               
                 Ayuflex 10 ug/ml 
                 95138 
                 130502 
                 131567 
                  25* 
                 −2 
                 −3 
                 −3 
                 1 
               
               
                 Capros 100 ug/ml 
                 70583 
                 95001 
                 98086 
                  45* 
                 26 
                 23 
                 24 
                 2 
               
               
                 Capros 30 ug/ml 
                 125560 
                 155502 
                 161033 
                  2* 
                 −22 
                 −26 
                 −24 
                 3 
               
               
                 Capros 10 ug/ml 
                 122241 
                 159839 
                 154609 
                  4* 
                 −25 
                 −21 
                 −23 
                 3 
               
               
                 Ayuric 100 ug/ml 
                 58558 
                 77747 
                 74476 
                 54 
                 39 
                 42 
                 45 
                 8 
               
               
                 Ayuric 30 ug/ml 
                 89163 
                 111142 
                 108375 
                 30 
                 13 
                 15 
                 19 
                 9 
               
               
                 Ayuric 10 ug/ml 
                 102684 
                 130666 
                 135088 
                  20* 
                 −2 
                 −6 
                 −4 
                 2 
               
               
                 Ayuflex 30 ug/ml + 
                 71269 
                 97666 
                 90166 
                 44 
                 24 
                 29 
                 32 
                 11 
               
               
                 Capros 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex 30 ug/ml + 
                 72855 
                 89938 
                 87457 
                 43 
                 30 
                 32 
                 35 
                 7 
               
               
                 Ayuric 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex 10 ug/ml + 
                 88342 
                 111648 
                 115247 
                  31* 
                 13 
                 10 
                 11 
                 2 
               
               
                 Capros 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Ayuflex 10 ug/ml + 
                 81695 
                 99895 
                 96079 
                 36 
                 22 
                 25 
                 28 
                 8 
               
               
                 Ayuric 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Capros 30 ug/ml + 
                 78312 
                 100528 
                 103458 
                 39 
                 21 
                 19 
                 26 
                 11 
               
               
                 Ayuric 30 ug/ml 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Docetaxel @ 3 uM 
                 28378 
                 36725 
                 30693 
                 78 
                 71 
                 76 
                 75 
                 3 
               
               
                 5-FU @ 3 uM 
                 102508 
                 140779 
                 134513 
                  20* 
                 −10 
                 −5 
                 −8 
                 3 
               
               
                 Doxorubicin 10 uM 
                 7939 
                 11844 
                 10217 
                 94 
                 91 
                 92 
                 92 
                 2 
               
               
                   
               
            
           
         
       
     
     Example VII 
     Inhibition of Small Cell Lung Cancer Cells with Different Extracts 
     SHP-77 (small cell lung cancer) cells were incubated for 72 hours with 100 ug/ml, 30 ug/ml, and 10 ug/ml each of AyuFlex®, Capros® Phyto-BGS®, PrimaVie®, Sensoril®, Ayuric®, and Crominex®, and tested according to the anti-proliferative assay described above. Cells were plated at 1.25 k/well in 384 well plates. Standard of care drug docetaxel was also tested at a 10 uM concentration as a control. Anti-proliferative assays were generally performed as discussed above, in triplicate, with detection reagent CellTiterGlo® as discussed for instance in Example I. Tables 77-79 show data for controls and that the data passed quality control parameters. 
     
       
         
           
               
             
               
                 TABLE 77 
               
             
            
               
                   
               
               
                 Plate Controls 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                 Standard 
                   
                   
               
               
                   
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 100% Proliferation 
                 49047 
                 3203 
                 30 
                 0.79 
               
               
                   
                 (Cell Control) 
                   
                   
                   
                   
               
               
                   
                 0% Proliferation 
                 1613 
                 79 
                   
                   
               
               
                   
                 (Media only) 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 78 
               
             
            
               
                   
               
               
                 Docetaxel control 
               
               
                 Docetaxel @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 6365 
                 5908 
                 6786 
                 90 
                 91 
                 89 
                 90(1) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 79 
               
             
            
               
                   
               
               
                 Doxorubicin control 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 4634 
                 5086 
                 5168 
                 94 
                 93 
                 93 
                 93(1) 
               
               
                   
               
            
           
         
       
     
     Docetaxel, a drug currently used for the treatment of small cell lung cancer, exhibited 90% inhibition of cell proliferation at the 10 uM concentration tested. The assay control, doxorubicin, exhibited 93% inhibition at the 10 uM concentration tested. 
     Of the extracts tested, AyuFlex® and Ayuric® exhibited the most potent anti-cancer effect on the SHP-77 small cell lung cancer cell line, exhibiting 98% inhibition at 100 ug/ml. The anti-cancer activity of the extracts on SHP-77 cells is similar to that of H-358 cells (non-small cell cancer, NSCLC, shown in Example III), with Ayuric® and AyuFlex® also showing the best anti-cancer effect in those cells. 
     Example VIII 
     Prostate cancer (PC-3) cells and Ovarian cancer (OVCAR-3) cells were incubated for 72 hours with 100 ug/ml, 30 ug/ml, and 10 ug/ml each of AyuFlex®, Capros® Phyto-BGS®, PrimaVie®, Sensoril®, Ayuric®, and Crominex®, and tested according to the anti-proliferative assay described above. Cells were plated at 1.25 k/well in 384 well plates. Standard of care drug docetaxel and control doxorubicin were each also tested at a 10 uM concentration. Anti-proliferative assays were generally performed as discussed in Example I above, in triplicate, with detection reagent CellTiterGlo. Tables 80-82 show data for controls for assays on prostate cancer (PC-3 cells), and that the data passed quality control parameters. Tables 83-85 show data for controls for assays on ovarian cancer (OVCAR-3 cells), and that the data passed quality control parameters. 
     
       
         
           
               
             
               
                 TABLE 80 
               
             
            
               
                   
               
               
                 Plate Controls 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                 Standard 
                   
                   
               
               
                   
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 100% Proliferation 
                 54196 
                 3632 
                 415 
                 0.80 
               
               
                   
                 (Cell Control) 
                   
                   
                   
                   
               
               
                   
                 0% Proliferation 
                 131 
                 37 
                   
                   
               
               
                   
                 (Media only) 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 81 
               
             
            
               
                   
               
               
                 Docetaxel control 
               
               
                 Docetaxel @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 25757 
                 18168 
                 24121 
                 53 
                 67 
                 56 
                 58(7) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 82 
               
             
            
               
                   
               
               
                 Doxorubicin control 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 16210 
                 16521 
                 15937 
                 70 
                 70 
                 71 
                 70(1) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 83 
               
             
            
               
                   
               
               
                 Plate Controls 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 Standard 
                   
                   
               
               
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 100% Proliferation 
                 45784 
                 1777 
                 595 
                 0.88 
               
               
                 (Cell Control) 
                   
                   
                   
                   
               
               
                 0% Proliferation 
                 77 
                 6 
                   
                   
               
               
                 (Media only) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 84 
               
             
            
               
                   
               
               
                 Docetaxel control 
               
               
                 Docetaxel @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 16072 
                 14601 
                 15965 
                 65 
                 68 
                 65 
                 66(2) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 85 
               
             
            
               
                   
               
               
                 Doxorubicin control 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 12071 
                 11517 
                 9891 
                 74 
                 75 
                 79 
                 76(2) 
               
               
                   
               
            
           
         
       
     
     With regard to prostate cancer (PC-3) cells, docetaxel, a drug currently used for the treatment of prostate cancer, exhibited 58% inhibition of cell proliferation at the 10 uM concentration tested. The assay control, doxorubicin, exhibited 70% inhibition at the 10 uM concentration tested. 
     Of the extracts tested, Ayuric® exhibited the most potent anti-cancer effect on the prostate cancer (PC-3) cell line, exhibiting 79% inhibition at 100 ug/ml; followed by AyuFlex®, which exhibited 50% inhibition in the assay at 100 ug/ml. 
     With regard to ovarian cancer (OVCAR-3) cells, docetaxel, a drug currently used for the treatment of ovarian cancer, exhibited 66% inhibition of cell proliferation at the 10 uM concentration tested. The assay control, doxorubicin, exhibited 76% inhibition at the 10 uM concentration tested. 
     Of the extracts tested, Ayuric® exhibited the most potent anti-cancer effect on the ovarian cancer cell line, exhibiting 92% inhibition of cell proliferation at 100 ug/ml, and 23% inhibition at 30 ug/ml. The next effective extracts against the proliferation of the OVCAR-3 ovarian cancer cells were AyuFlex® and Sensoril®, exhibiting 46% and 35% inhibition respectively at 100 ug/ml. 
     Example IX 
     Inhibition of AML (HL60 Cell Line) Cancer Cells with Hydroethanolic Extracts of  Withania somnifera  (Sensoril®-AWE) 
     Hydroethanolic extracts of  Withania somnifera  (Sensoril®-AWE) showed significant anti-cancer effects in AML cancer cells of the HL60 cell line. 2500 cells were plated per well in 384 well plates. Standard of care drug doxorubicin was included as a control at 10 uM. 6 different samples of Sensoril®-AWE, prepared with hydroethanolic extraction were tested in the anti-proliferative assay as generally described in Example I above. Extracts were prepared at 2 mg/ml concentrations in view of increased solubility and tested at 400 ug/ml, 2-fold serial dilutions, to prepare an 8-point Dose-Response Curve. CellTiter Glo was used as a detection reagent. Extracts were incubated with the AML HL60 cells for 72 hours. Table 86 provides information on hydroethanolic extract quantification and markers according to this invention. Full analysis of some Samples described below are as follows: Lot #WS09120: 19.34% w/w Withanolide glycosides, 7.52% w/w Withanolide Aglycones (as Withaferin A), 34.42% w/w Oligosaccharides; Lot #WS09820: 13.16% w/w Withanolide glycosides, 2.60% w/w Withanolide Aglycones (as Withaferin A), 26.52% w/w Oligosaccharides; Lot #WS09920: 12.68% w/w Withanolide glycosides, 2.71% w/w Withanolide Aglycones (as Withaferin A), 23.55% w/w Oligosaccharides; Lot #WS10020: 12.20% w/w Withanolide glycosides, 2.39% w/w Withanolide Aglycones (as Withaferin A), 24.64% w/w Oligosaccharides. Tables 87-90 show data for controls and that the data passed quality control parameters. 
     All hydroethanolic samples of  Withania somnifera  (Sensoril®-AWE) showed a sigmoidal saturated IC 50  dose-response curve with significant inhibition of AML HL60 cell proliferation. The most potent sample was the “Nepal” sample (Sample 1 of Table 91 below, Lot #WS09120), which showed an IC 50  of 17.5 ug/ml, with Sample 2, based solely on  Withania somnifera  leaves, following closely with an IC 50  of 19.2 ug/ml. Sample 3, 4, and 6 showed higher IC 50  values, ranging from 26-28 ug/ml, and Sample 5 showed lowest activity with an IC 50  of 48.7 ug/ml. See Table 91 for further information on Sample preparation and a listing of IC 50  s found for each Sample, Tables 92-97 for experimental data relating to the Samples, and  FIG. 43  for relevant dose-response curves. 
     
       
         
           
               
             
               
                 TABLE 86 
               
             
            
               
                   
               
               
                   Withania somnifera  (Ashwagandha) hydroethanolic extracts (Sensoril ®-AWE) of the present invention 
               
            
           
           
               
               
               
            
               
                   
                 Quantification using 
                   
               
               
                   
                 Withanoside-IV and 
                 Quantification using 
               
               
                   
                 Withanolide-A as Markers 
                 individual markers 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 Total 
                   
                 Total 
               
               
                 Sample name 
                 Class of 
                   
                 Content 
                 Content 
                 Content 
                 Content 
               
               
                 (Lot#) 
                 analytes 
                 Analytes 
                 (% w/w) 
                 (% w/w) 
                 (% w/w) 
                 (% w/w) 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 WS09420_Leaf 
                 Withanolide 
                 Withaferin A 
                 0.88 
                 1.45 
                 0.76 
                 1.57 
               
               
                 (70% ethanol) 
                 aglycones 
                 Withastromonolide 
                   
                   
                 0.10 
                   
               
               
                   
                   
                 Withanolide A 
                   
                   
                 0.05 
                   
               
               
                   
                   
                 Withanolide B 
                   
                   
                 0.02 
                   
               
               
                   
                   
                 27-Hydroxy Withanone 
                   
                   
                 0.08 
                   
               
               
                   
                   
                 Withanone 
                   
                   
                 0.00 
                   
               
               
                   
                 Withanolide 
                 Withanoside IV 
                 0.57 
                   
                 0.40 
                   
               
               
                   
                 glycosides 
                 Withanoside V 
                   
                   
                 0.17 
                   
               
               
                 WS09320_Root 
                 Withanolide 
                 Withaferin A 
                 1.03 
                 1.58 
                 0.99 
                 1.85 
               
               
                 (70% ethanol) 
                 aglycones 
                 Withastromonolide 
                   
                   
                 0.10 
                   
               
               
                   
                   
                 Withanolide A 
                   
                   
                 0.07 
                   
               
               
                   
                   
                 Withanolide B 
                   
                   
                 0.02 
                   
               
               
                   
                   
                 27-Hydroxy Withanone 
                   
                   
                 0.07 
                   
               
               
                   
                   
                 Withanone 
                   
                   
                 0.00 
                   
               
               
                   
                 Withanolide 
                 Withanoside IV 
                 0.55 
                   
                 0.40 
                   
               
               
                   
                 glycosides 
                 Withanoside V 
                   
                   
                 0.19 
                   
               
               
                 WS09220_Leaf:Root 
                 Withanolide 
                 Withaferin A 
                 0.71 
                 1.21 
                 0.56 
                 1.45 
               
               
                 (70% ethanol) 
                 aglycones 
                 Withastromonolide 
                   
                   
                 0.12 
                   
               
               
                   
                   
                 Withanolide A 
                   
                   
                 0.08 
                   
               
               
                   
                   
                 Withanolide B 
                   
                   
                 0.02 
                   
               
               
                   
                   
                 27-Hydroxy Withanone 
                   
                   
                 0.05 
                   
               
               
                   
                   
                 Withanone 
                   
                   
                 0.00 
                   
               
               
                   
                 Withanolide 
                 Withanoside IV 
                 0.49 
                   
                 0.43 
                   
               
               
                   
                 glycosides 
                 Withanoside V 
                   
                   
                 0.19 
                   
               
               
                 WS08920_Leaf 
                 Withanolide 
                 Withaferin A 
                 0.94 
                 1.45 
                 1.36 
                 2.11 
               
               
                 (100% ethanol) 
                 aglycones 
                 Withastromonolide 
                   
                   
                 0.07 
                   
               
               
                 Followed by water 
                   
                 Withanolide A 
                   
                   
                 0.02 
                   
               
               
                   
                   
                 Withanolide B 
                   
                   
                 0.00 
                   
               
               
                   
                   
                 27-Hydroxy Withanone 
                   
                   
                 0.12 
                   
               
               
                   
                   
                 Withanone 
                   
                   
                 0.00 
                   
               
               
                   
                 Withanolide 
                 Withanoside IV 
                 0.51 
                   
                 0.39 
                   
               
               
                   
                 glycosides 
                 Withanoside V 
                   
                   
                 0.16 
                   
               
               
                 WS09020_Root 
                 Withanolide 
                 Withaferin A 
                 0.64 
                 0.99 
                 0.60 
                 1.09 
               
               
                 (100% ethanol) 
                 aglycones 
                 Withastromonolide 
                   
                   
                 0.05 
                   
               
               
                 Followed by water 
                   
                 Withanolide A 
                   
                   
                 0.05 
                   
               
               
                   
                   
                 Withanolide B 
                   
                   
                 0.01 
                   
               
               
                   
                   
                 27-Hydroxy Withanone 
                   
                   
                 0.05 
                   
               
               
                   
                   
                 Withanone 
                   
                   
                 0.00 
                   
               
               
                   
                 Withanolide 
                 Withanoside IV 
                 0.34 
                   
                 0.23 
                   
               
               
                   
                 glycosides 
                 Withanoside V 
                   
                   
                 0.10 
                   
               
               
                 WS09120_Leaf:Root 
                 Withanolide 
                 Withaferin A 
                 1.75 
                 2.46 
                 1.64 
                 2.81 
               
               
                 (100% ethanol) 
                 aglycones 
                 Withastromonolide 
                   
                   
                 0.15 
                   
               
               
                   
                   
                 Withanolide A 
                   
                   
                 0.14 
                   
               
               
                   
                   
                 Withanolide B 
                   
                   
                 0.03 
                   
               
               
                   
                   
                 27-Hydroxy Withanone 
                   
                   
                 0.10 
                   
               
               
                   
                   
                 Withanone 
                   
                   
                 0.00 
                   
               
               
                   
                 Withanolide 
                 Withanoside IV 
                 0.70 
                   
                 0.47 
                   
               
               
                   
                 glycosides 
                 Withanoside V 
                   
                   
                 0.27 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 87 
               
             
            
               
                   
               
               
                 Assay Plate 1 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                 Standard 
                   
                   
               
               
                   
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 100% Proliferation 
                 21647 
                 1706 
                 2405 
                 0.76 
               
               
                   
                 (Cell Control) 
                   
                   
                   
                   
               
               
                   
                 0% Proliferation 
                 9 
                 1 
                   
                   
               
               
                   
                 (Media only) 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 88 
               
             
            
               
                   
               
               
                 Plate 1 - Doxorubicin 10 uM 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 17 
                 10 
                 13 
                 100 
                 100 
                 100 
                 0.0 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 89 
               
             
            
               
                   
               
               
                 Assay Plate 2 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                 Standard 
                   
                   
               
               
                   
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 100% Proliferation 
                 22538 
                 2131 
                 1439 
                 0.72 
               
               
                   
                 (Cell Control) 
                   
                   
                   
                   
               
               
                   
                 0% Proliferation 
                 16 
                 4 
                   
                   
               
               
                   
                 (Media only) 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 90 
               
             
            
               
                   
               
               
                 Plate 2 - Doxorubicin 10 uM 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 10 
                 11 
                 10 
                 100 
                 100 
                 100 
                 0.0 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 91 
               
             
            
               
                   
               
               
                 IC 50  of Different Examples of  Withania somnifera  (Sensoril ®-AWE)  
               
               
                 Hydroethanolic Extracts 
               
            
           
           
               
               
               
               
            
               
                   
                 
                   Withania somnifera 
                 
                 IC 50   
                   
               
               
                 Sample/Lot # 
                 part used 
                 (ug/ml) 
                 Extraction Process 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 1/#WS09120 
                 Nepal (Root + Leaf) 
                 17.5 
                 Ethanol followed by aqueous 
               
               
                 2/#WS08920 
                 Leaf 
                 19.2 
                 Ethanol followed by aqueous 
               
               
                 3/#WS09920 
                 Root 
                 28.1 
                 Ethanol followed by aqueous 
               
               
                 4/#WS09820 
                 Leaf + Root 
                 26.7 
                 Ethanol followed by aqueous 
               
               
                 5/#WS09020 
                 Leaf + Root 
                 48.7 
                 Ethanol followed by aqueous 
               
               
                 6/#WS10020 
                 Leaf + Root 
                 27.78 
                 Ethanol followed by aqueous 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 92 
               
             
            
               
                   
               
               
                   Withania somnifera  (Sensoril ®-AWE) Hydroethanolic 
               
               
                 Extract Sample 1 (“Nepal,” Root + Leaf) 
               
               
                 Sensoril (Nepal) 
               
            
           
           
               
               
               
               
            
               
                   
                 Conc. 
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 (ug · ml 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 1 
                 400 
                 29 
                 24 
                 100 
                 100 
                 100 
               
               
                 2 
                 200 
                 19 
                 18 
                 100 
                 100 
                 100 
               
               
                 3 
                 100 
                 35 
                 29 
                 100 
                 100 
                 100 
               
               
                 4 
                 50 
                 100 
                 92 
                 100 
                 100 
                 100 
               
               
                 5 
                 25 
                 1824 
                 1972 
                 92 
                 91 
                 91 
               
               
                 6 
                 13 
                 21358 
                 21524 
                 1 
                 1 
                 1 
               
               
                 7 
                 6 
                 26242 
                 25684 
                 −21 
                 −19 
                 −20 
               
               
                 8 
                 3 
                 23562 
                 21011 
                 −9 
                 3 
                 −3 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 93 
               
             
            
               
                   
               
               
                   Withania somnifera  (Sensoril ®-AWE) Hydroethanolic Extract Sample 2 
               
               
                 Sensoril Lot # WS08920 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 Conc. 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 1 
                 400 
                 30 
                 31 
                 100 
                 100 
                 100 
               
               
                 2 
                 200 
                 18 
                 17 
                 100 
                 100 
                 100 
               
               
                 3 
                 100 
                 69 
                 64 
                 100 
                 100 
                 100 
               
               
                 4 
                 50 
                 272 
                 251 
                 99 
                 99 
                 99 
               
               
                 5 
                 25 
                 6670 
                 5389 
                 69 
                 75 
                 72 
               
               
                 6 
                 13 
                 23535 
                 24001 
                 −9 
                 −11 
                 −10 
               
               
                 7 
                 6 
                 28535 
                 27853 
                 −32 
                 −29 
                 −30 
               
               
                 8 
                 3 
                 26615 
                 27234 
                 −23 
                 −26 
                 −24 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 94 
               
             
            
               
                   
               
               
                   Withania somnifera  (Sensoril ®-AWE) Hydroethanolic Extract Sample 3 
               
               
                 Sensoril # WS 9920 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 Conc. 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 1 
                 400 
                 12 
                 9 
                 100 
                 100 
                 100 
               
               
                 2 
                 200 
                 23 
                 25 
                 100 
                 100 
                 100 
               
               
                 3 
                 100 
                 84 
                 66 
                 100 
                 100 
                 100 
               
               
                 4 
                 50 
                 806 
                 636 
                 96 
                 97 
                 97 
               
               
                 5 
                 25 
                 19036 
                 16768 
                 12 
                 23 
                 17 
               
               
                 6 
                 13 
                 27425 
                 24977 
                 −27 
                 −15 
                 −21 
               
               
                 7 
                 6 
                 26766 
                 26654 
                 −24 
                 −23 
                 −23 
               
               
                 8 
                 3 
                 27133 
                 26806 
                 −25 
                 −24 
                 −25 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 95 
               
             
            
               
                   
               
               
                   Withania somnifera  (Sensoril ®-AWE) Hydroethanolic Extract Sample 4 
               
               
                 Sensoril # WS 9820 
               
            
           
           
               
               
               
               
            
               
                   
                 Conc. 
                 Raw Data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 (ug, ml 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 1 
                 400 
                 23 
                 21 
                 100 
                 100 
                 100 
               
               
                 2 
                 200 
                 74 
                 30 
                 100 
                 100 
                 100 
               
               
                 3 
                 100 
                 112 
                 87 
                 100 
                 100 
                 100 
               
               
                 4 
                 50 
                 745 
                 620 
                 97 
                 97 
                 97 
               
               
                 5 
                 25 
                 15188 
                 15170 
                 30 
                 30 
                 30 
               
               
                 6 
                 13 
                 24973 
                 24967 
                 −15 
                 −15 
                 −15 
               
               
                 7 
                 6 
                 26116 
                 24970 
                 −21 
                 −15 
                 −18 
               
               
                 8 
                 3 
                 25775 
                 25723 
                 −19 
                 −19 
                 −19 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 96 
               
             
            
               
                   
               
               
                   Withania somnifera  (Sensoril ®-AWE) Hydroethanolic Extract Sample 5 
               
               
                 Sensoril # WS 9020 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 Conc. 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 1 
                 400 
                 44 
                 85 
                 100 
                 100 
                 100 
               
               
                 2 
                 200 
                 88 
                 68 
                 100 
                 100 
                 100 
               
               
                 3 
                 100 
                 393 
                 357 
                 98 
                 98 
                 98 
               
               
                 4 
                 50 
                 10464 
                 9713 
                 52 
                 55 
                 53 
               
               
                 5 
                 25 
                 21450 
                 20206 
                 1 
                 7 
                 4 
               
               
                 6 
                 13 
                 24551 
                 22741 
                 −13 
                 −5 
                 −9 
               
               
                 7 
                 6 
                 21259 
                 19835 
                 2 
                 8 
                 5 
               
               
                 8 
                 3 
                 21984 
                 19733 
                 −2 
                 9 
                 4 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 97 
               
             
            
               
                   
               
               
                   Withania somnifera  (Sensoril ®-AWE) Hydroethanolic Extract Sample 6 
               
               
                 Sensoril # WS 10020 
               
            
           
           
               
               
               
            
               
                   
                 Raw data 
                 % Inhibition 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Doses 
                 Conc. 
                 n1 
                 n2 
                 n1 
                 n2 
                 Avg 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 1 
                 400 
                 29 
                 27 
                 100 
                 100 
                 100 
               
               
                 2 
                 200 
                 25 
                 22 
                 100 
                 100 
                 100 
               
               
                 3 
                 100 
                 78 
                 66 
                 100 
                 100 
                 100 
               
               
                 4 
                 50 
                 483 
                 356 
                 98 
                 98 
                 98 
               
               
                 5 
                 25 
                 15121 
                 12884 
                 30 
                 40 
                 35 
               
               
                 6 
                 13 
                 23885 
                 21962 
                 −10 
                 −1 
                 −6 
               
               
                 7 
                 6 
                 21871 
                 21417 
                 −1 
                 1 
                 0 
               
               
                 8 
                 3 
                 21346 
                 20448 
                 1 
                 6 
                 3 
               
               
                   
               
            
           
         
       
     
     Example X 
     Inhibition of Histiocytic Lymphoma and Pancreatic Cancer Cells with Different Extracts 
     Histiocytic lymphoma (U-937) cells and Pancreatic cancer (Panc-1) cells were incubated for 72 hours with 100 ug/ml, 30 ug/ml, and 10 ug/ml each of fresh samples of AyuFlex®, Capros®, Phyto-BGS®, 2 different samples of Shilajit (PrimaVie®) extracts, several different samples of hydroethanolic  Withania somnifera  (Sensoril®-AWE) extracts, and Ayuric®. Cells were plated at 1.25 k/well in 384 well plates. Standard of care drug doxorubicin was also tested at a 10 uM concentration. Anti-proliferative assays were generally performed as described in Example I above, in triplicate, with detection reagent CellTiterGlo. Tables 98-99 show data for controls for assays on histiocytic lymphoma (U-937) cells, and that the data passed quality control parameters. Tables 101-102 show data for controls for assays on pancreatic cancer (Panc-1) cells, and that the data passed quality control parameters. 
     Tables 100 and 103 show data relating to the inhibition of histiocytic lymphoma (U-937) cells and pancreatic cancer (Panc-1) cells by extracts of this invention. The assay is generally as described in Example I. 
     
       
         
           
               
             
               
                 TABLE 98 
               
             
            
               
                   
               
               
                 Assay Plate 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                 Standard 
                   
                   
               
               
                   
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 100% Proliferation 
                 52629 
                 4256 
                 1847 
                 0.76 
               
               
                   
                 (Cell Control) 
                   
                   
                   
                   
               
               
                   
                 0% Proliferation 
                 29 
                 4 
                   
                   
               
               
                   
                 (Media only) 
                   
                   
                   
                   
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 99 
               
             
            
               
                   
               
               
                 Doxorubicin 10 uM 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 58 
                 50 
                 70 
                 100 
                 100 
                 100 
                 100(0) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 100 
               
             
            
               
                   
               
               
                 Inhibition of histiocytic lymphoma cancer cell (U-937 cell line) proliferation with different extracts 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 S. 
                   
                 % Inhibition 
                 Avg % 
                   
                 Avg % 
                   
                 Avg % 
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 No. 
                 Extracts 
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
                 Inhibition 
                 SD 
                 Inhibition 
                 SD 
                 Inhibition 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                 Ayuflex 
                 61 
                 58 
                 64 
                 12 
                 7 
                 10 
                 −9 
                 −11 
                 0 
                 61 
                 3 
                 9 
                 3 
                 −6 
                 6 
               
               
                 2 
                 Capros 
                 −6 
                 −7 
                 −8 
                 −12 
                 −6 
                 2 
                 −6 
                 −6 
                 −3 
                 −7 
                 1 
                 −6 
                 7 
                 −5 
                 2 
               
               
                 3 
                 Phyto-BGS 
                 −19 
                 −19 
                 −17 
                 −11 
                 −9 
                 −3 
                 −9 
                 −8 
                 −3 
                 −18 
                 1 
                 −8 
                 4 
                 −7 
                 3 
               
               
                 4 
                 Primavie 
                 −3 
                 −13 
                 −1 
                 −13 
                 −13 
                 −15 
                 −7 
                 −12 
                 −18 
                 −5 
                 6 
                 −13 
                 1 
                 −12 
                 6 
               
               
                 5 
                 Sensoril 
                 −19 
                 −5 
                 0 
                 4 
                 10 
                 22 
                 11 
                 9 
                 −3 
                 −8 
                 10 
                 12 
                 9 
                 6 
                 8 
               
               
                 6 
                 Ayuric 
                 44 
                 45 
                 45 
                 −4 
                 1 
                 13 
                 −8 
                 −11 
                 −12 
                 45 
                 0 
                 4 
                 9 
                 −10 
                 2 
               
               
                 7 
                 Sensoril 
                 99 
                 99 
                 99 
                 97 
                 97 
                 98 
                 42 
                 40 
                 38 
                 99 
                 0 
                 97 
                 0 
                 40 
                 2 
               
               
                   
                 (Nepal) 
               
               
                 8 
                 Sensoril 
                 6 
                 3 
                 8 
                 21 
                 20 
                 20 
                 20 
                 21 
                 21 
                 6 
                 2 
                 20 
                 0 
                 21 
                 0 
               
               
                   
                 (Patel) 
               
               
                 9 
                 Sensoril 
                 9 
                 6 
                 8 
                 −4 
                 4 
                 8 
                 1 
                 −8 
                 −1 
                 7 
                 1 
                 2 
                 6 
                 −3 
                 5 
               
               
                   
                 (Fitochem) 
               
               
                 10 
                 Primavie 
                 30 
                 29 
                 36 
                 16 
                 18 
                 15 
                 NA 
                 NA 
                 NA 
                 31 
                 4 
                 16 
                 2 
                 NA 
                 NA 
               
               
                   
                 (Fitochem) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 101 
               
             
            
               
                   
               
               
                 Assay Plate 
               
               
                 Analysis 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 Standard 
                   
                   
               
               
                   
                 Average 
                 Deviation 
                 Assay fold 
                 Z′ 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 100% Proliferation 
                 63086 
                 3225 
                 1043 
                 0.85 
               
               
                 (Cell Control) 
                   
                   
                   
                   
               
               
                 0% Proliferation 
                 61 
                 9 
                   
                   
               
               
                 (Media only) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 102 
               
             
            
               
                   
               
               
                 Doxorubicin 10 uM 
               
               
                 Doxorubicin @ 10 uM 
               
            
           
           
               
               
               
            
               
                   
                 % Inhibition 
                   
               
            
           
           
               
               
               
            
               
                 Average 
                   
                 Average 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 n1 
                 n2 
                 n3 
                 n1 
                 n2 
                 n3 
                 (SD) 
               
               
                   
               
               
                 3908 
                 4011 
                 3437 
                 94 
                 94 
                 95 
                 94(0) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 103 
               
             
            
               
                   
               
               
                 Inhibition of pancreatic cancer cell (Panc-1 cell line) proliferation with different extracts 
               
            
           
           
               
               
               
               
            
               
                   
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 S. 
                   
                 % Inhibition 
                 Avg % 
                   
                 Avg % 
                   
                 Avg % 
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 No. 
                 Extracts 
                 100 ug/ml 
                 30 ug/ml 
                 10 ug/ml 
                 Inhibition 
                 SD 
                 Inhibition 
                 SD 
                 Inhibition 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                 Ayuflex 
                 36 
                 36 
                 35 
                 2 
                 −1 
                 −1 
                 −4 
                 0 
                 4 
                 36 
                 1 
                 0 
                 2 
                 0 
                 4 
               
               
                 2 
                 Capros 
                 15 
                 9 
                 12 
                 0 
                 4 
                 1 
                 10 
                 12 
                 11 
                 12 
                 3 
                 2 
                 2 
                 11 
                 1 
               
               
                 3 
                 Phyto-BGS 
                 6 
                 5 
                 0 
                 3 
                 4 
                 4 
                 5 
                 5 
                 4 
                 4 
                 3 
                 4 
                 1 
                 5 
                 1 
               
               
                 4 
                 Primavie 
                 −1 
                 3 
                 5 
                 6 
                 7 
                 4 
                 1 
                 8 
                 2 
                 2 
                 3 
                 6 
                 1 
                 4 
                 4 
               
               
                 5 
                 Sensoril 
                 13 
                 8 
                 9 
                 9 
                 9 
                 5 
                 8 
                 18 
                 15 
                 10 
                 3 
                 8 
                 2 
                 14 
                 5 
               
               
                   
                 (Sakti) 
               
               
                 6 
                 Ayuric 
                 32 
                 29 
                 35 
                 12 
                 14 
                 13 
                 18 
                 18 
                 22 
                 32 
                 3 
                 13 
                 1 
                 19 
                 2 
               
               
                 7 
                 Sensoril 
                 55 
                 56 
                 55 
                 71 
                 67 
                 60 
                 24 
                 30 
                 25 
                 55 
                 1 
                 66 
                 6 
                 26 
                 3 
               
               
                   
                 (Nepal) 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 8 
                 Sensoril 
                 22 
                 22 
                 28 
                 Not Done 
                 27 
                 23 
                 25 
                 24 
                 4 
                 NA 
                 NA 
                 25 
                 2 
               
               
                   
                 Patel 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 9 
                 Sensoril 
                 17 
                 16 
                 21 
                 20 
                 18 
                 14 
                 Not Done 
                 18 
                 3 
                 18 
                 3 
                 NA 
                 NA 
               
               
                   
                 (Fitochem) 
               
               
                   
               
            
           
         
       
     
       FIG. 44  shows the anti-cancer, anti-proliferative effects of extract compositions of this invention on histiocytic lymphoma (U-937 cell line) cancer cells. Hydroethanolic extracts of  Withania somnifera  (Sensoril®-AWE) showed the most potent anti-cancer effects against the histiocytic lymphoma cells, with about 100% inhibition at 100 ug/ml and 30 ug/ml doses, and 40% inhibition at the 10 ug/ml dose. The next most effective extracts were Ayuflex® and Ayuric®, which showed 60% and 45% inhibition at the 100 ug/ml doses. The assay control, doxorubicin, showed 100% inhibition at 10 uM. 
       FIG. 45  shows the anti-cancer, anti-proliferative effects of extract compositions of this invention on pancreatic cancer (Panc-1 cell line) cancer cells. Hydroethanolic extracts of  Withania somnifera  (Sensoril®-AWE) showed the most potent anti-cancer effect, with 50-60% at 100 ug/ml and 30 ug/ml doses, followed by 26% inhibition at the 10 ug/ml concentration. The next most effective extract was Ayuflex®, showing greater than 30% inhibition of pancreatic cancer cell growth (Panc-1 cell line) at 100 ug/ml. The assay control, doxorubicin, showed 94% inhibition at 10 uM. 
     Discussion 
     Compositions of the present invention provide anti-cancer activity, as shown by the remarkable inhibition of cancer cell proliferation by compositions of this invention in the above Examples. Anti-proliferative activity was seen with the present compositions in glioma cancer cells, breast cancer cells, chronic lymphocytic leukemia cancer cells, acute myeloid leukemia cancer cells, small cell lung cancer cells, non-small lung cancer cells, colon cancer cells, prostate cancer cells, pancreatic cancer cells, and ovarian cancer cells. Inhibition was seen with different extracts on difference cancer cell types, sometimes with similar or even synergistic effects with combinations of extracts and/or extract(s) with a known anti-cancer drug. See for instance  FIGS. 8, 11, 15, 18, 27-28, 31-33, and 36 . 
     For instance, Ayuflex® and Ayuric® inhibited the proliferation of several cancer cell types, including pancreatic, ovarian, prostatic, small cell lung cancer, and colon cancer cells. Ayuflex® and Ayuric® may be administered for instance with standard non-small cell lung cancer and colon cancer drugs, and other drugs useful in treating cancer, including a low dose of said drugs to achieve optimal efficacy and lower side effects of the drugs. In an embodiment, a composition of the present invention may be used with a standard of care cancer drug to minimize the amount of standard of care drug given to a subject. Given the harsh adverse and toxic effects of many known cancer drugs, the present invention provides a useful method for co-administration with such drugs. 
     Also for instance Sensoril® showed potent anti-cancer and anti-proliferative activity against different subtypes of AML, alone and synergistically in combination with drugs relevant to AML (Acute Myeloid Leukemia) and subtype APL (Acute Promyelocytic Leukemia (APL, APML)) treatment such as arsenic trioxide and doxorubicin. Hydroethanolic Sensoril®-AWE compositions were shown to be effective against AML HL60 cell line cancer cells. Sensoril® or Hydroethanolic Sensoril® (Sensoril®-AWE) may be administered with standard AML drugs, including at a low dose of the AML drugs, to achieve optimal efficacy and lower side effects of the drugs. 
     Several embodiments of the present invention are set out herein. In addition to the below, embodiments supported by the above Examples are intended as general embodiments of the invention, similar to the below. 
     A method of inhibiting the proliferation of cancer cells and/or cancer-associated cells comprising the steps of providing a composition comprising an extract of  Terminalia chebula  fruits, and applying said composition to said cancer cells to inhibit proliferation of the cells. In an embodiment, said composition is a standardized aqueous extract of the  Terminalia chebula  fruits, and said extract is Ayuflex®. In an embodiment, said cancer cells are glioma cells, breast cancer cells, chronic lymphocytic leukemia cells, acute myeloid leukemia cells, non-small cell lung cancer cells, small cell lung cancer cells, prostate cancer cells, ovarian cancer cells, pancreatic cancer cells, and/or colon cancer cells. In an embodiment, said cancer cells are chronic lymphocytic leukemia cells, glioma cells, prostate cancer cells, ovarian cancer cells, pancreatic cancer cells, breast cancer “HR+” cells, breast cancer “Her2+” cells, or breast cancer “triple negative” cells. A method of treating cancer in a subject in need thereof, and/or enhancing the treatment of cancer in a subject in need thereof, comprising the steps of providing a composition comprising an extract of  Terminalia chebula  fruits, and administering an effective amount of said composition to treat and/or enhance treatment of cancer in the subject. In an embodiment, said composition is a standardized aqueous extract of the  Terminalia chebula  fruits, and said extract is Ayuflex®. In an embodiment, said cancer is glioma, breast cancer, chronic lymphocytic leukemia, acute myeloid leukemia, non-small cell lung cancer, prostate cancer cells, ovarian cancer cells, pancreatic cancer cells, and/or colon cancer. In an embodiment, said cancer is chronic lymphocytic leukemia, glioma, prostate cancer cells, ovarian cancer cells, pancreatic cancer cells, breast cancer “HR+”, breast cancer “Her2+”, or breast cancer “triple negative”. 
     A method of inhibiting the proliferation of cancer cells comprising the steps of providing a composition comprising an extract of  Terminalia bellerica  fruits, and applying said composition to said cancer cells to inhibit proliferation of the cells. In an embodiment, said composition is a standardized aqueous extract of the  Terminalia bellerica  fruits, and said extract is Ayuric®. In an embodiment, said cancer cells are glioma cells, breast cancer cells, chronic lymphocytic leukemia cells, acute myeloid leukemia cells, non-small cell lung cancer cells, prostate cancer cells, ovarian cancer cells, pancreatic cancer cells, and/or colon cancer cells. In an embodiment, said cancer cells are chronic lymphocytic leukemia cells, breast cancer “HR+” cells, breast cancer “Her2+” cells, breast cancer “triple negative” cells, acute myeloid leukemia cells, non-small cell lung cancer cells, prostate cancer cells, ovarian cancer cells, pancreatic cancer cells, and/or colon cancer cells. A method of treating cancer in a subject in need thereof, and/or enhancing the treatment of cancer in a subject in need thereof, comprising the steps of providing a composition comprising an extract of  Terminalia bellerica  fruits, and administering said composition in an effective amount to treat and/or enhance treatment of cancer in the subject. In an embodiment, said composition is a standardized aqueous extract of  Terminalia bellerica  fruits, and said extract is Ayuric®. In an embodiment, said cancer is glioma, breast cancer, chronic lymphocytic leukemia, acute myeloid leukemia, non-small cell lung cancer, prostate cancer cells, ovarian cancer cells, pancreatic cancer cells, and/or colon cancer. In an embodiment, said cancer is chronic lymphocytic leukemia, breast cancer “HR+”, breast cancer “Her2+”, breast cancer “triple negative”, or acute myeloid leukemia. 
     A method of inhibiting the proliferation of cancer cells comprising the steps of providing a composition comprising an extract of  Phyllanthus emblica  fruits, and applying said composition to said cancer cells to inhibit proliferation of the cells. The method of claim  17 , wherein said composition is a standardized aqueous extract of  Phyllanthus emblica  fruits, and said extract is Capros®. In an embodiment, said cancer cells are glioma cells, breast cancer cells, chronic lymphocytic leukemia cells, and/or acute myeloid leukemia cells, non-small cell lung cancer cells, prostate cancer cells, ovarian cancer cells, pancreatic cancer cells, and/or colon cancer cells. In an embodiment, said cancer cells are chronic lymphocytic leukemia cells, breast cancer “Her2+” cells, or breast cancer “triple negative” cells. A method of treating cancer in a subject in need thereof and/or enhancing the treatment of cancer in a subject in need thereof, comprising the steps of providing a composition comprising an extract of  Phyllanthus emblica  fruits, and administering said composition in an effective amount to treat and/or enhance treatment of cancer in the subject. In an embodiment, said composition is a standardized aqueous extract of said  Phyllanthus emblica  fruits, and said extract is Capros®. In an embodiment, said cancer is glioma, breast cancer, chronic lymphocytic leukemia, acute myeloid leukemia, non-small cell lung cancer, prostate cancer cells, ovarian cancer cells, pancreatic cancer cells, and/or colon cancer. In an embodiment, said cancer is chronic lymphocytic leukemia, breast cancer “Her2+”, or breast cancer “triple negative.” 
     A method of inhibiting the proliferation of cancer cells comprising the steps of providing a composition comprising an extract of  Withania somnifera  leaves, roots, or roots plus leaves, and applying said composition to said cancer cells to inhibit proliferation of the cells. In an embodiment, said composition is a standardized aqueous extract or is a standardized hydroalcoholic extract of said leaves, roots, or roots plus leaves, and said extract is Sensoril®. In an embodiment, said cancer cells are glioma cells, breast cancer cells, chronic lymphocytic leukemia cells, acute myeloid leukemia cells including APL cells, non-small cell lung cancer cells, prostate cancer cells, ovarian cancer cells, pancreatic cancer cells, and/or colon cancer cells. In an embodiment, said cancer cells are acute myeloid leukemia cells or breast cancer “triple negative” cells. A method of treating cancer in a subject in need thereof, and/or enhancing the treatment of cancer in a subject in need thereof, comprising the steps of providing a composition comprising an extract of  Withania somnifera  leaves, roots, or roots plus leaves, and administering said composition in an effective amount to treat and/or enhance treatment of cancer in the subject. In an embodiment, said composition is a standardized aqueous extract or a standardized hydroalcoholic extract of the leaves, roots, or roots plus leaves, and said extract is Sensoril® or Sensoril®-AWE. In an embodiment, said cancer is glioma, breast cancer, chronic lymphocytic leukemia, acute myeloid leukemia including APL, non-small cell lung cancer, prostate cancer, ovarian cancer, pancreatic cancer, and/or colon cancer. In an embodiment, said cancer is acute myeloid leukemia or breast cancer “triple negative”. 
     A method of inhibiting the proliferation of cancer cells comprising the steps of providing a composition comprising an extract of Shilajit and applying said composition to said cancer cells to inhibit proliferation of the cells. In an embodiment, said extract is a standardized aqueous extract of Shilajit, and said extract is Primavie®. In an embodiment, said cancer cells are glioma cells, breast cancer cells, chronic lymphocytic leukemia cells, acute myeloid leukemia cells, non-small cell lung cancer cells, and/or colon cancer cells. In an embodiment, said cancer cells are acute myeloid leukemia cells. A method of treating cancer in a subject in need thereof, and/or enhancing the treatment of cancer in a subject in need thereof, comprising the steps of providing a composition comprising an extract of Shilajit, and administering said composition in an effective amount to treat and/or enhance treatment of cancer in the subject. In an embodiment, said extract is a standardized aqueous extract of Shilajit, and said extract is Primavie®. In an embodiment, said cancer is glioma, breast cancer, chronic lymphocytic leukemia, acute myeloid leukemia, non-small cell lung cancer, and/or colon cancer. In an embodiment, said cancer is acute myeloid leukemia. 
     A method of inhibiting the proliferation of cancer cells comprising the steps of providing a composition comprising an extract of  Azadirachta indica  leaves and twigs, and applying said composition to said cancer cells to inhibit proliferation of the cells. In an embodiment, said composition is a standardized aqueous extract, and said extract is PhytoBGS®. In an embodiment, said cancer cells are glioma cells, breast cancer cells, chronic lymphocytic leukemia cells, acute myeloid leukemia cells, non-small cell lung cancer cells, and/or colon cancer cells. In an embodiment, said cancer cells are acute myeloid leukemia cells or breast cancer “Her2+” cells. A method of treating cancer in a subject in need thereof, and/or enhancing the treatment of cancer in a subject in need thereof, comprising the steps of providing a composition comprising an extract of  Azadirachta indica  leaves and twigs, and administering said composition in an effective amount to treat and/or enhance treatment of cancer in the subject. In an embodiment, said composition is a standardized aqueous extract, and said extract is PhytoBGS®. In an embodiment, said cancer is glioma, breast cancer, chronic lymphocytic leukemia, acute myeloid leukemia, non-small cell lung cancer, and/or colon cancer. In an embodiment, said cancer is acute myeloid leukemia or breast cancer “Her2+”. 
     Other embodiments of this invention include a composition comprising a at least one of  Terminalia chebula  fruits,  Terminalia bellerica  fruits,  Phyllanthus emblica  fruits,  Withania somnifera  roots and leaves, Shilajit,  Azadirachta indica  leaves and twigs, or a standardized alcohol-water extract of the leaves, roots, or roots plus leaves of  Withania somnifera . In an embodiment, said  Terminalia chebula  fruit extract is AyuFlex®, said  Terminalia bellerica  fruit extract is Ayuric®, said  Phyllanthus emblica  fruit extract is Capros®, said  Withania somnifera  roots and/or leaves extract is Sensoril®, said Shilajit extract is PrimaVie®, and said  Azadirachta indica  leaves and twigs extract is PhytoBGS®. In an embodiment, said composition comprises a  Terminalia chebula  fruit extract and a  Terminalia bellerica  fruit extract, and optionally where said  T. chebula  extract is AyuFlex® and said  T. bellerica  extract is Ayuric®. In an embodiment, said composition comprises a  Terminalia chebula  fruit extract and a  Phyllanthus emblica  fruit extract, and optionally where said  T. chebula  extract is AyuFlex® and said  P. emblica  extract is Capros®. In an embodiment, said composition comprises a  Terminalia chebula  fruit extract and a  Withania somnifera  roots and leaves extract, or a standardized alcohol-water extract of the leaves, roots, or roots plus leaves of  Withania somnifera , and optionally where said  T. chebula  extract is AyuFlex® and said  W. somnifera  extract is Sensoril®. In an embodiment, said composition comprises a  Terminalia chebula  fruit extract and a Shilajit extract, and optionally where said  T. chebula  extract is AyuFlex® and said Shilajit extract is PrimaVie®. In an embodiment, said composition comprises a  Terminalia chebula  fruit extract and an  Azadirachta indica  leaves and twigs extract, and optionally where said  T. chebula  extract is AyuFlex® and said  A. indica  extract is PhytoBGS®. In an embodiment, said composition comprises a  Terminalia bellerica  fruit extract and a  Phyllanthus emblica  fruit extract, and optionally where said  T. bellerica  extract is Ayuric® and said  P. emblica  extract is Capros®. In an embodiment, said composition comprises a  Terminalia bellerica  fruit extract and a  Withania somnifera  roots and/or leaves extract, and optionally where said  T. bellerica  extract is Ayuric® and said  W. somnifera  extract is Sensoril®. In an embodiment, said composition comprises a  Terminalia bellerica  fruit extract and a Shilajit extract, and optionally where said  T. bellerica  extract is Ayuric® and said Shilajit extract is PrimaVie®. In an embodiment, said composition comprises a  Terminalia bellerica  fruit extract and an  Azadirachta indica  leaves and twigs extract, and optionally where said  T. bellerica  extract is Ayuric® and said  A. indica  extract is PhytoBGS®. In an embodiment, said composition comprises a  Phyllanthus emblica  fruit extract and a  Withania somnifera  roots and/or leaves extract, and optionally where said  P. emblica  extract is Capros® and said  W. somnifera  extract is Sensoril®. In an embodiment, said composition comprises a  Phyllanthus emblica  fruit extract and a Shilajit extract, and optionally where said  P. emblica  extract is Capros® and said Shilajit extract is PrimaVie®. In an embodiment, said composition comprises a  Phyllanthus emblica  fruit extract and an  Azadirachta indica  leaves and twigs extract, and optionally where said  P. emblica  extract is Capros® and said  A. indica  extract is PhytoBGS®. In an embodiment, said composition comprises a  Withania somnifera  roots and leaves extract and a Shilajit extract, and optionally where said  W. somnifera  extract is Sensoril® and said Shilajit extract is PrimaVie®. In an embodiment, said composition comprises a  Withania somnifera  roots and/or leaves extract and an  Azadirachta indica  leaves and twigs extract, and optionally where said  W. somnifera  extract is Sensoril® and said  A. indica  extract is PhytoBGS®. In an embodiment, said composition comprises a Shilajit extract and an  Azadirachta indica  leaves and twigs extract, and optionally where said Shilajit extract is PrimaVie® and said  A. indica  extract is PhytoBGS®. In an embodiment, said composition comprising Ayuflex® and Ayuric®. In an embodiment, said composition comprising Ayuflex® and Capros®. In an embodiment, said composition comprising Ayuric® and Capros®. In an embodiment, said composition comprising Ayuflex®, Ayuric®, and Capros®. In an embodiment, said composition comprising Sensoril® and PrimaVie®. In an embodiment, said composition comprising Sensoril® and Phyto-BGS®. In an embodiment, said composition comprising PrimaVie® and Phyto-BGS®. In an embodiment, said composition comprising Sensoril®, PrimaVie®, and Phyto-BGS®. In an embodiment, said composition comprises Crominex+3®. In an embodiment, said composition further comprises an anti-cancer drug; in an embodiment, said anti-cancer drug is pictilisib, doxorubicin, temozolomide, docetaxel, 5-fluorouracil (5-FU), ibrutinib, arsenic trioxide, and/or cytarabine, and/or any other anti-cancer drug, preferably identified in this application. In an embodiment, said composition comprises co-administration with an anti-cancer drug. In an embodiment, the present invention is directed to a method of treating cancer in a subject in need thereof, and/or enhancing the treatment of cancer in a subject in need thereof, comprising the steps of providing a composition comprising at least one extract, optionally a standardized aqueous extract that is Ayuflex®, Ayuric®, Capros®, Sensoril®, PrimaVie®, and/or PhytoBGS® or a hydroalcoholic extract that is Sensoril-AWE or a trivalent chromium complex that is Crominex+3®, and administering said composition in combination with an anti-cancer drug in an effective amount to treat and/or enhance treatment of cancer in the subject. In an embodiment, said anti-cancer drug is pictilisib, doxorubicin, temozolomide, docetaxel, 5-fluorouracil (5-FU), ibrutinib, arsenic trioxide, and/or cytarabine, and/or any other anti-cancer drug identified in this application. In an embodiment, a composition of the present invention comprises a combination of an anti-cancer drug, and in a separate composition, an extract. In an embodiment, the present invention is directed to a method of inhibiting the proliferation of cancer cells comprising the steps of providing a composition comprising at least one extract, preferably a standardized aqueous extract, preferably Ayuflex®, Ayuric®, Capros®, Sensoril®, PrimaVie®, and/or PhytoBGS®, and administering said composition in combination with an anti-proliferation drug in an effective amount to inhibit the proliferation of the cancer cells. 
     The use of the terms “a,” “an,” “the,” and similar referents in the context of describing the present invention (especially in the context of the claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. Use of the term “about” is intended to describe values either above or below the stated value in a range of approximately ±20%; in other embodiments, the values may range in value above or below the stated value in a range of approximately ±5%; in other embodiments, the values may range in value above or below the stated value in a range of approximately ±2%; in other embodiments, the values may range in value above or below the stated value in a range of approximately ±1%. The preceding ranges are intended to be made clear by context, and no further limitation is implied. All method steps described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise stated. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention. 
     While in the foregoing specification the present invention has been described in relation to certain embodiments thereof, and many details have been put forth for the purposes of illustration, it will be apparent to those skilled in the art that the invention is susceptible to additional embodiments and that certain of the details described herein can be varied considerably without departing from the basic principles of the invention. 
     The present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof, and, accordingly, reference should be made to the appended claims, rather than to the foregoing specification, as indicating the scope of the invention.