METHODS OF DETECTING AND TREATMENT OF CANCERS USING SCUTELLARIA BARBATA EXTRACT

BACKGROUND OF THE INVENTION

While advances in early detection and adjuvant therapy for breast cancer have had a favorable impact on patient survival in general, patients who develop advanced metastatic breast cancer are generally likely to face a less favorable prognosis. Commonly used hormonal and chemotherapeutic agents can lead to transient regression of tumors and can also palliate symptoms related to cancer. However, these treatments are often accompanied by toxicities and intolerable side effects and eventually become ineffective in controlling advanced stage breast cancer and its symptoms. Improvements in survival are modest, even with newer targeted biological agents. Moreover, in most metastatic cancers resistance to available conventional treatment ultimately develops or excessive side effects are seen with conventional therapies.

It is interesting to note that greater than 60% of all chemotherapeutic agents used in the treatment of breast cancer are derived from natural substances (Newman 2003). A fairly recent example is the development of taxanes from the Pacific yew tree, Taxus brevifolia. Throughout the world, it is estimated that approximately 80% of the world population still relies on botanical medicine as the primary source of therapy. In the West botanical medicine is considered a popular form of complementary and alternative medicine among patients diagnosed with cancer. However, few clinical trials have been conducted to firmly assess the safety and efficacy of botanical agents for the treatment of breast cancer, despite anecdotal case reports of cures and clinical efficacy in women who have relied solely on botanical medicine for treatment. It has previously been shown that the aqueous extract of Scutellaria Barbata can lead to growth inhibition of breast cancer cell lines in vitro (“Antiproliferative activity of Chinese medicinal herbs on breast cancer cells in vitro,” Anticancer Res., 22(6C):3843-52 (2002)). BZL101, a concentrated aqueous extract of Scutellaria Barbata, was evaluated for antiproliferative activity on five breast cancer cell lines (SK-BR-3, MCF7, MDA-MB-231, BT-474, and MCNeuA). These cell lines represent important prognostic phenotypes of breast cancer expressing a range of estrogen and HER2 receptors. BZL101, tested at a 1:10 dilution (15 μg/ml), demonstrated >50% growth inhibition on four of the five cell lines (Campbell, 2002). BZL101 showed >50% growth inhibition on a panel of lung, prostate and pancreatic cancer cell lines. BZL101 at the same dose did not cause >25% of growth inhibition on normal human mammary cells (HuMEC), demonstrating selectivity to cancer cells (Table 1). More so, BZL101 had a mild mitogenic effect on normal human lymphocytes. In cell cycle analysis, BZL101 caused an S phase burst and G1 arrest. BZL101 also attenuated mitochondrial membrane potential causing caspase-independent high molecular grade (HMG) apoptosis.

SUMMARY OF THE INVENTION

The inventor has recognized a need for improved methods of treating various types of cancer, especially ER⁻ (e.g. ERα⁻ and/or ERβ⁻) breast cancer. Various embodiments of the invention provided herein meet the foregoing need and provide related advantages as well.

Some embodiments described herein provide a method of treating cancer in a patient, comprising: obtaining a tumor sample from the patient; (a) contacting a portion of the tumor sample with a composition comprising an extract of Scutellaria barbata D. Don; (b) detecting a level of a marker of DNA oxidation in the sample from the patient; and (c) if the level of marker of DNA oxidation in the sample exceeds a predetermined threshold, administering to the patient an effective amount of an extract of Scutellaria Barbata D. Don. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is metastatic breast cancer. In some embodiments, the cancer is selected from the group consisting of sarcoma, carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, Kaposi's sarcoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma and retinoblastoma. In some embodiments, the marker of DNA oxidation is 8-oxoguanine or lactate. In some embodiments, the level of the marker of DNA oxidation is determined by mass spectrometry.

A method of deciding whether to continue anticancer chemotherapeutic treatment with an extract of Scutellaria barbata D. Don, comprising: (a) obtaining a sample from a cancer patient treated with an extract of Scutellaria barbata D. Don; (b) determining a level of a marker of DNA oxidation in the sample; and (c) if the level of marker of DNA oxidation in the sample exceeds a predetermined level, continuing treatment with the extract of Scutellaria barbata D. Don. In some embodiments, the cancer is a breast cancer. In some embodiments, the cancer is metastatic breast cancer. In some embodiments, the cancer is selected from the group consisting of sarcoma, carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, Kaposi's sarcoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma and retinoblastoma. In some embodiments, the marker of DNA oxidation is 8-oxoguanine or lactate. In some embodiments, the sample is a bodily fluid or a solid tissue. In some embodiments, the sample is a bodily fluid. In some embodiments, the bodily fluid is blood serum or urine. In some embodiments, the marker of DNA oxidation is 8-oxoguanine or lactate.

Some embodiments described herein provide a kit comprising a therapeutically effective amount of an extract of Scutellaria barbata D. Don and a means for determining a level of a marker of DNA oxidation in a sample from the patient. In some embodiments, the marker of DNA oxidation is 8-oxoguanine or lactate.

Some embodiments described herein provide a kit comprising a therapeutically effective amount of an extract of Scutellaria barbata D. Don and a means for detecting a level of a gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD, LTB4R, PBEF1, WDR3, NEK6, PLEKHF1, PLEK2, FKSG27, CORO1C, SNAI1, MOBKL1A, CYP27B1, PUS1, NFKBIB, UBE2E1, HA-1, PWP2H, TPST1, CDKN1A and ZNF529. In some embodiments, the gene participates in a xenobiotic response pathway, an oxidative response pathway, a NFκB pathway or an apoptosis/cell death pathway.

INCORPORATION BY REFERENCE

All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:

FIG. 1 shows dose-response curves showing the response of several solid cancer tumor cells to aqueous extract of the herb of this invention.

FIG. 2 shows dose-response curves showing the response of several breast solid cancer tumor cells to aqueous extract of the herb of the invention.

FIG. 3 shows dose-response curves comparing the response of breast solid cancer tumor cells and normal breast epithelium to aqueous extract of the herb of this invention.

FIG. 4 shows gel electrophoresis plate, which demonstrates that nuclear DNA disintegration occurs during apoptosis of solid tumor cancer cells in contact with aqueous extracts of the herb of this invention.

FIG. 5 shows the effect of the herb extract of the invention administered intraperitoneally (IP) on the tumors of mice in a xenograft model.

FIG. 6 shoes the effect of the herb extract administered by oral gavages and in interaction with cyclophosphamide administered in low dose in the drinking water on the tumors of mice in a xenograft model.

FIG. 7 shows that the herb extract induces apoptosis without activating caspases.

FIG. 8 shows that the herb extract in cell cycle analysis arrests the cells at the G1 phase.

FIG. 9 shows that illustrates that BZL101 leads to oxidative DNA damage. Formation of 8-oxoguanine, the most ubiquitous marker of DNA oxidation, was quantified through flow cytometric analysis of fixed permeabilized cells incubated with avidin fluorescein, that was shown to bind relatively specifically to 8-oxoguanine. There is a clear increase in binding of avidin to BZL101 treated SKBr3 cells versus untreated cells.

FIG. 10 shows that the conversion of non-fluorescent CM-H₂DCFDA into fluorescent compound is indeed due to ROS. Incubation of cells with ROS scavenger N-acetyl-cysteine (NAC) prior to addition of BZL101 prevented most of the increase in ROS generation.

DETAILED DESCRIPTION OF THE INVENTION

Some embodiments described herein provide a method of treating cancer in a patient, comprising: (a) obtaining a tumor sample from the patient; (b) contacting a portion of the tumor sample with a composition comprising an extract of Scutellaria barbata D. Don; (c) detecting a level of a gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don; and (d) if the level of the gene exceeds a predetermined threshold, administering to the patient an effective amount of an extract of Scutellaria Barbata D. Don. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is metastatic breast cancer. In some embodiments, the cancer is selected from the group consisting of sarcoma, carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, Kaposi's sarcoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma and retinoblastoma. In some embodiments, the gene is functionally involved in a xenobiotic response pathway, an oxidative response pathway, a NFκB pathway or an apoptosis/cell death pathway. In some embodiments, the gene is up-regulated by at least about 1.7 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD, LTB4R, PBEF1, WDR3, NEK6, PLEKHF1, PLEK2, FKSG27, CORO1C, SNAI1, MOBKL1A, CYP27B1, PUS1, NFKBIB, UBE2E1, HA-1, PWP2H, TPST1, CDKN1A and ZNF529. In some embodiments, the gene is up-regulated by at least about 1.8 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD, LTB4R, PBEF1, WDR3, NEK6, PLEKHF1, PLEK2, FKSG27, CORO1C, SNAI1, MOBKL1A, CYP27B1, PUS1, NFKBIB, UBE2E1 and HA-1. In some embodiments, the gene is up-regulated by at least about 1.9 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD and LTB4R. In some embodiments, the gene is up-regulated by at least about 2.0 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2 and TUBB2. In some embodiments, the gene is up-regulated by at least about 2.1 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL and SLC2A6. In some embodiments, the gene is up-regulated by about 2.2 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1 and CXCL16. In some embodiments, the gene is up-regulated by at least about 2.7 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6 and BBC3. In some embodiments, the gene is a gene involved in a cellular xenobiotic response. In some embodiments, the gene is CYP1A1, CYP1B1, HSPA6, CYP27B1. In some embodiments, the gene is a gene involved in the oxidative response pathway. In some embodiments, the gene is selected from the group consisting of HMOX1, TNFAIP3, OKL38, GCLM, CBS, ATF3, and TXNRD1. In some embodiments, the gene is involved in the NFκB pathway. In some embodiments, the gene is selected from the group consisting of HMOX1, CXCL1, CYP1B1, TNFAIP3, IGFL1, TNF, CLC, BIRC3, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, BBC3, ATF3, GADD45A, CCL11, RASD1, NFKBIE, PANX1, IRF1, TRAF3, EDN1, PBEF1, NEK6, NFKBIB, TPST1 and CDKN1A. In some embodiments, the gene is involved in the apoptosis/cell death pathway. In some embodiments, the gene is selected from the group consisting of TNFAIP3, TNF, BIRC3, CCL2, SQSTM1, CLC, TNFRSF21, BBC3, GADD45A, SAT, CCL11, NFKBIE, TRAF3, MMD and CDKN1A. In some embodiments, the method further comprises detecting a level of a second gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don. In some embodiments, the method further comprises detecting a level of a third gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don.

Some embodiments provide method of deciding whether to continue anticancer chemotherapeutic treatment with an extract of Scutellaria barbata D. Don, comprising: (a) obtaining a sample from a cancer patient treated with an extract of Scutellaria barbata D. Don; (b) determining a level gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don in the sample; and (c) if the level of gene that is up-regulated exceeds a predetermined level, continuing treatment with the extract of Scutellaria barbata D. Don. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is metastatic breast cancer. In some embodiments, the cancer is selected from the group consisting of sarcoma, carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, Kaposi's sarcoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma and retinoblastoma. In some embodiments, the gene is functionally involved in a xenobiotic response pathway, an oxidative response pathway, a NFκB pathway or an apoptosis/cell death pathway. In some embodiments, the gene is up-regulated by at least about 1.7 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD, LTB4R, PBEF1, WDR3, NEK6, PLEKHF1, PLEK2, FKSG27, CORO1C, SNAI1, MOBKL1A, CYP27B1, PUS1, NFKBIB, UBE2E1, HA-1, PWP2H, TPST1, CDKN1A and ZNF529. In some embodiments, the gene is up-regulated by at least about 1.8 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD, LTB4R, PBEF1, WDR3, NEK6, PLEKHF1, PLEK2, FKSG27, CORO1C, SNAI1, MOBKL1A, CYP27B1, PUS1, NFKBIB, UBE2E1 and HA-1. In some embodiments, the gene is up-regulated by at least about 1.9 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD and LTB4R. In some embodiments, the gene is up-regulated by at least about 2.0 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2 and TUBB2. In some embodiments, the gene is up-regulated by at least about 2.1 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL and SLC2A6. In some embodiments, the gene is up-regulated by about 2.2 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1 and CXCL16. In some embodiments, the gene is up-regulated by at least about 2.7 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6 and BBC3. In some embodiments, the gene is a gene involved in a cellular xenobiotic response. In some embodiments, the gene is CYP1A1, CYP1B1, HSPA6, CYP27B1. In some embodiments, the gene is a gene involved in the oxidative response pathway. In some embodiments, the gene is selected from the group consisting of HMOX1, TNFAIP3, OKL38, GCLM, CBS, ATF3, and TXNRD1. In some embodiments, the gene is involved in the NFκB pathway. In some embodiments, the gene is selected from the group consisting of HMOX1, CXCL1, CYP1B1, TNFAIP3, IGFL1, TNF, CLC, BIRC3, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, BBC3, ATF3, GADD45A, CCL11, RASD1, NFKBIE, PANX1, IRF1, TRAF3, EDN1, PBEF1, NEK6, NFKBIB, TPST1 and CDKN1A. In some embodiments, the gene is involved in the apoptosis/cell death pathway. In some embodiments, the gene is selected from the group consisting of TNFAIP3, TNF, BIRC3, CCL2, SQSTM1, CLC, TNFRSF21, BBC3, GADD45A, SAT, CCL11, NFKBIE, TRAF3, MMD and CDKN1A. In some embodiments, the method further comprises detecting a level of a second gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don. In some embodiments, the method further comprises detecting a level of a third gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don.

Some embodiments described herein provide a method of treating cancer in a patient, comprising: (a) treating the patient with a first dosage of extract of Scutellaria barbata D. Don; (b) obtaining a sample from the patient; (c) detecting in the sample a level of a gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don; (d) if the level of the gene that is up-regulated exceeds a predetermined level, continuing treatment with the first dosage of extract of Scutellaria barbata D. Don; and (e) if the level of the gene does not exceed the predetermined level, discontinuing treatment with Scutellaria barbata D. Don. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is metastatic breast cancer. In some embodiments, the cancer is selected from the group consisting of sarcoma, carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, Kaposi's sarcoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma and retinoblastoma. In some embodiments, the gene is functionally involved in a xenobiotic response pathway, an oxidative response pathway, a NFκB pathway or an apoptosis/cell death pathway. In some embodiments, the gene is up-regulated by at least about 1.7 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD, LTB4R, PBEF1, WDR3, NEK6, PLEKHF1, PLEK2, FKSG27, CORO1C, SNAI1, MOBKL1A, CYP27B1, PUS1, NFKBIB, UBE2E1, HA-1, PWP2H, TPST1, CDKN1A and ZNF529. In some embodiments, the gene is up-regulated by at least about 1.8 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD, LTB4R, PBEF1, WDR3, NEK6, PLEKHF1, PLEK2, FKSG27, CORO1C, SNAI1, MOBKL1A, CYP27B1, PUS1, NFKBIB, UBE2E1 and HA-1. In some embodiments, the gene is up-regulated by at least about 1.9 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD and LTB4R. In some embodiments, the gene is up-regulated by at least about 2.0 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2 and TUBB2. In some embodiments, the gene is up-regulated by at least about 2.1 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL and SLC2A6. In some embodiments, the gene is up-regulated by about 2.2 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1 and CXCL16. In some embodiments, the gene is up-regulated by at least about 2.7 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6 and BBC3. In some embodiments, the gene is a gene involved in a cellular xenobiotic response. In some embodiments, the gene is CYP1A1, CYP1B1, HSPA6, CYP27B1. In some embodiments, the gene is a gene involved in the oxidative response pathway. In some embodiments, the gene is selected from the group consisting of HMOX1, TNFAIP3, OKL38, GCLM, CBS, ATF3, and TXNRD1. In some embodiments, the gene is involved in the NFκB pathway. In some embodiments, the gene is selected from the group consisting of HMOX1, CXCL1, CYP1B1, TNFAIP3, IGFL1, TNF, CLC, BIRC3, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, BBC3, ATF3, GADD45A, CCL11, RASD1, NFKBIE, PANX1, IRF1, TRAF3, EDN1, PBEF1, NEK6, NFKBIB, TPST1 and CDKN1A. In some embodiments, the gene is involved in the apoptosis/cell death pathway. In some embodiments, the gene is selected from the group consisting of TNFAIP3, TNF, BIRC3, CCL2, SQSTM1, CLC, TNFRSF21, BBC3, GADD45A, SAT, CCL11, NFKBIE, TRAF3, MMD and CDKN1A. In some embodiments, the method further comprises detecting a level of a second gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don. In some embodiments, the method further comprises detecting a level of a third gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don.

Some embodiments described herein provide a method of treating cancer in a patient, comprising: (a) treating the patient with a first dosage of extract of Scutellaria barbata D. Don; (b) obtaining a sample from the patient; (c) detecting in the sample a level of a gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don; (d) if the level of the a level of the gene that is up-regulated exceeds a predetermined level, continuing treatment with the first dosage of extract of Scutellaria barbata D. Don; and (e) if the level of the gene does not exceed the predetermined level, discontinuing treatment with Scutellaria barbata D. Don. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is metastatic breast cancer. In some embodiments, the cancer is selected from the group consisting of sarcoma, carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, Kaposi's sarcoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma and retinoblastoma. In some embodiments, the gene is functionally involved in a xenobiotic response pathway, an oxidative response pathway, a NFκB pathway or an apoptosis/cell death pathway. In some embodiments, the gene is up-regulated by at least about 1.7 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD, LTB4R, PBEF1, WDR3, NEK6, PLEKHF1, PLEK2, FKSG27, CORO1C, SNAI1, MOBKL1A, CYP27B1, PUS1, NFKBIB, UBE2E1, HA-1, PWP2H, TPST1, CDKN1A and ZNF529. In some embodiments, the gene is unregulated by at least about 1.8 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD, LTB4R, PBEF1, WDR3, NEK6, PLEKHF1, PLEK2, FKSG27, CORO1C, SNAI1, MOBKL1A, CYP27B1, PUS1, NFKBIB, UBE2E1 and HA-1. In some embodiments, the gene is up-regulated by at least about 1.9 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2, TUBB2, CPEB2, FADS3, NCOA7, TRAF3, TRIM21, EGR1, MOBKL2C, PEO1, DDX31, TXNRD1, EDN1, TIGA1, TM4SF14, AXIN1, MMD, CEBPD and LTB4R. In some embodiments, the gene is up-regulated by at least about 2.0 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4AT, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL, SLC2A6, TUBB6, RCL1, ABCC2, LRFN1, MGC35521, IRF1, SAT, OPTN, SESN2 and TUBB2. In some embodiments, the gene is up-regulated by at least about 2.1 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1, CXCL16, KRT5, MYEOV, SNAPC4, ERBP, UBE2E2, ZFP36, PLK2, SQRDL and SLC2A6. In some embodiments, the gene is up-regulated by about 2.2 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6, BBC3, ELL2, ATF3, C20orf139, GADD45A, SAT, SLCO4A1, NKX3-1, CCL11, MGAM, RASD1, ZSWIM4, NFKBIE, P53AIP1, NICAL, CCRN4L, RAPGEFL1, CA8, ARRDC2, NELF, PHLDA2, CLDN1, IER5, PIK3CD, PANX1 and CXCL16. In some embodiments, the gene is up-regulated by at least about 2.7 fold over cellular expression in the absence of Scutellaria barbata D. Don. In some embodiments, the gene is selected from the group consisting of CYP1A1, HMOX1, LOC255324, CXCL1, CYP1B1, LOC440449, TNFAIP3, IGFL1, NPAS2, TNF, CLC, BIRC3, OKL38, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, CDH5, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, RNF24, AMSH-LP, ADM, HSPA6 and BBC3. In some embodiments, the gene is a gene involved in a cellular xenobiotic response. In some embodiments, the gene is CYP1A1, CYP1B1, HSPA6, CYP27B1. In some embodiments, the gene is a gene involved in the oxidative response pathway. In some embodiments, the gene is selected from the group consisting of HMOX1, TNFAIP3, OKL38, GCLM, CBS, ATF3, and TXNRD1. In some embodiments, the gene is involved in the NFκB pathway. In some embodiments, the gene is selected from the group consisting of HMOX1, CXCL1, CYP1B1, TNFAIP3, IGFL1, TNF, CLC, BIRC3, ICAM1, IL8, RELB, CCL2, SQSTM1, CLC, TIPARP, CCL2, PLAUR, TNFRSF21, GCLM, CBS, BBC3, ATF3, GADD45A, CCL11, RASD1, NFKBIE, PANX1, IRF1, TRAF3, EDN1, PBEF1, NEK6, NFKBIB, TPST1 and CDKN1A. In some embodiments, the gene is involved in the apoptosis/cell death pathway. In some embodiments, the gene is selected from the group consisting of TNFAIP3, TNF, BIRC3, CCL2, SQSTM1, CLC, TNFRSF21, BBC3, GADD45A, SAT, CCL11, NFKBIE, TRAF3, MMD and CDKN1A. In some embodiments, the method further comprises detecting a level of a second gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don. In some embodiments, the method further comprises detecting a level of a third gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don.

Pharmaceutical Compositions and Modes of Administrations

An extract of this invention can be administered to a patient either as a “tea,” without combination with any other substances or further manipulation, or it can be administered as a pharmaceutical composition where the extract is mixed with suitable carriers or recipient(s). In treating a patient exhibiting a disorder of interest, a therapeutically effective amount of the extract is administered. A therapeutically effective amount refers to that amount of the extract that results in amelioration of symptoms or a prolongation of survival in a patient, and may include destruction of a malignant tumor of a microbial infection.

When administered without combination with any other substances, the composition comprising extract of Scutellaria Barbata (especially Scutellaria Barbata D. Don) may be encased in a suitable capsule, such as a gelatin capsule. When administered in admixture with other excipients, adjuvants, binders, diluents, disintegrants, etc., the dry extract of Scutellaria Barbata may be compressed into a capsule or caplet in a conventional manner that is well-known in the art.

Toxicity and therapeutic efficacy of the extracts, i.e., determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population) can be determined by standard pharmaceutical procedures in cell cultures or experimental animals. The dose ratio between toxic and therapeutic effects is therapeutic index and it can be expressed as the ratio LD50/ED50. Extracts that exhibit large therapeutic indices are preferred. The data obtained from these cell culture assays and animal studies can be used in formulating a range of dosages for use in humans, in particular for internal use, that include ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. In general, since the extracts used in the methods of this invention have been used in TCM, they are known to be relatively non-toxic to humans and therefore it is expected that they will exhibit large therapeutic indices.

For any extract used in the method of invention, therapeutically effective dose can be estimated initially from cell culture assays. For example, a dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by HPLC.

The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition and based on knowledge of TCM. (See e.g. Fingl et al., in The Pharmacological Basis of Therapeutics, 1975, Ch. 1, p. 1). It should be noted that the attending physician would know how and when to terminate, interrupt, or adjust administration due to toxicity, or organ dysfunction. Conversely, the attending physician would also know to adjust treatment to higher levels if the clinical response is not adequate. The severity of the condition may, for example, be evaluated, in part, by standard prognostic evaluation methods. Further, the dose and perhaps dose frequency will also vary according to the age, body weight, and response of the individual patient. A program comparable to that discussed above may be used in veterinary medicine.

If desired, standard western medicine techniques for formulation and administration may be used, such as those found in Remington's Pharmaceutical Sciences, 18^thed., Mack Publishing Co., Easton, Pa. (1990). Suitable routes may include: oral, rectal, transdermal, vaginal, transmucosal, or intestinal administration; parenteral delivery, including intramuscular, subcutaneous, intramedullary injections; as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, or intraocular injections, to name a just a few. In particular embodiments, the extract of the invention is administered orally.

For injection, an extract of this invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer, For such transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

Use of pharmaceutically acceptable carriers to formulate an extract herein use in the methods disclosed for the practice of this invention in dosages suitable for systemic administration is within the scope of the invention. With proper choice of carrier and suitable manufacturing practice, an extract of the present invention, in particular those formulated as solutions, may be administered parenterally, such as by intravenous injection. Likewise, an extract can be formulated, using pharmaceutically acceptable carriers well known in the art, into dosages suitable for oral administration. Such carriers enable extracts to be formulated as tablets, pills, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated.

Pharmaceutical compositions suitable for use in the present invention are compositions wherein an extract is contained in an effective amount to achieve its intended purpose. Determination of the effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein. A pharmaceutical composition may contain suitable pharmaceutically acceptable carriers including excipients and auxiliaries that facilitate processing of the extracts into preparations that can be used pharmaceutically. The preparations formulated for oral administration may be in the form of tablets, dragees, capsules, or solutions. The pharmaceutical compositions of the present invention may be manufactured in a manner that is itself known, e.g., by means of convention mixing, dissolving, granulating, dragees, capsules, or solutions. The pharmaceutical compositions of the present invention may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levitating, emulsifying, encapsulating, entrapping or lyophilizing processes.

Pharmaceutically formulations for parenteral administration include aqueous solutions of an extract in water-soluble form. Additionally, suspensions of an extract may be prepared as appropriate oily injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of an extract to allow for the preparation of highly concentrated solutions.

Pharmaceutical preparations for oral use can be obtained by combining an extract with solid excipient, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum Arabic, talc, polyvinyl pyrrolidone, carpool gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of extracts and/or doses.

Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules contain the extract in admixture with fillers such as lactose, binders such as starches, and/or lubricants such as talc or magnesium separate and, optionally, stabilizers. In soft capsules, the extract may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols.

The dosage of extract of Scutellaria barbata D. Don will vary depending upon the tumor type, the stage of disease, the species of patient and the individual patient. In some embodiments, the amount of extract of Scutellaria barbata D. Don (BZL) administered to a human patient will be the dry solid residue extracted from about 0.1 g to about 20,000 g of dried solid plant parts of BZL. In some embodiments, the effective dose is the dry solid residue extracted from about 1 to about 1000 g of BZL. In some embodiments, the effective dose will be the dry solid residue extracted from about 10 to about 800 g of BZL.

Treatment of Cancers

Extracts of Scutellaria barbata D. Don may be used to treat solid tumors. Such tumors may include so-called estrogen receptor negative (ER⁻) breast cancer, estrogen receptor positive (ER⁺) cancer, and other solid tumor cancers. As used herein, the terms “estrogen receptor negative breast cancer” and “estrogen receptor positive breast cancers,” have meanings commonly ascribed to them in the art. The person skilled in the art will recognize that the terms “positive” and “negative” are relative terms describing levels of expression in a cell. In general, saying that a cell is “negative” for expression of a particular cell product means that the level of expression detected, if any, falls below a predetermined threshold. That threshold may be a detection limit, a background noise level or some arbitrary cutoff known and understood by one of skill in the art. As extracts of Scutellaria barbata D. Don do not necessarily require presence of ERα or ERβ in order to induce apoptosis in solid cancer cells, it is considered that doses of Scutellaria barbata D. Don may be used to treat, inter alia, either ER⁺ or ER⁻ breast cancers as well as other solid tumors. The dose of Scutellaria barbata D. Don extract may vary, however it is considered that a dose comprising the dry soluble portion of a hot water or ethanolic extract of about 1 to about 20,000 g, especially about 50 to about 10,000 g of dry aerial portions of Scutellaria barbata D. Don, is a therapeutically effective dose. When used in combination with another chemotherapeutic agents, the dose may be lowered to take advantage of synergetic effects. C that extracts of Scutellaria barbata D. Don may be used to treat include sarcoma, carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, Kaposi's sarcoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma and retinoblastoma. In some particular embodiments, the

Kits

Also provided herein are kits for treatment of cancer. In some embodiments, the kits comprise an extract of Scutellaria barbata D. Don. In some embodiments, a the extract of Scutellaria barbata D. Don is in an oral dosage form. In some embodiments, the kit will contain sufficient extract of Scutellaria barbata D. Don for administration over 1, 2, 3, 4 or more weeks. In some embodiments, the dosage of extract of Scutellaria barbata D. Don will be divided into daily or twice daily doses. The daily dose of extract of Scutellaria barbata D. Don may vary depending on the second chemotherapeutic agent, the disease to be treated, the condition of the patient, etc. In general, the daily dose of extract of Scutellaria barbata D. Don will be the dried soluble extract of about 1 to 20,000 g, 10 to 10,000 g or 50 to 5000 g of dried aerial portion of Scutellaria barbata D. Don. The daily dose may be divided into 2, 3, 4 or more doses per day. When administered as a tea, the doses may be combined with a flavor or flavor-masking agent in order to enhance palatability.

Some embodiments described herein provide a kit for treatment of cancer, comprising a therapeutically effective amount of a first chemotherapeutic agent comprising an extract of Scutellaria Barbata D. Don and a means for testing a level of expression of a gene that is up-regulated in a cell in which an extract of Scutellaria barbata D. Don is cytotoxic and which has been contacted with Scutellaria barbata D. Don. The means for testing may include reagents and/or instructions for work-up methods for preparing a sample for evaluation on a gene chip or by mass spectrometry or both. The means may include antibodies (including labeled antibodies) for ELISA or similar methods. The means may include PCR probes and/or mass spectrometry standards for mass spectrometry measurements.

EXAMPLES

The herb from which the extracts of this invention were obtained were purchased from Shen Nong Herbs, Berkeley, Calif. Their identity was confirmed by reference to traditional pharmaceutical literature.

Preparative Example 1
Preparation of BZL101 for in vitro and Mouse Experiments

Herbal extract was prepared as “boiled teas”, which is how most are prepared for use in traditional treatment regimes. Aqueous extracts were prepared by adding 7.5 g of dry ground herb to 125 ml distilled water, bringing the mixture to a boil and then simmering for 45 minutes, The mixture was cooled, during which period most of the solids sank to the bottom of the vessel. The aqueous layer was carefully decanted off of the residual solids, centrifuged for 5 minutes at 1500 rpm, sterile filtered through a 0.45 μm filter and stored at 4° C. until used. Generally, the extracts were tested within 1-2 weeks of preparation although most of the active extracts were found to retain activity after storage at 4° C. for several additional weeks. An aliquot of each extract was dried under vacuum and the dry weight of the water soluble substances extracted from each herb determined.

Preparative Example 2
Preparation of BZL101 for Human in vivo Experiments

BZL101 is an aqueous extract of the aerial part of Scutellaria Barbata D. Don of the Lamiaceae family. Herba Scutellaria Barbata D. Don (Chinese pin yin transliteration—Ban Zhi Lian (BZL)) is grown mainly in areas southeastern of the Yellow River (Huang Po) in the provinces of Sichuan, Jiangsu, Jiangxi, Fujian, Guangdong, Guangxi and Shaanxi. The plant is harvested in late summer and early autumn after it blooms. The aerial part (leaves and stems) is cut from the root and is used as starting material (BZL). The aerial part of the herb is dried in the sun, packed as a whole plant. The herb is identified and verified through botanical, morphological and chemical characteristics to ensure purity.

A single dose of BZL101 is made through the following procedure and is termed BZL101 (Bionovo, Inc., Emeryville, Calif.).

- 180 grams of the raw herb is ground to fine powder (25 mesh)
- The powder is mixed with 1800 ml of distilled water to form a slurry
- The slurry is than simmered at 70-72° C. for 60 minutes
- The extract is decanted and filtered through 22 μm filter
- The supernatant weight after extraction is 168 gm
- The volume of the solution is 1750 ml
- The extract is concentrated with a vacuum evaporator to reduce the volume of water to 350 ml which constitutes a 5:1 concentration of the original solution
- The dry weight of soluble material in the extract is 12 gm
- It is packaged in a sterile, vacuum sealed container
- Testing for bacteria, yeast and heavy metals are preformed by an accredited laboratory

Comparative Example 1
In vitro Inhibition of Cancer Cell Activity Cell Lines and Culture

The extract obtained in Preparative Example 1, above, was tested against four human breast cancer cell lines, SKBR3, MFC-7, MDA-MB231 and BT474, and one murine breast cancer cell line, MCNeuA. All lines were maintained in 90% DME supplement with 2.0 mom L-glutamine, 100 IU/ml penicillin, 100 μg/ml streptomycin and 10% heat-inactivated fetal bovine serum. Cells at 70-80% confluence were used for plating for growth inhibition assays.

Cells were plated in 96-well flat bottom plates at 5,000 to 10,000 cells/well. The difference in number of cells plated adjusts for differences in the growth rates of these cell lines. Cells were allowed to adhere to the well walls overnight; then the extracts were added to triplicate wells at a 1:10 final dilution in culture medium for initial screening. For generating dose-response curves, serial 3-fold dilutions, starting at 1:10 dilution over 6 rows of wells were used. Water was added to the control wells at 1:10 dilution in culture medium. The plates were incubated at 37° C., 5% CO₂, for 3 days and then assayed for growth inhibition using a crystal violet assay (Bernhardt, G., et al., Standardized Kinetic Microassay to Quantify Differential Chemosensitivity on the Basis of Proliferative Activity, 1992, J. Cancer Res. Clin. Oncol., 118:35-43). Cells remaining adherent to the well walls were rinsed with PBS, the fixed cells were stained with 0.02% aqueous crystal violet (50 μl/well) for 30 minutes after which the wells were washed thoroughly with distilled water. The crystal violet stain bound by the cells was solubilized in 79% ethanol (100 μl/well) and the plates analyzed on a microplate reader (Molecular Devices) ay 595 nm. The percent inhibition was calculated as the average optical density of the control wells minus average optical density extract well divided by the average optical density of the control wells. Dose-response curves on SKBR3, MCF7 and MCNeuA cells for several of the extracts are shown in FIGS. 1-3. As can be seen, the concentration at which the extracts inhibited the activity of the cells by 50% (the IC50) ranged from over 1 mg/ml down to about 10 μg/ml.

Induction of Apoptosis

To assay for DNA fragmentation as a marker of apoptosis, a procedure for the isolation of genomic DNA that allows for the analysis of both high and low molecular weight DNA fragmentation during apoptosis was used. MCNeuA cells were plated at 5×10⁵cells/well in 6-plates and allowed to adhere overnight. Aqueous herbal extracts were added to each well at a 1:10 and a 1:50 dilution. Sterile water, diluted 1:10 in culture medium, was added to the control wells. After 24 hours, the cells were visually examined under a microscope and morphological changes noted. Attached and floating cells were harvested, washed with cold PBS and embedded in lysis buffer (50 mM NaCl, 20 mM Tris HCl, pH 8.0, 20 mM EDTA, 0.5% sodium sarkosyl, 50 μg/ml Rnase A and 100 μg/ml proteinase K) for 1 hour at 37° C. The cells were then washed with PBS and distilled water and placed in the wells of a conventional 1% agarose gel and electrophoresed overnight at approximately 1 V/cm. The gels were then stained with ethidium bromide and photographed under UV transillumination to give intense images. The images obtained are shown in FIG. 4.

BZL101 was evaluated for antiproliferative activity on five breast cancer cell lines (SK-BR-3, MCF7, MDA-MB-231, BT-474, and MCNeuA). These cell lines represent important prognostic phenotypes of breast cancer expressing a range of estrogen and HER2 receptors. BZL101, tested at a 1:10 dilution (15 μg/ml), demonstrated >50% growth inhibition on four of the five cell lines (Campbell, 2002). BZL101 showed >50% growth inhibition on a panel of lung, prostate and pancreatic cancer cell lines. BZL101 at the same dose did not cause >25% of growth inhibition on normal human mammary cells (HuMEC), demonstrating selectivity to cancer cells (Table 3). Moreso, BZL101 had a mild mitogenic effect on normal human lymphocytes. In cell cycle analysis, BZL101 caused an S phase burst and G1 arrest. (See FIG. 8). BZL101 also attenuated mitochondrial membrane potential causing caspase-independent high molecular grade (HMG) apoptosis. (See FIG. 7).

The results of this in vitro experiment are summarized in Table 3, below.

TABLE 3

In vitro growth inhibitory effect of BZL101 aqueous extract of Scutellaria Barbata 1:10

dilution− < 50% inhibition, + 51-75% inhibition, ++ > 75% inhibition.

BZL is active on all cancer cell lines but is not active on HuMECs.

Breast

Pancreas

MDA-

Lung

Panc
Prostate

MB-

A549
LLC
Panc-1
02
PC-3
LNCaP
MCF7
BT474
SKBR3
231
MCNeuA
HuMEC

+
+
+
++
+
+
++
+
++
+
++
−

Example 1
Methods of Detecting Urinary Analysis of 8-oxoguanine, 8-oxoguanosine, fapy-guanine and 8-oxo-2′-deoxyguanosine as a Biomarker of Efficacy in the Treatment of BZL101 in Patients with Adenocarcinoma

Rationale

Reactive oxygen species (ROS) have been strongly associated with cellular aging, cancer, and other degenerative diseases by virtue of their potential to damage several cellular constituents, such as nucleic acids, proteins, and lipids. It is now well established that free radical mediated oxidation of DNA leads to a broad spectrum of chemical modifications which translate into single or double DNA strand breaks, and base as well as sugar modifications. Among the DNA base modifications induced by oxidative damage, 8-oxoguanine (8-oxoGua) is of particular relevance and has been proposed as a biomarker of DNA oxidation. The DNA guanine base oxidation product 8-oxo-2′-deoxyguanosine (8-oxodG) is potentially mutagenic and commonly quantified as a steady-state estimate of oxidative stress in tissues and urine using chromatographic techniques.

The urinary excretion of products of damaged nucleotides in cellular pools or in DNA may be important biomarkers of exposure to relevant carcinogens and may predict cancer risk or response to treatment. It is important to recognize that in steady state the excretion reflects the rate of damage. Among the many oxidative DNA damage products 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) is probably the most studied because of the relative ease of measurement and the mutagenic properties resulting in G→T transversion mutations upon replication of DNA.

Oxidized guanine in DNA is mainly repaired by oxoguanine glycosylase (OGG1) resulting in release of 8-oxoguanine. This enzyme shows a common genetic polymorphism with a variant Ser326Cys, which in complementation assays in vitro appears to increase susceptibility to mutagenic properties of ROS considerably, whereas 8-oxodG levels and incision activity in leukocytes and some target tissues generally show no difference between the genotypes. In addition, repair of 8-oxodG may to some extent occur by nucleotide excision repair and transcription coupled mechanisms. A specialized enzyme (MTH1 or NUDT1) sanitizes the nucleotide pool by cleaving phosphates of 8-oxodGTP which if incorporated during DNA synthesis is highly mutagenic and mice deficient in that enzyme develop tumors. 8-OxodG from this process as well as from putative nucleotide excision repair and possibly mitochondrial turn-over is excreted unchanged into the urine and may serve as a biomarker of oxidative stress and oxidative damage to nucleotides and possibly DNA. The urinary excretion of 8-oxodG has consistently been found to be increased among smokers and with a number of occupational exposures, including air pollution among bus drivers. Moreover, some case—control studies have suggested that the urinary excretion of 8-oxodG or 8-oxoguanine is increased among cancer patients, although this could very well be a consequence of the disease with ongoing oxidative stress, inflammation and tissue turn-over.

Furthermore, a great emphasis has being placed on the role of ROS-induced DNA damage in carcinogenesis and aging as a consequence of genomic degradation.

In recent years, there has been an increased interest in the use of mass spectrometry (MS) for the analysis of DNA oxidation products. MS typically provides structural information and selective detection in the picogram to femtogram range.

Therefore, given these characteristics, mass spectrometry is well positioned to play a significant role in the detection and characterization of DNA adducts. Besides the well-established role of DNA oxidative damage in many disease conditions, growing evidence points towards a significant involvement of RNA oxidative damage in the pathophysiology of several age-related degenerative disorders including cancer. Indeed, although little is still known about the consequences of RNA oxidation on the cellular homeostasis, it has been recently shown that oxidized RNA is associated with impaired protein synthesis as a consequence of translation errors. Moreover, RNA has been found to be significantly oxidized in age-related degenerative diseases.

BZL101 and 8-oxo-guanine

It is considered that differential induction of DNA damage by BZL101 in different cell types might be related to the extent of oxidative stress generated by BZL101 treatment. BZL101 induced a significant accumulation of ROS in SKBr3 cells as measured by staining with the ROS-sensitive probe CM-H₂DCFDA. Incubation of cells with ROS scavenger N-acetyl-cysteine (NAC) prior to addition of BZL101 has prevented most of the increase in ROS generation, confirming that the conversion of non-fluorescent CM-H₂DCFDA into fluorescent compound is indeed due to ROS. To confirm that BZL101 induces oxidative stress responses, the levels of transcriptional factor Nrf2 in BZL101 treated cells were examined. Nrf2 is a key regulator of phase II detoxifying and antioxidant enzymes that are upregulated in response to oxidative stress. Western blot analysis showed a significant and sustained increase in Nrf2 levels in BZL101 treated BT474 cells and SKBr3 cells. In MCF10A cells, there was also an increase in Nrf2 levels, though it was more transient in nature.

There is a different fold increase of ROS levels in different cells treated with BZL101 compared to control untreated cells. It is of particular interest that the increase in ROS correlates well with the degree of DNA damage induced in these cells. The lowest induction is seen in MCF10A cells which also have the lowest number of comets after treatment with BZL101, and the highest increase in ROS is observed in SKBr3 cells where the DNA damage is most extensive. In fibroblasts IMR90 and in BT474 cells the moderate increase in ROS paralleled the relatively lower extent of DNA damage compared to SKBr3.

To further implicate ROS in the induction of DNA damage, comet formation in cells pretreated with the antioxidants NAC and pyruvate prior to the addition of BZL101 were examined. Both compounds have significantly reduced the number of cells forming comets, in particular in normal cell lines. DNA damage repair in cancer cells in the presence of NAC or pyruvate was also greatly accelerated (not shown). At the same time, pretreatment of cells with the nitric oxide scavenger PTIO had no effect on the numbers of cells with comets (not shown), indicating that most of DNA lesions induced by BZL101 are oxidative in nature.

To verify that BZL101 leads to oxidative DNA damage, it was determined whether the DNA of BZL101-treated cells contains 8-oxoguanine, the most ubiquitous marker of DNA oxidation. Formation of 8-oxoguanine has been quantified through flow cytometric analysis of fixed permeabilized cells incubated with avidin fluorescein, that was shown to bind relatively specifically to 8-oxoguanine. There is a clear increase in binding of avidin to BZL101 treated SKBr3 cells versus untreated cells. (See FIG. 9). This increase was completely abolished if cells were pretreated with NAC prior to addition of BZL101, confirming the specificity of observed staining. (See FIG. 10).

Analysis of the 8-oxoguanidine and apurininc/apyrimidinic (AP) bases in DNA. The Calbiochem kit was used for staining of the fixed and permeabilized cells with avidin fluorescein. Quantification of apurinic/apyrimidininc nucleotides in genomic DNA was performed using the DNA damage quantification kit from BioVision Research Products.

Example 4
Micro Array Gene Expression Signature Profiling for Patient Selection for Treatment with BZL101

A micro array gene expression analysis was performed, using Phalanx human gene chip containing 45,000 gene probes of 36,000 unique human genes. SKBR3 and BT474 breast cancer cells were treated with BZL101 for 18 hours and compared with untreated control cells.

The mRNAs that are induced by BZL101 was grouped into functional groups. Only RNAs induced at higher that 2.5 fold were considered. Among these genes there is only one that is difficult to assign (AMSH-LP).

There are very few functions that are affected by BZL, and they are:

Apoptosis

Cytokines with proliferative effects

NF kappa B pathway

Oxidative stress

DNA damage response

Cell adhesion

There are obviously overlaps between these groups. Many of cytokines could go into NFkB pathway group. Virtually all oxidative stress responders could also go into NFkB pathway group because they are induced by transcription factor NRF2 with the involvement of NFkB. A number of genes in different groups are induced by TNF. DNA damage responders could go into oxidative stress group because they are induced by oxidative DNA damage.

The oxidative stress responders have links to the glycolytic pathway and DNA damage.

Apoptosis

- TNFAIP3=A20 (six fold)
- TNFRSF21=death receptor 6 (3 fold)
- TNF (4.7 fold)
- BBC3=PUMA (2.7 fold in SKbr3)

Cytokines with Proliferative Activities

Chemokine CXCL1(melanoma growth factor); seven-fold

Insulin growth factor like family IGFL1 (five fold)

Cardiotropin-like cytokine NNT-1 or CLC (4.6 fold) twice

Chemokine ligand 2 CCL2=macrophage chemoattractant protein MCP1 (3.3 fold)

Adrenomedullin ADM (2.8 fold in SKbr3 and 1.9 in BT474)

NFkappaB Pathway

a Interleukin 8(4.1 fold)

RelB (3.9 fold)

Sequestosome 1 (3.3 fold in SKbr3 and 2 fold in BT474)

Cell Adhesion

ICAM1 (4.3 fold)

Cadherin 5=Vascular endothelim (VE) cadherin (3.3 fold)

PLAUR=uPAR (3 fold)

DNA Damage Response

TIPARP (3 fold)

ATF3 (induced in both cell lines, stronger in BT474)

GADD45A

Oxidative Stress Response

- Pregnancy-induced growth inhibitor OKL38 (4.3 fold in SKBr3 and 2.3 fold in BT474)
- Glutamate cysteine ligase modifier subunit GCLM (about threefold in both SKBr3 and BT474)
- Heme oxygenase (11 fold in SKBrs)
- Cystathionine-beta-synthase (2.9 fold in SKBr3)

Other Groups

- Associated molecule with the SH3 domain of STAM (AMSH) kike (2.8 fold in SKBr3)

A micro array gene expression chip containing 225 unique genes, 112 up regulated and 113 down-regulated for the diagnosis of patients with adenocarcinoma to test eligibility for treatment with BZL101, an extract of Scutellaria barbata.

Tumors Expressing Genes, Up or Down Regulated, within the Common Paths Affected by BZL101 will be Eligible for BZL101 Treatment. These Tumors should Show Dependency or Damage in their Apoptosis, Cytokines with Proliferative Effects, NF Kappa B Pathway, Oxidative Stress, DNA Damage Response and Cell Adhesion Associated Genes.

TABLE 1

Genes up-reguated in response to BZL101

treatment in SKBr3 cells.

Functional
Entrez Gene
Average fold
Entrez

Entrez Gene Description
annotation
Name
upregulation
Gene ID

cytochrome P450, family 1,
Xenobiotic response
CYP1A1
43.9
1543

subfamily A, polypeptide 1

heme oxygenase
Oxidative
HMOX1
10.9
3162

(decycling) 1
response/NFkB

pathway

similar to Epigen protein

LOC255324
9.0
255324

chemokine (C—X—C motif)
NFkB pathway
CXCL1
7.3
2919

ligand 1 (melanoma growth

stimulating activity, alpha)

cytochrome P450, family 1,
Xenobiotic response
CYP1B1
6.8
1545

subfamily B, polypeptide 1

hypothetical gene

LOC440449
6.2
440449

supported by AF086204

tumor necrosis factor,
Oxidative/NFkB
TNFAIP3
6.1
7128

alpha-induced protein 3
pathway/Cell death

insulin growth factor-like
NFkB pathway
IGFL1
5.2
374918

family member 1

neuronal PAS domain

NPAS2
5.2
4862

protein 2

tumor necrosis factor (TNF
NFkB pathway/Cell
TNF
4.7
7124

superfamily, member 2)
death

cardiotrophin-like cytokine
NFkB pathway
CLC
4.6
23529

baculoviral IAP repeat-
NFkB pathway/Cell
BIRC3
4.6
330

containing 3
death

pregnancy-induced growth
Oxidative response
OKL38
4.3
29948

inhibitor

intercellular adhesion
NFkB pathway
ICAM1
4.3
3383

molecule 1 (CD54), human

rhinovirus receptor

interleukin 8
NFkB pathway
IL8
4.1
3576

v-rel reticuloendotheliosis
NFkB pathway
RELB
3.9
5971

viral oncogene homolog B,

nuclear factor of kappa

light polypeptide gene

enhancer in B-cells 3

(avian)

chemokine (C-C motif)
NFkB pathway/Cell
CCL2
3.4
6347

ligand 2
death

sequestosome 1
NFkB pathway/Cell
SQSTM1
3.3
8878

death

cardiotrophin-like cytokine
NFkB pathway/Cell
CLC
3.3
23529

death

cadherin 5, type 2, VE-

CDH5
3.3
1003

cadherin (vascular

epithelium)

TCDD-inducible
Oxidative/DNA
TIPARP
3.1
25976

poly(ADP-ribose)
damage

polymerase
response/NFkB

pathway

chemokine (C-C motif)
NFkB pathway
CCL2
3.0
6347

ligand 2

plasminogen activator,
NFkB pathway
PLAUR
3.0
5329

urokinase receptor

tumor necrosis factor
NFkB pathway/Cell
TNFRSF21
3.0
27242

receptor superfamily,
death

member 21

glutamate-cysteine ligase,
Oxidative
GCLM
2.9
2730

modifier subunit
response/NFkB

pathway

cystathionine-beta-synthase
Oxidative
CBS
2.9
875

response/NFkB

pathway

ring finger protein 24

RNF24
2.8
11237

associated molecule with

AMSH-LP
2.8
57559

the SH3 domain of STAM

(AMSH) like protein

adrenomedullin

ADM
2.8
133

heat shock 70 kDa protein 6
Xenobiotic response
HSPA6
2.8
3310

(HSP70B′)

BCL2 binding component 3
NFkB pathway/Cell
BBC3
2.7
27113

death

elongation factor, RNA

ELL2
2.6
22936

polymerase II, 2

activating transcription
Oxidative/DNA
ATF3
2.6
467

factor 3
damage

response/NFkB

pathway

chromosome 20 open

C20orf139
2.5
140809

reading frame 139

growth arrest and DNA-
DNA damage
GADD45A
2.5
1647

damage-inducible, alpha
response/Cell

death/NFkB pathway

spermidine/spermine N1-

SAT
2.5
6303

acetyltransferase

solute carrier organic anion

SLCO4A1
2.4
28231

transporter family, member

4A1

NK3 transcription factor

NKX3-1
2.4
4824

related, locus 1

(Drosophila)

chemokine (C-C motif)
NFkB pathway/Cell
CCL11
2.4
6356

ligand 11
death

maltase-glucoamylase

MGAM
2.3
8972

(alpha-glucosidase)

RAS, dexamethasone-
NFkB pathway
RASD1
2.3
51655

induced 1

zinc finger, SWIM domain

ZSWIM4
2.3
65249

containing 4

nuclear factor of kappa
NFkB pathway/Cell
NFKBIE
2.3
4794

light polypeptide gene
death

enhancer in B-cells

inhibitor, epsilon

p53-regulated apoptosis-
Cell death
P53AIP1
2.3
63970

inducing protein 1

NEDD9 interacting protein

NICAL
2.3
64780

with calponin homology

and LIM domains

CCR4 carbon catabolite

CCRN4L
2.3
25819

repression 4-like (S. cerevisiae)

Rap guanine nucleotide

RAPGEFL1
2.2
51195

exchange factor (GEF)-like 1

carbonic anhydrase VIII

CA8
2.2
767

arrestin domain containing 2

ARRDC2
2.2
27106

nasal embryonic LHRH

NELF
2.2
26012

factor

pleckstrin homology-like

PHLDA2
2.2
7262

domain, family A, member 2

claudin 1

CLDN1
2.2
9076

immediate early response 5

IER5
2.2
51278

phosphoinositide-3-kinase,

PIK3CD
2.2
5293

catalytic, delta polypeptide

pannexin 1

PANX1
2.2
24145

chemokine (C—X—C motif)
NFkB pathway/Cell
CXCL16
2.2
58191

ligand 16
death

keratin 5 (epidermolysis

KRT5
2.1
3852

bullosa simplex, Dowling-

Meara/Kobner/Weber-

Cockayne types)

myeloma overexpressed

MYEOV
2.1
26579

gene (in a subset of

t(11; 14) positive multiple

myelomas)

small nuclear RNA

SNAPC4
2.1
6621

activating complex,

polypeptide 4, 190 kDa

estrogen receptor binding

ERBP
2.1
30836

protein

ubiquitin-conjugating

UBE2E2
2.1
7325

enzyme E2E 2 (UBC4/5

homolog, yeast)

zinc finger protein 36, C3H

ZFP36
2.1
7538

type, homolog (mouse)

polo-like kinase 2

PLK2
2.1
10769

(Drosophila)

sulfide quinone reductase-

SQRDL
2.1
58472

like (yeast)

solute carrier family 2

SLC2A6
2.1
11182

(facilitated glucose

transporter), member 6

tubulin, beta 6

TUBB6
2.0
84617

RNA terminal phosphate

RCL1
2.0
10171

cyclase-like 1

ATP-binding cassette, sub-

ABCC2
2.0
1244

family C (CFTR/MRP),

member 2

leucine rich repeat and

LRFN1
2.0
57622

fibronectin type III domain

containing 1

pellino 3 alpha

MGC35521
2.0
246330

interferon regulatory factor 1
NFkB pathway
IRF1
2.0
3659

spermidine/spermine N1-

SAT
2.0
6303

acetyltransferase

optineurin

OPTN
2.0
10133

sestrin 2
Cell cycle regulation
SESN2
2.0
83667

tubulin, beta 2

TUBB2
2.0
7280

cytoplasmic

CPEB2
1.9
132864

polyadenylation element

binding protein 2

fatty acid desaturase 3

FADS3
1.9
3995

nuclear receptor coactivator 7

NCOA7
1.9
135112

TNF receptor-associated
NFkB pathway/Cell
TRAF3
1.9
7187

factor 3
death

tripartite motif-containing

TRIM21
1.9
6737

21

early growth response 1

EGR1
1.9
1958

MOB1, Mps One Binder

MOBKL2C
1.9
148932

kinase activator-like 2C

(yeast)

progressive external

PEO1
1.9
56652

ophthalmoplegia 1

DEAD (Asp-Glu-Ala-Asp)

DDX31
1.9
64794

box polypeptide 31

thioredoxin reductase 1
Oxidative response
TXNRD1
1.9
7296

endothelin 1
NFkB pathway
EDN1
1.9
1906

TIGA1

TIGA1
1.9
114915

transmembrane 4

TM4SF14
1.9
81619

superfamily member 14

axin 1

AXIN1
1.9
8312

monocyte to macrophage
Cell death
MMD
1.9
23531

differentiation-associated

CCAAT/enhancer binding

CEBPD
1.9
1052

protein (C/EBP), delta

leukotriene B4 receptor

LTB4R
1.9
1241

pre-B-cell colony
NFkB pathway
PBEF1
1.8
10135

enhancing factor 1

WD repeat domain 3

WDR3
1.8
10885

NIMA (never in mitosis
NFkB pathway
NEK6
1.8
10783

gene a)-related kinase 6

pleckstrin homology

PLEKHF1
1.8
79156

domain containing, family

F (with FYVE domain)

member 1

pleckstrin 2

PLEK2
1.8
26499

FKSG27 protein

FKSG27
1.8
126298

coronin, actin binding

CORO1C
1.8
23603

protein, 1C

snail homolog 1

SNAI1
1.8
6615

(Drosophila)

MOB1, Mps One Binder

MOBKL1A
1.8
92597

kinase activator-like 1A

(yeast)

cytochrome P450, family
Xenobiotic response
CYP27B1
1.8
1594

27, subfamily B,

polypeptide 1

pseudouridylate synthase 1

PUS1
1.8
80324

nuclear factor of kappa
NFkB pathway
NFKBIB
1.8
4793

light polypeptide gene

enhancer in B-cells

inhibitor, beta

ubiquitin-conjugating

UBE2E1
1.8
7324

enzyme E2E 1 (UBC4/5

homolog, yeast)

minor histocompatibility

HA-1
1.8
23526

antigen HA-1

PWP2 periodic tryptophan

PWP2H
1.7
5822

protein homolog (yeast)

tyrosylprotein

TPST1
1.7
8460

sulfotransferase 1

cyclin-dependent kinase
NFkB pathway/Cell
CDKN1A
1.7
1026

inhibitor 1A (p21, Cip1)
death

zinc finger protein 529

ZNF529
1.7
57711

TABLE 2

Genes down-reguated in response to

BZL101 treatment in SKBr3 cells.

Functional
Entrez
Ave Fold

Entrez Gene Description
annotataion
Gene Name
down regulation
Entrez_Gene_ID

inhibitor of DNA binding 1,
NFβB
ID1
5.0
3397

dominant negative helix-loop-
pathway/Cell

helix protein
cycle

regulation

cyclin-dependent kinase
Cell cycle
CDKN2C
3.3
1031

inhibitor 2C (p18, inhibits
regulation

CDK4)

zinc finger protein 339

ZNF339
2.9
58495

peroxiredoxin 3
Oxidative
PRDX3
2.9
10935

damage/NFβB

pathway

KIAA0644 gene product

KIAA0644
2.8
9865

eyes absent homolog 2

EYA2
2.6
2139

(Drosophila)

SMAD, mothers against DPP
Cell cycle
SMAD6
2.6
4091

homolog 6 (Drosophila)
regulation

ecotropic viral integration site 1

EVI1
2.6
2122

ras homolog gene family,

RHOU
2.6
58480

member U

gap junction protein, alpha 5,

GJA5
2.6
2702

40 kDa (connexin 40)

tensin

TNS
2.5
7145

RAB26, member RAS

RAB26
2.5
25837

oncogene family

chromosome 15 open reading

C15orf20
2.5
80119

frame 20

PTPRF interacting protein,

PPFIBP2
2.5
8495

binding protein 2 (liprin beta 2)

centrosomal protein 1

CEP1
2.5
11064

antigen identified by
Cell cycle
MKI67
2.4
4288

monoclonal antibody Ki-67
regulation

kinetochore associated 2
Cell cycle
KNTC2
2.4
10403

regulation

hyaluronan-mediated motility
Cell adhesion
HMMR
2.4
3161

receptor (RHAMM)

kinesin family member 20A

KIF20A
2.4
10112

zinc finger protein 467

ZNF467
2.4
168544

topoisomerase (DNA) II alpha
DNA damage
TOP2A
2.3
7153

170 kDa
response

inhibitor of DNA binding 2,
Cell cycle
ID2
2.3
3398

dominant negative helix-loop-
regulation

helix protein

MAX dimerization protein 3
Cell cycle
MXD3
2.3
83463

regulation

UDP-N-acetyl-alpha-D-

GALNT12
2.3
79695

galactosamine:polypeptide N-

acetylgalactosaminyltransferase

12 (GalNAc-T12)

transcription factor AP-2 beta

TFAP2B
2.3
7021

(activating enhancer binding

protein 2 beta)

centromere protein E, 312 kDa
Cell cycle
CENPE
2.3
1062

regulation

cell division cycle associated 3
Cell cycle
CDCA3
2.2
83461

regulation

sema domain, immunoglobulin

SEMA3F
2.2
6405

domain (Ig), short basic

domain, secreted, (semaphorin)

3F

myosin VIIA (Usher syndrome

MYO7A
2.2
4647

1B (autosomal recessive,

severe))

Rho-related BTB domain

RHOBTB3
2.2
22836

containing 3

otoraplin

OTOR
2.2
56914

centromere protein F,
Cell cycle
CENPF
2.2
1063

350/400ka (mitosin)
regulation

kinetochore protein Spc25
Cell cycle
Spc25
2.2
57405

regulation

CDC42 effector protein (Rho
Cell cycle
CDC42EP4
2.2
23580

GTPase binding) 4
regulation

baculoviral IAP repeat-
Cell cycle
BIRC5
2.2
332

containing 5 (survivin)
regulation/Cell

death

ankyrin 3, node of Ranvier

ANK3
2.2
288

(ankyrin G)

solute carrier family 40 (iron-

SLC40A1
2.2
30061

regulated transporter), member 1

peroxisomal biogenesis factor

PEX11A
2.2
8800

11A

Nedd4 binding protein 3

N4BP3
2.1
23138

SLIT-ROBO Rho GTPase

SRGAP2
2.1
23380

activating protein 2

cytochrome P450 4Z2

CYP4Z2P
2.1
163720

pseudogene

X-box binding protein 1
ER stress
XBP1
2.1
7494

ectodermal-neural cortex (with

ENC1
2.1
8507

BTB-like domain)

FGD1 family, member 3

FGD3
2.1
89846

centromere protein A, 17 kDa
Cell cycle
CENPA
2.1
1058

regulation

high-mobility group box 2
DNA damage
HMGB2
2.1
3148

response

dedicator of cytokinesis 11

DOCK11
2.1
139818

transducer of ERBB2, 1

TOB1
2.1
10140

sortilin-related receptor,

SORL1
2.1
6653

L(DLR class) A repeats-

containing

HRAS-like suppressor 3
Cell cycle
HRASLS3
2.1
11145

regulation

epithelial cell transforming
NFβB
ECT2
2.1
1894

sequence 2 oncogene
pathway

single-stranded DNA binding

SSBP2
2.1
23635

protein 2

nuclear factor I/A

NFIA
2.1
4774

delta sleep inducing peptide,

DSIPI
2.1
1831

immunoreactor

phosphodiesterase 8B

PDE8B
2.1
8622

BUB1 budding uninhibited by
Cell cycle
BUB1B
2.1
701

benzimidazoles 1 homolog beta
regulation

(yeast)

SWI/SNF related, matrix

SMARCA2
2.1
6595

associated, actin dependent

regulator of chromatin,

subfamily a, member 2

zinc finger protein 552

ZNF552
2.0
79818

polo-like kinase 1 (Drosophila)
Cell cycle
PLK1
2.0
5347

regulation

phosphoglucomutase 1

PGM1
2.0
5236

erythrocyte membrane protein

EPB41L1
2.0
2036

band 4.1-like 1

thioredoxin-related

TMX2
2.0
51075

transmembrane protein 2

kinesin family member 14

KIF14
2.0
9928

LPS-responsive vesicle

LRBA
2.0
987

trafficking, beach and anchor

containing

dehydrogenase/reductase (SDR

DHRS3
2.0
9249

family) member 3

pleckstrin homology domain

PLEKHK1
1.9
219790

containing, family K member 1

succinate-CoA ligase, GDP-

SUCLG1
1.9
8802

forming, alpha subunit

epithelial protein lost in

EPLIN
1.9
51474

neoplasm beta

histamine receptor H1

HRH1
1.9
3269

histone 1, H2ac

HIST1H2AC
1.9
8334

cingulin

CGN
1.9
57530

SAM and SH3 domain
Cell cycle
SASH1
1.9
23328

containing 1
regulation

tight junction protein 3 (zona

TJP3
1.9
27134

occludens 3)

mucin 15

MUC15
1.9
143662

chromosome condensation-
Cell cycle
CNAP1
1.9
9918

related SMC-associated protein 1
regulation

UDP-N-acetyl-alpha-D-

GALNT10
1.9
55568

galactosamine:polypeptide N-

acetylgalactosaminyltransferase

10 (GalNAc-T10)

aurora kinase B
Cell cycle
AURKB
1.9
9212

regulation

ATPase, H+ transporting,

ATP6V0A4
1.9
50617

lysosomal V0 subunit a

isoform 4

G-2 and S-phase expressed 1
DNA
GTSE1
1.9
51512

damage/Cell

cycle

regulation

spectrin repeat containing,

SYNE2
1.9
23224

nuclear envelope 2

heat shock 60 kDa protein 1

HSPD1
1.9
3329

(chaperonin)

nebulette

NEBL
1.9
10529

protein kinase C, delta

PRKCD
1.9
5580

neurexin 3
Cell adhesion
NRXN3
1.9
9369

transglutaminase 3 (E

TGM3
1.9
7053

polypeptide, protein-glutamine-

gamma-glutamyltransferase)

mesoderm posterior 1

MESP1
1.9
55897

hexosaminidase B (beta

HEXB
1.9
3074

polypeptide)

RWD domain containing 2

RWDD2
1.9
112611

fibroblast growth factor

FGFR2
1.9
2263

receptor 2 (bacteria-expressed

kinase, keratinocyte growth

factor receptor, craniofacial

dysostosis 1, Crouzon

syndrome, Pfeiffer syndrome,

Jackson-Weiss syndrome)

Down syndrome critical region

DSCR1
1.9
1827

gene 1

cyclin A2
DNA
CCNA2
1.8
890

damage/Cell

cycle

regulation

calcyphosine

CAPS
1.8
828

methylcrotonoyl-Coenzyme A

MCCC1
1.8
56922

carboxylase 1 (alpha)

keratin 15

KRT15
1.8
3866

GPAA1P anchor attachment

GPAA1
1.8
8733

protein 1 homolog (yeast)

actin related protein 2/3
Cell adhesion
ARPC5
1.8
10092

complex, subunit 5, 16 kDa

ATP-binding cassette, sub-

ABCA1
1.8
19

family A (ABC1), member 1

epidermal growth factor

EPS8
1.8
2059

receptor pathway substrate 8

HMBA-inducible
Cell cycle
HIS1
1.8
10614

regulation

transcription elongation factor

TCEAL1
1.8
9338

A (SII)-like 1

breast carcinoma amplified

BCAS1
1.8
8537

sequence 1

distal-less homeobox 4

DLX4
1.8
1748

protein phosphatase 1H (PP2C

PPM1H
1.8
57460

domain containing)

heat shock 70 kDa protein 8

HSPA8
1.8
3312

Kruppel-like factor 13

KLF13
1.8
51621

15 kDa selenoprotein

15-$$
1.7
9403

eukaryotic translation initiation

EIF4EBP2
1.7
1979

factor 4E binding protein 2

branched chain keto acid

BCKDHA
1.7
593

dehydrogenase E1, alpha

polypeptide (maple syrup urine

disease)

eukaryotic translation initiation

EIF3S6IP
1.7
51386

factor 3, subunit 6 interacting

protein

zinc fingers and homeoboxes 2

ZHX2
1.7
22882

SUMO-1 activating enzyme

SAE1
1.7
10055

subunit 1

Example 3
Methods of Diagnosing and Treating Humans with Breast Cancer Using 8-oxoguanine Biomarker

A group of up to thirty patients who have and who do not have breast cancer will be tested for the presence of 8-oxoguanine. All patients will be at least 18 years of age. Patients representing an individual having breast cancer will present both histological confirmation of breast cancer as well as clinical evidence of metastatic involvement. The testing will be done in a double blind fashion. Patients who enter into the test will have a blood sample drawn. The blood will be tested for presence of 8-oxoguanine biomarker by reacting the blood with a diagnostic tool that detects the presence of 8-oxoguanine. For each patient, a level of 8-oxoguanine will be output to a display indicating the level of 8-oxoguanine for that specific patient. The patient's level of 8-oxoguanine will then be compared to a predetermined level of 8-oxoguanine as determined from sampling a population of 50 individuals who have not been diagnosed with breast cancer, or normal individuals. The predetermined threshold level will include the average 8-oxoguanine level for the group of normal individuals plus and minus the standard deviation for the population. Patients in whom a level of 8-oxoguanine biomarker is detected to be above the threshold level will then be singled out as potentially having breast cancer. These patients will then be compared to any histological data present for that patient. Once it is confirmed that the patient does in fact have breast cancer, the patient will be administered 350 ml (equivalent to 12 grams dry soluble extract of 180 grams of BZL) BZL101 extract per day. These patients will then be monitored by having blood drawn at specified intervals, preferably once a month to detect levels of 8-oxoguanine in the blood. BZL101 will continue to be administered to the patient until an adverse event more severe than a grade I or grade II adverse event occurs, the patient self-elects to forego treatment or the disease progresses to a clinically significant degree. Throughout treatment, periodically (e.g. once per week), the level of 8-oxoguanine biomarker will be detected to track the progress of treatment.

Example 3
Methods of Diagnosing and Treating Humans with Breast Cancer Using Lactate Dehydrogenase and a 8-oxoguanine Biomarkers

A group of thirty patients who have and who do not have breast cancer will be tested for the presence of lactate dehydrogenase and 8-oxoguanine biomarkers. All patients were at least 18 years of age. Patients representing an individual having breast cancer will present both histological confirmation of breast cancer as well as clinical evidence of metastatic involvement. The testing will be done in a double blind fashion. Patients who enter into the test will submit a urine sample. The urine sample will be tested for presence of lactate dehydrogenase and 8-oxoguanine biomarkers by reacting the urine with a diagnostic tool that detects the presence of both the lactate dehydrogenase and the 8-oxoguanine biomarkers. For each patient, a level of lactate dehydrogenase and 8-oxoguanine will be output to a display indicating the levels of lactate dehydrogenase and 8-oxoguanine for that specific patient. The patient's level of lactate dehydrogenase and 8-oxoguanine will then be compared to a predetermined levels of lactate dehydrogenase and 8-oxoguanine as determined from sampling a population of 50 individuals who have not been diagnosed with breast cancer, or normal individuals. The predetermined threshold levels will include the average lactate dehydrogenase and 8-oxoguanine levels for the group of normal individuals plus and minus the standard deviation for the population. Patients in whom a level of lactate dehydrogenase and 8-oxoguanine biomarker is detected to be above the threshold level will then be singled out as potentially having breast cancer. These patients will then be compared to any histological data present for that patient Once it is confirmed that the patient does in fact have breast cancer, the patient will be administered a therapeutic amount of an extract of BZL101 in order to treat the cancer.

CONCLUSION

The herbal extract BZL101, its uses for the inhibition of solid tumor cancer cells and the treatment of such cancers in patients are described herein. Although certain embodiments and examples have been used to describe the present invention, it will be apparent to those skilled in the art that changes to the embodiments and examples may be made without departing from the scope and spirit of this invention.

While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

METHODS OF DETECTING AND TREATMENT OF CANCERS USING SCUTELLARIA BARBATA EXTRACT

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