JP2012521424A - Methods and compositions for the treatment of cancer - Google Patents

Abstract

An isolated compound and a combination of isolated compounds isolated from Streptomyces are effective in generating reactive oxygen species, inducing DNA damage and inducing apoptosis in cancer cells. The compounds and combinations can be prepared as pharmaceutical compositions for administration to mammals, such as humans, for the treatment of solid cancers, such as epithelial cancers. Such epithelial cancers include breast cancer and ovarian cancer.
[Selection] Figure 1

Description

Cross reference
[0001] This application is from US patent application Ser. No. 12 / 553,878, filed Sep. 3, 2009, and US Provisional Application No. 61 / 162,988, filed Mar. 24, 2009. And the benefit of priority from US Provisional Application No. 61 / 172,639, filed April 24, 2009, each of which is incorporated herein by reference in its entirety.

  [0002] Advances in early detection and combination therapy for breast cancer generally have promising effects on patient survival, but patients with advanced metastatic breast cancer generally face a less prognostic prognosis It is. Commonly used hormonal and chemotherapeutic drugs can lead to temporary regression of tumors and can relieve symptoms associated with cancer. However, these treatments are often accompanied by toxicity and intolerable side effects and eventually become ineffective in controlling advanced breast cancer and its symptoms. The improvement in breast cancer survival is not very great even with newer targeted biological agents. Moreover, in most metastatic cancers, resistance to commonly available commonly used treatments eventually develops or patients experience excessive side effects.

  [0003] It is interesting to note that more 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 tree of Pacific yew, Taxus brevifolia. Around the world, it is estimated that approximately 80% of the world population still relies on botanical drugs as the main source of therapy. In the West, botanical drugs are considered a common form of supplemental and alternative drugs among patients diagnosed with cancer. However, despite the anecdotal case reports of curative and clinical efficacy in women who have relied solely on botanical drugs for treatment, firmly evaluate the safety and efficacy of botanical drugs for the treatment of breast cancer There have been few clinical trials. Previously, it has been shown that aqueous extracts of Scutellaria barbata can provide growth inhibition of breast cancer cell lines in vitro (“In vitro antiproliferative activity of Chinese medicinal herbs against breast cancer cells”) Anticancer Res., 22 (6C): 3843-52 (2002)). BZL101, an enriched aqueous extract of half-branched lotus, was evaluated for anti-proliferative activity against five breast cancer cell lines (SK-BR-3, MCF7, MDA-MB-231, BT-474, and MCNeuA) . These cell lines represent an important prognostic phenotype of breast cancer that expresses a range of estrogen and HER2 receptors. BZL101 tested at a 1:10 dilution (15 μg / ml) showed over 50% growth inhibition against 4 of its 5 cell lines (Campbell, 2002). BZL101 showed over 50% growth inhibition against a group of lung, prostate and pancreatic cancer cell lines. BZL101 at the same dose did not cause more than 25% growth inhibition over normal human breast cells (HuMEC), indicating selectivity for cancer cells (Table 1). Furthermore, BZL101 had a mild mitogenic effect on normal human lymphocytes. In cell cycle analysis, BZL101 caused S phase burst and G1 arrest. BZL101 also weakened the mitochondrial membrane potential and caused caspase-independent polymer grade (HMG) apoptosis.

Newman 2003 Anticancer Res., 22 (6C): 3843-52 (2002) Campbell, 2002

  [0004] There is a need for therapies for the treatment of patients with metastatic cancer. There is also a need for a therapy that has reduced, more specifically minimal toxicity, for patients with metastatic cancer. In particular, there is a need for new therapies with relatively low toxicity for the treatment of metastatic solid tumors such as epithelial tumors, and more particularly breast and ovarian cancer.

[0005] These and other needs are met by aspects of the present invention.
Summary of the Invention
[0006] The inventor has found that extracts of Scutellaria barbata D. Don are well tolerated at much higher doses than previously reported. The extract of Selexanthus is well tolerated at a dose of soluble material extracted from at least about 20 g of Selexanthus. Furthermore, the inventor has found that an extract of Selexanthus can be conveniently provided in a dosage unit suitable for administration to a patient. Accordingly, in some embodiments, a dosage unit comprising at least about 20 g of soluble material extracted from Selexanthus is provided. In some embodiments, the unit dose further comprises at least one excipient, in particular at least one excipient other than water, and in particular at least one flavoring agent, sweetener or both. In certain embodiments, the dosage unit is in a form suitable for oral administration, such as an aqueous (water-based) composition or a dry powder suitable for reconstitution with water. The inventor has found that the dosage unit is suitable for administration to cancer patients, particularly cancer patients suffering from breast cancer or gynecological cancers such as uterine cancer.

  [0007] The inventor has determined that a dose of soluble material extracted from at least about 20 g of Selexanthus per day is well tolerated and effective in the treatment of cancer, particularly breast cancer. Accordingly, the present invention provides a method of treating cancer comprising administering to a cancer patient at least about 20 g of soluble material extracted from Selexanthus per day. In some embodiments, the cancer is selected from breast cancer and one or more gynecological cancers. In some embodiments, the method comprises administering to the patient about 20 g / day to about 200 g / day of soluble material extracted from Sedum.

  [0008] The inventor has also determined that the addition of excipients, such as a corrigent, to a pharmaceutical composition containing a high dose of an extract of Selexanthus can attenuate the bitter taste of the extract. Since the inventor has found that high doses of Selexanthus pertussis (eg, at least about 20 g of soluble substance per administration or per day) are well tolerated but are relatively unpalatable, the inventor It has been found that the addition of a flavoring agent or other agent is desirable to make it palatable, for example, for high dose consumption of the extract of Selexanthus extract for the treatment of cancer. Accordingly, embodiments described herein include at least one excipient other than water (eg, at least one flavoring agent, sweetener or both), as well as the anticancer activity of the inventor. Contains one or more members of the group consisting of luteolin, apigenin, scutellarein, and scutellarin, which are anti-cancer active substances identified as being necessary (eg, cancer, particularly breast cancer) Or a pharmaceutical composition (for the treatment of gynecological cancer). The inventor has found that high molecular weight compounds, such as compounds having a high molecular weight (eg, a molecular weight greater than 1,000 to 10,000 grams / mole) increase the bulk of the composition without imparting any substantial value activity, It has also been found to tend to cause stomach upset, gas, abdominal distension and / or diarrhea. Accordingly, in some embodiments, the pharmaceutical composition is depleted of high molecular weight compounds and in some embodiments is substantially free of high molecular weight compounds. In some embodiments, the composition comprises about 1 part luteolin, about 1.1 parts apigenin, about 4.8 parts scutellarein and about 34 parts scutellarin, comprising luteolin, apigenin, scutellarein, and scutellarine. Includes combinations. (All “parts” are determined by weight.) In some embodiments, the combination of luteolin, apigenin, scutellarein, and scutellarin is about 1 part luteolin, about 0.61 to about 2 parts apigenin, about 2.5 to about 9.4 parts of scutellarein, and about 15 to about 70 parts of scutellarin. In some embodiments, the composition comprises about 1 g to about 200 g of soluble material, about 1% to about 99% of the soluble material is active soluble material, and about 1.7% to about 1.7% of the effective soluble material. About 3.2% is luteolin, about 2% to about 3.4% is apigenin, about 7.9% to about 15.8% is scutellarein, about 49% to the rest of the effective soluble material The part is scutellarin. In some embodiments, the composition is selected from one or more of advanced breast cancer, metastatic breast cancer, treatment resistant breast cancer, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, and / or triple negative breast cancer. Used to treat breast cancer.

  [0009] Some embodiments described herein are methods of treating cancer, particularly one or more breast and / or gynecological cancers, wherein the patient has an effective amount of at least one water. Providing a composition comprising administering a non-excipient excipient (eg, at least one flavoring agent, sweetener or both) and one or more members of the group consisting of luteolin, apigenin, scutellarein, and scutellarin To do. In some embodiments, the composition is selected from one or more of advanced breast cancer, metastatic breast cancer, treatment resistant breast cancer, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, and / or triple negative breast cancer. Used to treat breast cancer. In some embodiments, the effective amount of the composition comprises a combination of at least 0.25 g luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the effective amount of the composition comprises a combination of at least 0.27 g luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the composition comprises a combination of at least 0.35 g luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the effective amount of the composition is about 0.35 g to 2 g, about 0.35 g to 1.1 g, about 0.35 g to 1 g, about 0.35 g to about 0.8 g luteolin, apigenin, Includes a combination of scutellarein and scutellarin.

  [0010] The inventor has found that the combination of luteolin, apigenin, scutellarein, and scutellarin is effective as a treatment for cancer, particularly breast cancer. Accordingly, in some embodiments, the present invention provides a dosage unit comprising a combination of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the dosage unit comprises at least about 0.25 g, at least about 0.27 g, or at least about 0.35 g of a combination of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the dosage unit further comprises at least one excipient other than water, such as a flavoring agent, sweetener, or both. In some embodiments, the dosage unit is substantially free of high molecular weight compounds extracted from Sedum. In some embodiments, the composition is used in a method of treating cancer, such as breast cancer and / or one or more gynecological cancers. In some embodiments, the cancer is breast cancer, such as advanced breast cancer, metastatic breast cancer, breast cancer resistant to treatment, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, and / or triple negative breast cancer.

  [0011] The inventor further discovered a process for making a pharmaceutical composition using as part of the starting material an aerial portion of Selexanthus. Such a process is particularly useful for making compositions comprising luteolin, apigenin, scutellarein, and scutellarin. Accordingly, in some embodiments, the inventor has described a process for making a pharmaceutical composition that includes: (a) water of a portion of the sorghum that grows in air above 40 ° C. Contacting for a period of at least about 10 minutes to form a mixture; (b) separating the portion of the sedum that grows in air to form a crude extract; (c) the crude extraction Separating high molecular weight compounds from the product to form a purified extract; (d) optionally evaporating some, substantially all, or all of the water from the purified extract, or purification thereof Additional water is added to the extracted extract; and (e) the purified extract is combined with at least one excipient other than pharmaceutically acceptable water to form the pharmaceutical composition. In some embodiments, the purified extract comprises apigenin, luteolin, scutellarein, and scutellarin. In some embodiments, the at least one pharmaceutically acceptable non-water excipient is selected from flavoring agents and sweeteners.

  [0012] The inventor has found that removal of at least a part of the high molecular weight compound extracted from Sedum is improving the clinical characteristics of the pharmaceutical composition. Since a large amount of soluble substances extracted from St. elegans is inactive, reducing the amount of soluble substances by removing molecules with a molecular weight above a predetermined cut-off value is The bulk of the pharmaceutical composition derived from the product will be greatly reduced. In addition, most of the soluble material extracted into water from Selexanthus is soluble fiber, which is not absorbed in the intestine and tends to promote gastrointestinal upset, abdominal distension, gas and diarrhea. Thus, removing at least a portion of its soluble fiber by reducing the burden of soluble material extracted from Selexanthus, while retaining a mixture of luteolin, apigenin, scutellarein, and scutellarin in the composition Resulting in improved anticancer drugs. Thus, in some embodiments, the invention provides a pharmaceutical composition comprising 1 part of a combination of luteolin, apigenin, scutellarein, and scutellarin and a high molecular weight compound having a molecular weight greater than a predetermined cut-off value of less than about 50 parts. Wherein the predetermined cutoff value is from 1,000 grams / mole to about 20,000 grams / mole.

  [0013] The inventor has also discovered a process for making a pharmaceutical composition that includes: (a) a portion of an A. pertussis grown in air with water heated to above 40 ° C. and a period of at least about 10 minutes To form a mixture; and (b) separating the portion of the cereals that grew in the air from the mixture to produce a crude extract; (c) producing a high molecular weight compound from the crude extract; Separating to form a purified extract; (d) optionally evaporating some, substantially all, or all of the water from the purified extract or adding to the purified extract And (e) combining the purified extract with a pharmaceutically acceptable excipient to form the pharmaceutical composition. 1 part luteolin, about 0.61 to about 2 parts apigenin, about 2.5 to about 9.4 parts scutellarein, and about 15 to about 70 parts scutellarin.

  [0014] Some embodiments also provide a process for making a purified extract of St. abyss, comprising: (a) heating a portion of St. abyss grown in air to above 40 ° C Contacting with water for a period of at least about 10 minutes to form a mixture; (b) separating the portion of the cereals that grew in the air from the mixture to produce a crude extract; and (c) The high molecular weight compound is separated from the crude extract to form a purified extract of Selexanthus.

  [0015] Some embodiments are further processes for making a pharmaceutical composition, wherein at least one pharmaceutically acceptable excipient other than water is one of the group consisting of luteolin, apigenin, scutellarein, and scutellarin A process comprising combining the above members to form a pharmaceutical composition thereof is provided. In some embodiments, the at least one non-water pharmaceutical excipient is selected from flavoring agents and sweeteners.

  [0016] Some embodiments provide a process for making a pharmaceutical dosage unit comprising: (a) at least a portion of water that is grown in the air of Selexanthus and heated to above 40 ° C and at least Forming a mixture upon contact for a period of about 10 minutes; (b) separating the portion of the cedar mushrooms grown in air to form a crude extract; and (c) the crude extraction Separating the high molecular weight compound from the product to form a purified extract; and (d) combining the purified extract with at least one excipient other than water to form the pharmaceutical dosage unit. To do. In some embodiments, the at least one non-water excipient is selected from flavoring agents and sweeteners.

[0017] Other uses and advantages of the invention will become apparent to those skilled in the art after considering the description, including the drawings and claims herein.
Support
[0018] All publications and patent applications mentioned in this specification are herein incorporated to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated. Incorporated into.

  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:

[0020] FIG. 1 shows the effect of various active compounds extracted from Solidago on the production of reactive oxygen species (ROS) as measured by DCFDA fluorescence. [0021] FIG. 2 shows the effect on active oxygen species (ROS) production of various active compounds extracted from Sedum, measured by dihydroethidium (HE) fluorescence. [0022] FIG. 3 shows the effect of various active compounds extracted from Sedum on the production of mitochondrial reactive oxygen species (ROS) as measured by MitoSOX fluorescence. [0023] FIG. 4 shows the effect of various active compounds extracted from Solidago on the production of comets in treated cells. [0024] FIG. 5 shows the effect of various active compounds extracted from Sedum on the ATP production in the treated cells.

  [0025] The present invention relates to pharmaceutical compositions and active dosages comprising an active agent isolated from a branch lotus extract, and to methods of using these extracts for the treatment of cancer. In a particular embodiment, the herb from which the active compound is separated is selected from the species of Lamiaceae.

  [0026] In addition, the present invention relates to a method of using an extract of Selexus pertussis, whereby the extract of Selexanthus is administered to a patient at a dose not previously characterized.

  [0027] The present invention further relates to the administration of an effective drug and an effective drug combination derived from an extract of Selexus mirabilis, an extract of Selexus mirabilis, and in particular an aqueous extract of Selexus mirabilis.

  [0028] The inventor has found that extracts of Selexanthus are well tolerated even at doses much higher than previously reported, eg, at least about 20 g / day of soluble material extracted from Selexanthus It has been found that it can be administered to patients without inducing limiting toxicity. The inventor administered 20 g / day, 30 g / day, and 40 g / day of a soluble substance extracted from St. auricularia to breast cancer patients without reaching the maximum tolerated capacity. As such, the inventors have determined that doses of at least about 20 g / day, particularly from about 20 g / day to about 200 g / day, are within the scope of the present invention. Furthermore, the inventor has found that an extract of Selexanthus can be conveniently provided in a dosage unit suitable for administration to a patient. Accordingly, in some embodiments, a dosage unit comprising at least about 20 g of soluble material extracted from Selexanthus is provided. In some embodiments, the unit dose further comprises at least one excipient, in particular at least one excipient other than water, and in particular at least one flavoring agent, sweetener or both. In certain embodiments, the dosage unit is in a form suitable for oral administration, such as an aqueous (water-based) composition or a dry powder suitable for reconstitution with water. The inventor has found that the dosage unit is suitable for administration to cancer patients, particularly cancer patients suffering from breast cancer or gynecological cancers such as uterine cancer. In certain embodiments, the dosage unit comprises from about 20 g to about 200 g of soluble material extracted from Sedum. In some embodiments, the dosage unit is about 20 g to 100 g, about 20 g to 60 g, about 20 g to 50 g, about 20 g to 40 g, about 20 g, about 30 g, about 40 g, about 50 g, about 60 g, or about 40 g to Contains soluble material extracted from 100 g of Selex. The inventor has also identified the anticancer active compounds of Selexiana (ie, luteolin, apigenin, scutellarein, and scutellarin) and their concentrations in soluble substances extracted from Selexus, which is at least about 0.25 g. A dosage unit of the combination of luteolin, apigenin, scutellarein, and scutellarin is also provided. Some embodiments have at least about 0.27 g, at least about 0.35 g, about 0.35 g to 4 g, about 0.35 g to 2 g, about 0.35 g to 1.1 g, about 0.35 g to about 1 g, or A dosage unit of about 0.35 g to about 0.8 g of a combination of luteolin, apigenin, scutellarein, and scutellarin is provided.

  [0029] The inventor has determined that a dose of soluble material extracted from at least about 20 g of Selexanthus per day is well tolerated and effective in the treatment of cancer; Provided is a method of treating cancer comprising administering to a cancer patient at least about 20 g of soluble material extracted from Selexanthus per day. In some embodiments, the cancer is selected from breast cancer and one or more gynecological cancers. In some embodiments, the cancer is breast cancer. In some embodiments, the breast cancer is advanced breast cancer, metastatic breast cancer, treatment resistant breast cancer, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, and / or triple negative breast cancer. In some embodiments, the method comprises administering to the patient about 20 g / day to about 200 g / day of soluble material extracted from Sedum. In some embodiments, the patient has about 20 g per day to 100 g per day, about 20 g per day to 60 g per day, about 20 g per day to 50 g per day, about 20 g per day 40 g per day, or about 40 g per day to 100 g per day, of soluble material extracted from Sedum. In some embodiments, the soluble material extracted from the scabbard contains at least about 0.25 g of a combination of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the soluble material extracted from the scabbard contains at least about 0.27 g of a combination of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the soluble material extracted from the scabbard contains at least about 0.35 g of a combination of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the soluble material extracted from the Selexanthus is about 0.35 g to 4 g, about 0.35 g to 2 g, about 0.35 g to 1.1 g, about 0.35 g to 1 g, about 0.35 g. -0.8g, about 0.35g-0.75g, or about 0.7g-2g of a combination of luteolin, apigenin, scutellarein, and scutellarin.

  [0030] The inventor has also determined that the addition of excipients, such as a corrigent, to a pharmaceutical composition containing a high dose of an extract of Selexanthus can attenuate the bitter taste of the extract. Since the inventor has found that high doses of Selexanthus pertussis (eg, at least about 1 g of soluble material per administration or per day) are well tolerated but relatively bad taste, the inventor It has been found that the addition of a flavoring agent or other agent is desirable to make it palatable due to the high dose consumption of, for example, Selexanthus extract for the treatment of cancer. Accordingly, embodiments described herein include anticancer activity of at least one excipient other than water (eg, at least one flavoring agent, sweetener, or both), as well as an extract by the inventor. Including one or more members of the group consisting of luteolin, apigenin, scutellarein, and scutellarin, which are anti-cancer active substances identified as being necessary for cancer (eg, for the treatment of cancer, particularly breast cancer or gynecological cancer) A pharmaceutical composition is provided. The inventor believes that high molecular weight compounds, such as compounds having a molecular weight greater than 1000 g / mol (although other cut-off values may be used, eg 1,000 to 10,000 g / mol) are of some substantial value. It has also been found that the composition tends to increase in bulk without imparting activity, causing stomach upset, abdominal distension and / or diarrhea. Accordingly, in some embodiments, the pharmaceutical composition is depleted of high molecular weight compounds and in some embodiments is substantially free of high molecular weight compounds. In some embodiments, the composition comprises about 1 part luteolin, about 1.1 parts apigenin, about 4.8 parts scutellarein and about 34 parts scutellarin, comprising luteolin, apigenin, scutellarein, and scutellarine. Includes combinations. (All “parts” are determined by weight.) In some embodiments, the combination of luteolin, apigenin, scutellarein, and scutellarin is about 1 part luteolin, about 0.9 to about 1.3 parts apigenin. About 3.9 to about 6 parts scutellarein, and about 37 to about 43 parts scutellarin. In some embodiments, the combination of luteolin, apigenin, scutellarein, and scutellarin is about 1 part luteolin, about 0.75 to about 1.64 parts apigenin, about 3.1 to about 7.5 parts scutellarein, And about 20.4 to about 54.7 parts scutellarin. In some embodiments, the combination of luteolin, apigenin, scutellarein, and scutellarin is about 1 part luteolin, about 0.61 to about 2 parts apigenin, about 2.5 to about 9.4 parts scutellarein, and about Contains 15 to about 70 parts scutellarin. In some embodiments, the composition comprises about 1 g to about 200 g of soluble material, about 1% to about 99% of the soluble material is active soluble material, and about 1.7% to about 1.7% of the effective soluble material. About 3.2% is luteolin, about 2% to about 3.4% is apigenin, about 7.9% to about 15.8% is scutellarein, about 49% to the rest of the effective soluble material The part is scutellarin. In some embodiments, the composition comprises about 1 g to about 200 g of soluble material, about 1% to about 3% of the soluble material is active soluble material, and about 1.7% to about 1.7% of the effective soluble material. About 3.2% is luteolin, about 2% to about 3.4% is apigenin, about 7.9% to about 15.8% is scutellarein, about 49% to the rest of the effective soluble material The part is scutellarin. In some embodiments, the composition comprises about 1 g to about 200 g of soluble material, about 1.5% to about 2.1% of the soluble material is active soluble material, and about 1 of the effective soluble material. 0.7% to about 3.2% is luteolin, about 2% to about 3.4% is apigenin, about 7.9% to about 15.8% is scutellarein, about 49% to effective solubility The rest of the material is scutellarin. In some embodiments, the composition comprises about 1 g to about 200 g of soluble material, about 1% to about 99% of the soluble material is active soluble material, and about 1.9% to about 1.9% of the effective soluble material. About 3% is luteolin, about 2.2% to about 3.2% is apigenin, about 9.2% to about 14.5% is scutellarein, about 60% to the remainder of the effective soluble material The part is scutellarin. In some embodiments, the composition comprises about 1 g to about 200 g of soluble material, about 1% to about 3% of the soluble material is active soluble material, and about 1.9% to about 1.9% of the effective soluble material. About 3% is luteolin, about 2.2% to about 3.2% is apigenin, about 9.2% to about 14.5% is scutellarein, about 60% to the remainder of the effective soluble material The part is scutellarin. In some embodiments, the composition comprises about 1 g to about 200 g of soluble material, about 1.5% to about 2.1% of the soluble material is active soluble material, and about 1 of the effective soluble material. .9% to about 3% is luteolin, about 2.2% to about 3.2% is apigenin, about 9.2% to about 14.5% is scutellarein, about 60% to effective solubility The rest of the material is scutellarin. In some embodiments, the composition comprises from about 1 g to about 200 g of soluble material, from about 1% to about 50% of the soluble material is effective soluble material, from about 2.2% of the effective soluble material About 2.7% is luteolin, about 2.4% to about 2.9% is apigenin, about 10.5% to about 13.2% is scutellarein, about 72% to an effective soluble substance The rest is scutellarin. In some embodiments, the composition comprises from about 1 g to about 200 g of soluble material, from about 1% to about 3% of the soluble material is effective soluble material, from about 2.2% of the effective soluble material About 2.7% is luteolin, about 2.4% to about 2.9% is apigenin, about 10.5% to about 13.2% is scutellarein, about 72% to an effective soluble substance The rest is scutellarin. In some embodiments, the composition comprises about 1 g to about 200 g of soluble material, about 1.5% to about 2.1% of the soluble material is active soluble material, and about 2 of the effective soluble material. 2% to about 2.7% is luteolin, about 2.4% to about 2.9% is apigenin, about 10.5% to about 13.2% is scutellarein, about 72% to The remaining portion of the effective soluble material is scutellarin. In some embodiments, the composition is selected from one or more of advanced breast cancer, metastatic breast cancer, treatment resistant breast cancer, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, and / or triple negative breast cancer. Used to treat breast cancer.

  [0031] Some embodiments described herein are methods of treating cancer, particularly one or more breast and / or gynecological cancers, wherein the patient has an effective amount of at least one Administering a composition comprising an excipient other than water (eg, at least one flavoring agent, sweetener or both) and one or more members of the group consisting of luteolin, apigenin, scutellarein, and scutellarin. provide. In some embodiments, the composition is selected from one or more of advanced breast cancer, metastatic breast cancer, treatment resistant breast cancer, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, and / or triple negative breast cancer. Used to treat breast cancer. In some embodiments, the effective amount of the composition comprises a combination of at least 0.25 g luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the effective amount of the composition comprises a combination of at least 0.27 g luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the composition comprises a combination of at least 0.35 g luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the effective amount of the composition comprises from about 0.27 g to about 4 g of a combination of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the effective amount of the composition comprises from about 0.35 g to about 4 g of a combination of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the effective amount of the composition is about 0.35 g to 2 g, about 0.35 g to 1.1 g, about 0.35 g to 1 g, about 0.35 g to about 0.8 g luteolin, apigenin, Includes a combination of scutellarein and scutellarin. In some embodiments, the combination of luteolin, apigenin, scutellarein, and scutellarin includes: about 1 part luteolin, about 1.1 parts apigenin, about 4.8 parts scutellarein, and about 34 parts About 1 part luteolin, about 0.9 to about 1.3 parts apigenin, about 3.9 to about 6 parts scutellarein, and about 37 to about 43 parts scutellarin; about 1 part luteolin, about 0 .75 to about 1.64 parts apigenin, about 3.1 to about 7.5 parts scutellarein, and about 20.4 to about 54.7 parts scutellarin; about 1 part luteolin, about 0.61 to about 2 parts apigenin, about 2.5 to about 9.4 parts scutellarein, and about 15 to about 70 parts scutellarin; about 6.7 mg to about 90 mg luteolin, about 8.9 mg to about 9 about 8.9 mg to about 50 mg luteolin, about 8.9 mg to about 30 mg luteolin, about 8.9 mg to about 25 mg luteolin, or about 8.9 mg to about 20 mg luteolin; About 100 mg apigenin, about 9.7 mg to about 100 mg apigenin, about 9.7 mg to about 50 mg apigenin, about 9.7 mg to about 30 mg apigenin, about 9.7 mg to about 25 mg apigenin, or about 9.7 mg To about 20 mg apigenin; about 30 mg to about 500 mg scutellarein, about 40 mg to about 500 mg scutellarein, about 40 mg to about 220 mg scutellarein, about 40 mg to about 130 mg scutellarein, about 40 mg to about 110 mg scutellarein, or about 40 mg to About 90 mg of scutellare About 0.25 g to about 3 g of scutellarin, about 0.3 g to about 3 g of scutellarin, about 0.3 g to about 1.5 g of scutellarin, about 0.3 g to about 0.9 g of scutellarin, about 0.3 g ~ About 0.8 g of scutellarin, or about 0.3 g to about 0.65 g of scutellarin.

  [0032] The inventor has found that the combination of luteolin, apigenin, scutellarein, and scutellarin is effective in the treatment of cancer, particularly breast cancer. Accordingly, in some embodiments, the present invention provides a dosage unit comprising a combination of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the dosage unit comprises a combination of at least about 0.25 g luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the dosage unit comprises at least about 0.27 g, or at least about 0.35 g of a combination of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the dosage unit is from about 0.35 g to about 4 g, from about 0.35 g to about 2 g, from about 0.35 g to about 1.1 g, from about 0.35 g to about 1 g, from about 0.35 g to Contains about 0.8 g, or about 0.7 g to about 2 g of a combination of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the dosage unit further comprises at least one excipient other than water, such as a flavoring agent, sweetener, or both. In some embodiments, the dosage unit is substantially free of high molecular weight compounds extracted from Sedum. In some embodiments, the combination of luteolin, apigenin, scutellarein, and scutellarin includes: about 1 part luteolin, about 1.1 parts apigenin, about 4.8 parts scutellarein, and about 34 parts About 1 part luteolin, about 0.9 to about 1.3 parts apigenin, about 3.9 to about 6 parts scutellarein, and about 37 to about 43 parts scutellarin; about 1 part luteolin, about 0 .75 to about 1.64 parts apigenin, about 3.1 to about 7.5 parts scutellarein, and about 20.4 to about 54.7 parts scutellarin; about 1 part luteolin, about 0.61 to about 2 parts apigenin, about 2.5 to about 9.4 parts scutellarein, and about 15 to about 70 parts scutellarin; about 6.7 mg to about 90 mg luteolin, about 8.9 mg to about 9 about 8.9 mg to about 50 mg luteolin, about 8.9 mg to about 30 mg luteolin, about 8.9 mg to about 25 mg luteolin, or about 8.9 mg to about 20 mg luteolin; About 100 mg apigenin, about 9.7 mg to about 100 mg apigenin, about 9.7 mg to about 50 mg apigenin, about 9.7 mg to about 30 mg apigenin, about 9.7 mg to about 25 mg apigenin, or about 9.7 mg To about 20 mg apigenin; about 30 mg to about 500 mg scutellarein, about 40 mg to about 500 mg scutellarein, about 40 mg to about 220 mg scutellarein, about 40 mg to about 130 mg scutellarein, about 40 mg to about 110 mg scutellarein, or about 40 mg to About 90 mg of scutellare About 0.25 g to about 3 g of scutellarin, about 0.3 g to about 3 g of scutellarin, about 0.3 g to about 1.5 g of scutellarin, about 0.3 g to about 0.9 g of scutellarin, about 0.3 g ~ About 0.8 g of scutellarin, or about 0.3 g to about 0.65 g of scutellarin. In some embodiments, the composition is used in the treatment of cancer, such as breast cancer and / or one or more gynecological cancers. In some embodiments, the cancer is breast cancer, such as advanced breast cancer, metastatic breast cancer, breast cancer resistant to treatment, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, and / or triple negative breast cancer.

  [0033] The inventor has also discovered a process for making a pharmaceutical composition using as a starting material a portion of Seitaanamikisou grown in the air. Such a process is particularly useful for making compositions comprising luteolin, apigenin, scutellarein, and scutellarin. Accordingly, in some embodiments, the inventor has described a process for making a pharmaceutical composition that includes: (a) water of a portion of the sorghum that grows in air above 40 ° C. Contacting for a period of at least about 10 minutes to form a mixture; (b) separating the portion of the sedum that grows in air to form a crude extract; (c) the crude extraction Separating high molecular weight compounds from the product to form a purified extract; (d) optionally evaporating some, substantially all, or all of the water from the purified extract, or purification thereof Additional water is added to the extracted extract; and (e) the purified extract is combined with at least one excipient other than pharmaceutically acceptable water to form the pharmaceutical composition. In some embodiments, the purified extract comprises apigenin, luteolin, scutellarein, and scutellarin. In some embodiments, the at least one pharmaceutically acceptable non-water excipient is selected from flavoring agents and sweeteners.

  [0034] The inventor has found that removing at least a part of the high molecular weight compound extracted from Seitakana myxo improves the clinical characteristics of the pharmaceutical composition. Since a large amount of soluble substances extracted from St. elegans is inactive, reducing the amount of soluble substances by removing molecules with a molecular weight above a predetermined cut-off value is The bulk of the pharmaceutical composition derived from the product will be greatly reduced. In addition, most of the soluble material extracted into water from Selexanthus is soluble fiber, which is not absorbed by the intestine and tends to promote stomach upset, abdominal distension, gas and diarrhea. Thus, removing at least a portion of its soluble fiber by reducing the burden of soluble material extracted from Selexanthus, while retaining a mixture of luteolin, apigenin, scutellarein, and scutellarin in the composition Resulting in improved anticancer drugs. Thus, in some embodiments, the invention provides a pharmaceutical composition comprising 1 part of a combination of luteolin, apigenin, scutellarein, and scutellarin and a high molecular weight compound having a molecular weight greater than a predetermined cut-off value of less than about 50 parts. Wherein the predetermined cutoff value is from 1,000 grams / mole to about 20,000 grams / mole. In some embodiments, the pharmaceutical composition comprises about 1 part luteolin, apigenin, scutellarein, and a combination of scutellarin, and less than about 40 parts, less than about 30 parts, less than about 20 parts, less than about 10 parts, about 5 parts. Less than, less than about 2 parts, less than about 1 part, less than about 0.5 part, from about 0.01 to about 40 parts, from about 0.01 to about 20 parts, or from about 0.01 to about 10 parts high molecular weight compound. Including. In some embodiments, the cutoff value for the high molecular weight compound is: 10,000 grams / mole; 5,000 grams / mole; 2,000 grams / mole; 1,000 grams / mole; or about 1, In the range of about 1,000 grams / mole to about 10,000 grams / mole; about 1,000 grams / mole to about 5,000 grams / mole or about 1,000 grams / mole to about 2,000 grams / mole. In some embodiments, the pharmaceutical composition includes at least one excipient other than water. In some embodiments, the at least one non-water excipient is a flavoring agent, sweetener, or both. The inventor has found that the pharmaceutical composition described herein is conveniently prepared as a dosage unit comprising the pharmaceutical composition described herein. In some embodiments, the dosage unit comprises a combination of at least about 18 mg luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the dosage unit comprises: about 0.25 g to about 4 g of luteolin, apigenin, scutellarein, and a combination of scutellarin; about 0.27 g to about 4 g of luteolin, apigenin, scutellarein, A combination of about 0.35 g to about 4 g of luteolin, apigenin, scutellarein, and scutellarin; a combination of about 0.35 g to about 2 g of luteolin, apigenin, scutellarein, and scutellarin; about 0.35 g to about 1 0.1 g of luteolin, apigenin, scutellarein, and scutellarin combination; from about 0.35 g to about 1 g of luteolin, apigenin, scutellarein, and scutellarin; from about 0.35 g to about 0.8 g of luteolin, apige Emissions, scutellarein, and combinations of Scutellarin; or about 0.7g~ about 2g of luteolin, apigenin, scutellarein, and combinations Scutellarin. In some embodiments, in addition to the combination of luteolin, apigenin, scutellarein, and scutellarin, the composition further comprises at least one non-water excipient. In some embodiments, the at least one non-water excipient is selected from a corrigent, sweetener, or both. In some embodiments, the present invention provides a method of treating cancer comprising administering to a cancer patient an effective amount of a pharmaceutical composition or dosage form described herein. In some embodiments, the cancer is one or more types of breast cancer and / or gynecological cancer, such as breast cancer or uterine cancer. In some embodiments, breast cancer, eg, advanced breast cancer, metastatic breast cancer, treatment resistant breast cancer, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, and / or triple negative breast cancer.

  [0035] The inventor has also discovered a process for making a pharmaceutical composition comprising: (a) water grown above 40 ° C. and a period of at least about 10 minutes in the air of Selexanthus To form a mixture; and (b) separating the portion of the cereals that grew in the air from the mixture to produce a crude extract; (c) producing a high molecular weight compound from the crude extract; Separating to form a purified extract; (d) optionally evaporating some, substantially all, or all of the water from the purified extract or adding to the purified extract And (e) combining the purified extract with a pharmaceutically acceptable excipient to form the pharmaceutical composition. 1 part luteolin, about 0.61 to about 2 parts apigenin, about 2.5 to about 9.4 parts scutellarein, and about 15 to about 70 parts scutellarin.

  [0036] Some embodiments also provide a process for making a purified extract of St. abyss, comprising: (a) heating a portion of St. abyss grown in air to above 40 ° C Contacting with water for a period of at least about 10 minutes to form a mixture; (b) separating the portion of the cereals that grew in the air from the mixture to produce a crude extract; and (c) The high molecular weight compound is separated from the crude extract to form a purified extract of Selexanthus.

  [0037] Some embodiments are further processes for making a pharmaceutical composition, wherein at least one pharmaceutically acceptable excipient other than water is one of the group consisting of luteolin, apigenin, scutellarein, and scutellarin A process comprising combining the above members to form a pharmaceutical composition thereof is provided. In some embodiments, the at least one non-water pharmaceutical excipient is selected from flavoring agents and sweeteners.

  [0038] Some embodiments provide a process for making a pharmaceutical dosage unit comprising: (a) at least a portion of water that is grown in air above 40 ° C. and at least water Forming a mixture upon contact for a period of about 10 minutes; (b) separating the portion of the cedar mushrooms grown in air to form a crude extract; and (c) the crude extraction Separating the high molecular weight compound from the product to form a purified extract; and (d) combining the purified extract with at least one excipient other than water to form the pharmaceutical dosage unit. To do. In some embodiments, the at least one non-water excipient is selected from flavoring agents and sweeteners.

  [0039] The term "about" followed by a specified value is intended to indicate a value within the experimental error of that specified value (generally within one standard deviation). Unless the experimental error is specifically determined, the term “about” followed by the specified value “x” may be taken to mean X ± 0.1X.

Process for the production of pharmaceutical compositions and unit dosages containing the extract of half-branched lotus
[0040] The pharmaceutical compositions and unit dosages described herein comprise a soluble substance (ie, a substance that is soluble in water) extracted from the portion of the half-branched, especially the bluefin beetle, grown in the air. Labiatae family herbaceous lame (Lamiaceae) -Ban Zhi Lian (BZL) is not exclusive, but mainly in the provinces of Sichuan, Jiangsu, Jiangxi, Fujian, Guangdong, Guangxi and Shaanxi It grows in the southeastern region of the Huang Po. The plant is harvested after the flower blooms (May-June) in late summer and early autumn. The part grown in the air is cut off from the roots. Only the portions grown in the air (leaves and stems) are used for the preparation of the compositions and dosage units described herein.

[0041] Table 1 shows the scientific names of the plants of the present invention from which the extracts of the present invention are obtained, listed by family, genus, species and traditional Chinese name.
Table 1

Pharmaceutical composition
[0042] Some embodiments described herein provide pharmaceutical compositions, particularly pharmaceutical compositions for the treatment of cancer. In particular, the present invention provides pharmaceutical compositions (“compositions”) for the treatment of gynecological cancers and breast cancer. In some preferred embodiments, the composition is for the treatment of breast cancer, particularly breast cancer (described in more detail below) that has been considered particularly difficult to treat for oncologists, including: Advanced breast cancer, metastatic breast cancer, breast cancer that is negative for one or more hormone receptors (eg, ER negative, PR negative, and / or HER2 negative breast cancer), and one or more previously used cancer therapies, such as Breast cancer that has been unsuccessfully treated with radiation therapy, proton therapy, and / or chemotherapy. The inventor has found that treatment of patients having one or more of these types of cancer with a soluble substance extracted from at least about 20 g of Selexanthus is well tolerated and effective in the treatment of these cancers. I found it.

  [0043] In some embodiments, the present invention provides a pharmaceutical composition comprising at least one excipient other than water and one or more members of the group consisting of luteolin, apigenin, scutellarein, and scutellarin. . Although each of luteolin, apigenin, scutellarein, and scutellarin have anti-cancer activity at the molecular level, all four combinations of these compounds are resistant to cancer, particularly breast cancer, and even previous treatments. It has been found to be particularly effective against breast cancer that has been found to be present. Thus, in some preferred embodiments, the composition comprises each of luteolin, apigenin, scutellarein, and scutellarin.

  [0044] Especially at the higher doses used in the methods of treating cancer described herein, the extract of Selexanthus may be unpleasant, which may even discourage patient compliance. I know it. Therefore, in order to make the composition more palatable and thereby increase patient comfort in the treatment and potentially increase patient compliance, flavoring agents such as flavorings or other flavoring agents, sweeteners It is desirable to use a fee or both. Accordingly, in some embodiments, the at least one excipient other than water is selected from a corrigent and a sweetener. As used herein, it is stated that a pharmaceutical composition contains, contains, or otherwise includes “excipients other than water”. If water is a suitable excipient for that particular form of dosage unit present therein, it may also contain water, but the composition may be any excipient apart from water. Means that it must contain. For example, some embodiments include a soluble material extracted from Selexanthus, a flavoring agent, and water. Other embodiments may further include a sweetener in addition to the corrigent, or may use a sweetener in place of the corrigent.

  [0045] The inventor has found that high doses of Selexanthus extract (eg, at least 20 g of soluble substance extracted from Selexanthus, or higher doses) may cause stomach upset, abdominal distension, gas and / or at least in some patients. They also found that it causes diarrhea. The inventor has found that high molecular weight compounds extracted from Selexanthus are inactive against breast and / or gynecological cancers and induce gastrointestinal distress, especially at doses of 20 g / day or more Decided to tend to. At the doses described herein, such stomach discomfort can result in poor patient compliance or even discontinuation of therapy for at least some patients. By removing at least a portion of the high molecular weight compound from the soluble material extracted from Sedum, such as by nanofiltration, the amount of soluble material that must be administered to the patient is reduced and extracted from Sedum It is believed that gastrointestinal discomfort associated with high concentrations of soluble substances will be reduced. Accordingly, the inventors herein provide teachings of compositions in which the high molecular weight compound is depleted and in some cases substantially free of it.

  [0046] The combination of luteolin, apigenin, scutellarein, and scutellarin, in another context, is not useful herein in the treatment of breast cancer and / or gynecological cancers, although not "high molecular weight compounds" and high molecular weight compounds. It may be referred to as an “effective soluble substance” as opposed to an “inactive soluble substance” that includes no compound. Thus, the mass of the soluble material is equal to the sum of the effective soluble materials (luteolin, apigenin, scutellarein, and scutellarin) and the inactive soluble material. The mass of the inert soluble substance is the sum of the masses of the high molecular weight compound and other inert compounds.

  [0047] As used herein, a "high molecular weight compound" is co-extracted with luteolin, apigenin, scutellarein, and scutellarin during the water extraction process of Selexanthus, with a predetermined cut-off. Refers to a compound having a molecular weight of or greater than. In some embodiments, the cutoff value may be anywhere from 1,000 g / mol to about 10,000 g / mol. In some embodiments, a cut-off value of 1,000 grams / mole will be sufficient to remove a high percentage of soluble fiber from a soluble extract of Solidago; however, a lower cut-off value is contemplated Lower cut-off values enable the achievement of higher concentrations of luteolin, apigenin, scutellarein, and scutellarin in pharmaceutical compositions and dosage units, and soluble substances that must be administered to patients to achieve a therapeutic effect In some cases, a lower cut-off value is preferred because it is believed to reduce the bulk. In some embodiments, the cutoff value is in the range of 750 to 20,000 g / mol, preferably 750 to 10,000 g / mol, and more particularly in the range of 750 to 5,000 g / mol. Individual cut-off values include: 750 g / mol; 1,000 g / mol; 2,000 g / mol; 5,000 g / mol; and 1,0000 g / mol.

  [0048] Accordingly, some embodiments of the compositions and dosage units provided herein are substantially free of high molecular weight compounds. As used herein, the term “substantially free” means that the composition or dosage unit has been treated (eg, filtered or decanted) to remove insoluble material (eg, stems, leaves and insoluble parts thereof). Less) than otherwise contained in the “crude extract”, which is a water extract of the portion grown in the air of St. abyss that has not been otherwise treated to remove high molecular weight compounds, It is meant to contain less high molecular weight compounds than some predetermined proportion. In some embodiments, the predetermined ratio is 1/10 (0.1), 1/20 (0.05), 1/50 (0.02), 1/100 (0.01), 1 / 200 (0.005), 1/500 (0.002) or 1/1000 (0.001). Individual values for “substantially free of high molecular weight compounds” can also be expressed in comparison to the total mass of soluble material extracted from St. abyss contained in the pharmaceutical composition. In some embodiments, the composition substantially free of high molecular weight compounds is less than about 10% by weight, less than about 5% by weight, about 1% by weight relative to the total mass of soluble material extracted from Sedum Less than, less than about 0.5% by weight, or less than about 0.1% by weight of high molecular weight compound. Furthermore, individual values for “substantially free of high molecular weight compounds” are expressed as mass ratios with respect to the amount of luteolin, apigenin, scutellarein, and scutellarin contained in the composition. In some embodiments, about 1% to about 99% of the soluble material extracted from Solidago is in the pharmaceutical composition is an effective soluble material, and from about 1.7% to about 3.2% of the effective soluble material. % Is luteolin, about 2% to about 3.4% is apigenin, about 7.9% to about 15.8% is scutellarein, and about 49% to the rest of the effective soluble substance is scutellarin. is there.

  [0049] In some cases, it may be sufficient to remove only a portion of the high molecular weight compound from the soluble material extracted from Sedum. Thus, in some embodiments of the compositions and dosage units provided herein, high molecular weight compounds are depleted. As used herein, the term “depleted” refers to a predetermined percentage that the composition or dosage unit is less than contained in the “crude extract” described in the previous paragraph. Is meant to contain less high molecular weight compounds. In some embodiments, the predetermined ratio is 9/10 (0.9), 8/10 (0.8), 7/10 (0.7), 6/10 (0.6), 1 / 2 (0.5), 1/3 (0.333) or 1/4 (0.25). Individual values for “high molecular weight compound depleted” can also be expressed in comparison to the total mass of soluble material extracted from St. abyss contained in the pharmaceutical composition. In some embodiments, the composition having a depleted amount of high molecular weight compound is less than about 90%, less than about 80%, less than about 70% by weight relative to the total mass of soluble material extracted from Sedum Less than about 60 wt% or less than about 50 wt% of high molecular weight compounds. In addition, individual values for “high molecular weight compound depleted” are expressed as mass ratios for the amount of apigenin, luteolin, scutellarein, and scutellarin contained in the composition. In some embodiments, about 1% to about 99% of the soluble material extracted from Solidago is in the pharmaceutical composition is an effective soluble material, and from about 1.7% to about 3.2% of the effective soluble material. % Is luteolin, about 2% to about 3.4% is apigenin, about 7.9% to about 15.8% is scutellarein, and about 49% to the rest of the effective soluble substance is scutellarin. is there.

  [0050] Active compounds (Luteolin, Apigenin, Scutellarein, and Scutellarin) extracted from Solidago are considered to be important for their joint activity in the water extract of the part of the Solidago found in the air It has been found that there is a tendency to be expressed in some specific ratio. In some embodiments, the combination of luteolin, apigenin, scutellarein, and scutellarin is about 1 part luteolin, about 0.61 to about 2 parts apigenin, about 2.5 to about 9.4 parts scutellarein, and about Contains 15 to about 70 parts scutellarin. In some embodiments, the combination of luteolin, apigenin, scutellarein, and scutellarin is about 1 part luteolin, about 0.75 to about 1.64 parts apigenin, about 3.1 to about 7.5 parts scutellarein, And about 20.4 to about 54.7 parts scutellarin. In some embodiments, the combination of luteolin, apigenin, scutellarein, and scutellarin is about 1 part luteolin, about 0.9 to about 1.3 parts apigenin, about 3.9 to about 6 parts scutellarein, and about Contains 37 to about 43 parts of scutellarin. In some embodiments, the composition comprises about 1 part luteolin, about 1.1 parts apigenin, about 4.8 parts scutellarein and about 34 parts scutellarin, comprising luteolin, apigenin, scutellarein, and scutellarine. Includes combinations.

The process of making a pharmaceutical composition
[0051] The pharmaceutical composition provided herein comprises extracting the active compound from, for example, water, from a portion (stems and / or leaves) of Sedum on the air. As described herein with respect to extraction, “water” refers to pure water (eg, water for injection, distilled water, double deionized water, filtered distilled water, etc.) and its extraction medium. Includes an aqueous solution composed of water and one or more trace amounts of solid or liquid solutes, as long as they are mostly water and the solute (s) do not substantially affect the extraction characteristics of the water. In some preferred embodiments, the process also includes removing a portion of the high molecular weight compound from the extract of Aspergillus as described in more detail above.

  [0052] Accordingly, in some embodiments, there is provided a process for making a pharmaceutical composition comprising: (a) at least a portion of water that is grown in the air of Selexiana and heated to above 40 ° C and at least Contacting for a period of about 10 minutes to form a mixture; (b) separating the portion of the cedar mellitus grown in air from the mixture to produce a crude extract; (c) the crude extract Separating the high molecular weight compound from the product to form a purified extract; and (e) combining the purified extract with at least one pharmaceutically acceptable non-water excipient to produce a pharmaceutical composition thereof. Form things. In some embodiments, the process also includes: (d) optionally evaporating some, substantially all, or all of the water from the purified extract, or the purified extraction Add additional water to the product. In some embodiments, the at least one pharmaceutically acceptable non-water excipient is selected from flavoring agents and sweeteners. In some embodiments, the pharmaceutical composition is further combined with a suitable package to form a suitable dosage unit.

  [0053] The portion (leaves and / or stem) grown in the air of Solidago is combined with water at a suitable temperature above room temperature, in particular from about 40 ° C, more preferably from about 50 ° C to about 80 ° C, In some cases, heating is performed at high pressure. The mixture should be cooked for a time that is not long enough for the active compound to be extracted into the aqueous phase of the mixture but unnecessarily wastes energy or causes decomposition of the active compound It is. A period of longer than about 10 minutes but shorter than about 2 days is suitable, but a period of 30 minutes to 6 hours is generally considered suitable. More detailed values are set forth in the examples herein.

  [0054] Once cooked, the portion of the sedum that grows in the air is separated from the aqueous phase by any suitable method. By rubbing the mixture through a strainer, a larger portion can be removed while a smaller portion can be removed by filtration. The filtration may be performed in stages, each stage including passing through one or more filters that are successively reduced in pore size.

[0055] High molecular weight compounds can be removed by any suitable method, such as nanofiltration or size exclusion chromatography.
[0056] In some cases, the volume of the solution can be reduced, for example, by evaporating and removing a portion of the water. The solution may be lyophilized, or dried in other situations to form a dry residue, which may be pulverized to form a powder. In any case, the resulting purified extract can then be combined with at least one excipient, particularly an excipient other than water, to form a pharmaceutical composition. In some embodiments, the non-water excipient is a corrigent or sweetener. In some preferred embodiments, the excipient other than water comprises a taste masking agent.

  [0057] Another embodiment provides a process for making a pharmaceutical composition comprising: (a) at least about 10 minutes with water heated above 40 ° C. of a portion of the sorghum that grows in air To form a mixture; and (b) separating the portion of the cedar mushrooms grown in air from the mixture to produce a crude extract; (c) high molecular weight from the crude extract. Are separated to form a purified extract; and (e) the purified extract is combined with a pharmaceutically acceptable excipient to form the pharmaceutical composition. Some embodiments also include: (d) optionally evaporating some, substantially all, or all of the water from the purified extract, or adding to the purified extract Add water. In some embodiments, the combination of luteolin, apigenin, scutellarein, and scutellarin is about 1 part luteolin, about 0.61 to about 2 parts apigenin, about 2.5 to about 9.4 parts scutellarein, and about Contains 15 to about 70 parts scutellarin. In some embodiments, the combination of luteolin, apigenin, scutellarein, and scutellarin is about 1 part luteolin, about 0.75 to about 1.64 parts apigenin, about 3.1 to about 7.5 parts scutellarein, And about 20.4 to about 54.7 parts scutellarin. In some embodiments, the combination of luteolin, apigenin, scutellarein, and scutellarin is about 1 part luteolin, about 0.9 to about 1.3 parts apigenin, about 3.9 to about 6 parts scutellarein, and about Contains 37 to about 43 parts of scutellarin. In some embodiments, the composition comprises about 1 part luteolin, about 1.1 parts apigenin, about 4.8 parts scutellarein and about 34 parts scutellarin, comprising luteolin, apigenin, scutellarein, and scutellarine. Includes combinations.

  [0058] In some embodiments, there is provided a process for making a composition comprising: (a) at least about 10 minutes with water heated above 40 ° C. of a portion of the sorghum that grows in air For a period of time to form a mixture; (b) separating the portion of the cereals that grew in the air from the mixture to produce a crude extract; and (c) producing a high extract from the crude extract. The molecular weight compound is separated to form a purified extract. The purified extract can be further processed to produce the dosage unit described herein. In some embodiments, the purified extract is depleted of high molecular weight compounds. In some embodiments, the purified extract is substantially free of high molecular weight compounds.

  [0059] As noted above, in certain circumstances, it may be desirable to mask the taste of the active compound contained in the extract of Selexanthus, particularly when the dosage is at least about 20 g / day. Accordingly, some aspects of the process of making a pharmaceutical composition include at least one pharmaceutically acceptable non-water excipient (eg, a corrigent and / or sweetener) from luteolin, apigenin, scutellarein, and scutellarin. Combining with one or more members of a group to form the pharmaceutical composition.

[0060] In some such embodiments, the at least one non-water pharmaceutical excipient is selected from flavoring agents and sweeteners.
Dosage Units
[0061] The inventor believes that the pharmaceutical compositions described herein are conveniently prepared in dosage units for convenient distribution, storage and administration. There is a difference between “dose” and “dosage unit” as described herein. As used herein, the term “dose” refers to the amount of the pharmaceutical composition that is administered in a single occurrence. A daily dose is the amount of the pharmaceutical composition administered daily. The dose is once a day (QD), twice a day (bid), three times a day (tid), four times a day (qid). ) And the like.

  [0062] As used herein, the term "dosage unit" is a single pre-manufactured form of the pharmaceutical composition, which is one or more doses of the pharmaceutical composition, or other dosage unit. In proportion to the dose of the pharmaceutical composition that can be combined to form a single dose. In some embodiments, the dosage unit consists of a single daily dose of the pharmaceutical composition. The dosage unit can be adapted to be administered as a single daily dose (QD), or two, three, four or more doses for administration at different times of the day (Bid, tid, or qid, respectively) or can be administered as a single dose. (This is particularly true for elixirs that can be divided into two or more doses per dosage unit and tablets that can be divided into two or more dosage units for administration at different times during the day. .) In some other embodiments, the dosage unit may comprise a proportion of a single dose (eg, half, one third, one quarter, one fifth). The dosage unit may be a solution for infusion of a specific volume, for example 20 mL to 1000 mL, for administration by a drip line or similar intravenous method, or even by a nasopharyngeal tube. Good.

[0063] Some preferred embodiments of the dosage unit include tablets, capsules, powders and solutions (elixirs).
[0064] Tablets include tablets to swallow, tablets to chew and swallow, and tablets adapted to dissolve and swallow on the tongue, with or without liquids that help swallow . Suitable excipients for tablets include binders, bulking agents, disintegrants, dispersants, glidants, anti-sticking and anti-caking agents, and flavoring and sweetening agents. It is.

  [0065] Capsules include capsules for swallowing as a whole and capsules adapted to be dissolved in liquid excipients such as water. Capsules also include capsules for opening and dissolving their contents in suitable excipients such as water. Suitable excipients for capsules include dispersants, bulking agents, flavoring agents and sweeteners.

  [0066] Powders include powders packaged in suitable containers for transport and storage, such as foil sachets, sealed vials, and the like. Suitable excipients for powders include dispersants, bulking agents, flavoring agents and sweeteners.

  [0067] Solutions include water-based solutions containing water, excipients other than water, and effective soluble substances (Luteolin, Apigenin, Scutellarein, and Scutellarin) extracted from Sedum. In a preferred embodiment, the solution is packaged in a suitable sealed container and is packaged with instructions for administering the solution to the patient. For intravenous administration, the water-based solution may or may not contain excipients other than water.

  [0068] The inventor believes that the compositions described herein should be administered to a patient at levels never previously considered and, importantly, the patient can tolerate it. I found it. Surprisingly, for example, the composition described herein can be administered to a patient at a higher dose, that is, at a dose greater than a soluble substance extracted from 10 or 12 grams of Selexanthus per day. I know. Surprisingly, this administration does not cause dose limiting toxicity at high doses, particularly from 20 grams per day to about 40 grams per day (especially at 20, 30, and 40 grams per day). Based on these clinical data, the inventor has shown that the maximum dose that can be tolerated is greater than 40 grams per day and even up to about 200 grams per day, perhaps up to about 100 grams per day. I guess. Accordingly, some embodiments described herein are pharmaceutical dosages comprising at least about 20 grams of an effective pharmaceutical ingredient comprising at least one member of the group consisting of apigenin, luteolin, scutellarein, and scutellarin. Provides a unit body. In some embodiments, the effective pharmaceutical ingredient comprises each of luteolin, apigenin, scutellarein, and scutellarin. In some preferred embodiments, the dosage unit is an oral dosage unit. In some preferred embodiments, the dosage unit further comprises at least one excipient other than water. In some preferred embodiments, the dosage unit comprises at least one excipient selected from a corrigent and sweetener. In some embodiments, the dosage unit contains at least about 20 grams of active pharmaceutical ingredient. In some embodiments, the dosage unit can be divided between two or more doses for administration on a single day.

  [0069] In some embodiments, the pharmaceutical dosage unit comprises at least about 20 grams of an effective pharmaceutical ingredient that includes a soluble substance extracted from St. abyss. The soluble material extracted from the Selexanthus includes one or more of apigenin, luteolin, scutellarein and scutellarin; preferably it includes all four of apigenin, luteolin, scutellarein and scutellarin. In a particularly preferred embodiment, the soluble material comprises each of apigenin, luteolin, scutellarein and scutellarin in approximately the following ratios: about 1 part luteolin, about 0.61 to about 2 parts apigenin, about 2.5 to about 9 .4 parts of scutellarein, and about 15 to about 70 parts of scutellarin; about 0.75 to about 1.64 parts of apigenin, about 3.1 to about 7.5 parts of scutellarein, and about 20.4 to about 54. .7 parts scutellarin from about 0.75 to about 1.64 parts apigenin, from about 3.1 to about 7.5 parts scutellarein, and from about 20.4 to about 54.7 parts scutellarin; About 1.3 parts apigenin, about 3.9 to about 6 parts scutellarein, and about 37 to about 43 parts scutellarin; or about 1 part luteolin, about 1.1 parts apigenin, about .8 parts Scutellarein and about 34 parts of Scutellarin. In some embodiments, the soluble material extracted from the Selexanthus is depleted or substantially free of high molecular weight compounds. In some embodiments, the dosage unit is an oral dosage unit (e.g., a tablet for swallowing, chewing or swallowing on the tongue, a capsule for swallowing, a capsule for swallowing, and opening the contents appropriately) Capsule for dissolving and swallowing in a suitable liquid excipient, capsule for dissolving in whole in a suitable excipient, powder for dissolving in a suitable excipient, such as a corrigent, sweetener and the like (Or may contain water). Thus, in some embodiments, the dosage unit further comprises at least one excipient other than water (eg, in addition to water). In some embodiments, the dosage unit comprises at least one excipient selected from a corrigent and sweetener. In some embodiments, the dosage unit comprises at least about 20 grams of the active pharmaceutical ingredient (e.g., 20-200 grams per dosage unit body, 20-100 grams per dosage unit body, or 20-20 grams per dosage unit body). 60 grams).

  [0070] In some embodiments, the pharmaceutical dosage unit comprises at least about 0.25 g of an effective pharmaceutical ingredient comprising a combination of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the pharmaceutical dosage unit comprises at least about 0.27 g luteolin, apigenin, scutellarein, and scutellarin or at least about 0.35 g luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the pharmaceutical dosage unit is about 0.35 g to 4 g, 0.35 g to 2 g, 0.35 g to 1.1 g, 0.35 g to 1 g, or 0.35 g to 0.8 g luteolin. , Apigenin, scutellarein, and scutellarin. In some embodiments, the pharmaceutical dosage unit is about 0.25 g, about 0.27 g, about 0.3 g, about 0.35 g, about 0.4 g, about 0.45 g, about 0.5 g, about 0. .6g, about 0.7g, about 0.8g, about 0.9g, about 1g, about 1.1g, about 1.2g, about 1.3g, about 1.4g, about 1.5g, about 1.6g About 1.7 g, about 1.8 g, about 1.9 g, about 2 g, about 2.1 g, about 2.2 g, about 2.3 g, about 2.4 g, about 2.5 g, about 2.6 g, about 2.7 g, about 2.8 g, about 2.9 g, about 3 g, about 3.1 g, about 3.2 g, about 3.3 g, about 3.4 g, about 3.5 g, about 3.6 g, about 3. 7 g, about 3.8 g, about 3.9 g, about 4 g of luteolin, apigenin, scutellarein, and scutellarin. Soluble material extracted from Sedum cane includes one or more of apigenin, luteolin, scutellarein and scutellarin; preferably it includes all four of apigenin, luteolin, scutellarein and scutellarin. In a particularly preferred embodiment, the soluble material comprises each of apigenin, luteolin, scutellarein and scutellarin in approximately the following ratios: about 1 part luteolin, about 0.61 to about 2 parts apigenin, about 2.5 to about 9 .4 parts of scutellarein, and about 15 to about 70 parts of scutellarin; about 0.75 to about 1.64 parts of apigenin, about 3.1 to about 7.5 parts of scutellarein, and about 20.4 to about 54. .7 parts scutellarin from about 0.75 to about 1.64 parts apigenin, from about 3.1 to about 7.5 parts scutellarein, and from about 20.4 to about 54.7 parts scutellarin; About 1.3 parts apigenin, about 3.9 to about 6 parts scutellarein, and about 37 to about 43 parts scutellarin; or about 1 part luteolin, about 1.1 parts apigenin, about .8 parts Scutellarein and about 34 parts of Scutellarin. In some embodiments, the soluble material extracted from the Selexanthus is depleted or substantially free of high molecular weight compounds. In some embodiments, the dosage unit is an oral dosage unit (e.g., a tablet for swallowing, chewing or swallowing on the tongue, a capsule for swallowing, a capsule for swallowing, and opening the contents appropriately) Capsule for dissolving and swallowing in a suitable liquid excipient, capsule for dissolving in whole in a suitable excipient, powder for dissolving in a suitable excipient, such as a corrigent, sweetener and the like (Or may contain water). Thus, in some embodiments, the dosage unit further comprises at least one excipient other than water (eg, in addition to water). In some embodiments, the dosage unit comprises at least one excipient selected from a corrigent and sweetener.

  [0071] The dosage unit described herein may be manufactured by a process according to the present invention. In some embodiments, there is provided a process for making a pharmaceutical dosage unit comprising: (a) at least about 10 minutes with water heated above 40 ° C. of a portion of the meal that has grown in air. To form a mixture; and (b) separating the portion of the cedar mushrooms grown in air from the mixture to produce a crude extract; (c) high molecular weight from the crude extract. Are separated to form a purified extract; and (d) the purified extract is combined with at least one excipient other than water to form the pharmaceutical dosage unit. In some embodiments, the at least one non-water excipient is selected from flavoring agents and sweeteners. Such a dosage unit contains an amount of purified extract suitable for treating cancer, particularly breast cancer. In some embodiments, the dosage unit comprises a combination of at least 0.25 g, at least 0.27 g, at least 0.3 g, at least 0.35 g, or 0.35 g to 4 g of luteolin, apigenin, scutellarein, and scutellarin. . In some embodiments, the dosage unit further comprises a package, such as a foil sachet, bottle, small sachet, blister pack, or other sealed package. Thus, in some embodiments, the process of making the dosage unit includes packaging the dosage unit in a package.

  [0072] Illustrative amounts of each dosage unit according to the present invention, or of the effective soluble substances (Luteolin, Apigenin, Scutellarein, and Scutellarin) in each dose are shown in Table 2 below.

How to use
[0073] The pharmaceutical compositions and dosage units described herein can be used to treat cancer, particularly breast cancer and gynecological cancer. The inventor has conducted a clinical trial in humans of the composition according to the present invention and administration of soluble substances extracted from 20 grams per day, 30 grams per day or 40 grams per day is a well-tolerated substance. It has been found effective against breast cancer, particularly breast cancer with advanced breast cancer that has previously received at least one round of cancer therapy, at least one round of chemotherapy. Treatment-resistant breast cancer is particularly difficult to treat and the inventor has provided a method for treating cancer in humans. In particular, the inventors have provided a method of treating breast cancer in humans in addition to providing a method of treating one or more cancer subtypes, including metastatic breast cancer. Other cancers that can be treated include cancers that do not express estrogen receptor (ER negative breast cancer), cancers that do not express progesterone (PR negative breast cancer), cancers that do not express human epidermal growth hormone receptor 2 (HER2 negative breast cancer) ) Is included. In this regard, it is noted that these breast cancer categories are not mutually exclusive. For example, the breast cancer can be ER negative and PR negative (so-called double negative breast cancer) or ER negative, PR negative and HER2 negative (triple negative breast cancer). Triple negative breast cancer may be advanced and / or metastatic. Metastatic breast cancer may, and often will, have been found to be resistant to one or more previous therapeutic approaches. Thus, as used herein, listing one feature of breast cancer (eg ER negative) is not intended to exclude other features (eg PR negative) unless explicitly stated otherwise.

  [0074] Accordingly, some embodiments of the invention comprise an effective amount of at least one excipient other than water and at least one member of the group consisting of luteolin, apigenin, scutellarein, and scutellarin for cancer patients. Provided is a method of treating cancer comprising administering a pharmaceutical composition. In some embodiments, the composition comprises each of apigenin, luteolin, scutellarein, and scutellarin, wherein the at least one non-water excipient is selected from flavoring agents and sweeteners. In some embodiments, the composition is substantially free of high molecular weight compounds. In some embodiments, the cancer is breast cancer or gynecological cancer. In some embodiments, the cancer is breast cancer that is at least one of the following: advanced breast cancer, metastatic breast cancer, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, ER negative and PR negative breast cancer, ER negative PR-negative and HER2-negative breast cancer, or breast cancer that has not responded to at least one previous series of cancer treatments.

  [0075] Some embodiments of the present invention further provide a method of treating cancer, eg, breast cancer or gynecological cancer, wherein the patient suffering from the cancer is at least about 0.25 g, at least about 0.27 g, Methods are provided by administering a pharmaceutical composition comprising 0.3g, at least about 0.35g, or about 0.35g to about 4g of a combination of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the method applies to the patient about 0.25 g, about 0.27 g, about 0.3 g, about 0.35 g, about 0.4 g, about 0.45 g, about 0.5 g, about 0.6 g. About 0.7 g, about 0.8 g, about 0.9 g, about 1 g, about 1.1 g, about 1.2 g, about 1.3 g, about 1.4 g, about 1.5 g, about 1.6 g, about 1.7 g, about 1.8 g, about 1.9 g, about 2 g, about 2.1 g, about 2.2 g, about 2.3 g, about 2.4 g, about 2.5 g, about 2.6 g, about 2. 7 g, about 2.8 g, about 2.9 g, about 3 g, about 3.1 g, about 3.2 g, about 3.3 g, about 3.4 g, about 3.5 g, about 3.6 g, about 3.7 g, Administering a daily dose of a combination of about 3.8 g, about 3.9 g, about 4 g of luteolin, apigenin, scutellarein, and scutellarin. In some embodiments, the cancer is breast cancer or gynecological cancer. In some embodiments, the cancer is breast cancer that is at least one of the following: advanced breast cancer, metastatic breast cancer, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, ER negative and PR negative breast cancer, ER negative PR negative and HER2 negative breast cancer, or breast cancer that has not responded to at least one previous course of cancer treatment.

  [0076] As noted above, the inventor conducted clinical trials, and doses in excess of soluble material extracted from 20 grams per day of Selexanthus are well tolerated, especially in cancer patients in difficult-to-treat groups I found it effective. In addition, the inventor has found that the active compound in the soluble substance of the extract of Selexanthus is one or more (preferably all four) of apigenin, luteolin, scutellarein, and scutellarin. Accordingly, some embodiments comprise administering to a patient a pharmaceutical dosage unit comprising an effective pharmaceutical ingredient comprising at least 20 grams of apigenin, luteolin, scutellarein, and at least one member of the group consisting of scutellarin. A method of treating cancer is provided. In some embodiments, the effective pharmaceutical ingredients include each of apigenin, luteolin, scutellarein, and scutellarin. In some embodiments, the dosage unit is an oral dosage unit. In some embodiments, the dosage unit further comprises at least one excipient other than water. In some embodiments, the dosage unit comprises at least one excipient selected from a corrigent and sweetener. In some embodiments, the method includes at least about 0.25 g, about 0.27 g, about 0.3 g, about 0.35 g, about 0.4 g, about 0.45 g, about 0.5 g, about 0.1. 6 g, about 0.7 g, about 0.8 g, about 0.9 g, about 1 g, about 1.1 g, about 1.2 g, about 1.3 g, about 1.4 g, about 1.5 g, about 1.6 g, About 1.7 g, about 1.8 g, about 1.9 g, about 2 g, about 2.1 g, about 2.2 g, about 2.3 g, about 2.4 g, about 2.5 g, about 2.6 g, about 2 0.7 g, about 2.8 g, about 2.9 g, about 3 g, about 3.1 g, about 3.2 g, about 3.3 g, about 3.4 g, about 3.5 g, about 3.6 g, about 3.7 g About 3.8 g, about 3.9 g, about 4 g of a soluble substance extracted from Solidago, including a combination of luteolin, apigenin, scutellarein, and scutellarin Comprising administering a. In some embodiments, the cancer is breast cancer that is at least one of the following: advanced breast cancer, metastatic breast cancer, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, ER negative and PR negative breast cancer, ER negative PR negative and HER2 negative breast cancer, or breast cancer that has not responded to at least one previous course of cancer treatment.

  [0077] Some embodiments described herein include an effective pharmaceutical ingredient comprising at least one member of the group consisting of apigenin, luteolin, scutellarein, and scutellarin at least 20 grams per day. A method of treating cancer comprising administering. In some embodiments, the active pharmaceutical ingredient is administered at a dose of 1 to 4 per day. In some embodiments, the cancer is breast cancer or gynecological cancer. In some embodiments, the cancer is breast cancer that is at least one of the following: advanced breast cancer, metastatic breast cancer, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, ER negative and PR negative breast cancer, ER negative PR negative and HER2 negative breast cancer, or breast cancer that has not responded to at least one previous course of cancer treatment. In some embodiments, the method includes at least about 0.25 g, about 0.27 g, about 0.3 g, about 0.35 g, about 0.4 g, about 0.45 g, about 0.5 g, about 0.1. 6 g, about 0.7 g, about 0.8 g, about 0.9 g, about 1 g, about 1.1 g, about 1.2 g, about 1.3 g, about 1.4 g, about 1.5 g, about 1.6 g, About 1.7 g, about 1.8 g, about 1.9 g, about 2 g, about 2.1 g, about 2.2 g, about 2.3 g, about 2.4 g, about 2.5 g, about 2.6 g, about 2 0.7 g, about 2.8 g, about 2.9 g, about 3 g, about 3.1 g, about 3.2 g, about 3.3 g, about 3.4 g, about 3.5 g, about 3.6 g, about 3.7 g About 3.8 g, about 3.9 g, about 4 g of a soluble substance extracted from Solidago, including a combination of luteolin, apigenin, scutellarein, and scutellarin Comprising administering a.

  [0078] Some embodiments described herein comprise administering to a patient a pharmaceutical dosage unit comprising an effective pharmaceutical ingredient comprising a soluble substance extracted from at least 20 grams of Selexus. A method of treating cancer is provided. In some embodiments, the dosage unit is an oral dosage unit. In some embodiments, the dosage unit further comprises at least one excipient other than water. In some embodiments, the dosage unit comprises at least one excipient selected from a corrigent and sweetener. In some embodiments, the dosage unit comprises at least about 20 grams of soluble material extracted from Selexanthus.

  [0079] Some embodiments further provide a method of treating cancer comprising administering to a patient an effective pharmaceutical ingredient comprising a soluble substance extracted from at least 15 grams of Selexanthus per day. In some embodiments, the active pharmaceutical ingredient is administered at a dose of 1 to 4 per day. In some embodiments, the cancer is breast cancer or gynecological cancer. In some embodiments, the cancer is breast cancer that is at least one of the following: advanced breast cancer, metastatic breast cancer, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, ER negative and PR negative breast cancer, ER negative PR negative and HER2 negative breast cancer, or breast cancer that has not responded to at least one previous course of cancer treatment.

  [0080] The inventor has found that in some embodiments, the individual doses used to treat the patient are important for successful clinical outcome. Thus, in some embodiments, the patient must be administered at least 20 g / day of soluble material extracted from Sedum. In some embodiments, the dosage unit contains at least about 20 grams of active pharmaceutical ingredient. In some embodiments, the dosage unit is about 20 grams to about 200 grams, about 20 grams to about 100 grams, about 20 grams to about 60 grams, about 20 grams, about 30 grams, about 40 grams, about 50 grams. About 60 grams, about 70 grams, about 80 grams, about 90 grams or about 100 grams of active pharmaceutical ingredient. The preferred mode of administration is oral administration, preferably wherein the soluble material extracted from the sedum is combined with at least one excipient other than water, such as a corrigent, sweetener or both.

In vitro activity of an extract of Selexanthus
[0081] Table 3A shows the extent of inhibition of the activity of several in vitro solid breast cancer tumor cell lines by the extracts of this invention.

  [0082] Table 3B shows the degree of inhibition of the activity of several in vitro solid cancer tumor cell lines by the extracts of this invention.

  [0083] It is an aspect of the present invention to isolate and characterize the active compound in an extract from Solidago ("BZL"). The extract loses activity when reconstituted after drying and when the extract is separated by physical and chemical means.

  [0084] As used herein, the terms "treat", "treating" and "treatment" refer to ameliorating one or more symptoms of a disease state. Successful treatment may be judged by achieving stable disease, partial or complete remission, or partial or complete delay in disease progression. One suitable endpoint for successful treatment is prolonging life expectancy.

  [0085] As used herein, "administer", "administering" or "administration" refers to the extract (s) of the invention, or the extract (single or Delivery of a pharmaceutical composition comprising a plurality of) to a patient in a manner suitable for the treatment of the particular cancer being addressed.

[0086] "Patient" refers to a mammal having a tumor, particularly a human, and more particularly a female human suffering from one or more gynecological cancers or breast cancers.
[0087] The terms "effective amount" and "therapeutically effective amount" as used herein, (1) reduce tumor size in a patient population; (2) inhibit tumor metastasis (ie, to some extent) (3) Suppress tumor growth to some extent (ie slow down, preferably stop) to some extent; and / or; (4) Relieve one or more symptoms associated with cancer to some extent (or (5) Stabilize tumor growth; (6) Prolong time to disease progression; (7) Synonymous with the amount of composition or dosage unit that has the effect of improving overall survival. Point to it.

  [0088] A "pharmaceutical composition" as used herein is described herein with a mixture of one or more compounds or other chemical components, such as physiologically acceptable carriers and excipients. Refers to a combination. The purpose of the pharmacological composition is to facilitate administration of the extract (s) of this invention to a patient.

[0089] The term "pharmaceutically acceptable" as used herein means that the agent or excipient is generally considered acceptable for use in a pharmaceutical composition.
[0090] As used herein, a "physiologically acceptable carrier" is a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered composition. Point to. Typical pharmaceutically acceptable carriers include solid and liquid diluents. Water, ethanol, propylene glycol, and glycerol are illustrative pharmaceutically acceptable liquid diluents; among these, water is preferred in some embodiments.

  [0091] As used herein, "excipient" refers to a pharmaceutically inert substance added to a pharmaceutical composition to further facilitate administration of the pharmaceutical composition of this invention. Examples of excipients include, but are not limited to calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols. Excipients and groups of active pharmaceutical ingredients are considered mutually exclusive in the pharmaceutical arts. In some preferred embodiments, the excipient is a flavoring agent, sweetener, or both.

  [0092] The term "excipient other than water" means that the excipient is or includes any excipient other than water, such as a corrigent or sweetener. Thus, the term “excipients other than water” will include excipients containing water and sweeteners or water and flavoring agents, but will exclude excipients containing only water. A pharmaceutical composition comprising an excipient other than water and an active pharmaceutical ingredient includes, for example, the pharmaceutically active ingredient, water, and any other excipient, such as a corrigent and / or sweetener. May contain.

  [0093] As used herein, the terms "comprising", "comprises", "comprise" and their grammatical variations are inclusive or open-ended. ) And does not exclude additional, non-enumerated elements or method steps. The terms “include”, “includes”, “contain”, “contains”, “containing” and their grammatical variants are similarly inclusive.

[0094] As used herein, the phrase "consisting of" excludes any element, step, or ingredient not specified in the following part of the sentence.
[0095] As used herein, the phrase “consisting essentially of” refers to a substance or process that specifies the scope of the subsequent portion of the sentence and the basic and novel features of the claimed invention. Limited to substances or processes that do not substantially affect (one or more).

  [0096] As used herein, "BZL" is synonymous with "Seitakanamikisou". The term “BZL101” refers to a specific extract of BZL that has demonstrated activity against cancer cells. In particular, it is intended to be the part of Seita Kanamikisou grown in the air.

  [0097] Herbs from which the extracts of this invention were obtained were purchased from Shen Non Herbs, Berkeley, California. Their identity was confirmed by reference to traditional pharmaceutical literature.

Preparative Example 1-Isolation of active compound from half-branched lotus (BZL)
[0098] The dried half lotus was extracted with 8: 2 MeOH—H 2 O for 6 and 12 hours. The combined extracts were filtered, concentrated in vacuo, and partitioned sequentially with hexane and ethyl acetate (equal volume, repeated once). Combine the ethyl acetate portions and concentrate in vacuo and run on a Sephadex lipophilic LH-20 medium (approx. 160 g, 1800 × 25 mm ID column) with homogeneous solvent 9: 0.5: 0.5 MeOH-acetone-H 2 O or Chromatography under gravity flow with 100% MeOH. Fractions (40 ml) were collected and combined based on analytical HPLC (Table 3) and / or RF-TLC (1: 1 10 mM ammonium acetate-MeCN) analysis.

  [0099] Scutellarein (1), isoscutellarein (2), luteolin (3), and apigenin (4) were eluted from the Sephadex LH-20 column in the partially overlapped fraction. Individual compounds were purified using preparative HPLC method A (Table 3).

  [0100] The flavanones Carthamidin (5) and Isocartamidine (6) were eluted together from the Sephadex LH-20 column in a fraction different from the flavones described above. Cartamidine (5) and isocartamidine (6) were purified using preparative HPLC method B (Table 3).

  [0101] Isoscutellarein was identified based on LC / MS and 1D and 2D NMR analysis. All other compounds (1, 3-6) were identified based on LC / MS and NMR comparisons with a commercially available reference standard or a synthetically proven standard. NMR spectra were recorded using a Varian Mercury Plus 400 MHz. The HPLC and UV spectra were recorded using an Agilent Technologies 1200 Series HPLC system equipped with a DAD detector and using a Phenomenex Luna C18 (150 × 2.1 mm, 3 μm) column. The molecular mass was determined in negative mode using Applied Agilent Technologies 6210 TOF LC / MS. A summary of the characteristics of the equipment used is shown in Table 1-1.

  [0102] Table 1-1: HPLC method. The columns listed below were used in the isolation of compounds 1-6. The same solvent gradient was used for the chromatography for all HPLC runs and only the flow rates were different as specified. Gradient: Solvent A: 0.1% TFA. Solvent B: MeCN; linear gradient without upfront hold in 30 minutes from 10% B to 60% B.

[0103] The NMR data used to identify compounds 1-6 are described below.
[0104] Scutellarein (1): CAS # 529-53-3; LC / MS [MH] -m / z 285.0425. Formula 1 shows the important HMBC correlation of compound (1). NMR data for Compound 1 is shown in Table 1-2.

  [0105] Isoscutellarein (2): CAS # 41440-05-5; LC / MS [M-H] -m / z 285. Formula (2) shows an important HMBC correlation of compound (2). NMR data relating to Compound 2 are shown in Table 1-3.

  [0106] Luteolin (3): CAS # 491-70-3; LC / MS [M-H] -m / z 285.0403. Equation 3 shows the important HMBC correlation of compound (3). The NMR data for compound 3 are shown in Table 1-4.

  [0107] Apigenin (4) CAS # 520-36-5; LC / MS [M-H] -m / z 269.04479. Formula (4) shows the structure of apigenin (4).

  [0108] Cartamidine (5): CAS # 479-54-9; LC / MS [M-H] -m / z 287. Formula (5) shows the important HMBC correlation of compound (5). NMR data for Compound 5 are shown in Table 1-5.

  [0109] Isocartamidine (6): CAS # 2569-76-8; LC / MS [M-H] -m / z 287. Formula (6) shows the important HMBC correlation of compound (6). NMR data for Compound 6 are shown in Table 1-6.

Preparative Example 2-Preparation of BZL101 for human in vivo experiments
[0110] BZL101 is an aqueous extract of a portion of the family Lamiaceae that grew in the air. Herbaceous Seita Kanamikisou (Chinese Pinyin translation-Ban Zhi Lian (BZL)) grows mainly in the southeastern region of Huang Po in Sichuan, Jiangsu, Jiangxi, Fujian, Guangdong, Guangxi and Shaanxi provinces. The plant is harvested after the flowers bloom in late summer and early autumn. The part grown in the air (leaves and stems) is cut off from the root and used as starting material (BZL). The portion of the herb grown in the air is dried in the sun and wrapped as whole grass. To ensure purity, the herb is identified by botanical, morphological and chemical characterization to confirm authenticity.

[0111] A single dose of BZL101 is made by the following procedure and is referred to as BZL101 (Bionovo, Inc., Emeriville, CA).
・ 180g raw grass is ground to a fine powder (25 mesh)
• Mix the powder with 1800 ml of distilled water to form a slurry • Boil the slurry at 70-72 ° C. for 60 minutes • Decant the extract and filter through a 22 μm filter • Weight of the supernatant after extraction The volume of the solution is 1750 ml. The extract is concentrated in 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 product is 12 grams. • Pack it in a sterile, vacuum-sealed container. • Tests for bacteria, yeast and heavy metals are performed by an accredited laboratory.

  [0112] For higher doses (eg, 20, 30, and 40 grams per day), the raw material herb (the portion of the sedum moth that grows in the air and the amount of water is weighed proportionally, resulting in Proportional dry weight of soluble material.

Example 1: Characterization of the active substance from Steller's root
[0113] BZL101 induces cell death in breast cancer cells, but does not induce cell death in non-transformed breast epithelial cells. This selective cytotoxicity is based on the strong induction of reactive oxygen species (ROS) in tumor cells by BZL101. As a result, cancer cells treated with BZL101 develop extensive oxidative DNA damage and die with necrotic death. Data from expression profiling of cells treated with BZL101 strongly supports the death pathway, including oxidative stress, DNA damage and activation of genes that promote death. In breast cancer cells, oxidative damage induced by BZL101 leads to poly (ADP-ribose) polymerase (PARP) overactivation and subsequent reduced levels of NAD persistence and ATP depletion, both of which are cancerous. Not observed in cells that are not. Since inhibition of PARP results in suppression of necrosis and activation of the apoptotic death program, PARP overactivation has become a means of a necrotic death program induced by BZL101. BZL101 treatment results in selective inhibition of glycolysis in tumor cells, which is evident from a decrease in enzyme activity within the glycolysis pathway and inhibition of lactate production. Since tumor cells often rely on glycolysis for energy production, the observed inhibition of glycolysis is likely to be an important factor in the selective energy collapse and necrotic death in breast cancer cells . The expected selectivity of BZL101 for cancer cells is based on metabolic differences between highly glycolytic tumor cells and normal cells.

[0114] Several types of experiments were performed using individual compounds isolated from BZL101.
[0115] A total of seven purified compounds from BZL101 were tested for several biological activities present throughout the aqueous BZL101 extract: induction of ROS, DNA damage and cell death. The following parameters were examined:
1. Induction of loss of mitochondrial membrane potential difference (MTP). All compounds tested induced loss of MTP.

  2. Reactive oxygen species (ROS) induction. Cell permeability indicators of induced ROS, such as dihydroethidium (specific for superoxide) (FIG. 2), CM-H2DCFDA (most types of ROS) (FIG. 1) and MitoSOX (superoxide derived from mitochondria) ( The fluorescence from FIG. 3) was examined using a fluorescence plate reader and FACS.

  3. Induction with either specific indicators for mitochondrial ROS and / or specific inhibitors of ROS production by known sources such as mitochondria, ubiquinone oxidoreductase NQO (mitochondrial complex I) and NADPH oxidases The compounds were also tested for potential cellular sources of ROS.

4). For induction of DNA damage, compounds were tested using a test known as the comet assay that allows detection of DNA damage in individual cells. (Fig. 4)
5. Induction of death in cells treated with the compound was investigated using the propidium iodide test for cell permeability and subsequent analysis in FACS.

  6). The mode of cell death (ie apoptosis vs. necrosis) was investigated using several criteria: conversion of cells to be positive for binding to Annexin V; DNA fragmentation (characterized by death in apoptosis) and Reduced cellular ATP levels (commonly observed during necrotic death) (FIG. 5). 1. Induction of loss of mitochondrial membrane potential difference (MTP). All compounds tested induced loss of MTP.

  2. Reactive oxygen species (ROS) induction. Cell permeability indicators of induced ROS, such as dihydroethidium (specific for superoxide) (FIG. 2), CM-H2DCFDA (most types of ROS) (FIG. 1) and MitoSOX (superoxide derived from mitochondria) ( The fluorescence from FIG. 3) was examined using a fluorescence plate reader and FACS.

  3. Induction with either specific indicators for mitochondrial ROS and / or specific inhibitors of ROS production by known sources such as mitochondria, ubiquinone oxidoreductase NQO (mitochondrial complex I) and NADPH oxidases The compounds were also tested for potential cellular sources of ROS.

4). For induction of DNA damage, compounds were tested using a test known as the comet assay that allows detection of DNA damage in individual cells. (Fig. 4)
5. Induction of death in cells treated with the compound was examined for cell permeability using the propidium iodide test followed by FACS analysis.

  6). The mode of cell death (ie apoptosis vs. necrosis) was investigated using several criteria: conversion of cells to be positive for binding to Annexin V; DNA fragmentation (characterized by death in apoptosis) and Reduced cellular ATP levels (commonly observed during necrotic death) (FIG. 5).

  The data shown in FIGS. 1-5 is summarized in Table 1-7 below.

[0117] Two breast cancer cell lines, MDA MB231 and SKBr3, were used in the experiments summarized in Tables 1-7 with similar results.
result
[0118] All of the compounds tested induced a loss of mitochondrial membrane potential and ranged from 25 to 90% loss of MTP compared to mock-treated cells (not shown). .

  [0119] Most of the compounds have cytotoxic activity; however, each has a different degree of cytotoxicity (Tables 1-7). The compound has different effects on the induction of reactive oxygen species (ROS), DNA damage and cellular energy status (FIGS. 1-5). Thus, BZL101 extract contains several compounds that may differ in the mode of cell death induction.

[0120] Analysis of the mechanism of induction of death by different compounds reveals at least two different modes of cytotoxicity:
[0121] All compounds tested induce cellular ROS within minutes after treatment, which is an oxidant-sensitive fluorescent probe 5- (and -6) -chloromethyl-2 ', 7'-dichlorodihydrofluorescein diacetate It was determined by loading the cells using acetyl ester (CM-H ₂ DCFDA or DCFDA). DCFDA is non-fluorescent in reduced form and easily penetrates the membrane. Cellular esterases cleave their acetate groups. The thiol-reactive chloromethyl group then binds to cellular thiols and traps the dye inside the cell where oxidation converts it to a fluorescent form. CM-H ₂ DCFDA is oxidized by cellular hydrogen peroxide, hydroxyl radicals, and various free radical products downstream from hydrogen peroxide. It is relatively insensitive to superoxide oxidation. However, since hydrogen peroxide is produced by disproportionation of superoxide, CM-H ₂ DCFDA can be used as an indirect indicator of superoxide production. As can be seen in FIG. 1, CM-H ₂ DCFDA is oxidized intracellularly by all BZL101 compounds tested, but the levels of total ROS induced are different.

  [0122] All compounds tested also induced superoxide, which was determined by staining the cells with dihydroethidium, a cell-penetrating indicator that was selectively oxidized by superoxide. Cytosolic dihydroethidium exhibits blue fluorescence; however, once this probe is oxidized by superoxide to ethidium, it intercalates inside the cell's DNA, staining its nucleus in a bright fluorescent red color, It is easily detected by flow cytometry methods (Figure 2).

[0123] Two flavonoids, apigenin and luteolin, induce the production of superoxide whose origin is identified as mitochondria. MitoSOX, a specific detection agent for mitochondrial superoxide, was converted to its fluorescent form by apigenin and luteolin, but not by other compounds. In addition, inhibitors of mitochondrial respiration (sodium azide, NaN ₃ ) and inhibitors of mitochondrial complex I (dicumarol, not shown) prevented the production of superoxide by apigenin and luteolin (FIG. 3).

  [0124] Apigenin and luteolin differ from other compounds in that they do not induce DNA damage (Figure 4). However, both are cytotoxic and characterized as apoptotic based on binding to annexin V, DNA fragmentation, and a slight but consistent increase in ATP levels observed during the first hours of treatment. Induces significant cell death (Figure 5). All these features are apoptotic death characteristics.

  [0125] The five compounds identified as either tetrahydroxyflavones (scutellarein, isoscutellarein, cartamidine and isocartamidine) or pentahydroxyflavone (no name) have a different mechanism for cell death. Induce. They induce ROS but are from outside mitochondria (the source of which has not yet been determined). These compounds also induce DNA damage, the extent of which seems to correlate with the level of induction of ROS. Types of ROS induced by scutellarein and similar compounds, such as singlet oxygen, may be particularly effective in inducing DNA damage. Since superoxide is not membrane permeable and cannot induce direct oxidative DNA damage, it makes sense to assume that most ROS induced by these compounds is not superoxide. However, superoxide can be rapidly converted to peroxide in the cell, which can directly damage DNA.

  [0126] Like all BZL extracts, the five compounds mentioned above induce a reduction in cellular ATP levels. Loss of ATP with no or low staining for Annexin V is more consistent with necrotic cell death.

Figure legend:
FIG. _CM-H 2 DCFDA was determined by staining with the induction of ROS in SKBr3 cells. The indicated compounds were added to the cells in 20 μg / ml growth medium followed by 10 μM CM-H ₂ DCFDA. NaN3, an inhibitor of mitochondrial respiration, was added at 10 mM. After a 30 minute incubation, cells were washed in PBS and analyzed on a FACScan for fluorescence. The names of the compounds are abbreviated in this and other figures as follows: A-apigenin; C-cartamidine; L-luteolin; S-scutellarein; IC-isocartamidine; A species with a molecular weight of -320 (possibly pentahydroxyflavone).

  FIG. Induction of superoxide in SKBr3 cells as determined by staining with dihydroethidium. The indicated compounds were added to the cells followed by 5 μM dihydroethidium. After a 20 minute incubation, cells were washed in PBS and analyzed on a FACScan for fluorescence.

FIG. Cells were stained with MitoSOX, an indicator of mitochondrial superoxide. Processing was performed as described in the legend for FIG.
[0130] FIG. Induction of DNA damage in SKBr3 cells by compounds isolated from BZL101 was analyzed using the comet assay. Cells were treated with the indicated compounds at 20 μg / ml for 15 minutes and analyzed using the comet assay kit from Trevigen according to the manufacturer's instructions. Briefly, cells were collected, washed and resuspended in PBS. The cells were mixed with low melting point agarose thawed at 37 ° C. and pipetted onto comet slides. Agarose was allowed to solidify at 4 ° C. for 30-40 minutes and soaked in cold lysis solution (Trevigen, Inc.) at 4 ° C. for 30 minutes. The slide was soaked in freshly prepared alkaline solution (300 mM NaCl and 1 mM EDTA) for 20 minutes and electrophoresed at 300 mA in the same alkaline buffer for 30-40 minutes. Slides were rinsed in water and then fixed in 70% ethanol for 5 minutes. After air drying, nuclei were stained with Sybr green (Trevigen, Inc.) and observed under a fluorescence microscope. The percentage of cells with comets was quantified by an observer who did not know the identity of the slide.

  [0131] FIG. SKBr3 cells were plated on 96 well plates and treated with the indicated compounds for 4 hours. Cells were lysed in situ and ATP content was analyzed on a 96-well plate-based luminometer using an ATP Bioluminescence Assay Kit HSII from Roche.

Conclusion:
[0132] The BZL101 extract contains a compound having cytotoxic activity. Although these compounds have different effects on mitochondrial and cellular DNA, all have cytotoxic activity against human cancer cells. Two of the identified compounds, apigenin and luteolin, induce mitochondrial superoxide and apoptotic death, which is carried out by mitochondrial or endogenous pathways.

  [0133] The other five compounds, particularly scutellarein and isoscutellarein, induce ROS, followed by DNA damage and cell death, which has the characteristics of programmed necrosis.

Example 2-Separation and synergistic activity of an active substance extracted from Steller's root
[0134] As demonstrated in Example 2, several compounds extracted from Sedum myrtle have been shown to induce reactive oxygen species (ROS) production, DNA damage and cell death. In order to better understand the combined activity of the isolated species, several flavanones and flavones isolated from half-branches were tested individually and in combination. The tested flavanones and flavones are shown in FIG. These compounds 1-8 were tested as described in Example 2 for induction of ROS, DNA damage and cell death. The results of these tests are shown in Tables 10 and 11 below.

  [0135] As can be seen in Tables 10 and 11, the combination of luteolin and isoscutellarein is much more effective than the same concentration of luteolin alone in generating reactive oxygen species (ROS) and inducing cell death. Similarly, the combination of luteolin and isoscutellarein is much more effective than the same concentration of isoscutellarein alone in generating reactive oxygen species (ROS) and inducing cell death. In this sense, the combination of isoscutellarein and luteolin is believed to have a synergistic effect on ROS production and cell death induction.

  It is also clear from Tables 10 and 11 that the combination of luteolin and apigenin is much more effective than the same concentration of luteolin alone in generating reactive oxygen species (ROS) and inducing cell death. Similarly, the combination of luteolin and apigenin is much more effective than the same concentration of apigenin alone in generating reactive oxygen species (ROS) and inducing cell death. In this sense, the combination of apigenin and luteolin is believed to have a synergistic effect on ROS production and induction of cell death.

  [0137] As can be seen in Tables 10 and 11, the combination of apigenin and isoscutellarein is much more effective than the same concentration of apigenin alone in generating reactive oxygen species (ROS) and inducing cell death. Similarly, the combination of apigenin and isoscutellarein is much more effective than the same concentration of isoscutellarein alone in generating reactive oxygen species (ROS) and inducing cell death. In this sense, the combination of isoscutellarein and apigenin is believed to have a synergistic effect on ROS production and cell death induction.

[0138] The results shown in Tables 10 and 11 were confirmed by conducting experiments in two different breast cancer cell lines.
Example 4-In vivo efficacy of an active substance derived from BZL101 in humans
[0139] In order to demonstrate the safety and clinical activity of BZL101 orally, a combination of active compounds isolated from Sedum is tested in human patients with advanced breast cancer.

  [0140] Eligible patients have histologically confirmed metastatic breast cancer and measurable disease. Patients are not given any other chemotherapy, hormonal therapy or herbal medicine during the trial. Patients take 350 ml of drug per day (equal to 0.00001-1 gram of each of the 1, 2, 3, 4, 5 or all members of the group consisting of apigenin, luteolin, scutellarein, and scutellarin) Are given until disease progression, toxicity or personal preference has discontinued them. The primary endpoints are safety, toxicity and tumor response.

  [0141] Patients are registered and given medication. Record the average age and the average number of previous treatments. Track and record any hematological and Grade III or IV non-hematological adverse events (AEs). Carefully observe and record Grade I and II adverse events such as nausea, diarrhea, headache, flatulence, vomiting, constipation, and fatigue, if any. Patients who can be assessed for response are evaluated, and patients with disease stability (SD) greater than 90 days and patients with SD greater than 180 days are carefully observed and recorded. Patients with minor objective tumor regression should also be carefully observed and recorded.

[0142] Patients are registered at one or more appropriate research centers and sign informed consent approved by the local institutional review board. The following excludes patients from the study: large liver involvement (greater than 50% of liver parenchyma), lymphatic vessels that have not been stabilized by therapy for more than 3 months Pulmonary concomitant, central nervous system concomitant or spinal cord compression, history of multiple or severe food or drug allergies, and creatinine greater than 2.0 g / dl, total bilirubin greater than 1.7 g / dl Organ or bone marrow dysfunction defined by a white blood cell count of less than 2,500 cells / μL and a platelet count of less than 75,000 mm ³ . ("Dl" = deciliter (s))
[0143] Safety monitoring is performed on an ongoing basis, and the patient meets a physician for examination at baseline and every Y weeks. Adverse events are graded using the second edition of the Common Toxicity Criteria, assigned categories by organ system, remote, potentially related, and potentially related It is encoded as probable or definitely related. Baseline tumor assessments will be performed within 14 days of study drug initiation and every 3 months. The response is evaluated using the RECIST criteria. Study medication was given at each visit, and compliance and doses taken at this visit were reviewed. The test drug is provided as a liquid in a sealed and labeled aluminum packet containing a complete daily dose administered in divided doses twice a day. The study drug is administered daily until it is determined that it has faced tumor progression or dose limiting toxicity, or until the subject decides to voluntarily discontinue, in which case the reason for discontinuation is obtained.

result
[0144] The results of the above test are recorded and evaluated based on meeting the test endpoint.

Example 4-Concentration of active substance in soluble substance extracted from Sedum
[0145] BZL101 is prepared as described herein. The active compounds, luteolin, apigenin, scutellarein, and scutellarin are identified and quantified with respect to 1 mg of BZL101. The respective masses of luteolin, apigenin, scutellarein, and scutellarin in 1 mg of the soluble substance in BZL101 are shown in Table 4-1.

  [0146] As can be seen in Table 4-1, in this illustrative and non-limiting example, 1 mg of soluble material extracted from Sedum is about 0.44 μg ± 0.05 μg luteolin, 0.49 μg ± Contains 0.04 μg apigenin, 2.1 μg ± 0.2 μg scutellarein and 15 μg ± 2 μg scutellarin. Thus, each mg of dry soluble material extracted from Selexanthus contains about 18 μg ± 5 μg of the combination of luteolin, apigenin, scutellarein, and scutellarin in a ratio of about 1: 1.1: 4.8: 34.

Example 5-Selexanthus Extract in the Treatment of Refractive Metastatic Breast Cancer for Treatment
[0147] Treatment of patients with metastatic breast cancer was performed using an extract of Selexanamussou ("BZL101"). Extract BZL101 was prepared essentially as described above and was given to patients who received one or more courses of metastatic breast cancer. BZL101 was given either once a day (qd) or twice a day (bid) as described below. The 20 gram, 30 gram, and 40 gram doses have been found to be well tolerated even though they are much higher than previously reported in the literature on half-branches. In addition, some patients have shown efficacy as discussed below.

  [0148] BZL101 is an extract of the half-branched lotus, which shows a novel mechanism of action. Normal cells rely on the citrate cycle (> 85%) and glycolysis (<7%) for energy production. Cancer cells rely on glycolysis (> 85%) for energy production. BZL101 inhibits energy production by inhibiting glycolysis. BZL101 causes DNA damage and cancer cell death. BZL101 does not cause cell death in normal cells.

  [0149] The following principles have been proposed for the selective cytotoxic activity of BZL101 in cancer cells: Tumor cells rely on glycolysis for energy production. This is associated with increased endogenous reactive oxygen species (ROS) levels. Normal cells rely on oxidative phosphorylation for their energy needs. BZL101 treatment further increases the level of ROS in tumor cells, leading to polyADP ribose polymerase (PARP) overactivation and severe oxidative DNA damage. In normal cells, BZL101 treatment results in only moderate increases in ROS levels and moderate DNA damage without PARP activation.

  [0150] Activation of PARP depletes storage of NAD + / NADH (substrate for the synthesis of poly ADP-ribose) and ATP. Glycolysis uses cytosolic NAD + as a substrate to generate ATP, which is inhibited by NAD + deficiency. (Oxidative phosphorylation uses mitochondrial NAD + to generate ATP and is usually unaffected by PARP activation). The depletion of NAD + and ATP due to BARP activation induced by BZL101 results in inhibition of glycolysis, as well as reduced ATP levels and cell death. Breast Cancer Res Treat. 2007 Sep; 105 (1): 17-28. Epub 2006 Nov 17. PMID: 17111207; Cancer Biol Ther. 2008 Jan 7; 7 (4) [electronic version prior to print] PMID: 18305410.

  [0151] The main features of the study outlined in Example 3 and the latest phase 1B study (Example 5) are compared in Table 5-1 below.

The main features of the BZL101 Phase 1B cancer trial are summarized as follows:
Planning for BZL101 Phase 1B
[0152] First:
Determine the maximum tolerated dose of BZL101 Provide preliminary data about the safety and efficacy of BZL101 Secondly:
-Tumor response as defined by RECIST (response criteria in solid tumors)-Overall and progression free survival-Duration of response-Participant reported change in QOL (EORTC QLQ-C30)
[0153] Key eligibility criteria • Must have histologically confirmed breast cancer • Must have measurable stage IV disease • No more than 3 preceding for metastatic disease Chemotherapy (the first one was not limited in number and was corrected to a maximum of 3 in the middle of the trial)
[0154] A summary of the incrementally increasing capacities is shown in Table 5-2 below:

  [0155] Baseline characteristics for patients participating in the study are as shown in Table 5-3 below:

  [0156] The demographic breakdown of study participants is summarized in the following Table 5-4:

  [0157] The number and type of adverse events experienced by study participants are shown in Table 5-5 below:

[0158] Definition of Phase 1B dose limiting toxicity:
(A) Grade 3, 4, or 5 toxicity based on NCI CTCAE V 3.0, possibly related, likely related, or reliably related to study drug administration (b) Grade 2 gastrointestinal toxicity that may be related, likely, or certainly related to study drug administration and lasts for longer than 3 weeks (c) May be related to study drug administration Baseline laboratory or medical condition that is likely to be related, likely related, or definitely related, worsening to grade 3 or higher
[0159] The dose limiting toxicity (DLT) experienced by study participants is described in the following Tables 5-6:

[0160] Summary of Phase 1B adverse events:
a) BZL 101 is well tolerated. The most common relevant adverse events are: diarrhea (48%), nausea (40%), vomiting (26%) and fatigue (22%).

  b) There were 12 serious adverse events in the study and only one was considered related to study drug administration: hospitalization for Grade 3 rib pain due to vomiting at a dose of 40 g / day.

c) There were 3 patients with DLT:
(I) Increase in grade 4 AST,
(Ii) Grade 3 diarrhea and grade 3 fatigue in the same patient, and (iii) Grade 3 rib pain due to vomiting.

  [0161] Compliance with study drug administration is set forth in Tables 5-7 below:

[0162] Preliminary efficacy of Phase 1B:
a) 21 out of 27 people continued the study for more than 28 days b) 8/21 (38%) remained stable for over 90 days c) 4/21 (19%) over 180 days D) 18 out of 27 were evaluable by RECIST (at least one measurable lesion, follow-up scan completed or incomplete)
e) 6/18 (33%) was stable for over 90 days f) 3/18 (17%) was stable for over 180 days
[0163] These results are summarized in the following Tables 5-8:

Phase 2 evaluation items Major achievements
[0164] Obtain a preliminary estimate of efficacy based on the extent of tumor response using the RECIST criteria
[0165] Adverse events assessed by self-report, physical examination and laboratory results at each visit
[0166] Secondary outcomes a. Tumor response: degree of clinical benefit, complete response, partial response, disease progression b. Response duration and survival: overall response, complete and partial response duration, overall survival, and progression free survival c. Changes in quality of life with EORTC QLQ-C30 Summary
[0167] The reached MFD was 40 g / day. The second phase will enroll 80 patients (40 HR + and 40 HR-) and will proceed at 20 g / day.

[0168] Half-branch extracts inhibit breast cancer cell growth in vitro.
[0169] BZL101 treatment results in inhibition of glycolysis and inhibition of lactate production that is evident from a decrease in enzyme activity within the glycolysis pathway.

[0170] BZL101 causes selective cell death in cancer cells and not healthy cells.
[0171] Oral administration of BZL101 is well tolerated. The most common adverse events are: diarrhea (48%), nausea (40%), vomiting (26%) and fatigue (22%).

  [0172] There were 3 patients with DLT: Grade 4 AST elevation, Grade 3 diarrhea and Grade 3 fatigue in the same patient, and Grade 3 rib pain due to vomiting.

[0173] One SAE was thought to be due to BZL101; hospitalization for grade 3 rib pain due to vomiting at 40 g / day.
[0174] On average, compliance with study drug administration was 90% of the prescribed ingested dose.

  [0175] In this severely pre-treated population, 7/18 (39%) was stable for over 90 days, and 4/18 (22%) was stable for over 180 days. there were.

  [0176] Of the 27 women enrolled, 18 were progression, 3 were patient choices, 2 were AEs, 2 were SAEs, and 1 was non-compliant with test procedures Canceled.

  [0177] From the foregoing, it is noted that the extract of St. abyss administered in a dry weight of 20 grams, 30 grams, or 40 grams of metastatic breast cancer, particularly metastatic breast cancer that has been shown to be resistant to treatment. It can be understood that it is effective for treatment and well tolerated.

  [0178] From the above, a daily dose of 15 grams to 60 grams of dry weight of an extract of Selexanthus is ER negative breast cancer, PR negative breast cancer, Her2 / neu negative breast cancer and / or triple negative breast cancer ( (Including those that have metastasized to other tissues). These doses are also considered useful for the treatment of other ER negative, PR negative, Her2 / neu negative and triple negative cancers. The dry weight doses of 20, 30 and 40 grams per day are the aforementioned cancers, especially ER negative breast cancer, PR negative breast cancer, Her2 / neu negative breast cancer and / or triple negative breast cancer (those breast cancers that have metastasized to other tissues). Are considered particularly useful in the treatment of

  [0179] Additional clinical trials of BZL101 can be performed according to the methodology described in Example 4. A patient diagnosed with cancer is treated as described in Example 4 with appropriate modifications depending on the disease being treated, and 20 grams dry weight, 30 grams dry weight or 40 grams dry weight of BZL101. Treat and evaluate by weight (or any other amount greater than 15 grams dry weight, eg, about 15-60 grams dry weight). Typical cancers treated include adrenocortical cancer, anal cancer, aplastic anemia, bile duct cancer, bladder cancer, bone cancer, bone metastasis, adult CNS brain tumor, pediatric CNS brain tumor, breast cancer, Castleman's disease, cervix Cancer, childhood non-Hodgkin lymphoma, colon and rectal cancer, endometrial cancer, esophageal cancer, Ewing family tumor, eye cancer, gallbladder cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, gastrointestinal trophoblastic disease, Hodgkin Disease, Kaposi's sarcoma, kidney cancer, laryngeal and hypopharyngeal cancer, acute lymphocytic leukemia, acute myeloid leukemia, childhood leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, liver cancer, lung cancer, lung carcinoid tumor Non-Hodgkin lymphoma, male breast cancer, malignant mesothelioma, multiple myeloma, myelodysplastic syndrome, nasal cavity and sinus cancer, nasopharyngeal cancer, neuroblastoma, oral cavity and Oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, pituitary tumor, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma (adult soft tissue cancer), melanoma skin cancer, Non-melanoma skin cancer, gastric cancer, testicular cancer, thymic cancer, thyroid cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom macroglobulinemia, cancer derived from viruses and cancer related to viruses included.

General conclusion
[0180] While preferred embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Those skilled in the art will now be able to conceive numerous variations, modifications and substitutions without departing from the invention. It should be understood that various alternatives may be used in the practice of the invention for the aspects of the invention described herein. The following claims are intended to define the scope of the invention, and the methods and configurations within these claims and their equivalents are intended to be included therein.

Claims (20)

A dosage unit comprising at least about 20 g of soluble material extracted from Scutellaria barbata D. Don.   The dosage unit according to claim 1, further comprising at least one excipient.   3. A dosage unit according to claim 2, comprising at least one taste masking agent, at least one sweetener, or both.   2. A dosage unit according to claim 1 in a form suitable for oral administration.   The dosage unit according to claim 4, further comprising water.   2. A dosage unit according to claim 1 comprising from about 20 g to about 200 g of soluble material extracted from Sedum.   7. A dosage unit according to claim 6, comprising from about 20 g to about 40 g of soluble material extracted from Sedum.   The dosage unit of claim 1, wherein the soluble material extracted from St. abyssal comprises at least about 0.27 g of a combination of luteolin, apigenin, scutellarein, and scutellarin.   2. The dosage unit of claim 1, wherein the soluble material extracted from Solidago is about 0.35 g to about 4 g of a combination of luteolin, apigenin, scutellarein, and scutellarin. The dosage unit of claim 1, wherein the soluble material extracted from Solidago is about 0.35 g to about 0.8 g of a combination of luteolin, apigenin, scutellarein, and scutellarin.   A method of treating cancer comprising administering to a cancer patient at least about 20 g of soluble material extracted from Selexanthus per day.   12. The method of claim 11, wherein the cancer is selected from breast cancer and one or more gynecological cancers.   13. The method of claim 12, wherein the cancer is breast cancer.   14. The method of claim 13, wherein the breast cancer is advanced breast cancer, metastatic breast cancer, treatment resistant breast cancer, ER negative breast cancer, PR negative breast cancer, HER2 negative breast cancer, and / or triple negative breast cancer.   12. The method of claim 11, comprising administering to the patient a soluble material extracted from about 20 g per day to about 200 g per day.   16. The method of claim 15, comprising administering to the patient a soluble material extracted from about 20 g per day to about 40 g per day.   12. The method of claim 11, wherein the patient achieves stable disease, partial remission, or complete remission.   12. The method of claim 11, wherein the soluble material extracted from Solidago is at least about 0.27 g of a combination of luteolin, apigenin, scutellarein, and scutellarin.   12. The method of claim 11, wherein the soluble material extracted from Solidago is about 0.35 g to about 4 g of a combination of luteolin, apigenin, scutellarein, and scutellarin.   12. The method of claim 11, wherein the soluble material extracted from the Selexanthus includes about 0.35 g to about 0.8 g of a combination of luteolin, apigenin, scutellarein, and scutellarin.

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