JP6945587B2 - Methods of Inhibiting TIE2 Kinase Useful for Cancer Treatment - Google Patents

Description

(Explanation of the text file submitted electronically)
The entire contents of the text file submitted electronically with the present specification are incorporated herein by reference in their entirety. Computer-readable format copy of the sequence listing (file name: DECP_066_00WO_SeqList_ST25.txt, recording date: November 7, 2013, file size 6 kilobytes).

The present invention relates to methods of inhibiting TIE2 kinase useful in the treatment of tumor growth, tumor invasion, tumor invasion, tumor dissemination, tumor metastasis and tumor immunotolerance. Specifically, the present invention comprises the composition of formula I described herein as a potent inhibitor of TIE2 for treating breast cancer growth, invasion, intravascular invasion, dissemination, metastasis and immune tolerance. Regarding the method to use.

Intimal endothelial cell kinase 2 (TIE2) is largely restricted to expression in a subset of vascular endothelial cells and bone marrow-derived TIE2-expressing monocytes (TEM). TIE2 is a receptor for angiopoietin 1 (ANG1), angiopoietin 2 (ANG2) and angiopoietin 4 (ANG4), and this signaling system is angiogenesis (new blood vessel sprouting from existing blood vessels) and blood vessels. It plays an important role in both formation (new angiogenesis from the beginning). TEM is a subset of circulating monocytes and tissue macrophages with angiogenesis-promoting and angiogenesis-promoting activity in tumor models (De Palma MD et al, Cancer Cell 2005; 8: 211-226) (non-). Patent Document 1). Inhibition of TIE2 reduces the ability of TEMs associated with blood vessels (Mazzieri R, Cancer Cell 2011; 19: 512-526) (Non-Patent Document 2) and significantly reduces the angiogenesis-promoting activity of this macrophage subset. (De Palma M, Clin Cancer Res 2011; 17 (16): 5226-5232) (Non-Patent Document 3).

Cytotoxic chemotherapy, radiation therapy and anti-angiogenic therapy damage the tumor-related vasculature and thus result in a hypoxic tumor environment. This hypoxic tumor environment results in a rebound of tumor angiogenesis by activating the angiogenesis switch from the vascular endothelial growth factor (VEGF) / VEGFR2 pathway to the ANG / TIE2 pathway in vascular endothelial cells. Recruitment of angioplastic TEMs from the bone marrow to these hypoxic tumor sites facilitates this angiogenesis by association of endothelial cells with TEMs within the tumor microenvironment. Therefore, TEM and TIE2-expressing endothelial cells are thought to play an important role in tumor revascularization after these treatments, leading to proliferative progression of the remaining tumor cells (De Palma M, et al. Trends Immunol). 2007; 28: 519-524) (Non-Patent Document 4).

TIE2 is also a mediator of osteoclast differentiation, and TIE2 inhibition resulted in reduced osteolytic bone erosion and reduced tumor growth in the breast cancer model of 4T1 mice (
1557475887355_0
, Et al. Int J Oncol 2003; 22: 391-397) (Non-Patent Document 5). In addition to the physiological expression of TIE2 in the endothelium, monocytes / macrophages and osteoclasts of the tumor microenvironment, TIE2 has also been demonstrated to be present in breast cancer cells. Tumor cell expression of TIE2 was associated with an increased risk of metastatic disease and an independent predictor of prognosis in multivariate analysis (Min Y, et al. Cancer Res 2010; 70: 2918-2828) (Non-Patent Documents). 6).

Surprisingly, a subset of TIE2-expressing tissue macrophages are localized within a specific vascular structure known as the tumor microenvironment (TMEM) of metastases. Recent observations have linked TIE2-expressing macrophages within the TMEM structure to be essential for extravasation of breast cancer cells into the vascular circulation and subsequent dissemination to distal metastatic sites (Condeelis J, Pollard JW. Cell 2006; 124: 263-6; Ginter PS, et al. Cancer Res 2012; 72 (24 Suppl): Abstract # P6-02-04) (Non-Patent Documents 7 and 8). Therefore, inhibition of macrophages within the TIE2 and TMEM structures may result in a reduction in new metastases.

Recently, TIE2-expressing tissue macrophages (TEMs) have been described to play a role in breast cancer immune tolerance. Breast cancer-derived TEMs can suppress tumor-specific immune responses. Specifically, the inhibitory function of TEM is driven in the same manner as by TIE2 and VEGF kinase activity. TEMs isolated from breast cancer tissue can function as antigen-presenting cells that induce only weak proliferation of T cells. Inhibition of TIE2 and VEGFR kinase activities induced TEM to change its phenotype into cells with the characteristics of bone marrow dendritic cells with strong antigen presentation. The immunosuppressive activity of TEM is also associated with high CD86 surface expression and broad association of regulatory T cells in breast tumors. TIE2 and VEGFR kinase activities were required to maintain high CD86 surface expression levels and convert T cells to immunosuppressive regulatory cells (Ibberson M, et al. Clin Cancer Res 2013; 19: 3439-3449). ) (Non-Patent Document 9).

The polyoma middle T antigen (PyMT) syngeneic mouse breast cancer model utilizes a mouse breast cancer virus (MMTV) promoter, a mammary gland-specific promoter to express PyMT in mouse mammary gland tissue. In this model, PyMT breast cancer cells are implanted in a mouse breast fat pad and these cancers metastasize, resulting in the death of this mouse. The PyMT model, not limited to the xenograft model, utilizes fully immune-responsive mice. Metastases in this model are known to be regulated by TIE2-expressing macrophages within the TMEM vascular structure. Therefore, there is a need for new treatments for TIE2-related diseases.

De Palma MD et al, Cancer Cell 2005; 8: 211-226 Mazzieri R, Cancer Cell 2011; 19: 512-526 De Palma M, Clin Cancer Res 2011; 17 (16): 5226-5232 De Palma M, et al. Trends Immunol 2007; 28: 519-524 Dales JP, et al. Int J Oncol 2003; 22: 391-397 Min Y, et al. Cancer Res 2010; 70: 2918-2828 Condeelis J, Pollard JW. Cell 2006; 124: 263-6 Ginter PS, et al. Cancer Res 2012; 72 (24 Suppl): Abstract # P6-02-04 Ibberson M, et al. Clin Cancer Res 2013; 19: 3439-3449

式Ｉ
式中、
ｎは、０〜７の整数であり、
Ｘは、薬学的に許容され得る塩の塩基性ラジカルであり、
ただし、ｎが０である場合、前記式Ｉの組成は、親遊離塩基であるという条件である。一部の実施形態では、ＨＸは存在せず、これにより、前記式Ｉの構造は、前記親遊離塩基である。 The methods of the present invention find utility in inhibiting TIE2 kinase. As a result of this inhibition, the present invention is useful in treating or preventing tumor growth, invasion, intravascular invasion, dissemination, metastasis and tumor immune tolerance. In particular, the present invention relates to methods of using the composition of Formula I described below as a potent inhibitor of TIE2 for treating breast cancer growth, invasion, intravascular invasion, dissemination, metastasis and immune tolerance. ..

Formula I
During the ceremony
n is an integer from 0 to 7 and
X is a pharmaceutically acceptable salt basic radical,
However, when n is 0, the composition of the formula I is a condition that it is a parent free base. In some embodiments, HX is absent, whereby the structure of formula I is the parent free base.

The composition of Formula I also finds usefulness in other cancers. In this case, TIE2 expression in either the tumor cells or the tumor microenvironment is the growth of the primary tumor, the invasion of the primary tumor, the intravascular invasion of the tumor into the bloodstream, the dissemination of the tumor cells, the tumor into the distal tissue. Tumor progression is caused by a mechanism that mediates the metastasis of the tumor or the immune tolerance of the tumor. Thus, TIE2 kinase inhibition by the composition of Formula I described above is the growth of the primary tumor, the invasion of the primary tumor, the intravascular invasion of the tumor into the bloodstream, the dissemination of tumor cells, the metastasis of the tumor to distal tissues or the tumor. Finding usefulness in the treatment of cancer by inhibiting processes involving immune tolerance.

TIE2 kinase is glioma (Liu et al, Oncotarget (2010) 1: 700-709; Brunkhorst et al, Cancer Research (2010) 70: 7283-7293), melanoma (Helfrich et al, Clin Cancer Res (2009)). 1384-1392), ovarian cancer (Karan et al, J. Clinical Oncology (2012) 30: 362-370), colon cancer (Ahmad et al, Cancer (2001) 92: 1138-1143; Hashizume et al, Cancer Research (Cancer Research). 2010) 70: 2213-2223), hepatocellular carcinoma (Matsubara et al, Hepatology (2013) 57: 1416-1425; Mitsuhashi et al, Hepatology (2003) 37: 1105-1113; Tanaka et al, J. Clin. 1999) 103: 341-345) and blood cancer (Muller et al, Leukemia Research (2002) 26: 163-168; Hou et al, Leukemia Research (2008) 32: 904-912) can cause cancer progression. It is shown.

FIG. 1 shows the growth inhibition of primary PyMT tumors using the composition of Formula II, paclitaxel or a combination thereof. FIG. 2 shows inhibition of PyMT tumor macrophage accumulation using the composition of Formula II, paclitaxel or a combination thereof. FIG. 3 shows inhibition of TIE2-expressing cells in PyMT tumors using the composition of Formula II, paclitaxel or a combination thereof. FIG. 4 shows inhibition of lung metastasis in a PyMT breast cancer model using the composition of Formula II, paclitaxel or a combination thereof. FIG. 5 shows inhibition of lung metastasis in a PyMT breast cancer model comparing the activity of paclitaxel and the combination of paclitaxel with the composition of Formula II. FIG. 6 shows the inhibition of lung metastases in a PyMT breast cancer model using etibrine as a single agent or in combination with the composition of Formula II. FIG. 7 shows enzymatic and in vivo activity using etibrine as a single agent or in combination with the composition of Formula II.

Definitions The term "pharmaceutically acceptable basic radical of a salt" in the composition of Formula I above is not limited and is not limited to water-soluble and water-insoluble salts, such as substituted or unsubstituted benzene sulfonates, acetates. , Amsonate (4,4-diaminostylben-2,2-disulfonate), benzoate, bicarbonate, bicarbonate, heavy tartrate, borate, bromide, butyrate, calcium salt , Calcium edetate, camcillate, carbonate, chloride, citrate, clavularate, dihydrochloride, edetate, edicylate, estolate, esylate, fiunarate, Gluceptate, Gluconate, Glutamate, Glycolyl alsanylate, Hexafluorophosphate, Hexylresorcinate, Hydrabamine salt, Hydrobromide, Hydrochloride, Hydroxynaphthoate, Iodine, Isothion Acids, lactates, lactobionates, laurates, magnesium salts, malate, maleates, mandelates, mesylates, methyl bromides, methyl nitrates, methyl sulfates, mutinates, Napsylate, nitrate, N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate, pamoate (1,1-methene-bis-2-) Hydroxy-3-naphthoate, einbonate), pantothenate, phosphate / diphosphate, picphosphate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, Includes stearate, basic acetate, succinate, sulfate, sulfosalicylate, suramate, tannate, tartrate, theocrate, tosylate, triethiodide and valerate. Specific examples of the basic radicals include para-toluenesulfonate, triflate and methanesulfonate.

The term "salt" means a pharmaceutically acceptable salt.

The term "pharmaceutically acceptable salt" also means a salt of the composition of the invention having an acidic functional group, such as a carboxylic acid functional group and a base.

The term "treating" with respect to a subject means ameliorating at least one symptom of the subject's disorder. Treatment can be to cure, ameliorate, or at least partially recover the disorder.

As used in this disclosure, the terms "administer," "administering," or "administration" refer to the composition of said compound or a pharmaceutically acceptable salt or composition. It means both direct administration to the subject or administration of a prodrug derivative or analog or composition of the composition of the compound or a pharmaceutically acceptable salt to the subject. The prodrug derivative or analog may form an equivalent amount of active compound in the body of the subject.

When used for medical applications, an "effective amount" is an amount that is effective in providing measurable treatment, prevention, or reduction of the incidence of a disease of interest.

The present invention relates to methods of treating (blocking) or preventing tumor growth, invasion, intravascular invasion, dissemination, metastasis and tumor tolerance. The method comprises administering an effective amount of the composition of Formula I described herein to a patient in need of a treatment, reduction or prophylactic effect of these symptoms in a dosing regimen that regulates TIE2 inhibition. include.

The amount of composition described herein required to achieve a therapeutic effect can be empirically determined based on conventional methods for a particular purpose. Generally, the therapeutic agent is pharmacologically intended to administer a therapeutic agent (eg, a composition of formula I or II (and / or a further agent) described herein) for therapeutic purposes. Provided in effective doses. A "pharmacologically effective amount", a "pharmacologically effective dose", a "therapeutically effective amount" or a "effective amount" is an amount sufficient to produce the desired physiological effect, or In particular, it means an amount that can achieve the desired result for treating the disorder or disease. As used herein, an effective amount, for example, delays the progression of the disorder or signs of the disease, alters the course of the signs of the disorder or disease (eg, slows the progression of the signs of the disease), said. It shall contain an amount sufficient to reduce or eliminate one or more signs or symptoms of the disorder or disease and reverse the signs of the disorder or disease. For example, administration of a therapeutic agent to a patient with cancer is not only when the underlying disease is eradicated or ameliorated, but also when the patient has a reduction in the severity or duration of the symptoms associated with the disease. For example, reporting a decrease in tumor mass, a decrease in circulating tumor cells, and an increase in progression-free survival also provides therapeutic benefits. Therapeutic benefits also include stopping or delaying the progression of the underlying disease or disorder, whether or not improvement has been achieved.

式ＩＩ In one embodiment of the invention, the composition of formula I is 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazol-5-yl) -3- (2-yl) of formula II. Fluoro-4- (2- (methylcarbamoyl) pyridin-4-yloxy) phenyl) urea para-toluenesulfonate, which is a potent inhibitor of TIE2, a receptor tyrosine kinase for angiopoietin ligands. Is.

Formula II

The composition of Formula I is that TIE2 expression in either the tumor cells or the tumor microenvironment is the growth of the primary tumor, the invasion of the primary tumor, the intravascular invasion of the tumor into the bloodstream, the dissemination of the tumor cells, the distal. We find usefulness in cancer that causes tumor progression by a mechanism that mediates tumor metastasis to tissues or tumor immune tolerance. Thus, inhibition of TIE2 kinase by the composition of Formula I described in the growth of primary tumors, invasion of primary tumors, intravascular invasion of tumors into the bloodstream, dissemination of tumor cells, metastasis of tumors to distal tissues or Find usefulness in the treatment of cancer by inhibiting processes including tumor immunotolerance.

The composition of the above formula I at therapeutic concentrations inhibits cells in the tumor microenvironment known to cause tumor growth, invasion, intravascular invasion, dissemination, metastasis or tumor-induced immune tolerance. Such cell types in the tumor microenvironment include TIE2-expressing monocytes, TIE2-expressing macrophages and TIE2-expressing endothelial cells.

Tumors that are responsive to angiopoietin / TIE2 signaling include, but are not limited to, breast cancer, ovarian cancer, hepatocellular carcinoma, glioma, colon cancer, and hematological malignancies.

を有する、遊離塩基性化合物である１−（３−ｔｅｒｔ−ブチル−１−（キノリン−６−イル）−１Ｈ−ピラゾール−５−イル）−３−（２−フルオロ−４−（２−（メチルカルバモイル）ピリジン−４−イルオキシ）フェニル）尿素である。 In another embodiment, in the absence of HX, the composition of formula I is: chemical structure:

1- (3-tert-Butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (2-( Methylcarbamoyl) pyridine-4-yloxy) phenyl) urea.

The composition of Formula I can be administered as a single agent or in combination with other therapeutic agents known to treat cancer. Other therapeutic agents such as these include radiation therapy, antitubulin agents, DNA alkylating agents, DNA synthesis inhibitors, DNA inserters, antiestrogen agents, antiandrogens, steroids, anti-EGFR agents, kinase inhibitors, topoisomerases. Inhibitors, histone deacetylase (HDAC) inhibitors, DNA methylation inhibitors, anti-HER2 agents, anti-angiogenic agents, proteasome inhibitors, salidamide, renalidemid, antibody-drug complex (ADC), immunomodulators or cancer A vaccine can be given.

Effective dose, toxicity and therapeutic efficacy are, for example, for determining LD50 (a dose lethal to about 50% of the population) and ED50 (a dose therapeutically effective in about 50% of the population). It can be determined by standard pharmaceutical techniques in cell culture or laboratory animals. The dose may vary depending on the dosage form used and the route of administration used. The dose ratio between toxicity and therapeutic effect is the therapeutic index and can be expressed as the LD50 / ED50 ratio. In some embodiments, compositions and methods that exhibit a large therapeutic index are preferred. A therapeutically effective dose can be first estimated from an in vitro assay, eg, a cell culture assay. Dose can also be formulated in an animal model that achieves a circulating plasma concentration range containing an IC50 determined in cell culture, or in a suitable animal model. The level of composition described in plasma can be measured, for example, by high performance liquid chromatography. The effect of any particular dose can be monitored by an appropriate bioassay. The dose will be determined by the physician and, if desired, may be adjusted to suit the observed effect of the procedure.

In certain embodiments, the prophylactic effect will result in a measurable change of at least about 10%, at least about 20%, at least about 30%, at least about 50%, at least about 70% or at least about 90%. In some embodiments, the effect will result in a quantitative change of about 10%, about 20%, about 30%, about 50%, about 70% or even about 90% or more. Therapeutic benefits also include stopping or delaying the progression of the underlying disease or disorder, whether or not improvement has been achieved.

When the composition of formula I or II is used in combination with another anti-cancer agent, the other anti-cancer agent may be administered independently of the dosing regimen of the composition of formula I or II. good. The other anti-cancer agent can be administered at its pre-established therapeutic dose and dosing regimen, or if that dose and dosing regimen is used in combination with the composition of formula I or II. Can be modified to optimize efficacy, safety or resistance.

In addition, any composition (and / or additional agent) of formula I or II described herein is administered to the subject as a component of a composition comprising a pharmaceutically acceptable carrier or vehicle. obtain. Such compositions may optionally contain appropriate amounts of pharmaceutically acceptable excipients to provide a suitable form for administration.

Pharmaceutical excipients can be liquids such as water and oils such as petroleum, animal, plant or synthetic origin such as peanut oil, soybean oil, mineral oil, sesame oil and the like. The pharmaceutical excipient may be, for example, physiological saline, gum arabic, gelatin, starch paste, talc, keratin, colloidal silica, urea and the like. In addition, auxiliaries, stabilizers, thickeners, lubricants and colorants can be used. In one embodiment, the pharmaceutically acceptable excipient is sterile at the time of administration to the subject. Water is a useful excipient when any of the agents described herein are administered intravenously. Aqueous saline solutions and aqueous dextrose and glycerol solutions can also be utilized as liquid excipients, especially for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, Examples include propylene glycol, water and ethanol. If desired, any of the agents described herein may also include small amounts of wetting or emulsifying agents or pH buffering agents.

The composition of Formula I may be used in combination with other agents, such as chemotherapeutic agents, targeted therapies, biopharmacy or radiation therapy.

The composition of the formula I is not limited to chemotherapeutic agents such as anti-tubulin agents (paclitaxel, paclitaxel protein binding particles for injection suspension, eribulin, docetaxel, ixabepyrone, vincristine), vinorelbine, DNA alkyl. Agents (including cisplatin, carboplatin, oxaliplatin, cyclophosphamide, iphosphamide, temozolomid), DNA inserts (including doxorubicin, pegulated liposomal doxorubicin, daunorubicin, idalubicin and epirubicin), 5-fluorouracil, capecitabine, It may be used in combination with cytarabine, decitabine, 5-azacitidine, gemcitabine and methotrexate.

The composition of the formula I is not limited to kinase inhibitors such as erlotinib, gefitinib, lapatinib, everolimus, temsirolimus, LY2835219, LEE011, PD0332991, crizotinib, cabozantinib, sunitinib, pazopanib, sorafenib, pazopanib, sorafenib, pazopanib, sorafenib. , Nilotinib, bemurafenib, dabrafenib, tramethinib, ideraricib and cabozantinib.

The composition of Formula I is not limited, and may be used in combination with antiestrogens such as tamoxifen, fulvestrant, anastrozole, letrozole and exemestane.

The composition of the formula I is not limited, and may be used in combination with an antiandrogen, for example, avirateron acetate, enzalutamide, nilutamide, bicalutamide, flutamide, cyproterone acetate.

The composition of Formula I is not limited, and may be used in combination with steroids such as prednisone and dexamethasone.

The composition of Formula I is not limited and may be used in combination with topoisomerase I inhibitors such as irinotecan, camptothecin and topotecan.

The composition of Formula I is not limited, and may be used in combination with topoisomerase II inhibitors, such as etoposide, etoposide phosphate and mitoxantrone.

The composition of formula I is not limited and may be used in combination with histone deacetylase (HDAC) inhibitors such as vorinostat, romidepsin, panobinostat, valproic acid and belinostat.

The composition of Formula I is not limited, and may be used in combination with DNA methylation inhibitors, such as DZNep and 5-aza-2'-deoxycytidine.

The composition of Formula I described above may be used in combination with proteasome inhibitors such as, but not limited to, vertezomib and carfilzomib.

The composition of Formula I may be used in combination with thalidomide, lenalidomide and pomalidomide.

The composition of the formula I is not limited to biologics such as trastuzumab, ado-trastuzumab, pertuzumab, cetuximab, panitumumab, ipilimumab, anti-PD-1 agents such as labrolizumab and nivolumab, anti-PD-L1 agents. For example, MPDL3280A, anti-angiogenic agents, such as bevacizumab and afribercept, and antibody-drug conjugates (ADCs), such as brentuximab bedotin and trastuzumab emtansine, may be used in combination.

The composition of Formula I may be used in combination with radiation therapy.

The composition of Formula I is not limited, and may be used in combination with a therapeutic vaccine, such as Sipuleucel-T.

In some embodiments, the composition of Formula I or Formula II can be used in combination with one or more of the other agents described herein.

Methods of Inhibiting the Growth and Invasion of Primary Breast Tumors The first aspect of the invention requires an effective amount of the composition of Formula I in a dosing regimen sufficient to inhibit TIE2 kinase in the tumor microenvironment. It relates to a method of inhibiting the growth and invasion of a primary breast tumor, including administration to a patient.

In one embodiment of this aspect of the invention, the dosing regimen for the composition of Formula I is daily dose administration.

In another embodiment of this aspect of the invention, the dosing regimen for the composition of Formula I is daily dose administration. Intermittent non-daily dosing regimens are not limited and may include dosing every other day, every two days, twice a week or once a week.

In another embodiment of this aspect of the invention, a suitable dosing regimen for the composition of Formula I comprises administration twice weekly, once weekly or once every two weeks.

In another embodiment of this aspect of the invention, the dosing regimen for the composition of Formula I is twice weekly or once weekly.

In another embodiment of this aspect of the invention, the dosing regimen for the composition of Formula I is administration twice weekly.

In yet another embodiment of the invention, the method of inhibiting the growth and invasion of the primary breast tumor is an anti-tubulin agent, a DNA alkylating agent, a DNA synthesis inhibitor, a DNA insert, an anti-estrogen agent, an anti-HER2. Administering the composition of formula I in a dosing regimen sufficient to inhibit TIE2 kinase in the tumor microenvironment in combination with one or more agents employed from agents, kinase inhibitors or anti-angiogenic agents. including.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administering the composition of formula I in combination with paclitaxel.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor administers the composition of formula I in combination with paclitaxel protein binding particles for an injectable suspension. Including to do.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administering the composition of formula I in combination with docetaxel.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of a primary breast tumor comprises administration of a composition of formula I in combination with eribulin.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of a primary breast tumor comprises administration of the composition of formula I in combination with ixabepyrone.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of a primary breast tumor comprises administration of the composition of formula I in combination with vinorelbine.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with capecitabine.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with gemcitabine.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with 5-fluorouracil.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with methotrexate.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with cyclophosphamide.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with cisplatin.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with carboplatin.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administering the composition of formula I in combination with doxorubicin.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administering the composition of formula I in combination with pegulated liposomal doxorubicin.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administering the composition of formula I in combination with epirubicin.

In yet another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with tamoxifen.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with fulvestrant.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with anastrozole.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with letrozole.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with exemestane.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with trastuzumab.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with ado-trastuzumab emtansine.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administering the composition of formula I in combination with pertuzumab.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with lapatinib.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with everolimus.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with temsirolimus.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of Formula I in combination with the CDK4 / 6 inhibitor LY2835219.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with the CDK4 / 6 inhibitor LEE011.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with the CDK4 / 6 inhibitor PD0332991.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with bevacizumab.

Another aspect of the invention comprises administration of the composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment, wherein the composition of Formula I is administered in an intermittent, non-daily dosing regimen. On how to inhibit the growth and invasion of primary breast tumors. In some embodiments, the intermittent non-daily dosing regimen includes every other day, every two days, twice a week, and once a week.

In another embodiment of this aspect of the invention, a method of inhibiting the growth and invasion of a primary breast tumor comprises administering the composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. , The composition of the formula I is administered twice a week, once a week or once every two weeks.

In another embodiment of this aspect of the invention, a method of inhibiting the growth and invasion of a primary breast tumor comprises administering the composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. , The composition of the formula I is administered twice a week or once a week.

In another embodiment of this aspect of the invention, a method of inhibiting the growth and invasion of a primary breast tumor comprises administering the composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. , The composition of formula I is administered twice a week.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of a primary estrogen tumor comprises administering the composition of formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. , The composition of the formula I is adopted from an anti-tubulin agent, a DNA alkylating agent, a DNA synthesis inhibitor, a DNA insertion agent, an anti-estrogen agent, an anti-HER2 agent, a kinase inhibitor or an anti-angiogenic agent. It is administered in combination with the above drugs.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administering the composition of formula I in combination with paclitaxel.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor administers the composition of formula I in combination with paclitaxel protein binding particles for an injectable suspension. Including to do.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administering the composition of formula I in combination with docetaxel.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of a primary breast tumor comprises administration of a composition of formula I in combination with eribulin.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of a primary breast tumor comprises administration of the composition of formula I in combination with ixabepyrone.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of a primary breast tumor comprises administration of the composition of formula I in combination with vinorelbine.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with capecitabine.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with gemcitabine.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with 5-fluorouracil.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with methotrexate.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with cyclophosphamide.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with cisplatin.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with carboplatin.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administering the composition of formula I in combination with doxorubicin.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administering the composition of formula I in combination with pegulated liposomal doxorubicin.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administering the composition of formula I in combination with epirubicin.

In yet another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with tamoxifen.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with fulvestrant.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with anastrozole.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with letrozole.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with exemestane.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with trastuzumab.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with ado-trastuzumab emtansine.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administering the composition of formula I in combination with pertuzumab.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of a primary breast tumor is to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with lapatinib. Includes administration of the composition of formula I in sufficient dose.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of a primary breast tumor is to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with everolimus. Includes administration of the composition of formula I in sufficient dose.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of a primary breast tumor is to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with temsirolimus. Includes administration of the composition of formula I in sufficient dose.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor is a TIE2 kinase in a tumor microenvironment TIE2-expressing macrophage administered in combination with the CDK4 / 6 inhibitor LY2835219. Includes administration of the composition of formula I at a dose sufficient to inhibit the.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor is a TIE2 kinase in a tumor microenvironment TIE2-expressing macrophage administered in combination with the CDK4 / 6 inhibitor LEE011. Includes administration of the composition of formula I at a dose sufficient to inhibit the.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor is a TIE2 kinase in a tumor microenvironment TIE2-expressing macrophage administered in combination with the CDK4 / 6 inhibitor PD0332991. Includes administration of the composition of formula I at a dose sufficient to inhibit the.

In another embodiment of this aspect of the invention, the method of inhibiting the growth and invasion of the primary breast tumor comprises administration of the composition of formula I in combination with bevacizumab.

Methods of Inhibiting Breast Cancer Invasion, Dissemination and Metastasis In yet another aspect of the invention, a patient requiring an effective amount of the composition of Formula I to inhibit TIE2 kinase in the tumor microenvironment. Provided are methods of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer, including administration to.

In one embodiment of this aspect of the invention, the method of inhibiting intravascular invasion, dissemination and metastasis of breast cancer is designed to adequately administer an effective amount of the composition of Formula I to inhibit TIE2 kinase in the tumor microenvironment. Including administration to a subject in need of it.

In another embodiment of this aspect of the invention, a dosing regimen sufficient to inhibit the intravascular invasion, dissemination and metastasis of the breast cancer comprises daily administration of the composition of formula I.

In another embodiment of this aspect of the invention, the dosing regimen of the composition of formula I is an intermittent non-daily dosing regimen, eg, every other day, every two days, twice a week or It is administered once a week.

In another embodiment of this aspect of the invention, the dosing regimen of the composition of formula I is administered twice weekly, once weekly or once every two weeks.

In another embodiment of this aspect of the invention, the dosing regimen for the composition of Formula I is administration twice weekly or once weekly.

In another embodiment of this aspect of the invention, the dosing regimen of the composition of formula I is administered twice weekly.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer is an antitubulin agent, a DNA kinase agent, a DNA synthesis inhibitor, a DNA inserter, an antiestrogen agent. In combination with one or more agents employed from anti-HER2 agents, kinase inhibitors or anti-angiogenic agents, the composition of an effective amount of Formula I is sufficient to inhibit TIE2 kinase in the tumor microenvironment. The dosing regimen includes administration to patients in need of it.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with paclitaxel.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer is in combination with paclitaxel protein-binding particles for an injectable suspension to formulate the composition of Formula I. Including administration.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with docetaxel.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with eribulin.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with ixabepyrone.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with vinorelbine.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with capecitabine.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with gemcitabine.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with 5-fluorouracil.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with methotrexate.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with cyclophosphamide.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with cisplatin.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with carboplatin.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with doxorubicin.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with epirubicin.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with tamoxifen.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with fulvestrant.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with anastrozole.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with letrozole.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with exemestane.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with trastuzumab.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with ado-trastuzumab emtansine.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with pertuzumab.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with lapatinib.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with everolimus.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with temsirolimus.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of Formula I in combination with the CDK4 / 6 inhibitor LY2835219.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of Formula I in combination with the CDK4 / 6 inhibitor LEE011.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of Formula I in combination with the CDK4 / 6 inhibitor PD0332991.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with bevacizumab.

In another aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment, said the formula. The dosing regimen of the composition of I is administered as an intermittent non-daily dosing. In some embodiments, the dosing every other day includes all dosing every two days, twice a week or once a week.

In one embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. The dosing regimen for the composition of Formula I is that it is administered twice a week, once a week or once every two weeks.

In another embodiment of this aspect of the invention, the method of inhibiting intravascular invasion, dissemination and metastasis of breast cancer involves administration of the composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. Including, the dosing regimen of the composition of formula I is subject to administration twice a week or once a week.

In another embodiment of this aspect of the invention, the method of inhibiting intravascular invasion, dissemination and metastasis of breast cancer involves administration of the composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. Including, the dosing regimen of the composition of formula I is subject to administration twice a week.

In another embodiment of this aspect of the invention, methods of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer are antitubulin agents, DNA alkylating agents, DNA synthesis inhibitors, DNA insertion agents, anti-estrogen agents, Composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment, administered in combination with one or more agents employed from anti-HER2 agents, kinase inhibitors or anti-angiogenic agents. Including administration of.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with paclitaxel.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer is in combination with paclitaxel protein-binding particles for an injectable suspension to formulate the composition of Formula I. Including administration.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with docetaxel.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with eribulin.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with ixabepyrone.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with vinorelbine.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with capecitabine.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with gemcitabine.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with 5-fluorouracil.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with methotrexate.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with cyclophosphamide.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with cisplatin.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with carboplatin.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with doxorubicin.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with pegulated liposomal doxorubicin.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with epirubicin.

In yet another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with tamoxifen.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with fulvestrant.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with anastrozole.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with letrozole.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with exemestane.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with trastuzumab.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with ado-trastuzumab emtansine.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administering the composition of formula I in combination with pertuzumab.

In another embodiment of this aspect of the invention, the method of inhibiting intravascular invasion, dissemination and metastasis of breast cancer inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with lapatinib. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of inhibiting intravascular invasion, dissemination and metastasis of breast cancer inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with everolimus. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of inhibiting intravascular invasion, dissemination and metastasis of breast cancer inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with temsirolimus. Contains administration of the composition of Formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer is TIE2 in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor LY2835219. Includes administration of the composition of formula I at a dose sufficient to inhibit the kinase.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer is TIE2 in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor LEE011. Includes administration of the composition of formula I at a dose sufficient to inhibit the kinase.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer is TIE2 in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor PD0332991. Includes administration of the composition of formula I at a dose sufficient to inhibit the kinase.

In another embodiment of this aspect of the invention, the method of inhibiting the intravascular invasion, dissemination and metastasis of breast cancer comprises administration of the composition of formula I in combination with bevacizumab.

Methods of Inhibiting Breast Cancer Immune Tolerance Another aspect of the invention relates to methods of inhibiting breast cancer immune tolerance. The method comprises administering an effective amount of the composition of Formula I to a patient in need thereof. In one embodiment, the salt dosing regimen is sufficient to inhibit TIE2 kinase in the tumor microenvironment that mediates immune tolerance.

In one embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer administers an effective amount of the composition of Formula I sufficient to inhibit TIE2 kinase in the tumor microenvironment that mediates immune tolerance. The plan involves administration to patients in need of it.

In one embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises daily administration of the composition of formula I.

In another embodiment of this aspect of the invention, the composition of Formula I is administered in an intermittent, non-daily manner. In some embodiments, the intermittent non-daily method comprises every other day, every two days, twice a week or once a week.

In another embodiment of this aspect of the invention, administration of the composition of formula I is twice weekly, once weekly or once every two weeks.

In another embodiment of this aspect of the invention, the composition of Formula I is administered twice weekly or once weekly.

In another embodiment of this aspect of the invention, the composition of Formula I is administered twice weekly.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immunotolerance comprises an anti-tubulin agent, a DNA alkylating agent, a DNA synthesis inhibitor, a DNA insert, an anti-estrogen agent, an anti-HER2 agent. In combination with one or more agents employed from kinase inhibitors, anti-angiogenic agents or immunomodulators, an effective amount of the composition of Formula I is administered sufficient to inhibit TIE2 kinase in the tumor microenvironment. The plan involves administration to patients in need of it.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with paclitaxel.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance comprises administering the composition of formula I in combination with paclitaxel protein binding particles for an injectable suspension. ..

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with docetaxel.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance comprises administration of a composition of formula I in combination with eribulin.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with ixabepyrone.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance comprises administration of a composition of formula I in combination with vinorelbine.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with capecitabine.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with gemcitabine.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with 5-fluorouracil.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administration of a composition of formula I in combination with methotrexate.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with cyclophosphamide.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance comprises administration of a composition of formula I in combination with cisplatin.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administration of a composition of formula I in combination with carboplatin.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with doxorubicin.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with pegged liposomal doxorubicin.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with epirubicin.

In yet another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with tamoxifen.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with fulvestrant.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administration of a composition of formula I in combination with anastrozole.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with letrozole.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with exemestane.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with trastuzumab.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administration of a composition of formula I in combination with ado-trastuzumab emtansine.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with pertuzumab.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance is administered in combination with lapatinib at a dose sufficient to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages. Includes administration of the composition of formula I of.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance is administered in combination with everolimus at a dose sufficient to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages. Includes administration of the composition of formula I of.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance is administered in combination with temsirolimus at a dose sufficient to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages. Includes administration of the composition of formula I of.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor LY2835219. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor LEE011. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor PD0332991. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administration of a composition of formula I in combination with bevacizumab.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with an anti-CTLA-4 agent.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administering the composition of formula I in combination with ipilimumab.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of Formula I in combination with an anti-PD-1 agent.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with lambbrorizumab.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with an anti-PD L-1 agent.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with MPDL3280A.

Methods of Inhibiting Breast Cancer Immune Tolerance In another aspect of the invention, methods of inhibiting breast cancer tolerance include administration of the composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. , The dosing regimen of the composition of formula I is administered by intermittent non-daily administration methods, such as every other day, every two days, twice a week or once a week.

In one embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering a composition of formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment, wherein the formula I. The composition is administered twice weekly, once weekly or once every two weeks.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment, said Formula I. The composition of is administered twice a week or once a week.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment, said Formula I. The composition of is administered twice a week.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immunotolerance is an anti-tubulin agent, a DNA alkylating agent, a DNA synthesis inhibitor, a DNA inserter, an anti-estrogen agent, an anti-HER2 agent, a kinase. Of the composition of formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment, administered in combination with one or more agents employed from inhibitors, anti-angiogenic agents or immunomodulators. Including administration.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with paclitaxel.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance comprises administering the composition of formula I in combination with paclitaxel protein binding particles for an injectable suspension. ..

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with docetaxel.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance comprises administration of a composition of formula I in combination with eribulin.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with ixabepyrone.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance comprises administration of a composition of formula I in combination with vinorelbine.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with capecitabine.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with gemcitabine.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with 5-fluorouracil.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administration of a composition of formula I in combination with methotrexate.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with cyclophosphamide.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance comprises administration of a composition of formula I in combination with cisplatin.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administration of a composition of formula I in combination with carboplatin.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with doxorubicin.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with pegged liposomal doxorubicin.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with epirubicin.

In yet another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with tamoxifen.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with fulvestrant.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administration of a composition of formula I in combination with anastrozole.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with letrozole.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with exemestane.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administration of a composition of formula I in combination with trastuzumab.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administration of a composition of formula I in combination with ado-trastuzumab emtansine.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with pertuzumab.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance is administered in combination with lapatinib at a dose sufficient to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages. Includes administration of the composition of formula I of.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance is administered in combination with everolimus at a dose sufficient to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages. Includes administration of the composition of formula I of.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance is administered in combination with temsirolimus at a dose sufficient to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages. Includes administration of the composition of formula I of.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor LY2835219. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor LEE011. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer tolerance inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor PD0332991. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administration of a composition of formula I in combination with bevacizumab.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with an anti-CTLA-4 agent.

In another embodiment of this aspect of the invention, the method of inhibiting immune tolerance of breast cancer comprises administering the composition of formula I in combination with ipilimumab.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of Formula I in combination with an anti-PD-1 agent.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with lambbrorizumab.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with an anti-PD L-1 agent.

In another embodiment of this aspect of the invention, the method of inhibiting breast cancer immune tolerance comprises administering the composition of formula I in combination with MPDL3280A.

Methods of Extending Overall Survival in Breast Cancer Patients Another aspect of the invention relates to methods of extending overall survival in breast cancer patients, comprising administering an effective amount of the composition of Formula I to a patient in need thereof. In one embodiment, the dosing regimen is sufficient to inhibit TIE2 kinase in the tumor microenvironment.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises a dosing regimen in which the composition of Formula I is administered daily.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient is an intermittent non-daily method, eg, every other day, every two days, twice a week or Includes the composition of formula I in a dosing regimen administered in weekly doses.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises the composition of formula I in a dosing regimen administered twice weekly, once weekly or once every two weeks.

In another embodiment of this aspect of the invention, in a method of prolonging overall survival in the breast cancer patient, the dosing regimen of the composition of formula I is administered twice weekly or once weekly.

In another embodiment of this aspect of the invention, the dosing regimen of the composition of formula I is administered only twice a week.

Another embodiment of this aspect of the invention is an anti-tubulin agent, a DNA alkylating agent, a DNA synthesis inhibitor, a DNA inserter, an anti-estrogen agent, an anti-HER2 agent, a kinase inhibitor, an anti-angiogenic agent or an immunomodification. Overall survival in breast cancer patients, including administration of the composition of Formula I in combination with one or more agents adopted from the agent in a dosing regimen sufficient to inhibit TIE2 kinase in the tumor microenvironment. Regarding how to stretch.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with paclitaxel.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with paclitaxel protein binding particles for an injectable suspension. ..

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with docetaxel.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with eribulin.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with ixabepyrone.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with vinorelbine.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with capecitabine.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with gemcitabine.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with 5-fluorouracil.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with methotrexate.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with cyclophosphamide.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with cisplatin.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with carboplatin.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with doxorubicin.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with pegged liposomal doxorubicin.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with epirubicin.

In yet another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with tamoxifen.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with fulvestrant.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with anastrozole.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with letrozole.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with exemestane.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with trastuzumab.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with ado-trastuzumab emtansine.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with pertuzumab.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient is administered in combination with lapatinib at a dose sufficient to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages. Includes administration of the composition of formula I of.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient is administered in combination with everolimus at a dose sufficient to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages. Includes administration of the composition of formula I of.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient is administered in combination with temsirolimus at a dose sufficient to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages. Includes administration of the composition of formula I of.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor LY2835219. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor LEE011. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor PD0332991. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with bevacizumab.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with an anti-CTLA-4 agent.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with ipilimumab.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with an anti-PD-1 agent.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with rambrolizumab.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with an anti-PD L-1 agent. In another embodiment of this aspect of the invention, the method of extending overall survival in a breast cancer patient comprises administering the composition of formula I in combination with MPDL3280A.

Methods to Extend Overall Survival in Breast Cancer Patients In another aspect of the invention, methods to increase overall survival in breast cancer patients include administration of the composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. The dosing regimen of the composition of formula I is intermittent non-daily administration, eg, every other day, every two days, twice a week or once a week.

In one embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment, wherein the formula I The dosing regimen of the composition is twice weekly, once weekly or once every two weeks.

In another embodiment of this aspect of the invention, a method of prolonging overall survival in a breast cancer patient comprises administering the composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment, said Formula I. The dosing regimen for the composition is twice weekly or once weekly.

In another embodiment of this aspect of the invention, a method of prolonging overall survival in a breast cancer patient comprises administering the composition of Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment, said Formula I. The dosing regimen for the composition of is the condition that it is administered twice a week.

In another embodiment of this aspect of the invention, methods of extending overall survival in breast cancer patients include antitubulin agents, DNA alkylating agents, DNA synthesis inhibitors, DNA inserters, antiestrogens, anti-HER2 agents, kinases. Of the composition of formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment, administered in combination with one or more agents employed from inhibitors, anti-angiogenic agents or immunomodulators. Including administration.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with paclitaxel.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with paclitaxel protein binding particles for an injectable suspension. ..

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with docetaxel.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with eribulin.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with ixabepyrone.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with vinorelbine.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with capecitabine.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with gemcitabine.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with 5-fluorouracil.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with methotrexate.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with cyclophosphamide.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with cisplatin.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with carboplatin.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with doxorubicin.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with pegged liposomal doxorubicin.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with epirubicin.

In yet another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with tamoxifen.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with fulvestrant.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with anastrozole.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with letrozole.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with exemestane.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with trastuzumab.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in the breast cancer patient comprises administration of the composition of formula I in combination with ado-trastuzumab emtansine.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with pertuzumab.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient is administered in combination with lapatinib at a dose sufficient to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages. Includes administration of the composition of formula I of.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient is administered in combination with everolimus at a dose sufficient to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages. Includes administration of the composition of formula I of.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient is administered in combination with temsirolimus at a dose sufficient to inhibit TIE2 kinase in tumor microenvironment TIE2-expressing macrophages. Includes administration of the composition of formula I of.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor LY2835219. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor LEE011. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages administered in combination with the CDK4 / 6 inhibitor PD0332991. Contains administration of the composition of formula I at a sufficient dose.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administration of a composition of formula I in combination with bevacizumab.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with an anti-CTLA-4 agent.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with ipilimumab.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with an anti-PD-1 agent.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with rambrolizumab.

In another embodiment of this aspect of the invention, the method of prolonging overall survival in a breast cancer patient comprises administering the composition of formula I in combination with an anti-PD L-1 agent.

In another embodiment of this aspect of the invention, the method of extending overall survival in a breast cancer patient comprises administering the composition of formula I in combination with MPDL3280A.

Methods of Treating Breast Cancer Patients in a Preoperative Auxiliary Environment Prior to Surgical Resection of Tumor Another aspect of the invention comprises administering to a patient in need of an effective amount of the composition of Formula I, said formula. The dosing regimen of the composition of I relates to a method of treating a breast cancer patient in a preoperative adjuvant environment prior to surgical resection of the tumor, where the dosing regimen is sufficient to inhibit TIE2 kinase in the tumor microenvironment.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of a tumor inhibits the composition of an effective amount of Formula I and TIE2 kinase in the tumor microenvironment. In a dosing regimen sufficient to include administration to patients in need of it.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is formulated I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. The dosing regimen of Formula I is provided daily, comprising administration of the composition of.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor inhibits the composition of an effective amount of Formula I and TIE2 kinase in the tumor microenvironment. In a dosing regimen sufficient to do this, the dosing regimen of formula I comprises administering to a patient in need thereof in an intermittent, non-daily manner, eg, every other day, every two days. , Twice a week or once a week.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises the composition of an effective amount of Formula I twice weekly, once weekly or A dosing regimen that is administered once every two weeks includes administration to patients in need of it.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is Formula I in a dosing regimen administered twice weekly or once weekly. Includes administration of the composition of.

In another embodiment of this aspect of the invention, the dosing regimen of the composition of formula I is administered twice weekly.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a preoperative assistive environment prior to surgical resection of the tumor is an antitubulin agent, a DNA kinase agent, a DNA synthesis inhibitor, a DNA insertion. Inhibits TIE2 kinase in tumor microenvironment TIE2-expressing macrophages in combination with one or more agents adopted from agents, anti-estrogen agents, anti-HER2 agents, kinase inhibitors, anti-angiogenic agents or immunomodulators. Includes administration of the composition of formula I with sufficient dosing regimen.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with paclitaxel.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is in combination with paclitaxel protein-binding particles for an injectable suspension. It involves administering the composition of formula I.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with docetaxel.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of a composition of formula I in combination with eribulin.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of a composition of formula I in combination with ixabepyrone.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with vinorelbine.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with capecitabine.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with gemcitabine.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with 5-fluorouracil.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with methotrexate.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of a composition of formula I in combination with cyclophosphamide. ..

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of a composition of formula I in combination with cisplatin.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of a composition of formula I in combination with carboplatin.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with doxorubicin.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor administers the composition of formula I in combination with pegulated liposomal doxorubicin. Including doing.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with epirubicin.

In yet another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with tamoxifen.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of a composition of formula I in combination with fulvestrant.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with anastrozole.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with letrozole.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with exemestane.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with trastuzumab.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is to administer the composition of formula I in combination with ado-trastuzumab emtansine. include.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with pertuzumab.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is TIE2 in tumor microenvironment TIE2-expressing macrophages administered in combination with lapatinib. Includes administration of the composition of formula I at a dose sufficient to inhibit the kinase.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is TIE2 in a tumor microenvironment TIE2-expressing macrophages administered in combination with everolimus. Includes administration of the composition of formula I at a dose sufficient to inhibit the kinase.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is TIE2 in a tumor microenvironment TIE2-expressing macrophages administered in combination with temsirolimus. Includes administration of the composition of formula I at a dose sufficient to inhibit the kinase.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is administered in combination with the CDK4 / 6 inhibitor LY283521, tumor microenvironment. Includes administration of the composition of formula I at a dose sufficient to inhibit TIE2 kinase in TIE2-expressing macrophages.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is administered in combination with the CDK4 / 6 inhibitor LEE011, a tumor microenvironment. Includes administration of the composition of formula I at a dose sufficient to inhibit TIE2 kinase in TIE2-expressing macrophages.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is administered in combination with the CDK4 / 6 inhibitor PD0332991, a tumor microenvironment. Includes administration of the composition of formula I at a dose sufficient to inhibit TIE2 kinase in TIE2-expressing macrophages.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with bevacizumab.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor administers the composition of formula I in combination with an anti-CTLA-4 agent. Including that.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with ipilimumab.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor administers the composition of formula I in combination with an anti-PD-1 agent. Including that.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with rambrolizumab.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor administers the composition of formula I in combination with an anti-PD L-1 agent. Including doing.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with MPDL3280A.

Methods of Treating Breast Cancer Patients in a Neoadjuvant Environment Before Surgical Resection In another aspect of the invention, methods of treating breast cancer patients in a neoadjuvant environment prior to surgical resection inhibit TIE2 kinase in the tumor microenvironment. The dosing regimen of the composition of Formula I comprises administration of the composition of Formula I in a dose sufficient to provide, for example, intermittent, non-daily administration, eg, every other day, every two days, weekly. The condition is that it is administered twice or once a week.

In one embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection is the composition of formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. The dosing regimen for the composition of Formula I, including administration, is that it is administered twice weekly, once weekly or once every two weeks.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection composes Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. The dosing regimen of the composition of formula I is that it is administered twice a week or once a week.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection composes Formula I at a dose sufficient to inhibit TIE2 kinase in the tumor microenvironment. The dosing regimen for the composition of Formula I is that it is administered twice a week.

In another embodiment of this aspect of the invention, methods of treating breast cancer patients in a preoperative assistive environment prior to surgical resection include anti-tubulin agents, DNA alkylating agents, DNA synthesis inhibitors, DNA insertion agents, anti-operatives. Sufficient to inhibit TIE2 kinase in tumor microenvironment, administered in combination with one or more agents adopted from estrogen, anti-HER2, kinase inhibitors, anti-angiogenic agents or immunomodulators Includes administration of the composition of formula I at a dose.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with paclitaxel.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is in combination with paclitaxel protein-binding particles for an injectable suspension. It involves administering the composition of formula I.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with docetaxel.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of a composition of formula I in combination with eribulin.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of a composition of formula I in combination with ixabepyrone.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with vinorelbine.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with capecitabine.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with gemcitabine.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of Formula I in combination with 5-fluorouracil.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with methotrexate.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of a composition of formula I in combination with cyclophosphamide. ..

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of a composition of formula I in combination with cisplatin.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of a composition of formula I in combination with carboplatin.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with doxorubicin.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor administers the composition of formula I in combination with pegulated liposomal doxorubicin. Including doing.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with epirubicin.

In yet another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with tamoxifen.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of a composition of formula I in combination with fulvestrant.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with anastrozole.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with letrozole.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with exemestane.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with trastuzumab.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is to administer the composition of formula I in combination with ado-trastuzumab emtansine. include.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with pertuzumab.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is TIE2 in tumor microenvironment TIE2-expressing macrophages administered in combination with lapatinib. Includes administration of the composition of formula I at a dose sufficient to inhibit the kinase.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is TIE2 in a tumor microenvironment TIE2-expressing macrophages administered in combination with everolimus. Includes administration of the composition of formula I at a dose sufficient to inhibit the kinase.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is TIE2 in a tumor microenvironment TIE2-expressing macrophages administered in combination with temsirolimus. Includes administration of the composition of formula I at a dose sufficient to inhibit the kinase.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is administered in combination with the CDK4 / 6 inhibitor LY283521, tumor microenvironment. Includes administration of the composition of formula I at a dose sufficient to inhibit TIE2 kinase in TIE2-expressing macrophages.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is administered in combination with the CDK4 / 6 inhibitor LEE011, a tumor microenvironment. Includes administration of the composition of formula I at a dose sufficient to inhibit TIE2 kinase in TIE2-expressing macrophages.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor is administered in combination with the CDK4 / 6 inhibitor PD0332991, a tumor microenvironment. Includes administration of the composition of formula I at a dose sufficient to inhibit TIE2 kinase in TIE2-expressing macrophages.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administration of the composition of formula I in combination with bevacizumab.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor administers the composition of formula I in combination with an anti-CTLA-4 agent. Including that.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with ipilimumab.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor administers the composition of formula I in combination with an anti-PD-1 agent. Including that.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with rambrolizumab.

In another embodiment of this aspect of the invention, the method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor administers the composition of formula I in combination with an anti-PD L-1 agent. Including doing.

In another embodiment of this aspect of the invention, a method of treating a breast cancer patient in a neoadjuvant environment prior to surgical resection of the tumor comprises administering the composition of formula I in combination with MPDL3280A.

Since TIE2 pathway signaling has been shown to be involved in the progression of ovarian cancer (Karlan et al, J. Clinical Oncology (2012) 30: 362-370), another aspect of the invention is to ovarian cancer. Regarding how to treat. The method comprises administering an effective amount of the composition of Formula I to a patient in need thereof in a dosing regimen sufficient to inhibit TIE2 kinase in the tumor microenvironment. In one embodiment of this aspect of the invention, the composition of formula I is administered as a single agent.

In another embodiment of this aspect of the invention, the composition of formula I is administered in a planned combination with paclitaxel + carboplatin, docetaxel + carboplatin, paclitaxel + cisplatin or other taxane + platinum drug.

Because TIE2 pathway signaling has been shown to be involved in the progression of hepatocellular carcinoma and has been shown as a diagnostic marker (Matsubara et al, Hepatology (2013) 57: 1416-1425; Mitsuhashi et al, Hepatology (Matsubara et al, Hepatology (2013)) 2003) 37: 1105-1113; Tanaka et al, J. Clin Invest (1999) 103: 341-345), another aspect of the invention relates to a method of treating hepatocellular carcinoma. The method comprises administering an effective amount of the composition of Formula I to a patient in need thereof in a dosing regimen sufficient to inhibit TIE2 kinase in the tumor microenvironment.

In one embodiment of this aspect of the invention, the composition of formula I is administered as a single agent.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with a kinase inhibitor such as sorafenib, crizotinib, cabozantinib, sunitinib, pazopanib, sorafenib, regorafenib or axitinib.

Because TIE2 pathway signaling has been shown to be involved in the progression of glioma cancer (Liu et al, Oncotarget (2010) 1: 700-709; Brunkhorst et al, Cancer Research (2010) 70: 7283-7293). , Another aspect of the invention relates to a method of treating a glioma. The method comprises administering an effective amount of the composition of Formula I to a patient in need thereof in a dosing regimen sufficient to inhibit TIE2 kinase in the tumor microenvironment.

In one embodiment of this aspect of the invention, the composition of formula I is administered as a single agent.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with radiation therapy.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with temozolomide therapy.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with radiation therapy and temozolomide therapy.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with bevacizumab therapy.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with radiation therapy and bevacizumab therapy.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with temozolomide therapy and bevacizumab therapy.

Because TIE2 pathway signaling has been shown to be involved in the progression of melanoma (Helfrich et al, Clin Cancer Res (2009) 15: 1384-1392; Peinado et al, Nature Medicine (2012) 18: 883-891) ), Another aspect of the invention relates to a method of treating melanoma. The method comprises administering an effective amount of the composition of Formula I to a patient in need thereof in a dosing regimen sufficient to inhibit TIE2 kinase in the tumor microenvironment.

In one embodiment of this aspect of the invention, the composition of formula I is administered as a single agent.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with vemurafenib.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with dabrafenib.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with dabrafenib and trametinib.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with temozolomide.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with dacarbazine.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with ipilimumab.

In another embodiment of this aspect of the invention, the composition of Formula I is administered in combination with labrolizumab or nivolumab.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with MPDL3280A.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with imatinib.

Because TIE2 pathway signaling has been shown to be involved in the progression of colorectal cancer (Ahmad et al, Cancer (2001) 92: 1138-1143; Hashizume et al, Cancer Research (2010) 70: 2213-2223). , Another aspect of the invention relates to a method of treating colorectal cancer. The method comprises administering an effective amount of the composition of Formula I to a patient in need thereof in a dosing regimen sufficient to inhibit TIE2 kinase in the tumor microenvironment.

In one embodiment of this aspect of the invention, the composition of formula I is administered as a single agent.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with mFOLFOX6 therapy (oxaliplatin + leucovorin + 5-fluorouracil).

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with mFOLFOX6 therapy and bevacizumab.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with mFOLFOX6 therapy and panitumumab.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with mFOLFOX6 therapy and cetuximab.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with capecitabine.

In another embodiment of this aspect of the invention, the composition of Formula I is administered in combination with capecitabine and bevacizumab.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with FOLFIRI therapy (irinotecan + leucovorin + 5-fluorouracil).

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with FOLFIRI therapy and bevacizumab.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with FOLFIRI therapy and aflibercept.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with FOLFIRI therapy and cetuximab.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with FOLFIRI therapy and panitumumab.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with panitumumab.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with panitumumab and irinotecan.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with cetuximab.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with cetuximab and irinotecan.

Because TIE2 pathway signaling has been shown to be involved in the progression of acute myeloid leukemia (Muller et al, Leukemia Research (2002) 26: 163-168; Hou et al, Leukemia Research (2008) 32:904). -912) Another aspect of the invention relates to a method of treating acute myeloid leukemia. The method comprises administering an effective amount of the composition of Formula I to a patient in need thereof in a dosing regimen sufficient to inhibit TIE2 kinase in the tumor microenvironment.

In one embodiment of this aspect of the invention, the composition of formula I is administered as a single agent.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with daunorubicin and cytarabine.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with idarubicin and cytarabine.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with mitoxantrone and cytarabine.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with cytarabine.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with 5-azacitidine.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with decitabine.

In another embodiment of this aspect of the invention, the composition of formula I is administered in combination with quizartinib.

Another aspect of the invention relates to a method of treating cancer. The method comprises administering an effective amount of the composition of Formula I to a patient in need thereof. In one embodiment, the patient overexpresses intimal endothelial cell kinase 2 (TIE2), and the cancers include breast cancer, colon cancer, hepatocellular carcinoma, head and neck cancer, bladder cancer, ovarian cancer, grioma. Select from hemangiosarcoma, melanoma or acute myeloid leukemia.

In certain embodiments of the treatment regimen of the present invention, the composition of Formula I is administered at a daily frequency.

In another embodiment of the treatment regimen of the present invention, the composition of Formula I is administered at a non-daily, intermittent frequency.

In another embodiment of the treatment regimen of the present invention, the composition of Formula I is administered at a frequency of 3 times a week.

In another embodiment of the treatment regimen of the present invention, the composition of Formula I is administered twice a week.

In another embodiment of the treatment regimen of the present invention, the composition of Formula I is administered at a frequency of once a week.

In another embodiment of the treatment regimen of the present invention, the composition of Formula I is administered at a frequency of once every two weeks.

In other embodiments, the cancer is a metastatic, three-negative breast cancer (estrogen receptor negative, progesterone receptor negative, HER2 negative).

In another embodiment, the cancer is estrogen-positive (ER ⁺⁾ and HER2 receptor kinase negative ^- is a breast cancer (HER2).

In another embodiment, the cancer is an inflammatory breast cancer.

In another embodiment, the method comprises a procedure that prevents or reduces the growth of one or more primary tumors, tumor invasion, cancer intravascular invasion, cancer dissemination, metastasis and tumor immune tolerance. include. In certain embodiments, the method improves patient survival.

Formulations, Administrations, Dose and Treatment Plans The present invention comprises the salts (and / or additional agents) of the formulas I and / or II described above in various formulations. Any composition (and / or additional agent) described herein is a capsule containing a liquid, a suspension, an emulsion, a drop, a tablet, a pill, a small pill, a capsule, a liquid. It may take the form of an agent, powder, sustained release formulation, suppository, emulsion, aerosol, spray, suspension, or any other form suitable for use. In one embodiment, the composition is in the form of a capsule (see, eg, US Pat. No. 5,698,155). Other examples of suitable pharmaceutical excipients are described in Remington's Pharmaceutical Sciences 1447-1676 (Alfonso R. Gennaro eds., 19th ed. 1995). This document is incorporated herein by reference.

If desired, the salts described herein may also include solubilizers. Also, the agent can be delivered by a suitable medium or delivery device known in the art. The combination therapies outlined herein can be delivered simultaneously in one delivery medium or device. The composition for administration may optionally include a local anesthetic, such as lignokine, to reduce pain at the injection site.

In one embodiment, the salts of formulas I and / or II (and / or additional agents) described herein are formulated according to conventional methods as compositions suitable for the method of administration.

In certain embodiments, the routes of administration include, for example, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, oral, sublingual, intranasal, intracerebral, intravaginal, transdermal. , Rectum, inhalation or topical to the ears, nose, eyes or skin in particular. In some embodiments, administration is achieved by oral or parenteral injection. The administration method can be at the discretion of the practitioner and can be determined to some extent depending on the site of the disease. In most cases, administration results in the release of any of the agents described herein into the blood.

In specific embodiments, it may be desirable to administer topically to areas requiring treatment or inhibition.

In one embodiment, the salts (and / or additional agents) described herein are formulated according to conventional methods as compositions suitable for oral administration to humans. Compositions for oral delivery can be, for example, in the form of tablets, lozenges, aqueous or oily suspensions, granules, powders, emulsions, capsules, syrups or elixirs. Orally administered compositions provide one or more agents, such as sweeteners, such as fructose, aspartame or saccharin; flavoring agents, such as peppermint, ichaxo, in order to provide a pharmaceutically palatable preparation. Alternatively, it may contain saccharin oil; colorant; and preservative. In addition, when in the form of tablets or pills, the composition can provide long-lasting effects by being coated to delay disintegration and absorption in the gastrointestinal tract. A selective permeable membrane environment that osmotically actively transports salts of formula I or II (and / or additional agents) described herein are also suitable for orally administered compositions. In these latter platforms, fluid from the environment surrounding the capsule is absorbed by the moving composition. The composition swells and moves the drug or drug composition through the pores. These delivery platforms may provide essentially zero order delivery profiles, as opposed to additional profiles of immediate release formulations. Time delay materials such as glycerol monostearate or glycerol stearate may also be useful. Oral compositions may include standard excipients such as mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose and magnesium carbonate. In one embodiment, the excipient is pharmaceutical grade. In addition to the active composition, the suspending agents include suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum hydroxide hydroxide, bentonite, agar-agar, tragacant and the like. And may include mixtures thereof.

Dosage forms suitable for parenteral administration (eg, intravenous, intramuscular, intraperitoneal, subcutaneous and intra-articular injections and infusions) include, for example, liquids, suspensions, dispersants, emulsions and the like. They can also be produced in the form of sterilized solid compositions (eg, lyophilized compositions). The composition may be dissolved or suspended in a sterile injectable medium immediately prior to use. They may include, for example, suspending agents or dispersants known in the art.

The doses and dosing schedules of the salts of formulas I and / or II (and / or additional agents) described herein are various parameters, eg, unrestricted and treated disease, overall subject. It may be determined by good health and the discretion of the administering physician. Any of the agents described herein is prior to administration of a further therapeutic agent to a subject in need thereof (eg, 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4). Hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks or 12 weeks ago), at the same time as the same administration Or after the same administration (eg, 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week , 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks or 12 weeks later). In various embodiments, any of the agents described herein is 1 minute apart, 10 minutes apart, 30 minutes apart, less than 1 hour apart, 1 hour apart, 1 hour to 2 hours. Free, 2 to 3 hours, 3 to 4 hours, 4 to 5 hours, 5 to 6 hours, 6 to 7 hours, 7 to 8 hours , 8 to 9 hours apart, 9 to 10 hours apart, 10 to 11 hours apart, or 11 to 12 hours apart.

The doses of salts (and / or additional agents) of formulas I and / or II described herein include multiple factors, such as the severity of the symptoms, whether the symptoms should be treated or prevented. , And may depend on the age, weight and health of the subject to be treated. In addition, pharmacological genomics (effect of genotype on therapeutic pharmacodynamic, pharmacodynamic or efficacy profile) information for a particular subject may affect the dose used. In addition, the exact individual dose can be determined by various factors, such as the specific combination of drugs administered, the timing of administration, the route of administration, the nature of the formulation, the rate of excretion, the specific disease to be treated, the severity of the disorder. And can be adjusted somewhat depending on the anatomical location of the disorder. Some variation of the dose can be predicted.

Generally, when administered orally to mammals, the dose of any composition (and / or additional agent) of formula I described herein is 0.001 mg / kg / day to 100 mg / kg. / Day, 0.01 mg / kg / day to 50 mg / kg / day or 0.1 mg / kg / day-10 mg / kg / day. When administered orally to humans, the dose of any of the agents described herein is typically 0.001 mg to 1500 mg per day or 1 mg to 600 mg per day or 5 mg to 30 mg per day. In some embodiments, the dose of said salt (or drug) ranges from 57 mg to 1200 mg per day. In other embodiments, the dose of the agent or salt ranges from 100 mg to 200 mg per day.

For administration by parenteral injection of salts of formulas I and / or II (and / or additional agents) described herein, the dose is typically 0.1 mg to 250 mg per day, 1 mg per day. ~ 20 mg or 3 mg ~ 5 mg per day. Injections may be provided up to 4 times per day. Generally, when administered orally or parenterally, the dose of any of the agents described herein is typically 0.1 mg to 1500 mg per day or 0.5 mg to 10 mg per day or It is 0.5 mg to 5 mg per day. A dose of 3000 mg or less can be administered per day.

Administration of the salts (and / or additional agents) described herein can be independently 1 to 4 times per day. Specifically, the administration of the salt can be once a day when the administration plan for the salt is about 50 mg to 1500 mg. The daily dose suitable for the required preventive effect is 57-1200 mg / day. When administered twice daily, the appropriate dose is 100 mg to 200 mg of the salt. Administration of the salt does not have to be intermittent and daily. In particular, the salt may be administered 1 to 4 times per month, 1 to 6 times per year, or once every 2, 3, 4 or 5 years. In certain embodiments, administration of the salt is done weekly or once every two weeks. When administered weekly or once every two weeks, the appropriate salt dosing regimen is in the range of 50-200 mg per dose. For certain weekly or biweekly doses, the dose is 200-400 mg per dose. As yet another method of administration once every week or two weeks, 400 to 500 mg per administration, 500 to 600 mg per administration, 600 to 700 mg per administration, once. 700 to 800 mg per administration, 800 to 900 mg per administration, 900 to 1000 mg per administration, 1000 to 1100 mg per administration, or 1100 to 1200 mg per administration. .. Administration can be for a period of 1 day or 1 month, 2 months, 3 months, 6 months, 1 year, 2 years, 3 years and may be during the life of the subject. Chronic, long-term administration will be required in many cases. The dose may be administered as a single dose or may be divided into multiple doses. In general, the desired dose should be administered over a long period of time, usually at intervals set over a period of at least weeks or months, but for a longer period of months or years or more. Administration may be required.

Example 1.1- (3-tert-butyl-1- (kinase-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl) pyridine-) Biochemical inhibition of non-phosphorylated TIE2 (uTIE2) by 4-yloxy) phenyl) urea Biochemical assay for uTIE2 (SEQ ID NO: 1) Kinase of uTIE2 kinase activity by coupling with pyruvate kinase / lactate dehydrogenase system It was determined by following the production of ADP from the reaction (eg, Chinder et al. Chemistry (2000) 289: 1938-1942). In this assay, NADH oxidation, which _{reduced A 340 nm} , was continuously monitored by spectroscopic measurements. The reaction mixture (100 μL) contains TIE2 (SignalChem) (5.6 nM), BSA (0.004% (0.004%), in 90 mM Tris buffer, pH 7.5, containing 0.2% octyl-glycoside and 1% DMSO. w / v)), polyEY (1.5 mg / ml), MgCl ₂ (15 mM), DTT (0.5 mM), pyruvate kinase (4 units), lactate dehydrogenase (7 units), phosphoenolpyruvate (1 mM) Also contained NADH (0.28 mM) and ATP (1.5 mM). The inhibitory reaction was initiated by mixing the serially diluted test composition with the above reaction mixture. Absorption at 340 nm was continuously monitored at 30 ° C. for 6 hours on a plate reader (Biotek). The reaction rate was calculated using a time frame of 5 to 6 hours. % Inhibition was obtained by comparing the kinetics with the kinetics of the control (ie, not including the test composition). From a series of percent inhibition values determined in the concentration range of the inhibitor using the same conventional software as is implemented in GraphPad Prism software package, the _{IC 50} value was calculated. 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl)) pyridine having the above composition. -4-Iloxy) phenyl) urea para-toluenesulfonate (Compound I shown in the figure) showed an _{IC 50 value of 3.5 nM.}
UTIE2 protein used for screening (SEQ ID NO: 1)
QLKRANVQRRMAQAFQNVREEPAVQFNSGTLALNRKVKNNPDPTIYPVLDWNDIKFQDVIGEGNFGQVLKARIKKDGLRMDAAIKRMKEYASKDDHRDFAGELEVLCKLGHHPNIINLLGACEHRGYLYLAIEYAPHGNLLDFLRKSRVLETDPAFAIANSTASTLSSQQLLHFAADVARGMDYLSQKQFIHRDLAARNILVGENYVAKIADFGLSRGQEVYVKKTMGRLPVRWMAIESLNYSVYTTNSDVWSYGVLLWEIVSLGGTPYCGMTCAELYEKLPQGYRLEKPLNCDDEVYDLMRQCWREKPYERPSFAQILVSLNRMLEERKTYVNTTLYEKFTYAGIDCSAEEAA

Examples 2.1- (3-tert-butyl-1- (kinase-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl) pyridine-) Biochemical inhibition of phosphorylated TIE2 (pTIE2) by 4-yloxy) phenyl) urea Biochemical assay for pTIE2 (SEQ ID NO: 2) Kinase reaction of pTIE2 kinase activity by coupling with pyruvate kinase / lactate dehydrogenase system Determined by following ADP production from (eg, Kinser et al. Chemistry (2000) 289: 1938-1942). In this assay, NADH oxidation, which _{reduced A 340 nm} , was continuously monitored by spectroscopic measurements. The reaction mixture (100 μL) contains TIE2 (Life Technologies) (6 nM), BSA (0.004% (w)) in 90 mM Tris buffer, pH 7.5, containing 0.2% octyl-glycoside and 1% DMSO. / V)), polyEY (1.5 mg / ml _{), MgCl 2} (15 mM), DTT (0.5 mM), pyruvate kinase (4 units), lactate dehydrogenase (7 units), phosphoenolpyruvate (1 mM) and It contained NADH (0.28 mM) and ATP (1.5 mM). The inhibitory reaction was initiated by mixing the serially diluted test composition with the above reaction mixture. Absorption at 340 nm was continuously monitored at 30 ° C. for 6 hours on a plate reader (Biotek). The kinetics were calculated using a 2-3 hour time frame. % Inhibition was obtained by comparing the kinetics with the kinetics of the control (ie, not including the test composition). From a series of percent inhibition values determined in the concentration range of the inhibitor using the same conventional software as is implemented in GraphPad Prism software package, the _{IC 50} value was calculated. 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl)) pyridine having the above composition. -4-Iloxy) phenyl) urea Para-toluene sulfonate and 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-) When 4- (2- (methylcarbamoyl) pyridin-4-yloxy) phenyl) urea bis-hydrochloride was tested, it showed more than 50% inhibition of pTIE2 kinase at concentrations below 0.1 μM. 1- (3-tert-Butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl) pyridin-4-yloxy) Phenyl) urea para-toluenesulfonate showed an _{IC 50 value of 4.2 nM.} 1- (3-tert-Butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl) pyridin-4-yloxy) phenyl) urea bis - hydrochloride exhibited an _{IC 50} value of 2.2 nM.
PTIE2 protein used for screening (SEQ ID NO: 2)
PVLDWNDIKFQDVIGEGNFGQVLKARIKKDGLRMDAAIKRMKEYASKDDHRDFAGELEVLCKLGHHPNIINLLGACEHRGYLYLAIEYAPHGNLLDFLRKSRVLETDPAFAIANSTASTLSSQQLLHFAADVARGMDYLSQKQFIHRDLAARNILVGENYVAKIADFGLSRGQEVYVKKTMGRLPVRWMAIESLNYSVYTTNSDVWSYGVLLWEIVSLGGTPYCGMTCAELYEKLPQGYRLEKPLNCDDEVYDLMRQCWREKPYERPSFAQILVSLNRMLEERKT

Example 3.1- (3-tert-Butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl) pyridine-) Cell inhibition of TIE2 in CHO cells by 4-yloxy) phenyl) urea Culture of CHO-K1 cells CHO-K1 cells (catalog # CCL-61) were obtained from the American Type Culture Collection (ATCC, Manassas, VA). rice field. Briefly in F12K medium supplemented with 10% charactified fetal bovine serum (Invitrogen, Carlsbad, CA), 100 units / mL penicillin G, 100 μg / ml streptomycin and 0.29 mg / mL L-glutamine (Invitrogen, Carlsbad, CA). Cells were grown at 37 ° C., 5% CO ₂ and 95% humidity. Cells were grown until they reached 70-95% confluence. At that time, the cells were subcultured or collected for use in the assay.

Phospho-TIE2 Western Blotting Assay of TIE2 Transformed CHO K1 CHO K1 cells (1 x 10 ⁵ cells / well) supplemented with 10% fetal bovine serum and 1 x non-essential amino acids (Invitrogen, Carlsbad, CA), 1 mL It was added to a 24-well tissue culture treatment plate in RPMI 1640 medium. The cells were then cultured overnight at 37 degrees Celsius, 5% CO ₂ and 95% humidity. The medium was aspirated and 0.5 mL of medium was added to each well. Transformation grade plasmid DNA (TIE2 gene gateway cloned into a cDNA 3.2 ™ / V5-DEST expression vector (Invitrogen, Carlsbad, CA)) at room temperature without serum Opti-MEM® It was diluted to 5 μg / mL in I medium (Invitrogen, Carlsbad, CA). 2 μL lipofectamine LTX reagent (Invitrogen, Carlsbad, CA) was added per 0.5 μg of plasmid DNA. The tubes were gently mixed and incubated at room temperature for 25 minutes to form a DNA-lipofectamine LTX complex. To each well containing the cells, 100 μL DNA-lipofectamine LTX complex was added directly and mixed gently. Twenty-four hours after transformation, medium containing the DNA-lipofectamine complex was aspirated. Cells were washed with PBS and RPMI 1640 medium supplemented with 10% charactified fetal bovine serum (Invitrogen, Carlsbad, CA) and 1 × non-essential amino acids (Invitrogen, Carlsbad, CA) was added. Test composition (1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl) pyridine-4) -Iloxy) phenyl) urea para-toluenesulfonate) or DMSO was added to the wells (0.5% final concentration of DMSO). The plate was then incubated at 37 degrees Celsius, 5% CO ₂ and 95% humidity for 4 hours. After incubation, the medium was aspirated and cells were washed with PBS. Shaking with an MPER lysis buffer (Pierce, Rockford, IL) containing Halt phosphatase and protease inhibitors (Pierce, Rockford, IL) and phosphatase inhibitor Cocktail 2 (Sigma, St. Louis, MO). The cells were lysed at 4 ° C. for 10 minutes. Clear lysates were separated by SDS-PAGE on a 4-12% Novex NuPage Bis-Tris gel (Invitrogen, Carlsbad, CA) and then transferred to PVDF (Invitrogen, Carlsbad, CA). After transcription, the PVDF membrane was blocked with BSA (Santa Cruz Biotechnology, Santa Cruz, CA) and then probed with an antibody for phospho-TIE2 (Cell Signaling Technology, Beverly, MA). A secondary anti-rabbit antibody (Cell Signaling Technology, Beverly, MA) conjugated to horseradish peroxidase was used to detect phospho-TIE2. ECL Plus (GE Healthcare, Piscataway, NJ), a substrate for horseradish peroxidase that produces fluorescence products, was added. Fluorescence was detected in fluorescence mode using a Storm 840 phosphor imager (GE Healthcare, Piscatawaym, NJ). The ImageQuant software (GE Healthcare, Piscatawaym, NJ) was used to quantify the 160 kDa phospho-TIE2 band. Prism software (GraphPad Software, San Diego, CA ) was used to analyze the data, the _{IC 50} value was calculated. 1- (3-tert-Butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl) pyridine-) having the above composition. 4-Iloxy) phenyl) urea para-toluenesulfonate showed an _{IC 50 value of 2.0 nM.}

Example 4.1- (3-tert-Butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl) pyridine-) Cell inhibition of TIE2 in CHO cells after washing with an inhibitor with 4-yloxy) phenyl) urea Culture of CHO-K1 cells CHO-K1 cells (catalog # CCL-61) were subjected to American Type Culture Collection (ATCC, Manassass). , VA). Briefly in F12K medium supplemented with 10% charactified fetal bovine serum (Invitrogen, Carlsbad, CA), 100 units / mL penicillin G, 100 μg / ml streptomycin and 0.29 mg / mL L-glutamine (Invitrogen, Carlsbad, CA). Cells were grown at 37 ° C., 5% CO ₂ and 95% humidity. Cells were grown until they reached 70-95% confluence. At that time, the cells were subcultured or collected for use in the assay.

Phosphor-TIE2 Western Blot Comparative Wash Assay for TIE2 Transformed CHO K1 CHO K1 cells (1 x 10 ⁵ cells / well) were supplemented with 10% charactified fetal bovine serum and 1 x non-essential amino acids (Invitrogen, Carlsbad, CA). It was added to a 24-well tissue culture treated plate in 1 mL RPMI 1640 medium. The cells were then cultured overnight at 37 degrees Celsius, 5% CO ₂ and 95% humidity. The medium was aspirated and 0.5 mL of medium was added to each well. Transformation grade plasmid DNA (TIE2 gene gateway cloned into a cDNA 3.2 ™ / V5-DEST expression vector (Invitrogen, Carlsbad, CA)) at room temperature without serum Opti-MEM® It was diluted to 5 μg / mL in I medium (Invitrogen, Carlsbad, CA). 2 μL lipofectamine LTX reagent (Invitrogen, Carlsbad, CA) was added per 0.5 μg of plasmid DNA. The tubes were gently mixed and incubated at room temperature for 25 minutes to form a DNA-lipofectamine LTX complex. To each well containing the cells, 100 μL DNA-lipofectamine LTX complex was added directly and mixed gently. Approximately 18-24 hours after transformation, medium containing the DNA-lipofectamine complex was aspirated. Cells were washed with PBS and RPMI 1640 medium supplemented with 10% charactified fetal bovine serum (Invitrogen, Carlsbad, CA) and 1 × non-essential amino acids (Invitrogen, Carlsbad, CA) was added. Test composition (1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl) pyridine-4) -Iloxy) phenyl) urea para-toluenesulfonate) or DMSO was added to the wells (DMSO final concentration 0.5%). The plate was then incubated at 37 degrees Celsius, 5% CO ₂ and 95% humidity for 2 hours. After incubation, the medium was aspirated and cells were washed 3 times with 1 mL of medium to wash away free composition. Next, 1 mL of fresh medium was added and the cells were incubated for a specified time (ie, 0, 1, 2, 4, 6 and 24 hours prior to lysis. Halt phosphatase and protease inhibitors (Pierce, Rockford, IL). ) And MPER lysis buffers (Pierce, Rockford, IL) containing phosphatase inhibitor cocktail 2 (Sigma, St. Louis, MO) were used to lyse the cells at 4 ° C. for 10 minutes with shaking. The clear protease was separated by SDS-PAGE in a 4-12% Novex NuPage Bis-Tris gel (Incubogen, Carlsbad, CA) and then transferred to PVDF (Invitrogen, Carlsbad, CA). After transfer, the PVDF. The membrane was blocked with BSA (Santa Cruz Biotechnology, Santa Cruz, CA) and then probed with an antibody for phospho-TIE2 (Cell Signaling Technology, Beverly, MA) and conjugated to Western wasabi peroxidase. Antibodies (Cell Signaling Technology, Beverly, MA) were used to detect phospho-TIE2. ECL Plus (GE Healthcare, Piscataway, NJ), a substrate for Western wasabi peroxidase that produces fluorescent products, was added. Fluorescence was detected in fluorescence mode using a Storm 840 phosphatase (GE Healthcare, Piscatawaym, NJ). The PVDF membrane was peeled off and, as described above, total TIE2 antibody (Santa Cruz Biotechnology, Inc.). , Dallas, TX). ImageQuant software (GE Healthcare, Piscatawaym, NJ) was used to quantify 160 kDa phospho-TIE2 and total TIE2 bands. Phospho-TIE2 levels relative to total TIE2 levels. Data were plotted using Prism software (GraphPad Software, San Diego, CA) when incubated with TIE2 transformed CHO K1 cells at 0.1 μM to 1 μM for 2 hours prior to washing. 1- (3-tert-Butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (2-( Methylcarbamoyl) pyridin-4-yloxy) phenyl) urea para-toluenesulfonate showed more than 50% inhibition of phospho-TIE2 levels over 24 hours.

Example 5.1- (3-tert-Butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl) pyridine-) Cell inhibition of TIE2 in HUVEC cells by 4-yloxy) phenyl) urea Culture of HUVEC cells HUVEC (human umbilical vein endothelial cells: Catalog # CRL-1730) cells from American Type Culture Collection (ATCC, Manassas, VA) Obtained. Briefly, cells were grown in EBM-2 (Lonza, Walkersville, MD) at 37 degrees Celsius, 5% CO ₂ and 95% humidity. Cells were grown until 90-95% saturation was reached. At that time, the cells were subcultured or collected for use in the assay.

HUVEC Phosphor-TIE2 Western Blot Assay HUVEC cells (2.5 × 10 ⁵ cells / well) were added to a 24-well tissue culture treatment plate in 1 mL EGM-2 culture medium (Lonza, Walkersville, MD). The cells were then cultured overnight at 37 degrees Celsius, 5% CO ₂ and 95% humidity. The medium was then aspirated and 1 mL of EBM-2 basal medium (Lonza, Walkersville, MD) supplemented with 2% FBS (Invitrogen, Carlsbad, CA) was added. The test composition or DMSO was added to the wells (0.5% final concentration of DMSO). The plate was then incubated at 37 degrees Celsius, 5% CO ₂ and 95% humidity for 4 hours. During the incubation, histidine-tagged angiopoietin 1 (ANG1) growth factors (R & D Systems, Minneapolis, MN) were added to anti-polyhistidine antibodies (R & D Systems, Minneapolis, MN) for 30 minutes at room temperature to add multimers of ANG1. Generated. After incubation with composition for 4 hours, cells were stimulated with 800 ng / mL ANG / anti-polyhistidine antibody complex mixture for 15 minutes. The medium was aspirated and the cells were washed with PBS. Shaking with an MPER lysis buffer (Pierce, Rockford, IL) containing Halt phosphatase and protease inhibitors (Pierce, Rockford, IL) and phosphatase inhibitor Cocktail 2 (Sigma, St. Louis, MO). The cells were lysed at 4 ° C. for 10 minutes. Clear lysates were separated by SDS-PAGE on a 4-12% Novex NuPage Bis-Tris gel (Invitrogen, Carlsbad, CA) and then transferred to PVDF (Invitrogen, Carlsbad, CA). After transcription, the PVDF membrane was blocked with BSA (Santa Cruz Biotechnology, Santa Cruz, CA) and then probed with an antibody for phospho-TIE2 (Cell Signaling Technology, Beverly, MA). A secondary anti-rabbit antibody (Cell Signaling Technology, Beverly, MA) conjugated to horseradish peroxidase was used to detect phospho-TIE2. ECL Plus (GE Healthcare, Piscataway, NJ), a substrate for horseradish peroxidase that produces fluorescence products, was added. Fluorescence was detected in fluorescence mode using a Storm 840 phosphor imager (GE Healthcare, Piscatawaym, NJ). ImageQuant software (GE Healthcare, Piscatawaym, NJ) was used to quantify the 160 kDa phospho-TIE2 band. Prism software (GraphPad Software, San Diego, CA ) was used to analyze the data, the _{IC 50} value was calculated. 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2), which is the composition disclosed in the present specification. -(Methylcarbamoyl) pyridin-4-yloxy) phenyl) urea para-toluenesulfonate showed an _{IC 50 value of 0.018 nM.}

Example 6.1- (3-tert-butyl-1- (quinolin-6-yl) -1H-pyrazol-5-yl) -3- (2-fluoro-4- (2- (methylcarbamoyl) pyridine-) Inhibition of angiopoietin 1 (ANG1) or angiopoietin 2 (ANG2) -stimulated capillary formation by 4-yloxy) phenyl) urea Culture of HMVEC cells HMVEC (human capillary endothelial cells: Catalog # PCS-110-010) cells, American Obtained from Type Culture Collection (ATCC, Manassas, VA). Briefly, cells were grown in EGM-2 MV (Lonza, Walkersville, MD) at 37 degrees Celsius, 5% CO ₂ and 95% humidity. Cells were grown until 90-95% saturation was reached. At that time, the cells were subcultured or collected for use in the assay.

HMVEC Capillary Angiogenesis Assay HMVEC cells (1.5 x 10 ⁴ cells / well) are mixed with the test composition or DMSO control and the appropriate growth factor (ANG1 or ANG2) or control is added to the 0.1 mL EBM-2 basis. It was added to 96-well tissue culture treated plates coated with growth factor-reduced Matrigel in medium (Lonza, Walkersville, MD). The cells were then cultured at 37 degrees Celsius, 5% CO ₂ and 95% humidity for 18 hours. The medium was then gently aspirated and the wells were gently washed with 0.1 mL EBM-2 basal medium. The medium was aspirated again and 1 μM Calcein AM solution (Invitrogen, Carlsbad, CA) in basal medium was added to each well to fluorescently label living cells. The cells were then incubated at 37 degrees Celsius, 5% CO ₂ and 95% humidity for 30 minutes. The medium was aspirated and the wells were gently washed twice with phosphate buffered saline. Images of each well were taken with a fluorescence microscope and processed using an ImagePro analyzer (Media Cybernetics, Inc., Rockville, MD) using an automated macro to measure the overall length of the capillaries. Prism software (GraphPad Software, San Diego, CA ) was used to analyze the data, the _{IC 50} value was calculated. 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2), which is the composition disclosed in the present specification. -(Methylcarbamoyl) pyridin-4-yloxy) phenyl) urea para-toluenesulfonate showed an _{IC 50 value of 6.9 nM for inhibition of ANG1-stimulated HMVEC capillary formation.} The composition of formula I as disclosed herein, for inhibition of ANG2 stimulation HMVEC capillary formation, exhibited _{IC 50} values 34 nM.

Example 7. 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2)) as a single agent and in combination with paclitaxel Inhibition of Invivo Primary Tumor Growth and Invasion in Mouse PyMT Breast Cancer Cells by − (Methylcarbamoyl) Pyridine-4-yloxy) Phenyl) Urea. It was used to evaluate the in vivo activity of Compound 1. ^{Briefly, 1 x 10 6} cells (separated from PyMT tumor fragments) in a total volume of 0.1 mL were separated from the fourth breast fat on the left side of a female mouse (FVB / NJ from Jackson Labs, JAXWEST: RB05 mouse). It was transplanted to the pad. In each group, a total of 10 mice were transplanted. Molecular Imaging, Inc. The Animal Care and Use Committee has approved all experimental protocols and conducted experiments in compliance with all regulations and guidelines of the National Institutes of Health (NIH). Based on oral administration (forced feeding) of compound 1 or vehicle (0.4% hydroxypropylmethylcellulose in water) twice daily and / or individual body weight on treatment day when tumor size reaches approximately 850 mg. Treatment was initiated by intravenous administration (IV) of paclitaxel or vehicle (10% ethanol, 10% Cremophor EL and 80% saline) every 4 days at 0.2 mL per 20 g. Animals were administered for 21 days. Body weight and tumor measurements were recorded 3 times a week. Tumor volume (mg) was estimated from caliper measurements by the formula for the amount of oblong ellipsoids assuming unit density: tumor mass (mg) = (L × W ² ) / 2. In the formula, L and W are measurements (mm) of each orthogonal tumor length and width. In the PyMT model, both Compound 1 and the paclitaxel group demonstrated tumor growth inhibition. Compound 1 in combination with paclitaxel demonstrated additive activity (Fig. 1). These data demonstrate in vivo activity by Compound 1 and show a correlation with enzyme and cellular data.

Example 8. 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2)) as a single agent and in combination with paclitaxel -Inhibition of in vivo primary tumor macrophages accumulation in mouse PyMT breast cancer model by − (methylcarbamoyl) pyridine-4-yloxy) phenyl) urea Macrophage accumulation of primary tumor in PyMT syngeneic breast cancer model It was used to evaluate the in vivo activity of Compound 1. ^{Briefly, 1 x 10 6} cells (separated from PyMT tumor fragments) in a total volume of 0.1 mL were separated from the fourth breast fat on the left side of a female mouse (FVB / NJ from Jackson Labs, JAXWEST: RB05 mouse). It was transplanted to the pad. In each group, a total of 10 mice were transplanted. Molecular Imaging, Inc. The Animal Care and Use Committee has approved all experimental protocols and conducted experiments in compliance with all regulations and guidelines of the National Institutes of Health (NIH). Based on oral administration (forced feeding) of compound 1 or vehicle (0.4% hydroxypropylmethylcellulose in water) twice daily and / or individual body weight on treatment day when tumor size reaches approximately 850 mg. Treatment was initiated by intravenous administration (IV) of paclitaxel or vehicle (10% ethanol, 10% Cremophor EL and 80% saline) every 4 days at 0.2 mL per 20 g. Animals were administered for 21 days. Body weight and tumor measurements were recorded 3 times a week.

At the end of the study, the tumor was resected and placed in formalin. The formalin-fixed tumor sample was then placed in a paraffin block. Formalin-fixed paraffin-embedded tissue slides were deparaffinized with xylene, hydrated, and water was evaporated by a series of stepwise alcohol washes. Antigen recovery was performed at 95 ° C. for 20 minutes using a Dako's PT Link Module containing a Tris / EDTA buffer target recovery solution. After cooling the slides, the slides were placed on Dako AutosteinerPlusLink for immunohistochemical staining using F4 / 80 and CD31 antibodies at room temperature and stained for macrophages and endothelial cells, respectively. Endogenous peroxidase and alkaline phosphatase activity in tissues was stopped for 5 minutes with a double endogenous enzyme inhibitory solution (Dako, S2003). Non-specific protein bindings were blocked with serum-free Protein Block (Dako, X0909) for 5 minutes. The rat anti-mouse CD31 primary antibody was then incubated at an immunogenicity concentration of 1: 100 on the experimental tissue section for 30 minutes. The primary antibody was then conjugated to a rabbit anti-rat immunoglobulin secondary antibody (Dako, E0468). The secondary antibody was then amplified with a goat anti-rabbit peroxidase-labeled polymer (Dako, K4003). Enzyme staining was performed on substrate-dye source DAB + (Dako, K3468) for 5 minutes. Excess rat IgG components were further blocked with Rodent Block Rat (Biocare Medical, RBR962H) for 5 minutes. The rat anti-mouse F4 / 80 primary antibody was incubated on the experimental tissue section for 30 minutes. The F4 / 80 was then conjugated with a rabbit anti-rat immunoglobulin secondary antibody (Dako, E0468). The secondary antibody was then amplified with a goat anti-rabbit alkaline phosphatase-labeled polymer (Biocare RALP525) for 30 minutes. Enzyme staining was performed on the substrate dye source WARP Red (Biocare WR806). Counter staining was performed with automatic hematoxylin for 10 minutes. The tissue slide was then air dried and passed through xylene to slide the slide glass. 0, no visible staining; 1, weak staining; 2, moderate staining; 3, strong staining scale was used to score slides for F4 / 80 staining. In the PyMT model, Compound 1 proved to reduce macrophage accumulation in said primary tumors. Paclitaxel, on the other hand, did not reduce macrophage accumulation. Compound 1 in combination with paclitaxel demonstrated activity similar to single-agent treatment of compound 1 (Fig. 2). These data demonstrate in vivo activity by Compound 1 and show a correlation with enzyme and cellular data.

Example 9. 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2)) as a single agent and in combination with paclitaxel Inhibition of TIE2 cell accumulation of in vivo primary breast cancer in mouse PyMT breast cancer model by − (methylcarbamoyl) pyridine-4-yloxy) phenyl) urea Macrophage accumulation of primary tumor in PyMT syngeneic breast cancer model PyMT syngeneic breast cancer transplanted mouse model , Used to assess the in vivo activity of Compound 1. ^{Briefly, 1 x 10 6} cells (separated from PyMT tumor fragments) in a total volume of 0.1 mL were separated from the fourth breast fat on the left side of a female mouse (FVB / NJ from Jackson Labs, JAXWEST: RB05 mouse). It was transplanted to the pad. In each group, a total of 10 mice were transplanted. Molecular Imaging, Inc. The Animal Care and Use Committee has approved all experimental protocols and conducted experiments in compliance with all regulations and guidelines of the National Institutes of Health (NIH). Based on oral administration (forced feeding) of compound 1 or vehicle (0.4% hydroxypropylmethylcellulose in water) twice daily and / or individual body weight on treatment day when tumor size reaches approximately 850 mg. Treatment was initiated by intravenous administration (IV) of paclitaxel or vehicle (10% ethanol, 10% Cremophor EL and 80% saline) every 4 days at 0.2 mL per 20 g. Animals were administered for 21 days. Body weight and tumor measurements were recorded 3 times a week.

At the end of the study, the tumor was resected and placed in formalin. The formalin-fixed tumor sample was then placed in a paraffin block. Experimental formalin-fixed paraffin-embedded tissue slides were deparaffinized with xylene, hydrated, and water was evaporated by a series of stepwise alcohol washes. Antigen recovery was performed at 95 ° C. for 20 minutes using a Dako's PT Link Module containing a citric acid pH 6 buffer target recovery solution. After cooling the slide, the slide was placed on a Dako AutosteinerPlusLink for immunohistochemical staining using TIE2 and CD31 antibodies at room temperature. The activity of endogenous peroxidase and alkaline phosphatase in tissues was stopped for 5 minutes with a double endogenous enzyme inhibitory solution (Dako, S2003). Non-specific protein bindings were blocked with serum-free Protein Block (Dako, X0909) for 5 minutes. The rat anti-mouse CD31 primary antibody was then incubated at an immunogenicity concentration of 1: 100 on the experimental tissue section for 30 minutes. The primary antibody was then conjugated to a rabbit anti-rat immunoglobulin secondary antibody (Dako, E0468). The secondary antibody was then amplified with a goat anti-rabbit peroxidase-labeled polymer (Dako, K4003) for 30 minutes. Enzyme staining was performed on substrate-dye source DAB + (Dako, K3468) for 5 minutes. Excess protein components were further blocked with Protein Block (Dako, X0909) for 5 minutes. The rabbit anti-TIE2 primary antibody was incubated on the experimental tissue section for 30 minutes. The TIE2 antibody was then conjugated to an alkaline phosphatase-labeled goat anti-rabbit polymer for 30 minutes. Enzyme staining was performed on the substrate dye source WARP Red (Biocare WR806). Counter staining was performed with automatic hematoxylin for 10 minutes. The tissue slide was then air dried and passed through xylene to slide the slide glass. Slides were scored for TIE2 staining using 0, no visible staining; 1, weak staining; 2, moderate staining; 3, strong staining scale. In the PyMT model, Compound 1 proved to reduce the accumulation of TIE2-expressing cells in the primary tumor. On the other hand, paclitaxel did not reduce the accumulation of TIE2-expressing cells. Compound 1 in combination with paclitaxel demonstrated improved activity compared to single agent treatment of compound 1 (FIG. 3). These data demonstrate in vivo activity by Compound 1 and show a correlation with enzyme and cellular data.

Example 10. 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro-4- (2)) as a single agent and in combination with paclitaxel Inhibition of in vivo lung metastasis in mouse PyMT breast cancer model by − (methylcarbamoyl) pyridine-4-yloxy) phenyl) urea Pulmonary metastasis evaluation of PyMT syngeneic breast cancer model Used to evaluate. ^{Briefly, 1 x 10 6} cells (separated from PyMT tumor fragments) in a total volume of 0.1 mL were separated from the fourth breast fat on the left side of a female mouse (FVB / NJ from Jackson Labs, JAXWEST: RB05 mouse). It was transplanted to the pad. In each group, a total of 10 mice were transplanted. Molecular Imaging, Inc. The Animal Care and Use Committee has approved all experimental protocols and conducted experiments in compliance with all regulations and guidelines of the National Institutes of Health (NIH). Based on oral administration (forced feeding) of compound 1 or vehicle (0.4% hydroxypropylmethylcellulose in water) twice daily and / or individual body weight on treatment day when tumor size reaches approximately 850 mg. Treatment was initiated by intravenous administration (IV) of paclitaxel or vehicle (10% ethanol, 10% Cremophor EL and 80% saline) every 4 days at 0.2 mL per 20 g. Animals were administered for 21 days. Body weight and tumor measurements were recorded 3 times a week.

At the end of the study, lung tissue was resected and placed in formalin. The formalin-fixed lung sample was then placed in a paraffin block. Each lung block had three slides, cut and stained with hematoxylin and eosin, with two levels per slide. Metastatic lung nodules were counted microscopically. In the PyTM model, both Compound 1 and paclitaxel demonstrated similar reductions in lung metastases. Compound 1 in combination with paclitaxel demonstrated additive activity compared to single agent treatment (Fig. 4). These data demonstrate in vivo activity by Compound 1 and show a correlation with enzyme and cellular data.

Example 11. 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro) administered intermittently (not daily) in combination with paclitaxel Inhibition of in vivo lung metastasis in mouse PyMT breast cancer model by -4- (2- (methylcarbamoyl) pyridine-4-yloxy) phenyl) Pulmonary metastasis evaluation of PyMT syngeneic breast cancer model PyMT syngeneic breast cancer transplanted mouse model, compound 1 Was used to assess the in vivo activity of. ^{Briefly, 1 × 10 6} cells (separated from PyMT tumor fragments and cryopreserved in cell cryomedia) in a total volume of 0.1 mL were subjected to female mice (FVB / NJ from Jackson Labs, JAXWEST: RB05 mice). ) Was transplanted to the 4th breast fat pad on the left side. In each group, a total of 3 mice were transplanted. Molecular Imaging, Inc. The Animal Care and Use Committee has approved all experimental protocols and conducted experiments in compliance with all regulations and guidelines of the National Institutes of Health (NIH). Oral administration (forced feeding) of compound 1 or vehicle (0.4% hydroxypropylmethylcellulose in water) twice weekly when tumor size reaches approximately 600 mg, and / or based on individual body weight on treatment day Treatment was initiated by intravenous administration (IV) of paclitaxel or vehicle (10% ethanol, 10% Cremophor EL and 80% saline) every 4 days at 0.2 mL / 20 g. Animals were administered for 12 days. Body weight and tumor measurements were recorded 3 times a week.

At the end of the study, lung tissue was resected and placed in formalin. The formalin-fixed lung sample was then placed in a paraffin block. Each lung block had three slides, cut and stained with hematoxylin and eosin, with two levels per slide. Metastatic lung nodules were counted microscopically. In the PyTM model, paclitaxel demonstrated a reduction in lung metastases. Compound 1 in combination with paclitaxel demonstrated additive activity compared to single agent treatment (FIG. 5). These data demonstrate in vivo activity by Compound 1 and show a correlation with enzyme and cellular data.

Example 12. 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro) administered intermittently (not daily) in combination with eribulin Inhibition of in vivo lung metastasis in mouse PyMT breast cancer model by -4- (2- (methylcarbamoyl) pyridin-4-yloxy) phenyl) urea Pulmonary metastasis evaluation of PyMT syngeneic breast cancer model Compound 1 Was used to assess the in vivo activity of. ^{Briefly, 1 × 10 6} cells (separated from PyMT tumor fragments and cryopreserved in cell cryomedia) in a total volume of 0.1 mL were subjected to female mice (FVB / NJ from Jackson Labs, JAXWEST: RB05 mice). ) Was transplanted to the 4th breast fat pad on the left side. In each group, a total of 3 mice were transplanted. Molecular Imaging, Inc. The Animal Care and Use Committee has approved all experimental protocols and conducted experiments in compliance with all regulations and guidelines of the National Institutes of Health (NIH). When tumor size reaches approximately 600 mg, oral administration of compound 1 or vehicle (0.4% hydroxypropylmethylcellulose in water) twice weekly (forced feeding) and / or based on individual body weight on treatment day Treatment was initiated by intravenous administration (IV) of eribulin or vehicle (80% saline) at 0.2 mL / 20 g three times weekly. Animals were administered for 12 days. Body weight and tumor measurements were recorded 3 times a week.

At the end of the study, lung tissue was resected and placed in formalin. The formalin-fixed lung sample was then placed in a paraffin block. Each lung block had three slides, cut and stained with hematoxylin and eosin, with two levels per slide. Metastatic lung nodules were counted microscopically. In the PyTM model, eribulin demonstrated a reduction (or improvement at low doses) in lung metastases. Compound 1 in combination with eribulin demonstrated additive activity compared to single agent treatment (Fig. 6). These data demonstrate in vivo activity by Compound 1 and show a correlation with enzyme and cellular data.

Example 13. 1- (3-tert-butyl-1- (quinoline-6-yl) -1H-pyrazole-5-yl) -3- (2-fluoro) administered intermittently (not daily) in combination with eribulin Prolongation of overall survival in mouse PyMT breast cancer model with -4- (2- (methylcarbamoyl) pyridine-4-yloxy) phenyl) Survival evaluation of PyMT allogeneic breast cancer model PyMT allogeneic breast cancer transplanted mouse model in vivo Used to assess activity. ^{Briefly, 1 × 10 6} cells (separated from PyMT tumor fragments and cryopreserved in cell cryomedia) in a total volume of 0.1 mL were subjected to female mice (FVB / NJ from Jackson Labs, JAXWEST: RB05 mice). ) Was transplanted to the 4th breast fat pad on the left side. In each group, a total of 10 mice were transplanted. Molecular Imaging, Inc. The Animal Care and Use Committee has approved all experimental protocols and conducted experiments in compliance with all regulations and guidelines of the National Institutes of Health (NIH). Oral administration (forced feeding) of Compound 1 or vehicle (0.4% hydroxypropylmethylcellulose in water) once or twice weekly when tumor size reaches approximately 850 mg, and / or individual body weight on treatment day. Treatment was initiated by intravenous administration (IV) of eribulin or vehicle (80% saline) three times weekly at 0.2 mL per 20 g. Then, 3 days after the start of the procedure, the tumor was resected. It was then administered to animals during the survival experiment. Body weight and tumor measurements were recorded 3 times a week. In the PyTM model, eribulin did not demonstrate prolongation of survival at 0.1 mg / kg. Compound 1 in combination with eribulin demonstrated a significant prolongation of survival (Fig. 7). These data demonstrate in vivo activity by Compound 1 and show a correlation with enzyme and cellular data.

The present invention is not limited in scope by the specific embodiments disclosed in the examples intended as illustrations of a few aspects of the invention. Any functionally equivalent embodiment is within the scope of the invention. Indeed, in addition to those shown and described herein, various improvements of the invention will be apparent to those skilled in the art and are intended to be within the appended claims. ..

Equivalents One of ordinary skill in the art will recognize or confirm a number of equivalents for a particular embodiment specifically described herein, without using anything other than normal trial and error. Would be able to. Such equivalents are intended to be included within the scope of the claims below.

Claims (12)

A pharmaceutical composition for treating breast cancer in patients in need of treatment.
The following structures: about 100 mg per day to about 200 mg per day:

A compound selected from the compounds having, and a pharmaceutically acceptable salt thereof,
A pharmaceutical composition comprising an effective amount of paclitaxel. The pharmaceutical composition according to claim 1 , wherein about 100 mg of the compound is administered to the patient per day. The pharmaceutical composition according to claim 1 , wherein about 200 mg of the compound is administered to the patient per day. The pharmaceutical composition according to any one of claims 1 to 3 , wherein the breast cancer is a three-kind negative breast cancer. The pharmaceutical composition according to any one of claims 1 to 3 , wherein the breast cancer is an inflammatory breast cancer. The pharmaceutical composition according to any one of claims 1 to 3 , wherein the breast cancer is HER2 receptor kinase negative. A pharmaceutical composition for treating breast cancer in patients in need of treatment.
The following structures: about 100 mg per day to about 200 mg per day:

A compound selected from the compounds having, and a pharmaceutically acceptable salt thereof,
A pharmaceutical composition comprising administering to the patient an effective amount of carboplatin. The pharmaceutical composition according to claim 7 , wherein about 100 mg of the compound is administered to the patient per day. The pharmaceutical composition according to claim 7 , wherein about 200 mg of the compound is administered to the patient per day. The pharmaceutical composition according to any one of claims 7 to 9 , wherein the breast cancer is a three-kind negative breast cancer. The pharmaceutical composition according to any one of claims 7 to 9 , wherein the breast cancer is an inflammatory breast cancer. The pharmaceutical composition according to any one of claims 7 to 9 , wherein the breast cancer is HER2 receptor kinase negative.

Images