Combination therapy for cancers with braf mutations

Abstract

The present disclosure provides combination therapies for treating a cancer having a BRAF mutation, the combination therapies comprising administering to a subject an effective amount of (a) an epidermal growth factor receptor (EGFR) inhibitor; (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor; (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor. Also provided are compositions and kits related to the combination therapies.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of International Application No. PCT / CN2018 / 082191, filed April 8, 2018, which is hereby incorporated by reference in its entirety. Technical Field

[0003] The present disclosure relates generally to combination therapies for treating or delaying the progression of cancers harboring a BRAF mutation, as well as related compositions and kits. Background Art

[0004] v-raf murine sarcoma viral oncogene homolog B1 (BRAF) is a potent activator of the mitogen-activated protein kinase (MAPK) / extracellular signal-regulated kinase (ERK) signaling pathway, which is involved in regulating cell proliferation, differentiation, and survival in response to extracellular signaling. Cantwell-Dorris et al. (2011) Mol. Cancer Ther. 10, 385. This pathway can be activated when extracellular ligands bind to epidermal growth factor receptor (EGFR), thereby initiating a cascade of ERK signaling through RAS GTPase. Once RAS is involved, it recruits and activates RAF (such as ARAF, BRAF, and CRAF), which phosphorylates MAPK / ERK kinase (MEK), thereby triggering the downstream ERK pathway. Oikonomou et al. (2014) Oncotarget 5, 11752.

[0005] BRAF mutations have been detected in a variety of cancers, including approximately 59% of malignant melanomas, approximately 45% of papillary thyroid cancers, approximately 18% of colorectal cancers (CRC), approximately 4% of ovarian cancers, approximately 2% of breast cancers, and approximately 3% of lung cancers, making it a potential therapeutic target. Davies et al. (2002) Nature 417, 949; Xing et al. (2005) Endocr. Relat. Cancer 12, 245. Mutations in BRAF most commonly occur at nucleotide 1976, resulting in a change at a valine residue (V600). Davies et al. (2002) Nature 417, 949. Mutated BRAF may cause constitutive activation of MAPK / ERK kinase (MEK), which in turn phosphorylates ERK and leads to target gene transcription, thereby promoting tumor cell growth and survival in the absence of any extracellular stimulation. Hall et al. (2014) Cancer Control 21, 221; Cantwell-Dorris et al. (2011) Mol. Cancer Ther. 10, 385.

[0006] Chemotherapy, even highly aggressive regimens, does not respond well to treatment of cancers with BRAF mutations. For example, in a clinical trial, patients with metastatic CRC received FOLFOXIRI plus bevacizumab (a highly aggressive regimen), but the survival of patients with BRAF-mutant CRC was less than half that of patients with wild-type tumors (median overall survival, 19.0 months compared to 41.7 months). Loupakis F et al. (2014) N Engl J Med. 371, 1609.

[0007] The U.S. Food and Drug Administration (FDA) has approved BRAF inhibitors, alone or in combination with MEK inhibitors, for the treatment of metastatic melanoma cancers with BRAF mutations. Larkin et al. (2014) N. Engl. J. Med. 371, 1867; Robert et al. (2015) N. Engl. J. Med. 372, 30. To date, melanoma cancers with BRAF mutations V600K or V600E are the only cancer types with BRAF mutations for which there is an approved targeted therapy. However, the duration of response is limited due to the development of acquired and adaptive resistance. Manzano et al. (2016) Ann. Transl. Med. 4, 237; Barras (2015) Biomarkers in Cancer 7, 9. Studies have shown that ERK can be reactivated by EGFR-mediated activation of RAS and CRAF, thereby contributing to cancer cell resistance to BRAF inhibitors. Barras (2015) Biomarkers in Cancer 7, 9. In addition, it has been reported that the use of BRAF inhibitors may lead to the development of secondary skin tumors due to paradoxical activation of the MAPK pathway in cells without BRAF mutations, but combining BRAF inhibitors with MEK inhibitors can reduce the incidence of BRAF inhibitor-induced skin tumors. Chapman et al. (2011) N. Engl. J. Med. 364, 2507; Robert et al. (2011) Curr. Opin. Oncol. 23, 177; Larkin et al. (2014) N. Engl. J. Med. 371, 1867. Recent and current clinical trials for cancers with BRAF mutations often involve the combination of BRAF inhibitors with other therapeutic drugs or drug candidates targeting different pathways (such as EGFR, MEK, PI3K) and cytotoxic chemotherapy. Morris et al. (2013) F1000 Prime Rep. 5, 11; Hyman DM et al. (2015) N. Engl. J. Med. 373, 726.

[0008] Therefore, there remains a need for robust therapies for treating cancers harboring BRAF mutations.

[0009] The disclosures of all publications, patents, patent applications, and published patent applications mentioned herein are hereby incorporated by reference in their entirety. Summary of the Invention

[0010] Provided herein are compositions comprising: (a) an epidermal growth factor receptor (EGFR) inhibitor; (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor; and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor; wherein the composition does not comprise a BRAF inhibitor. In some embodiments, the composition consists of: (a) an epidermal growth factor receptor (EGFR) inhibitor; (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor; and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor.

[0011] Also provided herein is a method for treating cancer or delaying its progression in a subject, comprising administering to the subject an effective amount of (a) an epidermal growth factor receptor (EGFR) inhibitor; (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor; and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor, wherein the subject suffers from cancer or is at risk of developing a cancer with a BRAF mutation. In some embodiments, the method does not include administering a BRAF inhibitor to the subject during administration of (a) an epidermal growth factor receptor (EGFR) inhibitor; (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor; and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor. In some embodiments, the method does not include administering to the subject during administration of (a) an epidermal growth factor receptor (EGFR) inhibitor; (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor; and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor an additional therapeutic agent.

[0012] Also provided herein is a method for treating cancer or delaying its progression in a subject, comprising administering an effective amount of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof to the subject, wherein the subject suffers from cancer or is at risk of developing a cancer with a BRAF mutation. In some embodiments, the method does not include administering a BRAF inhibitor to the subject during administration of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the method does not include administering an additional therapeutic agent to the subject during administration of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are administered in a composition. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are administered continuously to the subject. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are administered intermittently to the subject.

[0013] In some embodiments, osimertinib or a salt thereof is administered to a subject at a daily dose of about 20-160 mg of osimertinib. In some embodiments, cobimetinib or a salt thereof is administered to a subject at a daily dose of about 20-60 mg of cobimetinib. In some embodiments, palbociclib or a salt thereof is administered to a subject at a daily dose of about 75-125 mg of palbociclib. In some embodiments, the subject is a human. In some embodiments, osimertinib or a salt thereof is administered to a subject at a daily dose of about 0.5-3 mg / kg of osimertinib. In some embodiments, cobimetinib or a salt thereof is administered to a subject at a daily dose of about 0.25-1 mg / kg of cobimetinib. In some embodiments, palbociclib or a salt thereof is administered to a subject at a daily dose of about 1-2.5 mg / kg of palbociclib.

[0014] Also provided herein is a method for treating cancer or delaying its progress in a subject, comprising administering an effective amount of cetuximab, cobimetinib or its salt and palbociclib or its salt to the subject, wherein the subject suffers from cancer or is at risk of developing a cancer with a BRAF mutation. In some embodiments, the method does not include administering a BRAF inhibitor to the subject during administration of cetuximab, cobimetinib or its salt and palbociclib or its salt. In some embodiments, the method does not include administering another therapeutic agent to the subject during administration of cetuximab, cobimetinib or its salt and palbociclib or its salt. In some embodiments, cetuximab, cobimetinib or its salt and palbociclib or its salt are administered in a composition. In some embodiments, cetuximab, cobimetinib or its salt and palbociclib or its salt are continuously administered to the subject. In some embodiments, cetuximab, cobimetinib or its salt and palbociclib or its salt are intermittently administered to the subject.

[0015] In some embodiments, 400 mg / m2 is infused over 120 minutes. 2 , followed by weekly infusions of 250 mg / m2 over 60 minutes 2 In some embodiments, the maximum infusion rate is about 10 mL / min. In some embodiments, cobimetinib or a salt thereof is administered to a subject at a daily dose of about 20-60 mg of cobimetinib. In some embodiments, palbociclib or a salt thereof is administered to a subject at a daily dose of about 75-125 mg of palbociclib. In some embodiments, the daily dose is about 150-400 mg / m2 per subject. 2 In some embodiments, the subject is administered cetuximab at a weekly dose of about 0.25-10 mg / kg of cobimetinib. In some embodiments, the subject is administered cobimetinib or a salt thereof at a daily dose of about 5-30 mg / kg of palbociclib.

[0016] In some embodiments, the cancer has a BRAF V600 mutation or a BRAF D581D mutation. In some embodiments, the BRAF V600 mutation is a BRAF V600E, V600D, or V600K mutation. In some embodiments, the cancer is a malignant epithelial tumor or carcinoma. In some embodiments, the cancer is a cancer selected from one or more of the following: colon cancer, gastric cancer, lung cancer, breast cancer, pancreatic cancer, oral cancer, prostate cancer, reproductive cancer, rectal cancer, liver cancer, kidney cancer, papillary thyroid cancer, and ovarian cancer. In some embodiments, the cancer is colorectal cancer. In some embodiments, the colorectal cancer is stage IV colorectal cancer. In some embodiments, the colorectal cancer has a BRAF V600E or D581V mutation.

[0017] In some embodiments, the subject has received a BRAF inhibitor in a previous treatment cycle. In some embodiments, the subject has not received a BRAF inhibitor in a previous treatment cycle.

[0018] In some embodiments, the methods provided herein reduce average tumor volume. In some embodiments, the cancer cell growth reduction and / or cancer cell killing increase of the methods provided herein are compared with administering (a) epidermal growth factor receptor (EGFR) inhibitors alone; (b) mitogen-activated protein kinase (MEK) 1 / 2 inhibitors; and (c) cyclin-dependent kinase (CDK) 4 / 6 inhibitors by about 20-99%. In some embodiments, the cancer cell growth reduction and / or cancer cell killing increase of the methods provided herein are compared with administering osimertinib or its salt, cobimetinib or its salt, or palbociclib or its salt by about 20-99%. In some embodiments, the cancer cell growth reduction and / or cancer cell killing increase of the methods provided herein are compared with administering cetuximab, cobimetinib or its salt, or palbociclib or its salt by about 20-99%. In some embodiments, the method reduces tumor volume by about 20-95%. The method may include administering any composition or kit described herein.

[0019] On the other hand, provided herein are compositions comprising the following: (a) epidermal growth factor receptor (EGFR) inhibitors; (b) mitogen-activated protein kinase (MEK) 1 / 2 inhibitors; and (c) cyclin-dependent kinase (CDK) 4 / 6 inhibitors. In some embodiments, the composition comprises osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the composition does not comprise a BRAF inhibitor. In some embodiments, the composition consists of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier, excipient, adhesive, or diluent. In some embodiments, the composition is formulated for oral administration to a subject. In some embodiments, the subject suffers from cancer or is at risk of developing a cancer with a BRAF mutation.

[0020] On the other hand, provided herein are kits comprising the following: (a) an epidermal growth factor receptor (EGFR) inhibitor; (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor; and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor; wherein the kit does not comprise a BRAF inhibitor. In some embodiments, the kit consists of the following: (a) an epidermal growth factor receptor (EGFR) inhibitor; (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor; and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor.

[0021] In some embodiments, the kit comprises osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the kit does not comprise a BRAF inhibitor. In some embodiments, the kit consists of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the kit comprises a pharmaceutical composition comprising osimertinib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising cobimetinib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising palbociclib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, osimertinib or a salt thereof is formulated for oral administration to a subject. In some embodiments, cobimetinib or a salt thereof is formulated for oral administration to a subject. In some embodiments, palbociclib or a salt thereof is formulated for oral administration to a subject. In some embodiments, the subject has cancer or is at risk of developing cancer with a BRAF mutation. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are formulated in a composition. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are administered continuously to the subject. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are administered intermittently to the subject.

[0022] In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are formulated into a composition. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are formulated into separate compositions. In some embodiments, osimertinib or a salt thereof and cobimetinib or a salt thereof are formulated into a composition. In some embodiments, osimertinib or a salt thereof and palbociclib or a salt thereof are formulated into a composition. In some embodiments, cobimetinib or a salt thereof and palbociclib or a salt thereof are formulated into a composition. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are formulated into a liquid form. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are formulated into a solid form.

[0023] In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are administered as a combination. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are administered separately. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are administered simultaneously. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are administered continuously. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof are administered intermittently.

[0024] In some embodiments, the kit comprises cetuximab, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the kit does not comprise a BRAF inhibitor. In some embodiments, the kit consists of cetuximab, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the kit comprises a pharmaceutical composition comprising cetuximab and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising cobimetinib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising palbociclib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, cetuximab is formulated for administration to a subject via intravenous infusion. In some embodiments, cobimetinib or a salt thereof is formulated for oral administration to a subject. In some embodiments, palbociclib or a salt thereof is formulated for oral administration to a subject. In some embodiments, the subject has cancer or is at risk of developing a cancer with a BRAF mutation. In some embodiments, cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof are formulated in different compositions. In some embodiments, cobimetinib or a salt thereof and palbociclib or a salt thereof are formulated in a composition. In some embodiments, cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof are continuously administered to the subject. In some embodiments, cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof are intermittently administered to the subject. In some embodiments, cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof are administered to the subject at different dosing frequencies.

[0025] In some embodiments, cetuximab, cobimetinib or its salt and palbociclib or its salt are formulated into a composition. In some embodiments, cetuximab, cobimetinib or its salt and palbociclib or its salt are formulated into separate compositions. In some embodiments, cetuximab and cobimetinib or its salt are formulated into a composition. In some embodiments, cetuximab and palbociclib or its salt are formulated into a composition. In some embodiments, cobimetinib or its salt and palbociclib or its salt are formulated into a composition. In some embodiments, cetuximab, cobimetinib or its salt and palbociclib or its salt are formulated into a composition. In some embodiments, cetuximab, cobimetinib or its salt and palbociclib or its salt are formulated into a liquid form. In some embodiments, cetuximab, cobimetinib or its salt and palbociclib or its salt are formulated into a solid form.

[0026] In some embodiments, cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof are administered as a combination. In some embodiments, cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof are administered separately. In some embodiments, cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof are administered simultaneously. In some embodiments, cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof are administered continuously. In some embodiments, cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof are administered intermittently. In some embodiments, cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof are administered at different dosing frequencies.

[0027] In some embodiments, the kit includes a package insert containing instructions regarding the indications, usage, dosage, administration, contraindications, other drugs to be combined with the packaged product, and / or warnings concerning the use of such drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 Shown are representative images of stained tumor cells obtained by the high-content screening (HCS) platform.

[0029] Figure 2Depicts reduction in CRC (NYL132) volume after treatment with osimertinib, cobimetinib, and palbociclib. The graph shows the change in mean tumor volume over time during combination therapy: (filled circles) - untreated control; (filled triangles) - oxaliplatin 10 mg / kg and capecitabine 200 mg / kg daily (po) for 3 weeks; (filled diamonds) - osimertinib 5 mg / kg, cobimetinib 2.5 mg / kg, and palbociclib 10 mg / kg daily (po) for 3 weeks; (open squares) - osimertinib 7.5 mg / kg, cobimetinib 3.75 mg / kg, and palbociclib 15 mg / kg daily (po) for 3 weeks; and (open circles) - osimertinib 10 mg / kg, cobimetinib 5 mg / kg, and palbociclib 20 mg / kg daily (po) for 3 weeks. Dosage units mg / kg refer to the compound dose per kg of mouse body weight.

[0030] Figure 3 Depicts reduction in CRC (NYL132) volume after treatment with osimertinib, cobimetinib, and palbociclib. The graph shows tumor growth inhibition (TGI) over time during combination therapy: (filled triangles) - oxaliplatin 10 mg / kg and capecitabine 200 mg / kg daily (po) for 3 weeks; (filled diamonds) - osimertinib 5 mg / kg, cobimetinib 2.5 mg / kg, and palbociclib 10 mg / kg daily (po) for 3 weeks; (open squares) - osimertinib 7.5 mg / kg, cobimetinib 3.75 mg / kg, and palbociclib 15 mg / kg daily (po) for 3 weeks; and (open circles) - osimertinib 10 mg / kg, cobimetinib 5 mg / kg, and palbociclib 20 mg / kg daily (po) for 3 weeks. Dosage units mg / kg refer to the compound dose per kg of mouse body weight. TGI=(1-(Ti-T0) / (Vi-V0))*100%.

[0031] Figure 4 Depicted are body weights of mice (NYL132) during combination therapy: (filled circles) - untreated controls; (filled triangles) - oxaliplatin 10 mg / kg and capecitabine 200 mg / kg daily (po) for 3 weeks; (filled diamonds) - osimertinib 5 mg / kg, cobimetinib 2.5 mg / kg, and palbociclib 10 mg / kg daily (po) for 3 weeks; (open squares) - osimertinib 7.5 mg / kg, cobimetinib 3.75 mg / kg, and palbociclib 15 mg / kg daily (po) for 3 weeks; and (open circles) - osimertinib 10 mg / kg, cobimetinib 5 mg / kg, and palbociclib 20 mg / kg daily (po) for 3 weeks.

[0032] Figure 5 Depicted is the reduction in volume of CRC (NYL-GZ-082) following treatment with the combination therapy. The graph shows the change in mean tumor volume over time during the combination treatment: (solid triangles) - control, with no treatment for 14 days, cetuximab 1 mg weekly (ip), cobimetinib 5 mg / kg and palbociclib 20 mg / kg daily (po) for 21 days, followed by no treatment for 1 week; second cycle starting on day 43 (same treatment as the first cycle (first 4 weeks)); (solid squares) - osimertinib 10 mg / kg, cobimetinib 5 mg / kg and palbociclib 20 mg / kg daily (po) for 3 weeks, followed by no treatment for 1 week; second cycle from days 28 to 56, with the same treatment as the first cycle (first 4 weeks); (open circles) - cetuximab 1 mg weekly (ip), cobimetinib 5 mg / kg and palbociclib 20 mg / kg daily (po) for 21 days, followed by no treatment for 1 week; second cycle starting from days 28 to 56, with the same treatment as the first cycle (first 4 weeks). The dosage unit mg / kg refers to the dose of compound per kg of mouse body weight.

[0033] Figure 6 Figure 3 shows the body weight changes (%) of mice (NYL-GZ-082) during combination therapy: (filled triangles) - control, in which no treatment lasted for 14 days, cetuximab 1 mg weekly (ip), cobimetinib 5 mg / kg and palbociclib 20 mg / kg daily (po) for 21 days, followed by no treatment for 1 week; second cycle starting on day 43 (same treatment as the first cycle (first 4 weeks)); (filled squares) - osimertinib 10 mg / kg, cobimetinib 5 mg / kg and palbociclib 20 mg / kg daily (po) for 21 days, followed by no treatment for 1 week. mg / kg and palbociclib 20 mg / kg daily (po) for 3 weeks, followed by no treatment for 1 week; Days 28-56 are the second cycle, with the same treatment as the first cycle (first 4 weeks); (open circles) - Cetuximab 1 mg weekly (ip), cobimetinib 5 mg / kg, and palbociclib 20 mg / kg daily (po) for 21 days, followed by no treatment for 1 week; Days 28-56 are the second cycle, with the same treatment as the first cycle (first 4 weeks).

[0034] Figure 7Figure 3 depicts the reduction in CRC (NYL132) volume after treatment with the combination therapy. The graph shows the change in mean tumor volume over time during the combination therapy: (solid triangles) - untreated control; (solid squares) - cetuximab 1 mg weekly (ip), cobimetinib 5 mg / kg, and palbociclib 20 mg / kg daily (po) for 21 days, no treatment for 7 days, followed by a second cycle with the same treatment as the first cycle (first 4 weeks); (solid circles) - osimertinib 10 mg / kg, TAK-733 3 mg / kg, and palbociclib 20 mg / kg daily (po) for 21 days, no treatment for 7 days, followed by a second cycle with the same treatment as the first cycle; (solid diamonds) - cetuximab 1 mg weekly (ip), TAK-733 =(open triangles) - cetuximab 1 mg weekly (ip) for 21 days, no treatment for 7 days, cetuximab 1 mg weekly (ip) on days 28-49, no treatment for another 7 days, followed by cetuximab 1 mg weekly (ip), cobimetinib 5 mg / kg, and palbociclib 20 mg / kg daily (po) for another 21 days; (open circles) - cetuximab 1 mg weekly (ip), TAK-733 10 mg / kg, and palbociclib 20 mg / kg daily (po) for 21 days, no treatment for 7 days, followed by a second cycle with the same treatment as the first cycle; (open diamonds) - osimertinib 10 mg / kg, TAK-733 10 mg / kg and palbociclib 20 mg / kg daily (po) for 21 days, no treatment for 7 days, followed by a second cycle with the same treatment as the first cycle.

[0035] Figure 8Figure 2 depicts the reduction in CRC (HT-29) volume after treatment with the combination therapy. The graph shows the change in mean tumor volume over time during the combination therapy: (solid triangles) - control with no treatment for 28 days; (solid squares) - capecitabine 200 mg / kg daily (po) for 21 days; (solid circles) - cetuximab 1 mg weekly (ip), cobimetinib 5 mg / kg, and palbociclib 20 mg / kg daily (po) for 21 days; (solid diamonds) - cetuximab 1 mg weekly (ip), cobimetinib 5 mg / kg, and palbociclib 13.3 mg / kg daily (po) for 21 days; (open triangles) - cetuximab 1 mg weekly (ip), cobimetinib 5 mg / kg, and palbociclib 13.3 mg / kg daily (po) for 21 days. =Osimertinib 5 mg / kg and palbociclib 6.6 mg / kg daily (po) for 21 days; (open squares) - osimertinib 10 mg / kg, cobimetinib 5 mg / kg, and palbociclib 20 mg / kg daily (po) for 21 days; (open circles) - osimertinib 10 mg / kg and cobimetinib 5 mg / kg daily (po) for 21 days; (open diamonds) - cobimetinib 5 mg / kg daily (po) for 21 days; (dashed solid triangles) - cobimetinib 5 mg / kg and palbociclib 20 mg / kg daily (po) for 21 days; (dashed solid circles) - 5-FU 40 mg / kg weekly for 3 weeks. On day 21, all treatments were stopped, followed by 1 week of observation.

[0036] Figure 9Depicts the reduction in CRC (HT-29) volume after treatment with the combination therapy. The graph shows the change in mean tumor volume over time during the combination therapy: (solid triangles) - control without treatment for 21 days; (solid squares) - TAK-733 10 mg / kg daily (po) for 21 days; (solid circles) - TAK-733 on day 1 30 mg / kg, then 3 mg / kg daily (po) for 20 days; (filled diamonds) - palbociclib 20 mg / kg daily (po) for 21 days; (open triangles) - cetuximab 1 mg weekly (ip) for 21 days, followed by 7 days off treatment, then 1 mg weekly (ip) for another 14 days; (open squares) - capecitabine 200 mg / kg daily (po) and oxaliplatin 10 mg / kg weekly (ip) for 21 days, followed by 7 days off treatment; after day 28, cetuximab 1 mg weekly (ip), cobimetinib 5 mg / kg, and palbociclib 20 mg / kg daily (po) for 14 days; (open circles) - osimertinib 10 mg / kg, TAK-733 =(1) Cetuximab 1 mg weekly (ip), TAK-733 10 mg / kg, and palbociclib 20 mg / kg daily (po) for 21 days, followed by 7 days off treatment, followed by 14 days at the same dose; (open diamonds) - Cetuximab 1 mg weekly (ip), TAK-733 10 mg / kg, and palbociclib 20 mg / kg daily (po) for 21 days, followed by 7 days off treatment, followed by 14 days at the same dose; (dashed solid triangles) - TAK-733 10 mg / kg, and palbociclib 20 mg / kg daily (po) for 21 days. DETAILED DESCRIPTION

[0037] This specification is based on the inventors' data showing that the combination of an epidermal growth factor receptor inhibitor (such as osimertinib or cetuximab), a mitogen-activated protein kinase 1 / 2 inhibitor (such as cobimetinib, trametinib or TAK-733) and a cyclin-dependent kinase 4 / 6 inhibitor (such as palbociclib) provides a robust therapy for treating cancers with BRAF mutations or delaying their progression. In particular, this combination therapy does not require a BRAF inhibitor. Despite the fact that none of the compounds in the combination is an inhibitor of mutant BRAF, the combination therapy described herein has surprisingly demonstrated synergistic effects on cancers with BRAF mutations, as well as a strong efficacy of reducing tumor volume by up to about 95% in established animal models. The specification also provides compositions and kits that can be used to perform this combination therapy.

[0038] I. Definition

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which this disclosure belongs. All patents, applications, published applications, and other publications mentioned herein are incorporated by reference in their entireties. If a definition set forth in this section conflicts or is otherwise inconsistent with a definition set forth in a patent, application, or other publication incorporated herein by reference, the definition set forth in this section takes precedence over the definition incorporated herein by reference.

[0040] It will be appreciated that certain features of the present disclosure that are described for clarity in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, various features of the present disclosure that are described for brevity in the context of a single embodiment may also be provided separately or in any suitable subcombination. All combinations of embodiments relating to specific method steps, reagents, or conditions are expressly included in this disclosure and disclosed herein, just as if each and every combination were individually and expressly disclosed.

[0041] As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should also be noted that claims can be drafted to exclude any optional elements. As such, this statement is intended to serve as antecedent basis for use of exclusive terminology such as "solely," "only," and the like in connection with the recitation of claim elements, or for use of a "negative" limitation.

[0042] As used herein, the terms "including," "comprising," and "comprising" are used in their open, non-limiting sense. It should also be understood that the aspects and embodiments of the invention described herein may include "consisting of" and / or "consisting essentially of" the aspects and embodiments.

[0043] It should be understood that, regardless of whether the term "about" is explicitly used, each quantity given herein is intended to refer to the actual given value and also to the approximate value of this given value that is reasonably inferred based on ordinary skill in the art, including equivalent values ​​and approximate values ​​resulting from the experimental and / or measurement conditions of this given value.

[0044] As used herein, a subject who is "at risk" of developing a disease may or may not have had a detectable disease or disease symptom, and may or may not have shown a detectable disease or disease symptom, prior to the methods of treatment described herein. A subject who is "at risk" has one or more risk factors, which are measurable parameters associated with the development of a disease, such as cancer, as described herein and known in the art. A subject who is "at risk" may have one or more risk factors. A subject who has one or more risk factors has a higher probability of developing the disease than a subject who does not have the one or more risk factors.

[0045] As used herein, "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. More specific examples of such cancers include, but are not limited to, squamous cell carcinoma, lung cancer (including small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous carcinoma), peritoneal cancer, hepatocellular carcinoma, gastric or stomach cancer (including gastrointestinal cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney or renal cancer, liver cancer, prostate cancer, vulvar cancer, thyroid cancer, hepatic cancer, carcinoma) and various types of head and neck cancer, as well as B-cell lymphomas (including low-grade / follicular non-Hodgkin lymphoma (NHL); small lymphocytic (SL) NHL; intermediate-grade / follicular NHL; intermediate-grade diffuse NHL; high-grade immunoblastic NHL; high-grade lymphoblastic NHL; high-grade small non-cleaved cell NHL; giant disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's macroglobulinemia); chronic lymphocytic leukemia (CLL); acute lymphocytic leukemia (ALL); hairy cell leukemia; chronic myeloblastic leukemia; and post-transplant lymphoproliferative disorder (PTLD), as well as abnormal blood vessel proliferation associated with nevus hamartomatosis, edema (such as edema associated with brain tumors), and Meigs syndrome. Examples of cancer can include a primary tumor of any of the above types of cancer or a metastatic tumor at a second site derived from any of the above types of cancer. This definition includes both benign and malignant cancers as well as dormant tumors or micrometastases.

[0046] The terms "neoplastic cells," "tumor cells," or "cancer cells," used in the singular or plural, refer to cells that have undergone a malignant transformation that renders them pathological to the host organism. Primary cancer cells (i.e., cells obtained from the vicinity of the site of malignant transformation) can be readily distinguished from non-cancerous cells by established techniques, particularly histological examination. As used herein, the definition of cancer cells includes not only primary cancer cells, but also any cells derived from cancer progenitor cells. This includes metastatic cancer cells, as well as in vitro cultures and cell lines derived from cancer cells. When referring to cancer types that typically manifest as solid tumors, a "clinically detectable" tumor is one that is detectable based on the tumor mass; for example, it can be detected by procedures such as CAT scans, magnetic resonance imaging (MRI), X-rays, ultrasound, or palpation. Biochemical or immunological findings alone may not be sufficient to meet this definition.

[0047] As used herein, "carrier" includes pharmaceutically acceptable carriers, excipients, or stabilizers that are non-toxic to the cells or mammals to which they are exposed at the dosages and concentrations employed. Typically, a physiologically acceptable carrier is a pH-buffered aqueous solution. Non-limiting examples of physiologically acceptable carriers include buffers such as phosphates, citrates, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as TWEEN®. TM , polyethylene glycol (PEG) and PLURONICS TM .

[0048] As used herein, "delaying the progression of a disease" means postponing, hindering, slowing down, delaying, stabilizing and / or postponing the development of a disease (such as cancer). This delay can have different lengths of time, depending on the medical history and / or the individual being treated. It will be apparent to those skilled in the art that a sufficient or significant delay can actually encompass prevention, as the individual will not develop the disease. For example, the development of advanced cancers, such as metastases, may be delayed. A method of "delaying" the development of cancer is a method of reducing the probability of disease development and / or the extent of the disease within a given time frame when compared to not using the method. Such comparisons are typically based on clinical studies using a statistically significant number of subjects. Cancer development can be detectable using standard methods, including but not limited to computerized axial tomography (CAT scan), magnetic resonance imaging (MRI), ultrasound, coagulation tests, arteriography, biopsy, urine cytology and cystoscopy. Development can also refer to cancer progression, which may initially be undetectable and includes occurrence, recurrence and onset.

[0049] As used herein, the term "effective amount" or "therapeutically effective amount" of a substance is at least the minimum concentration required to achieve a measurable improvement or prevention of a particular disorder. The effective amount herein can vary according to a variety of factors, such as the patient's disease state, age, sex, and weight, and the ability of the substance to elicit a desired response in an individual. An effective amount is also an amount in which the therapeutically beneficial effects outweigh any toxic or deleterious effects of the treatment. With regard to cancer, an effective amount includes an amount sufficient to cause tumor shrinkage and / or reduce tumor growth rate (such as inhibiting tumor growth) or prevent or delay other unwanted cell proliferation in cancer. In some embodiments, an effective amount is an amount sufficient to delay the development of cancer. In some embodiments, an effective amount is an amount sufficient to prevent or delay recurrence. In some embodiments, an effective amount is an amount sufficient to reduce the recurrence rate of an individual. An effective amount can be given in one or more administrations. An effective amount of a drug or composition can: (i) reduce the number of cancer cells; (ii) reduce the size of a tumor; (iii) inhibit, delay, slow down, and preferably prevent cancer cells from infiltrating into peripheral organs to a certain extent; (iv) inhibit (i.e., slow down, and preferably prevent) tumor metastasis to a certain extent; (v) inhibit tumor growth; (vi) prevent or delay the occurrence and / or recurrence of a tumor; (vii) reduce the recurrence rate of a tumor, and / or (viii) alleviate one or more symptoms associated with cancer to a certain extent. An effective amount can be administered in one or more administrations. For the purposes of this disclosure, an effective amount of a drug, compound, or pharmaceutical composition is an amount sufficient to achieve a preventive or therapeutic treatment, either directly or indirectly. As understood in the clinical context, an effective amount of a drug, compound, or pharmaceutical composition may or may not be achieved in combination with another drug, compound, or pharmaceutical composition. Thus, an "effective amount" can be considered in the context of administering one or more therapeutic agents, and if the desired result can be achieved or is achieved in combination with one or more other agents, then administering a single agent in an effective amount can be considered.

[0050] As used herein, the term "inhibitor" or "antagonist" refers to a biological or chemical substance that interferes with or otherwise reduces the physiological and / or biochemical effects of another biological or chemical molecule. In some embodiments, an inhibitor or antagonist specifically binds to another molecule.

[0051] "Package insert" means instructions customarily included in commercial packages of pharmaceutical products, that contain information regarding the indications, usage, dosage, administration, contraindications, other drugs to be combined with the packaged product and / or warnings concerning the use of such drugs.

[0052] "Pharmaceutically acceptable salts" are salt forms that are non-toxic, biologically tolerable, or otherwise biologically suitable for administration to a subject. See generally Berge et al. (1977) J. Pharm. Sci. 66, 1. Specific pharmaceutically acceptable salts are those that are pharmacologically effective and suitable for contact with the tissues of a subject without excessive toxicity, irritation, or allergic reaction. Pharmaceutically acceptable salts include, but are not limited to, acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; or with organic acids such as acetic acid, oxalic acid, propionic acid, succinic acid, maleic acid, tartaric acid, etc. These salts can be derived from inorganic or organic acids. Non-limiting examples of pharmaceutically acceptable salts include, but are not limited to, sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, octanoates, acrylates, formates, isobutyrates, hexanoates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, Butyne-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, methylsulfonate, propylsulfonate, benzenesulfonate, xylenesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, gamma-hydroxybutyrate, glycolate, tartrate and mandelate. In some embodiments, when the acidic proton present in the parent compound is replaced by a metal ion (e.g., an alkali metal ion, an alkaline earth ion or an aluminum ion); or when coordinated with an organic base, a pharmaceutically acceptable salt is formed. Salts derived from pharmaceutically acceptable organic non-toxic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, tromethamine, tromethamine, dicyclohexylamine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-ethylglucamine, N-methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, amino acids such as lysine, arginine, histidine, etc. Examples of pharmaceutically acceptable base addition salts include those derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. In some embodiments, the organic non-toxic base is an L-amino acid such as L-lysine and L-arginine, tromethamine, N-ethylglucamine and N-methylglucamine. Acceptable inorganic bases include but are not limited to aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, etc.Lists of other suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pennsylvania, 1985.

[0053] A "solvate" is formed by the interaction of a solvent with a compound. Suitable solvents include, for example, water and alcohols (e.g., ethanol). Solvates include hydrates having any ratio of compound to water, such as monohydrates, dihydrates, and hemihydrates.

[0054] "Subject," "patient," or "individual" includes mammals, such as humans or other animals, and is typically a human. In some embodiments, the subject (e.g., patient) to whom the therapeutic agent and composition are administered is a mammal, typically a primate, such as a human. In some embodiments, the primate is a monkey or ape. The subject can be male or female and can be of any suitable age, including infants, young children, adolescents, adults, and elderly subjects. In some embodiments, the subject is a non-primate mammal, such as a rodent, dog, cat, farm animal (such as a cow or horse), etc.

[0055] As used herein, "tissue sample" or "cell sample" means a collection of similar cells obtained from a tissue of a subject or patient. The source of the tissue or cell sample can be solid tissue from a fresh, frozen and / or preserved organ or tissue sample or biopsy or aspirate; blood or any blood component; body fluids such as cerebrospinal fluid, amniotic fluid, peritoneal fluid or interstitial fluid; cells from any time of pregnancy or development of the subject. The tissue sample can also be a primary or cultured cell or cell line. Optionally, the tissue or cell sample is obtained from a diseased tissue / organ (such as cancer or tumor tissue). The tissue sample may contain compounds that are not naturally mixed with the tissue in nature, such as preservatives, anticoagulants, buffers, fixatives, nutrients, antibiotics, etc.

[0056] As used herein, the term "treatment" refers to a clinical intervention designed to have a beneficial and desired effect on the natural process of the treated individual or cell in the clinical pathology process. For the purposes of this disclosure, the desired effect of treatment includes but is not limited to reducing the disease progression rate, improving or alleviating the disease state and relaxing or improving prognosis. For example, if one or more symptoms associated with cancer are alleviated or eliminated, then successfully "treating" the individual, the alleviation or elimination includes but is not limited to reducing the proliferation (or destruction of cancer cells) of cancer cells, increasing cancer cell killing, reducing the symptoms caused by the disease, preventing disease spread, preventing disease recurrence, improving the quality of life of those patients with the disease, reducing the dosage of other drugs required for treating the disease, delaying the progress of the disease and / or prolonging the survival period of the individual.

[0057] II. Methods for treating and delaying progression of cancers harboring BRAF mutations

[0058] Provided herein are methods for treating or delaying the progression of cancer in a subject, comprising administering to the subject an effective amount of (a) an epidermal growth factor receptor (EGFR) inhibitor; (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor; and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor, wherein the subject has cancer or is at risk of developing cancer with a BRAF mutation.

[0059] In some embodiments, (a) is a small molecule or antibody (or antigen-binding fragment thereof) that specifically binds to EGFR or its ligand, and is optionally selected from one or more of cetuximab, panitumumab, zalutumumab, nimotuzumab, matuzumab, erlotinib, gefitinib, afatinib, lapatinib, osimertinib, brigatinib, and icotinib. In some embodiments, (b) is a small molecule or antibody (or antigen-binding fragment thereof) that specifically binds to MEK 1 / 2 or its ligand, and is optionally selected from one or more of trametinib, selumetinib, TAK-733, CI-1040, PD0325901, MEK162, AZD8330, GDC-0623, rameltinib, pimasetinib, RO4987655, RO5126766, WX-554, HL-085, and cobimetinib. See Chen et al. (2017) Molecules 22, 1551. In some embodiments, (c) is a small molecule or antibody (or antigen-binding fragment thereof) that specifically binds to CDK4 / 6 or its ligand, and is optionally selected from one or more of palbociclib, ribociclib, and abecinib. In some embodiments, the method does not include administering a BRAF inhibitor to the subject during administration of (a) an epidermal growth factor receptor (EGFR) inhibitor; (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor; and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor. In some embodiments, the method does not include administering an additional therapeutic agent during administration of (a) an epidermal growth factor receptor (EGFR) inhibitor; (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor; and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor. In some embodiments, the subject has received a BRAF inhibitor in a previous treatment cycle. In some embodiments, the subject has not received a BRAF inhibitor in a previous treatment cycle. In some embodiments, the subject is a human.

[0060] In one aspect, the methods disclosed herein can be used to treat cancers with BRAF mutations or to delay their progression. BRAF is a serine-threonine kinase, and BRAF mutations have been found in 28 primary cancers, including 6 primary melanomas, 12 short-term melanoma cultures, colorectal cancer, ovarian cancer, gliomas, lung cancer, breast cancer, sarcomas, etc. Without wishing to be bound by theory, it has been proposed that BRAF mutations lead to constitutive BRAF kinase activity, phosphorylation of MEK and ERK kinases, and sustained MAPK pathway signaling, thereby leading to tumor cell proliferation and survival. BRAF has been reported to mutate at several sites, but the vast majority of mutated BRAF is V600E, corresponding to a T to A transversion mutation at nucleotide 1796. BRAF mutations V600K, V600D, and D581V have also been observed. Detection of these mutations can be performed using conventional methods, such as the non-limiting examples reported in Lasota et al. (2015) Am. J. Surg. Pathol. 38, 1235. In some embodiments, a BRAF mutation is detected in a tissue or cell sample containing cancer cells from a subject. In some embodiments, the BRAF mutation is a somatic mutation. In some embodiments, the method is used to treat a cancer with a BRAF V600E mutation or to slow its progression. In some embodiments, the method is used to treat a cancer with a BRAF V600D mutation or to slow its progression. In some embodiments, the method is used to treat a cancer with a BRAF V600K mutation or to slow its progression. In some embodiments, the method is used to treat a cancer with a BRAF D581V mutation or to slow its progression. In some embodiments, the method does not include administering a BRAF inhibitor to the subject during administration of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the method does not include administering an additional therapeutic agent during administration of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the subject has received a BRAF inhibitor in a previous treatment cycle. In some embodiments, the subject has not received a BRAF inhibitor in a previous treatment cycle.

[0061] In some embodiments, the cancer is adenocarcinoma, squamous cell carcinoma, adenosquamous carcinoma, anaplastic carcinoma, large cell carcinoma, and small cell carcinoma. In some embodiments, the cancer is melanoma and carcinomas such as epithelial tumors, squamous cell tumors, basal cell tumors, transitional cell carcinomas, adenocarcinomas, adnexal or skin adnexal tumors, mucoepidermoid tumors, cystic, mucinous or serous tumors, ductal, lobular or medullary tumors, acinar cell tumors, and complex epithelial tumors. In some embodiments, the cancer is colon cancer, gastric cancer, lung cancer, breast cancer, pancreatic cancer, oral cancer, prostate cancer, reproductive system cancer, rectal cancer, liver cancer, kidney cancer, and ovarian cancer. In some embodiments, the cancer is advanced cancer, such as stage IV colorectal cancer. In some embodiments, the cancer is advanced colorectal cancer.

[0062] Also provided herein is a method for treating cancer or delaying its progression in a subject, comprising administering an effective amount of osimertinib or a salt thereof, cobimetinib or a salt thereof and palbociclib or a salt thereof to the subject, wherein the subject suffers from cancer or is at risk of developing a cancer with a BRAF mutation. On the other hand, also provided herein is a method for treating cancer or delaying its progression in a subject, comprising administering an effective amount of cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof to the subject, wherein the subject suffers from cancer or is at risk of developing a cancer with a BRAF mutation. On the other hand, also provided herein is a method for treating cancer or delaying its progression in a subject, comprising administering an effective amount of cetuximab, TAK-733 or a salt thereof and palbociclib or a salt thereof to the subject, wherein the subject suffers from cancer or is at risk of developing a cancer with a BRAF mutation. On the other hand, provided herein are methods for treating cancer or delaying its progression in a subject, comprising administering to the subject an effective amount of osimertinib or a salt thereof, TAK-733 or a salt thereof, and palbociclib or a salt thereof, wherein the subject suffers from cancer or is at risk of developing a cancer with a BRAF mutation. On the other hand, provided herein are methods for treating cancer or delaying its progression in a subject, comprising administering to the subject an effective amount of cetuximab, trametinib or a salt thereof, and palbociclib or a salt thereof, wherein the subject suffers from cancer or is at risk of developing a cancer with a BRAF mutation. On the other hand, provided herein are methods for treating cancer or delaying its progression in a subject, comprising administering to the subject an effective amount of osimertinib or a salt thereof, trametinib or a salt thereof, and palbociclib or a salt thereof, wherein the subject suffers from cancer or is at risk of developing a cancer with a BRAF mutation.

[0063] Osimertinib is an oral third-generation EGFR inhibitor that has been approved by the U.S. FDA and the European Commission (EC) for the treatment of non-small cell lung cancer with EGFR mutations. Osimertinib targets EGFR tyrosine kinase inhibitor (TKI)-sensitizing mutations, and in particular T790M, which often contributes to the acquisition of resistance to EGFR TKI therapy. Cobimetinib is a MEK inhibitor approved by the U.S. FDA for use in combination with the BRAF inhibitor vemurafenib for the treatment of metastatic melanoma with BRAF V600E or V600K mutations. Cobimetinib and vemurafenib target different components of the MAPK / ERK pathway: MEK and BRAF, respectively. Palbociclib is a CDK4 / 6 inhibitor approved by the U.S. FDA for the treatment of hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative advanced or metastatic breast cancer in combination with an aromatase inhibitor as endocrine-based therapy for postmenopausal women. Cetuximab is a chimeric monoclonal antibody administered by intravenous infusion and an EGFR inhibitor. It was approved by the US FDA in 2009 for the treatment of colon cancer with wild-type KRAS. TAK-733 is an orally bioavailable, non-ATP-competitive, small molecule MEK1 / 2 inhibitor that has completed Phase I clinical studies. Trametinib is a MEK inhibitor approved by the US FDA as a single agent or in combination with the BRAF inhibitor dabrafenib for the treatment of melanoma with BRAF V600E or V600K mutations. The structures of osimertinib, TAK-733, cobimetinib, trametinib, and palbociclib are shown below.

[0064]

[0065] In some embodiments, the method comprises administering a salt of osimertinib, cobimetinib, and palbociclib. In some embodiments, the salt is a pharmaceutically acceptable salt. Non-limiting examples of pharmaceutically acceptable salts include, but are not limited to, sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, octanoates, acrylates, formates, isobutyrates, hexanoates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butynes, benzoates, benzophenones, benzophenones, benzoates ... In some embodiments, the pharmaceutically acceptable salt is a fumarate salt. In some embodiments, the pharmaceutically acceptable salt is a methanesulfonate salt. In some embodiments, the pharmaceutically acceptable salt is a methanesulfonate salt. In some embodiments, the pharmaceutically acceptable salt is a fumarate salt. In some embodiments, the pharmaceutically acceptable salt is a methanesulfonate salt. In some embodiments, the method comprises administering a fumarate salt of osimertinib, cobimetinib, and palbociclib. In some embodiments, the method comprises administering a fumarate salt of osimertinib, cobimetinib, and palbociclib. In some embodiments, the method comprises administering a fumarate salt of osimertinib, cobimetinib, and palbociclib. In some embodiments, the method comprises administering a fumarate salt of osimertinib, cobimetinib, and palbociclib. In some embodiments, the method comprises administering a fumarate salt of osimertinib and cobimetinib and palbociclib. In some embodiments, the method comprises administering the mesylate salt of osimertinib and the fumarate salts of cobimetinib and palbociclib. In some embodiments, the method comprises administering osimertinib, cobimetinib, and palbociclib, or a solvate or salt of any one of the foregoing.

[0066] The method can comprise administering any of the compositions or kits described herein.

[0067] On the other hand, the method comprises administering an effective amount of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the daily dose of osimertinib or a salt thereof is about 0.25-0.5 mg / kg, 0.5-1 mg / kg, about 1-1.5 mg / kg, about 1.5-2 mg / kg, about 2-2.5 mg / kg, about 2.5-3 mg / kg, about 3-3.5 mg / kg, about 3.5-4 mg / kg, or about 0.5-3 mg / kg of osimertinib. In some embodiments, the daily dose of osimertinib or a salt thereof is about 20 mg, about 40 mg, about 80 mg, or about 160 mg of osimertinib. In some embodiments, the daily dose of osimertinib or a salt thereof is less than about 20 mg, about 40 mg, about 80 mg, or about 160 mg of osimertinib. In some embodiments, the dose of osimertinib or a salt thereof is an amount of about 20-240 mg, about 20-40 mg, about 40-80 mg, about 80-160 mg, about 160-240 mg, or about 40-160 mg of osimertinib. In some embodiments, the daily dose of osimertinib or a salt thereof is an amount of about 0.6-2.7 mg / kg of osimertinib. In some embodiments, osimertinib is in the form of a mesylate salt. The amount of the inhibitor described herein and throughout the specification refers to the amount of the inhibitor without taking into account the weight of the counterion if the inhibitor is present in salt form. For example, the term "80 mg of osimertinib" may include, but is not limited to, 80 mg of osimertinib in a salt-free form or 95.4 mg of osimertinib mesylate.

[0068] In some embodiments, the daily dose of cobimetinib or its salt is about 0.1-0.25mg / kg, about 0.25-0.5mg / kg, about 0.5-0.75mg / kg, about 0.75-1mg / kg, about 1-1.25mg / kg, about 1.25-1.5mg / kg, about 1.5-1.75mg / kg, about 1.75-2mg / kg or about 0.25-1mg / kg cobimetinib. In some embodiments, the daily dose of cobimetinib or its salt is about 10mg, about 20mg, about 40mg or about 60mg cobimetinib. In some embodiments, the daily dose of cobimetinib or its salt is less than about 10mg, about 20mg, about 40mg or about 60mg cobimetinib. In some embodiments, the daily dose of cobimetinib or its salt is about 3.5-100mg, about 3.5-10mg, about 10-20mg, about 20-40mg, about 40-60mg, about 20-60mg or about 60-100mg of cobimetinib. In some embodiments, the daily dose of cobimetinib or its salt is about 0.3-1mg / kg of cobimetinib. In some embodiments, cobimetinib is in the form of its hemifumarate salt.

[0069] In some embodiments, the daily dose of palbociclib or its salt is an amount of about 0.5-1 mg / kg, about 1.5-2 mg / kg, about 2-2.5 mg / kg, about 2.5-3 mg / kg, about 3-3.5 mg / kg, about 1-2.5 mg / kg, about 1-3 mg / kg, or about 3-5 mg / kg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is an amount of about 50 mg, about 75 mg, about 100 mg, or about 125 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is an amount of less than about 50 mg, about 75 mg, about 100 mg, or about 125 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is about 25-225 mg, about 25-50 mg, about 50-75 mg, about 75-125 mg, about 125-150 mg, about 150-200 mg, or about 200-225 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is about 1.25-2.1 mg / kg of palbociclib. In some embodiments, the method comprises administering osimertinib or its solvate or salt, cobimetinib or its solvate or salt, and palbociclib or its solvate or salt.

[0070] In some embodiments, the daily dose of osimertinib or a salt thereof is an amount that is greater than about 20%, or greater than about 25%, or greater than about 30%, or greater than about 35%, or greater than about 40%, or greater than about 45%, or greater than about 50%, or greater than about 55%, or greater than about 60%, or greater than about 65%, or greater than about 70%, or greater than about 75%, or greater than about 80%, or greater than about 85%, or greater than about 90%, or greater than 95% by weight of the combined daily dose of osimertinib, cobimetinib, and palbociclib, or salts of the foregoing. In some embodiments, the daily dose of osimertinib or a salt thereof is an amount that is less than about 20%, or less than about 25%, or less than about 30%, or less than about 35%, or less than about 40%, or less than about 45%, or less than about 50%, or less than about 55%, or less than about 60%, or less than about 65%, or less than about 70%, or less than about 75%, or less than about 80%, or less than about 85%, or less than about 90%, or less than 95% by weight of the combined daily dose of osimertinib, cobimetinib, and palbociclib, or salts of the foregoing.

[0071] In some embodiments, the daily dose of cobimetinib or a salt thereof is an amount that is greater than about 20%, or greater than about 25%, or greater than about 30%, or greater than about 35%, or greater than about 40%, or greater than about 45%, or greater than about 50%, or greater than about 55%, or greater than about 60%, or greater than about 65%, or greater than about 70%, or greater than about 75%, or greater than about 80%, or greater than about 85%, or greater than about 90%, or greater than 95% by weight of the combined daily dose of osimertinib, cobimetinib, and palbociclib, or salts of the foregoing. In some embodiments, the daily dose of cobimetinib or a salt thereof is an amount that is less than about 20%, or less than about 25%, or less than about 30%, or less than about 35%, or less than about 40%, or less than about 45%, or less than about 50%, or less than about 55%, or less than about 60%, or less than about 65%, or less than about 70%, or less than about 75%, or less than about 80%, or less than about 85%, or less than about 90%, or less than 95% by weight of the combined daily dose of osimertinib, cobimetinib, and palbociclib, or salts of the foregoing.

[0072] In some embodiments, the daily dose of palbociclib or a salt thereof is an amount that is greater than about 20%, or greater than about 25%, or greater than about 30%, or greater than about 35%, or greater than about 40%, or greater than about 45%, or greater than about 50%, or greater than about 55%, or greater than about 60%, or greater than about 65%, or greater than about 70%, or greater than about 75%, or greater than about 80%, or greater than about 85%, or greater than about 90%, or greater than 95% by weight of the combined daily dose of osimertinib, cobimetinib, and palbociclib, or a salt of the foregoing. In some embodiments, the daily dose of palbociclib or a salt thereof is an amount that is less than about 20%, or less than about 25%, or less than about 30%, or less than about 35%, or less than about 40%, or less than about 45%, or less than about 50%, or less than about 55%, or less than about 60%, or less than about 65%, or less than about 70%, or less than about 75%, or less than about 80%, or less than about 85%, or less than about 90%, or less than 95% by weight of the combined daily dose of osimertinib, cobimetinib, and palbociclib, or a salt of the foregoing.

[0073] In another aspect, the method comprises administering an effective amount of cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof. In some embodiments, the dose of cetuximab is about 0.1-20 mg / kg, about 0.1-0.5 mg / kg, about 0.5-1 mg / kg, about 1-2 mg / kg, about 2-3 mg / kg, about 3-5 mg / kg, about 5-7.5 mg / kg, about 7.5-10 mg / kg, about 10-15 mg / kg, about 15-20 mg / kg or about 0.1-10 mg / kg. In some embodiments, the dose of cetuximab is about 150-200 mg / kg. 2, about 200-250mg / m 2 , about 250-300mg / m 2 , about 300-400mg / m 2 , about 400-500mg / m 2 , about 500-750mg / m 2 , about 150-250mg / m 2 , about 250-400mg / m 2 or about 400-750 mg / m 2 In some embodiments, cetuximab is infused over 30-180 minutes, about 30-60 minutes, about 60-120 minutes, or about 120-180 minutes. In some embodiments, the maximum infusion rate is about 5 mL / min or about 10 mL / min. In some embodiments, cetuximab is administered about daily, about weekly, about every two weeks, about every three weeks, or about every four weeks. In some embodiments, the dose of cetuximab is about 500 mg / m2 infused over 60-120 minutes every two weeks. 2 In some embodiments, cetuximab is administered according to a schedule that includes an initial dose followed by several subsequent doses. In some embodiments, the initial dose is about 250-500 mg / m 2 , about 250-300mg / m 2 , about 300-400 mg / m or about 400-500 mg / m 2 In some embodiments, the initial dose is about 250 mg / m 2 , about 400mg / m 2 or about 500 mg / m 2 In some embodiments, subsequent doses are about 50-300 mg / m 2 , about 50-150mg / m 2 , about 150-200mg / m 2 or about 200-300 mg / m 2 In some embodiments, the subsequent dose is about 50 mg / m 2 , about 150mg / m 2 or about 250 mg / m 2 In some embodiments, 400 mg / m2 is infused over 120 minutes. 2 , followed by weekly infusions of 250 mg / m2 over 60 minutes 2 In some embodiments, cetuximab is administered as a 400 mg / m2 infusion over 120 minutes. 2 , followed by weekly infusions of 150 mg / m2 over 60 minutes 2 Cetuximab was administered in a controlled manner.

[0074] In some embodiments, the daily dose of cobimetinib or its salt is about 0.1-0.25mg / kg, about 0.25-0.5mg / kg, about 0.5-0.75mg / kg, about 0.75-1mg / kg, about 1-1.25mg / kg, about 1.25-1.5mg / kg, about 1.5-1.75mg / kg, about 1.75-2mg / kg or about 0.25-1mg / kg cobimetinib. In some embodiments, the daily dose of cobimetinib or its salt is about 10mg, about 20mg, about 40mg or about 60mg cobimetinib. In some embodiments, the daily dose of cobimetinib or its salt is less than about 10mg, about 20mg, about 40mg or about 60mg cobimetinib. In some embodiments, the daily dose of cobimetinib or its salt is about 3.5-100mg, about 3.5-10mg, about 10-20mg, about 20-40mg, about 40-60mg, about 20-60mg or about 60-100mg of cobimetinib. In some embodiments, the daily dose of cobimetinib or its salt is about 0.3-1mg / kg of cobimetinib. In some embodiments, cobimetinib is in the form of its hemifumarate salt.

[0075] In some embodiments, the daily dose of palbociclib or its salt is an amount of about 0.5-1 mg / kg, about 1.5-2 mg / kg, about 2-2.5 mg / kg, about 2.5-3 mg / kg, about 3-3.5 mg / kg, about 1-2.5 mg / kg, about 1-3 mg / kg, or about 3-5 mg / kg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is an amount of about 50 mg, about 75 mg, about 100 mg, or about 125 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is an amount of less than about 50 mg, about 75 mg, about 100 mg, or about 125 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is about 25-225 mg, about 25-50 mg, about 50-75 mg, about 75-125 mg, about 125-150 mg, about 150-200 mg, or about 200-225 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is about 1.25-2.1 mg / kg of palbociclib. In some embodiments, the method comprises administering cetuximab, cobimetinib, or a solvate or salt thereof, and palbociclib, or a solvate or salt thereof.

[0076] In another aspect, the method comprises administering effective amounts of cetuximab, TAK-733 or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the daily dose of TAK-733 or a salt thereof is an amount of about 0.001-1 mg / kg, about 0.001-0.002 mg / kg, about 0.002-0.005 mg / kg, about 0.005-0.01 mg / kg, about 0.01-0.05 mg / kg, about 0.05-0.1 mg / kg, about 0.1-0.2 mg / kg, about 0.2-0.3 mg / kg, about 0.3-0.4 mg / kg, about 0.4-0.5 mg / kg, about 0.5-0.6 mg / kg, about 0.6-0.7 mg / kg, about 0.7-0.8 mg / kg, about 0.8-0.9 mg / kg, or about 0.9-1 mg / kg of TAK-733. In some embodiments, the daily dose of TAK-733 or a salt thereof is an amount of about 10 mg, about 15 mg, about 20 mg, or about 25 mg of TAK-733. In some embodiments, the daily dose of TAK-733 or a salt thereof is an amount of less than about 10 mg, about 15 mg, about 20 mg, or about 25 mg of TAK-733. In some embodiments, the daily dose of TAK-733 or a salt thereof is an amount of about 0.1-25 mg, about 0.1-1 mg, about 1-5 mg, about 5-10 mg, about 8-16 mg, about 10-15 mg, about 15-20 mg, or about 20-25 mg of TAK-733.

[0077] In some embodiments, the dose of cetuximab is about 0.1-20 mg / kg, about 0.1-0.5 mg / kg, about 0.5-1 mg / kg, about 1-2 mg / kg, about 2-3 mg / kg, about 3-5 mg / kg, about 5-7.5 mg / kg, about 7.5-10 mg / kg, about 10-15 mg / kg, about 15-20 mg / kg, or about 0.1-10 mg / kg. In some embodiments, the dose of cetuximab is about 150-200 mg / kg. 2 , about 200-250mg / m 2 , about 250-300mg / m 2 , about 300-400mg / m 2 , about 400-500mg / m 2 , about 500-750mg / m 2 , about 150-250mg / m 2 , about 250-400mg / m 2 or about 400-750 mg / m 2In some embodiments, cetuximab is infused over 30-180 minutes, about 30-60 minutes, about 60-120 minutes, or about 120-180 minutes. In some embodiments, the maximum infusion rate is about 5 mL / min or about 10 mL / min. In some embodiments, cetuximab is administered about daily, about weekly, about every two weeks, about every three weeks, or about every four weeks. In some embodiments, the dose of cetuximab is about 500 mg / m2 infused over 60-120 minutes every two weeks. 2 In some embodiments, cetuximab is administered according to a schedule that includes an initial dose followed by several subsequent doses. In some embodiments, the initial dose is about 250-500 mg / m 2 , about 250-300mg / m 2 , about 300-400 mg / m or about 400-500 mg / m 2 In some embodiments, the initial dose is about 250 mg / m 2 , about 400mg / m 2 or about 500 mg / m 2 In some embodiments, subsequent doses are about 50-300 mg / m 2 , about 50-150mg / m 2 , about 150-200mg / m 2 or about 200-300 mg / m 2 In some embodiments, the subsequent dose is about 50 mg / m 2 , about 150mg / m 2 or about 250 mg / m 2 In some embodiments, 400 mg / m2 is infused over 120 minutes. 2 , followed by weekly infusions of 250 mg / m2 over 60 minutes 2 In some embodiments, cetuximab is administered as a 400 mg / m2 infusion over 120 minutes. 2 , followed by weekly infusions of 150 mg / m2 over 60 minutes 2 Cetuximab was administered in a controlled manner.

[0078] In some embodiments, the daily dose of palbociclib or its salt is an amount of about 0.5-1 mg / kg, about 1.5-2 mg / kg, about 2-2.5 mg / kg, about 2.5-3 mg / kg, about 3-3.5 mg / kg, about 1-2.5 mg / kg, about 1-3 mg / kg, or about 3-5 mg / kg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is an amount of about 50 mg, about 75 mg, about 100 mg, or about 125 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is an amount of less than about 50 mg, about 75 mg, about 100 mg, or about 125 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is about 25-225 mg, about 25-50 mg, about 50-75 mg, about 75-125 mg, about 125-150 mg, about 150-200 mg, or about 200-225 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is about 1.25-2.1 mg / kg of palbociclib. In some embodiments, the method comprises administering cetuximab, TAK-733, or a solvate or salt thereof, and palbociclib, or a solvate or salt thereof.

[0079] On the other hand, the method comprises administering an effective amount of osimertinib or a salt thereof, TAK-733 or a salt thereof and palbociclib or a salt thereof. In some embodiments, the daily dose of osimertinib or a salt thereof is about 0.25-0.5 mg / kg, 0.5-1 mg / kg, about 1-1.5 mg / kg, about 1.5-2 mg / kg, about 2-2.5 mg / kg, about 2.5-3 mg / kg, about 3-3.5 mg / kg, about 3.5-4 mg / kg or about 0.5-3 mg / kg of osimertinib. In some embodiments, the daily dose of osimertinib or a salt thereof is about 20 mg, about 40 mg, about 80 mg or about 160 mg of osimertinib. In some embodiments, the daily dose of osimertinib or a salt thereof is less than about 20 mg, about 40 mg, about 80 mg or about 160 mg of osimertinib. In some embodiments, the dosage of osimertinib or a salt thereof is about 20-240 mg, about 20-40 mg, about 40-80 mg, about 80-160 mg, about 160-240 mg, or about 40-160 mg of osimertinib. In some embodiments, the daily dose of osimertinib or a salt thereof is about 0.6-2.7 mg / kg of osimertinib. In some embodiments, osimertinib is in the form of a mesylate salt.

[0080] In some embodiments, the daily dose of TAK-733 or a salt thereof is an amount of about 0.001-1 mg / kg, about 0.001-0.002 mg / kg, about 0.002-0.005 mg / kg, about 0.005-0.01 mg / kg, about 0.01-0.05 mg / kg, about 0.05-0.1 mg / kg, about 0.1-0.2 mg / kg, about 0.2-0.3 mg / kg, about 0.3-0.4 mg / kg, about 0.4-0.5 mg / kg, about 0.5-0.6 mg / kg, about 0.6-0.7 mg / kg, about 0.7-0.8 mg / kg, about 0.8-0.9 mg / kg, or about 0.9-1 mg / kg of TAK-733. In some embodiments, the daily dose of TAK-733 or a salt thereof is an amount of about 10 mg, about 15 mg, about 20 mg, or about 25 mg of TAK-733. In some embodiments, the daily dose of TAK-733 or a salt thereof is an amount of less than about 10 mg, about 15 mg, about 20 mg, or about 25 mg of TAK-733. In some embodiments, the daily dose of TAK-733 or a salt thereof is an amount of about 0.1-25 mg, about 0.1-1 mg, about 1-5 mg, about 5-10 mg, about 8-16 mg, about 10-15 mg, about 15-20 mg, or about 20-25 mg of TAK-733.

[0081] In some embodiments, the daily dose of palbociclib or its salt is an amount of about 0.5-1 mg / kg, about 1.5-2 mg / kg, about 2-2.5 mg / kg, about 2.5-3 mg / kg, about 3-3.5 mg / kg, about 1-2.5 mg / kg, about 1-3 mg / kg, or about 3-5 mg / kg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is an amount of about 50 mg, about 75 mg, about 100 mg, or about 125 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is an amount of less than about 50 mg, about 75 mg, about 100 mg, or about 125 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is about 25-225 mg, about 25-50 mg, about 50-75 mg, about 75-125 mg, about 125-150 mg, about 150-200 mg, or about 200-225 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is about 1.25-2.1 mg / kg of palbociclib. In some embodiments, the method comprises administering osimertinib or its solvate or salt, TAK-733 or its solvate or salt, and palbociclib or its solvate or salt.

[0082] In another aspect, the method comprises administering an effective amount of cetuximab, trametinib or a salt thereof and palbociclib or a salt thereof. In some embodiments, the daily dose of trametinib or a salt thereof is about 0.01-1 mg / kg, about 0.01-0.02 mg / kg, about 0.02-0.03 mg / kg, about 0.03-0.05 mg / kg, about 0.05-0.08 mg / kg, about 0.08-0.1 mg / kg, about 0.1-0.2 mg / kg, about 0.2-0.3 mg / kg, about 0.3-0.4 mg / kg, about 0.4-0.5 mg / kg, about 0.5-0.6 mg / kg, about 0.6-0.7 mg / kg, about 0.7-0.8 mg / kg, about 0.8-0.9 mg / kg, or about 0.9-1 mg / kg of trametinib. In some embodiments, the daily dose of trametinib or a salt thereof is an amount of about 0.5 mg, about 1 mg, about 2 mg, or about 4 mg trametinib. In some embodiments, the daily dose of trametinib or a salt thereof is an amount of less than about 0.5 mg, about 1 mg, about 2 mg, or about 4 mg trametinib. In some embodiments, the daily dose of trametinib or a salt thereof is an amount of about 0.1-25 mg, about 0.1-0.5 mg, about 0.5-1 mg, about 1-2 mg, about 2-4 mg, about 4-10 mg, or about 0.5-2 mg trametinib.

[0083] In some embodiments, the dose of cetuximab is about 0.1-20 mg / kg, about 0.1-0.5 mg / kg, about 0.5-1 mg / kg, about 1-2 mg / kg, about 2-3 mg / kg, about 3-5 mg / kg, about 5-7.5 mg / kg, about 7.5-10 mg / kg, about 10-15 mg / kg, about 15-20 mg / kg, or about 0.1-10 mg / kg. In some embodiments, the dose of cetuximab is about 150-200 mg / kg. 2 , about 200-250mg / m 2 , about 250-300mg / m 2 , about 300-400mg / m 2 , about 400-500mg / m 2 , about 500-750mg / m 2 , about 150-250mg / m 2 , about 250-400mg / m 2 or about 400-750 mg / m 2In some embodiments, cetuximab is infused over 30-180 minutes, about 30-60 minutes, about 60-120 minutes, or about 120-180 minutes. In some embodiments, the maximum infusion rate is about 5 mL / min or about 10 mL / min. In some embodiments, cetuximab is administered about daily, about weekly, about every two weeks, about every three weeks, or about every four weeks. In some embodiments, the dose of cetuximab is about 500 mg / m2 infused over 60-120 minutes every two weeks. 2 In some embodiments, cetuximab is administered according to a schedule that includes an initial dose followed by several subsequent doses. In some embodiments, the initial dose is about 250-500 mg / m 2 , about 250-300mg / m 2 , about 300-400 mg / m or about 400-500 mg / m 2 In some embodiments, the initial dose is about 250 mg / m 2 , about 400mg / m 2 or about 500 mg / m 2 In some embodiments, subsequent doses are about 50-300 mg / m 2 , about 50-150mg / m 2 , about 150-200mg / m 2 or about 200-300 mg / m 2 In some embodiments, the subsequent dose is about 50 mg / m 2 , about 150mg / m 2 or about 250 mg / m 2 In some embodiments, 400 mg / m2 is infused over 120 minutes. 2 , followed by weekly infusions of 250 mg / m2 over 60 minutes 2 In some embodiments, cetuximab is administered as a 400 mg / m2 infusion over 120 minutes. 2 , followed by weekly infusions of 150 mg / m2 over 60 minutes 2 Cetuximab was administered in a controlled manner.

[0084] In some embodiments, the daily dose of palbociclib or its salt is an amount of about 0.5-1 mg / kg, about 1.5-2 mg / kg, about 2-2.5 mg / kg, about 2.5-3 mg / kg, about 3-3.5 mg / kg, about 1-2.5 mg / kg, about 1-3 mg / kg, or about 3-5 mg / kg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is an amount of about 50 mg, about 75 mg, about 100 mg, or about 125 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is an amount of less than about 50 mg, about 75 mg, about 100 mg, or about 125 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is about 25-225 mg, about 25-50 mg, about 50-75 mg, about 75-125 mg, about 125-150 mg, about 150-200 mg, or about 200-225 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is about 1.25-2.1 mg / kg of palbociclib. In some embodiments, the method comprises administering cetuximab, TAK-733, or a solvate or salt thereof, and palbociclib, or a solvate or salt thereof.

[0085] In another aspect, the method comprises administering an effective amount of osimertinib or a salt thereof, trametinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the daily dose of osimertinib or a salt thereof is about 0.25-0.5 mg / kg, 0.5-1 mg / kg, about 1-1.5 mg / kg, about 1.5-2 mg / kg, about 2-2.5 mg / kg, about 2.5-3 mg / kg, about 3-3.5 mg / kg, about 3.5-4 mg / kg, or about 0.5-3 mg / kg of osimertinib. In some embodiments, the daily dose of osimertinib or a salt thereof is about 20 mg, about 40 mg, about 80 mg, or about 160 mg of osimertinib. In some embodiments, the daily dose of osimertinib or a salt thereof is less than about 20 mg, about 40 mg, about 80 mg, or about 160 mg of osimertinib. In some embodiments, the dosage of osimertinib or a salt thereof is about 20-240 mg, about 20-40 mg, about 40-80 mg, about 80-160 mg, about 160-240 mg, or about 40-160 mg of osimertinib. In some embodiments, the daily dose of osimertinib or a salt thereof is about 0.6-2.7 mg / kg of osimertinib. In some embodiments, osimertinib is in the form of a mesylate salt.

[0086] In some embodiments, the daily dose of trametinib or a salt thereof is about 0.01-1 mg / kg, about 0.01-0.02 mg / kg, about 0.02-0.03 mg / kg, about 0.03-0.05 mg / kg, about 0.05-0.08 mg / kg, about 0.08-0.1 mg / kg, about 0.1-0.2 mg / kg, about 0.2-0.3 mg / kg, about 0.3-0.4 mg / kg, about 0.4-0.5 mg / kg, about 0.5-0.6 mg / kg, about 0.6-0.7 mg / kg, about 0.7-0.8 mg / kg, about 0.8-0.9 mg / kg, or about 0.9-1 mg / kg trametinib. In some embodiments, the daily dose of trametinib or a salt thereof is about 0.5 mg, about 1 mg, about 2 mg, or about 4 mg trametinib. In some embodiments, the daily dose of trametinib or a salt thereof is an amount of less than about 0.5 mg, about 1 mg, about 2 mg, or about 4 mg of trametinib. In some embodiments, the daily dose of trametinib or a salt thereof is an amount of about 0.1-25 mg, about 0.1-0.5 mg, about 0.5-1 mg, about 1-2 mg, about 2-4 mg, about 4-10 mg, or about 0.5-2 mg of trametinib.

[0087] In some embodiments, the daily dose of palbociclib or its salt is an amount of about 0.5-1 mg / kg, about 1.5-2 mg / kg, about 2-2.5 mg / kg, about 2.5-3 mg / kg, about 3-3.5 mg / kg, about 1-2.5 mg / kg, about 1-3 mg / kg, or about 3-5 mg / kg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is an amount of about 50 mg, about 75 mg, about 100 mg, or about 125 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is an amount of less than about 50 mg, about 75 mg, about 100 mg, or about 125 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is about 25-225 mg, about 25-50 mg, about 50-75 mg, about 75-125 mg, about 125-150 mg, about 150-200 mg, or about 200-225 mg of palbociclib. In some embodiments, the daily dose of palbociclib or its salt is about 1.25-2.1 mg / kg of palbociclib. In some embodiments, the method comprises administering osimertinib or its solvate or salt, trametinib or its solvate or salt, and palbociclib or its solvate or salt.

[0088] On the other hand, the method provides for administering an effective amount of an EGFR inhibitor (e.g., osimertinib or cetuximab), a MEK 1 / 2 inhibitor (e.g., cobimetinib, trametinib or TAK-733) and a CDK 4 / 6 inhibitor (e.g., palbociclib) until disease progression or unacceptable toxicity occurs. In some embodiments, the method provides for administering an effective amount of an EGFR inhibitor (e.g., osimertinib or cetuximab), a MEK 1 / 2 inhibitor (e.g., cobimetinib, trametinib or TAK-733) and a CDK 4 / 6 inhibitor (e.g., palbociclib) for at least about 1-2 weeks, about 2-3 weeks, about 3-4 weeks, about 4-5 weeks, about 5-6 weeks, about 6-7 weeks, about 7-8 weeks, about 8-9 weeks, about 9-10 weeks, about 2-3 months, about 3-4 months, about 4-5 months, about 5-6 months, about 6-12 months or about 12-24 months. In some embodiments, an effective amount of an EGFR inhibitor (e.g., osimertinib or cetuximab), a MEK 1 / 2 inhibitor (e.g., cobimetinib, trametinib, or TAK-733), and a CDK 4 / 6 inhibitor (e.g., palbociclib) is administered for 21 consecutive days, followed by a 7-day rest period to constitute a complete cycle of 28 days. In some embodiments, an effective amount of an EGFR inhibitor (e.g., osimertinib or cetuximab), a MEK 1 / 2 inhibitor (e.g., cobimetinib, trametinib, or TAK-733), and a CDK 4 / 6 inhibitor (e.g., palbociclib) is administered to a subject for one, two, three, four, five, six, seven, eight, nine, ten, or more 28-day cycles. In some embodiments, the EGFR inhibitor (e.g., osimertinib or cetuximab), MEK 1 / 2 inhibitor (e.g., cobimetinib, trametinib, or TAK-733), and CDK 4 / 6 inhibitor (e.g., palbociclib) are administered for different durations.

[0089] On the other hand, an effective amount of an EGFR inhibitor (e.g., osimertinib or cetuximab), a MEK 1 / 2 inhibitor (e.g., cobimetinib, trametinib or TAK-733) and a CDK 4 / 6 inhibitor (e.g., palbociclib) are formulated into a composition. On the other hand, an effective amount of an EGFR inhibitor (e.g., osimertinib or cetuximab), a MEK 1 / 2 inhibitor (e.g., cobimetinib, trametinib or TAK-733) and a CDK 4 / 6 inhibitor (e.g., palbociclib) are formulated separately. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof and palbociclib or a salt thereof are formulated into a composition or separate composition for oral administration. In some embodiments, osimertinib or a salt thereof, TAK-733 or a salt thereof and palbociclib or a salt thereof are formulated into a composition or separate composition for oral administration. In some embodiments, osimertinib or a salt thereof, trametinib or a salt thereof, and palbociclib or a salt thereof are formulated into a composition or separate compositions for oral administration. In some embodiments, cobimetinib or a salt thereof and palbociclib or a salt thereof are formulated into a composition or separate compositions for oral administration. In some embodiments, TAK-733 or a salt thereof and palbociclib or a salt thereof are formulated into a composition or separate compositions for oral administration. In some embodiments, trametinib or a salt thereof and palbociclib or a salt thereof are formulated into a composition or separate compositions for oral administration. For oral administration, the method may include formulating the compound into a solid form, such as a tablet or capsule, or into a solution, emulsion, or suspension. In order to prepare an oral composition, the compound may be formulated to obtain a daily dose as described herein. In some embodiments, cetuximab is formulated for intravenous infusion.

[0090] In some embodiments, EGFR inhibitors (e.g., osimertinib or cetuximab), MEK 1 / 2 inhibitors (e.g., cobimetinib, trametinib or TAK-733) and CDK 4 / 6 inhibitors (e.g., palbociclib) are formulated into the same form. In some embodiments, osimertinib or a salt thereof, cobimetinib or a salt thereof and / or palbociclib or a salt thereof are formulated into a solid form, such as a tablet or capsule. In some embodiments, osimertinib or a salt thereof, TAK-733 or a salt thereof and / or palbociclib or a salt thereof are formulated into a solid form, such as a tablet or capsule. In some embodiments, osimertinib or a salt thereof, trametinib or a salt thereof and / or palbociclib or a salt thereof are formulated into a solid form, such as a tablet or capsule. In some embodiments, cetuximab, cobimetinib or a salt thereof and / or palbociclib or a salt thereof are formulated into a liquid form, such as a suspension, solution, emulsion or syrup, or can be lyophilized. In some embodiments, cetuximab, TAK-733 or a salt thereof and / or palbociclib or a salt thereof are formulated into a liquid form, such as a suspension, solution, emulsion or syrup, or can be lyophilized. In some embodiments, cetuximab, trametinib or a salt thereof and / or palbociclib or a salt thereof are formulated into a liquid form, such as a suspension, solution, emulsion or syrup, or can be lyophilized.

[0091] On the other hand, EGFR inhibitors (e.g., osimertinib or cetuximab), MEK 1 / 2 inhibitors (e.g., cobimetinib, trametinib or TAK-733) and CDK 4 / 6 inhibitors (e.g., palbociclib) are administered simultaneously or intermittently. In some embodiments, EGFR inhibitors (e.g., osimertinib or cetuximab), MEK 1 / 2 inhibitors (e.g., cobimetinib, trametinib or TAK-733) and CDK 4 / 6 inhibitors (e.g., palbociclib) are formulated into a composition and administered as a composition. In some embodiments, EGFR inhibitors (e.g., osimertinib or cetuximab), MEK 1 / 2 inhibitors (e.g., cobimetinib, trametinib or TAK-733) and CDK 4 / 6 inhibitors (e.g., palbociclib) are formulated separately and administered simultaneously. In some embodiments, the EGFR inhibitor (e.g., osimertinib or cetuximab), MEK 1 / 2 inhibitor (e.g., cobimetinib, trametinib, or TAK-733), and CDK 4 / 6 inhibitor (e.g., palbociclib) are formulated separately and administered intermittently. In some embodiments, the EGFR inhibitor (e.g., osimertinib or cetuximab), MEK 1 / 2 inhibitor (e.g., cobimetinib, trametinib, or TAK-733), and CDK 4 / 6 inhibitor (e.g., palbociclib) are formulated separately and administered with different dosing frequencies.

[0092] In another aspect, the method comprises administering an effective amount of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, osimertinib or a salt thereof is administered before cobimetinib or a salt thereof and palbociclib or a salt thereof. In some embodiments, cobimetinib or a salt thereof is administered before osimertinib or a salt thereof and palbociclib or a salt thereof. In some embodiments, palbociclib or a salt thereof is administered before osimertinib or a salt thereof and cobimetinib or a salt thereof. In some embodiments, osimertinib or a salt thereof and cobimetinib or a salt thereof are administered together in the same or separate compositions. In some embodiments, osimertinib or a salt thereof and palbociclib or a salt thereof are administered together in the same or separate compositions. In some embodiments, cobimetinib or a salt thereof and palbociclib or a salt thereof are administered together in the same or separate compositions.

[0093] On the other hand, the method comprises administering an effective amount of cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof. In some embodiments, cetuximab is administered before cobimetinib or a salt thereof and palbociclib or a salt thereof. In some embodiments, cobimetinib or a salt thereof is administered before cetuximab and palbociclib or a salt thereof. In some embodiments, palbociclib or a salt thereof is administered before cetuximab and cobimetinib or a salt thereof. In some embodiments, cetuximab and cobimetinib or a salt thereof are administered together in the same or separate compositions. In some embodiments, cetuximab and palbociclib or a salt thereof are administered together in the same or separate compositions. In some embodiments, cobimetinib or a salt thereof and palbociclib or a salt thereof are administered together in the same or separate compositions. In some embodiments, cetuximab is administered as a composition separate from cobimetinib or a salt thereof and palbociclib or a salt thereof. In some embodiments, cetuximab is administered weekly or biweekly, while cobimetinib, or a salt thereof, and palbociclib, or a salt thereof, is administered daily.

[0094] On the other hand, the method comprises administering an effective amount of cetuximab, TAK-733 or a salt thereof, and palbociclib or a salt thereof. In some embodiments, cetuximab is administered before TAK-733 or a salt thereof and palbociclib or a salt thereof. In some embodiments, TAK-733 or a salt thereof is administered before cetuximab and palbociclib or a salt thereof. In some embodiments, palbociclib or a salt thereof is administered before cetuximab and TAK-733 or a salt thereof. In some embodiments, cetuximab and TAK-733 or a salt thereof are administered together in the same or separate compositions. In some embodiments, cetuximab and palbociclib or a salt thereof are administered together in the same or separate compositions. In some embodiments, TAK-733 or a salt thereof and palbociclib or a salt thereof are administered together in the same or separate compositions. In some embodiments, cetuximab is administered as a composition separate from TAK-733 or a salt thereof and palbociclib or a salt thereof. In some embodiments, cetuximab is administered weekly or biweekly, while TAK-733, or a salt thereof, and palbociclib, or a salt thereof, are administered daily.

[0095] On the other hand, the method comprises administering an effective amount of osimertinib or a salt thereof, TAK-733 or a salt thereof, and palbociclib or a salt thereof. In some embodiments, osimertinib or a salt thereof is administered before TAK-733 or a salt thereof and palbociclib or a salt thereof. In some embodiments, TAK-733 or a salt thereof is administered before osimertinib or a salt thereof and palbociclib or a salt thereof. In some embodiments, palbociclib or a salt thereof is administered before osimertinib or a salt thereof and TAK-733 or a salt thereof. In some embodiments, osimertinib or a salt thereof and TAK-733 or a salt thereof are administered together in the same or separate compositions. In some embodiments, osimertinib or a salt thereof and palbociclib or a salt thereof are administered together in the same or separate compositions. In some embodiments, TAK-733 or a salt thereof and palbociclib or a salt thereof are administered together in the same or separate compositions.

[0096] In another aspect, the method comprises administering an effective amount of cetuximab, trametinib, or a salt thereof, and palbociclib, or a salt thereof. In some embodiments, cetuximab is administered before trametinib, or a salt thereof, and palbociclib, or a salt thereof. In some embodiments, trametinib, or a salt thereof, is administered before cetuximab and palbociclib, or a salt thereof. In some embodiments, palbociclib, or a salt thereof, is administered before cetuximab and trametinib, or a salt thereof. In some embodiments, cetuximab and trametinib, or a salt thereof, are administered together in the same or separate compositions. In some embodiments, cetuximab and palbociclib, or a salt thereof, are administered together in the same or separate compositions. In some embodiments, trametinib, or a salt thereof, and palbociclib, or a salt thereof, are administered together in the same or separate compositions. In some embodiments, cetuximab is administered in a composition separate from trametinib, or a salt thereof, and palbociclib, or a salt thereof. In some embodiments, cetuximab is administered weekly or biweekly, while trametinib, or a salt thereof, and palbociclib, or a salt thereof, is administered daily.

[0097] In another aspect, the method comprises administering an effective amount of osimertinib or a salt thereof, trametinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, osimertinib or a salt thereof is administered before trametinib or a salt thereof and palbociclib or a salt thereof. In some embodiments, trametinib or a salt thereof is administered before osimertinib or a salt thereof and palbociclib or a salt thereof. In some embodiments, palbociclib or a salt thereof is administered before osimertinib or a salt thereof and trametinib or a salt thereof. In some embodiments, osimertinib or a salt thereof and trametinib or a salt thereof are administered together in the same or separate compositions. In some embodiments, osimertinib or a salt thereof and palbociclib or a salt thereof are administered together in the same or separate compositions. In some embodiments, trametinib or a salt thereof and palbociclib or a salt thereof are administered together in the same or separate compositions.

[0098] In some embodiments, intermittent administration is at intervals of about 1-30 minutes, about 30-60 minutes, about 60-120 minutes, about 120-240 minutes, about 240-480 minutes, about 480-720 minutes, about 720-960 minutes, or about 960-1440 minutes. In some embodiments, intermittent administration is at intervals of about 1-2 days, 2-3 days, 3-4 days, 4-5 days, 5-6 days, or 6-7 days.

[0099] On the other hand, the subject has previously been treated with a BRAF inhibitor. On the other hand, the subject has not previously been treated with a BRAF inhibitor. On the other hand, the subject has previously been treated with a combination of a BRAF inhibitor and a MEK inhibitor. On the other hand, the subject has not previously been treated with a combination of a BRAF inhibitor and a MEK inhibitor. In some embodiments, the subject has developed acquired or adaptive resistance to a BRAF inhibitor or a MEK inhibitor. In some embodiments, the subject has not yet developed acquired or adaptive resistance to a BRAF inhibitor or a MEK inhibitor. MEK inhibitors include, but are not limited to, small molecules or antibodies that specifically bind to MEK 1 / 2 or its ligands, such as trametinib, selumetinib, TAK-733, CI-1040, PD0325901, MEK162, AZD8330, GDC-0623, remetinib, pimasetinib, RO4987655, RO5126766, WX-554, HL-085, and cobimetinib. BRAF inhibitors include, but are not limited to, small molecules or antibodies that specifically bind to wild-type or mutant BRAF or its ligands, such as vemurafenib and dabrafenib.

[0100] In another aspect, the methods described herein reduce cancer cell growth and / or increase cancer cell killing by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more compared to administration of only one or two of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In another aspect, the methods described herein reduce cancer cell growth and / or increase cancer cell killing by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more compared to administration of only one or both of cetuximab, cobimetinib, or a salt thereof, and palbociclib, or a salt thereof. In another aspect, the methods described herein reduce cancer cell growth and / or increase cancer cell killing by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more compared to administration of only one or two of cetuximab, TAK-733, and palbociclib, or a salt thereof. In another aspect, the methods described herein reduce cancer cell growth and / or increase cancer cell killing by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more compared to administration of only one or two of osimertinib, or a salt thereof, TAK-733, and palbociclib, or a salt thereof. In another aspect, the methods described herein reduce cancer cell growth and / or increase cancer cell killing by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more compared to administration of only one or two of cetuximab, trametinib, and palbociclib, or salts thereof.In another aspect, the methods described herein reduce cancer cell growth and / or increase cancer cell killing by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more compared to administration of only one or two of osimertinib, or a salt thereof, trametinib, and palbociclib, or a salt thereof.

[0101] And the methods described herein have shown synergistic effects in the treatment of cancers with BRAF mutations. In some embodiments, the efficacy of the methods described herein is at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more compared to the additive efficacy of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof administered alone. In some embodiments, the efficacy of the methods described herein is at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more compared to the additive efficacy of cetuximab, cobimetinib, or a salt thereof, and palbociclib, or a salt thereof, administered alone. In some embodiments, the efficacy of the methods described herein is at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more compared to the additive efficacy of cetuximab, TAK-733, or a salt thereof, and palbociclib, or a salt thereof, administered alone. In some embodiments, the efficacy of the methods described herein is at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more compared to the additive efficacy of osimertinib or a salt thereof, TAK-733 or a salt thereof, and palbociclib or a salt thereof administered alone. In some embodiments, the efficacy of the methods described herein is at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more compared to the additive efficacy of cetuximab, trametinib, or a salt thereof, and palbociclib, or a salt thereof, administered alone.In some embodiments, the efficacy of the methods described herein is at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more compared to the additive efficacy of osimertinib, or a salt thereof, trametinib, or a salt thereof, and palbociclib, or a salt thereof, administered alone.

[0102] In another aspect, the methods described herein reduce mean tumor volume. In some embodiments, the methods described herein reduce mean tumor volume by about 20-95%. In some embodiments, mean tumor volume is reduced by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%.

[0103] In another aspect, the methods described herein cause a change in body weight of the subject of less than about 25%, less than about 20%, less than about 15%, or less than about 5%.In some embodiments, the methods do not cause a change in body weight.

[0104] On the other hand, the methods described herein include administering a loading dose of a combination of: (a) an epidermal growth factor receptor (EGFR) inhibitor (e.g., osimertinib or cetuximab); (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor (e.g., cobimetinib, trametinib, or TAK-733); and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor (e.g., palbociclib), followed by multiple separate administrations of the combination at a maintenance dose. In some embodiments, each loading dose of the three inhibitors is higher than each maintenance dose. In some embodiments, the methods described herein provide for long-term administration and / or treatment with lower doses, safety, and / or tolerability.

[0105] III. Composition

[0106] Also provided herein are compositions comprising: (a) an epidermal growth factor receptor (EGFR) inhibitor (e.g., osimertinib or cetuximab); (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor (e.g., cobimetinib, trametinib, or TAK-733); and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor (e.g., palbociclib); wherein the composition does not comprise a BRAF inhibitor. In some embodiments, the composition is composed of: (a) an epidermal growth factor receptor (EGFR) inhibitor (e.g., osimertinib or cetuximab); (b) a mitogen-activated protein kinase (MEK) 1 / 2 inhibitor (e.g., cobimetinib, trametinib, or TAK-733); and (c) a cyclin-dependent kinase (CDK) 4 / 6 inhibitor (e.g., palbociclib).

[0107] Also provided herein are compositions comprising osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. Also provided herein are compositions comprising osimertinib or a salt thereof, TAK-733 or a salt thereof, and palbociclib or a salt thereof. Also provided herein are compositions comprising osimertinib or a salt thereof, trametinib or a salt thereof, and palbociclib or a salt thereof. These compositions can be used in the methods described herein to treat cancers with BRAF mutations and to delay their progression. In some embodiments, the composition does not comprise a BRAF inhibitor.

[0108] In one aspect, the composition may also include a pharmaceutically acceptable carrier, excipient, adhesive or diluent. A pharmaceutically acceptable excipient is a non-toxic and otherwise biologically suitable substance for administering to a subject. Such excipients promote administration of the compounds described herein and are compatible with the active ingredient. Examples of pharmaceutically acceptable excipients include stabilizers, lubricants, surfactants, diluents, antioxidants, adhesives, colorants, fillers, emulsifiers or taste improvers. In some embodiments, the pharmaceutical composition according to the embodiment is a sterile composition. Pharmaceutical compositions can be prepared using known or available dispensing techniques for those skilled in the art. Embodiments also contemplate sterile compositions, including compositions that comply with national and local regulations governing such compositions.

[0109] According to conventional methods for preparing various dosage forms known in the art, the pharmaceutical compositions and compounds described herein can be formulated into solutions, emulsions, suspensions, dispersants or inclusion compounds (such as cyclodextrins) in suitable pharmaceutical solvents or carriers, or formulated into pills, tablets, lozenges, suppositories, sachets, dragees, granules, powders, powders for reconstitution or capsules together with solid carriers. The pharmaceutical compositions provided herein can be administered by suitable delivery routes, such as oral, parenteral, rectal, nasal or topical routes, or by inhalation. In some embodiments, the compositions are formulated for intravenous or oral administration.

[0110] On the other hand, the composition is formulated for oral administration. For oral administration, the composition can be formulated into solid forms, such as tablets or capsules, or formulated into solutions, emulsions or suspensions. Oral tablets can comprise one or more active ingredients mixed with compatible pharmaceutically acceptable excipients (such as diluents, disintegrants, adhesives, lubricants, sweeteners, flavorings, coloring agents and preservatives). Suitable inert fillers include sodium carbonate and calcium carbonate, sodium phosphate and calcium phosphate, lactose, starch, sugar, glucose, methylcellulose, magnesium stearate, mannitol, sorbitol etc. Exemplary liquid oral excipients include ethanol, glycerol, water etc. Starch, polyvinyl pyrrolidone (PVP), sodium starch glycolate, microcrystalline cellulose and alginic acid are exemplary disintegrants. Adhesives can include starch and gelatin. If present, lubricants can be magnesium stearate, stearic acid or talc. If desired, tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate to delay absorption in the gastrointestinal tract, or may be provided with an enteric coating.

[0111] Capsules for oral administration include hard and soft gelatin capsules. In order to prepare hard gelatin capsules, one or more active ingredients can be mixed with a solid, semisolid or liquid diluent. Soft gelatin capsules can be prepared by mixing the active ingredient with water, oil (such as peanut oil or olive oil), liquid paraffin, a mixture of monoglycerides and diglycerides of short-chain fatty acids, polyethylene glycol 400 or propylene glycol.

[0112] Liquids for oral administration may be in the form of suspensions, solutions, emulsions, or syrups, or may be lyophilized or presented as dry products for reconstitution with water or other suitable vehicles before use. Such liquid compositions may optionally contain: pharmaceutically acceptable excipients, such as suspending agents (e.g., sorbitol, methylcellulose, sodium alginate, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel, etc.); non-aqueous vehicles, such as oils (e.g., almond oil or fractionated coconut oil), propylene glycol, ethanol, or water; preservatives (e.g., methylparaben or propylparaben or sorbic acid); wetting agents, such as lecithin; and, if desired, flavorings or coloring agents.

[0113] In another aspect, the composition comprises osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the composition comprises greater than about 20%, or greater than about 25%, or greater than about 30%, or greater than about 35%, or greater than about 40%, or greater than about 45%, or greater than about 50%, or greater than about 55%, or greater than about 60%, or greater than about 65%, or greater than about 70% by weight of osimertinib or a salt thereof.

[0114] In some embodiments, the composition comprises greater than about 20%, or greater than about 25%, or greater than about 30%, or greater than about 35%, or greater than about 40%, or greater than about 45%, or greater than about 50% by weight of cobicistinib or a salt thereof.

[0115] In some embodiments, the composition comprises greater than about 30%, or greater than about 35%, or greater than about 40%, or greater than about 45%, or greater than about 50%, or greater than about 55%, or greater than about 60%, or greater than about 65%, or greater than about 70%, or greater than about 75%, or greater than about 80%, or greater than about 85%, or greater than about 90% palbociclib or a salt thereof by weight.

[0116] In some embodiments, the ratio of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof in the composition by weight is about 1:1:1, 2:1:1, 3:1:1, 4:1:1, 1:2:1, 1:1:2, 1:1:3, 1:1:4, 1:1:5, 1:1:6, 1:1:7, 1:1:8, 1:1:9, 1:1:10, 2:1:2, 2:1:3, 2:1:4, 2:1:5, 2:1:6, 2:1:7, 2:1:8, 2:1:9, 2:1:10, 3:1:1, 3:1:2, 3:1:3, 3:1:4, 3:1:5, 3:1:6, 3:1:7, 3:1:8, 3:1:9, or 3:1:10. In some embodiments, the ratio of osimertinib or its salt, cobimetinib or its salt, and palbociclib or its salt in the composition is about 2:1:4 by weight. In some embodiments, the ratio of osimertinib or its salt and cobimetinib or its salt is in the range of about 2:3 to 4:1 by weight. In some embodiments, the ratio of osimertinib or its salt and palbociclib or its salt is in the range of about 40:125 to 80:75 by weight. In some embodiments, the ratio of cobimetinib or its salt and palbociclib or its salt is in the range of about 20:125 to 60:75 by weight.

[0117] In another aspect, the composition comprises a salt of osimertinib, cobimetinib, TAK-733, trametinib, or palbociclib. In some embodiments, the salt is a pharmaceutically acceptable salt. Non-limiting examples of pharmaceutically acceptable salts include, but are not limited to, sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, octanoates, acrylates, formates, isobutyrates, hexanoates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyrates, succinate ... Alkyne-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, methylsulfonate, propylsulfonate, benzenesulfonate, xylenesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, phenylacetate, methanesulfonate phenylpropionate, phenylbutyrate, citrate, lactate, γ-hydroxybutyrate, glycolate, tartrate and mandelate. In some embodiments, the composition comprises the mesylate of osimertinib, cobimetinib and palbociclib. In some embodiments, the composition comprises the fumarate of osimertinib, cobimetinib and palbociclib. On the other hand, the composition comprises a solvate of osimertinib, cobimetinib, TAK-722 or palbociclib.

[0118] In another aspect, the composition comprises osimertinib or a salt thereof, TAK-733 or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the composition comprises greater than about 20%, or greater than about 25%, or greater than about 30%, or greater than about 35%, or greater than about 40%, or greater than about 45%, or greater than about 50%, or greater than about 55%, or greater than about 60%, or greater than about 65%, or greater than about 70% by weight of osimertinib or a salt thereof.

[0119] In some embodiments, the composition comprises greater than about 20%, or greater than about 25%, or greater than about 30%, or greater than about 35%, or greater than about 40%, or greater than about 45%, or greater than about 50% by weight of TAK-733 or a salt thereof.

[0120] In some embodiments, the composition comprises greater than about 30%, or greater than about 35%, or greater than about 40%, or greater than about 45%, or greater than about 50%, or greater than about 55%, or greater than about 60%, or greater than about 65%, or greater than about 70%, or greater than about 75%, or greater than about 80%, or greater than about 85%, or greater than about 90% palbociclib or a salt thereof by weight.

[0121] In some embodiments, the ratio of osimertinib or a salt thereof, TAK-733 or a salt thereof, and palbociclib or a salt thereof by weight in the composition is about 1:1:1, 2:1:1, 3:1:1, 4:1:1, 1:2:1, 1:1:2, 1:1:3, 1:1:4, 1:1:5, 1:1:6, 1:1:7, 1:1:8, 1:1:9, 1:1:10, 2:1:2, 2:1:3, 2:1:4, 2:1:5, 2:1:6, 2:1:7, 2:1:8, 2:1:9, 2:1:10, 3:1:1, 3:1:2, 3:1:3, 3:1:4, 3:1:5, 3:1:6, 3:1:7, 3:1:8, 3:1:9, or 3:1:10. In some embodiments, the ratio of osimertinib or its salt to TAK-733 or its salt in the composition is in the range of about 5:2 to 10:1 by weight. In some embodiments, the ratio of osimertinib or its salt to palbociclib or its salt in the composition is in the range of about 40:125 to 80:75 by weight. In some embodiments, the ratio of TAK-733 or its salt to palbociclib or its salt in the composition is in the range of about 8:125 to 16:75 by weight.

[0122] In another aspect, the composition comprises osimertinib or a salt thereof, trametinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the composition comprises greater than about 20%, or greater than about 25%, or greater than about 30%, or greater than about 35%, or greater than about 40%, or greater than about 45%, or greater than about 50%, or greater than about 55%, or greater than about 60%, or greater than about 65%, or greater than about 70% by weight of osimertinib or a salt thereof.

[0123] In some embodiments, the composition comprises greater than about 20%, or greater than about 25%, or greater than about 30%, or greater than about 35%, or greater than about 40%, or greater than about 45%, or greater than about 50% by weight of trametinib or a salt thereof.

[0124] In some embodiments, the composition comprises greater than about 30%, or greater than about 35%, or greater than about 40%, or greater than about 45%, or greater than about 50%, or greater than about 55%, or greater than about 60%, or greater than about 65%, or greater than about 70%, or greater than about 75%, or greater than about 80%, or greater than about 85%, or greater than about 90% palbociclib or a salt thereof by weight.

[0125] In some embodiments, the ratio of osimertinib or a salt thereof, trametinib or a salt thereof, and palbociclib or a salt thereof in the composition by weight is about 15:1:10, 20:1:10, 25:1:10, 30:1:10, 35:1:10, 40:1:10, 50:1:10, 15:1:20, 20:1:20, 25:1:20, 30:1:20, 35:1:20, 40:1:20, 50:1:20, 15:1:30, 20:1:30, 25:1:30, 30:1:30, 35:1:30, 40:1:30, 50:1:30, or 40:1:50. In some embodiments, the ratio of osimertinib or its salt, trametinib or its salt, and palbociclib or its salt in the composition is about 15:1:12 by weight. In some embodiments, the ratio of osimertinib or its salt and trametinib or its salt in the composition is in the range of about 20:1 to 160:1 by weight. In some embodiments, the ratio of osimertinib or its salt and palbociclib or its salt in the composition is in the range of about 40:125 to 80:75 by weight. In some embodiments, the ratio of trametinib or its salt and palbociclib or its salt in the composition is in the range of about 1:250 to 2:75 by weight.

[0126] IV. Kit

[0127] Also provided herein is a kit comprising the following: (a) epidermal growth factor receptor (EGFR) inhibitor (for example, osimertinib or cetuximab); (b) mitogen-activated protein kinase (MEK) 1 / 2 inhibitor (for example, TAK-733, trametinib or cobimetinib); and (c) cyclin-dependent kinase (CDK) 4 / 6 inhibitor (for example, palbociclib); wherein the kit does not include BRAF inhibitors. In some embodiments, the kit is composed of the following: (a) epidermal growth factor receptor (EGFR) inhibitor (for example, osimertinib or cetuximab); (b) mitogen-activated protein kinase (MEK) 1 / 2 inhibitor (for example, TAK-733, trametinib or cobimetinib); and (c) cyclin-dependent kinase (CDK) 4 / 6 inhibitor (for example, palbociclib). The kit can be used in the methods described herein for treating cancer with BRAF mutations and delaying their progression. In some embodiments, the kit does not include BRAF inhibitors. The kit may comprise any of the compositions described herein.

[0128] Also provided herein are kits comprising osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the kit comprises a pharmaceutical composition comprising osimertinib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising cobimetinib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising palbociclib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent.

[0129] In some embodiments, osimertinib or its salt, cobimetinib or its salt and palbociclib or its salt are formulated into a composition in a kit. In some embodiments, osimertinib or its salt, cobimetinib or its salt and palbociclib or its salt are formulated separately. In some embodiments, osimertinib or its salt, cobimetinib or its salt and / or palbociclib or its salt are formulated for oral administration. In some embodiments, osimertinib or its salt, cobimetinib or its salt and palbociclib or its salt are formulated into the same form, such as solid or liquid form. In some embodiments, osimertinib or its salt, cobimetinib or its salt and palbociclib or its salt are formulated into a solution, emulsion, suspension, dispersant or inclusion compound (such as cyclodextrin) in a suitable pharmaceutical solvent or carrier according to conventional methods for preparing various dosage forms known in the art, or formulated into pills, tablets, lozenges, suppositories, sachets, dragees, granules, powders, powders for reconstitution or capsules together with a solid carrier.

[0130] In some embodiments, the kit provides osimertinib, or a salt thereof, cobimetinib, or a salt thereof, and palbociclib, or a salt thereof, in a ratio by weight of about 1:1:1, 2:1:1, 3:1:1, 4:1:1, 1:2:1, 1:1:2, 1:1:3, 1:1:4, 1:1:5, 1:1:6, 1:1:7, 1:1:8, 1:1:9, 1:1:10, 2:1:2, 2:1:3, 2:1:4, 2:1:5, 2:1:6, 2:1:7, 2:1:8, 2:1:9, 2:1:10, 3:1:1, 3:1:2, 3:1:3, 3:1:4, 3:1:5, 3:1:6, 3:1:7, 3:1:8, 3:1:9, or 3:1:10. In some embodiments, the ratio of osimertinib or a salt thereof, cobimetinib or a salt thereof, and palbociclib or a salt thereof in the kit by weight is about 2:1:4. In some embodiments, the ratio of osimertinib or a salt thereof and cobimetinib or a salt thereof in the kit by weight is in the range of about 2:3 to 4:1. In some embodiments, the ratio of osimertinib or a salt thereof and palbociclib or a salt thereof in the kit by weight is in the range of about 40:125 to 80:75. In some embodiments, the ratio of cobimetinib or a salt thereof and palbociclib or a salt thereof in the kit by weight is in the range of about 20:125 to 60:75.

[0131] Also provided herein are kits comprising cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof. In some embodiments, the kit comprises a pharmaceutical composition comprising cetuximab and a pharmaceutically acceptable carrier, excipient, binder or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising cobimetinib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising palbociclib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder or diluent.

[0132] In some embodiments, cetuximab, cobimetinib or its salt and palbociclib or its salt are formulated into two or more compositions in a kit. In some embodiments, cetuximab, cobimetinib or its salt and palbociclib or its salt are formulated separately. In some embodiments, cobimetinib or its salt and / or palbociclib or its salt are formulated for oral administration. In some embodiments, cobimetinib or its salt and palbociclib or its salt are formulated into the same form, such as solid or liquid form. In some embodiments, according to conventional methods for preparing various dosage forms known in the art, cetuximab, cobimetinib or its salt and / or palbociclib or its salt are formulated into solutions, emulsions, suspensions, dispersants or inclusion compounds (such as cyclodextrins) in suitable pharmaceutical solvents or carriers, or are formulated into pills, tablets, lozenges, suppositories, sachets, dragees, granules, powders, powders for reconstruction or capsules together with a solid carrier. In some embodiments, cetuximab is formulated for administration via intravenous infusion. In some embodiments, cobimetinib or a salt thereof and palbociclib or a salt thereof are formulated as a composition for oral administration.

[0133] In some embodiments, the kit provides a ratio of cetuximab, cobimetinib, or a salt thereof, and palbociclib, or a salt thereof, by weight of about 1:1:1:, 2:1:1, 3:1:1, 4:1:1:, 1:2:1, 3:1:2, 3:1:3, 3:1:4, 3:1:5, 3:1:6, 3:1:7, 3:1:8, 3:1:9, 3:1:10, 4:1:2, 4:1:3, 4:1:4, 4:1:5. In some embodiments, the ratio of cetuximab, cobimetinib or a salt thereof and palbociclib or a salt thereof in the kit is about 20:3:6 by weight. In some embodiments, the ratio of cetuximab and cobimetinib or a salt thereof in the kit is about 85:60 to 85:2 by weight. In some embodiments, the ratio of cetuximab to palbociclib or its salt in the kit by weight is in the range of about 17:25 to 34:3. In some embodiments, the ratio of cobimetinib or its salt to palbociclib or its salt in the kit by weight is in the range of about 20:125 to 60:75.

[0134] Also provided herein are kits comprising cetuximab, TAK-733 or a salt thereof and palbociclib or a salt thereof. In some embodiments, the kit comprises a pharmaceutical composition comprising cetuximab and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising TAK-733 or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising palbociclib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent.

[0135] In some embodiments, cetuximab, TAK-733 or its salt and palbociclib or its salt are formulated into a composition in a kit. In some embodiments, cetuximab, TAK-733 or its salt and palbociclib or its salt are formulated into two or more compositions in a kit. In some embodiments, cetuximab, TAK-733 or its salt and palbociclib or its salt are formulated separately. In some embodiments, TAK-733 or its salt and / or palbociclib or its salt are formulated for oral administration. In some embodiments, TAK-733 or its salt and palbociclib or its salt are formulated into the same form, such as solid or liquid form. In some embodiments, cetuximab, TAK-733 or its salt and / or palbociclib or its salt are formulated into solutions, emulsions, suspensions, dispersions or inclusion compounds (such as cyclodextrins) in suitable pharmaceutical solvents or carriers according to conventional methods for preparing various dosage forms known in the art, or are formulated into pills, tablets, lozenges, suppositories, sachets, dragees, granules, powders, powders for reconstitution or capsules together with solid carriers. In some embodiments, cetuximab is formulated for administration via intravenous infusion. In some embodiments, TAK-733 or its salt and palbociclib or its salt are formulated into a composition for oral administration.

[0136] In some embodiments, the kit provides a ratio of cetuximab, TAK-733 or a salt thereof, and palbociclib or a salt thereof by weight of about 5:1:5:, 10:1:5, 15:1:5, 20:1:5:, 10:2:1, 10:1:2, 10:1:3, 10:1:4, 10:1:5, 10:1:6, 10:1:7, 10:1:8, 5:1:9, 10:1:10, 10:1:11, 10:1:12, 10:1:13, 10:1:14, 10:1:15, 10:1:16, 10:1:17, 10:1:18, 10:1:19, 10:1:11, 10:1:12, 1 In some embodiments, the ratio of cetuximab to TAK-733 or a salt thereof and palbociclib or a salt thereof by weight in the kit is about 20:1:6. In some embodiments, the ratio of cetuximab to TAK-733 or a salt thereof by weight in the kit is in the range of about 85:16 to 850:8. In some embodiments, the ratio of cetuximab to palbociclib or its salt in the kit by weight is in the range of about 17:25 to 34:3. In some embodiments, the ratio of TAK-733 or its salt to palbociclib or its salt in the kit by weight is in the range of about 8:125 to 16:75.

[0137] Also provided herein are kits comprising osimertinib or a salt thereof, TAK-733 or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the kit comprises a pharmaceutical composition comprising osimertinib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising TAK-733 or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising palbociclib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent.

[0138] In some embodiments, osimertinib or a salt thereof, TAK-733 or a salt thereof, and palbociclib or a salt thereof are formulated into a composition in a kit. In some embodiments, osimertinib or a salt thereof, TAK-733 or a salt thereof, and palbociclib or a salt thereof are formulated separately. In some embodiments, osimertinib or a salt thereof, TAK-733 or a salt thereof, and / or palbociclib or a salt thereof are formulated for oral administration. In some embodiments, osimertinib or a salt thereof, TAK-733 or a salt thereof, and palbociclib or a salt thereof are formulated into the same form, such as a solid or liquid form. In some embodiments, osimertinib or its salt, TAK-733 or its salt and / or palbociclib or its salt are formulated into solutions, emulsions, suspensions, dispersions or inclusion compounds (such as cyclodextrin) in suitable pharmaceutical solvents or carriers according to conventional methods for preparing various dosage forms known in the art, or formulated into pills, tablets, lozenges, suppositories, sachets, dragees, granules, powders, powders for reconstitution or capsules together with solid carriers.

[0139] In some embodiments, the kit provides osimertinib, or a salt thereof, TAK-733, or a salt thereof, and palbociclib, or a salt thereof, in a ratio by weight of about 1:1:1, 2:1:1, 3:1:1, 4:1:1, 1:2:1, 1:1:2, 1:1:3, 1:1:4, 1:1:5, 1:1:6, 1:1:7, 1:1:8, 1:1:9, 1:1:10, 2:1:2, 2:1:3, 2:1:4, 2:1:5, 2:1:6, 2:1:7, 2:1:8, 2:1:9, 2:1:10, 3:1:1, 3:1:2, 3:1:3, 3:1:4, 3:1:5, 3:1:6, 3:1:7, 3:1:8, 3:1:9, or 3:1:10. In some embodiments, the ratio of osimertinib or its salt, TAK-733 or its salt, and palbociclib or its salt in the composition is about 3:1:6 by weight. In some embodiments, the ratio of osimertinib or its salt and TAK-733 or its salt in the kit is in the range of about 5:2 to 10:1 by weight. In some embodiments, the ratio of osimertinib or its salt and palbociclib or its salt in the kit is in the range of about 40:125 to 80:75 by weight. In some embodiments, the ratio of TAK-733 or its salt and palbociclib or its salt in the kit is in the range of about 8:125 to 16:75 by weight.

[0140] Also provided herein are kits comprising cetuximab, trametinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the kit comprises a pharmaceutical composition comprising cetuximab and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising trametinib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising palbociclib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent.

[0141] In some embodiments, cetuximab, trametinib or a salt thereof, and palbociclib or a salt thereof are formulated into one composition in a kit. In some embodiments, cetuximab, trametinib or a salt thereof, and palbociclib or a salt thereof are formulated into two or more compositions in a kit. In some embodiments, cetuximab, trametinib or a salt thereof, and palbociclib or a salt thereof are formulated separately. In some embodiments, trametinib or a salt thereof and / or palbociclib or a salt thereof are formulated for oral administration. In some embodiments, trametinib or a salt thereof and palbociclib or a salt thereof are formulated into the same form, such as a solid or liquid form. In some embodiments, cetuximab, trametinib or its salt and / or palbociclib or its salt are formulated into solutions, emulsions, suspensions, dispersions or inclusion compounds (such as cyclodextrins) in suitable pharmaceutical solvents or carriers according to conventional methods for preparing various dosage forms known in the art, or formulated into pills, tablets, lozenges, suppositories, sachets, dragees, granules, powders, powders for reconstitution or capsules together with solid carriers. In some embodiments, cetuximab is formulated for administration via intravenous infusion. In some embodiments, trametinib or its salt and palbociclib or its salt are formulated into a composition for oral administration.

[0142] Also provided herein are kits comprising osimertinib or a salt thereof, trametinib or a salt thereof, and palbociclib or a salt thereof. In some embodiments, the kit comprises a pharmaceutical composition comprising osimertinib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising trametinib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent. In some embodiments, the kit comprises a pharmaceutical composition comprising palbociclib or a salt thereof and a pharmaceutically acceptable carrier, excipient, binder, or diluent.

[0143] In some embodiments, osimertinib or its salt, trametinib or its salt and palbociclib or its salt are formulated into a composition in a kit. In some embodiments, osimertinib or its salt, trametinib or its salt and palbociclib or its salt are formulated separately. In some embodiments, osimertinib or its salt, trametinib or its salt and / or palbociclib or its salt are formulated for oral administration. In some embodiments, osimertinib or its salt, trametinib or its salt and palbociclib or its salt are formulated into the same form, such as solid or liquid form. In some embodiments, osimertinib or its salt, trametinib or its salt and palbociclib or its salt are formulated into a solution, emulsion, suspension, dispersant or inclusion compound (such as cyclodextrin) in a suitable pharmaceutical solvent or carrier according to conventional methods for preparing various dosage forms known in the art, or formulated into pills, tablets, lozenges, suppositories, sachets, dragees, granules, powders, powders for reconstitution or capsules together with a solid carrier.

[0144] In some embodiments, the kit provides osimertinib, or a salt thereof, trametinib, or a salt thereof, and palbociclib, or a salt thereof, in a ratio by weight of about 15:1:10, 20:1:10, 25:1:10, 30:1:10, 35:1:10, 40:1:10, 50:1:10, 15:1:20, 20:1:20, 25:1:20, 30:1:20, 35:1:20, 40:1:20, 50:1:20, 15:1:30, 20:1:30, 25:1:30, 30:1:30, 35:1:30, 40:1:30, 50:1:30, or 40:1:50. In some embodiments, the ratio of osimertinib or its salt, trametinib or its salt, and palbociclib or its salt in the composition is about 15:1:12 by weight. In some embodiments, the ratio of osimertinib or its salt and trametinib or its salt in the kit is in the range of about 20:1 to 160:1 by weight. In some embodiments, the ratio of osimertinib or its salt and palbociclib or its salt in the kit is in the range of about 40:125 to 80:75 by weight. In some embodiments, the ratio of trametinib or its salt and palbociclib or its salt in the kit is in the range of about 1:250 to 2:75 by weight.

[0145] In another aspect, the kit further comprises a package insert including, but not limited to, appropriate instructions for preparing and administering the formulation, side effects of the formulation, and any other relevant information. The instructions may be in any suitable format, including but not limited to printed matter, videotape, computer readable disk, CD-ROM, or instructions based on the internet.

[0146] In another aspect, a kit for treating an individual suffering from or susceptible to a condition described herein is provided, comprising a first container containing a dose of a composition or formulation as disclosed herein and a package insert for use. The container can be any container known in the art suitable for storing and delivering intravenous formulations. In certain embodiments, the kit further comprises a second container containing a pharmaceutically acceptable carrier, diluent, adjuvant, etc. for preparing the formulation to be administered to the individual.

[0147] In another aspect, kits can also be provided that contain sufficient doses of the compositions described herein (including pharmaceutical compositions thereof) to provide effective treatment to an individual for an extended period of time (such as 1-3 days, 1-5 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 1 cycle, 2 cycles, 3 cycles, 4 cycles, 5 cycles, 6 cycles, 7 cycles, 8 cycles or longer).

[0148] In some embodiments, the kit may also include multiple doses and may be packaged in amounts sufficient for storage and use in pharmacies (e.g., hospital pharmacies and compounding pharmacies). In certain embodiments, the kit may include a dose of at least one composition as disclosed herein.

[0149] Example

[0150] The following examples are provided to illustrate, but not to limit, the present invention. Those skilled in the art will recognize that the following procedures can be modified using methods known to those of ordinary skill in the art.

[0151] Example 1 In vitro drug testing using conditionally reprogrammed cell pools

[0152] Surgical specimens from patients with colorectal cancer were obtained from hospitals in Beijing after obtaining informed consent from the patients. Patient-derived xenograft tumor samples were obtained from Nod / SCID mice inoculated with the patients' surgical tumor specimens.

[0153] For ex vivo drug sensitivity assays, colorectal tumor cells (CRC) were isolated from patient tissue samples or PDX xenograft tumor tissue. Briefly, tumor tissue was minced into small pieces less than 1 mm in diameter using scissors. The tumor fragments were transferred to a sterile 100 ml triangular glass flask equipped with a magnetic stir bar. 10-15 ml of digestion medium containing 0.25 U / ml Liberase DH was added to the minced tumor tissue to initiate enzymatic digestion. The enzyme mixture was incubated at 37°C for 1-2 hours with moderate stirring. The digested tumor tissue was filtered through a 100 μm cell holder. The filtrate was re-filtered through a 40 μm cell holder. The CRC mass retained on the 40 μm cell holder was collected, washed twice with HBSS, and then resuspended in a defined growth medium supplemented with several stem cell growth factors.

[0154] CRC pellets retained on 40-μm cell holders were collected, washed twice with HBSS, and then resuspended in defined growth medium supplemented with cell growth factors and small molecule inhibitors. CRC pellets were recovered overnight in defined growth medium. Defined growth medium is hESC SFM (defined serum- and feeder-free medium (SFM)) supplemented with: Nicotinamide, Wnt3A, Noggin (bone morphogenetic protein (BMP) inhibitor), Rspondin-1 (Wnt / β-catenin signaling agonist), and Y27632 (Rho-associated coiled-coil-containing protein kinase (ROCK-1) inhibitor).

[0155] For conditional reprogramming, recovered cancer tissue-derived spheroids (CTOS) were dissociated into single cells and seeded at approximately 3,000 cells per well in a feeder cell / Rock inhibitor (Y27632) co-culture system in cell culture plates and grown at 37°C for 3 days.

[0156] For in vitro drug testing, seeded CRC cells were exposed to a combination of osimertinib (mesylate), cobimetinib (fumarate), and palbociclib (fumarate) for 72 hours. The combination was prepared in DMSO, and the final DMSO concentration in the culture medium was 0.1%. Typical compound concentrations used for in vitro drug testing were: osimertinib 0.1-0.5 μM, cobimetinib 0.025-0.2 μM, and palbociclib 0.125-1 μM.

[0157] After drug exposure, CRCs were labeled with 5-ethynyl-2'-deoxyuridine (Edu) to assess tumor cell proliferation rate. Labeling continued for 24 hours in the presence of drug exposure. In the control group, epithelial tumor cells were not drug exposed with replacement of the culture medium but were similarly labeled with Edu. The labeled CRCs were fixed in blocking buffer containing 0.5% Triton X-100 and 3% BSA at 4°C and stained with Hoechst 33342 overnight. Tumor cells were incubated with EpCAM antibody (1:4000) for 2 hours at room temperature and rinsed with PBST. Subsequently, CRCs were incubated with Alexa Fluor 500 antibody. The cells were incubated with 647-conjugated goat anti-mouse secondary antibody at room temperature for 30 minutes and then washed with PBST.

[0158] Incorporated Edu was detected by the Click-iT reaction, in which fixed cells were incubated in the dark with a reaction mixture containing 1X Click-iT Edu reaction buffer, CuSO4, and azide-conjugated Alexa Fluor dye. The stained cells were washed twice with PBS before image acquisition and analysis.

[0159] For image acquisition and analysis, the stained tumor cells were imaged using a high content screening (HCS) platform (ThermoScientific CellomicsArrayScanXTi HCS reader). Images were collected using a 10x objective lens. Twenty-five fields of view were imaged for each well in the analysis. From the images, three fluorescent signals were obtained from the HCS reader. The blue fluorescent signal records the signal of the cell nuclei stained with Hoechst 33342, the green fluorescent signal detects Edu incorporated into the newly synthesized DNA, and the red fluorescent signal detects the EpCAM-positive epithelial cell population. Representative images are shown in Figure 1 middle.

[0160] MI (maximum inhibition index) was calculated using EpCAM and Edu positive readings (Table 1). MI = Edu positive cells in control / Edu positive cells in treatment. As shown in Table 1, the combination therapy had the highest specificity for CRC with BRAF V600E or D581V mutations.

[0161] Table 1. Maximum inhibitory index (MI) of CRC treated with combination therapy

[0162] CRC patient ID KRAS mutations BRAF mutation NRAS mutations MI CKY041 G12D 25 NYL170 G12D 39 NYL-JN-025 G12D 106 ZKB171 G12D 908 CKY048 G13D 3898.57 NYL170(NYP023) G12D 39 NYL102 309 NYL109 1085 NYL161 175 NYL-GZ-076 73.67 NYL-HEB-057 G12D 894.06 NYP066 20.87 NYP069 137.86 NYL132 V600E 17882 ZKB197 V600E 10448 ZKB040 D581V 11311 NYL243 V600E 89

[0163] Table 2 shows the selectivity of the combination therapy for CRC with or without BRAF mutation. Selectivity (%) = number of CRCs with MI>=1000 / total number of CRCs*100%

[0164] Table 2 Selectivity of combination therapy for BRAF mutations in CRC

[0165]

[0166] The data presented in Table 2 show that colorectal cancer cells with BRAF mutations were extremely sensitive to the drug combination, with 3 of 4 cells having a greater than 10,000-fold reduction in cell proliferation compared to no-drug controls. In contrast, 2 of 11 colorectal cancer cells without BRAF mutations or KRAS / NRAS mutations had a greater than 1,000-fold reduction in cell proliferation, and none had a greater than 10,000-fold reduction.

[0167] Example 2 Testing of Mouse Tumor Volume Reduction During Combination Therapy in a PDX Model Using Patient NYL132

[0168] The studies were performed in xenograft tumor PDX models using BRAF V600E - Surgical tumor tissue establishment of CRC patient NYL132. The average tumor volume during the combination therapy is shown in Table 3 and Figure 2 Briefly, female NOD / SCID mice aged 6-8 weeks were used for the study. Tumor samples obtained from patients were immediately transferred to tissue preservation fluid and cut into small fragments. The fragments were inoculated subcutaneously on one side of the mouse to produce a xenograft called passage 1 (P1). Serial xenografts of different generations were generated using the same procedure. When the average tumor size of the mice reached approximately 250-300 mm 3 At the same time, animals were randomly assigned to different groups with 5 mice in each group. The date of randomization was defined as study day 0. Tumor volume was expressed in mm using the following formula: 3 V (volume) = (a × b 2 ) / 2, where a and b are the major and minor diameters of the tumor, respectively. Tumor inhibition was expressed as tumor growth inhibition (TGI) calculated according to the following formula: TGI=(1-(Ti-T0) / (Vi-V0))*100%, where Ti is the average tumor volume of the treatment group on the measurement day; T0 is the average tumor volume of the treatment group at D0; Vi is the average tumor volume of the control group on the measurement day; V0 is the tumor volume of the control group at D0. The TGI values ​​of the combination therapy are shown in Tables 4 and Figure 3 middle.

[0169] Table 3 Mean tumor volume during combination therapy

[0170]

[0171] Table 4 Tumor growth inhibition during combination therapy

[0172]

[0173]

[0174] The data presented in Tables 3 and 4 show that the combination therapy reduced tumor volume in a dose-dependent manner. The data demonstrate the critical value of the combination therapy, demonstrating the ability to cause regression of established tumors (not just slowing tumor growth), whereas standard chemotherapy regimens for colorectal cancer are completely ineffective in BRAF mutant colorectal tumor models.

[0175] Example 3 Body weight of mice during combination therapy in a PDX model using patient NYL132

[0176] Use of BRAF V600E -CRC patient NYL132, followed the same procedure as in Example 2. The average body weights of the mice are shown in Tables 5 and Figure 4 middle.

[0177] Table 5 Body weight of mice during combination therapy

[0178]

[0179]

[0180]

[0181] The data presented in Table 4 show that the combination therapy is tolerable in animals, especially at dose levels that show significant in vivo efficacy.

[0182] Example 4. Testing of tumor volume reduction and body weight changes in mice during combination therapy in a PDX model using patient NYL-GZ-082

[0183] Use of BRAF V600E -CRC patient NYL-GZ-082, using various combinations, followed the same procedures as in Examples 2 and 3. Tumor volume values ​​and weight changes are shown in Figure 5 and Figure 6 Both combination therapies: osimertinib + cobimetinib + palbociclib and cetuximab + cobimetinib + palbociclib reduced tumor volume. In addition, in the control group ( Figure 5 , triangles), tumor volume increased to >900 mm after 14 days without treatment 3 Combination therapy (cetuximab + cobimetinib + palbociclib) was administered starting on day 15, and tumor volume decreased rapidly, even after the tumors had grown to a large size. Furthermore, wet weight maintained tumor regression over two treatment cycles.

[0184] Example 5. BRAF treated with monotherapy and combination therapy in a PDX model using patient NYL132 V600E -Comparison of Maximum Inhibitory Index (MI) and Tumor Growth Inhibition (TGI) in CRC

[0185] Use of BRAF V600E -CRC patient NYL132 followed the same procedure as in Example 2. MI (maximum inhibition index) was calculated using EpCAM and Edu-positive readings (Table 6). MI = Edu-positive cells in control / Edu-positive cells in treatment. Tumor growth inhibition (TGI) was calculated according to the following formula: TGI = (1-(Ti-T0) / (Vi-V0))*100%, where Ti is the mean tumor volume of the treatment group on the measurement day; T0 is the mean tumor volume of the treatment group at D0; Vi is the mean tumor volume of the control group on the measurement day; V0 is the tumor volume of the control group at D0.

[0186] Table 6. BRAF treated with monotherapy and combination therapy in PDX models using patient NYL132 V600E - MI and TGI values ​​of CRC

[0187]

[0188]

[0189] Figure 7 Figure 7 shows the changes in tumor volume during treatment with various monotherapy and combination therapy in the PDX model using patient NYL132. All three combinations tested (1) cetuximab + cobimetinib + palbociclib, (2) osimertinib + TAK-733 + palbociclib and (3) cetuximab + TAK-733 + palbociclib reduced tumor volume by >85% in the first treatment cycle (21 days) (see also Table 7). Single cetuximab treatment only slightly inhibited tumor growth (open triangles, 2-cycle treatment from start to day 56), and single cetuximab-treated tumors were sensitive to treatment with a combination of cetuximab 1 mg weekly (ip), cobimetinib 5 mg / kg and palbociclib 20 mg / kg daily (po) for 21 days.

[0190] Table 7. Derived from BRAF V600E -Tumor growth inhibition in the PDX model of CRC patient NYL132

[0191]

[0192]

[0193] Example 6. BRAF treated with monotherapy and combination therapy in a CDX model using the cell line HT-29 V600E -Comparison of Maximum Inhibitory Index (MI) and Tumor Growth Inhibition (TGI) in CRC

[0194] In vivo efficacy studies of xenografts (CDX) derived from the cell line HT-29 were performed. The experimental procedure is briefly described below. Sufficient donor cells (HT-29) were cultured and prepared for seeding. A minimum of 3x10 6 The cells were injected into the dorsal flank of female nude mice. When the tumor volume of the mice grew to 100-300 mm 3 During the study, mice with irregularly shaped tumors and poor health were excluded based on their weight and tumor growth, and the remaining mice were randomly divided into multiple groups, usually 3-5 mice per group. The date of randomization was defined as study day 0. Tumor volume was expressed in mm using the following formula: 3 V (volume) = (a × b 2 ) / 2, where a and b are the major and minor diameters of the tumor, respectively. Tumor inhibition was expressed as tumor growth inhibition (TGI) calculated according to the following formula: TGI = (1-(Ti-T0) / (Vi-V0))*100%, where Ti is the mean tumor volume of the treatment group on the measurement day; T0 is the mean tumor volume of the treatment group at D0; Vi is the mean tumor volume of the control group on the measurement day; V0 is the tumor volume of the control group at D0.

[0195] MI (maximum inhibition index) was calculated using EpCAM and Edu positive readings (Table 8). MI = Edu positive cells in control / Edu positive cells in treatment. Tumor growth inhibition (TGI) was calculated according to the following formula: TGI = (1-(Ti-T0) / (Vi-V0))*100%, where Ti is the mean tumor volume of the treatment group on the measurement day; T0 is the mean tumor volume of the treatment group at D0; Vi is the mean tumor volume of the control group on the measurement day; V0 is the tumor volume of the control group at D0.

[0196] Table 8. BRAF treated with monotherapy and combination therapy in the CDX model using the cell line HT-29. V600E - MI and TGI values ​​of CRC

[0197] sample Concentration (uM) MI %TGI Osimertinib 0.5 1.20 16.9639％ Afatinib 0.2 1.24 19.2409％ Cobimetinib 0.4 331.41 99.6983％ trametinib 0.02 17.16 94.1714％ Palbociclib 0.15 4.64 78.4714％ Riboxini 5 11.24 91.1053％ Osimertinib + cobimetinib + palbociclib 0.5+0.4+0.15 23332.49 99.9957％ Osimertinib + cobimetinib + ribociclib 0.5+0.4+5 12790.35 99.9922％ Osimertinib + trametinib + palbociclib 0.5+0.02+0.15 24849.10 99.9960％ Osimertinib + trametinib + ribociclib 0.5+0.02+5 24849.10 99.9960％ Afatinib + cobimetinib + palbociclib 0.2+0.4+0.15 24849.10 99.9960％ Afatinib + cobimetinib + ribociclib 0.2+0.4+5 24849.10 99.9960％ Afatinib + trametinib + palbociclib 0.2+0.02+0.15 6598.97 99.9848％ Afatinib + trametinib + ribociclib 0.2+0.02+5 24849.10 99.9960％

[0198] Figure 8 and Figure 9Figure 2 shows the changes in tumor volume during treatment with various single agents and dual and triple combination therapies in the CDX model using the cell line HT-29. All four combinations tested (1) osimertinib + cobimetinib + palbociclib, (2) cetuximab + cobimetinib + palbociclib, (3) osimertinib + TAK-733 + palbociclib, and (4) cetuximab + TAK-733 + palbociclib reduced tumor volume by >50% in the first treatment cycle (21 days). Chemotherapeutic agents of capecitabine (squares) and 5-FU (dashed circles) showed no efficacy, and tumor growth trends were similar to those of the vehicle control (triangles). The triple combination of cetuximab, cobimetinib and palbociclib and the triple combination of osimertinib, cobimetinib and palbociclib showed superior tumor growth inhibition compared to monotherapy (cobimetinib) and the double combination of cobimetinib and osimertinib and the double combination of cobimetinib and palbociclib (see also Table 9).

[0199] Table 9. Tumor growth inhibition of the CDX model using the cell line HT-29

[0200]

[0201] Example 7. Clinical study of combination therapy

[0202] In patients with either BRAF V600E A Phase 2 open-label clinical study was conducted in patients with advanced colorectal cancer with BRAF mutations. The study was designed to evaluate the safety and efficacy of the combination therapy (cetuximab + cobimetinib + palbociclib) in patients with advanced colorectal cancer. The study subject population included adult men and women aged 18-75 years who were diagnosed with either BRAF V600E mutation in advanced colorectal cancer and with an expected survival of longer than 12 months.

[0203] Subjects were given: (1) cetuximab at a maximum infusion rate of 10 mg / min and a dose of 400 mg / m 2 Infuse over 120 minutes for 1 week, then 250 mg / m once weekly 2 (1) cobimetinib, 20-60 mg orally daily; and (2) palbociclib, 75-125 mg orally daily. Vital signs, clinical laboratory measurements, resting ECG measurements, adverse events, and serious adverse events were monitored and assessed. Overall response rate, disease control rate, overall survival, duration of response, and quality of life were also monitored and assessed. Study parameters are summarized in Table 10.

[0204] Table 10

[0205]

[0206]

Claims

1. Use of a combination of (a), (b) and (c) in the preparation of a medicament for treating cancer in a subject or delaying its progression: (a) Epidermal growth factor receptor (EGFR) inhibitors; (b) mitogen-activated protein kinase (MEK) 1 / 2 inhibitors; and (c) cyclin-dependent kinase (CDK) 4 / 6 inhibitors; wherein the EGFR inhibitor is cetuximab, the MEK 1 / 2 inhibitor is cobimetinib or a salt thereof, and the CDK 4 / 6 inhibitor is palbociclib or a salt thereof, wherein a BRAF inhibitor is not administered to the subject, and wherein the cancer is colorectal cancer having a BRAF V600E mutation.

2. The use according to claim 1, wherein cetuximab, cobimetinib or a salt thereof, and palbociclib or a salt thereof are administered in two or more compositions. 3 . The use according to claim 1 , wherein cetuximab, cobimetinib or a salt thereof, and palbociclib or a salt thereof are administered to the subject intermittently.

4. The use according to claim 1, wherein cobimetinib or a salt thereof and palbociclib or a salt thereof are administered in one composition.

5. The use according to claim 1, wherein cobimetinib or a salt thereof and palbociclib or a salt thereof are formulated for oral administration. The use according to claim 1 , wherein cobimetinib or a salt thereof and palbociclib or a salt thereof are administered to the subject simultaneously.

7. The use according to claim 1, wherein the cancer is stage IV colorectal cancer.

8. The use according to claim 1, wherein the treatment or delay results in a reduction in cancer cell growth and / or an increase in cancer cell killing that is 20-99% greater than that of cetuximab, cobimetinib or a salt thereof, or palbociclib or a salt thereof administered alone.

9. The use according to claim 1, wherein the treatment or delay reduces the mean tumor volume by 20-95%. 10 . The use according to claim 1 , wherein cobimetinib or a salt thereof is administered to the subject at a daily dose of 0.25-1 mg / kg.

11. The use according to claim 1, wherein palbociclib or a salt thereof is administered to the subject at a daily dose of 1-2.5 mg / kg.

12. The method according to claim 1, wherein cetuximab is administered to the subject as follows: 400 mg / m2 infused over 120 minutes at a maximum infusion rate of 10 mg / min. 2 weekly dose, followed by 250 mg / m2 over 60 minutes at a maximum infusion rate of 10 mg / min 2 weekly dose.

13. The use according to claim 1, wherein the subject is a human.

14. The use of claim 1, wherein the treating or delaying reduces tumor volume by at least 85%.

15. A kit comprising: (a) Epidermal growth factor receptor (EGFR) inhibitors; (b) mitogen-activated protein kinase (MEK) 1 / 2 inhibitors; and (c) cyclin-dependent kinase (CDK) 4 / 6 inhibitors; wherein the EGFR inhibitor is cetuximab, the MEK 1 / 2 inhibitor is cobimetinib or a salt thereof, and the CDK 4 / 6 inhibitor is palbociclib or a salt thereof, and wherein the kit does not comprise a BRAF inhibitor.

16. The kit according to claim 15, wherein the kit comprises a pharmaceutically acceptable carrier.

17. The kit according to claim 16, wherein the pharmaceutically acceptable carrier is an excipient, a binder or a diluent.

18. The kit of claim 15, wherein cobimetinib or a salt thereof is formulated for oral administration to a subject.

19. The kit of claim 15, wherein palbociclib or a salt thereof is formulated for oral administration to a subject.

20. The kit of claim 15, wherein the cetuximab is formulated for oral administration to a subject.

21. The kit according to claim 15, wherein cetuximab, cobimetinib or a salt thereof, and palbociclib or a salt thereof are formulated in two or more compositions.

22. The kit of claim 15, wherein cobimetinib or a salt thereof and palbociclib or a salt thereof are administered in one composition.

23. The kit according to claim 15, wherein cobimetinib or a salt thereof and palbociclib or a salt thereof are administered to the subject simultaneously.

24. A combination drug comprising: (a) Epidermal growth factor receptor (EGFR) inhibitors; (b) mitogen-activated protein kinase (MEK) 1 / 2 inhibitors; and (c) cyclin-dependent kinase (CDK) 4 / 6 inhibitors; wherein the EGFR inhibitor is cetuximab, the MEK 1 / 2 inhibitor is cobimetinib or a salt thereof, and the CDK 4 / 6 inhibitor is palbociclib or a salt thereof, wherein the combination drug does not contain a BRAF inhibitor; and wherein (a), (b) and (c) are formulated separately and used simultaneously or separately. The combination drug according to claim 24 , further comprising a pharmaceutically acceptable carrier.

26. The combination drug according to claim 25, wherein the pharmaceutically acceptable carrier is an excipient, a binder or a diluent.

27. The combination pharmaceutical according to claim 24 or 25, wherein the combination pharmaceutical is formulated for oral administration to a subject.

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