Treatment of Cancer Using Ketotifen in Combination with Checkpoint Inhibitors

Abstract

The present invention is directed to a combination treatment using ketotifen and checkpoint inhibitors, which is effective for treating cancer or inhibiting the growth of tumor cells in a subject, and / or can induce, enhance or prolong immune response against tumor cells.The effectiveness of cancer immunotherapy depends on whether T cells can migrate into tumors and migrate to adjacent sites of malignant cells to recognize and kill them.The present invention provides a means to solve this problem.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 287,872, filed December 9, 2021, the contents of which are incorporated herein by reference.

[0002] INCORPORATION BY REFERENCE TO SEQUENCE LISTING The contents of the electronic sequence listing (211482000240SEQLIST.xml; size: 23,937 bytes; and creation date: December 8, 2022) are incorporated herein by reference in their entirety.

[0003] FIELD OF THEINVENTION The present invention discloses combination treatments using ketotifen and checkpoint inhibitors that are effective for treating cancer or inhibiting the growth of tumor cells in a subject and / or can elicit, enhance or prolong an immune response against tumor cells. [Background technology]

[0004] background The efficacy of cancer immunotherapy depends in part on whether T cells can migrate into tumors and migrate adjacent to malignant cells to recognize and kill them. One barrier to T cell homing is the tumor vascular wall, which inhibits T cell attachment and migration through the endothelin B receptor, but antagonizing this receptor has not yet led to a clinically approved drug. One reason may be hypoperfusion in tumors, which may limit the surface area of ​​perfused blood vessels for antitumor T cells to attach. If the collapsed tumor vessels could be decompressed and reperfused by relieving mechanical compression (i.e., solid stress), antagonizing the endothelin B receptor could increase the efficacy of cancer immunotherapy.

[0005] Ketotifen is a cycloheptathiophene derivative drug that was first marketed as an inhibitor of anaphylaxis. Ketotifen selectively blocks histamine (H1) receptors, suppressing histamine release and associated symptoms. Ketotifen has also been reported to inhibit mast cell activity.

[0006] Immune checkpoints, which act as off switches in the immune system's T cells, are being investigated to restore immune responses with targeted agents, thus indirectly treating cancer by activating the body's immune system.

[0007] International patent applications WO2002086083, WO2004004771, WO2004056875, WO2006121168, WO2008156712, WO2010077634, WO2011066389, WO2014055897, and WO2014100079 report PD-1, PD-L1 inhibitory antibodies and / or methods for identifying such antibodies. Additionally, U.S. patents such as U.S. Patent No. 8,735,553 and U.S. Patent No. 8,168,757 report PD-1 or PD-L1 inhibitory antibodies and / or fusion proteins. The disclosures of WO2002086083, WO2004004771, WO2004056875, WO2006121168, WO2008156712, WO2010077634, WO2011066389, WO2014055897, and WO2014100079, as well as U.S. Pat. No. 8,735,553 and U.S. Pat. No. 8,168,757, are incorporated by reference herein in their entireties.

[0008] Moreover, International Patent Applications WO2011161699, WO2012168944, WO2013144704, WO2013132317, and WO2016044900 report peptide or peptidomimetic compounds capable of suppressing and / or inhibiting the programmed cell death 1 (PD-1) signaling pathway. The disclosures of WO2011161699, WO2012168944, WO2013144704, WO2013132317, and WO2016044900 are incorporated herein by reference in their entireties.

[0009] Furthermore, International Patent Applications WO2016142852, WO2016142894, WO2016142886, WO2016142835, and WO2016142833 report small molecule compounds capable of suppressing and / or inhibiting the programmed cell death 1 (PD-1) signaling pathway and / or treating disorders by inhibiting immunosuppressive signals induced by PD-1, PD-L1, or PD-L2. The disclosures of WO2016142852, WO2016142894, WO2016142886, WO2016142835, and WO2016142833 are incorporated herein by reference in their entirety.

[0010] Recently, ipilimumab (Yervoy®), a monoclonal antibody that targets cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4), and nivolumab (Opdivo®), a monoclonal antibody that targets programmed cell death protein 1 pathway (PD-1) on the surface of T cells, have been approved by the U.S. Food and Drug Administration for the treatment of advanced melanoma, advanced renal cell carcinoma, and non-small cell lung cancer. However, current checkpoint inhibitor therapy is effective in treating cancer in a relatively small population of cancer subjects, in part due to the presence of pre-existing immune activation and inhibitory receptors. Immune checkpoint blockade (ICB) with checkpoint inhibitors has revolutionized the treatment of many types of solid tumors, but is currently estimated to benefit less than 20% of cancer patients. Increasing the proportion of patients who respond and the duration of that response is an urgent unmet clinical need. Therefore, there is a need to develop methods and combination therapies to trigger or enhance the effectiveness of checkpoint inhibitors in both non-responsive and responsive subject populations.

[0011] Before antitumor T cells can encounter cancer cells, they must circulate through tumors via blood vessels, bind to the endothelium, cross the blood vessel wall, and migrate through cancer-associated fibroblasts (CAFs) and extracellular matrix (ECM). However, up to 80% of intratumoral blood vessels lack perfusion, limiting the area of ​​the blood vessel wall through which T cells can migrate.

[0012] Compressed blood vessels impair blood flow and oxygen delivery to tumors, leading to increased hypoxia in tumors and resistance to immunotherapy through multiple mechanisms. Vascular decompression strategies enhance the efficacy of ICB in an ICB-resistant mouse metastatic breast cancer model. If there was a way to decompress tumor vasculature while also facilitating T cell adhesion and migration to the tumor parenchyma, the proportion of cancer patients who respond to ICB would increase. All references cited herein, including patent applications, patent publications, and scientific articles, are incorporated by reference in their entirety herein, as if each individual reference was specifically and individually indicated to be incorporated by reference herein. [Prior art documents] [Patent documents]

[0013] [Patent Document 1] International Publication No. 2002 / 086083 [Patent Document 2] International Publication No. 2004 / 004771 [Patent Document 3] International Publication No. 2004 / 056875 [Patent Document 4] International Publication No. 2006 / 121168 [Patent Document 5] International Publication No. 2008 / 156712 [Patent Document 6] International Publication No. 2010 / 077634 [Patent Document 7] International Publication No. 2011 / 066389 [Patent Document 8] International Publication No. 2014 / 055897 [Patent Document 9] International Publication No. 2014 / 100079 [Patent Document 10] U.S. Patent No. 8,735,553 [Patent Document 11] U.S. Patent No. 8,168,757 [Patent Document 12] International Publication No. 2011 / 161699 [Patent Document 13] International Publication No. 2012 / 168944 [Patent Document 14] International Publication No. 2013 / 144704 [Patent Document 15] International Publication No. 2013 / 132317 [Patent Document 16] International Publication No. 2016 / 044900 [Patent Document 17] International Publication No. 2016 / 142852 [Patent Document 18] International Publication No. 2016 / 142894 [Patent Document 19] International Publication No. 2016 / 142886 [Patent Document 20] International Publication No. 2016 / 142835 [Patent Document 21] International Publication No. 2016 / 142833 Summary of the Invention [Means for solving the problem]

[0014] Abstract Provided herein is a method for treating a solid tumor in a subject in need thereof, comprising administering to the subject an effective amount of ketotifen or a pharma- ceutically acceptable salt thereof in combination with a checkpoint inhibitor. Also provided herein is a method for initiating, enhancing, or prolonging the effect of a checkpoint inhibitor in a subject in need thereof, or enabling the subject to respond to a checkpoint inhibitor, comprising administering to the subject an effective amount of ketotifen or a pharma- ceutically acceptable salt thereof in combination with a checkpoint inhibitor, wherein the subject has a solid tumor. Also provided herein is a method for enhancing the effect of a checkpoint inhibitor in a subject in need thereof, comprising administering to the subject an effective amount of ketotifen or a pharma- ceutically acceptable salt thereof in combination with a checkpoint inhibitor, wherein the subject has a solid tumor. Also provided herein is a method for increasing blood flow of a solid tumor in a subject, comprising administering to the subject an effective amount of ketotifen or a pharma- ceutically acceptable salt thereof in combination with a checkpoint inhibitor, wherein increasing blood flow of the solid tumor enhances the effect of the checkpoint inhibitor. In some embodiments, blood flow is measured using ultrasound-based blood flow measurements or using histology techniques to measure hypoxia. Also provided herein is a method of improving delivery or efficacy of a checkpoint inhibitor in a subject, comprising administering an effective amount of ketotifen or a pharmaceutically acceptable salt thereof in combination with a checkpoint inhibitor, the subject having a solid tumor, thereby improving delivery or efficacy of the treatment in the subject. In some embodiments, administering ketotifen or a pharmaceutically acceptable salt thereof increases the number of anti-tumor T cells co-localized with the solid tumor. In some embodiments, administering ketotifen or a pharmaceutically acceptable salt thereof. Reduces tissue stiffness of the solid tumor. In some embodiments, tissue stiffness of the solid tumor is measured using ultrasound elastography. In some embodiments, administering ketotifen or a pharmaceutically acceptable salt thereof reduces the level of extracellular matrix proteins in the solid tumor.In some embodiments, the extracellular matrix protein is collagen I or hyaluronan binding protein (HABP). In some embodiments, administering ketotifen or a pharma- ceutically acceptable salt thereof reduces hypoxia in solid tumors. In some embodiments, the checkpoint inhibitor inhibits a checkpoint protein selected from the group consisting of CTLA-4, PD-1, PD-L1, PD-L2, B7-H3, B7-H4, BMA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, and B-7 family ligands or combinations thereof. In some embodiments, the checkpoint inhibitor is a CTLA-4, PD-L1, PD-L2, or PD-1 inhibitor. In some embodiments, the checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA4 antibody. In some embodiments, the checkpoint inhibitor is selected from the group consisting of MEDI0680, AMP-224, nivolumab, pembrolizumab, pidilizumab, MEDI4736, atezolizumab, ipilimumab, tremelimumab, and BMS-936559. In some embodiments, the checkpoint inhibitor is a combination of an anti-PD-1 antibody and an anti-CTLA-4 antibody. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof is administered to the subject once a day. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof is administered to the subject twice a day. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof is administered to the subject at a dose of about 0.01 mg / kg to about 5 mg / kg. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof is administered to the subject at a dose of about 100 mg to about 1200 mg. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof is administered to the subject at a dose of about 125 mg to about 500 mg. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof is administered to the subject at a dose of about 125 mg. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof is administered to the subject at a dose of about 500 mg. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof is administered to the subject prior to administering a checkpoint inhibitor to the subject. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof is administered to the subject beginning at least one day prior to administering the checkpoint inhibitor to the subject. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof is administered to the subject beginning at least two days prior to administering the checkpoint inhibitor to the subject. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof is administered to the subject beginning at least three days prior to administering the checkpoint inhibitor to the subject. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof is administered to the subject beginning at least five days prior to administering the checkpoint inhibitor to the subject. In some embodiments, administration of ketotifen or a pharma- ceutically acceptable salt thereof to the subject is maintained for at least a portion of the period during which the checkpoint inhibitor is administered to the subject. In some embodiments, administration of ketotifen or a pharma- ceutically acceptable salt thereof to the subject is maintained for the entire period during which the checkpoint inhibitor is administered to the subject. In some embodiments, one or more therapeutic effects in the subject improves compared to baseline after administration of ketotifen or a pharma- ceutically acceptable salt thereof and the checkpoint inhibitor.In some embodiments, the one or more therapeutic effects are selected from the group consisting of the size of the tumor originating from the cancer, objective response rate, duration of response, time to response, progression-free survival, and overall survival.In some embodiments, the size of the tumor originating from the cancer is reduced by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, or at least about 80% compared to the size of the tumor originating from the cancer before administration of ketotifen or its pharma- ceutically acceptable salt and checkpoint inhibitor.In some embodiments, the objective response rate is at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, or at least about 80%. In some embodiments, the subject exhibits progression-free survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration of ketotifen or a pharmaceutically acceptable salt thereof and a checkpoint inhibitor. In some embodiments, the subject exhibits an overall survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration of ketotifen or a pharmaceutically acceptable salt thereof and a checkpoint inhibitor.In some embodiments, the duration of response to the antibody-drug conjugate is at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof and a checkpoint inhibitor. In some embodiments, the solid tumor is selected from the group consisting of mesothelioma, breast cancer, lung metastasis of breast cancer, sarcoma, pancreatic cancer, ovarian cancer, liver metastasis, prostate cancer, brain cancer, melanoma, renal cell carcinoma, colorectal cancer, hepatocellular carcinoma, lung cancer, head and neck squamous cell carcinoma, urothelial carcinoma, esophageal squamous cell carcinoma, gastric cancer, esophageal cancer, cervical cancer, Merkel cell carcinoma, endometrial carcinoma, and cutaneous squamous cell carcinoma. In some embodiments, the solid tumor is a sarcoma. In some embodiments, the sarcoma is osteosarcoma or fibrosarcoma. In some embodiments, the subject is a human. In some embodiments of any of the above methods, the method further comprises administering an additional chemotherapeutic agent. In some embodiments, the additional chemotherapeutic agent is doxorubicin or an analog or derivative thereof.

[0015] Also provided herein are kits comprising: (a) an effective amount of ketotifen or a pharma- ceutically acceptable salt thereof; (b) an effective amount of a checkpoint inhibitor; and (c) instructions for using ketotifen or a pharma-ceutically acceptable salt thereof and a checkpoint inhibitor in accordance with any of the methods described herein.

[0016] Also provided herein is a method for determining an effective amount of ketotifen in a subject having a solid tumor, comprising: (a) measuring the blood flow and / or hardness of the solid tumor; (b) administering an effective amount of ketotifen to the subject; and (c) measuring the blood flow and / or hardness of the solid tumor after administration of ketotifen, where an increase in blood flow and / or a decrease in hardness after administration of ketotifen to the subject indicates that the amount administered was an effective amount. Also provided herein is a method for treating a solid tumor in a subject in need thereof, comprising: (a) measuring the blood flow and / or hardness of the solid tumor; (b) administering an effective amount of ketotifen to the subject; (c) measuring the blood flow and / or hardness of the solid tumor after administration of ketotifen; and (d) administering a chemotherapeutic agent if the blood flow of the solid tumor increases and / or the hardness of the solid tumor decreases after administration of ketotifen. Also provided herein is a method of treating a solid tumor in a subject in need thereof, comprising: (a) measuring the blood flow and / or hardness of the solid tumor; (b) administering an effective amount of ketotifen to the subject; (c) measuring the blood flow and / or hardness of the solid tumor after administration of ketotifen; (d) determining that the subject is responsive to the chemotherapeutic agent based on an increase in blood flow of the solid tumor or a decrease in hardness of the solid tumor after administration of ketotifen; and (e) administering the chemotherapeutic agent to the subject who has been determined to be responsive to the chemotherapeutic agent based on an increase in blood flow of the solid tumor or a decrease in hardness of the solid tumor after administration of ketotifen. Also provided herein is a method for predicting response to treatment with a chemotherapeutic agent, comprising: (a) measuring the blood flow and / or hardness of a solid tumor; (b) administering to a subject an effective amount of ketotifen; and (c) measuring the blood flow and / or hardness of the solid tumor after administration of ketotifen, wherein an increase in blood flow of the solid tumor or a decrease in hardness of the solid tumor after administration of ketotifen indicates that the subject is likely to respond to treatment with the chemotherapeutic agent.In some embodiments, the effective amount of ketotifen is determined by measuring a change in blood flow and / or stiffness of a solid tumor following administration of ketotifen to the subject, where an increase in blood flow and / or a decrease in stiffness following administration of ketotifen to the subject indicates that the amount administered was an effective amount. In some embodiments, the method includes measuring blood flow of the solid tumor, where blood flow of the solid tumor increases following administration of ketotifen. In some embodiments, the method includes measuring stiffness of the solid tumor, where stiffness of the solid tumor decreases following administration of ketotifen. In some embodiments, ketotifen is administered at least 1 day, at least 2 days, at least 3 days, at least 4 days, or at least 5 days prior to administration of the chemotherapeutic agent. In some embodiments, ketotifen is administered at a dose that increases blood flow of the solid tumor and / or decreases stiffness of the solid tumor. In some embodiments, blood flow and / or stiffness of the solid tumor is measured using ultrasound. In some embodiments, blood flow of the solid tumor is measured using a histology technique to measure hypoxia. In some embodiments, the chemotherapeutic agent is a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor inhibits a checkpoint protein selected from the group consisting of CTLA-4, PD-1, PD-L1, PD-L2, B7-H3, B7-H4, BMA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, and B-7 family ligands or combinations thereof. In some embodiments, the checkpoint inhibitor is a CTLA-4, PD-L1, PD-L2, or PD-1 inhibitor. In some embodiments, the checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA4 antibody. In some embodiments, the checkpoint inhibitor is selected from the group consisting of MEDI0680, AMP-224, nivolumab, pembrolizumab, pidilizumab, MEDI4736, atezolizumab, ipilimumab, tremelimumab, and BMS-936559. In some embodiments, the checkpoint inhibitor is a combination of an anti-PD-1 antibody and an anti-CTLA-4 antibody.In some embodiments, the solid tumor is selected from the group consisting of breast cancer, lung metastasis of breast cancer, sarcoma, pancreatic cancer, ovarian cancer, liver metastasis, prostate cancer, brain cancer, melanoma, renal cell carcinoma, colorectal cancer, hepatocellular carcinoma, lung cancer, head and neck squamous cell carcinoma, urothelial carcinoma, esophageal squamous cell carcinoma, gastric cancer, esophageal cancer, cervical cancer, Merkel cell carcinoma, endometrial carcinoma, and cutaneous squamous cell carcinoma. In some embodiments, the solid tumor is a sarcoma. In some embodiments, the sarcoma is osteosarcoma or fibrosarcoma. In some embodiments, the subject is a human. In some embodiments of the method, the chemotherapeutic agent is doxorubicin or an analog or derivative thereof.

[0017] It should be understood that one, some, or all of the characteristics of the various embodiments described herein may be combined to form other embodiments of the present invention. These and other aspects of the present invention will be apparent to those skilled in the art. These and other embodiments of the present invention are further described by the following detailed description.

[0018] Exemplary embodiments of the present invention are disclosed by reference to the following drawings, in which it should be understood that the depicted embodiments are not limited to the precise details shown. [Brief description of the drawings]

[0019] [Figure 1A]Figure 1A shows representative immunofluorescence images of paraffin sections of MCA205 tumor models stained for pimonidazole adducts after pimonidazole hydrochloride injection; scale bar = 0.2 mm (top two rows are DAPI staining, bottom two rows are corresponding hypoxic staining). Figures 1B-1C show quantification of the fraction of hypoxic areas normalized to DAPI staining in MCA205 (Figure 1B) and K7M2wt (Figure 1C) tumors (n = 5 mice / group, N = 3-5 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. FIG. 1D shows quantification of IFN-γ and VEGF mRNA expression levels in untreated and ketotifen (10 mg / kg)-treated MCA205 tumors using the 2^-ΔΔCT method (3 biological replicates × 3 technical replicates were used). All data are presented as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups to control groups * and 10 mg / kg groups to all other treatment groups **. p≦0.05. FIG. 1E shows IFP levels in untreated mice and mice treated with ketotifen daily for 7 days (n=7 mice / treatment group). All data are presented as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups to control groups * and 10 mg / kg groups to all other treatment groups **. p≦0.05. [Figure 1B-C]Figure 1A shows representative immunofluorescence images of paraffin sections of MCA205 tumor models stained for pimonidazole adducts after pimonidazole hydrochloride injection; scale bar = 0.2 mm (top two rows are DAPI staining, bottom two rows are corresponding hypoxic staining). Figure 1B-1C show quantification of the percentage of hypoxic areas normalized to DAPI staining in MCA205 (Figure 1B) and K7M2wt (Figure 1C) tumors (n = 5 mice / group, N = 3-5 image fields per mouse). All data are expressed as the mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. FIG. 1D shows quantification of IFN-γ and VEGF mRNA expression levels in untreated and ketotifen (10 mg / kg)-treated MCA205 tumors using the 2^-ΔΔCT method (3 biological replicates × 3 technical replicates were used). All data are presented as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups to control groups * and 10 mg / kg groups to all other treatment groups **. p≦0.05. FIG. 1E shows IFP levels in untreated mice and mice treated with ketotifen daily for 7 days (n=7 mice / treatment group). All data are presented as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups to control groups * and 10 mg / kg groups to all other treatment groups **. p≦0.05. [Fig. 1D-E]Figure 1A shows representative immunofluorescence images of paraffin sections of MCA205 tumor models stained for pimonidazole adducts after pimonidazole hydrochloride injection; scale bar = 0.2 mm (top two rows are DAPI staining, bottom two rows are corresponding hypoxic staining). Figure 1B-1C show quantification of the percentage of hypoxic areas normalized to DAPI staining in MCA205 (Figure 1B) and K7M2wt (Figure 1C) tumors (n = 5 mice / group, N = 3-5 image fields per mouse). All data are expressed as the mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. FIG. 1D shows quantification of IFN-γ and VEGF mRNA expression levels in untreated and ketotifen (10 mg / kg)-treated MCA205 tumors using the 2^-ΔΔCT method (3 biological replicates × 3 technical replicates were used). All data are presented as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups to control groups * and 10 mg / kg groups to all other treatment groups **. p≦0.05. FIG. 1E shows IFP levels in untreated mice and mice treated with ketotifen daily for 7 days (n=7 mice / treatment group). All data are presented as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups to control groups * and 10 mg / kg groups to all other treatment groups **. p≦0.05.

[0020] [Figure 2A-B]Figure 2A shows the growth curves of MCA205 tumors after daily administration of various doses of ketotifen (1, 5, 10 and 25 mg / kg) compared to the control. Figure 2B shows the growth curves of K7M2wt tumors after daily administration of 10 mg / kg of ketotifen compared to the control. Figures 2C-2D show measurements of MCA205 (Figure 2C) and K7M2wt (Figure 2D) tumor burden after completion of the treatment protocol. All data are expressed as the mean ± standard error of the mean (n=5-7 mice / treatment group). Figure 2E shows an exemplary Western blot for CD117 protein with β-actin as a loading control (n=4 mice / group). [Figure 2C-D] Figure 2A shows the growth curves of MCA205 tumors after daily administration of various doses of ketotifen (1, 5, 10 and 25 mg / kg) compared to the control. Figure 2B shows the growth curves of K7M2wt tumors after daily administration of 10 mg / kg of ketotifen compared to the control. Figures 2C-2D show measurements of MCA205 (Figure 2C) and K7M2wt (Figure 2D) tumor burden after completion of the treatment protocol. All data are expressed as the mean ± standard error of the mean (n=5-7 mice / treatment group). Figure 2E shows an exemplary Western blot for CD117 protein with β-actin as a loading control (n=4 mice / group). [Figure 2E] Figure 2A shows the growth curves of MCA205 tumors after daily administration of various doses of ketotifen (1, 5, 10 and 25 mg / kg) compared to the control. Figure 2B shows the growth curves of K7M2wt tumors after daily administration of 10 mg / kg of ketotifen compared to the control. Figures 2C-2D show measurements of MCA205 (Figure 2C) and K7M2wt (Figure 2D) tumor burden after completion of the treatment protocol. All data are expressed as the mean ± standard error of the mean (n=5-7 mice / treatment group). Figure 2E shows an exemplary Western blot for CD117 protein with β-actin as a loading control (n=4 mice / group).

[0021] [Figure 3A-F]Figures 3A-3F show body weights, thymus weights and spleen weights of mice at the conclusion of the study of MCA205 (Figures 3A-3C) and K7M2wt (Figures 3D-3F) tumors treated with control or the indicated doses of ketotifen. All data are expressed as mean ± standard error of the mean (n = 5-7 mice / treatment group). Statistical analysis was performed by comparing treatment groups with control group* and 10 mg / kg group. p ≤ 0.05.

[0022] [Figure 4A-C] FIG. 4A shows an exemplary study treatment protocol for the murine MCA205 tumor model with daily ketotifen treatment at an exemplary dose of 10 mg / kg. FIG. 4B and FIG. 4C show longitudinal measurements of tissue-level macroscopic Young's modulus in MCA205 (FIG. 4B) and K7M2wt (FIG. 4C) tumors treated as indicated, as assessed using ultrasound elastography (n=4 mice / group, N=2 image fields per mouse for ultrasound measurements). FIG. 4D-4E show vascular perfusion in MCA205 tumors treated as indicated 3 days (FIG. 4D) and 7 days (FIG. 4E) after ketotifen treatment. All data are expressed as mean±standard error of the mean (n=4 mice / treatment group). Statistical analysis was performed by comparing treatment groups to the control group* and the 10 mg / kg group. p≦0.05. Figures 4F-4G show functional perfusion in K7M2wt tumors treated as indicated after 3 days (Figure 4F) and 7 days (Figure 4G) of treatment. All data are expressed as mean ± standard error of the mean (n = 4 mice / treatment group). Statistical analysis was performed by comparing treatment groups with control group* and 10 mg / kg group. p ≤ 0.05. [Fig. 4D-G]FIG. 4A shows an exemplary study treatment protocol for the murine MCA205 tumor model with daily ketotifen treatment at an exemplary dose of 10 mg / kg. FIG. 4B and FIG. 4C show longitudinal measurements of tissue-level macroscopic Young's modulus in MCA205 (FIG. 4B) and K7M2wt (FIG. 4C) tumors treated as indicated, as assessed using ultrasound elastography (n=4 mice / group, N=2 image fields per mouse for ultrasound measurements). FIG. 4D-4E show vascular perfusion in MCA205 tumors treated as indicated 3 days (FIG. 4D) and 7 days (FIG. 4E) after ketotifen treatment. All data are expressed as mean±standard error of the mean (n=4 mice / treatment group). Statistical analysis was performed by comparing treatment groups to the control group* and the 10 mg / kg group. p≦0.05. Figures 4F-4G show functional perfusion in K7M2wt tumors treated as indicated after 3 days (Figure 4F) and 7 days (Figure 4G) of treatment. All data are expressed as mean ± standard error of the mean (n = 4 mice / treatment group). Statistical analysis was performed by comparing treatment groups with control group* and 10 mg / kg group. p ≤ 0.05.

[0023] [Figure 5A-B]Figure 5A and Figure 5B show representative bright field images of picrosirius red staining in paraffin tumor sections of MCA205 (Figure 5A) and K7M2wt (Figure 5B) (n = 4 mice / group; N = 5-6 image fields per mouse). Figure 5C and Figure 5D show quantification of areas positive for picrosirius red staining in fibrosarcoma MCA205 tumors (Figure 5C) and osteosarcoma K7M2wt tumors (Figure 5D) normalized to the control group. All data are expressed as the mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. Figures 5E-5F show representative immunofluorescence images of Ki-67 proliferation marker and α-SMA in fibrosarcoma tumors after treatment with control or the indicated doses of ketotifen. Scale bar = 0.2 mm (Figure 5E; top two rows are α-SMA and bottom two rows are the corresponding Ki67 staining); also shown is quantification of cancer-associated fibroblasts (CAFs) positive for both α-SMA and Ki-67 markers, normalized to total α-SMA staining (Figure 5F), after treatment with control or the indicated doses of ketotifen. All data are expressed as mean ± standard error of the mean (for tissue analysis, n = 4 mice / group, N = 5-6 image fields per mouse). FIG. 5G shows quantification of mRNA expression levels of Col1A1, CTGF, ACTA2, HAS2, and HAS3 in untreated (control) MCA205 tumors and MCA205 tumors treated with ketotifen (10 mg / kg) using the 2^-ΔΔCT method (3 biological replicates × 3 technical replicates were used). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≦ 0.05. FIG. 5H-5I show quantification of the percentage of area positive for α-SMA staining (FIG. 5H) and the percentage of area positive for Ki-67 staining (FIG. 5I) in MCA205 tumors after treatment with control or the indicated doses of ketotifen.All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Figures 5J-5L show representative immunofluorescence images of MCA205 paraffin sections stained with anti-HABP1, scale bar = 0.2 mm, after treatment with control or the indicated doses of ketotifen, respectively (Figure 5J; top two rows are DAPI staining, bottom two rows are corresponding HABP1 staining), quantification of the percentage of area positive for HABP1 staining in MCA205 tumors (Figure 5K), and quantification of solid stress measured by the amount of length of tumor opening after cutting the tissue (n = 4 mice / group) (Figure 5L). All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p≦0.05. [Figure 5C-D]Figure 5A and Figure 5B show representative bright field images of picrosirius red staining in paraffin tumor sections of MCA205 (Figure 5A) and K7M2wt (Figure 5B) (n = 4 mice / group; N = 5-6 image fields per mouse). Figure 5C and Figure 5D show quantification of areas positive for picrosirius red staining in fibrosarcoma MCA205 tumors (Figure 5C) and osteosarcoma K7M2wt tumors (Figure 5D) normalized to the control group. All data are expressed as the mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. Figures 5E-5F show representative immunofluorescence images of Ki-67 proliferation marker and α-SMA in fibrosarcoma tumors after treatment with control or the indicated doses of ketotifen. Scale bar = 0.2 mm (Figure 5E; top two rows are α-SMA and bottom two rows are the corresponding Ki67 staining); also shown is quantification of cancer-associated fibroblasts (CAFs) positive for both α-SMA and Ki-67 markers, normalized to total α-SMA staining (Figure 5F), after treatment with control or the indicated doses of ketotifen. All data are expressed as mean ± standard error of the mean (for tissue analysis, n = 4 mice / group, N = 5-6 image fields per mouse). FIG. 5G shows quantification of mRNA expression levels of Col1A1, CTGF, ACTA2, HAS2, and HAS3 in untreated (control) MCA205 tumors and MCA205 tumors treated with ketotifen (10 mg / kg) using the 2^-ΔΔCT method (3 biological replicates × 3 technical replicates were used). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≦ 0.05. FIG. 5H-5I show quantification of the percentage of area positive for α-SMA staining (FIG. 5H) and the percentage of area positive for Ki-67 staining (FIG. 5I) in MCA205 tumors after treatment with control or the indicated doses of ketotifen.All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Figures 5J-5L show representative immunofluorescence images of MCA205 paraffin sections stained with anti-HABP1, scale bar = 0.2 mm, after treatment with control or the indicated doses of ketotifen, respectively (Figure 5J; top two rows are DAPI staining, bottom two rows are corresponding HABP1 staining), quantification of the percentage of area positive for HABP1 staining in MCA205 tumors (Figure 5K), and quantification of solid stress measured by the amount of length of tumor opening after cutting the tissue (n = 4 mice / group) (Figure 5L). All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p≦0.05. [Figure 5E]Figure 5A and Figure 5B show representative bright field images of picrosirius red staining in paraffin tumor sections of MCA205 (Figure 5A) and K7M2wt (Figure 5B) (n = 4 mice / group; N = 5-6 image fields per mouse). Figure 5C and Figure 5D show quantification of areas positive for picrosirius red staining in fibrosarcoma MCA205 tumors (Figure 5C) and osteosarcoma K7M2wt tumors (Figure 5D) normalized to the control group. All data are expressed as the mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. Figures 5E-5F show representative immunofluorescence images of Ki-67 proliferation marker and α-SMA in fibrosarcoma tumors after treatment with control or the indicated doses of ketotifen. Scale bar = 0.2 mm (Figure 5E; top two rows are α-SMA and bottom two rows are the corresponding Ki67 staining); also shown is quantification of cancer-associated fibroblasts (CAFs) positive for both α-SMA and Ki-67 markers, normalized to total α-SMA staining (Figure 5F), after treatment with control or the indicated doses of ketotifen. All data are expressed as mean ± standard error of the mean (for tissue analysis, n = 4 mice / group, N = 5-6 image fields per mouse). FIG. 5G shows quantification of mRNA expression levels of Col1A1, CTGF, ACTA2, HAS2, and HAS3 in untreated (control) MCA205 tumors and MCA205 tumors treated with ketotifen (10 mg / kg) using the 2^-ΔΔCT method (3 biological replicates × 3 technical replicates were used). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≦ 0.05. FIG. 5H-5I show quantification of the percentage of area positive for α-SMA staining (FIG. 5H) and the percentage of area positive for Ki-67 staining (FIG. 5I) in MCA205 tumors after treatment with control or the indicated doses of ketotifen.All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Figures 5J-5L show representative immunofluorescence images of MCA205 paraffin sections stained with anti-HABP1, scale bar = 0.2 mm, after treatment with control or the indicated doses of ketotifen, respectively (Figure 5J; top two rows are DAPI staining, bottom two rows are corresponding HABP1 staining), quantification of the percentage of area positive for HABP1 staining in MCA205 tumors (Figure 5K), and quantification of solid stress measured by the amount of length of tumor opening after cutting the tissue (n = 4 mice / group) (Figure 5L). All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p≦0.05. [Fig. 5F-G]Figure 5A and Figure 5B show representative bright field images of picrosirius red staining in paraffin tumor sections of MCA205 (Figure 5A) and K7M2wt (Figure 5B) (n = 4 mice / group; N = 5-6 image fields per mouse). Figure 5C and Figure 5D show quantification of areas positive for picrosirius red staining in fibrosarcoma MCA205 tumors (Figure 5C) and osteosarcoma K7M2wt tumors (Figure 5D) normalized to the control group. All data are expressed as the mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. Figures 5E-5F show representative immunofluorescence images of Ki-67 proliferation marker and α-SMA in fibrosarcoma tumors after treatment with control or the indicated doses of ketotifen. Scale bar = 0.2 mm (Figure 5E; top two rows are α-SMA and bottom two rows are the corresponding Ki67 staining); also shown is quantification of cancer-associated fibroblasts (CAFs) positive for both α-SMA and Ki-67 markers, normalized to total α-SMA staining (Figure 5F), after treatment with control or the indicated doses of ketotifen. All data are expressed as mean ± standard error of the mean (for tissue analysis, n = 4 mice / group, N = 5-6 image fields per mouse). FIG. 5G shows quantification of mRNA expression levels of Col1A1, CTGF, ACTA2, HAS2, and HAS3 in untreated (control) MCA205 tumors and MCA205 tumors treated with ketotifen (10 mg / kg) using the 2^-ΔΔCT method (3 biological replicates × 3 technical replicates were used). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≦ 0.05. FIG. 5H-5I show quantification of the percentage of area positive for α-SMA staining (FIG. 5H) and the percentage of area positive for Ki-67 staining (FIG. 5I) in MCA205 tumors after treatment with control or the indicated doses of ketotifen.All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Figures 5J-5L show representative immunofluorescence images of MCA205 paraffin sections stained with anti-HABP1, scale bar = 0.2 mm, after treatment with control or the indicated doses of ketotifen, respectively (Figure 5J; top two rows are DAPI staining, bottom two rows are corresponding HABP1 staining), quantification of the percentage of area positive for HABP1 staining in MCA205 tumors (Figure 5K), and quantification of solid stress measured by the amount of length of tumor opening after cutting the tissue (n = 4 mice / group) (Figure 5L). All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p≦0.05. [Figure 5H-I]Figure 5A and Figure 5B show representative bright field images of picrosirius red staining in paraffin tumor sections of MCA205 (Figure 5A) and K7M2wt (Figure 5B) (n = 4 mice / group; N = 5-6 image fields per mouse). Figure 5C and Figure 5D show quantification of areas positive for picrosirius red staining in fibrosarcoma MCA205 tumors (Figure 5C) and osteosarcoma K7M2wt tumors (Figure 5D) normalized to the control group. All data are expressed as the mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. Figures 5E-5F show representative immunofluorescence images of Ki-67 proliferation marker and α-SMA in fibrosarcoma tumors after treatment with control or the indicated doses of ketotifen. Scale bar = 0.2 mm (Figure 5E; top two rows are α-SMA and bottom two rows are the corresponding Ki67 staining); also shown is quantification of cancer-associated fibroblasts (CAFs) positive for both α-SMA and Ki-67 markers, normalized to total α-SMA staining (Figure 5F), after treatment with control or the indicated doses of ketotifen. All data are expressed as mean ± standard error of the mean (for tissue analysis, n = 4 mice / group, N = 5-6 image fields per mouse). FIG. 5G shows quantification of mRNA expression levels of Col1A1, CTGF, ACTA2, HAS2, and HAS3 in untreated (control) MCA205 tumors and MCA205 tumors treated with ketotifen (10 mg / kg) using the 2^-ΔΔCT method (3 biological replicates × 3 technical replicates were used). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≦ 0.05. FIG. 5H-5I show quantification of the percentage of area positive for α-SMA staining (FIG. 5H) and the percentage of area positive for Ki-67 staining (FIG. 5I) in MCA205 tumors after treatment with control or the indicated doses of ketotifen.All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Figures 5J-5L show representative immunofluorescence images of MCA205 paraffin sections stained with anti-HABP1, scale bar = 0.2 mm, after treatment with control or the indicated doses of ketotifen, respectively (Figure 5J; top two rows are DAPI staining, bottom two rows are corresponding HABP1 staining), quantification of the percentage of area positive for HABP1 staining in MCA205 tumors (Figure 5K), and quantification of solid stress measured by the amount of length of tumor opening after cutting the tissue (n = 4 mice / group) (Figure 5L). All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p≦0.05. [Figure 5J]Figure 5A and Figure 5B show representative bright field images of picrosirius red staining in paraffin tumor sections of MCA205 (Figure 5A) and K7M2wt (Figure 5B) (n = 4 mice / group; N = 5-6 image fields per mouse). Figure 5C and Figure 5D show quantification of areas positive for picrosirius red staining in fibrosarcoma MCA205 tumors (Figure 5C) and osteosarcoma K7M2wt tumors (Figure 5D) normalized to the control group. All data are expressed as the mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. Figures 5E-5F show representative immunofluorescence images of Ki-67 proliferation marker and α-SMA in fibrosarcoma tumors after treatment with control or the indicated doses of ketotifen. Scale bar = 0.2 mm (Figure 5E; top two rows are α-SMA and bottom two rows are the corresponding Ki67 staining); also shown is quantification of cancer-associated fibroblasts (CAFs) positive for both α-SMA and Ki-67 markers, normalized to total α-SMA staining (Figure 5F), after treatment with control or the indicated doses of ketotifen. All data are expressed as mean ± standard error of the mean (for tissue analysis, n = 4 mice / group, N = 5-6 image fields per mouse). FIG. 5G shows quantification of mRNA expression levels of Col1A1, CTGF, ACTA2, HAS2, and HAS3 in untreated (control) MCA205 tumors and MCA205 tumors treated with ketotifen (10 mg / kg) using the 2^-ΔΔCT method (3 biological replicates × 3 technical replicates were used). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≦ 0.05. FIG. 5H-5I show quantification of the percentage of area positive for α-SMA staining (FIG. 5H) and the percentage of area positive for Ki-67 staining (FIG. 5I) in MCA205 tumors after treatment with control or the indicated doses of ketotifen.All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Figures 5J-5L show representative immunofluorescence images of MCA205 paraffin sections stained with anti-HABP1, scale bar = 0.2 mm, after treatment with control or the indicated doses of ketotifen, respectively (Figure 5J; top two rows are DAPI staining, bottom two rows are corresponding HABP1 staining), quantification of the percentage of area positive for HABP1 staining in MCA205 tumors (Figure 5K), and quantification of solid stress measured by the amount of length of tumor opening after cutting the tissue (n = 4 mice / group) (Figure 5L). All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p≦0.05. [Figure 5K-L]Figure 5A and Figure 5B show representative bright field images of picrosirius red staining in paraffin tumor sections of MCA205 (Figure 5A) and K7M2wt (Figure 5B) (n = 4 mice / group; N = 5-6 image fields per mouse). Figure 5C and Figure 5D show quantification of areas positive for picrosirius red staining in fibrosarcoma MCA205 tumors (Figure 5C) and osteosarcoma K7M2wt tumors (Figure 5D) normalized to the control group. All data are expressed as the mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. Figures 5E-5F show representative immunofluorescence images of Ki-67 proliferation marker and α-SMA in fibrosarcoma tumors after treatment with control or the indicated doses of ketotifen. Scale bar = 0.2 mm (Figure 5E; top two rows are α-SMA and bottom two rows are the corresponding Ki67 staining); also shown is quantification of cancer-associated fibroblasts (CAFs) positive for both α-SMA and Ki-67 markers, normalized to total α-SMA staining (Figure 5F), after treatment with control or the indicated doses of ketotifen. All data are expressed as mean ± standard error of the mean (for tissue analysis, n = 4 mice / group, N = 5-6 image fields per mouse). FIG. 5G shows quantification of mRNA expression levels of Col1A1, CTGF, ACTA2, HAS2, and HAS3 in untreated (control) MCA205 tumors and MCA205 tumors treated with ketotifen (10 mg / kg) using the 2^-ΔΔCT method (3 biological replicates × 3 technical replicates were used). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≦ 0.05. FIG. 5H-5I show quantification of the percentage of area positive for α-SMA staining (FIG. 5H) and the percentage of area positive for Ki-67 staining (FIG. 5I) in MCA205 tumors after treatment with control or the indicated doses of ketotifen.All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Figures 5J-5L show representative immunofluorescence images of MCA205 paraffin sections stained with anti-HABP1, scale bar = 0.2 mm, after treatment with control or the indicated doses of ketotifen, respectively (Figure 5J; top two rows are DAPI staining, bottom two rows are corresponding HABP1 staining), quantification of the percentage of area positive for HABP1 staining in MCA205 tumors (Figure 5K), and quantification of solid stress measured by the amount of length of tumor opening after cutting the tissue (n = 4 mice / group) (Figure 5L). All data are expressed as mean ± standard error of the mean (for histological analysis, n = 4 mice / group, N = 5-6 image fields per mouse). Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p≦0.05.

[0024] [Figure 6A]Figure 6A shows representative immunofluorescence images of Ki-67 proliferation marker (bottom row) and α-SMA (top row; top and bottom rows show the same field) in K7M2wt tumors after treatment with control or the indicated doses of ketotifen. Scale bar = 0.2 mm. Figure 6B shows quantification of cancer-associated fibroblasts (CAFs) positive for both α-SMA and Ki-67 markers, normalized to total α-SMA staining, after treatment with control or the indicated doses of ketotifen (n = 4 mice / group, N = 5-6 image fields per mouse). Figure 6C shows quantification of the percentage of area positive for α-SMA staining after treatment with ketotifen compared to control. Figure 6D shows quantification of the percentage of area positive for Ki-67 staining in K7M2wt tumors after treatment with control or the indicated doses of ketotifen. Figure 6E shows representative immunofluorescence images of K7M2wt paraffin sections stained with anti-HABP1 (top row is DAPI staining, bottom row is corresponding HABP1 staining). Scale bar = 0.2 mm. Figure 6F shows quantification of the percentage of areas positive for HABP1 staining (n = 4 mice / group, N = 5-6 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. [Figure 6B-D]Figure 6A shows representative immunofluorescence images of Ki-67 proliferation marker (bottom row) and α-SMA (top row; top and bottom rows show the same field) in K7M2wt tumors after treatment with control or the indicated doses of ketotifen. Scale bar = 0.2 mm. Figure 6B shows quantification of cancer-associated fibroblasts (CAFs) positive for both α-SMA and Ki-67 markers, normalized to total α-SMA staining, after treatment with control or the indicated doses of ketotifen (n = 4 mice / group, N = 5-6 image fields per mouse). Figure 6C shows quantification of the percentage of area positive for α-SMA staining after treatment with ketotifen compared to control. Figure 6D shows quantification of the percentage of area positive for Ki-67 staining in K7M2wt tumors after treatment with control or the indicated doses of ketotifen. Figure 6E shows representative immunofluorescence images of K7M2wt paraffin sections stained with anti-HABP1 (top row is DAPI staining, bottom row is corresponding HABP1 staining). Scale bar = 0.2 mm. Figure 6F shows quantification of the percentage of areas positive for HABP1 staining (n = 4 mice / group, N = 5-6 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. [Figure 6E]Figure 6A shows representative immunofluorescence images of Ki-67 proliferation marker (bottom row) and α-SMA (top row; top and bottom rows show the same field) in K7M2wt tumors after treatment with control or the indicated doses of ketotifen. Scale bar = 0.2 mm. Figure 6B shows quantification of cancer-associated fibroblasts (CAFs) positive for both α-SMA and Ki-67 markers, normalized to total α-SMA staining, after treatment with control or the indicated doses of ketotifen (n = 4 mice / group, N = 5-6 image fields per mouse). Figure 6C shows quantification of the percentage of area positive for α-SMA staining after treatment with ketotifen compared to control. Figure 6D shows quantification of the percentage of area positive for Ki-67 staining in K7M2wt tumors after treatment with control or the indicated doses of ketotifen. Figure 6E shows representative immunofluorescence images of K7M2wt paraffin sections stained with anti-HABP1 (top row is DAPI staining, bottom row is corresponding HABP1 staining). Scale bar = 0.2 mm. Figure 6F shows quantification of the percentage of areas positive for HABP1 staining (n = 4 mice / group, N = 5-6 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05. [Figure 6F]Figure 6A shows representative immunofluorescence images of Ki-67 proliferation marker (bottom row) and α-SMA (top row; top and bottom rows show the same field) in K7M2wt tumors after treatment with control or the indicated doses of ketotifen. Scale bar = 0.2 mm. Figure 6B shows quantification of cancer-associated fibroblasts (CAFs) positive for both α-SMA and Ki-67 markers, normalized to total α-SMA staining, after treatment with control or the indicated doses of ketotifen (n = 4 mice / group, N = 5-6 image fields per mouse). Figure 6C shows quantification of the percentage of area positive for α-SMA staining after treatment with ketotifen compared to control. Figure 6D shows quantification of the percentage of area positive for Ki-67 staining in K7M2wt tumors after treatment with control or the indicated doses of ketotifen. Figure 6E shows representative immunofluorescence images of K7M2wt paraffin sections stained with anti-HABP1 (top row is DAPI staining, bottom row is corresponding HABP1 staining). Scale bar = 0.2 mm. Figure 6F shows quantification of the percentage of areas positive for HABP1 staining (n = 4 mice / group, N = 5-6 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≤ 0.05.

[0025] [Figure 7A-B]Figures 7A-7B show quantification of mRNA expression levels of Col1A1, CTGF, ACTA2, HAS2, HAS3, Col4, and TGFB in untreated MCA205 tumors and MCA205 tumors treated with ketotifen (10 mg / kg) (Figure 7A) and in untreated K7M2wt tumors and K7M2wt tumors treated with ketotifen (10 mg / kg) (Figure 7B) using the 2^-ΔΔCT method (3 biological replicates × 3 technical replicates were used). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the 10 mg / kg group with all other treatment groups **. p ≦ 0.05.

[0026] [Figure 8A]Figure 8A shows an exemplary study treatment protocol for the MCA205 tumor model used in Figures 8B-8H. Figures 8B-8C show relative growth curves of MCA205 tumors (n=10 mice / group) treated as indicated (Figure 8B) and K7M2wt tumors (n=6 mice / group) treated as indicated (Figure 8C). The symbol (**) indicates P<0.05 as determined by t-test by comparing the ketotifen-doxorubicin-anti-PD-L1 group with all other treatment groups. Figure 8D shows the correlation of elastic Young's modulus after 4 days of daily ketotifen in MCA205 tumors (n=5 mice / group) treated with a-PD-L1 or doxorubicin alone, or with ketotifen and ketotifen-doxorubicin-anti-PD-L1 combination therapy, to the relative tumor growth recorded by the completion of the study, day 16 (R2=0.734, p<0.0001). Figure 8E shows the correlation of elastic Young's modulus after 4 days of daily ketotifen in K7M2wt tumors (n=5 mice / group) treated with anti-PD-L1 or doxorubicin alone, or with ketotifen-doxorubicin-anti-PD-L1 combination therapy, to the relative tumor growth recorded by the completion of the study, day 33 (R2=0.857, p<0.0001). Figure 8F shows the effect on tumor burden of MCA205 tumors (n=5 mice / group) at day 16 after treatment with ketotifen alone, anti-PD-L1 alone, doxorubicin alone, ketotifen and doxorubicin combination therapy, doxorubicin and anti-PD-L1 combination, or ketotifen-doxorubicin-anti-PD-L1 combination therapy. Figure 8G shows individual tumor growth curves of surviving MCA205-bearing mice re-challenged with MCA205 tumor cells versus control mice naive to MCA205 cancer cells inoculated on day 0 with the indicated treatments (n=5 mice / group). Figure 8H shows functional perfusion areas in MCA205 tumors (n=5 mice / group) 4 days after treatment with the indicated treatments. [Fig. 8B-E]Figure 8A shows an exemplary study treatment protocol for the MCA205 tumor model used in Figures 8B-8H. Figures 8B-8C show relative growth curves of MCA205 tumors (n=10 mice / group) treated as indicated (Figure 8B) and K7M2wt tumors (n=6 mice / group) treated as indicated (Figure 8C). The symbol (**) indicates P<0.05 as determined by t-test by comparing the ketotifen-doxorubicin-anti-PD-L1 group with all other treatment groups. Figure 8D shows the correlation of elastic Young's modulus after 4 days of daily ketotifen in MCA205 tumors (n=5 mice / group) treated with a-PD-L1 or doxorubicin alone, or with ketotifen and ketotifen-doxorubicin-anti-PD-L1 combination therapy, to the relative tumor growth recorded by the completion of the study, day 16 (R2=0.734, p<0.0001). Figure 8E shows the correlation of elastic Young's modulus after 4 days of daily ketotifen in K7M2wt tumors (n=5 mice / group) treated with anti-PD-L1 or doxorubicin alone, or with ketotifen-doxorubicin-anti-PD-L1 combination therapy, to the relative tumor growth recorded by the completion of the study, day 33 (R2=0.857, p<0.0001). Figure 8F shows the effect on tumor burden of MCA205 tumors (n=5 mice / group) at day 16 after treatment with ketotifen alone, anti-PD-L1 alone, doxorubicin alone, ketotifen and doxorubicin combination therapy, doxorubicin and anti-PD-L1 combination, or ketotifen-doxorubicin-anti-PD-L1 combination therapy. Figure 8G shows individual tumor growth curves of surviving MCA205-bearing mice re-challenged with MCA205 tumor cells versus control mice naive to MCA205 cancer cells inoculated on day 0 with the indicated treatments (n=5 mice / group). Figure 8H shows functional perfusion areas in MCA205 tumors (n=5 mice / group) 4 days after treatment with the indicated treatments. [Fig. 8F-G]Figure 8A shows an exemplary study treatment protocol for the MCA205 tumor model used in Figures 8B-8H. Figures 8B-8C show relative growth curves of MCA205 tumors (n=10 mice / group) treated as indicated (Figure 8B) and K7M2wt tumors (n=6 mice / group) treated as indicated (Figure 8C). The symbol (**) indicates P<0.05 as determined by t-test by comparing the ketotifen-doxorubicin-anti-PD-L1 group with all other treatment groups. Figure 8D shows the correlation of elastic Young's modulus after 4 days of daily ketotifen in MCA205 tumors (n=5 mice / group) treated with a-PD-L1 or doxorubicin alone, or with ketotifen and ketotifen-doxorubicin-anti-PD-L1 combination therapy, to the relative tumor growth recorded by the completion of the study, day 16 (R2=0.734, p<0.0001). Figure 8E shows the correlation of elastic Young's modulus after 4 days of daily ketotifen in K7M2wt tumors (n=5 mice / group) treated with anti-PD-L1 or doxorubicin alone, or with ketotifen-doxorubicin-anti-PD-L1 combination therapy, to the relative tumor growth recorded by the completion of the study, day 33 (R2=0.857, p<0.0001). Figure 8F shows the effect on tumor burden of MCA205 tumors (n=5 mice / group) at day 16 after treatment with ketotifen alone, anti-PD-L1 alone, doxorubicin alone, ketotifen and doxorubicin combination therapy, doxorubicin and anti-PD-L1 combination, or ketotifen-doxorubicin-anti-PD-L1 combination therapy. Figure 8G shows individual tumor growth curves of surviving MCA205-bearing mice re-challenged with MCA205 tumor cells versus control mice naive to MCA205 cancer cells inoculated on day 0 with the indicated treatments (n=5 mice / group). Figure 8H shows functional perfusion areas in MCA205 tumors (n=5 mice / group) 4 days after treatment with the indicated treatments. [Figure 8H]Figure 8A shows an exemplary study treatment protocol for the MCA205 tumor model used in Figures 8B-8H. Figures 8B-8C show relative growth curves of MCA205 tumors (n=10 mice / group) treated as indicated (Figure 8B) and K7M2wt tumors (n=6 mice / group) treated as indicated (Figure 8C). The symbol (**) indicates P<0.05 as determined by t-test by comparing the ketotifen-doxorubicin-anti-PD-L1 group with all other treatment groups. Figure 8D shows the correlation of elastic Young's modulus after 4 days of daily ketotifen in MCA205 tumors (n=5 mice / group) treated with a-PD-L1 or doxorubicin alone, or with ketotifen and ketotifen-doxorubicin-anti-PD-L1 combination therapy, to the relative tumor growth recorded by the completion of the study, day 16 (R2=0.734, p<0.0001). Figure 8E shows the correlation of elastic Young's modulus after 4 days of daily ketotifen in K7M2wt tumors (n=5 mice / group) treated with anti-PD-L1 or doxorubicin alone, or with ketotifen-doxorubicin-anti-PD-L1 combination therapy, to the relative tumor growth recorded by the completion of the study, day 33 (R2=0.857, p<0.0001). Figure 8F shows the effect on tumor burden of MCA205 tumors (n=5 mice / group) at day 16 after treatment with ketotifen alone, anti-PD-L1 alone, doxorubicin alone, ketotifen and doxorubicin combination therapy, doxorubicin and anti-PD-L1 combination, or ketotifen-doxorubicin-anti-PD-L1 combination therapy. Figure 8G shows individual tumor growth curves of surviving MCA205-bearing mice re-challenged with MCA205 tumor cells versus control mice naive to MCA205 cancer cells inoculated on day 0 with the indicated treatments (n=5 mice / group). Figure 8H shows functional perfusion areas in MCA205 tumors (n=5 mice / group) 4 days after treatment with the indicated treatments.

[0027] [Figure 9A-B]Figure 9A shows the percentage of total CD3+ T cells among CD45+ lymphocytes in whole tumor tissue of MCA205 tumor models treated as indicated. Figure 9B shows the ratio of cytotoxic CD3+CD8+ T cells to CD3+CD4+CD25hiCD127loFoxp3+Treg in MCA205 tumors after the indicated treatments (n=5 mice / group). Figure 9C shows representative images of immunofluorescence (IF) staining for CD8 (bottom two rows) and Ki67 (top two rows, corresponding to the same cells as bottom two rows) in K7M2wt paraffin-embedded tissue sections. Scale bar=0.1 mm. Figure 9D shows the corresponding quantification of the percentage of proliferative CD8+ T cells as a ratio of areas double positive for CD8 and Ki67 staining to areas positive for total CD8 staining after treatment with the indicated therapies (n=4 mice / group, N=4-5 image fields per mouse). Figure 9E shows representative images of IF staining of CD31 (endothelial marker, top two rows) and CD3 (bottom two rows, corresponding to the same image fields as the top row) in K7M2wt frozen tissue sections. Scale bar = 0.1 mm. Figure 9F shows the corresponding quantification of colocalization of CD3+ T cells with CD31+ endothelial cells after the indicated treatments (n = 4 mice / group, N = 4-5 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the ketotifen-doxorubicin-anti-PD-L1 group with all other treatment groups **. p ≤ 0.05. [Figure 9C]Figure 9A shows the percentage of total CD3+ T cells among CD45+ lymphocytes in whole tumor tissue of MCA205 tumor models treated as indicated. Figure 9B shows the ratio of cytotoxic CD3+CD8+ T cells to CD3+CD4+CD25hiCD127loFoxp3+Treg in MCA205 tumors after the indicated treatments (n=5 mice / group). Figure 9C shows representative images of immunofluorescence (IF) staining for CD8 (bottom two rows) and Ki67 (top two rows, corresponding to the same cells as bottom two rows) in K7M2wt paraffin-embedded tissue sections. Scale bar=0.1 mm. Figure 9D shows the corresponding quantification of the percentage of proliferative CD8+ T cells as a ratio of areas double positive for CD8 and Ki67 staining to areas positive for total CD8 staining after treatment with the indicated therapies (n=4 mice / group, N=4-5 image fields per mouse). Figure 9E shows representative images of IF staining of CD31 (endothelial marker, top two rows) and CD3 (bottom two rows, corresponding to the same image fields as the top row) in K7M2wt frozen tissue sections. Scale bar = 0.1 mm. Figure 9F shows the corresponding quantification of colocalization of CD3+ T cells with CD31+ endothelial cells after the indicated treatments (n = 4 mice / group, N = 4-5 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the ketotifen-doxorubicin-anti-PD-L1 group with all other treatment groups **. p ≤ 0.05. [Figure 9D]Figure 9A shows the percentage of total CD3+ T cells among CD45+ lymphocytes in whole tumor tissue of MCA205 tumor models treated as indicated. Figure 9B shows the ratio of cytotoxic CD3+CD8+ T cells to CD3+CD4+CD25hiCD127loFoxp3+Treg in MCA205 tumors after the indicated treatments (n=5 mice / group). Figure 9C shows representative images of immunofluorescence (IF) staining for CD8 (bottom two rows) and Ki67 (top two rows, corresponding to the same cells as bottom two rows) in K7M2wt paraffin-embedded tissue sections. Scale bar=0.1 mm. Figure 9D shows the corresponding quantification of the percentage of proliferative CD8+ T cells as a ratio of areas double positive for CD8 and Ki67 staining to areas positive for total CD8 staining after treatment with the indicated therapies (n=4 mice / group, N=4-5 image fields per mouse). Figure 9E shows representative images of IF staining of CD31 (endothelial marker, top two rows) and CD3 (bottom two rows, corresponding to the same image fields as the top row) in K7M2wt frozen tissue sections. Scale bar = 0.1 mm. Figure 9F shows the corresponding quantification of colocalization of CD3+ T cells with CD31+ endothelial cells after the indicated treatments (n = 4 mice / group, N = 4-5 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the ketotifen-doxorubicin-anti-PD-L1 group with all other treatment groups **. p ≤ 0.05. [Figure 9E]Figure 9A shows the percentage of total CD3+ T cells among CD45+ lymphocytes in whole tumor tissue of MCA205 tumor models treated as indicated. Figure 9B shows the ratio of cytotoxic CD3+CD8+ T cells to CD3+CD4+CD25hiCD127loFoxp3+Treg in MCA205 tumors after the indicated treatments (n=5 mice / group). Figure 9C shows representative images of immunofluorescence (IF) staining for CD8 (bottom two rows) and Ki67 (top two rows, corresponding to the same cells as bottom two rows) in K7M2wt paraffin-embedded tissue sections. Scale bar=0.1 mm. Figure 9D shows the corresponding quantification of the percentage of proliferative CD8+ T cells as a ratio of areas double positive for CD8 and Ki67 staining to areas positive for total CD8 staining after treatment with the indicated therapies (n=4 mice / group, N=4-5 image fields per mouse). Figure 9E shows representative images of IF staining of CD31 (endothelial marker, top two rows) and CD3 (bottom two rows, corresponding to the same image fields as the top row) in K7M2wt frozen tissue sections. Scale bar = 0.1 mm. Figure 9F shows the corresponding quantification of colocalization of CD3+ T cells with CD31+ endothelial cells after the indicated treatments (n = 4 mice / group, N = 4-5 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the ketotifen-doxorubicin-anti-PD-L1 group with all other treatment groups **. p ≤ 0.05. [Figure 9F]Figure 9A shows the percentage of total CD3+ T cells among CD45+ lymphocytes in whole tumor tissue of MCA205 tumor models treated as indicated. Figure 9B shows the ratio of cytotoxic CD3+CD8+ T cells to CD3+CD4+CD25hiCD127loFoxp3+Treg in MCA205 tumors after the indicated treatments (n=5 mice / group). Figure 9C shows representative images of immunofluorescence (IF) staining for CD8 (bottom two rows) and Ki67 (top two rows, corresponding to the same cells as bottom two rows) in K7M2wt paraffin-embedded tissue sections. Scale bar=0.1 mm. Figure 9D shows the corresponding quantification of the percentage of proliferative CD8+ T cells as a ratio of areas double positive for CD8 and Ki67 staining to areas positive for total CD8 staining after treatment with the indicated therapies (n=4 mice / group, N=4-5 image fields per mouse). Figure 9E shows representative images of IF staining of CD31 (endothelial marker, top two rows) and CD3 (bottom two rows, corresponding to the same image fields as the top row) in K7M2wt frozen tissue sections. Scale bar = 0.1 mm. Figure 9F shows the corresponding quantification of colocalization of CD3+ T cells with CD31+ endothelial cells after the indicated treatments (n = 4 mice / group, N = 4-5 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with the control group * and the ketotifen-doxorubicin-anti-PD-L1 group with all other treatment groups **. p ≤ 0.05.

[0028] [Figure 10A]Figure 10A shows representative IF images of K7M2wt paraffin-embedded sections stained for pimonidazole adducts after pimonidazole hydrochloride injection. Scale bar = 0.2 mm (top two rows are DAPI staining and bottom two rows are corresponding hypoxic staining; top and bottom corresponding panels are the same image fields). Figure 10B shows the corresponding quantification of the percentage of hypoxic areas normalized to DAPI staining in K7M2wt tumors (n = 4 mice per treatment, N = 4-5 image fields per mouse). Figure 10C shows the relative mRNA levels of genes associated with immune cell adhesion to the endothelial wall, assessed by RT-qPCR and quantified using the 2^-ΔΔCT method, in untreated (control) MCA205 tumors and MCA205 tumors treated with 10 mg / kg ketotifen (3 biological replicates × 3 technical replicates were used). Figures 10D-10E show quantification of areas positive for CD3 (Figure 10D) and CD31 (Figure 10E) staining in K7M2wt tumor sections after the indicated treatments (n=4 mice / group, N=4-5 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups to the control group * and the ketotifen-doxorubicin-anti-PD-L1 group to all other treatment groups **. p≦0.05. [Figure 10B]Figure 10A shows representative IF images of K7M2wt paraffin-embedded sections stained for pimonidazole adducts after pimonidazole hydrochloride injection. Scale bar = 0.2 mm (top two rows are DAPI staining and bottom two rows are corresponding hypoxic staining; top and bottom corresponding panels are the same image fields). Figure 10B shows the corresponding quantification of the percentage of hypoxic areas normalized to DAPI staining in K7M2wt tumors (n = 4 mice per treatment, N = 4-5 image fields per mouse). Figure 10C shows the relative mRNA levels of genes associated with immune cell adhesion to the endothelial wall, assessed by RT-qPCR and quantified using the 2^-ΔΔCT method, in untreated (control) MCA205 tumors and MCA205 tumors treated with 10 mg / kg ketotifen (3 biological replicates × 3 technical replicates were used). Figures 10D-10E show quantification of areas positive for CD3 (Figure 10D) and CD31 (Figure 10E) staining in K7M2wt tumor sections after the indicated treatments (n=4 mice / group, N=4-5 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups to the control group * and the ketotifen-doxorubicin-anti-PD-L1 group to all other treatment groups **. p≦0.05. [Figure 10C-E]Figure 10A shows representative IF images of K7M2wt paraffin-embedded sections stained for pimonidazole adducts after pimonidazole hydrochloride injection. Scale bar = 0.2 mm (top two rows are DAPI staining and bottom two rows are corresponding hypoxic staining; top and bottom corresponding panels are the same image fields). Figure 10B shows the corresponding quantification of the percentage of hypoxic areas normalized to DAPI staining in K7M2wt tumors (n = 4 mice per treatment, N = 4-5 image fields per mouse). Figure 10C shows the relative mRNA levels of genes associated with immune cell adhesion to the endothelial wall, assessed by RT-qPCR and quantified using the 2^-ΔΔCT method, in untreated (control) MCA205 tumors and MCA205 tumors treated with 10 mg / kg ketotifen (3 biological replicates × 3 technical replicates were used). Figures 10D-10E show quantification of areas positive for CD3 (Figure 10D) and CD31 (Figure 10E) staining in K7M2wt tumor sections after the indicated treatments (n=4 mice / group, N=4-5 image fields per mouse). All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups to the control group * and the ketotifen-doxorubicin-anti-PD-L1 group to all other treatment groups **. p≦0.05.

[0029] [Figure 11A-B] Figures 11A-11B show flow cytometry analysis and gating design for analysis of MCA205 tumors. Figure 11A shows flow cytometry analysis of myeloid cells of MCA205 tumors after the indicated treatments. A myeloid gate was used on FSC / SSC to exclude dead cells and gated on CD45+ cells (Figure 11B). CD11b and Gr-1 (Ly6G+Ly6C+) antibodies were used to distinguish myeloid cell populations from monocytes, and MDSCs were identified by high expression of both CD11b and Gr-1 proteins. All data are expressed as mean ± standard error of the mean. Statistical analysis was performed by comparing treated groups with control groups *. p≦0.05.

[0030] [Figure 12]FIG. 12 shows the gating strategy for flow cytometry analysis of tumor infiltrating lymphocytes. Doublets / aggregate events were gated out using side scatter area vs. side scatter width. After gating on singlets, L / D staining was used to identify live cells, and then a lymphocyte gate (CD45+) was used to remove non-lymphoid cells. CD3+ T cells were stained with CD4 and CD8 antibodies. After gating on CD4+ cells, cells expressing low levels of CD127 and high levels of CD25 (CD127loCD25hi) were identified. This population was gated on to identify the percentage of cells that were Tregs expressing Foxp3+.

[0031] [Figure 13A-B] Figure 13A shows a co-culture assay in which mouse MC / 9 MCs and NIH3T3 fibroblasts were separated by a transwell chamber with micropores that only allowed chemical communication. Figure 13B shows the effects on mast cell degranulation by compound C48 / 80 (C48 / 80), ketotifen, and TGFβ. Figure 13C shows immunofluorescence staining of αSMA (bottom row) and collagen I (top row) in fibroblasts after the indicated treatments. Figure 13D shows quantification of collagen I and Figure 13E shows quantification of αSMA after the indicated treatments normalized to DAPI nuclear staining. Statistical analysis was performed by comparing the means between two independent groups using the usual one-way ANOVA test. [Figure 13C]Figure 13A shows a co-culture assay in which mouse MC / 9 MCs and NIH3T3 fibroblasts were separated by a transwell chamber with micropores that only allowed chemical communication. Figure 13B shows the effects on mast cell degranulation by compound C48 / 80 (C48 / 80), ketotifen, and TGFβ. Figure 13C shows immunofluorescence staining of αSMA (bottom row) and collagen I (top row) in fibroblasts after the indicated treatments. Figure 13D shows quantification of collagen I and Figure 13E shows quantification of αSMA after the indicated treatments normalized to DAPI nuclear staining. Statistical analysis was performed by comparing the means between two independent groups using the usual one-way ANOVA test. [Fig. 13D-E] Figure 13A shows a co-culture assay in which mouse MC / 9 MCs and NIH3T3 fibroblasts were separated by a transwell chamber with micropores that only allowed chemical communication. Figure 13B shows the effects on mast cell degranulation by compound C48 / 80 (C48 / 80), ketotifen, and TGFβ. Figure 13C shows immunofluorescence staining of αSMA (bottom row) and collagen I (top row) in fibroblasts after the indicated treatments. Figure 13D shows quantification of collagen I and Figure 13E shows quantification of αSMA after the indicated treatments normalized to DAPI nuclear staining. Statistical analysis was performed by comparing the means between two independent groups using the usual one-way ANOVA test.

[0032] [Figure 14A]Figure 14A shows quantification of the percentage of open lumen in MCA205 tumors based on CD31 image analysis after treatment with control or the indicated ketotifen doses. Figure 14B shows representative immunofluorescence images of NG2 pericyte marker (middle two rows) and CD31 endothelial cell marker (top two rows) of MCA205 tumors upon treatment with different ketotifen concentrations (colocalized NG2 / CD31 signals in the bottom two rows, i.e., overlapping staining). Figure 14C shows quantification of vascular pericyte coverage as indicated by overlapping staining of CD31 and NG2, normalized to total CD31+ staining. Figure 14D shows quantification of the percentage of area positive for CD31 staining (vessels) (n=5 mice / group, N=3-5 image fields per mouse). [Figure 14B] Figure 14A shows quantification of the percentage of open lumen in MCA205 tumors based on CD31 image analysis after treatment with control or the indicated ketotifen doses. Figure 14B shows representative immunofluorescence images of NG2 pericyte marker (middle two rows) and CD31 endothelial cell marker (top two rows) of MCA205 tumors upon treatment with different ketotifen concentrations (colocalized NG2 / CD31 signals in the bottom two rows, i.e., overlapping staining). Figure 14C shows quantification of vascular pericyte coverage as indicated by overlapping staining of CD31 and NG2, normalized to total CD31+ staining. Figure 14D shows quantification of the percentage of area positive for CD31 staining (vessels) (n=5 mice / group, N=3-5 image fields per mouse). [Fig. 14C-D]Figure 14A shows quantification of the percentage of open lumen in MCA205 tumors based on CD31 image analysis after treatment with control or the indicated ketotifen doses. Figure 14B shows representative immunofluorescence images of NG2 pericyte marker (middle two rows) and CD31 endothelial cell marker (top two rows) of MCA205 tumors upon treatment with different ketotifen concentrations (colocalized NG2 / CD31 signals in the bottom two rows, i.e., overlapping staining). Figure 14C shows quantification of vascular pericyte coverage as indicated by overlapping staining of CD31 and NG2, normalized to total CD31+ staining. Figure 14D shows quantification of the percentage of area positive for CD31 staining (vessels) (n=5 mice / group, N=3-5 image fields per mouse).

[0033] [Figure 15A-C] Figure 15A shows the concentration-dependent activation of MC / 9 cells by C48 / 80. Figure 15B shows the inhibitory effect of different concentrations of ketotifen on mast cell degranulation induced by C48 / 80. Figure 15C shows the assessment of NIH3T3 cell viability in the presence of ketotifen or control for 2 or 24 hours.

[0034] [Figure 16] Figure 16 shows soft tissue sarcomas infiltrated by mast cells. Immunofluorescence staining for CD117 (c-Kit) mast cell marker of human tissue arrays from patients with different sarcoma types (top row is DAPI staining, bottom row is corresponding CD117 staining).

[0035] [Figure 17A]FIG. 17A shows representative immunofluorescence images of mast cell tryptase from MCA205 tumors treated with control or the indicated doses of ketotifen (top two rows are DAPI nuclear staining, bottom two rows are corresponding tryptase staining). Scale bar is 0.05 mm. FIG. 17B and FIG. 17C show quantification of areas positive for tryptase normalized to DAPI staining for MCA205 (FIG. 17B) and K7M2wt (FIG. 17C) tumors treated with control or the indicated doses of ketotifen. [Fig. 17B-C] FIG. 17A shows representative immunofluorescence images of mast cell tryptase from MCA205 tumors treated with control or the indicated doses of ketotifen (top two rows are DAPI nuclear staining, bottom two rows are corresponding tryptase staining). Scale bar is 0.05 mm. FIG. 17B and FIG. 17C show quantification of areas positive for tryptase normalized to DAPI staining for MCA205 (FIG. 17B) and K7M2wt (FIG. 17C) tumors treated with control or the indicated doses of ketotifen.

[0036] [Figure 18A-B] Figure 18A shows normalized perfusion area relative to total tumor area at time of peak intensity for tumors following treatment with control or the indicated doses of ketotifen in the MCA205 tumor model. Figure 18B shows normalized perfusion area relative to total tumor area at time of peak intensity for tumors following completion of the treatment protocol shown in Figure 4A (n=4-5) in the K7M2wt tumor model.

[0037] [Figure 19A-B] Figure 19A shows the average elastic modulus of the tumor microscale measured using atomic force microscopy (AFM) in K7M2wt tumors (n=3 mice / group, N=10-15 different 20x20 µm force maps, grid of 16x16 points). Figure 19B shows representative AFM elastic modulus distributions / histograms of MCA205 (top and bottom left) and K7M2wt (top and bottom right) tumors after control or ketotifen treatment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0038] Detailed Description I. Definition So that the present invention may be more readily understood, certain technical and scientific terms are specifically defined below. Unless specifically defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs.

[0039] As used in this specification, including the appended claims, the singular forms of terms, e.g., "a," "an," and "the," include their corresponding plural forms unless the context clearly dictates otherwise.

[0040] A composition or method that "comprises" one or more recited elements may include other elements not specifically recited, for example, a composition that includes an antibody may contain the antibody alone or in combination with other components.

[0041] Aspects and embodiments of the invention described herein are understood to include "comprising," "consisting," and "consisting essentially of" aspects and embodiments.

[0042] A specification of a range of values ​​includes all integers within or defining the range.

[0043] Unless otherwise defined, all scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure pertains.For example, Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and the Oxford Dictionary Of Biochemistry And Molecular Biology, Revised, 2000, Oxford University Press provide those skilled in the art with a general dictionary of many of the terms used in this disclosure.

[0044] Units, prefixes, and symbols are shown in their International System of Units (SI) acceptable form. Numerical ranges include the numbers that define the range. The headings provided herein are not limitations of the various aspects of the present disclosure that can be obtained by reference to this specification as a whole. Thus, the terms defined immediately below are more fully defined by reference to this specification as a whole.

[0045] The term "weight-based dose" refers to the dose that is administered to a subject that is calculated based on the subject's body weight.For example, if a subject weighing 60 kg requires 2.0 mg / kg of ketotifen or checkpoint inhibitor, the appropriate amount (i.e., 120 mg) of ketotifen or checkpoint inhibitor can be calculated and used to administer to the subject.

[0046] The use of the term "fixed dose" in relation to the methods and dosages of the present disclosure refers to the dose administered to a subject regardless of the subject's body weight or body surface area (BSA).Therefore, the fixed dose is not provided as mg / kg dose, but as an absolute amount of agent (e.g., ketotifen and / or checkpoint inhibitor).For example, a subject weighing 60 kg and a subject weighing 100 kg are administered the same dose of ketotifen or checkpoint inhibitor.

[0047] "Cancer" refers to a broad and diverse group of diseases characterized by the uncontrolled growth of abnormal cells in the body. "Cancer" or "cancerous tissue" may include tumors. Uncontrolled cell division and growth can result in the formation of malignant tumors that invade adjacent tissues and can also metastasize to distant parts of the body through the lymphatic system or bloodstream. After metastasis, the distant tumor can be said to "originate" from the pre-metastatic tumor. For example, a "tumor originating from" breast cancer refers to a tumor that is the result of breast cancer that has metastasized.

[0048] "Administer" or "administration" refers to the physical introduction of a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those skilled in the art. Exemplary administration routes for ketotifen and / or checkpoint inhibitors include enteral administration routes, and intravenous, intramuscular, subcutaneous, intraperitoneal, intraspinal, or other parenteral administration routes, such as injection or infusion (e.g., intravenous infusion). The phrase "parenteral administration" as used herein generally refers to modes of administration other than enteral administration and topical administration by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intracisternal injection and infusion, and in vivo electroporation. The therapeutic agent can be administered via parenteral routes or orally. Other parenteral routes include topical, epidermal, or mucosal routes of administration, such as intranasal, intravaginal, rectal, sublingual, or topical. Administration can also be performed, for example, once, multiple times, and / or over one or more extended periods of time.

[0049] Reference to an amino acid residue in an antibody or other protein described herein that corresponds to a residue specified by a SEQ ID NO: includes post-translational modifications of such residue.

[0050] The term "antibody" refers to an immunoglobulin protein that is produced by the body in response to the presence of an antigen and binds to the antigen, as well as antigen-binding fragments and engineered variants thereof. Thus, the term "antibody" includes, for example, intact monoclonal antibodies (e.g., antibodies produced using hybridoma technology) and antigen-binding antibody fragments, such as F(ab')2, Fv fragments, diabodies, single-chain antibodies, scFv fragments, or scFv-Fc. In general, genetically engineered intact antibodies and fragments are also included, such as chimeric antibodies, humanized antibodies, single-chain Fv fragments, single-chain antibodies, diabodies, minibodies, linear antibodies, multivalent or multispecific (e.g., bispecific) hybrid antibodies, and the like. Thus, the term "antibody" is used broadly to include any protein that contains the antigen-binding site of an antibody and is capable of specifically binding to its antigen.

[0051] The term antibody or antigen-binding fragment thereof includes "conjugated" antibodies or antigen-binding fragments thereof, or "antibody drug conjugates (ADCs)," in which an antibody or antigen-binding fragment thereof is covalently or non-covalently linked to an agent, such as a cytotoxic agent or drug.

[0052] The term "chimeric antibody" refers to antibodies in which a portion of the heavy and / or light chain is derived from a particular species (e.g., human) or belongs to a particular antibody class or subclass, while the remainder of the chain(s) is identical or homologous to corresponding sequences in antibodies derived from another species (e.g., mouse) or belongs to another antibody class or subclass, as well as fragments of such antibodies so long as they exhibit the desired biological activity.

[0053] An "antigen-binding site of an antibody" is that portion of an antibody that is sufficient to bind to its antigen. The smallest such region is typically a variable domain or an engineered variant thereof. Single domain binding sites can be generated from camelid antibodies (Muyldermans and Lauwereys, Mol. Recog. 12: 131-140, 1999; Nguyen et al., EMBO J. 19:921-930, 2000), or VH domains of other species ("dAbs", Ward et al., Nature 341: 544-546, 1989; see U.S. Patent No. 6,248,516 to Winter et al.) to produce single domain antibodies. Generally, an antibody antigen-binding site includes both a heavy chain variable (VH) domain and a light chain variable (VL) domain that bind a common epitope. In the context of the present invention, an antibody may include one or more components in addition to the antigen binding site, such as a second antigen binding site of the antibody (which may bind to the same or a different epitope, or the same or a different antigen), a peptide linker, an immunoglobulin constant region, an immunoglobulin hinge, an amphipathic helix (see Pack and Pluckthun, Biochem. 31: 1579-1584, 1992), a non-peptide linker, an oligonucleotide (see Chaudri et al., FEBS Letters 450:23-26, 1999), a cytostatic or cytotoxic agent, etc., and may be a monomeric or multimeric protein.Examples of molecules that contain the antigen-binding site of an antibody are known in the art and include, for example, Fv, single chain Fv (scFv), Fab, Fab', F(ab')2, F(ab)c, diabodies, minibodies, nanobodies, Fab-scFv fusions, bispecific (scFv)4-IgG, and bispecific (scFv)2-Fab (see, e.g., Hu et al., Cancer Res. 56:3055-3061, 1996; Atwell et al., Molecular Immunology 33: 1301-1312, 1996; Carter and Merchant, Curr. Op. Biotechnol. 8:449-454, 1997; Zuo et al., Protein Engineering 13:361-367, 2000; and Lu et al., J. Immunol. Methods 267:213-226, (see 2002).

[0054] The term "immunoglobulin" refers to a protein consisting of one or more polypeptides substantially encoded by an immunoglobulin gene(s). One form of immunoglobulin constitutes the basic structural unit of natural (i.e., natural or parental) antibodies in vertebrates. This form is a tetramer, consisting of two identical pairs of immunoglobulin chains, each pair having one light chain and one heavy chain. In each pair, the light and heavy chain variable regions (VL and VH) together are primarily responsible for binding to the antigen, while the constant regions are primarily responsible for the antibody effector functions. In higher vertebrates, five classes of immunoglobulin proteins (IgG, IgA, IgM, IgD, and IgE) have been identified. IgG comprises the major class and is usually present as the second most abundant protein found in plasma. In humans, IgG consists of four subclasses designated IgG1, IgG2, IgG3, and IgG4. Each immunoglobulin heavy chain possesses a constant region consisting of constant region protein domains (CH1, hinge, CH2, and CH3; IgG3 also contains a CH4 domain) that are essentially invariant for a given subclass within a species.

[0055] DNA sequences encoding human and non-human immunoglobulin chains are known in the art. (e.g. Ellison et al, DNA 1: 11-18, 1981;Ellison et al, Nucleic Acids Res. 10:4071-4079, 1982;Kenten et al., Proc. Natl. Acad. Set USA 79:6661-6665, 1982;Seno et al., Nucl. Acids Res. 11 8:2055-2065, 1980;Rusconi and Kohler, Nature 314:330-334, 1985;Boss et al., Nucl. Acids Res. 12:3791-3806, 1984; Bothwell et al., Nature 298:380-382, 1982; van der Loo et al., Immunogenetics 42:333-341, 1995; Karlin et al., J. Mol. Evol. 22: 195-208, 1985; Kindsvogel et al., DNA 1 :335-343, 1982; Breiner et al., Gene 18: 165-174, 1982; Kondo et al., Eur. J. Immunol. 23:245-249, 1993; and GenBank Accession No. J00228). For reviews of immunoglobulin structure and function, see Putnam, The Plasma Proteins, Vol V, Academic Press, Inc., 49-140, 1987; and Padlan, Mol. Immunol. 31: 169-217, 1994. The term "immunoglobulin" is used herein in its general sense to refer to an intact antibody, its component chains, or fragments of the chains, depending on the context.

[0056] Full-length immunoglobulin "light chains" (about 25 kDa or 214 amino acids) are encoded at the amino terminus by a variable region gene (encoding about 110 amino acids) and at the carboxyl terminus by a kappa or lambda constant region gene. Full-length immunoglobulin "heavy chains" (about 50 kDa or 446 amino acids) are encoded by a variable region gene (encoding about 116 amino acids) and a gamma, mu, alpha, delta, or epsilon constant region gene (encoding about 330 amino acids), the latter defining the antibody's isotype, e.g., IgG, IgM, IgA, IgD, or IgE, respectively. In the light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, with the heavy chain also including a "D" region of about 10 or more amino acids. (See generally, Fundamental Immunology (Paul, ed., Raven Press, NY, 2nd ed. 1989), Ch. 7).

[0057] An immunoglobulin light or heavy chain variable region (also referred to herein as a "light chain variable domain" ("VL domain") or a "heavy chain variable domain" ("VH domain"), respectively) consists of a "framework" region interrupted by three "complementarity determining regions" or "CDRs". The framework regions act to align the CDRs to specifically bind to an epitope of an antigen. Thus, the term "CDR" refers to the amino acid residues of an antibody primarily responsible for antigen binding. From the amino to carboxyl terminus, both VL and VH domains contain the following framework (FR) and CDR regions: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0058] The assignment of amino acids to each variable region domain follows the definition in Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD, 1987 and 1991). Kabat also provides a widely used numbering convention (Kabat numbering) in which corresponding residues between different heavy chain variable regions or different light chain variable regions are assigned the same number. CDR1, 2, and 3 of the VL domain are also referred to herein as CDR-L1, CDR-L2, and CDR-L3, respectively. CDR1, 2, and 3 of the VH domain are also referred to herein as CDR-H1, CDR-H2, and CDR-H3, respectively. When so referred to, the assignment of CDRs can follow IMGT® (Lefranc et al., Developmental & Comparative Immunology 27:55-77; 2003) instead of Kabat.

[0059] Numbering of the heavy chain constant region is according to the EU index as described in Kabat (Kabat, Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, MD, 1987 and 1991).

[0060] Unless the context specifically states otherwise, the term "monoclonal antibody" is not limited to antibodies produced through hybridoma technology. The term "monoclonal antibody" can include antibodies derived from a single clone, including any eukaryotic, prokaryotic, or phage clone. In certain embodiments, the antibodies described herein are monoclonal antibodies.

[0061] "Human antibody" (HuMAb) refers to an antibody having variable regions in which both FR and CDR are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region is also derived from human germline immunoglobulin sequences. The human antibodies of the present disclosure may contain amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, the term "human antibody" as used herein is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences. The terms "human antibody" and "fully human antibody" are used interchangeably.

[0062] The term "humanized VH domain" or "humanized VL domain" refers to an immunoglobulin VH or VL domain that comprises some or all of the CDRs that are derived completely or substantially from a non-human donor immunoglobulin (e.g., mouse or rat) and a variable domain framework sequence that is derived completely or substantially from a human immunoglobulin sequence. The non-human immunoglobulin providing the CDRs is called the "donor" and the human immunoglobulin providing the framework is called the "acceptor". In some cases, humanized antibodies retain some non-human residues within the human variable domain framework regions to enhance appropriate binding characteristics (e.g., mutations in the framework may be necessary to retain binding affinity when an antibody is humanized).

[0063] A "humanized antibody" is an antibody that comprises one or both of a humanized VH domain and a humanized VL domain. Immunoglobulin constant region(s) need not be present, but if present, they are derived entirely or substantially from human immunoglobulin constant regions.

[0064] Humanized antibodies are genetically engineered antibodies in which CDRs from a non-human "donor" antibody are grafted onto human "acceptor" antibody sequences (see, e.g., Queen, U.S. Pat. Nos. 5,530,101 and 5,585,089; Winter, U.S. Pat. No. 5,225,539; Carter, U.S. Pat. No. 6,407,213; Adair, U.S. Pat. No. 5,859,205; and Foote, U.S. Pat. No. 6,881,557). The acceptor antibody sequences can be, for example, mature human antibody sequences, composites of such sequences, consensus sequences of human antibody sequences, or germline region sequences.

[0065] The human acceptor sequence can be selected for a high degree of sequence identity in the variable region framework with the donor sequence to match the typical morphology between the acceptor and donor CDRs, among other criteria. Thus, a humanized antibody is an antibody with CDRs that are completely or substantially derived from a donor antibody, and variable region framework sequences and constant regions, if present, that are completely or substantially derived from human antibody sequences. Similarly, a humanized heavy chain typically has all three CDRs that are completely or substantially derived from a donor antibody heavy chain, and heavy chain variable region framework sequences and heavy chain constant regions, if present, that are substantially derived from human heavy chain variable region framework and constant region sequences. Similarly, a humanized light chain typically has all three CDRs that are completely or substantially derived from a donor antibody light chain, and light chain variable region framework sequences and light chain constant regions, if present, that are substantially derived from human light chain variable region framework and constant region sequences.

[0066] The CDRs in a humanized antibody are substantially derived from the corresponding CDRs of a non-human antibody when at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% of the corresponding residues (as defined by Kabat numbering), or about 100% of the corresponding residues (as defined by Kabat numbering) are identical between the respective CDRs. A variable region framework sequence of an antibody chain or a constant region of an antibody chain is substantially derived from a human variable region framework sequence or a human constant region, respectively, if at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% of the corresponding residues (defined by Kabat numbering for the variable region and by EU numbering for the constant region), or about 100% of the corresponding residues (defined by Kabat numbering for the variable region and by EU numbering for the constant region) are identical.

[0067] Humanized antibodies often incorporate all six CDRs of a murine antibody (preferably as defined by Kabat or IMGT®), although they can also be generated from fewer than all six CDRs from a murine antibody (e.g., at least three, four, or five CDRs) (e.g., Pascalis et al., J. Immunol. 169:3076, 2002; Vajdos et al., Journal of Molecular Biology, 320: 415-428, 2002; Iwahashi et al., Mol. Immunol. 36:1079-1091, 1999; Tamura et al, Journal of Immunology, 164: 1432- 1441, 2000).

[0068] A CDR in a humanized antibody is "substantially derived" from a corresponding CDR of a non-human antibody if at least 60%, at least 85%, at least 90%, at least 95%, or 100% of the corresponding residues (as defined by Kabat (or IMGT)) are identical between the respective CDRs. In certain variations of humanized VH or VL domains in which the CDRs are substantially derived from a non-human immunoglobulin, the CDRs of the humanized VH or VL domain have no more than six amino acid substitutions (e.g., no more than five, no more than four, no more than three, no more than two, or no more than one) for all three CDRs compared to the corresponding non-human VH or VL CDRs (preferably conservative substitutions). The variable region framework sequence of an antibody VH or VL domain, or, if present, the sequence of the immunoglobulin constant region, is "substantially derived" from a human VH or VL framework sequence or human constant region, respectively, if at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% of the corresponding residues (as defined by Kabat numbering for the variable region and EU numbering for the constant region), or about 100% of the corresponding residues (as defined by Kabat numbering for the variable region and EU numbering for the constant region), are identical. Thus, all parts of a humanized antibody, except for the CDRs, are typically completely or substantially derived from corresponding parts of natural human immunoglobulin sequences.

[0069] The antibody is typically provided in an isolated form. This means that the antibody is at least about 50% pure by weight of interfering proteins and other contaminants resulting from its production or purification, but does not exclude the possibility that the antibody may be combined with an excess amount of pharma-ceutically acceptable carrier(s) or other medium intended to facilitate its use. Sometimes, the antibody is at least about 60%, about 70%, about 80%, about 90%, about 95%, or about 99% pure by weight of interfering proteins and contaminants resulting from its production or purification. The antibody, including the isolated antibody, can be conjugated with a cytotoxic agent and provided as an antibody drug conjugate.

[0070] Specific binding between an antibody and its target antigen typically occurs at least about 10 6 , about 10 7 , about 10 8 , about 10 9 , or about 10 10 M -1 Specific binding refers to the affinity of a molecule to a target. Specific binding is detectably higher order and is distinguished from non-specific binding that occurs to at least one non-specific target. Specific binding can be the result of the formation of bonds between specific functional groups or specific spatial fits (e.g., lock and key type), while non-specific binding is typically the result of van der Waals forces.

[0071] The term "epitope" refers to the site of an antigen to which an antibody binds. Epitopes can be formed from contiguous amino acids or non-contiguous amino acids adjacent to one or more proteins by tertiary folding. Epitopes formed from contiguous amino acids are typically retained on exposure to denaturing agents, e.g., solvents, whereas epitopes formed by tertiary folding are typically lost on treatment with denaturing agents, e.g., solvents. Epitopes typically include at least about 3, and more usually at least about 5, at least about 6, at least about 7, or about 8-10 amino acids in a unique spatial conformation. Methods for determining the spatial conformation of epitopes include, for example, x-ray crystallography and two-dimensional nuclear magnetic resonance. See, for example, Epitope Mapping Protocols, in Methods in Molecular Biology, Vol. 66, Glenn E. Morris, Ed. (1996).

[0072] Antibodies that recognize the same or overlapping epitopes can be identified in simple immunoassays that show the ability of one antibody to compete with the binding of another antibody to a target antigen. The epitope of an antibody can also be defined by X-ray crystallography of the antibody bound to its antigen to identify contact residues.

[0073] Alternatively, two antibodies have the same epitope if all amino acid mutations in the antigen that reduce or eliminate binding of one antibody also reduce or eliminate binding of the other (provided that such mutations do not result in a global alteration of the antigen structure.) Two antibodies have overlapping epitopes if some amino acid mutations that reduce or eliminate binding of one antibody also reduce or eliminate binding of the other antibody.

[0074] Competition between antibodies can be determined by an assay in which a test antibody inhibits specific binding of a reference antibody to a common antigen (see, e.g., Junghans et al., Cancer Res. 50: 1495, 1990). A test antibody competes with a reference antibody if an excess of the test antibody inhibits binding of the reference antibody.

[0075] Antibodies identified by competitive assays (competitor antibodies) include antibodies that bind to the same epitope as the reference antibody, and antibodies that bind to adjacent epitopes sufficiently proximal to the epitope bound by the reference antibody such that steric interference occurs. Antibodies identified by competitive assays also include antibodies that indirectly compete with the reference antibody by inducing a conformational change in the target protein, thereby preventing the reference antibody from binding to an epitope different from the epitope bound by the test antibody.

[0076] Antibody effector function refers to a function involving the Fc region of Ig. Such a function can be, for example, antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), or complement-dependent cytotoxicity (CDC). Such a function can be influenced, for example, by the binding of the Fc region to Fc receptors on immune cells with phagocytic or lytic activity, or by the binding of the Fc region to components of the complement system. Typically, the effect(s) mediated by Fc-binding cells or complement components results in the inhibition and / or depletion of LIV1-targeted cells. The Fc region of an antibody can recruit Fc receptor (FcR)-expressing cells and juxtapose them with antibody-coated target cells. Cells expressing surface FcRs of IgG, including FcγRIII (CD16), FcγRII (CD32), and FcγRIII (CD64), can act as effector cells to destroy IgG-coated cells. Such effector cells include monocytes, macrophages, natural killer (NK) cells, neutrophils, and eosinophils. Engagement of FcγR with IgG activates ADCC or ADCP. ADCC is mediated by CD16+ effector cells through secretion of pore-forming proteins and proteases, whereas phagocytosis is mediated by CD32+ and CD64+ effector cells (Fundamental Immunology, 4 th ed., Paul ed., Lippincott-Raven, NY, 1997, Chapters 3, 17 and 30;Uchida et al., J. Exp. Med. 199:1659-69, 2004;Akewanlop et al., Cancer Res. 61:4061-65, 2001;Watanabe et al., Breast Cancer Res. Treat. 53: 199-207, 1999).

[0077] In addition to ADCC and ADCP, the Fc region of cell-bound antibodies can also activate the classical complement pathway, triggering CDC. C1q of the complement system binds to the Fc region of antibodies when they form a complex with an antigen. Binding of C1q to cell-bound antibodies can initiate a cascade of events involving proteolytic activation of C4 and C2 to generate C3 convertase. Cleavage of C3 to C3b by C3 convertase allows activation of the final complement components, including C5b, C6, C7, C8, and C9. Collectively, these proteins form membrane attack complex pores on antibody-coated cells. These pores disrupt the integrity of the cell membrane, killing the target cell (Immunobiology, 6 th ed., Janeway et al, Garland Science, NY, 2005, Chapter 2).

[0078] The term "antibody-dependent cellular cytotoxicity" or "ADCC" refers to a mechanism of inducing cell death that depends on the interaction of antibody-coated target cells with immune cells (also called effector cells) that possess lytic activity. Such effector cells include natural killer cells, monocytes / macrophages, and neutrophils. Effector cells bind to the Fc region of Ig bound to target cells via their antigen-binding sites. Effector cell activity results in the killing of antibody-coated target cells. In certain exemplary embodiments, the anti-LIV1 IgG1 antibodies of the invention mediate equal or increased ADCC compared to parental antibodies and / or compared to anti-LIV1 IgG3 antibodies.

[0079] The term "antibody-dependent cellular phagocytosis" or "ADCP" refers to the process by which antibody-coated cells are internalized in whole or in part by phagocytic immune cells (e.g., macrophages, neutrophils, and / or dendritic cells) that bind the Fc region of Ig. In certain exemplary embodiments, the anti-LIV1 IgG1 antibodies of the invention mediate equal or increased ADCP compared to the parent antibody and / or compared to an anti-LIV1 IgG3 antibody.

[0080] The term "complement dependent cytotoxicity" or "CDC" refers to a mechanism in which the Fc region of a target-bound antibody activates a series of enzymatic reactions that create holes in the target cell membrane, inducing cell death.

[0081] Typically, antigen-antibody complexes, such as those on antibody-coated target cells, bind to and activate complement component C1q, which then activates the complement cascade, resulting in target cell death. Complement activation also results in the deposition of complement components on the target cell surface, which can facilitate ADCC by binding to complement receptors (e.g., CR3) on leukocytes.

[0082] "Cytotoxicity" refers to the depletion, elimination, and / or killing of target cells. "Cytotoxic agent" refers to a compound that has a cytotoxic effect on cells, thereby mediating the depletion, elimination, and / or killing of target cells. In certain embodiments, the cytotoxic agent is conjugated to or administered in combination with an antibody. Suitable cytotoxic agents are further described herein.

[0083] "Cytostatic effect" refers to the inhibition of cell proliferation. "Cytostatic agent" refers to a compound that has a cytostatic effect on cells, thereby mediating the inhibition of growth and / or expansion of a particular cell type and / or cell subset. Suitable cytostatic agents are further described herein.

[0084] As used herein, a "sub-therapeutic dose" means a dose of a therapeutic compound (e.g., ketotifen or a checkpoint inhibitor) that is lower than the usual or typical dose of the therapeutic compound when administered alone for the treatment of a hyperproliferative disease (e.g., cancer) and / or, in the case of ketotifen, lower than the usual or typical dose used to treat the indicated disease (e.g., pulmonary hypertension).

[0085] As an example, an "anti-cancer drug" promotes the regression of cancer in a subject. In some embodiments, a therapeutically effective amount of a drug promotes the regression of cancer to the point of eliminating the cancer. "Promoting the regression of cancer" means that administering an effective amount of a drug, alone or in combination with an anti-cancer drug, results in a reduction in tumor growth or size, tumor necrosis, a decrease in the severity of at least one disease symptom, an increase in the number and duration of disease-free periods, or prevention of functional impairment or disability due to disease morbidity. In addition, the terms "effective" and "efficacy" in relation to treatment include both pharmacological efficacy and physiological safety. Pharmacological efficacy refers to the ability of a drug to promote the regression of cancer in a patient. Physiological safety refers to the level of toxicity or other adverse physiological effects (adverse effects) at the cell, organ, and / or organism level resulting from the administration of the drug.

[0086] A "chemotherapeutic agent" is a chemical compound useful in the treatment of cancer. Examples of chemotherapeutic agents include alkylating agents, such as thiotepa and cyclophosphamide (CYTOXAN®); alkylsulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylameramines, such as altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; acetogens, such as cyclosporine, cyclopentasiloxane, cyclohex ... camptothecins (especially bullatacin and bullatacinone); delta-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapachone; lapachol; colchicine; betulinic acid; camptothecins (including the synthetic analogs topotecan (HYCAMTIN®), CPT-11 (irinotecan, CAMPTOSAR®), acetylcamptothecin, scopolectin, and 9-aminocamptothecin); bryostatin; kallistatin; CC-1065 (its including synthetic analogs of adozelesin, carzelesin, and bizelesin; podophyllotoxins; podophyllic acid; teniposide; cryptophycins (especially cryptophycin 1 and cryptophycin 8); dolastatins; duocarmycins (including synthetic analogs, KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictin; spongiostatin; nitrogen mustards, e.g. chlorambucil, chlornaphazine, chlorophosphamide, estramustine , ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobembicine, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics, such as enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gamma II and calicheamicin omega II (see, e.g., Nicolaou et al., Angew. Chem Intl. Ed. Engl., 33: 183-186 (1994));CDP323, an oral alpha-4 integrin inhibitor; dynemicins, e.g., dynemicin A; esperamicin; and neocarzinostatin chromophores and related chromoprotein enediyne antibiotic chromophores), aclacinomycin, actinomycin, ausramycin, azaserine, bleomycin, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (e.g., ADRIAMYCIN®, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, doxorubicin HCl liposome injection (DOXIL®), liposomal doxorubicin TLC D-99 (including MYOCET®, pegylated liposomal doxorubicin (CAELYX®, and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcelomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, queramycin, lodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate, gemcitabine (GEMZAR®), tegafur (UFTORAL®), capecitabine (XELODA®), epothilones, and 5-furan fluorouracil (5-FU); combretastatins; folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calsterone, dromostanol propionate, epithiostanol, mepitiostane, testolactone; antiadrenal agents such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as floric acid; aceglatone; aldophosphamide glycosides;Aminolevulinic acid; Eniluracil; Amsacrine; Bestravcil; Bisantrene; Edatrexate; Defofamine; Demecolcine; Diazicon; Elformithine; Elliptinium acetate; Epothilone; Etoglucide; Gallium nitrate; Hydroxyurea; Lentinan; Lonidynin; Maytansinoids, such as maytansine and ansamitocin; Mitoguazone; Mitoxantrone; Mopidanmol; Nitraerin; Pentostatin; Phenamet; Pirarubicin; Rosoxantrone; 2-Ethylhydrazide; Procarbazine; PSK® Polysaccharide Complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; schizofuran; spirogermanium; tenuazonic acid; triazicon; 2,2',2'-trichlorotriethylamine; trichothecenes (especially T-2 toxin, veracrine A, roridin A, and anguidine); urethanes; vindesine (ELDISINE®, FILDESIN®); dacarbazine; mannommustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); thiotepa; taxoids such as paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, NJ), albumin engineered nanoparticle formulations of paclitaxel (ABRAXANE™), and docetaxel (TAXOTERE®, Rhome-Poulene Rorer, Antony, France); chlorambucil; 6-thioguanine; mercaptopurine; methotrexate; platinum agents such as cisplatin, oxaliplatin (e.g., ELOXATIN®), and carboplatin; vincas, which prevent tubulin polymerization to form microtubules, such as vinblastine (VELBAN®), vincristine (ONCOVIN®), vindesine (ELDISINE®, FILDESIN®), and vinorelbine (NAVELBINE®); etoposide (VP-16); ifosfamide; mitoxantrone; leucovorin; novantrone; edatrexate; daunomycin;Aminopterin; ibandronate; topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoids, such as retinoic acid, such as bexarotene (TARGRETIN®); bisphosphonates, such as clodronate (e.g., BONEFOS® or OSTAC®), etidronate (DIDROCAL®), NE-58095, zoledronic acid / zoledronate (ZOMETA®), alendronate (FOSAMAX®), paclitaxel, midronate (AREDIA®), tiludronate (SKELID®), or risedronate (ACTONEL®); troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); antisense oligonucleotides, particularly those that inhibit the expression of genes in signal transduction pathways involved in abnormal cell proliferation, such as PKC-alpha, Raf, H-Ras, and epidermal growth factor receptor (EGF-R) (e.g., erlotinib (Tarceva™)); and VEGF-A etc., which reduce cell proliferation; vaccines, such as THERATOOPE® vaccine and gene therapy vaccines, such as ALLOVECTIN® vaccine, LEUVECTIN® vaccine, and VAXID® vaccine; topoisomerase 1 inhibitors (e.g., LURTOTECAN®); rmRH (e.g., ABARELIX®); BAY439006 (sorafenib; Bayer); SU-11248 sunitinib, SUTENT®, Pfizer) ; Perifosine, COX-2 inhibitors (e.g., celecoxib or etoricoxib), proteosome inhibitors (e.g., PS341); bortezomib (VELCADE®); CCI-779; tipifarnib (R11577); orafenib, ABT510; Bcl-2 inhibitors, e.g., oblimersen sodium (GENASENSE®); pixantrone; EGFR inhibitors; tyrosine kinase inhibitors; serine-threonine kinase inhibitors, e.g., rapamycin (sirolimus, RAPAMUNE®);Farnesyltransferase inhibitors, such as lonafarnib (SCH 6636, SARASAR™); checkpoint inhibitors (e.g., CTLA-4, PD-1, PD-L1, PD-L2, B7-H3, B7-H4, BMA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, or inhibitors of the B-7 family ligand); and pharmaceutically acceptable salts, acids, or derivatives of any of the above; and combinations of two or more of the above, such as CHOP, which is an abbreviation for combination therapy of cyclophosphamide, doxorubicin, vincristine, and prednisolone; and FOLFOX, which is an abbreviation for a treatment regimen with oxaliplatin in combination with 5-FU and leucovorin (ELOXATIN™), and pharmaceutically acceptable salts, acids, or derivatives of any of the above; and combinations of two or more of the above.

[0087] Chemotherapeutic agents, as defined herein, include "anti-hormonal agents" or "endocrine therapeutic agents" that act to regulate, reduce, block, or inhibit the effects of hormones that may promote cancer growth. These include hormonal agents themselves, including, but not limited to, antiestrogens and selective estrogen receptor modulators (SERMs), such as tamoxifen (including NOLVADEX® tamoxifen), raloxifene, droloxifene, 4-hydroxytamoxifen, trioxyphene, keoxyphene, LY117018, onapristone, and FARESTON® toremifene; aromatase inhibitors that inhibit the enzyme aromatase, which regulates estrogen production in the adrenal glands, such as 4(5)-imidazole, aminoglutethimide, MEGASE® megestrol acetate, AROMASIN® exemestane, formestane, fadrozole, RIVISOR® vorozole, FEMARA® letrozole, and ARIMIDEX® anastrozole; and antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and gonoside. sereline; and troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); antisense oligonucleotides, particularly those that inhibit the expression of genes in signal transduction pathways involved in abnormal cell proliferation, such as PKC-alpha, Raf, and H-Ras; ribozymes, such as VEGF expression inhibitors (e.g., ANGIOZYME® ribozymes) and HER2 expression inhibitors; vaccines, such as gene therapy vaccines, such as the ALLOVECTIN® vaccine, the LEUVECTIN® vaccine, and the VAXID® vaccine; PROLEUKIN® rlL-2; LURTOTECAN® topoisomerase 1 inhibitors; ABARELIX® rmRH; vinorelbine and esperamicin (see U.S. Pat. No. 4,675,187), as well as pharmaceutically acceptable salts, acids, or derivatives of any of the above; and combinations of two or more of the above.

[0088] The term "baseline" or "baseline value" is used interchangeably herein and may refer to a measurement or characterization of a symptom before administration of a treatment (e.g., ketotifen or its pharma- ceutically acceptable salts described herein and / or a checkpoint inhibitor described herein) or at the start of administration of a treatment. The baseline value may be compared to a reference value to determine a reduction or improvement in symptoms of a disease, such as cancer. The term "reference" or "reference value" is used interchangeably herein and may refer to a measurement or characterization of a symptom after administration of a treatment (e.g., ketotifen or its pharma-ceutically acceptable salts described herein and / or a checkpoint inhibitor described herein). The reference value may be measured one or more times during a dosing regimen or treatment cycle, or at the completion of a dosing regimen or treatment cycle. The "reference value" may be an absolute value, a relative value, a value with upper and / or lower limits, a range of values, an average value, a median value, a mean value, or a value compared to a baseline value.

[0089] Similarly, a "baseline value" can be an absolute value, a relative value, a value with upper and / or lower limits, a range of values, an average value, a median value, a mean value, or a value compared to a reference value. Reference and / or baseline values ​​can be obtained from one individual, from two different individuals, or from a group of individuals (e.g., a group of 2, 3, 4, 5, or more individuals).

[0090] "Sustained response" refers to a sustained effect on reducing tumor growth after the cessation of treatment. For example, the tumor size may remain the same or smaller compared to the size at the beginning of the administration phase. In some embodiments, the sustained response has a duration that is at least the same as the treatment period, or has a duration that is at least 1.5, 2.0, 2.5, or 3 times longer than the treatment period.

[0091] As used herein, "complete response" or "CR" refers to the disappearance of all target lesions, "partial response" or "PR" refers to at least a 30% decrease in the sum of the longest diameters (SLD) of the target lesions, referenced to the baseline SLD; "stable disease" or "SD" refers to neither a sufficient shrinkage of the target lesions to meet the criteria for PR nor a sufficient increase, referenced to the smallest SLD after the start of treatment, to meet the criteria for PD.

[0092] As used herein, "progression-free survival" or "PFS" refers to the period during and after treatment during which the disease being treated (e.g., cancer) does not worsen. Progression-free survival can include the length of time during which a patient experiences a complete or partial response, as well as the length of time during which a patient experiences stable disease.

[0093] As used herein, "objective response rate" or "ORR" refers to the combined complete response (CR) rate and partial response (PR) rate.

[0094] As used herein, "overall survival" or "OS" refers to the percentage of individuals in a group who are likely to be alive after a particular period of time.

[0095] The term "patient" or "subject" includes human and other mammalian subjects, such as non-human primates, rabbits, rats, mice, etc., and transgenic species thereof, that receive either prophylactic or therapeutic treatment.

[0096] The term "effective amount" in the context of treating solid tumors by administering ketotifen and / or checkpoint inhibitors as described herein refers to an amount of such ketotifen and / or checkpoint inhibitor that is sufficient to inhibit or ameliorate the development of one or more symptoms of solid tumors. An effective amount of the antibody is administered in an "effective regimen". The term "effective regimen" refers to the combination of the amount of ketotifen and / or checkpoint inhibitor administered and the dosing frequency that is appropriate to achieve preventive or therapeutic treatment of a disorder (e.g., preventive or therapeutic treatment of a solid tumor).

[0097] The term "pharmaceutical acceptable" means approved or approvable by a regulatory agency of the Federal or State government, or listed in the United States Pharmacy or other generally recognized pharmacopoeias, for use in animals, and more particularly in humans. The term "pharmaceutical compatible ingredient" refers to a pharmaceutical acceptable diluent, adjuvant, excipient, or vehicle in which ketotifen or a checkpoint inhibitor is formulated.

[0098] The phrase "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable organic or inorganic salt. Exemplary salts include sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, hydrogen sulfate, phosphate, superphosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, acid tartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1' -methylenebis-(2-hydroxy-3-naphthoate). Pharmaceutically acceptable salts can also further include additional molecules, such as acetate, succinate, or other counterions. Counterions can be any organic or inorganic moiety that stabilizes the charge on the parent compound. In addition, pharmaceutically acceptable salts can have more than one charged atom in their structure. Examples where multiple charged atoms are part of a pharmaceutically acceptable salt can have multiple counterions. Thus, pharmaceutically acceptable salts can have one or more charged atoms and / or one or more counterions.

[0099] The use of the alternative (e.g., "or") should be understood to mean either one, both, or any combination thereof of the alternatives. As used herein, the indefinite article "a" or "an" should be understood to refer to "one or more" of any recited or listed components.

[0100] The term "and / or" as used herein should be understood as a specific disclosure of each of the two specified features or components with or without the other. Thus, when the term "and / or" is used herein in phrases such as "A and / or B," it is intended to include "A and B," "A or B," "A" (single), and "B" (single). Similarly, when the term "and / or" is used in phrases such as "A, B, and / or C," it is intended to encompass each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (single); B (single); and C (single).

[0101] The term "about" or "essentially comprising" refers to a value or composition that is within an acceptable error range for a particular value or composition as determined by one of ordinary skill in the art, which depends in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, "about" or "essentially comprising" can mean within one standard deviation or more than one standard deviation, according to the practice in the art. Alternatively, "about" or "essentially comprising" can mean a range of up to 20%. Furthermore, particularly with respect to biological systems or processes, the term can mean up to an order of magnitude, or up to five times the value. When a particular value or composition is provided in the present application and claims, unless otherwise stated, the meaning of "about" or "essentially comprising" should be assumed to be within an acceptable error range for that particular value or composition.

[0102] Solvate in the context of the present invention is the form of the compound of the present invention that forms a complex in solid or liquid state through coordination with solvent molecules.Hydrate is one particular form of solvate in which coordination occurs with water.In certain exemplary embodiments, solvate in the context of the present invention is hydrate.

[0103] The term "inhibit" or "inhibition of" means to reduce by a measurable amount or to prevent completely. The term inhibit, as used herein, can refer to an inhibition or reduction of at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%.

[0104] The term "treatment" or "treating" refers to slowing, halting, or reversing the progression of a disease or condition in a patient, as evidenced by a reduction or elimination of clinical or diagnostic symptoms of the disease or condition. Treatment can include, for example, a reduction in the severity of symptoms, the number of symptoms, or the frequency of recurrence.

[0105] The term "prodrug" as used herein refers to a compound that is converted to the active form of the compound upon administration in vivo. For example, the prodrug form of the active compound may be, but is not limited to, the acylation (acetylation or otherwise) and ether derivatives, carboxylate or phosphate esters, and various salt forms of the active compound. Those skilled in the art will recognize how to easily modify the compounds of the present invention into prodrug forms to facilitate the delivery of the active compound to a target site within a host organism or patient. Those skilled in the art will also take advantage of the favorable pharmacokinetic parameters of the prodrug form, if applicable, to deliver the desired compound to a target site within a host organism or patient to maximize the intended effect of the compound in the treatment of cancer.

[0106] As used herein, the terms "synergy" or "synergistic effect," when used in connection with describing the effectiveness of a combination of agents, means any measured effect of the combination that is greater than that expected from the sum of the effects of the individual agents.

[0107] As used herein, the terms "additive" or "additive effect" when used in connection with describing the effectiveness of a combination of agents means any measured effect of the combination that is similar to that expected from the sum of the effects of the individual agents.

[0108] The terms "about once every week," "about once every two weeks," or any other similar dosing interval term used herein, are meant to be approximate. "About once every week" can include every 7 days ± 1 day, i.e., every 6 to 8 days. "About once every two weeks" can include every 14 days ± 2 days, i.e., every 12 to 16 days. "About once every three weeks" can include every 21 days ± 3 days, i.e., every 18 to 24 days. Similar approximations apply, for example, to about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, and about once every 12 weeks. In some embodiments, a dosing interval of about once every 6 weeks or about once every 12 weeks means that the first dose can be administered on any day in the first week, and then the next dose can be administered on any day in the sixth or twelfth week, respectively. In other embodiments, a dosing interval of about once every 6 weeks or about once every 12 weeks means that the first dose is administered on a particular day (e.g., Monday) in the first week, and then the next dose is administered on the same day (i.e., Monday) in the 6th or 12th week, respectively.

[0109] As described herein, any concentration range, percentage range, ratio range, or integer range should be understood to include any integer value within the recited range, and, unless otherwise indicated, to include decimals thereof (e.g., tenths and hundredths of integers), where applicable.

[0110] Various aspects of the disclosure are described in further detail in the following subsections.

[0111] II. Ketotifen The compound ketotifen is a relatively selective, non-competitive histamine antagonist (H1-receptor) and mast cell stabilizer. Ketotifen is known to inhibit the release of mediators in hypersensitivity reactions. Ketotifen has been administered in both oral and ophthalmic forms. Ketotifen has the following formula: [ka]

[0112] In some embodiments, ketotifen as used herein has the formula: [ka] This refers to salts such as salts with fumaric acid and ketotifen fumarate, which have the formula:

[0113] In some embodiments of any of the methods described, ketotifen is provided as a pharma- ceutically acceptable salt of ketotifen. Preparations of ketotifen are disclosed, for example, in WO2010 / 107525A1, WO2009 / 136903A1, US2010 / 160293A1, and WO2006 / 047418A1.

[0114] III. Checkpoint Inhibitors Immune checkpoints refer to inhibitory pathways of the immune system involved in maintaining self-tolerance and modulating the degree of immune system response to minimize peripheral tissue damage. However, tumor cells can also activate immune system checkpoints to reduce the effectiveness of the immune response against tumor tissue ("blocking" the immune response). In contrast to many anticancer drugs, checkpoint inhibitors do not directly target tumor cells, but rather target lymphocyte receptors or their ligands to enhance the intrinsic antitumor activity of the immune system (Pardoll, 2012, Nature Reviews Cancer 12:252-264). Treatment with antagonistic checkpoint blocking antibodies against immune system checkpoints, such as CTLA4, PD1, and PD-L1, is one of the most promising new avenues of immunotherapy for cancer and other diseases. Additional checkpoint targets, such as TIM-3, LAG-3, various B-7 ligands, CHK1 and CHK2 kinases, BTLA, A2aR, and others, are also under testing. Checkpoint inhibitors include atezolizumab (Tecentriq®), a PD-L1 inhibitor, ipilimumab (Yervoy®), a CTLA-4 inhibitor, and pembrolizumab (Keytruda®) and nivolumab (Opdivo®), both of which are PD-1 inhibitors.

[0115] Recent data suggests a possible secondary mechanism of anti-CTLA-4 antibodies within the tumor itself. CTLA-4 has been found to be expressed at higher levels in regulatory T cells (also referred to herein as "Treg cells") compared to intratumoral effector T cells (also referred to herein as "Teff cells") in tumors, leading to the hypothesis that anti-CTLA-4 preferentially affects Treg cells.

[0116] One mechanism by which checkpoint blocking anti-CTLA-4 antibodies mediate their antitumor effects is by reducing regulatory T cells. The distinct mechanism of action of anti-CTLA-4 antibodies allows them to be successfully combined with anti-PD-1 checkpoint blocking antibodies, which act to release inhibitory signals conferred to effector T cells. Combining dual blockade with these antibodies improves antitumor responses both preclinically (Proc Natl Acad Sci USA 2010, 107, 4275-4280) and clinically (N Engl J Med 2013, 369, 122-133;N Engl J Med 2015, 372, 2006-2017).

[0117] In some embodiments, the checkpoint inhibitor inhibits a checkpoint protein selected from the group consisting of CTLA-4, PD-1, PD-L1, PD-L2, B7-H3, B7-H4, BMA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, and B-7 family ligand, or a combination thereof. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein CTLA-4. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein PD-1. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein PD-L1. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein PD-L2. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein B7-H3. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein B7-H4. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein BMA. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein HVEM. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein TIM3. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein GAL9. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein LAG3. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein VISTA. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein KIR. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein 2B4. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein CD160. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein CGEN-15049. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein CHK1.In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein CHK2. In some embodiments, the checkpoint inhibitor inhibits the checkpoint protein A2aR. In some embodiments, the checkpoint inhibitor inhibits a B-7 family ligand. In some embodiments, the checkpoint is an antibody. In some embodiments, the checkpoint inhibitor is an anti-CTLA4 antibody. In some embodiments, the checkpoint inhibitor is an anti-PD-1 antibody. In some embodiments, the checkpoint inhibitor is an anti-PD-L1 antibody. In some embodiments, the checkpoint inhibitor is an anti-PD-L2 antibody. In some embodiments, the checkpoint inhibitor is an anti-B7-H3 antibody. In some embodiments, the checkpoint inhibitor is an anti-B7-H4 antibody. In some embodiments, the checkpoint inhibitor is an anti-BMA antibody. In some embodiments, the checkpoint inhibitor is an anti-HVEM antibody. In some embodiments, the checkpoint inhibitor is an anti-TIM3 antibody. In some embodiments, the checkpoint inhibitor is an anti-GAL9 antibody. In some embodiments, the checkpoint inhibitor is an anti-LAG3 antibody. In some embodiments, the checkpoint inhibitor is an anti-VISTA antibody. In some embodiments, the checkpoint inhibitor is an anti-KIR antibody. In some embodiments, the checkpoint inhibitor is an anti-2B4 antibody. In some embodiments, the checkpoint inhibitor is an anti-CD160 antibody. In some embodiments, the checkpoint inhibitor is an anti-CGEN-15049 antibody. In some embodiments, the checkpoint inhibitor is an anti-CHK1 antibody. In some embodiments, the checkpoint inhibitor is an anti-CHK2 antibody. In some embodiments, the checkpoint inhibitor is an anti-A2aR antibody. In some embodiments, the checkpoint inhibitor is an anti-B7 family ligand antibody. In some embodiments, the checkpoint inhibitor described herein is a monoclonal antibody. In some embodiments, the checkpoint inhibitor described herein is a human antibody. In some embodiments, the checkpoint inhibitor described herein is a humanized antibody.In some embodiments, the checkpoint inhibitors described herein are chimeric antibodies. In some embodiments, the checkpoint inhibitors described herein are full-length antibodies. In some embodiments, the checkpoint inhibitors described herein are antigen-binding fragments of antibodies. In some embodiments, the antigen-binding fragments are Fab, Fab', and F(ab')2, Fd, single-chain Fv (scFv), single-chain antibodies, disulfide-linked Fv (sdFv), and V. L or V H In some embodiments, the checkpoint inhibitors described herein are selected from the group consisting of MEDI0680, AMP-224, nivolumab, pembrolizumab, pidilizumab, MEDI4736, atezolizumab, ipilimumab, tremelimumab, and BMS-936559. In some embodiments, the CDRs are Kabat CDRs. Kabat et al. (1991), "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat" numbering scheme). In some embodiments, the checkpoint inhibitors described herein are selected from the group consisting of MEDI0680, AMP-224, nivolumab, pembrolizumab, pidilizumab, MEDI4736, atezolizumab, ipilimumab, tremelimumab, and BMS-936559. The checkpoint inhibitors described herein comprise a heavy chain variable region and / or a light chain variable region of an antibody selected from the group consisting of atezolizumab, ipilimumab, tremelimumab, and BMS-936559. In some embodiments, the checkpoint inhibitors described herein comprise a heavy chain variable region of an antibody selected from the group consisting of MEDI0680, AMP-224, nivolumab, pembrolizumab, pidilizumab, MEDI4736, atezolizumab, ipilimumab, tremelimumab, and BMS-936559. In some embodiments, the checkpoint inhibitors described herein comprise a light chain variable region of an antibody selected from the group consisting of MEDI0680, AMP-224, nivolumab, pembrolizumab, pidilizumab, MEDI4736, atezolizumab, ipilimumab, tremelimumab, and BMS-936559. In some embodiments, the checkpoint inhibitors described herein comprise a heavy chain variable region and a light chain variable region of an antibody selected from the group consisting of MEDI0680, AMP-224, nivolumab, pembrolizumab, pidilizumab, MEDI4736, atezolizumab, ipilimumab, tremelimumab, and BMS-936559. In some embodiments, the checkpoint inhibitors described herein are antibodies selected from the group consisting of MEDI0680, AMP-224, nivolumab, pembrolizumab, pidilizumab, MEDI4736, atezolizumab, ipilimumab, tremelimumab, and BMS-936559. In some embodiments, the checkpoint inhibitors described herein are bioanalogs of antibodies selected from the group consisting of MEDI0680, AMP-224, nivolumab, pembrolizumab, pidilizumab, MEDI4736, atezolizumab, ipilimumab, tremelimumab, and BMS-936559. In some embodiments, the checkpoint inhibitors described herein are MEDI0680. In some embodiments, the checkpoint inhibitors described herein are AMP-224. In some embodiments, the checkpoint inhibitors described herein are nivolumab. In some embodiments, the checkpoint inhibitors described herein are pembrolizumab.In some embodiments, the checkpoint inhibitor described herein is pidilizumab. In some embodiments, the checkpoint inhibitor described herein is MEDI4736. In some embodiments, the checkpoint inhibitor described herein is atezolizumab. In some embodiments, the checkpoint inhibitor described herein is ipilimumab. In some embodiments, the checkpoint inhibitor described herein is tremelimumab. In some embodiments, the checkpoint inhibitor described herein is BMS-936559. In some embodiments, the checkpoint inhibitor is a combination of an anti-PD-1 antibody and an anti-CTLA4 antibody. In some embodiments, the checkpoint inhibitor is a combination of nivolumab and ipilimumab. In some embodiments, the checkpoint inhibitor is a combination of pembrolizumab and ipilimumab. In some embodiments, the checkpoint inhibitor is a combination of an anti-PD-L1 antibody and an anti-CTLA4 antibody. In some embodiments, the checkpoint inhibitor is a combination of atezolizumab and ipilimumab.

[0118] IV. Method A. Treatment of Solid Tumors One aspect of the present invention provides a method of treating a solid tumor in a subject in need thereof, comprising administering to the subject an effective amount of ketotifen or a pharma- ceutically acceptable salt thereof in combination with a checkpoint inhibitor. In another aspect, the present invention provides a method of initiating, enhancing, or prolonging the effect of, or enabling a subject to respond to, a checkpoint inhibitor in a subject in need thereof, comprising administering to the subject an effective amount of ketotifen or a pharma- ceutically acceptable salt thereof in combination with a checkpoint inhibitor, wherein the subject has a solid tumor. In another aspect, the present invention provides a method of enhancing the effect of a checkpoint inhibitor in a subject in need thereof, comprising administering to the subject an effective amount of ketotifen or a pharma- ceutically acceptable salt thereof in combination with a checkpoint inhibitor, wherein the subject has a solid tumor. Also provided herein is a method of increasing blood flow of a solid tumor in a subject, comprising administering to the subject an effective amount of ketotifen or a pharma- ceutically acceptable salt thereof in combination with a checkpoint inhibitor, wherein increasing blood flow of the solid tumor enhances the effect of the checkpoint inhibitor. In some embodiments, the blood flow of the solid tumor is determined using ultrasound-based blood flow measurements or using histology techniques to measure hypoxia. In some embodiments, the blood flow of the solid tumor is determined using ultrasound-based blood flow measurements. In some embodiments, the blood flow of the solid tumor is determined using histology techniques to measure hypoxia. In some embodiments, the blood flow is measured using histology techniques to measure hypoxia in a biopsy from a solid tumor. In another aspect, the invention provides a method of improving the delivery or effectiveness of a checkpoint inhibitor in a subject, comprising administering an effective amount of ketotifen or a pharma- ceutically acceptable salt thereof in combination with a checkpoint inhibitor, the subject having a solid tumor, thereby improving the delivery or effectiveness of the treatment in the subject. In some embodiments, the subject is a human.In some embodiments, the method further comprises administering an effective amount of ketotifen, or a pharma- ceutically acceptable salt thereof, in combination with an effective amount of an additional chemotherapeutic agent, such as doxorubicin. In some embodiments, the method further comprises administering an effective amount of ketotifen, or a pharma- ceutically acceptable salt thereof, in combination with an effective amount of an additional chemotherapeutic agent, such as doxorubicin, and in combination with a checkpoint inhibitor. In some embodiments, the additional chemotherapeutic agent is doxorubicin.

[0119] Doxorubicin is a small molecule compound with the formula CAS 23214-92-8 and is sold under the trade name Adriamycin. DOXIL® is the trade name for polyethylene glycol coated liposome-encapsulated doxorubicin.

[0120] In another aspect, the present invention provides a method of determining an effective amount of ketotifen in a subject having a solid tumor, comprising: (a) measuring the blood flow and / or hardness of the solid tumor; (b) administering an effective amount of ketotifen to the subject; and (c) measuring the blood flow and / or hardness of the solid tumor after administration of ketotifen, wherein an increase in blood flow and / or a decrease in hardness after administration of ketotifen to the subject indicates that the amount administered was an effective amount. In another aspect, the present invention provides a method of treating a solid tumor in a subject in need thereof, comprising: (a) measuring the blood flow and / or hardness of the solid tumor; (b) administering an effective amount of ketotifen to the subject; (c) measuring the blood flow and / or hardness of the solid tumor after administration of ketotifen; and (d) administering a chemotherapeutic agent if the blood flow of the solid tumor increases and / or the hardness of the solid tumor decreases after administration of ketotifen. In another aspect, the invention provides a method of treating a solid tumor in a subject in need thereof, comprising: (a) measuring the blood flow and / or hardness of the solid tumor; (b) administering an effective amount of ketotifen to the subject; (c) measuring the blood flow and / or hardness of the solid tumor after administration of ketotifen; (d) determining that the subject is responsive to a chemotherapeutic agent based on an increase in blood flow of the solid tumor or a decrease in hardness of the solid tumor after administration of ketotifen; and (e) administering a chemotherapeutic agent to the subject who has been determined to be responsive to the chemotherapeutic agent based on an increase in blood flow of the solid tumor or a decrease in hardness of the solid tumor after administration of ketotifen. In another aspect, the invention provides a method for predicting response to treatment with a chemotherapeutic agent, comprising: (a) measuring the blood flow and / or stiffness of a solid tumor; (b) administering to a subject an effective amount of ketotifen; and (c) measuring the blood flow and / or stiffness of the solid tumor after administration of ketotifen, wherein an increase in blood flow of the solid tumor or a decrease in stiffness of the solid tumor after administration of ketotifen indicates that the subject is likely to respond to treatment with the chemotherapeutic agent.In some embodiments, the effective amount of ketotifen is determined by measuring a change in blood flow and / or stiffness of the solid tumor following administration of ketotifen to the subject, and an increase in blood flow and / or a decrease in stiffness following administration of ketotifen to the subject indicates that the amount administered was an effective amount. In some embodiments, the method includes measuring blood flow of the solid tumor, and blood flow of the solid tumor increases following administration of ketotifen. In some embodiments, the method includes measuring stiffness of the solid tumor, and stiffness of the solid tumor decreases following administration of ketotifen. In some embodiments, ketotifen is administered at least 1 day, at least 2 days, at least 3 days, at least 4 days, or at least 5 days prior to administration of the chemotherapeutic agent. In some embodiments, ketotifen is administered at a dose that increases blood flow of the solid tumor and / or decreases stiffness of the solid tumor. In some embodiments, blood flow and / or stiffness of the solid tumor is measured using ultrasound. In some embodiments, blood flow of the solid tumor is measured using a histology technique to measure hypoxia. In some embodiments, the chemotherapeutic agent is a checkpoint inhibitor. In some embodiments, the subject is a human.

[0121] In some embodiments of any of the aspects provided herein, administration of ketotifen reduces tissue stiffness of the solid tumor. In some embodiments of any of the aspects provided herein, administration of ketotifen, or a pharma- ceutically acceptable salt thereof, reduces tissue stiffness of the solid tumor. In some embodiments, the tissue stiffness of the solid tumor is reduced by at least 10%. In some embodiments, the tissue stiffness of the solid tumor is reduced by at least 20%. In some embodiments, the tissue stiffness of the solid tumor is reduced by at least 25%. In some embodiments, the tissue stiffness of the solid tumor is reduced by at least 30%. In some embodiments, the tissue stiffness of the solid tumor is reduced by at least 40%. In some embodiments, the tissue stiffness of the solid tumor is reduced by at least 50%. In some embodiments, the tissue stiffness of the solid tumor is reduced by at least 60%. In some embodiments, the tissue stiffness of the solid tumor is reduced by at least 70%. In some embodiments, the tissue stiffness of the solid tumor is reduced by at least 75%. In some embodiments, the tissue stiffness of the solid tumor is measured using ultrasound elastography.

[0122] In some embodiments of any of the aspects provided herein, administering ketotifen reduces the level of the extracellular matrix protein in the solid tumor. In some embodiments of any of the aspects provided herein, administering ketotifen or a pharma- ceutically acceptable salt thereof reduces the level of the extracellular matrix protein in the solid tumor. In some embodiments, the level of the extracellular matrix protein in the solid tumor is reduced by at least 10%. In some embodiments, the level of the extracellular matrix protein in the solid tumor is reduced by at least 20%. In some embodiments, the level of the extracellular matrix protein in the solid tumor is reduced by at least 25%. In some embodiments, the level of the extracellular matrix protein in the solid tumor is reduced by at least 30%. In some embodiments, the level of the extracellular matrix protein in the solid tumor is reduced by at least 40%. In some embodiments, the level of the extracellular matrix protein in the solid tumor is reduced by at least 50%. In some embodiments, the level of the extracellular matrix protein in the solid tumor is reduced by at least 60%. In some embodiments, the level of the extracellular matrix protein in the solid tumor is reduced by at least 70%. In some embodiments, the level of the extracellular matrix protein in the solid tumor is reduced by at least 75%. In some embodiments, the extracellular matrix protein is collagen I. In some embodiments, the extracellular matrix protein is hyaluronan binding protein (HABP).

[0123] In some embodiments of any of the aspects provided herein, administering ketotifen reduces hypoxia in the solid tumor. In some embodiments of any of the aspects provided herein, administering ketotifen, or a pharma- ceutically acceptable salt thereof, reduces hypoxia in the solid tumor. In some embodiments, the hypoxia is reduced by at least 10%. In some embodiments, the hypoxia is reduced by at least 20%. In some embodiments, the hypoxia is reduced by at least 25%. In some embodiments, the hypoxia is reduced by at least 30%. In some embodiments, the hypoxia is reduced by at least 40%. In some embodiments, the hypoxia is reduced by at least 50%. In some embodiments, the hypoxia is reduced by at least 60%. In some embodiments, the hypoxia is reduced by at least 70%. In some embodiments, the hypoxia is reduced by at least 75%.

[0124] In some embodiments of any of the aspects provided herein, the solid tumor is selected from the group consisting of mesothelioma, breast cancer, lung metastasis of breast cancer, sarcoma, pancreatic cancer, ovarian cancer, liver metastasis, prostate cancer, brain cancer, melanoma, renal cell carcinoma, colorectal cancer, hepatocellular carcinoma, lung cancer, head and neck squamous cell carcinoma, urothelial carcinoma, esophageal squamous cell carcinoma, gastric cancer, esophageal cancer, cervical cancer, Merkel cell carcinoma, endometrial carcinoma, and cutaneous squamous cell carcinoma. In some embodiments, the solid tumor is breast cancer. In some embodiments, the breast cancer has high tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the breast cancer is triple negative breast cancer. In some embodiments, the solid tumor is mesothelioma. In some embodiments, the solid tumor is lung metastasis of breast cancer. In some embodiments, the solid tumor is a sarcoma. In some embodiments, the solid tumor is an osteosarcoma. In some embodiments, the solid tumor is a fibrosarcoma. In some embodiments, the solid tumor is pancreatic cancer. In some embodiments, the solid tumor is ovarian cancer. In some embodiments, the solid tumor is a liver metastasis. In some embodiments, the liver metastasis is from colorectal cancer. In some embodiments, the solid tumor is prostate cancer. In some embodiments, the prostate cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the solid tumor is brain cancer. In some embodiments, the brain cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the solid tumor is melanoma. In some embodiments, the solid tumor is renal cell carcinoma. In some embodiments, the solid tumor is colorectal cancer. In some embodiments, the colorectal cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the colorectal cancer has low tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the colorectal cancer has high tumor endothelin-A receptor expression and low endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the solid tumor is hepatocellular carcinoma. In some embodiments, the solid tumor is lung cancer. In some embodiments, the lung cancer expresses endothelin-A receptor. In some embodiments, the lung cancer expresses an endothelin-B receptor.In some embodiments, the lung cancer expresses both endothelin-A and endothelin-B receptors. In some embodiments, the lung cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the lung cancer has high tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the lung cancer has high tumor endothelin-A and endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the lung cancer is non-small cell lung cancer. In some embodiments, the lung cancer is small cell lung cancer. In some embodiments, the solid tumor is head and neck squamous cell carcinoma. In some embodiments, the solid tumor is urothelial carcinoma. In some embodiments, the solid tumor is esophageal squamous cell carcinoma. In some embodiments, the solid tumor is gastric cancer. In some embodiments, the solid tumor is esophageal cancer. In some embodiments, the solid tumor is cervical cancer. In some embodiments, the solid tumor is Merkel cell carcinoma. In some embodiments, the solid tumor is endometrial cancer. In some embodiments, the solid tumor is cutaneous squamous cell carcinoma. In some embodiments, the solid tumor is a compressed vascular and / or hypoperfused cancer. In some embodiments, the solid tumor is a compressed vascular cancer. In some embodiments, the solid tumor is a hypoperfused cancer. In some embodiments, the compressed vascular and / or hypoperfused solid tumor is selected from the group consisting of breast cancer, lung metastasis of breast cancer, pancreatic cancer, ovarian cancer, and liver metastasis. In some embodiments, the compressed vascular and / or hypoperfused solid tumor is breast cancer. In some embodiments, the breast cancer has high tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the breast cancer is triple-negative breast cancer. In some embodiments, the compressed vascular and / or hypoperfused solid tumor is pancreatic cancer. In some embodiments, the compressed vascular and / or hypoperfused solid tumor is ovarian cancer. In some embodiments, the compressed vascular and / or hypoperfused solid tumor is a liver metastasis. In some embodiments, the liver metastasis is from colorectal cancer.In some embodiments, the solid tumor with compressed blood vessels and / or hypoperfusion is a lung metastasis. In some embodiments, the liver metastasis is from breast cancer. In some embodiments, the solid tumor is a cancer with endothelin receptor expression in tumor vasculature and / or fibroblasts. In some embodiments, the solid tumor is a cancer with endothelin receptor expression in tumor vasculature. In some embodiments, the solid tumor is a cancer with endothelin receptor expression in tumor fibroblasts. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is selected from the group consisting of pancreatic cancer, ovarian cancer, lung cancer, prostate cancer, brain cancer, breast cancer, and colorectal cancer. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is pancreatic cancer. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is ovarian cancer. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is lung cancer. In some embodiments, the lung cancer expresses endothelin-A receptors. In some embodiments, the lung cancer expresses endothelin-B receptors. In some embodiments, the lung cancer expresses both endothelin-A and endothelin-B receptors. In some embodiments, the lung cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the lung cancer has high tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the lung cancer has high tumor endothelin-A receptor and endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the lung cancer is non-small cell lung cancer. In some embodiments, the lung cancer is small cell lung cancer. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is prostate cancer. In some embodiments, the prostate cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is brain cancer.In some embodiments, the brain cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is breast cancer. In some embodiments, the breast cancer has high tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the breast cancer is triple-negative breast cancer. In some embodiments, the solid tumor is a lung metastasis from breast cancer. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is colorectal cancer. In some embodiments, the colorectal cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the colorectal cancer has low tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the colorectal cancer has high tumor endothelin-A receptor expression and low endothelin-B receptor expression compared to non-tumor tissue.

[0125] B. Route of Administration The chemotherapeutic agent described herein can be administered by any suitable route and manner. Ketotifen or its pharmaceutically acceptable salt, or the checkpoint inhibitor described herein can be administered by any suitable route and manner. Suitable routes of administration of the compounds or antibodies of the present invention are well known in the art and can be selected by the skilled artisan. In one embodiment, ketotifen or its pharmaceutically acceptable salt, and / or the checkpoint inhibitor described herein is administered parenterally. Parenteral administration refers to a mode of administration other than intestinal administration and topical administration, usually by injection, and includes epidermal, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratendinous, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, intracranial, intrasternal, epidural, and intracisternal injection and infusion. In some embodiments, the administration route of the chemotherapeutic agent is intraperitoneal injection. In some embodiments, the administration route of the chemotherapeutic agent is intravenous injection. In some embodiments, the route of administration of ketotifen or a pharma- ceutically acceptable salt thereof is intraperitoneal injection. In some embodiments, the route of administration of the checkpoint inhibitor is intraperitoneal injection. In some embodiments, the route of administration of ketotifen or a pharma- ceutically acceptable salt thereof is intravenous injection. In some embodiments, the route of administration of the checkpoint inhibitor is intravenous injection. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof, and / or a checkpoint inhibitor described herein is administered enterally. In some embodiments, the route of administration of ketotifen or a pharma- ceutically acceptable salt thereof is enteral. In some embodiments, the route of administration of ketotifen or a pharma- ceutically acceptable salt thereof is oral. In some embodiments, the route of administration of the checkpoint inhibitor is enteral. In some embodiments, the route of administration of the checkpoint inhibitor is oral. In some embodiments, the route of administration of the chemotherapeutic agent is enteral. In some embodiments, the route of administration of the chemotherapeutic agent is oral.

[0126] C. Dose and frequency of administration In one aspect, the invention provides a method as described herein, comprising administering ketotifen or a pharma- ceutically acceptable salt thereof as described herein and a checkpoint inhibitor as described herein, wherein the subject is administered ketotifen or a pharma- ceutically acceptable salt thereof as described herein and a checkpoint inhibitor as described herein at a particular frequency. In another aspect, the invention provides a method as described herein, comprising administering ketotifen as described herein and a chemotherapeutic agent as described herein, wherein the subject is administered ketotifen as described herein and a chemotherapeutic agent as described herein at a particular frequency.

[0127] In one embodiment of the method, or use, or product for use provided herein, ketotifen as described herein is administered to the subject in a therapeutically effective amount. In one embodiment of the method, or use, or product for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered to the subject in a therapeutically effective amount. In one embodiment of the method, or use, or product for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered to the subject in a sub-therapeutic dose. In one embodiment of the method, or use, or product for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered to the subject in a dose sufficient to elicit the effect of the checkpoint inhibitor. In one embodiment of the method, or use, or product for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered to the subject in a dose sufficient to enhance the effect of the checkpoint inhibitor. In one embodiment of the method, or use, or product for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to the subject in a dose sufficient to prolong the effect of the checkpoint inhibitor. In one embodiment of the method, or use, or product for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to the subject in a dose sufficient to enhance the effect of the checkpoint inhibitor. In one embodiment of the method, or use, or product for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to the subject in a dose sufficient to improve the delivery of the checkpoint inhibitor to a solid tumor. In one embodiment of the method, or use, or product for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to the subject in a dose sufficient to improve the efficacy of the checkpoint inhibitor.In one embodiment of the method, or use, or product for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to a subject in a dose sufficient to increase the number of anti-tumor T cells localized to a solid tumor. In one embodiment of the method, or use, or product for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to a subject in a dose sufficient to reduce the tissue hardness of a solid tumor. In one embodiment of the method, or use, or product for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to a subject in a dose sufficient to reduce the level of extracellular matrix proteins in a solid tumor. In one embodiment of the method, or use, or product for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to a subject in a dose sufficient to increase the blood flow of a solid tumor. In one embodiment of the methods or uses or products for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to a subject at a dose sufficient to reduce the level of extracellular matrix proteins in a solid tumor and increase blood flow to the solid tumor. In one embodiment of the methods or uses or products for use provided herein, ketotifen or a pharma-ceutically acceptable salt thereof described herein is administered to a subject at a dose sufficient to reduce hypoxia in a solid tumor.

[0128] In some embodiments of the methods, or uses, or products for use provided herein, ketotifen as described herein is administered to a subject at a dose ranging from about 0.01 mg / kg to about 20 mg / kg of the subject's body weight. In some embodiments of the methods, or uses, or products for use provided herein, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered to a subject at a dose ranging from about 0.01 mg / kg to about 20 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered at a dose ranging from about 0.05 mg / kg to about 15 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered at a dose ranging from about 0.01 mg / kg to about 0.1 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered at a dose ranging from about 0.01 mg / kg to about 0.5 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.01 mg / kg to about 1.0 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.01 mg / kg to about 5 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.05 mg / kg to about 0.1 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.05 mg / kg to about 10 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.05 mg / kg to about 5 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.05 mg / kg to about 3 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.25 mg / kg to about 10 mg / kg of the subject's body weight.In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.25 mg / kg to about 5 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.25 mg / kg to about 3 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.5 mg / kg to about 10 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.5 mg / kg to about 5 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.5 mg / kg to about 3 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.75 mg / kg to about 10 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.75 mg / kg to about 5 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 0.75 mg / kg to about 3 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 1 mg / kg to about 10 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 1 mg / kg to about 5.0 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 1 mg / kg to about 3 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 2 mg / kg to about 20 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 2 mg / kg to about 15 mg / kg of subject body weight.In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 2 mg / kg to about 10 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 2 mg / kg to about 5 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 4 mg / kg to about 20 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 4 mg / kg to about 15 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 4 mg / kg to about 10 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 4 mg / kg to about 5 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 5 mg / kg to about 20 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.01 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.05 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.1 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.15 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.16 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.2 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.3 mg / kg of the subject's body weight.In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.4 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.5 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.6 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.7 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.8 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 0.9 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 1 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 1.2 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 1.4 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 1.6 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 1.8 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 2 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 2.2 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 2.4 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 2.6 mg / kg of the subject's body weight.In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 2.8 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 3 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 3.2 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 3.4 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 3.6 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 3.8 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 4 mg / kg of subject body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 4.2 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 4.4 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 4.6 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 4.8 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 5 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 5.2 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 5.4 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 5.6 mg / kg of the subject's body weight.In one embodiment, ketotifen or a pharmaceutically acceptable salt thereof described herein is administered at a dose of about 5.8 mg / kg of subject body weight. In one embodiment, ketotifen or a pharmaceutically acceptable salt thereof described herein is administered at a dose of about 6 mg / kg of subject body weight. In one embodiment, ketotifen or a pharmaceutically acceptable salt thereof described herein is administered at a dose of about 6.5 mg / kg of subject body weight. In one embodiment, ketotifen or a pharmaceutically acceptable salt thereof described herein is administered at a dose of about 7 ... In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 7.5 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 8 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 8.5 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 9 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 9.5 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 10 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 11 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 12 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 13 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 14 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 15 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 16 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 17 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 18 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 19 mg / kg of the subject's body weight. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 20 mg / kg of the subject's body weight.

[0129] In some embodiments of the methods or uses or products for use provided herein, ketotifen as described herein is administered to a subject at a dose ranging from about 10 mg to about 1250 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered at a dose ranging from about 10 mg to about 150 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered at a dose ranging from about 10 mg to about 100 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered at a dose ranging from about 10 mg to about 50 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered at a dose ranging from about 25 mg to about 150 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered at a dose ranging from about 25 mg to about 100 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose range of about 25 mg to about 50 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose range of about 50 mg to about 150 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose range of about 50 mg to about 100 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose range of about 50 mg to about 75 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose range of about 75 mg to about 150 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose range of about 75 mg to about 100 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 100 mg to about 1200 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose ranging from about 10 mg to about 40 mg.In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose ranging from about 10 mg to about 30 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose ranging from about 10 mg to about 20 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose ranging from about 15 mg to about 40 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose ranging from about 20 mg to about 40 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose ranging from about 30 mg to about 40 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 10 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 15 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 20 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 25 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 30 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 35 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 40 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 45 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 50 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 55 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 60 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 62.5 mg.In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 65 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 70 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 75 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 80 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 85 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 90 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 95 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 100 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 105 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 110 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 115 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 120 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 125 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 130 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 135 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 140 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 145 mg.In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 150 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 175 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 200 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 250 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 300 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 350 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 400 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 450 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 500 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 550 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 600 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 650 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 700 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered in a dose of about 750 mg. In one embodiment, ketotifen or its pharmaceutically acceptable salt as described herein is administered at a dose of about 800 mg. In one embodiment, ketotifen or its pharmaceutically acceptable salt as described herein is administered at a dose of about 850 mg. In one embodiment, ketotifen or its pharmaceutically acceptable salt as described herein is administered at a dose of about 900 mg.In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 950 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 1000 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 1050 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 1100 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 1150 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 1200 mg. In one embodiment, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered at a dose of about 1250 mg.

[0130] In one embodiment of the method or use or product for use provided herein, ketotifen is administered to the subject daily, twice a day, three times a day, or four times a day. In some embodiments, ketotifen or a pharmaceutically acceptable salt thereof is administered to the subject once every two days, about once a week, or about once every three weeks. In some embodiments, ketotifen or a pharmaceutically acceptable salt thereof is administered to the subject about once a day. In some embodiments, ketotifen or a pharmaceutically acceptable salt thereof is administered to the subject about twice a day. In some embodiments, ketotifen or a pharmaceutically acceptable salt thereof is administered to the subject once a day. In some embodiments, ketotifen or a pharmaceutically acceptable salt thereof is administered to the subject twice a day. In some embodiments, ketotifen or a pharmaceutically acceptable salt thereof is administered to the subject orally.

[0131] In some embodiments of the methods, or uses, or products for use provided herein, the chemotherapeutic agent described herein is administered to the subject at a dose ranging from about 0.5 mg / kg to about 15 mg / kg of subject body weight. In some embodiments of the methods, or uses, or products for use provided herein, the checkpoint inhibitor described herein is administered to the subject at a dose ranging from about 0.5 mg / kg to about 15 mg / kg of subject body weight. In one embodiment, the checkpoint inhibitor described herein is administered at a dose ranging from about 1 mg / kg to about 10 mg / kg. In one embodiment, the checkpoint inhibitor described herein is administered at a dose of about 1 mg / kg of subject body weight. In one embodiment, the checkpoint inhibitor described herein is administered at a dose of about 2 mg / kg of subject body weight. In one embodiment, the checkpoint inhibitor described herein is administered at a dose of about 3 mg / kg of subject body weight. In one embodiment, the checkpoint inhibitor described herein is administered at a dose of about 4 mg / kg of subject body weight. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 5 mg / kg of the subject's body weight. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 6 mg / kg of the subject's body weight. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 7 mg / kg of the subject's body weight. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 8 mg / kg of the subject's body weight. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 9 mg / kg of the subject's body weight. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 10 mg / kg of the subject's body weight. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 11 mg / kg of the subject's body weight. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 12 mg / kg of the subject's body weight. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 13 mg / kg of the subject's body weight. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 14 mg / kg of the subject's body weight.In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 15 mg / kg of subject body weight. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 2 mg / kg, and the checkpoint inhibitor is pembrolizumab. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 1 mg / kg, and the checkpoint inhibitor is nivolumab. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 3 mg / kg, and the checkpoint inhibitor is nivolumab. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 1 mg / kg, and the checkpoint inhibitor is ipilimumab. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 3 mg / kg, and the checkpoint inhibitor is ipilimumab. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 10 mg / kg, and the checkpoint inhibitor is ipilimumab.

[0132] In some embodiments of the methods, or uses, or products for use provided herein, the chemotherapeutic agent described herein is administered to the subject at a dose ranging from about 100 mg to about 2000 mg. In some embodiments of the methods, or uses, or products for use provided herein, the checkpoint inhibitor described herein is administered to the subject at a dose ranging from about 100 mg to about 2000 mg. In one embodiment, the checkpoint inhibitor described herein is administered at a dose ranging from about 200 mg to about 1800 mg. In one embodiment, the checkpoint inhibitor described herein is administered at a dose ranging from about 200 mg to about 400 mg. In one embodiment, the checkpoint inhibitor described herein is administered at a dose ranging from about 400 mg to about 600 mg. In one embodiment, the checkpoint inhibitor described herein is administered at a dose ranging from about 600 mg to about 1000 mg. In one embodiment, the checkpoint inhibitor described herein is administered at a dose ranging from about 800 mg to about 1000 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose range of about 1000 mg to about 1800 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose range of about 1000 mg to about 1600 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose range of about 1000 mg to about 1300 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose range of about 140 mg to about 1800 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose range of about 1600 mg to about 1800 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose range of about 100 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose range of about 200 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 240 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 300 mg. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 360 mg.In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 400 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 480 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 500 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 600 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 700 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 800 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 840 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 900 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 1000 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 1100 mg. In one embodiment, the checkpoint inhibitors described herein are administered in a dose of about 1200 mg. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 1300 mg. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 1400 mg. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 1500 mg. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 1600 mg. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 1700 mg. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 1800 mg. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 1900 mg. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 2000 mg. In one embodiment, the checkpoint inhibitors described herein are administered at a dose of about 200 mg, and the checkpoint inhibitor is pembrolizumab.In one embodiment, the checkpoint inhibitor described herein is administered at a dose of about 400 mg, and the checkpoint inhibitor is pembrolizumab. In one embodiment, the checkpoint inhibitor described herein is administered at a dose of about 240 mg, and the checkpoint inhibitor is nivolumab. In one embodiment, the checkpoint inhibitor described herein is administered at a dose of about 480 mg, and the checkpoint inhibitor is nivolumab. In one embodiment, the checkpoint inhibitor described herein is administered at a dose of about 360 mg, and the checkpoint inhibitor is nivolumab. In one embodiment, the checkpoint inhibitor described herein is administered at a dose of about 840 mg, and the checkpoint inhibitor is atezolizumab. In one embodiment, the checkpoint inhibitor described herein is administered at a dose of about 1200 mg, and the checkpoint inhibitor is atezolizumab. In one embodiment, the checkpoint inhibitor described herein is administered at a dose of about 1680 mg, and the checkpoint inhibitor is atezolizumab.

[0133] In one embodiment of the methods, or uses, or products for use provided herein, the chemotherapeutic agents described herein are administered to the subject daily, twice daily, three times daily, or four times daily. In one embodiment of the methods, or uses, or products for use provided herein, the checkpoint inhibitors described herein are administered to the subject daily, twice daily, three times daily, or four times daily. In some embodiments, the checkpoint inhibitors described herein are administered about once every week to about once every eight weeks. In some embodiments, the checkpoint inhibitors described herein are administered about once every week. In some embodiments, the checkpoint inhibitors described herein are administered about once every two weeks. In some embodiments, the checkpoint inhibitors described herein are administered about once every three weeks. In some embodiments, the checkpoint inhibitors described herein are administered about once every four weeks. In some embodiments, the checkpoint inhibitors described herein are administered about once every five weeks. In some embodiments, the checkpoint inhibitors described herein are administered about once every six weeks. In some embodiments, the checkpoint inhibitors described herein are administered about once every 7 weeks. In some embodiments, the checkpoint inhibitors described herein are administered about once every 8 weeks. In some embodiments, the checkpoint inhibitors described herein are administered about once every 3 weeks, and the checkpoint inhibitor is pembrolizumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 200 mg once every 3 weeks, and the checkpoint inhibitor is pembrolizumab. In some embodiments, the checkpoint inhibitors described herein are administered about once every 6 weeks, and the checkpoint inhibitor is pembrolizumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 400 mg once every 6 weeks, and the checkpoint inhibitor is pembrolizumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 2 mg / kg of subject body weight once every 3 weeks, and the checkpoint inhibitor is pembrolizumab.In some embodiments, the checkpoint inhibitors described herein are administered about once every two weeks, and the checkpoint inhibitor is nivolumab. In some embodiments, the checkpoint inhibitors described herein are administered about once every two weeks at a dose of about 240 mg, and the checkpoint inhibitor is nivolumab. In some embodiments, the checkpoint inhibitors described herein are administered about once every three weeks, and the checkpoint inhibitor is nivolumab. In some embodiments, the checkpoint inhibitors described herein are administered about once every three weeks at a dose of about 360 mg, and the checkpoint inhibitor is nivolumab. In some embodiments, the checkpoint inhibitors described herein are administered about once every four weeks, and the checkpoint inhibitor is nivolumab. In some embodiments, the checkpoint inhibitors described herein are administered about once every four weeks at a dose of about 480 mg, and the checkpoint inhibitor is nivolumab. In some embodiments, the checkpoint inhibitors described herein are administered about once every three weeks at a dose of about 1 mg / kg, and the checkpoint inhibitor is nivolumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 3 mg / kg about once every two weeks, and the checkpoint inhibitor is nivolumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 3 mg / kg about once every three weeks, and the checkpoint inhibitor is nivolumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 3 mg / kg about once every three weeks, and the checkpoint inhibitor is ipilimumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 1 mg / kg about once every three weeks, and the checkpoint inhibitor is ipilimumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 3 mg / kg about once every three weeks, and the checkpoint inhibitor is ipilimumab.In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 10 mg / kg about once every 3 weeks, and the checkpoint inhibitor is ipilimumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 10 mg / kg about once every 12 weeks, and the checkpoint inhibitor is ipilimumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 1 mg / kg about once every 6 weeks, and the checkpoint inhibitor is ipilimumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 2 weeks, and the checkpoint inhibitor is atezolizumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 840 mg about once every 2 weeks, and the checkpoint inhibitor is atezolizumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 3 weeks, and the checkpoint inhibitor is atezolizumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 1200 mg once every about three weeks, and the checkpoint inhibitor is atezolizumab. In some embodiments, the checkpoint inhibitors described herein are administered at a dose of about 1680 mg once every about four weeks, and the checkpoint inhibitor is atezolizumab. In some embodiments, the checkpoint inhibitors described herein are administered to the subject by intravenous infusion.

[0134] D. Treatment outcomes In one aspect, the method of treating cancer with ketotifen described herein and the chemotherapeutic agents described herein results in an improvement in one or more therapeutic effects in the subject after administration compared to baseline. In one aspect, the method of treating cancer with ketotifen or a pharma- ceutically acceptable salt thereof described herein and the checkpoint inhibitors described herein results in an improvement in one or more therapeutic effects in the subject after administration compared to baseline. In some embodiments, the one or more therapeutic effects are the size of a tumor derived from a cancer (e.g., a solid tumor), an objective response rate, duration of response, time to response, progression-free survival, overall survival, or any combination thereof. In one embodiment, the one or more therapeutic effects are the size of a tumor derived from a cancer. In one embodiment, the one or more therapeutic effects are a reduction in tumor size. In one embodiment, the one or more therapeutic effects are stable disease. In one embodiment, the one or more therapeutic effects are a partial response. In one embodiment, the one or more therapeutic effects are a complete response. In one embodiment, the one or more therapeutic effects are an objective response rate. In one embodiment, the one or more therapeutic effects are duration of response. In one embodiment, the one or more therapeutic effects are time to response. In one embodiment, the one or more therapeutic benefits is progression-free survival. In one embodiment, the one or more therapeutic benefits is overall survival. In one embodiment, the one or more therapeutic benefits is regression of the cancer.

[0135] In one embodiment of the method or use or product for use provided herein, the response to treatment with ketotifen as described herein and chemotherapeutic agents as described herein may include RECIST Criteria 1.1. In one embodiment of the method or use or product for use provided herein, the response to treatment with ketotifen or its pharma- ceutically acceptable salts as described herein and checkpoint inhibitors as described herein may include RECIST Criteria 1.1. RECIST Criteria 1.1 is as follows:

Table 4

[0136] In one embodiment of the methods, or uses, or products for use provided herein, the efficacy of treatment with ketotifen described herein and chemotherapeutic agents described herein is assessed by measuring the objective response rate. In one embodiment of the methods, or uses, or products for use provided herein, the efficacy of treatment with ketotifen or a pharma- ceutically acceptable salt thereof described herein and a checkpoint inhibitor described herein is assessed by measuring the objective response rate. In some embodiments, the objective response rate is the proportion of patients in a minimum period of time who have a predefined amount of tumor size reduction. In some embodiments, the objective response rate is based on RECIST v1.1. In one embodiment, the objective response rate is at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, or at least about 80%. In one embodiment, the objective response rate is at least about 20%-80%. In one embodiment, the objective response rate is at least about 30%-80%. In one embodiment, the objective response rate is at least about 40%-80%. In one embodiment, the objective response rate is at least about 50%-80%. In one embodiment, the objective response rate is at least about 60%-80%. In one embodiment, the objective response rate is at least about 70%-80%. In one embodiment, the objective response rate is at least about 80%. In one embodiment, the objective response rate is at least about 85%. In one embodiment, the objective response rate is at least about 90%. In one embodiment, the objective response rate is at least about 95%. In one embodiment, the objective response rate is at least about 98%. In one embodiment, the objective response rate is at least about 99%. In one embodiment, the objective response rate is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, or at least 80%. In one embodiment, the objective response rate is at least 20%-80%. In one embodiment, the objective response rate is at least 30% to 80%.In one embodiment, the objective response rate is at least 40%-80%. In one embodiment, the objective response rate is at least 50%-80%. In one embodiment, the objective response rate is at least 60%-80%. In one embodiment, the objective response rate is at least 70%-80%. In one embodiment, the objective response rate is at least 80%. In one embodiment, the objective response rate is at least 85%. In one embodiment, the objective response rate is at least 90%. In one embodiment, the objective response rate is at least 95%. In one embodiment, the objective response rate is at least 98%. In one embodiment, the objective response rate is at least 99%. In one embodiment, the objective response rate is 100%.

[0137] In one embodiment of the methods, or uses, or products for use provided herein, the response to treatment with ketotifen described herein and chemotherapeutic agents described herein is assessed by measuring the size of a tumor derived from a cancer (e.g., a solid tumor). In one embodiment of the methods, or uses, or products for use provided herein, the response to treatment with ketotifen or a pharma- ceutically acceptable salt thereof described herein and a checkpoint inhibitor described herein is assessed by measuring the size of a tumor derived from a cancer (e.g., a solid tumor). In one embodiment, the size of a tumor derived from a cancer is reduced by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, or at least about 80% relative to the size of a tumor derived from a cancer prior to administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein and / or a checkpoint inhibitor described herein. In one embodiment, the size of a tumor derived from a cancer is reduced by at least about 10% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least about 20% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least about 30% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least about 40% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least about 50% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least about 60% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least about 70% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least about 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least about 85%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least about 90%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least about 95%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least about 98%.In one embodiment, the size of the tumor derived from the cancer is reduced by at least about 99%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, or at least 80% relative to the size of the tumor derived from the cancer prior to administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein and / or a checkpoint inhibitor described herein. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 10% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 20% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 30% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 40% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 50% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 60% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 70% to 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 80%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 85%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 90%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 95%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 98%. In one embodiment, the size of the tumor derived from the cancer is reduced by at least 99%. In one embodiment, the size of the tumor derived from the cancer is reduced by 100%. In one embodiment, the size of the tumor derived from the cancer is measured by magnetic resonance imaging (MRI). In one embodiment, the size of the tumor derived from the cancer is measured by computed tomography (CT). In some embodiments, the size of the tumor derived from the cancer is reduced compared to the size of the tumor before administration of ketotifen or a pharma- ceutically acceptable salt thereof as described herein and a checkpoint inhibitor as described herein.In some embodiments, the size of the tumor derived from the cancer is reduced compared to the size of the tumor before administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein. In some embodiments, the size of the tumor derived from the cancer is reduced compared to the size of the tumor before administration of a checkpoint inhibitor described herein.

[0138] In one embodiment of the method or use or product for use described herein, the response to treatment with ketotifen as described herein and chemotherapeutic agents as described herein is assessed by measuring progression-free survival after administration of ketotifen as described herein and / or chemotherapeutic agents as described herein.In one embodiment of the method or use or product for use described herein, the response to treatment with ketotifen or a pharmaceutically acceptable salt thereof as described herein and a checkpoint inhibitor as described herein is assessed by measuring progression-free survival after administration of ketotifen or a pharmaceutically acceptable salt thereof as described herein and / or a checkpoint inhibitor as described herein. In some embodiments, the subject exhibits a progression-free survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration of ketotifen or a pharmaceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits a progression-free survival of at least about 6 months after administration of ketotifen or a pharmaceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits a progression-free survival of at least about 1 year after administration of ketotifen or a pharmaceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits a progression-free survival of at least about 2 years following administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein.In some embodiments, the subject exhibits a progression-free survival of at least about 3 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits a progression-free survival of at least about 4 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits a progression-free survival of at least about 5 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits a progression-free survival of at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 2 years, at least 3 years, at least 4 years, or at least 5 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits a progression-free survival of at least 6 months after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits a progression-free survival of at least 1 year after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits a progression-free survival of at least 2 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits progression-free survival of at least 3 years following administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein.In some embodiments, the subject exhibits a progression-free survival of at least 4 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits a progression-free survival of at least 5 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the response to treatment is assessed by measuring the progression-free survival after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein and a checkpoint inhibitor described herein. In some embodiments, the response to treatment is assessed by measuring the progression-free survival after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein. In some embodiments, the response to treatment is assessed by measuring the progression-free survival after administration of a checkpoint inhibitor described herein.

[0139] In one embodiment of the method or use or product for use described herein, the response to treatment with ketotifen as described herein and chemotherapeutic agents as described herein is assessed by measuring overall survival after administration of ketotifen as described herein and / or chemotherapeutic agents as described herein.In one embodiment of the method or use or product for use described herein, the response to treatment with ketotifen or a pharmaceutically acceptable salt thereof as described herein and a checkpoint inhibitor as described herein is assessed by measuring overall survival after administration of ketotifen or a pharmaceutically acceptable salt thereof as described herein and / or a checkpoint inhibitor as described herein. In some embodiments, the subject exhibits an overall survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits an overall survival of at least about 6 months after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits an overall survival of at least about 1 year after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits an overall survival of at least about 2 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits an overall survival of at least about 3 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein.In some embodiments, the subject exhibits an overall survival of at least about 4 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits an overall survival of at least about 5 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits an overall survival of at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least about 12 months, at least 18 months, at least 2 years, at least 3 years, at least 4 years, or at least 5 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits an overall survival of at least 6 months after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits an overall survival of at least 1 year after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits an overall survival of at least 2 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits an overall survival of at least 3 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the subject exhibits an overall survival of at least 4 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein.In some embodiments, the subject exhibits an overall survival of at least 5 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein, and / or a checkpoint inhibitor described herein. In some embodiments, the response to treatment is assessed by measuring the time of overall survival after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein and a checkpoint inhibitor described herein. In some embodiments, the response to treatment is assessed by measuring the time of overall survival after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein. In some embodiments, the response to treatment is assessed by measuring the time of overall survival after administration of a checkpoint inhibitor described herein.

[0140] In one embodiment of the methods, or uses, or products for use described herein, the response to treatment with ketotifen described herein and chemotherapeutic agents described herein is assessed by measuring the duration of response to ketotifen described herein and chemotherapeutic agents described herein after administration of ketotifen described herein and / or chemotherapeutic agents described herein. In one embodiment of the methods, or uses, or products for use described herein, the response to treatment with ketotifen or a pharma- ceutically acceptable salt thereof described herein and a checkpoint inhibitor described herein is assessed by measuring the duration of response to ketotifen or a pharma- ceutically acceptable salt thereof described herein after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein. The duration of response to ketotifen or its pharmacologic acceptable salts described herein and checkpoint inhibitors described herein after administration of a checkpoint inhibitor is evaluated by measuring. In some embodiments, the duration of response to ketotifen or its pharmacologic acceptable salts described herein and checkpoint inhibitors described herein is at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration of ketotifen or its pharmacologic acceptable salts described herein and / or checkpoint inhibitors described herein. In some embodiments, the duration of response to ketotifen or its pharma- ceutically acceptable salts described herein and checkpoint inhibitors described herein is at least about 6 months after administration of ketotifen or its pharma- ceutically acceptable salts described herein and / or checkpoint inhibitors described herein. In some embodiments, the duration of response to ketotifen or its pharma- ceutically acceptable salts described herein and checkpoint inhibitors described herein is at least about 1 year after administration of ketotifen or its pharma- ceutically acceptable salts described herein and / or checkpoint inhibitors described herein. In some embodiments, the duration of response to ketotifen or its pharma- ceutically acceptable salts described herein and checkpoint inhibitors described herein is at least about 2 years after administration of ketotifen or its pharma- ceutically acceptable salts described herein and / or checkpoint inhibitors described herein.In some embodiments, the duration of response to ketotifen or its pharma- ceutically acceptable salts described herein and the checkpoint inhibitors described herein is at least about 3 years after administration of ketotifen or its pharma- ceutically acceptable salts described herein, and / or the checkpoint inhibitors described herein. In some embodiments, the duration of response to ketotifen or its pharma- ceutically acceptable salts described herein and the checkpoint inhibitors described herein is at least about 4 years after administration of ketotifen or its pharma- ceutically acceptable salts described herein, and / or the checkpoint inhibitors described herein. In some embodiments, the duration of response to ketotifen or its pharma- ceutically acceptable salts described herein and the checkpoint inhibitors described herein is at least about 5 years after administration of ketotifen or its pharma- ceutically acceptable salts described herein, and / or the checkpoint inhibitors described herein. In some embodiments, the duration of response to ketotifen or a pharma- ceutically acceptable salt thereof described herein and the checkpoint inhibitors described herein is at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 2 years, at least 3 years, at least 4 years, or at least 5 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein and / or a checkpoint inhibitor described herein. In some embodiments, the duration of response to ketotifen or a pharma- ceutically acceptable salt thereof described herein and the checkpoint inhibitors described herein is at least 6 months after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein and / or a checkpoint inhibitor described herein.In some embodiments, the duration of response to ketotifen or its pharma- ceutically acceptable salts described herein and checkpoint inhibitors described herein is at least 1 year after administration of ketotifen or its pharma- ceutically acceptable salts described herein and / or checkpoint inhibitors described herein. In some embodiments, the duration of response to ketotifen or its pharma- ceutically acceptable salts described herein and checkpoint inhibitors described herein is at least 2 years after administration of ketotifen or its pharma- ceutically acceptable salts described herein and / or checkpoint inhibitors described herein. In some embodiments, the duration of response to ketotifen or its pharma- ceutically acceptable salts described herein and checkpoint inhibitors described herein is at least 3 years after administration of ketotifen or its pharma- ceutically acceptable salts described herein and / or checkpoint inhibitors described herein. In some embodiments, the duration of response to ketotifen or a pharma- ceutically acceptable salt thereof described herein and a checkpoint inhibitor described herein is at least 4 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein and / or a checkpoint inhibitor described herein. In some embodiments, the duration of response to ketotifen or a pharma- ceutically acceptable salt thereof described herein and a checkpoint inhibitor described herein is at least 5 years after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein and / or a checkpoint inhibitor described herein. In some embodiments, the duration of response is measured after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein and a checkpoint inhibitor described herein. In some embodiments, the duration of response is measured after administration of ketotifen or a pharma- ceutically acceptable salt thereof described herein. In some embodiments, the duration of response is measured after administration of a checkpoint inhibitor described herein.

[0141] V. Composition In some aspects, also provided herein are compositions (e.g., pharmaceutical compositions and therapeutic formulations) comprising ketotifen as described herein and / or a chemotherapeutic agent as described herein. Also provided herein in some aspects are compositions (e.g., pharmaceutical compositions and therapeutic formulations) comprising ketotifen or a pharma- ceutically acceptable salt thereof as described herein, and / or a checkpoint inhibitor as described herein.

[0142] Therapeutic formulations are prepared for storage by mixing the active ingredient having the desired degree of purity with pharma- ceutically acceptable carriers, excipients, or stabilizers as needed (Remington: The Science and Practice of Pharmacy, 20th Ed., Lippincott Williams & Wiklins, Pub., Gennaro Ed., Philadelphia, Pa. 2000).

[0143] Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed and include buffers, antioxidants, such as ascorbic acid, methionine, vitamin E, sodium metabisulfite; preservatives, isotonicity agents, stabilizers, metal complexes (e.g., Zn-protein complexes); chelating agents such as EDTA and / or non-ionic surfactants.

[0144] Buffers can be used to control the pH to a range that optimizes therapeutic efficacy, especially when stability is pH-dependent. Buffers can be present at concentrations ranging from about 50 mM to about 250 mM. Buffers suitable for use with the present invention include both organic and inorganic acids and their salts. For example, citrate, phosphate, succinate, tartrate, fumarate, gluconate, oxalate, lactate, acetate. In addition, buffers can be composed of histidine and trimethylamine salts, such as Tris.

[0145] Preservatives can be added to prevent microbial growth, and are typically present in the range of about 0.2% to 1.0% (weight / volume). Suitable preservatives for use with the present invention include octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium halides (e.g., chloride, bromide, iodide), benzethonium chloride, thimerosal, phenol, butyl, or benzyl alcohol, alkyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol.

[0146] Isotonicity agents, sometimes known as "stabilizers", may be present to adjust or maintain the isotonicity of the liquid in the composition. When used with large charged biomolecules, such as proteins and antibodies, they are often referred to as "stabilizers" because they can interact with the charged groups of amino acid side chains, thereby reducing the potential for inter- and intra-molecular interactions. Isotonicity agents may be present in any amount between about 0.1% and about 25% by weight, or between about 1% and about 5% by weight, taking into account the relative amounts of other components. In some embodiments, isotonicity agents include polyhydric sugar alcohols, trihydric alcohols or higher sugar alcohols, such as glycerin, erythritol, arabitol, xylitol, sorbitol, and mannitol.

[0147] Additional excipients include agents that may act as one or more of the following: (1) bulking agents, (2) dissolution enhancers, (3) stabilizers, and (4) agents that prevent denaturation or adhesion to the walls of the container. Such excipients include polyhydric sugar alcohols (as listed above); amino acids, such as alanine, glycine, glutamine, asparagine, histidine, arginine, lysine, ornithine, leucine, 2-phenylalanine, glutamic acid, threonine, and the like; organic sugars or sugar alcohols, such as sucrose, lactose, lactitol, trehalose, stachyose, mannose, sorbose, xylose, ribose, ribitol, myoinisitose, myoinositol, galactose, galactitol, glycerol, cyclitols (e.g., inositol), polyethylene glycol, sulfur-containing reducing agents such as urea, glutathione, thioctic acid, sodium thioglycolate, thioglycerol, a-monothioglycerol, and sodium thiosulfate; low molecular weight proteins such as human serum albumin, bovine serum albumin, gelatin, or other immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; monosaccharides (e.g., xylose, mannose, fructose, glucose; disaccharides (e.g., lactose, maltose, sucrose); trisaccharides such as raffinose; and polysaccharides such as dextrin or dextran.

[0148] A non-ionic surfactant or detergent (also known as a "wetting agent") may be present to help solubilize the therapeutic agent as well as to protect the therapeutic protein against agitation-induced aggregation, allowing the formulation to be exposed to shear surface stresses without causing denaturation of the active therapeutic protein or antibody. The non-ionic surfactant is present in a range of about 0.05 mg / ml to about 1.0 mg / ml, or about 0.07 mg / ml to about 0.2 mg / ml. In some embodiments, the non-ionic surfactant is present in a range of about 0.001% to about 0.1% w / v, or about 0.01% to about 0.1% w / v, or about 0.01% to about 0.025% w / v.

[0149] Suitable nonionic surfactants include polysorbates (20, 40, 60, 65, 80, etc.), poloxamers (184, 188, etc.), PLURONIC® polyols, TRITON®, polyoxyethylene sorbitan monoethers (TWEEN®-20, TWEEN®-80, etc.), lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50, and 60, glycerol monostearate, sucrose fatty acid esters, methylcellulose, and carboxymethylcellulose. Anionic surfactants that can be used include sodium lauryl sulfate, dioctyl sodium sulfosuccinate, and dioctyl sodium sulfonate. Cationic surfactants include benzalkonium chloride or benzethonium chloride.

[0150] In some embodiments, the formulation comprising ketotifen comprises ketotifen fumarate. In some embodiments, the formulation comprising ketotifen comprises ketotifen fumarate dissolved in ddH2O. In some embodiments, ketotifen is prepared as a liquid formulation. In some embodiments, ketotifen is prepared as an ophthalmic solution. In some embodiments, ketotifen is prepared as a solid, such as a tablet.

[0151] For preparations to be used for in vivo administration, they must be sterile.Preparation can be sterilized by filtration through sterile filtration membrane.The therapeutic composition herein is usually placed in a container with a sterile access port, for example, an intravenous infusion bag or vial with a stopper that can be pierced by a hypodermic injection needle.

[0152] Routes of administration are in accordance with known and accepted methods, for example, by single or multiple boluses or infusion over time in a suitable manner, such as injection or infusion by subcutaneous, intravenous, intraperitoneal, intramuscular, intraarterial, intralesional, or intraarticular routes, topical administration, inhalation, or by sustained or sustained release means.

[0153] The formulation herein may also contain more than one active compound as necessary for the specific indication to be treated, preferably compounds with complementary activities that do not adversely affect each other.Alternatively or in addition, the composition may contain cytotoxic agents, cytokines, or growth inhibitors.Such molecules are suitably present in combination in an amount that is effective for the intended purpose.

[0154] In some embodiments, a composition comprising ketotifen as described herein is co-administered with a composition comprising a chemotherapeutic agent as described herein. In some embodiments, a composition comprising ketotifen or a pharma- ceutically acceptable salt thereof as described herein is co-administered with a composition comprising a checkpoint inhibitor as described herein. In some embodiments, the co-administration is simultaneous or sequential. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof as described herein is administered simultaneously with a checkpoint inhibitor as described herein. In some embodiments, simultaneous means that ketotifen or a pharma- ceutically acceptable salt thereof as described herein and a checkpoint inhibitor as described herein are administered to a subject less than about 1 hour apart, for example less than about 30 minutes apart, less than about 15 minutes apart, less than about 10 minutes apart, or less than about 5 minutes apart. In some embodiments, simultaneous means that ketotifen or a pharma- ceutically acceptable salt thereof as described herein and a checkpoint inhibitor as described herein are administered to a subject less than 1 hour apart, for example less than 30 minutes apart, less than 15 minutes apart, less than 10 minutes apart, or less than 5 minutes apart. In some embodiments, ketotifen, or a pharma- ceutically acceptable salt thereof described herein, is administered sequentially with a checkpoint inhibitor described herein.In some embodiments, sequential administration means that the ketotifen or pharma- ceutically acceptable salts thereof described herein and the checkpoint inhibitors described herein are administered at least 1 hour apart, at least 2 hours apart, at least 3 hours apart, at least 4 hours apart, at least 5 hours apart, at least 6 hours apart, at least 7 hours apart, at least 8 hours apart, at least 9 hours apart, at least 10 hours apart, at least 11 hours apart, at least 12 hours apart, at least 13 hours apart, at least 14 hours apart, at least It means that the two or more are administered at least 15 hours apart, at least 16 hours apart, at least 17 hours apart, at least 18 hours apart, at least 19 hours apart, at least 20 hours apart, at least 21 hours apart, at least 22 hours apart, at least 23 hours apart, at least 24 hours apart, at least 2 days apart, at least 3 days apart, at least 4 days apart, at least 5 days apart, at least 5 days apart, at least 7 days apart, at least 2 weeks apart, at least 3 weeks apart, or at least 4 weeks apart. In some embodiments, ketotifen or a pharmaceutically acceptable salt thereof described herein is administered prior to administration of a checkpoint inhibitor described herein. In some embodiments, ketotifen or a pharmaceutically acceptable salt thereof described herein is administered to a subject beginning at least 1 day prior to administration of a checkpoint inhibitor described herein to the subject. In some embodiments, ketotifen or a pharmaceutically acceptable salt thereof described herein is administered to a subject beginning at least 2 days prior to administration of a checkpoint inhibitor described herein to the subject. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to a subject beginning at least 3 days prior to administering a checkpoint inhibitor described herein to the subject. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to a subject beginning at least 4 days prior to administering a checkpoint inhibitor described herein to the subject.In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to a subject beginning at least 5 days before a checkpoint inhibitor described herein is administered to the subject. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to a subject beginning at least 1 week before a checkpoint inhibitor described herein is administered to the subject. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to a subject beginning at least 2 weeks before a checkpoint inhibitor described herein is administered to the subject. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to a subject beginning at least 3 weeks before a checkpoint inhibitor described herein is administered to the subject. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to a subject beginning at least 4 weeks before a checkpoint inhibitor described herein is administered to the subject. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to the subject beginning before a checkpoint inhibitor described herein is administered to the subject, and administration is maintained for at least a portion of the time the subject is administered the checkpoint inhibitor. In some embodiments, ketotifen or a pharma- ceutically acceptable salt thereof described herein is administered to the subject beginning before a checkpoint inhibitor described herein is administered to the subject, and administration is maintained for the entire time the subject is administered the checkpoint inhibitor.

[0155] In some embodiments, the composition comprising ketotifen or its pharma- ceutically acceptable salts described herein and / or the checkpoint inhibitors described herein is co-administered with one or more additional therapeutic agents. In some embodiments, the co-administration is simultaneous or sequential.

[0156] VI. Products and Kits In another aspect, a product or kit is provided that includes ketotifen as described herein and / or a chemotherapeutic agent as described herein. In another aspect, a product or kit is provided that includes ketotifen or a pharmaceutically acceptable salt thereof as described herein, and / or a checkpoint inhibitor as described herein. The product or kit may further include instructions for using ketotifen or a pharmaceutically acceptable salt thereof as described herein, and / or a checkpoint inhibitor as described herein in the method of the present invention. Thus, in certain embodiments, the product or kit includes instructions for using ketotifen or a pharmaceutically acceptable salt thereof as described herein, and / or a checkpoint inhibitor as described herein in a method for treating cancer (e.g., solid tumor) in a subject, comprising administering to the subject an effective amount of ketotifen or a pharmaceutically acceptable salt thereof as described herein, and / or a checkpoint inhibitor as described herein. In some embodiments of any of the aspects provided herein, the solid tumor is selected from the group consisting of mesothelioma, breast cancer, lung metastasis of breast cancer, sarcoma, pancreatic cancer, ovarian cancer, liver metastasis, prostate cancer, brain cancer, melanoma, renal cell carcinoma, colorectal cancer, hepatocellular carcinoma, lung cancer, head and neck squamous cell carcinoma, urothelial carcinoma, esophageal squamous cell carcinoma, gastric cancer, esophageal cancer, cervical cancer, Merkel cell carcinoma, endometrial carcinoma, and cutaneous squamous cell carcinoma. In some embodiments, the solid tumor is breast cancer. In some embodiments, the breast cancer has high tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the breast cancer is triple-negative breast cancer. In some embodiments, the solid tumor is lung metastasis of breast cancer. In some embodiments, the solid tumor is a sarcoma. In some embodiments, the solid tumor is an osteosarcoma. In some embodiments, the solid tumor is a fibrosarcoma. In some embodiments, the solid tumor is pancreatic cancer. In some embodiments, the solid tumor is an ovarian cancer. In some embodiments, the solid tumor is a liver metastasis. In some embodiments, the liver metastases are from colorectal cancer. In some embodiments, the solid tumor is prostate cancer.In some embodiments, the prostate cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the solid tumor is brain cancer. In some embodiments, the brain cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the solid tumor is melanoma. In some embodiments, the solid tumor is renal cell carcinoma. In some embodiments, the solid tumor is colorectal cancer. In some embodiments, the colorectal cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the colorectal cancer has low tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the colorectal cancer has high tumor endothelin-A receptor expression and low endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the solid tumor is hepatocellular carcinoma. In some embodiments, the solid tumor is lung cancer. In some embodiments, the lung cancer expresses endothelin-A receptor. In some embodiments, the lung cancer expresses endothelin-B receptor. In some embodiments, the lung cancer expresses both endothelin-A and endothelin-B receptors. In some embodiments, the lung cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the lung cancer has high tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the lung cancer has high tumor endothelin-A and endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the lung cancer is non-small cell lung cancer. In some embodiments, the lung cancer is small cell lung cancer. In some embodiments, the solid tumor is head and neck squamous cell carcinoma. In some embodiments, the solid tumor is urothelial carcinoma. In some embodiments, the solid tumor is esophageal squamous cell carcinoma. In some embodiments, the solid tumor is gastric cancer. In some embodiments, the solid tumor is esophageal cancer. In some embodiments, the solid tumor is cervical cancer. In some embodiments, the solid tumor is Merkel cell carcinoma. In some embodiments, the solid tumor is endometrial cancer. In some embodiments, the solid tumor is cutaneous squamous cell carcinoma. In some embodiments, the solid tumor is a cancer that has compressed blood vessels and / or is hypoperfused.In some embodiments, the solid tumor is a compressed vascular cancer. In some embodiments, the solid tumor is a hypoperfused cancer. In some embodiments, the compressed vascular and / or hypoperfused solid tumor is selected from the group consisting of breast cancer, lung metastasis of breast cancer, pancreatic cancer, ovarian cancer, and liver metastasis. In some embodiments, the compressed vascular and / or hypoperfused solid tumor is breast cancer. In some embodiments, the breast cancer has high tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the breast cancer is triple-negative breast cancer. In some embodiments, the compressed vascular and / or hypoperfused solid tumor is pancreatic cancer. In some embodiments, the compressed vascular and / or hypoperfused solid tumor is ovarian cancer. In some embodiments, the compressed vascular and / or hypoperfused solid tumor is a liver metastasis. In some embodiments, the liver metastasis is from colorectal cancer. In some embodiments, the compressed vascular and / or hypoperfused solid tumor is a lung metastasis. In some embodiments, the liver metastasis is from breast cancer. In some embodiments, the solid tumor is a cancer with endothelin receptor expression in tumor vasculature and / or fibroblasts. In some embodiments, the solid tumor is a cancer with endothelin receptor expression in tumor vasculature. In some embodiments, the solid tumor is a cancer with endothelin receptor expression in tumor fibroblasts. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is selected from the group consisting of pancreatic cancer, ovarian cancer, lung cancer, prostate cancer, brain cancer, breast cancer, and colorectal cancer. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is pancreatic cancer. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is ovarian cancer. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is lung cancer. In some embodiments, the lung cancer expresses endothelin-A receptor. In some embodiments, the lung cancer expresses endothelin-B receptor.In some embodiments, the lung cancer expresses both endothelin-A and endothelin-B receptors. In some embodiments, the lung cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the lung cancer has high tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the lung cancer has high tumor endothelin-A receptor and endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the lung cancer is non-small cell lung cancer. In some embodiments, the lung cancer is small cell lung cancer. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is prostate cancer. In some embodiments, the prostate cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is brain cancer. In some embodiments, the brain cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is breast cancer. In some embodiments, the breast cancer has high tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the breast cancer is triple-negative breast cancer. In some embodiments, the solid tumor is a lung metastasis from breast cancer. In some embodiments, the solid tumor with endothelin receptor expression in tumor vasculature and / or fibroblasts is colorectal cancer. In some embodiments, the colorectal cancer has high tumor endothelin-A receptor expression compared to non-tumor tissue. In some embodiments, the colorectal cancer has low tumor endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the colorectal cancer has high tumor endothelin-A receptor expression and low endothelin-B receptor expression compared to non-tumor tissue. In some embodiments, the subject is a human.

[0157] The product or kit may further comprise a container. Suitable containers include, for example, bottles, vials (e.g., two-chamber vials), syringes (e.g., one-chamber or two-chamber syringes), and test tubes. In some embodiments, the container is a vial. The container may be made of a variety of materials, such as glass or plastic. The container holds the formulation.

[0158] The product or kit may further include a label or package insert on or in association with the container, which may indicate instructions for reconstituting and / or using the formulation. The label or package insert may further indicate that the formulation is useful or intended for intraperitoneal injection, subcutaneous, intravenous (e.g., intravenous infusion), or other modes of administration for treating cancer (e.g., solid tumors) in a subject. The container holding the formulation may be a single-use vial or a multi-use vial that allows repeated administration of the reconstituted formulation. The product or kit may further include a second container that contains a suitable diluent. The product or kit may further include other materials that are desirable from a commercial, therapeutic, and user standpoint, including other buffers, diluents, bulking agents, needles, syringes, and package inserts with instructions for use.

[0159] The product or kit herein optionally further comprises a container containing a second medicament, and ketotifen or its pharmaceutically acceptable salt as described herein is the first medicament, and the product or kit further comprises instructions on a label or package insert for treating a subject with an effective amount of the second medicament.In some embodiments, the second medicament is a checkpoint inhibitor as described herein.In some embodiments, the label or package insert indicates that the first and second medicaments are administered sequentially or simultaneously as described herein.

[0160] In some embodiments, the ketotifen described herein and / or the chemotherapeutic agent described herein are present in the container as a lyophilized powder. In some embodiments, the ketotifen or its pharma- ceutically acceptable salt described herein and / or the checkpoint inhibitor described herein are present in the container as a lyophilized powder. In some embodiments, the lyophilized powder is a sealed container, such as a vial, an ampoule, or a sachet, indicating the amount of active agent. If the medicament is administered by injection, an ampoule of, for example, sterile water for injection or saline, can be provided as part of the kit, as needed, so that the components can be mixed before administration. Such kits can further include, if desired, one or more of various conventional pharmaceutical components, such as a container with one or more pharma- ceutically acceptable carriers, additional containers that will be readily apparent to those skilled in the art, and the like. Printed instructions, either as a package insert or label, indicating the amount of components to be administered, guidelines for administration, and / or guidelines for mixing the components, can also be included in the kit.

[0161] The present invention will be more fully understood with reference to the following examples. However, the examples should not be interpreted as limiting the scope of the present invention. The examples and embodiments described herein are for illustrative purposes only, and various modifications or changes in light thereof will be suggested to those skilled in the art, but they will be understood to be within the spirit and scope of this application and the appended claims. EXAMPLES

[0162] Example 1 Ketotifen inhibits angiogenesis in a dose-dependent manner and restores vascular perfusion within the tumor interstitium This example shows that ketotifen mediates angiogenic signaling and allows vascular normalization in two mouse models of fibrosarcoma and osteosarcoma.

[0163] Cell culture and animal tumor models Two mouse models of distinct sarcoma subtypes, fibrosarcoma (MCA205 cells) and osteosarcoma (K7M2wt cells), were used in this study.

[0164] MCA205 mouse fibrosarcoma cell line (SCC173, Millipore) was cultured in RPMI-1640 Growth Medium containing 2 mM L-glutamine, 1 mM sodium pyruvate, 10% fetal bovine serum, 1× non-essential amino acids (TMS-001-C, Sigma), 1% antibiotics (A5955, Sigma), and 1× β-mercaptoethanol. K7M2wt mouse osteosarcoma cell line (CRL2836™, ATCC®) was cultured in DMEM Growth Medium supplemented with 10% FBS and 1% antibiotics. All cells were maintained at 37° C. / 5% CO2.

[0165] Fibrosarcoma syngeneic tumor models were grown using 2.5 x 10 MCA205 cells in 50 µL of serum-free medium. 5 K7M2wt tumor pieces were subcutaneously implanted into the flanks of 6-week-old C57BL / 6 female mice. Osteosarcoma syngeneic tumor models were generated by implanting K7M2wt tumor pieces into the fat pads of 6-week-old BALB / c female mice. All mice were maintained specific pathogen-free and housed in a controlled temperature / humidity (22°C / 55%) environment with a 12-h light / dark cycle and had free access to food and water throughout the experimental period.

[0166] Ketotifen treatment and evaluation of tumor growth Each treatment group of 4-7 mice received sub-therapeutic doses of ketotifen (1, 5, 10 and 25 mg / kg) for 8 days. Ketotifen fumarate was dissolved in sterile saline (0.9% NaCl in ddH2O, w / v). Ketotifen was administered by intraperitoneal injection (ip) at 1 mg / kg, 5 mg / kg, 10 mg / kg, 25 mg / kg or an equivalent volume of diluent (control group) once daily for 8 days. Ketotifen treatment for MCA205 and K7M2wt tumor models was performed at 10 days after tumor growth, with mean tumor volumes of 60 mm2 and 60 mm3, respectively. 3 and 150mm 3 Starts when it reaches 600mm3 At study endpoint, primary tumors were surgically excised and stored in 1x PBS at -80°C until further processing. Tumor planar dimensions (x, y) were monitored every 2-3 days using digital calipers, and tumor volume was estimated from the volume of an ellipsoid, assuming the third dimension z was equal to sqrt(x, y).

[0167] After treatment, tumor growth (tumor volume and mass, see Figures 2A and 2C for MCA205 tumors and Figures 2B and 2D for K7M2wt tumors) was evaluated in mice from each sarcoma group. Subtherapeutic doses of ketotifen did not show antitumor effects in either mouse sarcoma model (Figures 2A-2D). Furthermore, subtherapeutic doses of ketotifen did not affect mast cell (MC) numbers, as determined by CD117 protein levels, which remained unaffected by ketotifen treatment (Figure 2E). Mouse body weights, thymus and spleen weights at the conclusion of the study indicated that the 10 mg / kg dose of ketotifen was tolerated (Figures 3A-3C for mice bearing MCA205 tumors; Figures 3D-3F for mice bearing K7M2wt tumors).

[0168] Hypoxia Research Mice bearing orthotopic MCA205 or K7M2wt sarcoma tumors were injected (intraperitoneally) with 60 mg / kg pimonidazole HCl 2 hours before tumor removal. Tumors were excised, washed twice for 10 min in 1× PBS, and incubated with 4% PFA overnight at 4°C. Fixative was aspirated and samples were washed twice for 10 min in 1× PBS. Tissue samples were dehydrated in successive ethanol steps and xylene before paraffin embedding. Serial sections (7 μm) of paraffin-embedded tissues were produced using a microtome (Accu-Cut SRM 200 Rotary Microtome, SAKURA), flattened in water, and dried overnight at 37°C. Sections were then deparaffinized and rehydrated as routinely processed for histology. Slides were microwaved in trisodium citrate, blocked in serum solution, and incubated with mouse anti-pimonidazole RED549 conjugated antibody (HP7-100Kit, 1:100) overnight at 4°C. The following day, slides were washed, nuclei were stained with DAPI, and tissue sections were mounted. Immunofluorescence images of slides were taken (Figure 1A), and the percentage of hypoxic areas across different treatment groups was normalized to DAPI staining.

[0169] The percentage of pimonidazole positives was significant across untreated, 1 mg / kg, 5 mg / kg, and 25 mg / kg ketotifen-treated mice and was significantly decreased in mice receiving the 10 mg / kg ketotifen dose (Figure 1A-1C). Intratumoral levels of hypoxia were determined by quantitating the fluorescent signal of pimonidazole adducts in images normalized to DAPI in both MCA205 (Figure 1B) and K7M2wt tumors (Figure 1C).

[0170] mRNA expression levels For this experiment, total RNA was isolated from breast tumors following a standard Trizol-based protocol (Invitrogen) and cDNA synthesis was performed using reverse transcriptase III (RT-III) enzyme and random hexamers (Invitrogen). Real-time polymerase chain reaction was performed using Sybr Fast Universal Master Mix (KAPA). Specific mouse primers used for gene expression analysis of IFN-γ, VEGF, and B-actin controls are listed in Table 1 below. Reactions were performed using a CFX-96 Real-time PCR Detection System (BioRad) under the following conditions: 2 min at 95°C, 2 s at 95°C, 20 s at 60°C, 1 s at 60°C, 39 cycles of steps 2-4. Real-time PCR analysis and calculation of changes in gene expression between compared groups were performed using ΔΔ Ct Methods were used. Relative gene expression was normalized based on the expression of β-actin. For qPCR analysis, 3–5 biological samples were used per treatment, with three technical replicates for each sample. [Table 1]

[0171] Ketotifen upregulated the mRNA levels of the antiangiogenic gene IFN-γ without affecting the mRNA levels of general angiogenic factors such as VEGF ( Fig. 1D ).

[0172] Interstitial fluid pressure Interstitial fluid pressure (IFP) was measured in vivo using the "wick-in-needle" technique after mice were anesthetized with ip injection of Avertin and prior to tumor resection. Additional information regarding the wick-in-needle technique can be found in Dong et al., Involvement of mast cell chymase in burn wound healing in hamsters 2013;5:643-7 and Shankaran et al. IFNγ and lymphocytes prevent primary tumour development and shape tumour immunogenicity 2001;410:1107-11, the contents of which are incorporated herein by reference in their entirety.

[0173] All doses of ketotifen reduced IFP, with the 10 mg / kg dose showing the greatest effect ( Figure 1 E), further highlighting its ability to prevent fluid leakage into the tumor microenvironment (TME) by re-establishing vascular function.

[0174] Overall, these results indicate that ketotifen inhibits angiogenesis and restores vascular perfusion in the tumor interstitium in a dose-dependent manner, with a ketotifen dose of 10 mg / kg producing a robust effect.

[0175] Example 2 Ketotifen reduces intratumoral stiffness by inhibiting extracellular matrix formation This example shows that ketotifen induces stress reduction within the TME in a mouse sarcoma model, thereby facilitating vascular decompression, a measure of intratumoral stiffness.

[0176] Ultrasound elastography measurements Cell cultures and animal sarcoma tumor models were prepared as described above, and an overview of the study is shown in Figure 4A.

[0177] The mechano-therapeutic ability of ketotifen in reducing stromal stiffness was primarily evaluated non-invasively and longitudinally using ultrasound elastography. Briefly, evaluation of tumor modulus, which was elastic by shear wave elastography using a Philips EPIQ Elite Ultrasound scanner containing an eL18-4 linear array, approved for clinical scanning, was performed as described in a previous study. Modulus values ​​were obtained for the area of ​​the region of interest (ROI) showing the highest shear wave quality. Modulus values ​​presented are averages within the tumor area. Shear wave imaging of MCA205 tumors was performed before ketotifen treatment on day 7 to determine baseline levels of tumor stiffness 3 days after treatment (day 11) and on the last day of treatment (day 15) just before tumor removal. Ultrasound imaging of K7M2wt tumors was performed on days 24, 28, and 31. The effect of ketotifen on the chemo-immunotherapy combination on MCA205 tissue elasticity was assessed 7 days before the first cycle of doxorubicin + anti-PD-L1 treatment, 1 day after the second cycle (day 11), and 2 days after the last cycle (day 15). Ultrasound imaging of K7M2wt tumors was performed on day 22 (i.e., 4 days after daily ketotifen and before the first cycle of combination treatment) and at the completion of treatment (day 32).

[0178] Ketotifen at a dose of 10 mg / kg most reduced tissue stiffness in mice bearing MCA205 fibrosarcoma tumors, with Young's modulus values ​​reaching 20 kPa, similar to the elasticity of healthy tissue (Figure 4B). K7M2wt osteosarcoma tumors similarly benefited from 10 mg / kg daily ketotifen (Figure 4C). Microscopic elasticity measurements obtained by AFM were consistent with the macroscopic data.

[0179] Vascular and functional perfusion Vascular and functional perfusion were measured simultaneously using contrast-enhanced ultrasound on days 3 and 7 during the course of ketotifen treatment in mice bearing MCA205 and K7M2wt tumors. Tumor perfusion was assessed after a bolus injection of 8 μl of SonoVue / Lumason contrast agent (Bracco Diagnostics, Geneva, Switzerland). SonoVue contains sulfur hexafluoride microbubbles encapsulated in a phospholipid shell and with a mean diameter of 2.5 μm. Because tail vein microbubble injections tend to have high variability, it is delivered as a retro-orbital injection. Before each ultrasound application, mice were anesthetized by i.p. injection of Avertin (200 mg / kg). As shown in Figures 4D-4G, ketotifen at a dose of 10 mg / kg caused a significant increase in vascular and functional perfusion in both sarcoma subtypes.

[0180] Collagen staining Because tissue stiffness is driven in part by the degree of collagen expression and cross-linking by cancer-associated fibroblasts (CAFs), intratumoral collagen levels were assessed by histological staining using picrosirius red.

[0181] Briefly, tumors were excised from four mice, washed twice for 10 min in 1×PBS, and incubated overnight at 4° C. with 4% PFA. Fixative was aspirated, and samples were washed twice for 10 min in 1×PBS. Tissue samples were dehydrated in successive ethanol steps and xylene, followed by paraffin embedding. Serial sections (7 μm) of paraffin-embedded tissues were produced using a microtome (Accu-Cut SRM 200 Rotary Microtome, SAKURA), flattened in water, and dried overnight at 37° C. Sections were then deparaffinized and rehydrated as routinely processed for histology. After deparaffinization and rehydration, tissue sections were immersed in picrosirius red staining solution for 1 h at room temperature. Tissue sections were then rinsed with two changes of acetic acid, followed by two changes of absolute ethanol, and finally mounted with DPX mounting medium (Sigma) for histology. The collagen fibers stained red, while the rest of the tissue was pale yellow.

[0182] Ketotifen at a dose of 10 mg / kg caused a significant decrease in collagen deposition in both sarcoma subtypes (Figures 5A and 5C for MCA205 tumors; Figures 5B and 5D for K7M2wt tumors).

[0183] Expansion of CAFs CAF proliferation was assessed by immunostaining for the presence of alpha smooth muscle actin (α-SMA) (ab5694, Abcam; 1:50 dilution) protein and Ki-67 (Abcam; 1:200 dilution) proliferation markers after antigen retrieval, incubation in serum solution (10% FBS, 3% donkey serum in 1×PBS) for 1 hour, and incubation in blocking solution for endogenous mouse IgG (Abcam; 1:100 dilution) for 1 hour. After washing with TBS-T (0.25% Tween®-20), Alexa Fluor-647 anti-rabbit IgG (H+L) (Invitrogen; 1:400 dilution) and Fluor-488 anti-rat IgG (H+L) (Invitrogen; 1:400 dilution) secondary antibodies and DAPR (Sigma, 1:100 dilution of 1 mg / mL stock) were applied for 1 hour at room temperature. Sections were then mounted on microscope slides using ProLong gold antifade mounting medium (Invitrogen) and covered with a coverslip. The percentage of CAF proliferation was determined as the ratio of the total CAF signal (α-SMA) to the total CAF signal (α-SMA) of overlapping α-SMA (green) and Ki67 (red) positive signals. + ) and CAF was expressed as a ratio.

[0184] We verified the extracellular matrix (ECM) alterations indicated by reduced collagen deposition after 10 mg / kg ketotifen treatment (Figures 5A-5D) and assessed CAF levels and proliferation activity in Figures 5E-5I.

[0185] Figures 5H-5I and 6C-6D show quantification of the percentage of area positive for α-SMA staining (Figures 5H and 6C) and Ki-67 staining (Figures 5I and 6D) in mice bearing MCA205 tumors and mice bearing K7M2wt tumors in the representative immunofluorescence images of Figures 5E and 6A, respectively. As shown by immunofluorescence staining for α-SMA and Ki-67, respectively, in fibrosarcoma tumors (Figures 5E-5F and 6A-6B), 10 mg / kg ketotifen treatment results in a decrease in CAF levels and proliferative activity in both mouse sarcoma models.

[0186] mRNA expression levels mRNA expression levels were assessed in both mouse sarcoma models by qPCR as described above. The specific mouse primers used for gene expression analysis are listed in Table 2. [Table 2]

[0187] Ketotifen reduced the mRNA levels of genes encoding collagen I (Col1A1) and connective tissue growth factor (CTGR), but not to a statistically significant extent, and had no effect on hyaluronan synthase 2 (Has2) and hyaluronan synthase 3 (Has3) in MCA205 tumors (Figure 5G and Figure 7A). A similar effect was observed in K7M2wt tumors (Figure 7B).

[0188] Hyaluronan Evaluation Besides collagen, other structural components of the tumor stroma can exert pressure and distort blood vessels. Such an example is hyaluronan (e.g., hyaluronic acid), which binds and retains gel-like regions that generate excess fluid within the tissue. Therefore, hyaluronic acid staining was performed in both sarcoma mouse models.

[0189] Tumors were excised from mice, washed twice for 10 min in 1×PBS, and incubated overnight at 4° C. with 4% PFA. Fixative was aspirated and samples were washed twice for 10 min in 1×PBS. Tissue samples were dehydrated in successive ethanol steps and xylene before being embedded in paraffin. Serial sections (7 μm) of paraffin-embedded tissues were produced using a microtome (Accu-Cut SRM 200 Rotary Microtome, SAKURA), flattened in water, and dried overnight at 37° C. Sections were then deparaffinized and rehydrated as routinely processed for histology. Tissue sections were subjected to antigen retrieval (microwave heat treatment with trisodium citrate, pH 6, 20 min), washed with 1×TBS / 0.025% Triton®X-100 (TBS-T), and incubated in blocking solution (2% BSA, 0.2% Triton®100x) at room temperature for 2 h. Slides were then incubated with biotinylated hyaluronan binding protein (b-HABP) (amsbio, 1:100) overnight at 4° C. After incubation with streptavidin-FITC conjugate (Invitrogen, 1:1000), hyaluronan was detected and nuclei were stained with DAPI stain for 1 h at room temperature in the dark. The percentage of areas positive for hyaluronan across different treatments was normalized to DAPI staining.

[0190] Immunofluorescence staining of hyaluronan binding protein 1 (HABP1) showed that daily administration of ketotifen at a dose of 10 mg / kg reduced the expression of hyaluronan protein (Figures 5J-5K and 6E-F).

[0191] Tumor opening Tumor opening experiments were performed using the post-section opening value as an indicator of the amount of residual stress contained within the tissue. MCA205 tumors treated with moderate doses of ketotifen (5 mg / kg and 10 mg / kg) showed smaller openings, suggesting a reduction in solid stress (Figure 5L).

[0192] Overall, these results indicate that ketotifen (10 mg / kg) reduces intratumoral stiffness in sarcoma tumors and that daily ketotifen administration induces ECM remodeling by inhibiting collagen and hyaluronan formation.

[0193] Example 3 Ketotifen pretreatment enhances the antitumor efficacy of immunotherapy by promoting T cell recruitment and cytotoxic immune responses This example shows that ketotifen enhances anti-tumor immune responses in combination treatment. In particular, this example shows that a 10 mg / kg daily regimen of ketotifen enhances the anti-tumor immune responses of doxorubicin / anti-PD-L1 combination treatment.

[0194] Animal tumor models Mice bearing sarcoma tumors were pretreated with 10 mg / kg daily ketotifen followed by 3 or 4 doses of neoadjuvant chemotherapy in combination with an immune checkpoint inhibitor (ICI) anti-PD-L1 antibody (Figure 8A).

[0195] Fibrosarcoma syngeneic tumor models were generated and ketotifen was prepared as described above. Mouse monoclonal anti-PD-L1 (B7-H1, clone 10F.9G2, BioXCell) and rat IgG2b isotype control, anti-keyhole limpet hemocyanin (LTF-2, BioXCell) were dissolved in the recommended InVivoPure pH 7.0 dilution buffer (IP0070, BioXCell). Doxorubicin hydrochloride was obtained from Nicosia General Hospital as a ready-made solution at 2 mg / ml. Anti-PD-L1 was administered at a final dose of 10 mg / kg and doxorubicin was administered at 5 mg / kg.

[0196] Mice bearing MCA205 tumors were cultured at 100-mL sham-treated 10-well plates with an average tumor size of 40 mm. 3 After tumors reached 150 mm , patients were pretreated daily with 10 mg / kg ketotifen or an equal volume of diluent (control group) before neoadjuvant chemotherapy treatment. 3Doxorubicin and anti-PD-L1 combination treatment was administered as an ip injection every 3 days (days 7, 10, and 13) for three doses, beginning when the average size of tumors reached 10 mm Hg (day 7). Daily administration of ketotifen was continued until the end of doxorubicin-anti-PD-L1 combination treatment.

[0197] Primary tumor was 700 mm on day 16. 3 Once the primary tumors reached an average tumor size of 70 or 80 mm, the primary tumors were excised and preserved, and the mice were monitored for rechallenge experiments. Similarly, K7M2wt tumors were excised and preserved until the average tumor size reached 70 or 80 mm. 3 When tumors reached 150 mm (day 18), they were pretreated with 10 mg / kg ketotifen or an equal volume of diluent (control group) daily until the end of preoperative chemotherapy treatment. 3 Doxorubicin and anti-PD-L1 combination treatment was initiated when tumors reached an average size of 550 mm (day 22) and repeated on days 25, 28, and 31. 3 The study was terminated when the tumor reached a mean volume of 0.01 mg / kg / day (day 33). Mice were sacrificed and tumors were collected for ex vivo analysis.

[0198] Tumor characterization Neither anti-PD-L1 nor doxorubicin monotherapy affected tumor growth in fibrosarcoma MCA205 tumors, but their combination with ketotifen induced a significant anti-tumor response (Figure 8B and Figure 8F). In an osteosarcoma tumor model (K7M2wt), a similar decrease in relative tumor growth was observed in mice receiving the doxorubicin-anti-PD-L1 combination, as well as in all combinations of ketotifen mechanical therapy with the triple combination of ketotifen-doxorubicin and anti-PD-L1, which caused the greatest attenuation of proliferation (Figure 8C).

[0199] To support the hypothesis that the increased efficacy of cytotoxic treatment was due to a ketotifen effect, tissue stiffness was measured before the first cycle of chemotherapy or immunotherapy in the ketotifen and non-ketotifen treated groups, and these values ​​were correlated with relative tumor volume, defined as the ratio of the final volume to the volume obtained before the first cycle of chemotherapy-immunotherapy treatment. Ketotifen-induced reduction in tissue stiffness was evaluated for correlation with tumor response to neoadjuvant. Young's modulus measurements were determined as previously described and recorded 4 days after ketotifen administration. Young's modulus measurements positively correlate with relative tumor volume across doxorubicin and anti-PD-L1 monotherapy and combination treatment groups in both MCA205 (Figure 8D) and K7M2wt tumors (Figure 8E). Response to therapy is improved when stiffness values ​​fall below 30 kPa for both tumor models. These results show a linear negative correlation between the elastic modulus measured at the start of the cytotoxic treatment and antitumor efficacy as indicated by the R2 value of the best linear fit (Figure 8D and Figure 8E).

[0200] Re-challenge experiment Because doxorubicin has the ability to establish a favorable immunogenic state within the TME that may affect the efficacy of immunotherapy, we investigated whether this immune response would be preserved upon rechallenge of long-term survivors with the same cell line. Mice were considered long-term survivors if no tumors were detectable 80 days after surgical resection of the MCA205 primary tumor. Five long-term survivors from each treatment group were administered 2.5 × 10 5 Mice were further rechallenged by subcutaneous injection of 100 MCA205 cells in the flank area. Cells were prepared in 50 μl of PBS. Naive mice were implanted simultaneously as controls.

[0201] As shown in Figure 8G, interestingly, no tumor growth occurred in the ketotifen-doxorubicin-anti-PD-L1 group, indicating that a memory response upon tumor antigen recognition, i.e., the development of an adaptive memory response, is induced upon tumor antigen encounter. Conversely, all naive mice treated with ketotifen alone developed progressively growing tumors. Similarly, large tumors were formed in four mice in the doxorubicin and anti-PD-L1 group and in three mice in the doxorubicin-anti-PD-L1 group. In the ketotifen-doxorubicin and ketotifen-anti-PD-L1 groups, only one of five mice developed fibrosarcoma. Similar to the efficacy hierarchy observed in tumor growth measurements, the ketotifen-doxorubicin-anti-PD-L1 combination showed therapeutic superiority over doxorubicin-anti-PD-L1 without ketotifen, resulting in sustained remission.

[0202] Taken together, these findings indicate that pretreatment with ketotifen is a prerequisite for establishing a favorable immunogenic state within the TME that allows for potent antitumor effects leading to immune memory and allows for doxorubicin and anti-PD-L1 antibody treatment to confer immune memory.

[0203] Example 4 Ketotifen attenuates hypoxia and restores intratumoral T cell infiltration by upregulating immune cell adhesion to blood vessels This example demonstrates the contribution of ketotifen and ketotifen combination treatment in stimulating the immune response.

[0204] Flow cytometry On day 16 of the doxorubicin-anti-PD-L1 combination study (e.g., the study outlined in Figure 8A), MCA205 tumors were harvested in 1x HBSS (Biosera), minced into fine fragments, and incubated with 1mg / mL collagenase D and 0.5mg / ml DNaseI (Roche) for 30 minutes in a 37°C shaker. Enzyme digestion was stopped by the addition of RPMI medium containing 10% FBS and 1% antibiotic / antimycotic solution. Tissue homogenates were then filtered through a 40μm cell strainer. Red blood cells were removed with ACK lysis buffer (A1049201, Gibco) and single cell suspensions were collected and 1x10 per sample. 6 A final concentration of 1:1000 cells was used. Cells were then incubated with a fixable viability dye (eBioscience, 1:1000) for 10 min on ice for live cell gating. Non-specific antibody binding was blocked after incubation with rat anti-mouse CD16 / CD32 mAb (BD Bioscience) for 20 min on ice.

[0205] The anti-mouse antibodies used in the experiments were CD45-V500 (BD Bioscience), CD3-PE (BD Bioscience), CD4-PerCP Cy5.5 (Invitrogen), CD25-PE-Cy7 (BD Bioscience), CD127-APC (BioLegend) and CD8a-e450 (eBioscience).

[0206] Foxp3-AF488 (BD Bioscience) staining was performed after fixation and permeabilization process using Foxp3 / transcription factor staining buffer set (00-5523-00, eBioscience). For MDSC staining, Gr-1-PE (BioLegend) and CD11b-e450 (eBioscience) were used. Flow cytometry data was acquired using a BD FACSLyric flow cytometer and analyzed using BD FACSuite™ software. Data presented are representative of singlet live cells. Figures 11A-11B and 12 provide details regarding the gating strategy for the flow cytometry experiments provided herein.

[0207] The total T cell population was divided into 10 subpopulations, primarily CD3 + After defining by expression of the pan-T cell marker protein, CD8 + Cytotoxic or regulatory T cell subtypes (CD3 + CD4 + CD25 hi CD127 lo Foxp3 + ) (Figure 9A). There was a statistically significant increase in total T cell recruitment in the doxorubicin-anti-PD-L1 and ketotifen-doxorubicin-anti-PD-L1 treated groups. Although the magnitude of the increase in the CD8 / Treg ratio was small compared to more immunogenic cancers such as breast cancer, it was associated with a response to ICI. However, in K7M2wt tumors, only the group of mice receiving ketotifen prior to the neoadjuvant regimen showed an increase in T cell infiltration, as shown by immunofluorescence staining for CD3 protein (Figure 10D). CD8 + The ratio of T cells to immunosuppressive Tregs was increased after combining doxorubicin with ketotifen or / and immunotherapy treatment (FIG. 9B).

[0208] Immunofluorescence and mRNA expression Immunofluorescence staining for Ki67 and CD8, performed as described above, revealed recruited CD8 in osteosarcoma tumors (K7M2wt tumors).+ The activation state of T cells was confirmed (Figures 9C-9D). K7M2wt tissue sections were then stained for pimonidazole adducts as a measure of hypoxia formation. Only the group receiving ketotife...

Claims

1. A pharmaceutical composition for use in a method for treating a solid tumor in a subject in need thereof, comprising ketotifen or a pharmaceutically acceptable salt thereof, wherein the treatment comprises administering to the subject the pharmaceutical composition in combination with a checkpoint inhibitor.

2. A pharmaceutical composition for use in a method for treating a solid tumor in a subject in need thereof, comprising a checkpoint inhibitor, wherein the treatment comprises administering to the subject the pharmaceutical composition in combination with ketotifen or a pharmaceutically acceptable salt thereof.

3. A pharmaceutical composition for use in a method for treating a solid tumor in a subject in need thereof, comprising a combination of ketotifen or a pharmaceutically acceptable salt thereof and a checkpoint inhibitor.

4. 1. A pharmaceutical composition for use in a method for increasing blood flow to a solid tumor in a subject, the method comprising administering to the subject the pharmaceutical composition in combination with a checkpoint inhibitor, the pharmaceutical composition comprising ketotifen or a pharmaceutically acceptable salt thereof.

5. 5. The pharmaceutical composition of claim 4, wherein blood flow is measured using ultrasound-based blood flow measurements or using histology techniques to measure hypoxia.

6. A pharmaceutical composition described in any one of claims 1 to 4, wherein administering the pharmaceutical composition increases the number of anti-tumor T cells co-localized with the solid tumor.

7. A pharmaceutical composition described in any one of claims 1 to 4, wherein administering the pharmaceutical composition reduces the tissue hardness of the solid tumor.

8. 8. The pharmaceutical composition of claim 7, wherein the tissue stiffness of the solid tumor is measured using ultrasound elastography.

9. A pharmaceutical composition described in any one of claims 1 to 4, wherein administering the pharmaceutical composition reduces the level of extracellular matrix proteins in the solid tumor.

10. 10. The pharmaceutical composition of claim 9, wherein the extracellular matrix protein is collagen I or hyaluronan binding protein (HABP).

11. A pharmaceutical composition described in any one of claims 1 to 4, wherein administering the pharmaceutical composition reduces hypoxia in the solid tumor.

12. 5. The pharmaceutical composition of any one of claims 1 to 4, wherein the checkpoint inhibitor inhibits a checkpoint protein selected from the group consisting of CTLA-4, PD-1, PD-L1, PD-L2, B7-H3, B7-H4, BMA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, and B-7 family ligand, or a combination thereof.

13. 13. The pharmaceutical composition of claim 12, wherein the checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA4 antibody, or a combination thereof.

14. 13. The pharmaceutical composition of claim 12, wherein the checkpoint inhibitor is selected from the group consisting of MEDI0680, AMP-224, nivolumab, pembrolizumab, pidilizumab, MEDI4736, atezolizumab, ipilimumab, tremelimumab, and BMS-936559.

15. The pharmaceutical composition according to any one of claims 1 to 4, wherein the ketotifen or a pharmaceutically acceptable salt thereof is administered to the subject once a day.

16. The pharmaceutical composition according to any one of claims 1 to 4, wherein the ketotifen or a pharmaceutically acceptable salt thereof is administered to the subject at a dose of about 0.01 mg / kg to about 5 mg / kg.

17. The pharmaceutical composition according to any one of claims 1 to 4, wherein the ketotifen or a pharmaceutically acceptable salt thereof is administered to the subject at a dose of about 100 mg to about 1200 mg.

18. 5. The pharmaceutical composition of any one of claims 1, 2 and 4, wherein the ketotifen or a pharmaceutically acceptable salt thereof is administered to the subject prior to administering the checkpoint inhibitor to the subject.

19. 5. The pharmaceutical composition of any one of claims 1, 2 and 4, wherein the administration of ketotifen or a pharmaceutically acceptable salt thereof to the subject is maintained for at least a portion of the period during which the checkpoint inhibitor is administered to the subject.

20. The pharmaceutical composition of any one of claims 1 to 4, wherein one or more therapeutic effects in the subject are improved compared to baseline after administration of the ketotifen or a pharmaceutically acceptable salt thereof and the checkpoint inhibitor, and the one or more therapeutic effects are selected from the group consisting of size of a tumor derived from cancer, objective response rate, duration of response, time to response, progression-free survival, and overall survival.

21. The pharmaceutical composition described in any one of claims 1 to 4, wherein the size of a tumor derived from cancer is reduced by at least about 10% compared to the size of the tumor derived from the cancer before administration of the ketotifen or its pharmaceutically acceptable salt and the checkpoint inhibitor.

22. After administration of ketotifen or a pharmaceutically acceptable salt thereof and the checkpoint inhibitor, the objective response rate is at least about 20%; the subject exhibits a progression-free survival of at least about 1 month; or The pharmaceutical composition of any one of claims 1 to 4, wherein the subject exhibits an overall survival of at least about 1 month.

23. 5. The pharmaceutical composition of claim 1, wherein the duration of response to the ketotifen or a pharmaceutically acceptable salt thereof and the checkpoint inhibitor is at least about 1 month after administration of the ketotifen or a pharmaceutically acceptable salt thereof and the checkpoint inhibitor.

24. 5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the solid tumor is selected from the group consisting of mesothelioma, breast cancer, lung metastasis of breast cancer, sarcoma, pancreatic cancer, ovarian cancer, liver metastasis, prostate cancer, brain cancer, melanoma, renal cell carcinoma, colorectal cancer, hepatocellular carcinoma, lung cancer, head and neck squamous cell carcinoma, urothelial carcinoma, esophageal squamous cell carcinoma, gastric cancer, esophageal cancer, cervical cancer, Merkel cell carcinoma, endometrial cancer, and cutaneous squamous cell carcinoma.

25. 25. The pharmaceutical composition of claim 24, wherein the solid tumor is a sarcoma.

26. 26. The pharmaceutical composition of claim 25, wherein the sarcoma is osteosarcoma or fibrosarcoma.

27. The pharmaceutical composition according to any one of claims 1 to 4, wherein the subject is a human.

28. The pharmaceutical composition of any one of claims 1 to 4, wherein the method further comprises administering an additional chemotherapeutic agent.

29. 29. The pharmaceutical composition of claim 28, wherein the additional chemotherapeutic agent is doxorubicin or an analog or derivative thereof.

30. (a) an effective amount of ketotifen or a pharmaceutically acceptable salt thereof; (b) an effective amount of a checkpoint inhibitor; and (c) Instructions for using the ketotifen or a pharmaceutically acceptable salt thereof and the checkpoint inhibitor in the method of any one of claims 1 to 4. Kit including:

31. The pharmaceutical composition of any one of claims 1 to 4, wherein the subject is selected for the method based on an increase in blood flow and / or a decrease in hardness of the solid tumor in response to ketotifen or a pharmaceutically acceptable salt thereof.

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