Oncology combination therapy and methods of use

Abstract

Disclosed herein is the treatment of cancer by administering plinabulin in combination with an immune checkpoint inhibitor.

Description

ONCOLOGY COMBINATION THERAPY AND METHODS OF USECROSS REFERENCE TO RELATED APPLIATIONS

[0001] This patent application claims the benefit of priority to U.S. Provisional Application No. 63 / 523,609, filed June 27, 2023, U.S. Provisional Application No. 63 / 589,579, filed October 11, 2023, and U.S. Provisional Application No. 63 / 631,251, filed April 8, 2024. All of the foregoing applications are fully incorporated herein by reference in their entireties for all purposes.BACKGROUNDField

[0002] The present disclosure relates to the field of chemistry and medicine. More specifically, it relates to combination therapies comprising plinabulin, an immune checkpoint inhibitor and radiation therapy for the treatment of cancer.Description of the Related Art

[0003] Human cancers harbor numerous genetic and epigenetic alterations, generating neoantigens potentially recognizable by the immune system (Sjoblom et al, 2006). The adaptive immune system, comprised of T and B lymphocytes, has powerful anti-cancer potential, with a broad capacity and exquisite specificity to respond to diverse tumor antigens.

[0004] Recent cancer immunotherapy research has focused substantial effort on approaches that enhance anti-tumor immunity by mediating an adaptive immune response to relevant antigens, providing specific immune- stimulatory agents such as immune checkpoint inhibitors. While cancer remains as an incurable disease for the great majority of patients, there exists a particular need for developing effective therapeutic agents and treatment regimens that can be used in cancer therapy. Patients receiving immune checkpoint inhibitors may initially respond to these checkpoint inhibitors, but at a later time become unresponsive (resistant) to these checkpoint inhibitors, or may not respond to these checkpoint inhibitors from the start of their administration (non-responders).SUMMARY

[0005] In a first aspect of the present disclosure, provided herein is a method of treating cancer in a subject, the method comprising a treatment cycle comprising the steps of:(i) administering one or more immune checkpoint inhibitor to the subject;(ii) administering plinabulin or a pharmaceutically acceptable salt thereof to the subject;(iii) obtaining a biological sample from the subject; and(iv) determining the levels of one or more biomarkers or levels of one or more cells in the biological sample.

[0006] In some embodiments of the first aspect, treatment with the one or more immune checkpoint inhibitor and plinabulin may be discontinued if the level of the one or more biomarker or one or more cells in the biological sample is above or below a predetermined threshold level, or if a score determined by mathematically combining levels of two or more biomarkers or two or more cells is above or below a predetermined threshold value.

[0007] In some embodiments of the first aspect, the one or more biomarker is CCR7, CD40, CD80, CD83, CD86, GEF-H1, IL-2, IFNy, IL-6, IL-12p70, IL-12p40, IL-13, IL-17A, IL-23, G-CSF, PD-L1, IL-8 and IFN-0, or a combination thereof. In some embodiments, the one or more biomarker includes CCR7. In some embodiments, the one or more biomarker includes CD40. In some embodiments, the one or more biomarker includes CD80. In some embodiments, the one or more biomarker includes CD83. In some embodiments, the one or more biomarker includes CD86. In some embodiments, the one or more biomarker includes GEF-H1.

[0008] In some embodiments of the first aspect, the one or more immune checkpoint inhibitor is pembrolizumab, nivolumab, cemiplimab, atezolizumab, avelumab, pidilizumab, ipilimumab, BMS 936559, durvalumab, camrelizumab, dostarlimab, tislelizumab, sintilimab, toripalimab, or a combination thereof. In some embodiments, the one or more immune checkpoint inhibitor is avelumab. In other embodiments, the one or more immune checkpoint inhibitor is atezolizumab. In yet other embodiments, the one or more immune checkpoint inhibitor is durvalumab. In still yet other embodiments, the one or more immune checkpoint inhibitor is nivolumab. In some embodiments, the one or more immune checkpoint inhibitor is pembrolizumab. In some embodiments, the one or more immune checkpoint inhibitor is camrelizumab. In some embodiments, the one or more immunecheckpoint inhibitor is dostarlimab. In some embodiments, the one or more immune checkpoint inhibitor is tislclizumab. In some embodiments, the one or more immune checkpoint inhibitor is sintilimab. In some embodiments, the one or more immune checkpoint inhibitor is toripalimab.

[0009] In some embodiments of the first aspect, wherein the treatment cycle is 21 days. In other embodiments, the treatment cycle is 28 days.

[0010] In some embodiments of the first aspect, the one or more immune checkpoint inhibitor may be administered intravenously.

[0011] In some embodiments of the first aspect, the one or more immune checkpoint inhibitor is administered on day 1 of the treatment cycle. In other embodiments, the one or more immune checkpoint inhibitor is administered on day 15 of the treatment cycle. In some embodiments, the one or more immune checkpoint inhibitor to the subject in an amount of from 50 mg to 2000 mg

[0012] In some embodiments of the first aspect, the one or more immune checkpoint inhibitor is administered over a period of about 10 minutes to about 180 minutes. In some embodiments, the one or more immune checkpoint inhibitor is administered over a period of about 10 minutes to about 120 minutes. In other embodiments, the one or more immune checkpoint inhibitor is administered over a period of about 30 minutes to about 90 minutes. In some embodiments, the one or more immune checkpoint inhibitor is administered over a period of about 30 minutes. In other embodiments, the one or more immune checkpoint inhibitor is administered over a period of about 60 minutes.

[0013] In some embodiments of the first aspect, the plinabulin is administered on day 1 of the treatment cycle. In some embodiments, the plinabulin is administered on day 4 of the treatment cycle. In some embodiments, the plinabulin is administered on day 1 and day 4 of the treatment cycle.

[0014] In some embodiments of the first aspect, the dose of plinabulin is from about 10 mg / m2to about 50 mg / m2. In some embodiments, the dose of plinabulin is about 20 mg / m2. In other embodiments, the dose of plinabulin is about 30 mg / m2.

[0015] In some embodiments of the first aspect, the plinabulin is administered over a period of 10 minutes to 128 minutes. In some embodiments, the plinabulin is administered over a period of about 10 minutes to about 120 minutes. In other embodiments, the plinabulinis administered over a period of 20 minutes to 90 minutes. In some embodiments, the plinabulin is administered over a period of 30 minutes to 60 minutes.

[0016] In some embodiments of the first aspect, when the one or more immune checkpoint inhibitor and the plinabulin are administered on the same day, the plinabulin is administered from about 0.5 hours to about 3 hours after completion of administration of the one or more immune checkpoint inhibitor. In some embodiments, the plinabulin is administered from about 1 hour to about 2 hours after completion of administration of the one or more immune checkpoint inhibitor.

[0017] In some embodiments of the first aspect, the method further comprises administering radiation therapy to the subject. In some embodiments, radiation therapy is administered in one to ten fractions. In other embodiments, radiation therapy is administered three to five fractions. In some embodiments, radiation therapy is administered in three fractions. In other embodiments, radiation therapy is administered in four fractions. In yet other embodiments, radiation therapy is administered in five fractions. In some embodiments, the total dose of radiation administered is from about 1 Gy to about 20 Gy. In other embodiments, the total dose of radiation administered is from about 2 Gy to about 15 Gy. In yet other embodiments, the total dose of radiation administered is from about 4 Gy to about 15 Gy. In some embodiments, the total dose of radiation administered is about 4 Gy. In some embodiments, the total dose of radiation administered is about 8 Gy. In some embodiments, the total dose of radiation administered is about 12.5 Gy.

[0018] In some embodiments of the first aspect, radiation therapy is administered on days 1, 2, and 3 of the treatment cycle. In other embodiments, radiation therapy is administered on days 1, 2, 3, and 4 of the treatment cycle. In yet other embodiments, radiation therapy is administered on days 1, 2, 3, 4, and 5 of the treatment cycle. In some embodiments, when radiation therapy and plinabulin are administered on the same day, the plinabulin is administered from about 3 hours to about 12 hours after completion of administration of the radiation therapy.

[0019] In some embodiments of the first aspect, the biological sample is a blood sample. In some such embodiments, the blood sample is a peripheral blood sample. In other embodiments, the biological sample is a tumor biopsy.

[0020] In some embodiments of the first aspect, the one or more cells are dendritic cells (DC) and their subsets (c.g., myeloid dendritic cells (mDC), plasmacytoid dendritic cells (pDC) conventional dendritic cells (eDC)); or wherein the one or more cells are selected from the group consisting of CD3+ T-cells, CD4+ T-cells CD8+ T-cells, B-cells, IgA-i- plasma cells, IgG-i- plasma cells, and IgM+ plasma cells; or wherein the one or more cells are selected from the group consisting of CD16+ monocytes, CD14+ monocytes, natural killer (NK) cells, granzyme K+ T cells, CD8+ TRM cells, CD8+ TEM cells, CD4+ TCM cells, CD4+ TEM cells, CD4+ naive T-cells, naive B-cells, cDCl cells cDC2 cells, and DC3 cells. In some embodiments, the one or more cells are monocyte-derived macrophage cells (MoMac), and interstitial macrophages (IM). In other embodiments, the one or more cells are immune cells from peripheral blood mononuclear cells (PBMCs).

[0021] In some embodiments of the first aspect, the method comprises determining the level of one or more dendritic cells and their subsets in the biological sample expressing one or more biomarkers. In some embodiments, the dendritic cells are plasmacytoid dendritic cells (pDC), conventional dendritic cells (eDC), or myeloid dendritic cells (mDC). In some embodiments, the one or more biomarker is CCR7, CD40, CD80, CD83, CD86, GEF-H1, IL- 2, IFNy, IL-6, IL-12p70, IL-12p40, IL-13, IL-17A, IL-23, G-CSF, IL-8, IFN-0, or a combination thereof. In some embodiments, the one or more biomarker is CCR7, CD40, CD80, CD83, CD86, or a combination thereof. In other embodiments, one or more biomarker is GEF-H1 immune-activation gene signature score (i.e., GEF-H1 immune activation score).

[0022] In some embodiments of the first aspect, the biological sample obtained on day 4 of the treatment cycle.

[0023] In some embodiments of the first aspect, the cancer is selected from a breast cancer, a bladder cancer, a glioma, a glioblastoma, a head and neck cancer, a non-small cell lung cancer, a small cell lung cancer, recurrent small cell lung cancer (SCLC), a colorectal cancer, a gastrointestinal stromal tumor, a gastroesophageal carcinoma, a renal cell cancer, a prostate cancer, a liver cancer, a colon cancer, a pancreatic cancer, an ovarian cancer, a lymphoma, or a cutaneous T-cell lymphoma, or a melanoma.

[0024] In some embodiments of the first aspect, the method comprises determining baseline levels of the one or more biomarkers or one or more cells in an initial biological sample. In some embodiments, the initial biological sample is obtained on day 1 of thetreatment cycle prior to administration of the one or more immune checkpoint inhibitors and plinabulin. In some embodiments, the method comprises comparing the levels determined in step (iv) with the baseline levels. In some embodiments, the comparing comprises determining a change in the levels of the one or more biomarkers or one or more cells.

[0025] In some embodiments of the first aspect, treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the change in the levels of the one or more biomarkers or one or more cells is below a predetermined threshold. In other embodiments, treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the change in the levels of the one or more biomarkers or one or more cells is above a predetermined threshold.

[0026] In some embodiments, treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the level of pDC expressing CCR7 is greater than about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 or 70 percent or more below the baseline level. In some embodiments, treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the level of pDC expressing CCR7 is less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more above the baseline level.

[0027] In some embodiments, treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the level of pDC expressing CD80 is greater than about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 or 70 percent or more below the baseline level. In some embodiments, treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the level of pDC expressing CD80 is less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more above the baseline level.

[0028] In some embodiments, treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the level of pDC expressing CD83 is greater than about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 or 70 percent or more below the baseline level. In some embodiments, treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the level of pDC expressing CD83 is less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more above the baseline level.

[0029] In a second aspect of the present disclosure, described herein is a method of treating cancer in a subject, the method comprising a treatment cycle comprising the steps of:(i) obtaining a biological sample from the subject;(ii) determining the levels of one or more biomarkers or levels of one or more cells in the biological sample; and(iii) administering plinabulin or a pharmaceutically acceptable salt thereof to the subject if the levels of one or more biomarkers or levels of one or more cells in the biological sample are above or below a threshold level, or if a score determined by mathematically combining levels of two or more biomarkers or two or more cells is above or below a predetermined threshold value.

[0030] In some embodiments of the second aspect, the biological sample is a tumor biopsy. In other embodiments, the biological sample is peripheral blood.

[0031] In some embodiments of the second aspect, the one or more cells are dendritic cells (DC) and their subsets (e.g., myeloid dendritic cells (mDC), plasmacytoid dendritic cells (pDC) conventional dendritic cells (eDC)); or wherein the one or more cells are selected from the group consisting of CD3+ T-cells, CD4+ T-cells CD8+ T-cells, B-cells, IgA+ plasma cells, IgG-i- plasma cells, and IgM-i- plasma cells; or wherein the one or more cells are selected from the group consisting of CD 16+ monocytes, CD 14+ monocytes, natural killer (NK) cells, granzyme K+ T cells, CD8+ TRM cells, CD8+ TEM cells, CD4+ TCM cells, CD4+ TEM cells, CD4+ naive T-cells, naive B-cells, cDCl cells cDC2 cells, and DC3 cells. In some embodiments, the one or more cells are monocyte-derived macrophage cells (MoMac), and interstitial macrophages (IM). In other embodiments, the one or more cells are immune cells from peripheral blood mononuclear cells (PBMCs).

[0032] In some embodiments of the second aspect, determining the score is a GEF- H1 immune activation score. In some embodiments, the GEF-H1 immune activation score is determined using a composite of gene expression levels of one or more genes selected from the group consisting of: Mmpl2, Csfl, Ccl6, Osm, Ccl9, Cxcl3, Clec7a, Hdc, Slcl5a3, Dcstamp, Treml, Cxcl2, Cd300a, Ncf2, Cd80, Lilr4b, Pdcdllg2, Lilrb4a, Mxdl, SpI40. Illa, Ppbp, Gprl71, Ccl7, Illb, Ccl4, Mania, Aqp9, Creb5, Traf4, Nod2, Ptger2, Mefv, Cd6, Reps2, Tlr7, Sirpblb, Fcgr4, P2ry2, Apol8, Lag3, Pdcdl, Cxcrl, Rab27b, Ctla4, Cxcl9, and Fasl.

[0033] In some embodiments of the second aspect, the threshold value of GEF-H1 immune activation score is from about 10 to about 40. In other embodiments, the threshold value of GEF-H1 immune activation score is from about 20 to about 40. In some embodiments,the threshold value of GEF-H1 immune activation score is about 30. In other embodiments, the threshold value of GEF-H1 immune activation score is about 25.

[0034] In some embodiments of the second aspect, the cancer is selected from a breast cancer, a bladder cancer, a glioma, a glioblastoma, a head and neck cancer, a non-small cell lung cancer, a small cell lung cancer, recurrent small cell lung cancer (SCLC), a colorectal cancer, a gastrointestinal stromal tumor, a gastroesophageal carcinoma, a renal cell cancer, a prostate cancer, a liver cancer, a colon cancer, a pancreatic cancer, an ovarian cancer, a lymphoma, or a cutaneous T-cell lymphoma, or a melanoma.

[0035] In some embodiments of the second aspect, the method further includes administration of radiation therapy, an immune checkpoint inhibitor, a chemotherapeutic agent, or a combination thereof, to the subject. In some embodiments, the method further includes administration of radiation therapy and an immune checkpoint inhibitor to the subject.BRIEF DESCRIPTION OF THE DRAWINGS

[0036] FIG. 1 shows mean fluorescence intensity (MFI) levels of surface expressions of MHC-II, CD40, CD80 and CD86 on XS 106 DCs using flow cytometry.

[0037] FIG. 2A shows changes in % pDC that express CCR7, CD80 and CD83 in patients with PD or PR+SD three days after first administration of plinabulin.

[0038] FIG. 2B shows changes in % peripheral blood monocytes represented by classical and inflammatory phenotypes in patients with PD or PR+SD three days after first administration of plinabulin.

[0039] FIG. 3A and 3B shows various cell type proportions of tumor biopsy samples comparing PR+SD subjects versus PD subjects in scRNA-seq analysis.

[0040] FIG. 4A shows a bar plot of normalized baseline GEF-H1 immune activation score for cDCl, cDC2, DC3, and total DC cells in patients with (PR+SD) and without (PD) clinical benefits.

[0041] FIG. 4B shows box plots of GEF-H1 normalized immune scores in DC subsets and total DC cells comparing pre- and post-treatment patients with (PR+SD) and without (PD) clinical benefits.

[0042] FIG. 5A shows single cell RNAseq analysis of tumor biopsies at pre / post treatment for GEF-H1 -dependent immune signature in monocyte-derived macrophages in responding (PR +SD) and non-responding (PD) patients.

[0043] FIG. 5B shows single cell RNAseq analysis of tumor biopsies at pre / post treatment for- ARG 1 (Arginase 1) gene expression in monocyte-derived macrophages in responding (PR +SD) and non-responding (PD) patients.

[0044] FIG. 6A shows a bubble heatmap of GEF-H1 normalized immune scores in 18 immune cell types correlating with clinical response over three time points, spotlighting cell types with statistically significant differences between PR+SD and PD groups.

[0045] FIG. 6B shows a bubble heatmap of GEF-H1 normalized immune scores at the baseline (C1D1) showing cell types correlating with clinical response.

[0046] FIG. 6C shows the bubble heatmap data of GEF-H1 normalized immune scores found in FIG. 6 A in tabular form.

[0047] FIG. 7 depicts a beeswarm plot of single-cell RNA and T-cell receptor (TCR) sequencing analysis of PBMCs.DETAILED DESCRIPTION

[0048] The present disclosure provides method and therapeutic compositions for reversing non-response or enhancing response to immune checkpoint inhibitor therapy for treating, ameliorating, or preventing a cancer or a tumor in a subject using plinabulin. In some embodiments, methods and compositions provided herein are useful in treating, delaying the progression of, preventing relapse of or alleviating a symptom of a cancer or other neoplastic condition using plinabulin. Plinabulin, (3Z,6Z)-3-Benzylidene-6-{ [5-(2-methyl-2-propanyl)- lH-imidazol-4-yl]methylene]-2,5-piperazinedione, is a synthetic analog of the natural compound phenylahistin. Plinabulin can be readily prepared according to methods and procedures detailed in U.S. Pat. Nos. 7,064,201 and 7,919,497, which are incorporated herein by reference in their entireties. Plinabulin can efficiently promote antigen processing and presentation by dendritic cells to effector cells, such as T-cells, and migration of dendritic cells to lymph nodes where tumor- specific antigens are presented by dendritic cells to prime immune effector cells. Exposure of dendritic cells to Plinabulin can induce maturation of dendritic cells and significantly increase their ability to prime T cells.

[0049] Some embodiments disclosed herein include administration of plinabulin, or a pharmaceutically acceptable salt thereof, to a cancer subject, in combination with one or more immune checkpoint inhibitor. In some embodiments, the plinabulin and one or more immune checkpoint inhibitor may be administered in combination with radiation therapy.

[0050] Some embodiments disclosed herein include determining the level of one or more biomarker or one or more cells in a biological sample of a subject after administering plinabulin and one or more immune checkpoint inhibitor and determining whether to discontinue therapy. In some embodiments, the level of particular’ cells expressing one more biomarker may be used to determine whether to discontinue therapy. In some embodiments, the cells are dendritic cells (DC). In some such embodiments, the dendritic cells are myeloid dendritic cells (mDC). In other embodiments, the cells are plasmacytoid dendritic cells (pDC). In still other embodiments, the cells are conventional dendritic cells (cDC). In some embodiments the change in level of one or more biomarker or one or more cells in a biological sample of a subject may be evaluated in relation to a reference level of one or more biomarker or one or more cells in a biological sample of a subject (e.g., in relation to the levels of one or more biomarker or one or more cells in a biological sample of a subject prior to commencing a method of treatment described herein).

[0051] Other embodiments disclosed herein include determining the level of one or more biomarker or one or more cells in a biological sample of a subject before administering plinabulin and one or more immune checkpoint inhibitor, and administering the plinabulin therapy if the level of one or more biomarker or one or more cells in the biological sample are above or below a pre-determined threshold.

[0052] Before the present disclosure is further described, it is to be understood that this invention is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0053] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the invention. The upper and lower limits of thesesmaller ranges may independently be included in the smaller ranges and are also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0054] Methods recited herein may be carried out in any order of the recited events which is logically possible, as well as the recited order of events.

[0055] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are now described.

[0056] All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited.

[0057] It must be noted that as used herein and in the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely,” “only” and the like in connection with the recitation of claim elements, or use of a “negative” limitation.

[0058] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.Definitions

[0059] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents, applications, published applications, and other publications are incorporated by reference in their entirety. In the event that there is a plurality of definitions for a term herein, those in this section prevail unless stated otherwise.

[0060] The term “agent” is used herein to denote a chemical compound, a mixture of chemical compounds, a biological macromolecule, or an extract made from biological materials.

[0061] The term “antagonist” as used herein refers to a compound that can combine with a receptor (e.g., an immune checkpoint receptor) to block a cellular activity. An antagonist may be a ligand that directly binds to the receptor. Alternatively, an antagonist may combine with a receptor indirectly by, for example, (a) forming a complex with another molecule that directly binds to the receptor, or (b) otherwise results in the modification of another compound so that the other compound directly binds to the receptor.

[0062] The term “ameliorate” as used herein refers to any reduction in the extent, severity, frequency, and / or likelihood of a symptom or clinical sign characteristic of a particular condition.

[0063] The term “antibody” or “antibody moiety” is intended to include any polypeptide chain-containing molecular structure with a specific shape that fits to and recognizes an epitope, where one or more non-covalent binding interactions stabilize the complex between the molecular structure and the epitope. Antibodies utilized in the present disclosure may be polyclonal antibodies or monoclonal antibodies. Antibodies also include free antibodies and antigen binding fragments derived therefrom, and conjugates, e.g. pegylated antibodies, drug, radioisotope, or toxin conjugates, and the like. Monoclonal antibodies directed against a specific epitope, or combination of epitopes, will allow for the targeting and / or depletion of cellular populations expressing the marker. Various techniques can be utilized using monoclonal antibodies to screen for cellular populations expressing the marker(s), and include magnetic separation using antibody-coated magnetic beads, “panning” with antibody attached to a solid matrix (i.e., plate), and flow cytometry (See, e.g., U.S. Pat. No. 5,985,660; and Morrison et al. Cell, 96:737-49 (1999)). These techniques allow for the screening of particular populations of cells; in immunohistochemistry of biopsy samples; in detecting the presence of markers shed by cancer cells into the blood and other biologic fluids, and the like. Humanized versions of such antibodies are also within the scope of this disclosure. Humanized antibodies are especially useful for in vivo applications in humans due to their low antigenicity.

[0064] The terms “cancer”, “neoplasm”, and “carcinoma”, are used interchangeably herein to refer to cells which exhibit relatively autonomous growth, so that they exhibit an aberrant growth phenotype characterized by a significant loss of control of cell proliferation. In general, cells of interest for detection or treatment in the present application include precancerous (e.g., benign), malignant, pre-metastatic, metastatic, and non-metastatic cells. Detection of cancerous cells is of particular interest.

[0065] The term “immune checkpoint inhibitor” as used herein refers to a molecule (e.g., small molecule, peptide, polypeptide, protein, antibody, antibody fragment and the like) that acts as an inhibitor (antagonist) of an immune checkpoint pathway. Inhibition of a pathway can include blockade of the pathway through binding to a receptor or signaling molecule that is part of the immune checkpoint pathway.

[0066] The term “polypeptide” is used herein as a generic term to refer to native protein, fragments, or analogs of a polypeptide sequence. Hence, native protein fragments, and analogs are species of the polypeptide genus.

[0067] The term “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. In addition, various adjuvants such as are commonly used in the art may be included. Considerations for the inclusion of various components in pharmaceutical compositions are described, e.g., in Gilman et al. (Eds.) (1990); Goodman and Gilman’s: The Pharmacological Basis of Therapeutics, 8thEd., Pergamon Press, which is incorporated herein by reference in its entirety. The pharmaceutically acceptable excipient can be a monosaccharide or monosaccharide derivative.

[0068] The term “subject” as used herein, means a human or a non-human mammal, e.g., a dog, a cat, a mouse, a rat, a cow, a sheep, a pig, a goat, a non-human primate or a bird, e.g., a chicken, as well as any other vertebrate or invertebrate.

[0069] The term “mammal” is used in its usual biological sense. Thus, it specifically includes, but is not limited to, primates, including simians (chimpanzees, apes,monkeys) and humans, cattle, horses, sheep, goats, swine, rabbits, dogs, cats, rodents, rats, mice, guinea pigs, or the like.

[0070] The terms “effective amount” or a “therapeutically effective amount” as used herein refers to an amount of a therapeutic agent that is effective to relieve, to some extent, or to reduce the likelihood of onset of, one or more of the symptoms of a disease or condition, and can include curing a disease or condition.

[0071] The terms “treat,” “treatment,” or “treating,” as used herein refers to administering a compound or pharmaceutical composition to a subject for prophylactic and / or therapeutic purposes. The term “prophylactic treatment” refers to treating a subject who does not yet exhibit symptoms of a disease or condition, but who is susceptible to, or otherwise at risk of, a particular disease or condition, whereby the treatment reduces the likelihood that the patient will develop the disease or condition. The term “therapeutic treatment” refers to administering treatment to a subject already suffering from a disease or condition.

[0072] The term “biomarker” as used herein to refers to a DNA, RNA, protein, carbohydrate, or glycolipid-based molecular marker, the expression or presence of which in a subject’s or patient’s sample can be detected by standard methods (or methods disclosed herein) and is useful for monitoring the responsiveness or sensitivity of a subject to the combination therapies described herein. Such biomarkers include, but are not limited to, CCR7, CD40, CD80, CD83, CD86, GEF-H1, IL-2, IFNy, IL-6, IL-12p70, IL-12p40, IL-13, IL-17A, IL-23, G-CSF, IL-8 and IFN-P, or a combination thereof. In some embodiments, the biomarker may be Mmpl2, Csfl, Ccl6, Osm, Ccl9, Cxcl3, Clec7a, Hdc, Slcl5a3, Dcstamp, Treml, Cxcl2, Cd300a, Ncf2, Cd80, Lilr4b, Pdcdllg2, Lilrb4a, Mxdl, Spl40. Illa, Ppbp, Gprl71, Ccl7, Il lb, Ccl4, Mania, Aqp9, Creb5, Traf4, Nod2, Ptger2, Mefv, Cd6, Reps2, Tlr7, Sirpblb, Fcgr4, P2ry2, Apol8, Lag3, Pdcdl, Cxcrl, Rab27b, Ctla4, Cxcl9, and Fasl, or a combination thereof. In some embodiments, the level of multiple biomarkers are determined and mathematically combined to determine a combination score. In some embodiments, the combination score may be GEF-H1 immune activation score. Expression of a biomarker may be determined to be higher or lower in a sample obtained from a subject sensitive or responsive to the combination therapies described herein than a pre-determined threshold or a reference level (e.g., the level in a sample previously obtained from the individual at a prior time).

[0073] As used herein, the term “chemotherapeutic agent” refers to an agent that reduces, prevents, mitigates, limits, and / or delays the growth of metastases or neoplasms, or kills neoplastic cells directly by necrosis or apoptosis of neoplasms or any other mechanism, or that can be otherwise used, in a pharmaceutically-effective amount, to reduce, prevent, mitigate, limit, and / or delay the growth of metastases or neoplasms in a subject with neoplastic disease. Chemotherapeutic agents include but are not limited to, for example, fluoropyrimidines; pyrimidine nucleosides; purine nucleosides; anti-folates, platinum-based agents; anthracyclines / anthracenediones; epipodophyllotoxins; camptothecins; hormones; hormonal complexes; antihormonals; enzymes, proteins, peptides and polyclonal and / or monoclonal antibodies; vinca alkaloids; taxanes; epothilones; antimicrotubule agents; alkylating agents; antimetabolites; topoisomerase inhibitors; antivirals; and various other cytotoxic and cytostatic agents.Immune Checkpoint Inhibitors

[0074] In some embodiments, one or more immune checkpoint inhibitor may be co-administered with plinabulin. A review describing immune checkpoint pathways and the blockade of such pathways with immune checkpoint inhibitor compounds is provided by Pardoll in Nature Reviews Cancer (April, 2012), pages 252-264, which is incorporated herein by reference in its entirety. Immune check point inhibitor compounds display anti-tumor activity by blocking one or more of the endogenous immune checkpoint pathways that downregulate an anti-tumor immune response. The inhibition or blockade of an immune checkpoint pathway typically involves inhibiting a checkpoint receptor and ligand interaction with an immune checkpoint inhibitor compound to reduce or eliminate the down regulation signal and resulting diminishment of the anti-tumor response.

[0075] In some embodiments of the present disclosure, the immune checkpoint inhibitor compound inhibits the signaling interaction between an immune checkpoint receptor and the corresponding ligand of the immune checkpoint receptor. The immune checkpoint inhibitor compound can act by blocking activation of the immune checkpoint pathway by inhibition (antagonism) of an immune checkpoint receptor (some examples of receptors include CTLA-4, PD-1, LAG-3, TIM-3, BTLA, and KIR) or by inhibition of a ligand of an immune checkpoint receptor (some examples of ligands include PD-L1 and PD-L2). In such embodiments, the effect of the immune checkpoint inhibitor compound is to reduce oreliminate down regulation of certain aspects of the immune system anti-tumor response in the tumor microenvironment.

[0076] The Programmed Death 1 (PD-1) protein is an inhibitory member of the extended CD28 / CTLA-4 family of T cell regulators (Okazaki et al. (2002) Curr Opin Immunol 14: 391779-82; Bennett et al. (2003) J. Immunol. 170:711-8; which are incorporated herein by reference in their entirety). Other members of the CD28 family include CD28, CTLA-4, ICOS and BTLA. PD-1 is suggested to exist as a monomer, lacking the unpaired cysteine residue characteristic of other CD28 family members. PD-1 is expressed on activated B cells, T cells, and monocytes.

[0077] The PD-1 gene encodes a 55 kDa type I transmembrane protein (Agata et al. (1996) Int Immunol. 8:765-72, which is incorporated herein by reference in its entirety). Although structurally similar to CTLA-4, PD-1 lacks the MYPPY motif that is important for B7-1 and B7-2 binding. Two ligands for PD-1 have been identified, PD-L1 (B7-H1) and PD- L2 (B7-DC), that have been shown to downregulate T cell activation upon binding to PD-1 (Freeman et al. (2000) J. Exp. Med. 192:1027-34; Carteret al. (2002) Eur. J. Immunol. 32:634- 43; which are incorporated herein by reference in their entirety). Both PD-L1 and PD-L2 are B7 homologs that bind to PD-1, but do not bind to other CD28 family members. PD-L1 is abundant in a variety of human cancers (Dong et al. (2002) Nat. Med. 8:787-9, which is incorporated herein by reference in its entirety).

[0078] PD-1 is known as an immunoinhibitory protein that negatively regulates TCR signals (Ishida, Y. et al. (1992) EMBO J. 11:3887-3895; Blank, C. et al. (Epub 2006 Dec. 29) Immunol. Immunother. 56(5):739-745; which are incorporated herein by reference in their entirety). The interaction between PD-1 and PD-L1 can act as an immune checkpoint, which can lead to, e.g., a decrease in tumor infiltrating lymphocytes, a decrease in T-cell receptor mediated proliferation, and / or immune evasion by cancerous cells (Dong et al. (2003) J. Mol. Med. 81:281-7; Blank et al. (2005) Cancer Immunol. Immunother. 54:307-314; Konishi et al. (2004) Clin. Cancer Res. 10:5094-100; which are incorporated herein by reference in their entirety). Immune suppression can be reversed by inhibiting the local interaction of PD- 1 with PD-L1 or PD-L2; the effect is additive when the interaction of PD-1 with PD-L2 is blocked as well (Iwai et al. (2002) Proc. Nat’l. Acad. Sci. USA 99:12293-7; Brown et al. (2003) J. Immunol. 170:1257-66; which are incorporated herein by reference in their entirety).

[0079] The immune checkpoint receptor cytotoxic T-lymphocyte associated antigen 4 (CTLA-4) is expressed on T-cells and is involved in signaling pathways that reduce the level of T-cell activation. It is believed that CTLA-4 can downregulate T-cell activation through competitive binding and sequestration of CD80 and CD86. In addition, CTLA-4 has been shown to be involved in enhancing the immunosuppressive activity of TRegcells.

[0080] The immune checkpoint receptor programmed death 1 (PD-l) is expressed by activated T-cells upon extended exposure to antigen. Engagement of PD-l with its known binding ligands, PD-L1 and PD-L2, occurs primarily within the tumor microenvironment and results in downregulation of anti-tumor specific T-cell responses. Both PD-L1 and PD-L2 are known to be expressed on tumor cells. The expression of PD-L1 and PD-L2 on tumors has been correlated with decreased survival outcomes.

[0081] The immune checkpoint receptor T cell membrane protein 3 (TIM-3) is expressed on Thl and Tel cells, but not other T-cells. Interaction of TIM-3 with its ligand, galectin-9, produces a Thl cell death signal. TIM-3 has been reported to play a role in maintaining T-cell exhaustion and blockade of TIM-3 has been shown to restore activity to exhausted T-cells.

[0082] The immune checkpoint receptor B- and T-lymphocyte attenuator (BTLA) receptor is expressed on both resting and activated B-cells and T-cells. Activation of BTLA when combined with its ligand HVEM (herpes virus entry mediator) results in downregulation of both T-cell activation and proliferation. HVEM is expressed by certain tumors (e.g., melanoma) and tumor- associated endothelial cells.

[0083] The immune checkpoint receptors known as killer cell immunoglobulin- like receptors (KIR) are a polymorphic family of receptors expressed on NK cells and some T- cells and function as regulators of immune tolerance associated with natural killer (NK) cells. Blocking certain KIR receptors with inhibitor compounds can facilitate the destruction of tumors through the increased activity of NK cells.

[0084] In some embodiments of the present disclosure, the immune checkpoint inhibitor compound is a small organic molecule (molecular weight less than 1000 daltons), a peptide, a polypeptide, a protein, an antibody, an antibody fragment, or an antibody derivative. In some embodiments, the immune checkpoint inhibitor compound is an antibody. In someembodiments, the antibody is a monoclonal antibody, specifically a human or a humanized monoclonal antibody.

[0085] Monoclonal antibodies, antibody fragments, and antibody derivatives for blocking immune checkpoint pathways can be prepared by any of several methods known to those of ordinary skill in the art, including but not limited to, somatic cell hybridization techniques and hybridoma, methods. Hybridoma generation is described in Antibodies, A Laboratory Manual, Harlow and Lane, 1988, Cold Spring Harbor Publications, New York, which is incorporated herein by reference in its entirety. Human monoclonal antibodies can be identified and isolated by screening phage display libraries of human immunoglobulin genes by methods described for example in U.S. Pat. Nos. 5,223,409, 5,403,484, 5,571,698, 6,582,915, and 6,593,081, which are incorporated herein by reference in their entirety. Monoclonal antibodies can be prepared using the general methods described in U.S. Pat. No. 6,331,415 (Cabilly), which is incorporated herein by reference in its entirety.

[0086] As an example, human monoclonal antibodies can be prepared using a XenoMouse™ (Abgenix, Freemont, Calif.) or hybridomas of B cells from a XenoMouse. A XenoMouse is a murine host having functional human immunoglobulin genes as described in U.S. Pat. No. 6,162,963 (Kucherlapati), which is incorporated herein by reference in its entirety.

[0087] Methods for the preparation and use of immune checkpoint antibodies are described in the following illustrative publications. The preparation and therapeutic uses of anti-CTLA-4 antibodies are described in U.S. Pat. No. 7,229,628 (Allison), U.S. Pat. No. 7,311,910 (Linsley), and U.S. Pat. No. 8,017,144 (Korman), which are incorporated herein by reference in their entirety. The preparation and therapeutic uses of anti-PD-1 antibodies are described in U.S. Pat. No. 8,008,449 (Korman) and U.S. Patent Application No. 2011 / 0271358 (Freeman), which are incorporated herein by reference in their entirety. The preparation and therapeutic uses of anti-PD-Ll antibodies are described in U.S. Pat. No. 7,943,743 (Korman), which is incorporated herein by reference in its entirety. The preparation and therapeutic uses of anti-TIM-3 antibodies are described in U.S. Pat. No. 8,101,176 (Kuchroo) and U.S. Pat. No. 8,552,156 (Tagayanagi), which are incorporated herein by reference in their entirety. The preparation and therapeutic uses of anti-LAG-3 antibodies are described in U.S. Patent Application No. 2011 / 0150892 (Thudium) and International Publication NumberWO2014 / 008218 (Lonberg), which are incorporated herein by reference in their entirety. The preparation and therapeutic uses of anti-KIR antibodies arc described in U.S. Pat. No. 8,119,775 (Moretta), which is incorporated herein by reference in its entirety. The preparation of antibodies that block BTLA regulated inhibitory pathways (anti-BTLA antibodies) are described in U.S. Pat. No. 8,563,694 (Mataraza), which is incorporated herein by reference in its entirety.

[0088] In some embodiments, the one or more immune checkpoint inhibitor is an inhibitor of PD-1, PD-L1, or CTLA-4. In some embodiments, the immune checkpoint inhibitor is a PD- 1 inhibitor. In some embodiments, the immune checkpoint inhibitor is a binding ligand of PD-L1. In some embodiments, the immune checkpoint inhibitor is a PD-L1 inhibitor. In some embodiments, the immune checkpoint inhibitor is a CTLA-4 inhibitor.

[0089] In some embodiments, the one or more immune checkpoint inhibitor as described herein includes a first immune checkpoint inhibitor and a second immune checkpoint inhibitor, wherein the first immune checkpoint inhibitor is different from the second immune checkpoint inhibitor. In some embodiments, the first and the second immune checkpoint inhibitor are independently an inhibitor of PD-1, PD-L1 or CTLA-4. In some embodiments, the first immune checkpoint inhibitor is a PD-1 inhibitor, and the second immune checkpoint inhibitor is a CTLA-4 inhibitor.

[0090] In some embodiments, the immune checkpoint inhibitor is Pembrolizumab (SEQ ID NO:1), nivolumab (SEQ ID NO:2), cemiplimab (SEQ ID NO:3), atezolizumab (SEQ ID NO:4), avelumab (SEQ ID NO:5), pembrolizumab (SEQ ID NO:6), pidilizumab (SEQ ID NO:7), ipilimumab (SEQ ID NO:8), BMS 936559 (SEQ ID NO:9), durvalumab (SEQ ID NO: 10), camrelizumab (SEQ ID NO: 10), dostarlimab (SEQ ID NO: 11), tislelizumab (SEQ ID NO: 12), sintilimab (SEQ ID NO: 13), toripalimab (SEQ ID NO: 14), or any combinations thereof. In some embodiments, the one or more immune checkpoint inhibitor may include an anti-PD-1 HuMAbs can be selected from 17D8, 2D3, 4H1, 5C4 (also referred to herein as nivolumab), 4A1 1, 7D3 and 5F4, all of which are described in U.S. Pat. No. 8,008,449, which is incorporated herein by reference in its entirety. In some embodiments, the anti-PD-1 HuMAbs can be selected from 3G10, 12A4 (also referred to herein as BMS-936559), 10A5, 5F8, 10H10, IB 12, 7H1, 1 1E6, 12B7, and 13G4, all of which are described in U.S. Pat. No. 7,943,743, which is incorporated herein by reference in its entirety.

[0091] In some embodiments, the one or more immune checkpoint inhibitor may be incorporated in a pharmaceutically acceptable formulation. In some embodiments, the one or more immune checkpoint inhibitor is incorporated in a pharmaceutically acceptable aqueous formulation. Examples of acceptable aqueous formulations include isotonic buffered and pH 4.5-8 adjusted saline solutions such as Lactated Ringer’s Solution and the like.

[0092] In some embodiments, the immune checkpoint inhibitor compound is incorporated in a pharmaceutically acceptable liposome formulation, wherein the formulation is a passive or targeted liposome formulation. Examples of methods for the preparation of suitable liposome formulations of antibodies are described U.S. Pat. No. 5,399,331 (Loughrey), U.S. Pat. No. 8,304,565 (Wu) and U.S. Pat. No. 7,780,882 (Chang), which are incorporated herein by reference in their entirety.

[0093] In some embodiments, the one or more immune checkpoint inhibitor may be an antibody. In some embodiments, the antibody is a dry, lyophilized solid that is reconstituted with an aqueous reconstitution solvent prior to use. In some embodiments, the antibody is incorporated in a pharmaceutically acceptable formulation and the pharmaceutically acceptable formulation is injected directly into a tumor. In some embodiments, the immune checkpoint inhibitor antibody is incorporated in a pharmaceutically acceptable formulation and the pharmaceutically acceptable formulation is injected into the peritumoral region surrounding a tumor. The peritumoral region may contain antitumor immune cells. In some embodiments, the antibody is incorporated in a pharmaceutically acceptable formulation and the pharmaceutically acceptable formulation is administered by intravenous injection or infusion. In some embodiments, the immune checkpoint inhibitor antibody is incorporated in a pharmaceutically acceptable formulation and the pharmaceutically acceptable formulation is administered by subcutaneous injection or intradermal injection. In some embodiments, the antibody is incorporated in a pharmaceutically acceptable formulation and the pharmaceutically acceptable formulation is administered by intraperitoneal injection or lavage.

[0094] The precise amount of immune checkpoint inhibitor compound incorporated in a particular method or therapeutic combination of the disclosure may vary according to factors known in art such as for example, the physical and clinical status of the subject, the method of administration, the content of the formulation, the physical and chemicalnature of the immune checkpoint inhibitor compound, the intended dosing regimen or sequence. Those of ordinary skill in the art, however, can readily determine the appropriate amount with due consideration of such factors.Chemotherapeutic Agents

[0095] In some embodiments, the plinabulin and one or more immune checkpoint inhibitor may be administered in combination with an additional chemotherapeutic agent. In some embodiments, an additional chemotherapeutic agent can be selected from the group consisting of Abiraterone, Acetate, Abitrexate (Methotrexate), Abraxane (Paclitaxel Albumin- stabilized Nanoparticle Formulation), ABVD, ABVE, ABVE-PC , AC, AC-T, ADE, Ado- Trastuzumab Emtansine ,Adriamycin (Doxorubicin Hydrochloride) , Afatinib Dimaleate, Afinitor (Everolimus), Akynzeo (Netupitant and Palonosetron Hydrochloride), Aldara (Imiquimod), Aldesleukin, Alecensa (Alectinib), Alectinib, Alemtuzumab, Alimta (Pemetrexed Disodium), Aloxi (Palonosetron Hydrochloride), Ambochlorin (Chlorambucil), Amboclorin (Chlorambucil), Aminolevulinic Acid, \ Anastrozole, Aprepitant, Aredia (Pamidronate Disodium), Arimidex (Anastrozole), Aromasin (Exemestane), Arranon (Nelarabine), Arsenic Trioxide, Arzerra (Ofatumumab), Asparaginase Erwinia chrysanthemi , Avastin (Bevacizumab), Axitinib, Azacitidine, BEACOPP, Becenum (Carmustine), Beleodaq (Belinostat), Belinostat, Bendamustine Hydrochloride, BEP, Bevacizumab, Bexarotene, Bexxar (Tositumomab and Iodine I 131 Tositumomab), Bicalutamide, BiCNU (Carmustine), Bleomycin, Blinatumomab, Blincyto (Blinatumomab), Bortezomib, Bosulif (Bosutinib), Bosutinib, Brentuximab Vedotin, Busulfan, Cabazitaxel, Cabozantinib-S-Malate, CAF, Campath (Alemtuzumab), Camptosar (Irinotecan Hydrochloride), Capecitabine, CAPOX, Carac (Fluorouracil — Topical), Carboplatin, CARBOPLATIN-TAXOL, Carfilzomib, Carmubris (Carmustine), Carmustine, Carmustine Implant, Casodex (Bicalutamide), CeeNU (Lomustine), Ceritinib, Cerubidine (Daunorubicin Hydrochloride), Cervarix (Recombinant HPV Bivalent Vaccine), Cetuximab, Chlorambucil, CHLORAMBUCIL-PREDNISONE, CHOP, Cisplatin, Clafen (Cyclophosphamide), Clofarabine, Clofarex (Clofarabine), Clolar (Clofarabine), CMF, Cobimetinib, Cometriq (Cabozantinib-S-Malate), COPDAC, COPP, COPP-ABV, Cosmegen (Dactinomycin), Cotellic (Cobimetinib), Crizotinib, CVP, Cyclophosphamide, Cyfos (Ifosfamide), Cyramza (Ramucirumab), Cytarabine, Cytarabine Liposome, Cytosar-U (Cytarabine), Cytoxan (Cyclophosphamide), Dabrafenib, Dacarbazine,Dacogen (Decitabine), Dactinomycin, Daratumumab, Darzalex (Daratumumab), Dasatinib, Daunorubicin Hydrochloride, Decitabine, Dcgarclix, Dcnilcukin Diftitox, Dcnosumab, DepoCyt (Cytarabine Liposome), Dexamethasone, Dexrazoxane Hydrochloride, Dinutuximab, Docetaxel, Doxil (Doxorubicin Hydrochloride Liposome), Doxorubicin Hydrochloride, Doxorubicin Hydrochloride Liposome, Dox-SL (Doxorubicin Hydrochloride Liposome), Efudex (Fluorouracil — Topical), Elitek (Rasburicase), Ellence (Epirubicin Hydrochloride), Elotuzumab, Eloxatin (Oxaliplatin), Eltrombopag Olamine, Emend (Aprepitant), Empliciti (Elotuzumab), Enzalutamide, Epirubicin Hydrochloride, EPOCH, Erbitux (Cetuximab), Eribulin Mesylate, Erivedge (Vismodegib), Erlotinib Hydrochloride, Erwinaze (Asparaginase Erwinia chrysanthemi), Etopophos (Etoposide Phosphate), Etoposide, Etoposide Phosphate, Evacet (Doxorubicin Hydrochloride Liposome), Everolimus, Evista (Raloxifene Hydrochloride), Exemestane, 5-FU (Fluorouracil Injection), 5-FU (Fluorouracil — Topical), Fareston (Toremifene), Farydak (Panobinostat), Faslodex (Fulvestrant), FEC, Femara (Letrozole), Filgrastim, Fludara (Fludarabinc Phosphate), Fludarabine Phosphate, Fluoroplex (Fluorouracil — Topical), Fluorouracil Injection, Fluorouracil — Topical, Flutamide, Folex (Methotrexate), Folex PFS (Methotrexate), FOLFIRI, FOLFIRI- BEVACIZUMAB, FOLFIRI-CETUXIMAB, FOLFIRINOX, FOLFOX, Folotyn (Pralatrexate), FU-LV, Fulvestrant, Gardasil (Recombinant HPV Quadrivalent Vaccine), Gardasil 9 (Recombinant HPV Nonavalent Vaccine), Gazyva (Obinutuzumab), Gefitinib, Gemcitabine Hydrochloride, GEMCITABINE-CISPLATIN, GEMCITABINEOXALIPLATIN ,Gemtuzumab Ozogamicin, Gemzar (Gemcitabine Hydrochloride), Gilotrif (Afatinib Dimaleate), Gleevec (Imatinib Mesylate), Gliadel (Carmustine Implant), Gliadel wafer (Carmustine Implant), Glucarpidase, Goserelin Acetate, Halaven (Eribulin Mesylate), Herceptin (Trastuzumab), HPV Bivalent Vaccine, Recombinant, HPV Nonavalent Vaccine, Recombinant, HPV Quadrivalent Vaccine, Recombinant, Hycamtin (Topotecan Hydrochloride), Hyper-CVAD, Ibrance (Palbociclib), Ibritumomab Tiuxetan, Ibrutinib, ICE, Iclusig (Ponatinib Hydrochloride), Idamycin (Idarubicin Hydrochloride), Idelalisib, Ifex (Ifosfamide), Ifosfamide, IL-2 (Aldesleukin), Imatinib Mesylate, Imbruvica (Ibrutinib), Imiquimod, Imlygic (Talimogene Laherparepvec), Inlyta (Axitinib), Interferon Alfa-2b, Recombinant, Interleukin-2 (Aldesleukin), Intron A (Recombinant Interferon Alfa-2b), Iodine 1 131 Tositumomab and Tositumomab, Iressa (Gefitinib), Irinotecan Hydrochloride, IrinotecanHydrochloride Liposome, Istodax (Romidepsin), Ixabepilone, Ixazomib Citrate, Ixempra (Ixabcpilonc), Jakafi (Ruxolitinib Phosphate), Jcvtana (Cabazitaxcl), Kadcyla (Ado- Trastuzumab Emtansine), Keoxifene (Raloxifene Hydrochloride), Kepivance (Palifermin), Kyprolis (Carfilzomib), Lanreotide Acetate, Lapatinib Ditosylate, Lenalidomide, Lenvatinib Mesylate, Lenvima (Lenvatinib Mesylate), Letrozole, Leucovorin Calcium, Leukeran (Chlorambucil), Leuprolide Acetate, Levulan (Aminolevulinic Acid), Linfolizin (Chlorambucil), LipoDox (Doxorubicin Hydrochloride Liposome), Lomustine, Lonsurf (Trifluridine and Tipiracil Hydrochloride), Lupron (Leuprolide Acetate) ,Lupron Depot (Leuprolide Acetate), Lupron Depot-Ped (Leuprolide Acetate), Lupron Depot-3 Month (Leuprolide Acetate), Lupron Depot-4 Month (Leuprolide Acetate), Lynparza (Olaparib), Marqibo (Vincristine Sulfate Liposome), Matulane (Procarbazine Hydrochloride), Mechlorethamine Hydrochloride, Megace (Megestrol Acetate), Megestrol Acetate, Mekinist (Trametinib), Mercaptopurine, Mesna, Mesnex (Mesna), Methazolastone (Temozolomide), Methotrexate, Methotrexate LPL (Methotrexate), Mexate (Methotrexate), Mexate-AQ (Methotrexate), Mitomycin C, Mitoxantrone Hydrochloride, Mitozytrex (Mitomycin C), MOPP,Mozobil (Plerixafor), Mustargen (Mechlorethamine Hydrochloride) „Mutamycin (Mitomycin C), Myleran (Busulfan), Mylosar (Azacitidine), Mylotarg (Gemtuzumab Ozogamicin), Nanoparticle Paclitaxel (Paclitaxel Albumin-stabilized Nanoparticle Lormulation), Navelbine (Vinorelbine Tartrate), Necitumumab, Nelarabine, Neosar (Cyclophosphamide), Netupitant and Palonosetron Hydrochloride, Neupogen (Lilgrastim), Nexavar (Sorafenib Tosylate) , Nilotinib, Ninlaro (Ixazomib Citrate), Nolvadex (Tamoxifen Citrate), Nplate (Romiplostim), Obinutuzumab, Odomzo (Sonidegib), OEPA, Ofatumumab, OPP, Olaparib, Omacetaxine Mepesuccinate, Oncaspar (Pegaspargase), Ondansetron Hydrochloride, Onivyde (Irinotecan Hydrochloride Liposome), Ontak (Denileukin Diftitox), OPPA , Osimertinib, Oxaliplatin, Paclitaxel, Paclitaxel Albumin-stabilized Nanoparticle Lormulation, PAD, Palbociclib, Palifermin, Palonosetron Hydrochloride, Palonosetron Hydrochloride and Netupitant, Pamidronate Disodium, Panitumumab, Panobinostat, Paraplat (Carboplatin), Paraplatin (Carboplatin), Pazopanib Hydrochloride, PCV , Pegaspargase, Peginterferon Alfa-2b, PEG-Intron (Peginterferon Alfa- 2b), Pemetrexed Disodium Perjeta (Pertuzumab), Pertuzumab, Platinol (Cisplatin), Platinol-AQ (Cisplatin), Plerixafor, Pomalidomide, Pomalyst (Pomalidomide), Ponatinib Hydrochloride, Portrazza(Necitumumab), Pralatrexate, Prednisone, Procarbazine Hydrochloride, Proleukin (Aldesleukin), Prolia (Dcnosumab), Promacta (Eltrombopag Olaminc), Provcngc (Sipulcuccl- T), Purinethol (Mercaptopurine), Purixan (Mercaptopurine), Radium 223 Dichloride, Raloxifene Hydrochloride, Ramucirumab, Rasburicase, R-CHOP, R-CVP, Recombinant Human Papillomavirus (HPV) Bivalent Vaccine, Recombinant Human Papillomavirus (HPV) Nonavalent Vaccine, Recombinant Human Papillomavirus (HPV) Quadrivalent Vaccine, Recombinant Interferon Alfa-2b, Regorafenib, R-EPOCH, Revlimid (Lenalidomide), Rheumatrex (Methotrexate), Rituximab, Rolapitant Hydrochloride, Romidepsin, Romiplostim, Rubidomycin (Daunorubicin Hydrochloride), Ruxolitinib Phosphate, Sclerosol Intrapleural Aerosol (Talc),Siltuximab, SipuleuceLT, Somatuline Depot (Lanreotide Acetate), Sonidegib, Sorafenib Tosylate, Sprycel (Dasatinib), STANFORD V, Sterile Talc Powder (Talc), Steritalc (Talc), Stivarga (Regorafenib), Sunitinib Malate, Sutent (Sunitinib Malate), Sylatron (Peginterferon Alfa-2b), Sylvant (Siltuximab), Synovir (Thalidomide), Synribo (Omacetaxine Mepesuccinate), Tabloid (Thioguanine), TAC, Tafinlar (Dabrafenib), Tagrisso (Osimertinib), Talc, Talimogene Laherparepvec, Tamoxifen Citrate, Tarabine PFS (Cytarabine), Tarceva (Erlotinib Hydrochloride), Targretin (Bexarotene), Tasigna (Nilotinib), Taxol (Paclitaxel), Taxotere (Docetaxel), Temodar (Temozolomide), Temozolomide, Temsirolimus, Thalidomide, Thioguanine, Thiotepa, Tolak (Fluorouracil — Topical), Toposar (Etoposide), Topotecan Hydrochloride, Toremifene, Torisel (Temsirolimus), Tositumomab and Iodine I 131, Tositumomab, Totect (Dexrazoxane Hydrochloride), TPF, Trabectedin, Trametinib, Trastuzumab, Treanda (Bendamustine Hydrochloride) , Trifluridine and Tipiracil Hydrochloride, Trisenox (Arsenic Trioxide), Tykerb (Lapatinib Ditosylate), Unituxin (Dinutuximab), Uridine Triacetate, VAC, Vandetanib, VAMP, Varubi (Rolapitant Hydrochloride), Vectibix (Panitumumab), VelP, Velban (Vinblastine Sulfate), Velcade (Bortezomib), Velsar (Vinblastine Sulfate), Vemurafenib, VePesid (Etoposide), Viadur (Leuprolide Acetate), Vidaza (Azacitidine), Vinblastine Sulfate, Vincasar PFS (Vincristine Sulfate), Vincristine Sulfate, Vincristine Sulfate Liposome, Vinorelbine Tartrate, VIP, Vismodegib, Vistogard (Uridine Triacetate), Voraxaze (Glucarpidase), Vorinostat, Votrient (Pazopanib Hydrochloride), Wellcovorin (Leucovorin Calcium), Xalkori (Crizotinib), Xeloda (Capecitabine), XELIRI,XELOX, Xgeva (Denosumab), Xofigo (Radium 223 Dichloride), Xtandi (Enzalutamide), Yervoy (Ipilimumab), Yondelis (Trabectedin), Zaltrap (Ziv-Aflibercept), Zarxio (Filgrastim), Zelboraf (Vemurafenib), Zevalin (Ibritumomab Tiuxetan), Zinccard (Dcxrazoxanc Hydrochloride), Ziv-Aflibcrccpt, Zofran (Ondansetron Hydrochloride), Zoladex (Goserelin Acetate), Zoledronic Acid, Zolinza (Vorinostat), Zometa (Zoledronic Acid), Zydelig (Idelalisib), Zykadia (Ceritinib), and Zytiga (Abiraterone Acetate).

[0096] In some embodiments, the additional chemotherapeutic agent is docetaxel.

[0097] The precise amount of additional chemotherapeutic agent incorporated in a particular method of the disclosure may vary according to factors known in art such as for example, the physical and clinical status of the subject, the method of administration, the content of the formulation, the physical and chemical nature of the additional chemotherapeutic agent, the intended dosing regimen or sequence. Those of ordinary skill in the art, however, can readily determine the appropriate amount with due consideration of such factors.Radiation Therapy

[0098] In some embodiments, the combination of plinabulin and one or more immune checkpoint inhibitor is co-administered with radiation. In some embodiments, the radiation may be selected from external beam radiation therapy or internal radiation therapy. In some embodiments, the external beam radiation therapy may be selected from three- dimensional conformal radiation therapy (3D-CRT), intensity modulated radiation therapy (IMRT), proton beam therapy, image-guided radiation therapy (IGRT), Stereotactic radiation therapy (SRT), or a combination thereof. In some embodiments, the radiation may be selected from intraoperative radiation therapy (IORT), systemic radiation therapy, radioimmunotherapy, radiosensitizers, radioprotectors, or a combination thereof.Use and Method of Treatment

[0099] In some aspects, provided herein is a method of treating cancer in a subject, the method comprising a treatment cycle comprising the steps of: (i) administering one or more immune checkpoint inhibitor to the subject; (ii) administering plinabulin or a pharmaceutically acceptable salt thereof to the subject; (iii) obtaining a biological sample from the subject; and (iv) determining the levels of one or more biomarkers or levels of one or more cells in the biological sample.

[0100] The treatment cycles disclosed herein being on day 1 and end on the last day of the cycle. For example, three week treatment cycles begin on day 1 and end on day 21, while four week treatment cycles begin on day 1 and end on day 28.

[0101] In some embodiments, the biological sample may be a blood sample. In other embodiments, the biological sample may be a tumor biopsy. In some embodiments, the biological sample may be obtained from the subject before commencement of the treatment cycle, e.g., on day 1 of the treatment cycle or before day 1 of the treatment cycle. In some embodiments, the biological sample may be obtained during the treatment cycle. For example, in some embodiments, the biological sample may be obtained from the subject on day 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, and / or 28 of the treatment cycle.

[0102] In some embodiments, the treatment cycle may be 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 days, or longer. In some embodiments, the treatment cycle may be 14 days (three weeks or “Q2W”). In some embodiments, the treatment cycle may be 21 days (three weeks or “Q3W”). In some embodiments, the treatment cycle may be 28 days (four weeks or “Q4W”). In some embodiments, the treatment cycle may be repeated 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more times. In some embodiments, the steps of obtaining a biological sample from the subject; and determining the levels of one or more biomarkers or levels of one or more cells in the biological sample may be repeated during each treatment cycle.

[0103] In some embodiments, the treatment cycle comprises administering the one or more immune checkpoint inhibitor and plinabulin to the subject on day 1 of the treatment cycle and obtaining a biological sample on day 4 of the treatment cycle. In some such embodiments, a biological sample may be obtained from the subject on day 1 of the treatment cycle prior to administration of the one or more immune checkpoint inhibitor and plinabulin. In other such embodiments, a biological sample may be obtained from the subject prior to day 1 of the treatment cycle.

[0104] In some embodiments, the one or more biomarker is CCR7, CD40, CD80, CD83, CD86, GEF-H1, IL-2, IFNy, IL-6, IL-12p70, IL-12p40, IL-13, IL-17A, IL-23, G-CSF, PD-L1, IL-8 and IFN-p, or a combination thereof. In some embodiments, the one or more biomarker is CCR7, CD40, CD80, CD83, CD86, or a combination thereof. In some embodiments, the biomarker is Mmpl2, Csfl, Ccl6, Osm, Ccl9, Cxcl3, Clec7a, Hdc, Slcl5a3, Dcstamp, Treml, Cxcl2, Cd300a, Ncf2, Cd80, Lilr4b, Pdcdllg2, Lilrb4a, Mxdl, Spl40. Illa, Ppbp, Gprl71, Ccl7, Illb, Ccl4, Mania, Aqp9, Creb5, Traf4, Nod2, Ptger2, Mefv, Cd6, Reps2,Tlr7, Sirpblb, Fcgr4, P2ry2, Apol8, Lag3, Pdcdl , Cxcrl , Rab27b, Ctla4, Cxcl9, and Fasl, or a combination thereof. In some embodiments, the level of multiple biomarkers arc determined and mathematically combined to determine a combination score. In some embodiments, the combination score is a GEF-H1 immune activation score. In some embodiments, the GEF-H1 immune activation score is determined using a composite of gene expression levels of one or more genes selected from the group consisting of: Mmpl2, Csfl, Ccl6, Osm, Ccl9, Cxcl3, Clec7a, Hdc, Slcl5a3, Dcstamp, Treml, Cxcl2, Cd300a, Ncf2, Cd80, Lilr4b, Pdcdllg2, Lilrb4a, Mxdl, Spl40. Illa, Ppbp, Gprl71, Ccl7, Illb, Ccl4, Mania, Aqp9, Creb5, Traf4, Nod2, Ptger2, Mefv, Cd6, Reps2, Tlr7, Sirpblb, Fcgr4, P2ry2, Apol8, Lag3, Pdcdl, Cxcrl, Rab27b, Ctla4, Cxcl9, and Fasl. In some embodiments, the one or more biomarker includes CCR7. In some embodiments, the one or more biomarker includes CD40. In some embodiments, the one or more biomarker includes CD80. In some embodiments, the one or more biomarker includes CD83. In some embodiments, the one or more biomarker includes CD86.

[0105] In some embodiments, the level of one or more cells in a biological sample may be determined using techniques known in the art. In some embodiments, the one or more cells may be dendritic cells. In some such embodiments, the dendritic cells are myeloid dendritic cells (mDC). In other embodiments, the cells are plasmacytoid dendritic cells (pDC). In still other embodiments, the cells are conventional dendritic cells (cDC). In some embodiments, the levels of one or more dendritic cells in a biological sample expressing one or more biomarkers. In some embodiments, the one or more biomarker is CCR7, CD40, CD80, CD83, CD86, GEF-H1, IL-2, IFNy, IL-6, IL-12p70, IL-12p40, IL-13, IL-17A, IL-23, G-CSF, IL-8 and IFN-0, or a combination thereof. In some embodiments, the one or more biomarker is CCR7, CD40, CD80, CD83, CD86, or a combination thereof. In some embodiments, the biomarker is Mmpl2, Csfl, Ccl6, Osm, Ccl9, Cxcl3, Clec7a, Hdc, Slcl5a3, Dcstamp, Treml, Cxcl2, Cd300a, Ncf2, Cd80, Lilr4b, Pdcdllg2, Lilrb4a, Mxdl, Spl40. Illa, Ppbp, Gprl71, Ccl7, Illb, Ccl4, Mania, Aqp9, Creb5, Traf4, Nod2, Ptger2, Mefv, Cd6, Reps2, Tlr7, Sirpblb, Fcgr4, P2ry2, Apol8, Lag3, Pdcdl, Cxcrl, Rab27b, Ctla4, Cxcl9, and Fasl, or a combination thereof. In some embodiments, the levels of one or more cells (e.g., dendritic cells) having an GEF-H1 immune activation score above a pre-determined threshold is determined. In some embodiments, the one or more biomarker includes CCR7. In some embodiments, the one ormore biomarker includes CD40. In some embodiments, the one or more biomarker includes CD80. In some embodiments, the one or more biomarkcr includes CD83. In some embodiments, the one or more biomarker includes CD86. In some embodiments wherein the biological sample is a tumor biopsy, the one or biomarkers includes PD-L1.

[0106] The level of a particular biomarker or cell in a biological sample may be considered in reference to a particular threshold level, or in reference to the level of that particular biomarker or cell in a biological sample obtained from the subject prior to commencement of the treatment cycle or obtained from the subject at an earlier timepoint during treatment. In some embodiments, the change in level of the biomarker or cell from the previously measured reference level is determined.

[0107] The level of a particular biomarker or call in a biological sample may indicate that likelihood that the subject will respond to the treatment (i.e., be a responder or a non-responder). Accordingly, in some embodiments, if the level of a particular biomarker or cell is below a pre-determined threshold level, or the change in the level of a particular biomarker or cell relative to a previously measured reference level is below a pre-determined level, then the treatment is discontinued. In some embodiments, if the level of a particular biomarker or cell is above a pre-determined threshold level, or the change in the level of a particular biomarker or cell relative to a previously measured reference level is above a predetermined level, then the treatment is discontinued. In some embodiments, discontinuing treatment includes administering no more doses of either immune checkpoint inhibitor or plinabulin. In some embodiments, discontinuing treatment includes conducting no further treatment cycles that include the administration of both an immune checkpoint inhibitor and plinabulin. In some embodiments, the biological sample may be a tumor biopsy. In other embodiments, the biological sample may be a blood sample. In some such embodiments, the blood sample is a peripheral blood sample. In some embodiments, the peripheral blood sample included peripheral blood mononuclear cells (PMBC).

[0108] In some embodiments, treatment may be discontinued if the level of a particular biomarker in a biological sample is below a particular threshold level. In some embodiments, treatment may be discontinued if the level of a particular biomarker in a biological sample is about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90 percent, or a range between any two of these amounts, below a particular threshold level. Insome embodiments, treatment may be discontinued if the level of a particular biomarker in a biological sample is above a particular threshold level. In some embodiments, treatment may be discontinued if the level of a particular biomarker in a biological sample is about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 percent or more, or a range between any two of these amounts, above a particular threshold level.

[0109] In some embodiments, treatment may be discontinued if the precent decrease in the level of a particular biomarker in a biological sample is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90, or a range between any two of these amounts, below a reference level. In some embodiments, treatment may be discontinued if the precent increase in the level of a particular biomarker in a biological sample is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or more, or a range between any two of these amounts, above a reference level. In some such embodiments, the reference level is the level of the biomarker in a biological sample obtained from a subject prior to commencement of the treatment cycle, e.g., on day 1 of the treatment cycle prior to administration of any therapeutic agents and / or radiation.

[0110] In some embodiments, the reduction in the expression of CCR7 by dendritic cells or a subset thereof in a biological sample of a subject after administration of the one or more checkpoint inhibitor and / or plinabulin and / or radiation therapy as described herein as compared to the expression of CCR7 by dendritic cells or a subset thereof in a biological sample of a subject taken prior to administration of the one or more checkpoint inhibitor and / or plinabulin and / or radiation therapy may determine whether treatment is discontinued. Some patients (Responders) will have a smaller decrease in the expression of CCR7 by dendritic cells or a subset thereof in a biological sample taken on day 4 of the treatment cycle as compared to the level of CCR7 by dendritic cells or a subset thereof in a biological sample taken on day 1 of the treatment cycle prior to administration of the one or more checkpoint inhibitor and plinabulin than other patients (Non-responders). In some embodiments, treatment may be discontinued if the expression of CCR7 by dendritic cells or a subset thereof in a biological sample obtained from a subject on day 4 of the treatment cycle is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90 percent, or a range between any two of these amounts, lower than the expression of CCR7 by dendritic cells or a subset thereof in a biological sample obtained from a subject on day 1 of the treatment cycle prior tothe administration of the one or more checkpoint inhibitor and plinabulin, or prior to day 1 of the treatment cycle. In some embodiments the cells arc plasmacytoid dendritic cells (pDC). In other embodiments, the cells are conventional dendritic cells (cDC).

[0111] In some embodiments, the reduction in the expression of CD80 by dendritic cells or a subset thereof in a biological sample of a subject after administration of the one or more checkpoint inhibitor and / or plinabulin and / or radiation therapy as described herein as compared to the expression of CD80 by dendritic cells or a subset thereof in a biological sample of a subject taken prior to administration of the one or more checkpoint inhibitor and / or plinabulin and / or radiation therapy may determine whether treatment is discontinued. Some patients (Responders) will have a smaller decrease in the expression of CD80 by dendritic cells or a subset thereof in a biological sample taken on day 4 of the treatment cycle as compared to the level of CD80 by dendritic cells or a subset thereof in a biological sample taken on day 1 of the treatment cycle prior to administration of the one or more checkpoint inhibitor and plinabulin than other patients (Non-responders). In some embodiments, treatment may be discontinued if the expression of CD80 by dendritic cells or a subset thereof in a biological sample obtained from a subject on day 4 of the treatment cycle is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90 percent, or a range between any two of these amounts, lower than the expression of CD80 by dendritic cells or a subset thereof in a biological sample obtained from a subject on day 1 of the treatment cycle prior to the administration of the one or more checkpoint inhibitor and plinabulin, or prior to day 1 of the treatment cycle. In some embodiments the cells are plasmacytoid dendritic cells (pDC). In other embodiments, the cells are conventional dendritic cells (cDC).

[0112] In some embodiments, the reduction in the expression of CD83 by dendritic cells or a subset thereof in a biological sample of a subject after administration of the one or more checkpoint inhibitor and / or plinabulin and / or radiation therapy as described herein as compared to the expression of CD83 by dendritic cells or a subset thereof in a biological sample of a subject taken prior to administration of the one or more checkpoint inhibitor and / or plinabulin and / or radiation therapy may determine whether treatment is discontinued. Some patients (Responders) will have a smaller decrease in the expression of CD83 by dendritic cells or a subset thereof in a biological sample taken on day 4 of the treatment cycle as compared to the level of expression of CD83 by dendritic cells or a subset thereof in a biological sampletaken on day 1 of the treatment cycle prior to administration of the one or more checkpoint inhibitor and plinabulin than other patients (Non-rcspondcrs). In some embodiments, treatment may be discontinued if the expression of CD83 by dendritic cells or a subset thereof in a biological sample obtained from a subject on day 4 of the treatment cycle is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90 percent, or a range between any two of these amounts, lower than the expression of CD83 by dendritic cells or a subset thereof in a biological sample obtained from a subject on day 1 of the treatment cycle prior to the administration of the one or more checkpoint inhibitor and plinabulin, or prior to day 1 of the treatment cycle. In some embodiments the cells are plasmacytoid dendritic cells (pDC). In other embodiments, the cells are conventional dendritic cells (cDC).

[0113] In some embodiments, the reduction in the expression of CD86 by dendritic cells or a subset thereof in a biological sample of a subject after administration of the one or more checkpoint inhibitor and / or plinabulin and / or radiation therapy as described herein as compared to the expression of CD86 by dendritic cells or a subset thereof in a biological sample of a subject taken prior to administration of the one or more checkpoint inhibitor and / or plinabulin and / or radiation therapy may determine whether treatment is discontinued. Some patients (Responders) will have a smaller decrease in the expression of CD83 by dendritic cells or a subset thereof in a biological sample taken on day 4 of the treatment cycle as compared to the level of expression of CD83 by dendritic cells or a subset thereof in a biological sample taken on day 1 of the treatment cycle prior to administration of the one or more checkpoint inhibitor and plinabulin than other patients (Non-responders). In some embodiments, treatment may be discontinued if the expression of CD83 by dendritic cells or a subset thereof in a biological sample obtained from a subject on day 4 of the treatment cycle is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90 percent, or a range between any two of these amounts, lower than the expression of CD86 by dendritic cells or a subset thereof in a biological sample obtained from a subject on day 1 of the treatment cycle prior to the administration of the one or more checkpoint inhibitor and plinabulin, or prior to day 1 of the treatment cycle. In some embodiments the cells are plasmacytoid dendritic cells (pDC). In other embodiments, the cells are conventional dendritic cells (cDC).

[0114] In some aspects, provided herein is a method of treating cancer in a subject, the method comprising: (i) obtaining a biological sample from the subject; (ii) determining thelevels of one or more biomarkers or levels of one or more cells in the biological sample; and (iii) administering plinabulin or a pharmaceutically acceptable salt thereof to the subject if the levels of one or more biomarkers or levels of one or more cells in the biological sample are above or below a threshold level. In some embodiments, the method may further include administration of radiation therapy, an immune checkpoint inhibitor, a chemotherapeutic agent, or a combination thereof, to the subject. In some such embodiments, the method may include administration of radiation therapy and an immune checkpoint inhibitor to the subject.

[0115] In some embodiments, the subject may be immuno-oncology refractory. In other embodiments, the subject may be immuno-oncology native.

[0116] In some embodiments, the biological sample is a tumor biopsy. In some embodiments, the biological sample may be a blood sample. In some embodiments, the biological sample is peripheral blood. In some embodiments, the one or more cells are dendritic cells. In some specific embodiments, the dendritic cells are DC3 cells, mDC cells, pDC cells, eDC cells (cDCl and cDC2 cells). In other embodiments, the cells are monocyte- derived macrophage cells (MoMac) cells. In such embodiments, the cells may be MoMac-III cells. In some embodiments, the one or more biomarker is Guanine nucleotide exchange factor- (GEF-H1) immune activation score.

[0117] The GEF-H1 immune pathway includes 47 genes that drives a distinct cell signaling program in DCs. This program is dominated by the c-Jun N-terminal kinase (JNK) pathway and AP-l / ATF transcriptional response for control of innate and adaptive immune responses. In some embodiments, GEF-H1 immune activation score may be determined based the expression levels of one or more of these 47 genes. In one embodiment, GEF-H1 immune activation scores are obtained by first normalizing the gene expression counts by log 2 Fragments Per Kilobase of transcript per Million mapped reads (FPKM) and then summing the expression of these genes to establish a normalized GEF-H1 immune activation score. A discussion of GEF-H1 is provided in Kashyap A. et al., Cell Rep. 2019 September 24; 28(13): 3367-3380, which is incorporated herein by reference in its entirety. The genes in the GEF- H1 immune pathway are listed in the table below.

[0118] In some embodiments, a GEF-H1 immune activation score may be calculated using all 47 genes in the GEF-H1 immune pathway. In other embodiments, a GEF- H1 immune activation score may be calculated using gene expression levels of a subset of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, or 46 genes in the GEF-H1 immune pathway.

[0119] In some embodiments, the GEF-H1 immune activation score can be determined for various cell types, including but not limited to, T-cells (including CD4+ TEM, CD4+ TCM, CD8+ TEM, CD8+TCM, gdT, CD8 naive, dnT, MAIT, CD4 naive, CD4 proliferating, and Treg), NK-cells (including NK, NK proliferating and NK_CD56bright), B- cells (including naive, intermediate, memory, and plamablasts), monocytes (including CD 16+ and CD14+ monocytes), and dendritic cells (including cDCl, cDC2, DC3, mregDC and pDC). In some embodiments, In some embodiments, GEF-H1 immune activation scores can be determined for CD16+ and CD14+ monocytes, NK cells, CD8+ TEM, CD4+ TCM, CD4+ naive T-cells, naive B-cells, cDCl cells and cDC2 cells. In some embodiments, the cells samples may be from a tumor biopsy or from a peripheral blood sample. In some embodiments, the GEF-H1 immune activation score determined for a particular cell type may be higher in responder subjects than in non-responder subjects. Accordingly, in some embodiments, therapy is administered to subjects having a GEF-H1 immune activation score above a pre-determined threshold. For each of the aforementioned cell types, the threshold level of GEF-H1 immune activation score may be about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50, or more, or within a range defined by any two of the aforementioned scores. For example, the threshold level of GEF-H1 score may be fromabout 1 to about 50, from about 10 to about 20, from about 10 to about 30, from about 20 to about 40, or from about 5 to about 30.

[0120] In some embodiments, wherein the cells arc dendritic cells, the threshold level of GEF-H1 immune activation score may be about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50, or more, or within a range defined by any two of the aforementioned scores. For example, the threshold level of GEF-H1 immune activation score may be from about 1 to about 50, from about 10 to about 20, from about 10 to about 30, from about 20 to about 40, or from about 5 to about 30. In some embodiments, wherein the cells are monocyte-derived macrophage cells, the threshold level of GEF-H1 may be about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50. For example the threshold level of GEF-H1 immune activation score may be from about 1 to about 50, from about 10 to about 20, from about 10 to about 30, from about 20 to about 40, or from about 5 to about 30.

[0121] In some embodiments, a level of GEF-H1 score below a threshold level indicates that a patient may not respond to treatment. Accordingly, treatment may not be commenced if GEF-H1 immune activation scores measured from tumor biopsy or a peripheral blood sample are below a threshold level.

[0122] As used herein, the terms “co-administer,” “co-administering,” or “coadministration,” refers to two or more agents or therapies that have a biological effect on a subject at the same time, regardless of when or how they are actually administered. In one embodiment, the agents or therapies are administered simultaneously. In one such embodiment, administration in combination is accomplished by combining the agents in a single dosage form. In another embodiment, the agents or therapies are administered sequentially. In some embodiments, the administration may be separated by a period of time, for example, 30 minutes, 1 hour, 2 hours, 1 day, 2 days, 3 days, or 1 week. In one embodiment the agents are administered through the same route, such as orally. In another embodiment, the agents are administered through different routes, such as one being administered orally and another being administered intravenously.

[0123] In some embodiments, a method for treating cancer in a subject may include co-administcrcd a therapeutically effective amount of plinabulin, or a pharmaceutically acceptable salt thereof, with one or more immune checkpoint inhibitor. In some embodiments, the plinabulin is administered after administration of the one or more immune checkpoint inhibitor. In some embodiments, when the one or more immune checkpoint inhibitor and the plinabulin are administered on the same day, the plinabulin is administered from about 0.5 hours to about 3 hours after completion of administration of the one or more immune checkpoint inhibitor. For example, when the one or more immune checkpoint inhibitor and the plinabulin are administered on the same day, the plinabulin is administered 0.5 hours, 1 hour, 1.5 hours, 2 hours, 2.5 hours, or 3 hours, or a range between any two of these times, after administration of the one or more immune checkpoint inhibitor.

[0124] In some embodiments, the method of treating cancer described herein may include further co-administering an additional chemotherapeutic agent.

[0125] In some embodiments, the method of treating cancer described herein may include co-administering radiation therapy. In some such embodiments, radiation therapy may be administered on day 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, and / or 28 of the treatment cycle. For example, in some embodiments, the radiation therapy may be administered on day 1, 2, and 3 of the treatment cycle. In other embodiments, radiation therapy may be administered on day 1, 2, 3, and 4 of the treatment cycle. In yet other embodiments, radiation therapy may be administered on day 1, 2, 3, 4, and 5 of the treatment cycle. In still yet other embodiments, radiation therapy may be administered on day 2, 3, and 4 of the treatment cycle.

[0126] The total dose of radiation administered to the subject during the treatment cycle can be about 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, or 20 Gy, or more, or within a range defined by any two of the aforementioned values. For example, the total dose of radiation administered to the subject can be from about 1 Gy to about 20 Gy, from about 2 Gy to about 15 Gy, from about 4 Gy to about 15 Gy. In some embodiments, the total dose of radiation administered to the subject is about 4 Gy. In other embodiments, the total dose of radiation administered to the subject is about 8 Gy. the total dose of radiation administered to the subject is about 12.5 Gy. In some embodiments, radiation may beadministered in 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10 fractions, or more. In some embodiments, radiation may be administered in three to five fractions. In some embodiments, radiation may be administered in three fractions. In some embodiments, radiation may be administered in four fractions. In some embodiments, radiation may be administered in five fractions.

[0127] In some embodiments, when radiation therapy and plinabulin are administered on the same day, the plinabulin is administered from about 3 hours to about 12 hours after completion of administration of the radiation therapy. For example, in some such embodiments, plinabulin is administered from about 4 hours to about 10 hours, about 4 hours to about 8 hours, or about 5 hours to about 8 hours after completion of administration of the radiation therapy.

[0128] In some embodiments, the one or more immune checkpoint inhibitor is administered on day 1 of the treatment cycle and plinabulin is administered on day 1 of the treatment cycle. In other embodiments, the one or more immune checkpoint inhibitor is administered on day 1 of the treatment cycle and plinabulin is administered on day 1 and day 4 of the treatment cycle. In some embodiments, the one or more immune checkpoint inhibitor is administered on day 1 of the treatment cycle, plinabulin is administered on day 1 and day 4 of the treatment cycle, and radiation therapy is administered days 1, 2, and 3 of the treatment cycle. In some embodiments, the one or more immune checkpoint inhibitor is administered on day 1 of the treatment cycle, plinabulin is administered on day 1 and day 4 of the treatment cycle, and radiation therapy is administered days 1, 2, 3, and 4 of the treatment cycle. In some embodiments, the one or more immune checkpoint inhibitor is administered on day 1 of the treatment cycle, plinabulin is administered on day 1 and day 4 of the treatment cycle, and radiation therapy is administered days 1, 2, 3, 4, and 5 of the treatment cycle. In some embodiments, the one or more immune checkpoint inhibitor is administered on day 1 and day 15 of the treatment cycle, plinabulin is administered on day 1 and day 4 of the treatment cycle, and radiation therapy is administered days 1, 2, and 3 of the treatment cycle. In some embodiments, the one or more immune checkpoint inhibitor is administered on day 1 and day 15 of the treatment cycle, plinabulin is administered on day 1 and day 4 of the treatment cycle, and radiation therapy is administered days 1, 2, 3, and 4 of the treatment cycle. In some embodiments, the one or more immune checkpoint inhibitor is administered on day 1 and day15 of the treatment cycle, plinabulin is administered on day 1 and day 4 of the treatment cycle, and radiation therapy is administered days 1, 2, 3, 4, and 5 of the treatment cycle.

[0129] In some embodiments, radiation therapy may be administered during each treatment cycle. In other embodiments, radiation therapy is administered during some treatment cycles. For example, in some embodiments, radiation therapy is administered only during the first treatment cycle. In other embodiments, radiation therapy is administered during the first and second treatment cycles. In other embodiments, radiation therapy is administered during the first, second, and third treatment cycles.

[0130] In some embodiments, plinabulin is administered on day 1 and day 4 of the first treatment cycle and on day 1 of each subsequent (i.e., second, third, fourth, fifth) treatment cycle. In some embodiments, plinabulin is administered on day 1 and day 4 of the first and second treatment cycle and on day 1 of each subsequent treatment cycle.

[0131] In some embodiments, the one or more checkpoint inhibitor is administered on day 1 of the first treatment cycle and on day 1 of each subsequent (i.e., second, third, fourth, fifth) treatment cycle. In some embodiments, one or more checkpoint inhibitor is administered on day 1 and day 15 of the first treatment cycle and on day 1 of each subsequent treatment cycle. In some embodiments, one or more checkpoint inhibitor is administered on day 1 and day 15 of the first treatment cycle and on day 1 and 15 of each subsequent treatment cycle.

[0132] In some embodiments, the present disclosure provides a method for treating a breast cancer, a bladder cancer, a glioma, a glioblastoma, a head and neck cancer, a nonsmall cell lung cancer, a small cell lung cancer, recurrent small cell lung cancer (SCLC), a colorectal cancer, a gastrointestinal stromal tumor, a gastroesophageal carcinoma, a renal cell cancer, a prostate cancer, a liver cancer, a colon cancer, a pancreatic cancer, an ovarian cancer, a lymphoma, or a cutaneous T-cell lymphoma, or a melanoma.

[0133] In some embodiments, the present disclosure provides a method for treating Fibrolamellar hepatocellular carcinoma, MSI-H cancers of any histology including but not limited to: colorectal, endometrial, adrenocortical, anal, appendiceal cancer, biliary, bladder, brain, breast, cervical, gastric or gastroesophageal junction, head and neck squamous cell, liver, mesothelioma, nasopharyngeal, neuroendocrine, ovarian, pancreatic, prostate, renal cell, retroperitoneal, salivary, sarcoma, small cell lung, small intestinal, testicular, thyroid, vaginaland vulvar. In some embodiments, the present disclosure provides a method for treating tumor type that has a checkpoint inhibitor approval.

[0134] In some embodiments, the subject can be an animal, e.g., a mammal, a human. In some embodiments, the subject is a human.

[0135] In some embodiments, plinabulin or a pharmaceutically acceptable salt thereof is incorporated in a pharmaceutically acceptable solution. In some embodiments, plinabulin or a pharmaceutically acceptable salt thereof is incorporated in an injectable formulation. In some embodiments, plinabulin or a pharmaceutically acceptable salt thereof is incorporated in an injectable formulation that substantially maintains plinabulin or a pharmaceutically acceptable salt thereof at or near the injection site.

[0136] The precise amount of plinabulin or a pharmaceutically acceptable salt thereof incorporated in a particular method or therapeutic combination of the disclosure may vary according to factors known in art such as for example, the physical and clinical status of the subject, the method of administration, the content of the formulation, the intended dosing regimen or sequence. Accordingly, it is not practical to specifically set forth an amount that constitutes an amount of plinabulin or a pharmaceutically acceptable salt thereof therapeutically effective for all possible applications. Those of ordinary skill in the art, however, can readily determine an appropriate amount with due consideration of such factors.Administration

[0137] Administration of the pharmaceutical compositions described herein can be via any of the accepted modes of administration for agents that serve similar utilities including, but not limited to, orally, sublingually, buccally, subcutaneously, intravenously, intranasally, intratumor ally, topically, transdermally, intradermally, intraperitoneally, intramuscularly, intrapulmonarilly, vaginally, rectally, or intraocularly. Oral and parenteral administrations are customary in treating the indications that are the subject of the preferred embodiments.

[0138] The compositions described herein may be provided in unit dosage form. As used herein, a "unit dosage form" is a composition containing an amount of a compound or composition that is suitable for administration to an animal, preferably mammal subject, in a single dose, according to good medical practice. The preparation of a single or unit dosage form however, does not imply that the dosage form is administered once per day or once percourse of therapy. Such dosage forms are contemplated to be administered once, twice, thrice or more per day and may be administered as infusion over a period of time (c.g., from about 30 minutes to about 2-6 hours), or administered as a continuous infusion, and may be given more than once during a course of therapy, although a single administration is not specifically excluded. The skilled artisan will recognize that the formulation does not specifically contemplate the entire course of therapy and such decisions are left for those skilled in the art of treatment rather than formulation.

[0139] The compositions useful as described above may be in any of a variety of suitable forms for a variety of routes for administration, for example, for oral, sublingual, buccal, nasal, rectal, topical (including transdermal and intradermal), ocular, intracerebral, intracranial, intrathecal, intra-arterial, intravenous, intramuscular, or other parental routes of administration. The skilled artisan will appreciate that oral and nasal compositions include compositions that are administered by inhalation, and made using available methodologies. Depending upon the particular route of administration desired, a variety of pharmaceutically- acceptable carriers well-known in the art may be used. Pharmaceutically-acceptable carriers include, for example, solid or liquid fillers, diluents, hydrotropies, surface-active agents, and encapsulating substances. Optional pharmaceutically-active materials may be included, which do not substantially interfere with the inhibitory activity of the compound or composition. The amount of carrier employed in conjunction with the compound or composition is sufficient to provide a practical quantity of material for administration per unit dose of the compound. Techniques and compositions for making dosage forms useful in the methods described herein are described in the following references, all incorporated by reference herein: Modem Pharmaceutics, 4th Ed., Chapters 9 and 10 (Banker & Rhodes, editors, 2002); Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1989); and Ansel, Introduction to Pharmaceutical Dosage Forms 8th Edition (2004).

[0140] Various oral dosage forms can be used, including such solid forms as tablets, capsules (e.g. solid gel capsules and liquid gel capsules), granules and bulk powders. Tablets can be compressed, tablet triturates, enteric-coated, sugar-coated, film-coated, or multiple-compressed, containing suitable binders, lubricants, diluents, disintegrating agents, coloring agents, flavoring agents, flow-inducing agents, and melting agents. Liquid oral dosage forms include aqueous solutions, emulsions, suspensions, solutions and / or suspensionsreconstituted from non-effervescent granules, and effervescent preparations reconstituted from effervescent granules, containing suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, melting agents, coloring agents and flavoring agents.

[0141] The pharmaceutically-acceptable carriers suitable for the preparation of unit dosage forms for peroral administration is well-known in the art. Tablets typically comprise conventional pharmaceutically-compatible adjuvants as inert diluents, such as calcium carbonate, sodium carbonate, mannitol, lactose and cellulose; binders such as starch, gelatin and sucrose; disintegrants such as starch, alginic acid and croscarmelose; lubricants such as magnesium stearate, stearic acid and talc. Glidants such as silicon dioxide can be used to improve flow characteristics of the powder mixture. Coloring agents, such as the FD&C dyes, can be added for appearance. Sweeteners and flavoring agents, such as aspartame, saccharin, menthol, peppermint, and fruit flavors, are useful adjuvants for chewable tablets. Capsules typically comprise one or more solid diluents disclosed above. The selection of carrier components depends on secondary considerations like taste, cost, and shelf stability, which are not critical, and can be readily made by a person skilled in the art.

[0142] Peroral compositions also include liquid solutions, emulsions, suspensions, and the like. The pharmaceutically-acceptable carriers suitable for preparation of such compositions are well known in the art. Typical components of carriers for syrups, elixirs, emulsions and suspensions include ethanol, glycerol, propylene glycol, polyethylene glycol, liquid sucrose, sorbitol and water. For a suspension, typical suspending agents include methyl cellulose, sodium carboxymethyl cellulose, AVICEL RC-591, tragacanth and sodium alginate; typical wetting agents include lecithin and polysorbate 80; and typical preservatives include methyl paraben and sodium benzoate. Peroral liquid compositions may also contain one or more components such as sweeteners, flavoring agents and colorants disclosed above.

[0143] Such compositions may also be coated by conventional methods, typically with pH or time-dependent coatings, such that the subject composition is released in the gastrointestinal tract in the vicinity of the desired topical application, or at various times to extend the desired action. Such dosage forms typically include, but are not limited to, one or more of cellulose acetate phthalate, polyvinylacetate phthalate, hydroxypropyl methyl cellulose phthalate, ethyl cellulose, Eudragit coatings, waxes and shellac.

[0144] Compositions described herein may optionally include additional drug actives.

[0145] Other compositions useful for attaining systemic delivery of the subject compounds include sublingual, buccal and nasal dosage forms. Such compositions typically comprise one or more of soluble filler substances such as sucrose, sorbitol and mannitol; and binders such as acacia, microcrystalline cellulose, carboxymethyl cellulose and hydroxypropyl methyl cellulose. Glidants, lubricants, sweeteners, colorants, antioxidants and flavoring agents disclosed above may also be included.

[0146] A liquid composition, which is formulated for topical ophthalmic use, is formulated such that it can be administered topically to the eye. The comfort may be maximized as much as possible, although sometimes formulation considerations (e.g. drug stability) may necessitate less than optimal comfort. In the case that comfort cannot be maximized, the liquid may be formulated such that the liquid is tolerable to the patient for topical ophthalmic use. Additionally, an ophthalmically acceptable liquid may either be packaged for single use, or contain a preservative to prevent contamination over multiple uses.

[0147] For ophthalmic application, solutions or medicaments are often prepared using a physiological saline solution as a major vehicle. Ophthalmic solutions may preferably be maintained at a comfortable pH with an appropriate buffer system. The formulations may also contain conventional, pharmaceutically acceptable preservatives, stabilizers and surfactants.

[0148] Preservatives that may be used in the pharmaceutical compositions disclosed herein include, but are not limited to, benzalkonium chloride, PHMB, chlorobutanol, thimerosal, phenylmercuric, acetate and phenylmercuric nitrate. A useful surfactant is, for example, Tween 80. Likewise, various useful vehicles may be used in the ophthalmic preparations disclosed herein. These vehicles include, but are not limited to, polyvinyl alcohol, povidone, hydroxypropyl methyl cellulose, poloxamers, carboxymethyl cellulose, hydroxyethyl cellulose and purified water.

[0149] Tonicity adjustors may be added as needed or convenient. They include, but are not limited to, salts, particularly sodium chloride, potassium chloride, mannitol and glycerin, or any other suitable ophthalmically acceptable tonicity adjustor.

[0150] Various buffers and means for adjusting pH may be used so long as the resulting preparation is ophthalmically acceptable. For many compositions, the pH will be between 4 and 9. Accordingly, buffers include acetate buffers, citrate buffers, phosphate buffers and borate buffers. Acids or bases may be used to adjust the pH of these formulations as needed.

[0151] Ophthalmically acceptable antioxidants include, but are not limited to, sodium metabisulfite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole and butylated hydroxytoluene.

[0152] Other excipient components, which may be included in the ophthalmic preparations, are chelating agents. A useful chelating agent is edetate disodium, although other chelating agents may also be used in place or in conjunction with it.

[0153] For topical use, creams, ointments, gels, solutions or suspensions, etc., containing the composition disclosed herein are employed. Topical formulations may generally be comprised of a pharmaceutical carrier, co-solvent, emulsifier, penetration enhancer, preservative system, and emollient.

[0154] For intravenous administration, the compositions described herein may be dissolved or dispersed in a pharmaceutically acceptable diluent, such as a saline or dextrose solution. Suitable excipients may be included to achieve the desired pH, including but not limited to NaOH, sodium carbonate, sodium acetate, HO, and citric acid. In various embodiments, the pH of the final composition ranges from 2 to 8, or preferably from 4 to 7. Antioxidant excipients may include sodium bisulfite, acetone sodium bisulfite, sodium formaldehyde, sulfoxylate, thiourea, and EDTA. In some embodiments, excipients utilized for intravenous delivery may include Kolliphor HS 15 (polyoxyl 15 hydroxystearate or Solutol HS-15), propylene glycol and 5% dextrose in water (D5W). Other non-limiting examples of suitable excipients found in the final intravenous composition may include sodium or potassium phosphates, citric acid, tartaric acid, gelatin, and carbohydrates such as dextrose, mannitol, and dextran. Further acceptable excipients are described in Powell, et al., Compendium of Excipients for Parenteral Formulations, PDA J Pharm Sci and Tech 1998, 52 238-311 and Nema et al., Excipients and Their Role in Approved Injectable Products: Current Usage and Future Directions, PDA J Pharm Sci and Tech 2011, 65287-332, both of which are incorporated herein by reference in their entirety. Antimicrobial agents may also be includedto achieve a bacteriostatic or fungistatic solution, including but not limited to phenylmercuric nitrate, thimcrosal, benzethonium chloride, benzalkonium chloride, phenol, cresol, and chlorobutanol.

[0155] The compositions for intravenous administration may be provided to caregivers in the form of one more solids that are reconstituted with a suitable diluent such as sterile water, saline or dextrose in water shortly prior to administration. In other embodiments, the compositions are provided in solution ready to administer parenterally. In still other embodiments, the compositions are provided in a solution that is further diluted prior to administration. In embodiments that include administering a combination of a compound described herein and another agent, the combination may be provided to caregivers as a mixture, or the caregivers may mix the two agents prior to administration, or the two agents may be administered separately.

[0156] The actual dose of the active compounds described herein depends on the specific compound, and on the condition to be treated; the selection of the appropriate dose is well within the knowledge of the skilled artisan. In some embodiments, plinabulin may be administered at a dose in the range of about 1 mg / m2to about 50 mg / m2. In some embodiments, plinabulin is administered at a dose in the range of about 1-50 mg / m2of the body surface area. In some embodiments, plinabulin is administered at a dose in the range of about 1-2, 1-3, 1- 4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 1-11, 1-12, 1-13, 1-13.75, 1-14, 1-15, 1-16, 1-17, 1-18, 1-19, 1-20, 1-22.5, 1-25, 1-27.5, 1-30, 1.5-2, 1.5-3, 1.5-4, 1.5-5, 1.5-6, 1.5-7, 1.5-8, 1.5-9, 1.5-10,1.5-11, 1.5-12, 1.5-13, 1.5-13.75, 1.5-14, 1.5-15, 1.5-16, 1.5-17, 1.5-18, 1.5-19, 1.5-20, 1.5- 22.5, 1.5-25, 1.5-27.5, 1.5-30, 2.5-2, 2.5-3, 2.5-4, 2.5-5, 2.5-6, 2.5-7, 2.5-8, 2.5-9, 2.5-10, 2.5- 11, 2.5-12, 2.5-13, 2.5-13.75, 2.5-14, 2.5-15, 2.5-16, 2.5-17, 2.5-18, 2.5-19, 2.5-20, 2.5-22.5,2.5-25, 2.5-27.5, 2.5-30, 2.5-7.5, 3-4, 3-5, 3-6, 3-7, 3-8, 3-9, 3-10, 3-11, 3-12, 3-13, 3-13.75,3-14, 3-15, 3-16, 3-17, 3-18, 3-19, 3-20, 3-22.5, 3-25, 3-27.5, 3-30, 3.5- 6.5, 3.5-13.75, 3.5-15,2.5-17.5, 4-5, 4-6, 4-7, 4-8, 4-9, 4-10, 4-11, 4-12, 4-13, 4-13.75, 4-14, 4-15, 4-16, 4-17, 4-18,4-19, 4-20, 4-22.5, 4-25, 4-27.5, 4-30, 5-6, 5-7, 5-8, 5-9, 5-10, 5-11, 5-12, 5-13, 5-13.75, 5-14,5-15, 5-16, 5-17, 5-18, 5-19, 5-20, 5-22.5, 5-25, 5-27.5, 5-30, 6-7, 6-8, 6-9, 6-10, 6-11, 6-12,6-13, 6-13.75, 6-14, 6-15, 6-16, 6-17, 6-18, 6-19, 6-20, 6-22.5, 6-25, 6-27.5, 6-30, 7-8, 7-9, 7- 10, 7-11, 7-12, 7-13, 7-13.75, 7-14, 7-15, 7-16, 7-17, 7-18, 7-19, 7-20, 7-22.5, 7-25, 7-27.5, 7- 30, 7.5-12.5, 7.5-13.5, 7.5-15, 8-9, 8-10, 8-11, 8-12, 8-13, 8-13.75, 8-14, 8-15, 8-16, 8-17, 8-18, 8-19, 8-20, 8-22.5, 8-25, 8-27.5, 8-30, 9-10, 9-1 1 , 9-12, 9-13, 9- 13.75, 9-14, 9- 15, 9-16, 9- 17, 9-18, 9-19, 9-20, 9-22.5, 9-25, 9-27.5, 9-30, 10-11, 10-12, 10-13, 10-13.75, 10-14, 10-15, 10-16, 10-17, 10-18, 10-19, 10-20, 10-22.5, 10-25, 10-27.5, 10-30, 11.5-15.5, 12.5-14.5, 7.5-22.5, 8.5-32.5, 9.5-15.5, 15.5-24.5, 5-35, 17.5-22.5, 22.5-32.5, 25-35, 25.5-34.5, 27.5-32.5, 2- 20, 2.5-22.5, or 9.5-21.5 mg / m2, of the body surface area. In some embodiments, plinabulin is administered at a dose of about 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9,9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19,19.5, 20, 20.5, 21, 21.5, 22, 22.5, 23, 23.5, 24, 24.5, 25, 25.5, 26, 26.5, 27, 27.5, 28, 28.5, 29,29.5, 30, 30.5, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40 mg / m2of the body surface area. In some embodiments, plinabulin is administered at a dose less than about 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4,4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, 20, 20.5, 21, 21.5, 22, 22.5, 23, 23.5, 24, 24.5, 25, 25.5, 26, 26.5, 27, 27.5, 28, 28.5, 29, 29.5, 30, 30.5, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40 mg / m2of the body surface area. In some embodiments, plinabulin is administered at a dose greater than about 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12,12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, 20, 20.5, 21, 21.5, 22,22.5, 23, 23.5, 24, 24.5, 25, 25.5, 26, 26.5, 27, 27.5, 28, 28.5, 29, 29.5, 30, 30.5, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 mg / m2of the body surface area.

[0157] In some embodiments, the plinabulin dose is about 5 mg - 300 mg, 5 mg - 200 mg, 7.5 mg - 200 mg, 10 mg - 100 mg, 15 mg - 100 mg, 20 mg - 100 mg, 30 mg - 100 mg, 40 mg - 100 mg, 10 mg - 80 mg, 15 mg - 80 mg, 20 mg - 80 mg, 30 mg - 80 mg, 40 mg - 80 mg, 10 mg - 60 mg, 15 mg - 60 mg, 20 mg - 60 mg, 30 mg - 60 mg, or about 40 mg - 60 mg. In some embodiments, the dose of plinabulin administered is about 20 mg - 60 mg, 27 mg - 60 mg, 20 mg - 45 mg, or 27 mg - 45 mg. In some embodiments, the dose of plinabulin administered is about 5 mg-7.5 mg, 5 mg-9 mg, 5 mg- 10 mg, 5 mg-12mg, 5mg-14mg, 5mg- 15 mg, 5 mg- 16 mg, 5 mg- 18 mg, 5 mg-20 mg, 5 mg-22 mg, 5 mg-24 mg, 5 mg-26 mg, 5 mg- 28mg, 5mg-30mg, 5mg-32mg, 5mg-34mg, 5mg-36mg, 5mg-38mg, 5mg-40mg, 5mg-42mg, 5mg-44mg, 5mg-46mg, 5mg-48mg, 5mg-50mg, 5mg-52mg, 5mg-54mg, 5mg-56mg, 5mg- 58mg, 5mg-60mg, 7 mg-7.7 mg, 7 mg-9 mg, 7 mg-10 mg, 7 mg-12mg, 7mg-14mg, 7mg-15 mg, 7 mg- 16 mg, 7 mg- 18 mg, 7 mg-20 mg, 7 mg-22 mg, 7 mg-24 mg, 7 mg-26 mg, 7 mg- 28mg, 7mg-30mg, 7mg-32mg, 7mg-34mg, 7mg-36mg, 7mg-38mg, 7mg-40mg, 7mg-42mg,7mg-44mg, 7mg-46mg, 7mg-48mg, 7mg-50mg, 7mg-52mg, 7mg-54mg, 7mg-56mg, 7mg- 58mg, 7mg-60mg, 9 mg-10 mg, 9 mg-12mg, 9mg-14mg, 9mg-15 mg, 9 mg-16 mg, 9 mg-18 mg, 9 mg-20 mg, 9 mg-22 mg, 9 mg-24 mg, 9 mg-26 mg, 9 mg-28mg, 9mg-30mg, 9mg-32mg, 9mg-34mg, 9mg-36mg, 9mg-38mg, 9mg-40mg, 9mg-42mg, 9mg-44mg, 9mg-46mg, 9mg- 48mg, 9mg-50mg, 9mg-52mg, 9mg-54mg, 9mg-56mg, 9mg-58mg, 9mg-60mg, 10 mg-12mg, 10mg-14mg, 10mg-15 mg, 10 mg-16 mg, 10 mg-18 mg, 10 mg-20 mg, 10 mg-22 mg, 10 mg- 24 mg, 10 mg-26 mg, 10 mg-28mg, 10mg-30mg, 10mg-32mg, 10mg-34mg, 10mg-36mg, 10mg-38mg, 10mg-40mg, 10mg-42mg, 10mg-44mg, 10mg-46mg, 10mg-48mg, 10mg-50mg, 10mg-52mg, 10mg-54mg, 10mg-56mg, 10mg-58mg, 10mg-60mg, 12mg-14mg, 12mg-15 mg, 12 mg-16 mg, 12 mg-18 mg, 12 mg-20 mg, 12 mg-22 mg, 12 mg-24 mg, 12 mg-26 mg, 12 mg-28mg, 12mg-30mg, 12mg-32mg, 12mg-34mg, 12mg-36mg, 12mg-38mg, 12mg-40mg, 12mg-42mg, 12mg-44mg, 12mg-46mg, 12mg-48mg, 12mg-50mg, 12mg-52mg, 12mg-54mg, 12mg-56mg, 12mg-58mg, 12mg-60mg, 15 mg-16 mg, 15 mg-18 mg, 15 mg-20 mg, 15 mg-22 mg, 15 mg-24 mg, 15 mg-26 mg, 15 mg-28mg, 15mg-30mg, 15mg-32mg, 15mg-34mg, 15mg- 36mg, 15mg-38mg, 15mg-40mg, 15mg-42mg, 15mg-44mg, 15mg-46mg, 15mg-48mg, 15mg- 50mg, 15mg-52mg, 15mg-54mg, 15mg-56mg, 15mg-58mg, 15mg-60mg, 17 mg-18 mg, 17 mg-20 mg, 17 mg-22 mg, 17 mg-24 mg, 17 mg-26 mg, 17 mg-28mg, 17mg-30mg, 17mg- 32mg, 17mg-34mg, 17mg-36mg, 17mg-38mg, 17mg-40mg, 17mg-42mg, 17mg-44mg, 17mg- 46mg, 17mg-48mg, 17mg-50mg, 17mg-52mg, 17mg-54mg, 17mg-56mg, 17mg-58mg, 17mg- 60mg, 20 mg-22 mg, 20 mg-24 mg, 20 mg-26 mg, 20 mg-28mg, 20mg-30mg, 20mg-32mg, 20mg-34mg, 20mg-36mg, 20mg-38mg, 20mg-40mg, 20mg-42mg, 20mg-44mg, 20mg-46mg, 20mg-48mg, 20mg-50mg, 20mg-52mg, 20mg-54mg, 20mg-56mg, 20mg-58mg, 20mg-60mg, 22 mg-24 mg, 22 mg-26 mg, 22 mg-28mg, 22mg-30mg, 22mg-32mg, 22mg-34mg, 22mg- 36mg, 22mg-38mg, 22mg-40mg, 22mg-42mg, 22mg-44mg, 22mg-46mg, 22mg-48mg, 22mg- 50mg, 22mg-52mg, 22mg-54mg, 22mg-56mg, 22mg-58mg, 22mg-60mg, 25 mg-26 mg, 25 mg-28mg, 25mg-30mg, 25mg-32mg, 25mg-34mg, 25mg-36mg, 25mg-38mg, 25mg-40mg, 25mg-42mg, 25mg-44mg, 25mg-46mg, 25mg-48mg, 25mg-50mg, 25mg-52mg, 25mg-54mg, 25mg-56mg, 25mg-58mg, 25mg-60mg, 27 mg-28mg, 27mg-30mg, 27mg-32mg, 27mg-34mg, 27mg-36mg, 27mg-38mg, 27mg-40mg, 27mg-42mg, 27mg-44mg, 27mg-46mg, 27mg-48mg, 27mg-50mg, 27mg-52mg, 27mg-54mg, 27mg-56mg, 27mg-58mg, 27mg-60mg, 30mg-32mg, 30mg-34mg, 30mg-36mg, 30mg-38mg, 30mg-40mg, 30mg-42mg, 30mg-44mg, 30mg-46mg,30mg-48mg, 30mg-50mg, 30mg-52mg, 30mg-54mg, 30mg-56mg, 30mg-58mg, 30mg-60mg, 33mg-34mg, 33mg-36mg, 33mg-38mg, 33mg-40mg, 33mg-42mg, 33mg-44mg, 33mg-46mg, 33mg-48mg, 33mg-50mg, 33mg-52mg, 33mg-54mg, 33mg-56mg, 33mg-58mg, 33mg-60mg, 36mg-38mg, 36mg-40mg, 36mg-42mg, 36mg-44mg, 36mg-46mg, 36mg-48mg, 36mg-50mg, 36mg-52mg, 36mg-54mg, 36mg-56mg, 36mg-58mg, 36mg-60mg, 40mg-42mg, 40mg-44mg, 40mg-46mg, 40mg-48mg, 40mg-50mg, 40mg-52mg, 40mg-54mg, 40mg-56mg, 40mg-58mg, 40mg-60mg, 43mg-46mg, 43mg-48mg, 43mg-50mg, 43mg-52mg, 43mg-54mg, 43mg-56mg, 43mg-58mg, 42mg-60mg, 45mg-48mg, 45mg-50mg, 45mg-52mg, 45mg-54mg, 45mg-56mg, 45mg-58mg, 45mg-60mg, 48mg-50mg, 48mg-52mg, 48mg-54mg, 48mg-56mg, 48mg-58mg, 48mg-60mg, 50mg-52mg, 50mg-54mg, 50mg-56mg, 50mg-58mg, 50mg-60mg, 52mg-54mg, 52mg-56mg, 52mg-58mg, or 52mg-60mg. In some embodiments, the plinabulin dose is greater than about 5 mg, about 10 mg, about 12.5 mg, about 13.5 mg, about 15 mg, about 17.5 mg, about 20 mg, about 22.5 mg, about 25 mg, about 27 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, about 150mg, or about 200 mg. In some embodiments, the plinabulin dose is about less than about 5 mg, about 10 mg, about 12.5 mg, about 13.5 mg, about 15 mg, about 17.5 mg, about 20 mg, about 22.5 mg, about 25 mg, about 27 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, about 150mg, or about 200 mg.

[0158] In some embodiments, a dose of one or more immune checkpoint inhibitors may be from about 100 pg to about 1000 mg, from about 500 pg or less to about 800 mg, from about 1.0 mg to about 600 mg, from about 100 mg to about 600 mg, or from about 200 mg to 500 mg. In some embodiments, a dose of one or more immune checkpoint inhibitors may be from about 240 mg to about 480 mg per dose. In some embodiments, the dose of the one or more immune checkpoint inhibitors is about 240 mg. In some embodiments, the dose of the one or more immune checkpoint inhibitors is about 480 mg.

[0159] In some embodiments, one or more immune checkpoint inhibitors may be administered at a dose in the range of about 100 mg / kg to about 5000 mg / kg. In some embodiments, one or more immune checkpoint inhibitors is administered at a dose in the range of about 100-1000 mg / kg. In some embodiments, one or more immune checkpoint inhibitors is administered at a dose in the range of about 100-200, 100-300, 100-400, 100-500, 100-600,100-700, 100-800, 100-900, 100-1000, 100-1 100, 100-1200, 100-1300, 100-1375, 100-1400, 100-1500, 100-1600, 100-1700, 100-1800, 100-1900, 100-2000, 100-2250, 100-2500, 100- 2750, 100-3000, 150-200, 150-300, 150-400, 150-500, 150-600, 150-700, 150-800, 150-900, 150-1000, 150-1100, 150-1200, 150-1300, 150-1375, 150-1400, 150-1500, 150-1600, 150- 1700, 150-1800, 150-1900, 150-2000, 150-2250, 150-2500, 150-2750, 150-3000, 250-2000, 250-3000, 250-4000, 250-5000, 250-600, 250-700, 250-800, 250-900, 250-1000, 250-1100, 250-1200, 250-1300, 250-1375, 250-1400, 250-1500, 250-1600, 250-1700, 250-1800, 250- 1900, 250-2000, 250-2250, 250-2500, 250-2750, 250-3000, 250-750, 300-400, 300-500, 300- 600, 300-700, 300-800, 300-900, 300-1000, 300-1100, 300-1200, 300-1300, 300-1375, 300- 1400, 300-1500, 300-1600, 300-1700, 300-1800, 300-1900, 300-2000, 300-2250, 300-2500, 300-2750, or 300-3000, mg / kg. In some embodiments, one or more immune checkpoint inhibitors is administered at a dose of about 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, 20, 20.5, 21, 21.5, 22, 22.5, 23, 23.5, 24, 24.5, 25, 25.5, 26, 26.5, 27, 27.5, 28, 28.5, 29, 29.5, 30, 30.5, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 mg.

[0160] In some embodiments, one or more immune checkpoint inhibitor dose is about 0.5 mg - 3000 mg, 0.5 mg - 2500 mg, 0.5 mg - 2000 mg, 0.5 mg - 1500 mg, 0.5 mg - 1000 mg, 0.5 mg - 500 mg, 0.5 mg -200 mg, 0.75 mg - 200 mg, 1.0 mg - 100 mg, 1.5 mg - 100 mg, 2.0 mg - 100 mg, 3.0 mg - 100 mg, 4.0 mg - 100 mg, 1.0 mg - 80 mg, 1.5 mg - 80 mg, 2.0 mg - 80 mg, 3.0 mg - 80 mg, 4.0 mg - 80 mg, 1.0 mg - 60 mg, 1.5 mg - 60 mg, 2.0 mg - 60 mg, 3.0 mg - 60 mg, or about 4.0 mg - 60 mg. In some embodiments, one or more immune checkpoint inhibitors administered is about 20 mg - 60 mg, 27 mg - 60 mg, 20 mg - 45 mg, or 27 mg - 45 mg. In some embodiments, one or more immune checkpoint inhibitors administered is about 5 mg-7.5 mg, 5 mg-9 mg, 5 mg-10 mg, 5 mg-12mg, 5mg-14mg, 5mg-15 mg, 5 mg-16 mg, 5 mg- 18 mg, 5 mg-20 mg, 5 mg-22 mg, 5 mg-24 mg, 5 mg-26 mg, 5 mg-28mg, 5mg- 30mg, 5mg-32mg, 5mg-34mg, 5mg-36mg, 5mg-38mg, 5mg-40mg, 5mg-42mg, 5mg-44mg, 5mg-46mg, 5mg-48mg, 5mg-50mg, 5mg-52mg, 5mg-54mg, 5mg-56mg, 5mg-58mg, 5mg- 60mg, 7 mg-7.7 mg, 7 mg-9 mg, 7 mg-10 mg, 7 mg-12mg, 7mg-14mg, 7mg-15 mg, 7 mg-16 mg, 7 mg- 18 mg, 7 mg-20 mg, 7 mg-22 mg, 7 mg-24 mg, 7 mg-26 mg, 7 mg-28mg, 7mg- 30mg, 7mg-32mg, 7mg-34mg, 7mg-36mg, 7mg-38mg, 7mg-40mg, 7mg-42mg, 7mg-44mg,7mg-46mg, 7mg-48mg, 7mg-50mg, 7mg-52mg, 7mg-54mg, 7mg-56mg, 7mg-58mg, 7mg- 60mg, 9 mg-10 mg, 9 mg-12mg, 9mg-14mg, 9mg-15 mg, 9 mg-16 mg, 9 mg-18 mg, 9 mg-20 mg, 9 mg-22 mg, 9 mg-24 mg, 9 mg-26 mg, 9 mg-28mg, 9mg-30mg, 9mg-32mg, 9mg-34mg, 9mg-36mg, 9mg-38mg, 9mg-40mg, 9mg-42mg, 9mg-44mg, 9mg-46mg, 9mg-48mg, 9mg- 50mg, 9mg-52mg, 9mg-54mg, 9mg-56mg, 9mg-58mg, 9mg-60mg, 10 mg-12mg, 10mg-14mg, 10mg-15 mg, 10 mg-16 mg, 10 mg-18 mg, 10 mg-20 mg, 10 mg-22 mg, 10 mg-24 mg, 10 mg- 26 mg, 10 mg-28mg, 10mg-30mg, 10mg-32mg, 10mg-34mg, 10mg-36mg, 10mg-38mg, 10mg-40mg, 10mg-42mg, 10mg-44mg, 10mg-46mg, 10mg-48mg, 10mg-50mg, 10mg-52mg, 10mg-54mg, 10mg-56mg, 10mg-58mg, 10mg-60mg, 12mg-14mg, 12mg-15 mg, 12 mg-16 mg, 12 mg-18 mg, 12 mg-20 mg, 12 mg-22 mg, 12 mg-24 mg, 12 mg-26 mg, 12 mg-28mg, 12mg-30mg, 12mg-32mg, 12mg-34mg, 12mg-36mg, 12mg-38mg, 12mg-40mg, 12mg-42mg, 12mg-44mg, 12mg-46mg, 12mg-48mg, 12mg-50mg, 12mg-52mg, 12mg-54mg, 12mg-56mg, 12mg-58mg, 12mg-60mg, 15 mg-16 mg, 15 mg-18 mg, 15 mg-20 mg, 15 mg-22 mg, 15 mg- 24 mg, 15 mg-26 mg, 15 mg-28mg, 15mg-30mg, 15mg-32mg, 15mg-34mg, 15mg-36mg, 15mg-38mg, 15mg-40mg, 15mg-42mg, 15mg-44mg, 15mg-46mg, 15mg-48mg, 15mg-50mg, 15mg-52mg, 15mg-54mg, 15mg-56mg, 15mg-58mg, 15mg-60mg, 17 mg-18 mg, 17 mg-20 mg, 17 mg-22 mg, 17 mg-24 mg, 17 mg-26 mg, 17 mg-28mg, 17mg-30mg, 17mg-32mg, 17mg-34mg, 17mg-36mg, 17mg-38mg, 17mg-40mg, 17mg-42mg, 17mg-44mg, 17mg-46mg, 17mg-48mg, 17mg-50mg, 17mg-52mg, 17mg-54mg, 17mg-56mg, 17mg-58mg, 17mg-60mg, 20 mg-22 mg, 20 mg-24 mg, 20 mg-26 mg, 20 mg-28mg, 20mg-30mg, 20mg-32mg, 20mg- 34mg, 20mg-36mg, 20mg-38mg, 20mg-40mg, 20mg-42mg, 20mg-44mg, 20mg-46mg, 20mg- 48mg, 20mg-50mg, 20mg-52mg, 20mg-54mg, 20mg-56mg, 20mg-58mg, 20mg-60mg, 22 mg- 24 mg, 22 mg-26 mg, 22 mg-28mg, 22mg-30mg, 22mg-32mg, 22mg-34mg, 22mg-36mg, 22mg-38mg, 22mg-40mg, 22mg-42mg, 22mg-44mg, 22mg-46mg, 22mg-48mg, 22mg-50mg, 22mg-52mg, 22mg-54mg, 22mg-56mg, 22mg-58mg, 22mg-60mg, 25 mg-26 mg, 25 mg- 28mg, 25mg-30mg, 25mg-32mg, 25mg-34mg, 25mg-36mg, 25mg-38mg, 25mg-40mg, 25mg- 42mg, 25mg-44mg, 25mg-46mg, 25mg-48mg, 25mg-50mg, 25mg-52mg, 25mg-54mg, 25mg- 56mg, 25mg-58mg, 25mg-60mg, 27 mg-28mg, 27mg-30mg, 27mg-32mg, 27mg-34mg, 27mg- 36mg, 27mg-38mg, 27mg-40mg, 27mg-42mg, 27mg-44mg, 27mg-46mg, 27mg-48mg, 27mg- 50mg, 27mg-52mg, 27mg-54mg, 27mg-56mg, 27mg-58mg, 27mg-60mg, 30mg-32mg, 30mg- 34mg, 30mg-36mg, 30mg-38mg, 30mg-40mg, 30mg-42mg, 30mg-44mg, 30mg-46mg, 30mg-48mg, 30mg-50mg, 30mg-52mg, 30mg-54mg, 30mg-56mg, 30mg-58mg, 30mg-60mg, 33mg- 34mg, 33mg-36mg, 33mg-38mg, 33mg-40mg, 33mg-42mg, 33mg-44mg, 33mg-46mg, 33mg- 48mg, 33mg-50mg, 33mg-52mg, 33mg-54mg, 33mg-56mg, 33mg-58mg, 33mg-60mg, 36mg- 38mg, 36mg-40mg, 36mg-42mg, 36mg-44mg, 36mg-46mg, 36mg-48mg, 36mg-50mg, 36mg- 52mg, 36mg-54mg, 36mg-56mg, 36mg-58mg, 36mg-60mg, 40mg-42mg, 40mg-44mg, 40mg- 46mg, 40mg-48mg, 40mg-50mg, 40mg-52mg, 40mg-54mg, 40mg-56mg, 40mg-58mg, 40mg- 60mg, 43mg-46mg, 43mg-48mg, 43mg-50mg, 43mg-52mg, 43mg-54mg, 43mg-56mg, 43mg- 58mg, 42mg-60mg, 45mg-48mg, 45mg-50mg, 45mg-52mg, 45mg-54mg, 45mg-56mg, 45mg- 58mg, 45mg-60mg, 48mg-50mg, 48mg-52mg, 48mg-54mg, 48mg-56mg, 48mg-58mg, 48mg- 60mg, 50mg-52mg, 50mg-54mg, 50mg-56mg, 50mg-58mg, 50mg-60mg, 52mg-54mg, 52mg- 56mg, 52mg-58mg, 52mg-60mg, 100mg-200mg, 200mg-300mg, 300mg-400mg, 400mg- 500mg, 50mg-2000 mg, 500mg-1000mg, 1000mg-2000mg, or 1000mg-3000mg . In some embodiments, one or more immune checkpoint inhibitor dose is greater than about 1 mg, 5 mg, about 10 mg, about 12.5 mg, about 13.5 mg, about 15 mg, about 17.5 mg, about 20 mg, about 22.5 mg, about 25 mg, about 27 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, about 150mg, or about 200 mg. In some embodiments, one or more immune checkpoint inhibitor dose is about less than about 5 mg, about 10 mg, about 12.5 mg, about 13.5 mg, about 15 mg, about 17.5 mg, about 20 mg, about 22.5 mg, about 25 mg, about 27 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, about 150mg, or about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 1000 mg, about 2000 mg, or about 3000 mg.

[0161] In some embodiments, the one or more immune checkpoint inhibitor is administered intravenously. In some such embodiments, the one or more immune checkpoint inhibitor is administered over a period of about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, or 240 minutes, or more, or within a range defined by any two of the aforementioned times. In some embodiments, the one or more immune checkpoint inhibitor is administered over a period of about 10 minutes to about 180 minutes, 10 minutes to about 120 minutes, about 20 minutes to about 100 minutes, about 30 minutes to about 90 minutes, or about 30 minutes to about 60 minutes. In some embodiments, the one or more immune checkpoint inhibitor is administered over a period ofabout 30 minutes. In other embodiments, the one or more immune checkpoint inhibitor is administered over a period of about 60 minutes.

[0162] In some embodiments, the one or more immune checkpoint inhibitor is administered 1, 2, 3, 4, or 5 times per treatment cycle. In some embodiments, the one or more immune checkpoint inhibitor is administered 1 time per cycle. In other embodiments, the one or more immune checkpoint inhibitor is administered 2 times per cycle. In some embodiments, wherein the treatment cycle is three weeks (Q3W), the one or more immune checkpoint inhibitor is administered 1 time per cycle. In some such embodiments, one or more immune checkpoint inhibitor is administered on day 1 of the cycle. In some embodiments, wherein the treatment cycle is four weeks (Q4W), the one or more immune checkpoint inhibitor is administered w times per cycle. In some such embodiments, one or more immune checkpoint inhibitor is administered on day 1 and day 15 of the cycle. In some embodiments, the one or more immune checkpoint inhibitor is pembrolizumab, nivolumab, cemiplimab, atezolizumab, avelumab, pidilizumab, ipilimumab, BMS 936559, durvalumab, camrelizumab, dostarlimab, tislelizumab, sintilimab, toripalimab, or a combination thereof. In some embodiments, the one or more immune checkpoint inhibitor is avelumab. In other embodiments, the one or more immune checkpoint inhibitor is atezolizumab. In still other embodiments, the one or more immune checkpoint inhibitor is durvalumab. In yet other embodiments, the one or more immune checkpoint inhibitor is nivolumab. In some embodiments, the one or more immune checkpoint inhibitor is pembrolizumab.

[0163] In some embodiments, plinabulin is administered prior to the administration of one or more immune checkpoint inhibitor. In some embodiments, plinabulin is administered concurrently with one or more immune checkpoint inhibitor. In some embodiments, plinabulin is administered after one or more immune checkpoint inhibitor.

[0164] In some embodiments, plinabulin is administered about 1 min, 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, Ih, 1.5h, 2h, 2.5h, 3h, 4h, 5h, 6h, 7h, 8h, 9h, lOh, l lh, 12h, 13h, 14h, 15h, 16h, 17h, 18h, 19h, 20h, 24h, 30h, 36h, 40h, or 48h, or a range between any two of these values, after the administration of one or more immune checkpoint inhibitor. In some embodiments, plinabulin is administered about 1 min, 5min, 10 min, 15 min, 20 min, 25 min, 30 min, Ih, 1.5h, 2h, 2.5h, 3h, 4h, 5h, 6h, 7h, 8h, 9h, lOh, l lh, 12h, 13h, 14h, 15h, 16h, 17h, 18h, 19h, 20h, 24h, 30h, 36h, 40h, or48h, or a range between any two of these values,before the administration of one or more immune checkpoint inhibitor. In some embodiments, plinabulin is administered in less than about 1 min, 5min, 10 min, 15 min, 20 min, 25 min, 30 min, Ih, 1.5h, 2h, 2.5h, 3h, 4h, 5h, 6h, 7h, 8h, 9h, lOh, l lh, 12h, 13h, 14h, 15h, 16h, 17h, 18h, 19h, 20h, 21h, 22h, 23h, 24h, 30h, 36h, 40h, or 48h, or a range between any two of these values, after the administration of one or more immune checkpoint inhibitor. In some embodiments, plinabulin is administered in more than about 1 min, 5min, 10 min, 15 min, 20 min, 25 min, 30 min, Ih, 1.5h, 2h, 2.5h, 3h, 4h, 5h, 6h, 7h, 8h, 9h, lOh, l lh, 12h, 13h, 14h, 15h, 16h, 17h, 18h, 19h, 20h, 21h, 22h, 23h, 24h30h, 36h, 40h, or 48h, or a range between any two of these values, after the administration of one or more immune checkpoint inhibitor. In some embodiments, plinabulin is administered in less than about 1 min, 5min, 10 min, 15 min, 20 min, 25 min, 30 min, Ih, 1.5h, 2h, 2.5h, 3h, 4h, 5h, 6h, 7h, 8h, 9h, lOh, l lh, 12h, 13h, 14h, 15h, 16h, 17h, 18h, 19h, 20h, 21h, 22h, 23h, 24h, 30h, 36h, 40h, or 48h, or a range between any two of these values, after the administration of one or more immune checkpoint inhibitor. In some embodiments, plinabulin is administered in more than about 1 min, 5min, 10 min, 15 min, 20 min, 25 min, 30 min, Ih, 1.5h, 2h, 2.5h, 3h, 4h, 5h, 6h, 7h, 8h, 9h, lOh, 1 Ih, 12h, 13h, 14h, 15h, 16h, 17h, 18h, 19h, 20h, 21h, 22h, 23h, 24h30h, 36h, 40h, or 48h, or a range between any two of these values, before the administration of one or more immune checkpoint inhibitor. In some embodiments, plinabulin is administered in about lmin-5min, Imin-lOmin, Imin- 15min, lmin-20min, 1 min-25min, 1 min-30min, 0.25h-0.5h, 0.25-0.75h, 0.25-lh,0.5h-lh, 0.5h-2h, 0.5h-2.5h, lh-2h, lh-3h, lh-5h, lh-24h, lmin-24h, or 1 min-2h, 1 day- 2days, Iday - 3days, 1 day-4 days, 1 day-5 days, or 1 day-6 days after the administration of one or more immune checkpoint inhibitor. In some embodiments, plinabulin is administered in about Imin- 5min, Imin-lOmin, lmin-15min, lmin-20min, 1 min-25min, 1 min-30min, 0.25h-0.5h, 0.25- 0.75h, 0.25-lh,0.5h-lh, 0.5h-2h, 0.5h-2.5h, lh-2h, lh-3h, lh-5h, lh-24h, lmin-24h, or 1 min- 2h, 1 day- 2days, Iday - 3 days, 1 day-4 days, 1 day-5 days, or 1 day-6 before the administration of one or more immune checkpoint inhibitor.

[0165] In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin once every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes co-administration of one ormore immune checkpoint inhibitor and plinabulin two times every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin once every 1 week in a treatment cycle of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin twice every 1 week in a treatment cycle of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin on day 1, day 8, and day 15 of a 21-day treatment cycle. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin on day 1 of a 21-day treatment cycle. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin on day 1 and day 4 of a 28-day treatment cycle. In some embodiments, co-administration of one or more immune checkpoint inhibitor and plinabulin includes administering one or more immune checkpoint inhibitor prior to administering plinabulin. In some embodiments, co-administration of one or more immune checkpoint inhibitor plinabulin includes administering one or more immune checkpoint inhibitor after administering plinabulin. In some embodiments, co-administration of one or more immune checkpoint inhibitor and plinabulin includes administering the one or more immune checkpoint inhibitor concurrently with plinabulin. In some embodiments, one or more immune checkpoint inhibitor described in this paragraph can independently be a first, second, third, fourth, fifth, sixth, seventh, or eighth immune checkpoint inhibitor. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin every day of the week for a week. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor plinabulin every day of the week for 2 weeks, 3 weeks, or 4 weeks. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin on day 1 in weekly treatment. In some embodiments, the treatment schedule includes co- administration of one or more immune checkpoint inhibitor and plinabulin on day 1 and day 2 in weekly treatment. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin on day 1, day 2, and day 3 inweekly treatment. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin on day 1, day 2, day 3 in weekly treatment. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin on day 1, day 2, day 3, and day 4 in weekly treatment. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin on day 1, day 2, day 3, day 4, and day 5 in weekly treatment. In some embodiments, the treatment schedule includes co-administration of one or more immune checkpoint inhibitor and plinabulin on day 1, day 2, day 3, day 4, day 5, and day 6 in weekly treatment. In some embodiments, the treatment schedule includes co- administration of one or more immune checkpoint inhibitor composition and plinabulin on day 1, day 3, and day 5 in weekly treatment. In some embodiments, the treatment cycle for plinabulin and the one or more immune checkpoint inhibitors may be the same. In other embodiments, the treatment cycle for plinabulin and the one or more immune checkpoint inhibitors may be different. For example, in some embodiments, the treatment cycle for plinabulin is 21 days, whereas the treatment cycle for the one or more immune checkpoint inhibitors is 14 days. In some embodiments, one or more immune checkpoint inhibitor is used on each administration day can be the same or different. In some embodiments, one or more immune checkpoint inhibitor used on the first administration day is different from one or more immune checkpoint inhibitor used on the rest of the administration days. In some embodiments, one or more immune checkpoint inhibitor used on the first administration day is the same as or different from one or more immune checkpoint inhibitor used on the second administration day. In some embodiments, one or more immune checkpoint inhibitor used on the first administration day is the same as or different from one or more immune checkpoint inhibitor used on the third administration day. In some embodiments, one or more immune checkpoint inhibitor composition used on the first administration day is the same as or different from one or more immune checkpoint inhibitor used on the fourth administration day. In some embodiments, one or more immune checkpoint inhibitor used on the first administration day is the same as or different from one or more immune checkpoint inhibitor used on the fifth administration day. In some embodiments, one or more immune checkpoint inhibitor used on the first administration day is the same as or different from one or more immune checkpoint inhibitor used on the sixth administration day. In some embodiments, one or more immunecheckpoint inhibitor used on the first administration day is the same as or different from one or more immune checkpoint inhibitor used on the seventh administration day.

[0166] In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor (e.g., the first, the second, the third, the fourth, the fifth, the sixth, the seventh, or the eighth) once every 3 weeks. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor once every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor two times every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor once every 1 week in a treatment cycle of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor twice every 1 week in a treatment cycle of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor three times (e.g., day 1, 2, 3, or day 1, 3, 5) every week in a treatment cycle of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor day 1, day 8, and day 15 of a 21-day treatment cycle. The one or more immune checkpoint inhibitor described in this paragraph can independently be the first, second, third, fourth, fifth, sixth, seventh, or eighth one or more immune checkpoint inhibitor. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor every day of the week for a week. In some embodiments, the treatment schedule includes administration of the one or more immune checkpoint inhibitor every day of the week for 2 weeks, 3 weeks, or 4 weeks. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor composition on day 1 in weekly treatment. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor on day 1 and day 2 in weekly treatment. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor on day 1, day 2, and day 3 in weekly treatment. In some embodiments, the treatment schedule includes administration of one or more immunecheckpoint inhibitor on day 1 , day 3, day 5 in weekly treatment. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor on day 1, day 2, day 3, and day 4 in weekly treatment. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor on day 1, day 2, day 3, day 4, and day 5 in weekly treatment. In some embodiments, the treatment schedule includes administration of one or more immune checkpoint inhibitor on day 1, day 2, day 3, day 4, day 5, and day 6 in weekly treatment.

[0167] In some embodiments, the treatment schedule includes administration of plinabulin once every 3 weeks. In some embodiments, the treatment schedule includes administration of plinabulin once every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks,7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes administration of plinabulin two times every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or8 weeks. In some embodiments, the treatment schedule includes administration of plinabulin once every 1 week in a treatment cycle of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes administration of plinabulin twice every 1 week in a treatment cycle of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes administration of plinabulin three times (e.g., day 1, 2, 3, or day 1, 3, 5) every 1 week in a treatment cycle of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatment schedule includes administration of plinabulin on day 1 of a 21-day treatment cycle. In some embodiments, the treatment schedule includes administration of plinabulin on day 1 and day 8 of a 21-day treatment cycle. In some embodiments, the treatment schedule includes administration of plinabulin day 1, day 8, and day 15 of a 21-day treatment cycle. In some embodiments, the treatment schedule includes administration of plinabulin every day of the week for a week. In some embodiments, the treatment schedule includes administration of plinabulin every day of the week for 2 weeks, 3 weeks, or 4 weeks. In some embodiments, the treatment schedule includes administration of plinabulin on day 1 in weekly treatment. In some embodiments, the treatment schedule includes administration of plinabulin on day 1 and day 2 in weekly treatment. In some embodiments, the treatment schedule includes administration of plinabulin on day 1, day 2, and day 3 in weekly treatment. In some embodiments, the treatment schedule includesadministration of plinabulin on day 1 , day 3, day 5 in weekly treatment. In some embodiments, the treatment schedule includes administration of plinabulin on day 1, day 2, day 3, and day 4 in weekly treatment. In some embodiments, the treatment schedule includes administration of plinabulin on day 1, day 2, day 3, day 4, and day 5 in weekly treatment. The treatment schedule includes administration of plinabulin on day 1, day 2, day 3, day 4, day 5, and day 6 in weekly treatment.

[0168] The treatment cycle can be repeated as long as the regimen is clinically tolerated. In some embodiments, the treatment cycle for one or more immune checkpoint inhibitor and plinabulin is repeated for n times, wherein n is an integer in the range of 2 to 30. In some embodiments, n is 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, a new treatment cycle can occur immediately after the completion of the previous treatment cycle. In some embodiments, a new treatment cycle can occur a period of time after the completion of the previous treatment cycle. In some embodiments, a new treatment cycle can occur after 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, or 7 weeks after the completion of the previous treatment cycle.

[0169] Administration of the compositions disclosed herein can be via any of the accepted modes of administration for agents that serve similar utilities including, but not limited to, orally, subcutaneously, buccally, subcutaneously, intravenously, intranasally, intratumorally, topically, transdermally, intraperitoneally, intramuscularly, intrapulmonarilly, vaginally, rectally, intragastrically, or intraocularly. Oral and parenteral administrations are customary in treating the indications that are the subject of some embodiments.

[0170] In some embodiments, the compositions described herein can be used in combination with other therapeutic agents. In some embodiments, the compositions described herein can be administered or used in combination with treatments such as chemotherapy, radiation, and biologic therapies

[0171] To further illustrate this invention, the following examples are included. The examples should not, of course, be construed as specifically limiting the invention. Variations of these examples within the scope of the claims are within the purview of one skilled in the art and are considered to fall within the scope of the invention as described, and claimed herein. The reader will recognize that the skilled artisan, armed with the presentdisclosure, and skill in the art is able to prepare and use the disclosure without exhaustive examples.EXAMPLESExample 1 - In vitro Assays combining plinabulin with radiation

[0172] Radiation can liberate local antigen release, and when coupled to DC maturation, can potentiate systemic immunity with immune checkpoint inhibitors (ICI), even in Id-refractory settings. Here we tested this hypothesis preclinically and clinically in a phase I basket study.

[0173] In vitro assays involved combining plinabulin with radiation in different timings before and after radiation. DC activation was assessed with flow cytometry evaluating CD80, CD86, major histocompatibility complex Class II (MHC-II) expression. TS / A mouse mammary adenocarcinoma cell line (TSA) implanted animals were irradiated with 8 Gy x 3 with or without plinabulin or with or without anti-PD-1 monoclonal antibody.

[0174] SP37A3 and XS 106 DC lines were irradiated with 10 Gy and Plinabulin was added to those cells. DC maturation was enhanced by combining radiation with plinabulin, particularly when radiation was added 3-6 hours prior to plinabulin, but not when plinabulin was added first before radiation. FIG. 1 shows the results of in vitro cell line XS 106 that was treated with plinabulin either 3 hours before irradiation or 1, 3, and 6 hours after irradiation. Results suggest that plinabulin treatment 3 hours after irradiation triggered the greatest activation of DCs. Using TSA syngeneic breast cancer cells in BALB / c mice, tumors were irradiated with 24Gy / 3fr, and treated with aPDl and Plinabulin. There were minimal anti-tumoral effects with aPDl alone, Plinabulin alone, and aPDl+Plinabulin groups. However, triple therapy showed much stronger tumor regression than irradiation and aPDl. The percentage of CD8 T cells and CD86+ DCs in the tumor were significantly increased in the triple combination group that were not seen for the monotherapy or bimodal therapy groups (p<0.05, Dunnett’s test).

[0175] Plinabulin in combination with radiation and immune-checkpoint inhibitors was able to induce systemic immune response in immunotherapy-refractory tumors.Example 2

[0176] This study was an open label, single-center study which assessed the safety and tolerability of plinabulin when administered in combination with radiation / immunotherapy regimens in subjects with one of several metastatic or locally advanced cancers who had disease progression on anti-PD-l / PD-Ll mAb treatment as standard of care, and assessed the objective response rate of the study regimen.

[0177] The study cohort was tumor type specific. There were 8 study cohorts for this study: (1) bladder cancer; (2) melanoma; (3) Merkel cell cancer; (4) MSI-H cancers (of any histology); (5) non-small cell lung cancer (6) renal cell cancer; (7) small cell lung cancer; and (8) any tumor type that has a checkpoint inhibitor approval. The Anti-PD-l / PD-Ll mAb treatment cycle defined the subject’s treatment cycle for the study.

[0178] All subjects received a triple combo treatment of Radiation Therapy (RT) + Plinabulin + anti-PD-l / PD-Ll mAb in Cycle 1, followed by anti-PD-l / PD-Ll mAb and plinabulin combo regimen in Cycle 2 and beyond until disease progression or development of unacceptable toxicity, withdrawal from study treatment, or discontinuation of this study. A short course of local consolidative RT was administered in Cycle 1 starting from Day 1. Optional sequential RT was administered to target other untreated lesions at discretion of the treating doctor in Cycle 2 of any regimens. Plinabulin was dosed on Day 1 and Day 4 of Cycle 1 of any anti-PD-l / PD-Ll regimen, and if optional RT was given in Cycle 2, Plinabulin was also given on Day 4 of Cycle 2. Plinabulin was given on Day 1 of Cycle 3 and thereafter. Anti- PD-l / PD-Ll mAb was dosed on Day 1 of every treatment cycle (also on Day 15 in case of the four week cycle (Q4W) regimen containing Avelumab or Durvalumab or Nivolumab as Anti- PD-l / PD-Ll mAb). Subjects must receive the same anti-PD-l / PD-Ll mAb they failed in the prior treatment.

[0179] _ Radiation therapy: Radiation therapy was delivered using external beam radiation, with either 2D / conventional techniques, three-dimensional conformal therapy, intensity modulated radiation therapy (IMRT), stereotactic radiosurgery (SRS) or proton beam therapy (PBT), at the discretion of the treating radiation oncologist. Radiation Therapy (RT) was administered with one of three regimens: 8 Gy x 3 fractions, 12.5 Gy x 4 fractions, and / or 4 Gy x 5 fractions from Days 1 to 3 (3 fractions), Days 1 to 4 (4 fractions), or Days 1 to 5 (5 fractions) in Cycle 1. The choice of RT regimens for tumors and lesions was at the discretionof the treating radiation oncologist. RT targeted up to a maximum of 5 tumor lesions, and any of the radiation regimens were used simultaneously or sequentially. Optional sequential RT was at the discretion of the treating radiation oncologist if warranted to target other untreated lesions with same regimens described above (Cycle 2 Q4W; Cycle 2 Q3W). Treatment was for any lesions in nodes and organs including brain and bone. At least one measurable lesion was left untreated for disease assessment during the study. Assessment of responses was not reliant on RT treated tumor lesions and bony lesions. Brain metastasis were used for irRECIST response assessment.

[0180] If patients developed toxicity attributable to radiation after receiving at least one dose of radiation, the rest of the radiation treatment was discontinued if deemed by treating radiation oncologist to be in the best interest of the patient, and the AEs were documented. Since the number of fractions was between 3-5 fractions, patients received at least 1 fraction up to 5 fractions. The initially prescribed dose per fraction did change, but it was possible to reduce the total dose delivered. The patient had a visit with the treating radiation oncologist at the end of every cycle of radiation treatment.

[0181] _ Plinabulin Administration. Plinabulin was administered on Day 1 and Day 4 in Cycle 1 intravenously, and if optional RT was given in Cycle 2, Plinabulin was also given on Day 1 and Day 4 of Cycle 2. Optional RT was always given in Cycle 2 on Day 1. Plinabulin was always given on Day 1 of Cycle 3 and after. Two plinabulin dose levels were explored: a dose level at 30 mg / m2was initially tested. If it is not deemed tolerable, then a dose at 20 mg / m2was explored.

[0182] For plinabulin at 30 mg / m2dose level a 60-minute infusion through IV with ± 10 min window was recommended. For plinabulin at 20 mg / m2dose level a 30-minute infusion through IV with ± 5 min window was recommended. For patients with a body surface area (BSA) greater than 2.4 m2, dosing was calculated using the maximum BSA of 2.4 nr for Plinabulin. Plinabulin was administered at 1-2 hours after completion of anti-PD-1 or PD-L1 mAb infusion when applicable, or at least 3 hours (but not longer than 12 hours) after the radiotherapy. The groupings for the study may be found in Table 1.Table 1. Study GroupingsC = cycle, D = day; Q3W = three week cycle; Q4W = four week cycle[01831 Peripheral Blood Collection: Peripheral blood was collected from patients prior to administration of plinabulin on course 1 day 1 (C1D1), course 1 day 4 (C1D4), course 3 day 1 (C3D1), course 5 day 1 (C5D1), and course 7 day 1 (C7D1) as available. The first sample was before any exposure to plinabulin, the second was 3 days after plinabulin infusion and subsequent samples were generally several weeks after the most recent infusion. Blood was generally processed within 4 hours of blood draw.[01841 Peripheral B lood Phenoty in : Fresh heparinized peripheral blood was used for immunophenotyping using 200 pL of whole blood for each panel. Following a 30-minute incubation with the appropriate fluoro phore-conjugated antibody cocktail, whole blood was lysed with BD FACS lyse as per manufacture protocol and washed with PBS. Countbright absolute counting beads (Invitrogen) were added for absolute quantification. Data was acquired with a BD LSR II and analyzed with FlowJo version 9. Dendritic cells were identified with a 4-color gating strategy gating first on all peripheral blood DC as LIN-FITC-negative, HLA-DR-APC-H7 -positive. Plasmacytoid dendritic cells (pDC) were defined as CDl lc- APC-positive CD123-PerPC-negative whereas conventional dendritic cells (eDC) were defined as CD 1 Ic-APC-lo, CD 123-PerCP-positve. Separate tubes were used for different pairs of DC maturation markers after establishing isotype controls from the parental gates. CD86, CD40, and CCR5 were quantified with PE-conjugated antibodies and CD80, CD83 and CCR7were quantified with PE-Cy7 conjugated antibodies. Additional panels were used for lymphocyte and monocyte immunopheno typing.

[0185] Patients with clinical benefits (PR+SD) had significantly higher percentage of CCR7+, CD80+ and CD80+ pDC than patients without clinical benefits (PD). Three days after the very first dose of plinabulin (C1D4), triple combination therapy of radiation therapy / Plinabulin / immune checkpoint inhibitor induced maturation of pDC as highlighted by expression of CCR7, a chemokine receptor that guides mature DC towards lymph nodes, along with increased expression of the co-stimulatory molecules CD80 and CD83. (FIG. 2A). Patients with clinical benefits (PR+SD) had significantly higher percentage of inflammatory monocytes (CD14dimCD16bright). PR+SD patients had a larger increase in the percent of CD14-dim CD16-bright inflammatory monocytes, with a concomitant shift away from a CD14+ CD16- classical monocyte phenotype, whereas PD patients displayed minimal changes in monocyte phenotypes. (FIG. 2B). Overall, the analyses indicate significant or positive trends towards higher expression of maturation markers by pDC (CCR7 / CD80 / CD83) in patients with PR+SD relative to PD patients. The increased CCR7 expression on pDC and monocyte shift from classical to inflammatory phenotype were only seen in PR+SD patients.Example 3 - Computation Analysis for scRNAseq[01861 Mapping and gene counting: Processing of raw reads was performed using the Cell Ranger software provided by 10X genomics and the hg38 human reference genome (GRCm38 v86 and corresponding GENCODE annotation file). Frankish, Adam, et al. "GENCODE reference annotation for the human and mouse genomes." Nucleic acids research 47. DI (2019): D766-D773. During mapping the preprocessing, PCR duplicates were removed. The UMI (unique molecular identifier) for each gene was counted to measure the gene expression levels. The cell doublet status was inferred by the software DoubletFinder. McGinnis, Christopher S., Lyndsay M. Murrow, and Zev J. Gartner. "DoubletFinder: doublet detection in single-cell RNA sequencing data using artificial nearest neighbors." Cell systems 8.4 (2019): 329-337. A number of quality-control metrics were evaluated to control the data quality, including the total UMI count, the total number of genes detected, the mitochondrial gene ratio, logl0(gene number) per UMI, and the inferred status of cell doublet. Low-quality cells with a total UMI count smaller than 500, a total number of genes smallerthan 200, and a mitochondrial gene ratio greater than 50% were excluded for further analysis. Kang, Yunhcc, ct al. "A human forcbrain organoid model of fragile X syndrome exhibits altered neurogenesis and highlights new treatment strategies." Nature neuroscience 24.10 (2021): 1377-1391.[01871 Dimensionality reduction and clustering: Main computational analysis of scRNA-seq read-count matrices was performed using Seurat package (v 4.1.1) in R (v 4.2.1). Satija, Rahul, et al. "Spatial reconstruction of single-cell gene expression data." Nature biotechnology 33.5 (2015): 495-502. After the count matrices and metadata were loaded in Seurat, highly variable genes were identified based on the gene expression variations and PCA was performed for dimension reduction. The top principal components were selected based on the total variance of gene expression they explained, typically using a curated Scree plot approach to catch at least 70% of the total variation. The selected principal components were transformed to t-SNE (T-distributed Stochastic Neighbor Embedding) components or UMAP for visualization. Mclnnes, Leland, John Healy, and James Melville. "Umap: Uniform manifold approximation and projection for dimension reduction." arXiv preprint arXiv: 1802.03426 (2018); Cieslak, Matthew C., et al. "t-Distributed Stochastic Neighbor Embedding (t-SNE): A tool for eco-physiological transcriptomic analysis." Marine genomics 51 (2020): 100723). The created objects were then integrated to a merged object using the canonical correlation analysis (CCA) with the IntegrateData() function. Stuart, Tim, et al. "Comprehensive integration of single-cell data." Cell 177.7 (2019): 1888-1902. Normalization of integrated data was performed, regressing out sample identities using the SCTransform function in Seurat. Hafemeister, Christoph, and Rahul Satija. "Analyzing scRNA-seq data with the setransform and offset models." (2020). Subsequently, the CCA object list were analyzed through a graph-based clustering algorithm via the FindNeighbors() and FindClustersQ functions.

[0188] _ Cell type annotation: A semi-supervised approach was adopted to annotate the cell type information. First, malignant cells were distinguished from non-malignant subsets by applying inferCNV, a widely-used aneuploid cell identification algorithm based on large- scale copy number variations. Second, for the cells that were not labeled as tumor cells, we used the annotated single-cell data from Leader et al. Cancer Cell 39.12 (2021): 1594-1609 as a reference panel and applied scPred to assign the cell type labels in a supervised way.Alquicira-Hernandez, Jose, et al. "scPred: accurate supervised method for cell-type classification from single-cell RNA-scq data." Genome biology 20.1 (2019): 1-17. Supervised cell-type annotation generally achieves more accurate cell-type annotation results than unsupervised methods. All the cell type annotation analysis was performed using the data from each individual separately to adjust for site and disease differences.[01891 Differentially expression and pathway analysis: The differential expressed gene detection was performed by combining all the samples but within each cell type. A generalized linear’ model-based algorithm called MAST, which treats cellular detection rate as a covariate, is applied to detect differentially expressed genes. Finak, Greg, et al. "MAST: a flexible statistical framework for assessing transcriptional changes and characterizing heterogeneity in single-cell RNA sequencing data." Genome biology 16.1 (2015): 1-13. The genes with adjusted p-value smaller than 0.05 were deemed as differential signals. Gene enrichment analysis was performed using clusterProfiler with databases Gene Ontology biological process and KEGG pathways, i.e., Yu, Guangchuang, et al. "clusterProfiler: an R package for comparing biological themes among gene clusters." Omics: a journal of integrative biology 16.5 (2012): 284-287; Ashburner, Michael, et al. "Gene ontology: tool for the unification of biology." Nature genetics 25.1 (2000): 25-29; Kanehisa, Minoru, and Susumu Goto. "KEGG: kyoto encyclopedia of genes and genomes." Nucleic acids research 28.1 (2000): 27-30.

[0190] Single cell RNAseq analysis of non-irradiated tumor biopsies from pretreatment (C1D1) and post-treatment (C3D1) samples of non-irradiated target lesions. A significant increase of Granzyme K+ T-cells (T_GZMK, P = 0.038) was observed in PR+SD subjects while resident memory CD8+ T-cells (CD8 Trm, P = 0.0095), MoMac-III (monocyte- derived macrophage group 3) (P - 0.038), IgA+ plasma (P - 0.023), IgG-i- plasma (P = 0.042), as well as IgM-i- (P = 0.037) plasma cells all significantly decreased in PR+SD subjects (FIG. 3 A and 3B).

[0191] Analysis of tumor-infiltrating DCs for a GEF-H1 -dependent immune signature is shown in FIG. 4A and 4B. The baseline PR+SD subjects had significantly higher GEF-H1 immune activation scores in cDCl, cDC2 and total DCs than PD subjects (FIG. 4A). While GEF-H1 immune activation score increased post-treatment in cDC2, DC3 and total DCs of PR+SD subjects at C3D1, the opposite was observed for the PD subjects (FIG. 4B).

[0192] Single cell RNAseq analysis of tumor biopsies at pre / post triple combination treatment indicates activation of a GEF-H1 -dependent immune signature in monocyte-derived macrophages (MoMac III) (FIG. 5A) in responder patients. Responders were found to have lower levels of GEF-H1 dependent immune signatures in MoMac cells. See Kashyap et al. Cell Rep. 2019 September 24; 28(13): 3367-3380, the disclosure of which is incorporated by reference herein in its entirety, for description of GEF-H1. Further analysis revealed that ARG1 (Arginase 1) expression in MoMac-III was higher in PD subjects than in PR+SD subjects suggesting an M2-biased phenotype for PD patients (FIG. 5B).

[0193] Single-cell RNA sequencing (scRNA-seq) of PBMC was primarily conducted using the Seurat package (version 4.3.0) within R (version 4.3.1). A Seurat object was initialized with the loaded count matrix and associated metadata. The approach for identifying highly variable genes and performing principal component analysis (PCA) was consistent with that used in the scRNA-seq analysis of tumor biopsies. Normalization was also performed using SCTranform function in Seurat. Then, data integration was implemented by harmony algorithm from Harmony package to remove batch effect. UMAP visualization was stratified by patient response categories partial response (PR), stable disease (SD), and progressive disease (PD) as well as by different time points. Lastly, the integrated dataset was subjected to clustering facilitated by the FindNeighbors and FindClusters functions in Seurat.

[0194] Peripheral blood mononuclear cell (PMBC) mean normalized baseline GEF-H1 -dependent immune signature scores were evaluated (FIG. 6A). Responding patients are those showing a partial response (PR); or showing stable disease (SD). Responders (PR + SD) had significantly higher PMBC baseline GEF-H1 scores than non-responders for various cell types, including T cells, NK cells, B cells, monocytes, and dendritic cells. In the PR+SD group, a stable increase trend of GEF-Hl-related gene expression is observed, contrasting with the PD group, where GEF-H1 immune activation score decreases initially from C1D1 to C1D4 and then increases from C1D4 to C3D1. FIG. 6B is the same plot extracted from FIG. 6A, showing the baseline data (at C1D1) correlated with clinical response. FIG. 6C presents the data in FIG. 6A in tabular form.

[0195] Single cell and T-cell receptor sequencing analysis of PBMCs shows the log-fold distribution of cell abundance changes of the small clonal family between C1D1 and C3D1 for PR+SD and PD, respectively (FIG. 7). Each dot represents one cell. For each dot,MiloR collected neighborhood cells and calculated the differential abundance. To compare differential cell abundance between conditions, such as timepoints or response types, a Besswarm plot was generated using the MiloR package. This package randomly collects the cells in the UMAP. For the selected cells, the neighboring cells were collected using K-NN graphs, and the differential cell abundance between the conditions was tested using generalized linear models. The cells having a cell fraction less than 70% were defined as mixed cells. Cells that had a cell fraction of more than 70% kept their original cell types. For newly defined cell types from MiloR, significantly enriched cells were extracted. These cells from the differential cell abundance test had a P- value < 0.01. In the downstream analysis, all plots were created using ggplot2 package v3.4.4 and ggpubr package v0.6.0. Significant compositional differences over time were observed only in the PR+SD patients and in cells of the small clonal family. Specifically, the CD4+ naive, CD4+ central memory T (TCM), CD4+ CTL, regulatory T (Treg), CD8+ naive, CD8+ TCM, and mucosal-associated invariant T cells (MAIT) significantly increase in the PR+SD group at C3D1 compared to CID 1. These changes likely reflect cell proportion changes across time with immune activation, which may relate to the migration pattern of T-lymphocyte subsets in and out of blood circulation during the treatment course.

Claims

WHAT TS CLAIMED TS:

1. A method of treating cancer in a subject, the method comprising a treatment cycle comprising the steps of:(i) administering one or more immune checkpoint inhibitor to the subject;(ii) administering plinabulin or a pharmaceutically acceptable salt thereof to the subject;(iii) obtaining a biological sample from the subject; and(iv) determining the levels of one or more biomarkers or levels of one or more cells in the biological sample.

2. The method of Claim 1, wherein treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the level of the one or more biomarker or one or more cells in the biological sample is above or below a predetermined threshold level, or if a score determined by mathematically combining levels of two or more biomarkers or two or more cells is above or below a predetermined threshold value.

3. The method of Claim 1 or 2, wherein the one or more biomarker is CCR7, CD40, CD80, CD83, CD86, GEF-H1, IL-2, IFNy, IL-6, IL-12p70, IL-12p40, IL-13, IL-17A, IL-23, G-CSF, PD-L1, IL-8 and IFN- , or a combination thereof.

4. The method of Claim 3, wherein the one or more biomarker includes CCR7.

5. The method of Claim 3 or 4, wherein the one or more biomarker includes CD40.

6. The method of any one of Claims 3 to 5, wherein the one or more biomarker includes CD80.

7. The method of any one of Claims 3 to 6, wherein the one or more biomarker includes CD83.

8. The method of any one of Claims 3 to 7, wherein the one or more biomarker includes CD86.

9. The method of any one of Claims 3 to 7, wherein the one or more biomarker includes GEF-H1.

10. The method of any one of Claims 1 to 9, wherein the one or more immune checkpoint inhibitor is pembrolizumab, nivolumab, cemiplimab, atezolizumab, avelumab, pidilizumab, ipilimumab, BMS 936559, durvalumab, camrelizumab, dostarlimab, tislelizumab, sintilimab, toripalimab, or a combination thereof.11 . The method of Claim 10, wherein the one or more immune checkpoint inhibitor is avclumab.

12. The method of Claim 10, wherein the one or more immune checkpoint inhibitor is atezolizumab.

13. The method of Claim 10, wherein the one or more immune checkpoint inhibitor is durvalumab.

14. The method of Claim 10, wherein the one or more immune checkpoint inhibitor is nivolumab.

15. The method of Claim 10, wherein the one or more immune checkpoint inhibitor is pembrolizumab.

16. The method of any one of Claims 1 to 15, wherein the treatment cycle is 21 days.

17. The method of any one of Claims 1 to 15, wherein the treatment cycle is 28 days.

18. The method of any one of Claims 1 to 17, wherein the one or more immune checkpoint inhibitor is administered intravenously.

19. The method of any one of Claims 1 to 18, wherein the one or more immune checkpoint inhibitor is administered on day 1 of the treatment cycle.

20. The method of any one of Claims 1 to 19, wherein the one or more immune checkpoint inhibitor is administered on day 15 of the treatment cycle.

21. The method of any one of Claims 1 to 20, wherein the one or more immune checkpoint inhibitor is administered to the subject in an amount of from 50 mg to 2000 mg22. The method of any one of Claims 1 to 21, wherein the one or more immune checkpoint inhibitor is administered over a period of about 10 minutes to about 180 minutes.

23. The method of Claim 22, wherein the one or more immune checkpoint inhibitor is administered over a period of about 30 minutes to about 90 minutes.

24. The method of Claim 22, wherein the one or more immune checkpoint inhibitor is administered over a period of about 30 minutes.

25. The method of Claim 22, wherein the one or more immune checkpoint inhibitor is administered over a period of about 60 minutes.

26. The method of any one of Claims 1 to 24, wherein the plinabulin is administered on day 1 of the treatment cycle.

27. The method of any one of Claims 1 to 26, wherein the plinabulin is administered on day 4 of the treatment cycle.

28. The method of any one of Claims 1 to 27, wherein the dose of plinabulin is from about 10 mg / m2to about 50 mg / m2.

29. The method of Claim 28, wherein the dose of plinabulin is about 20 mg / m2.

30. The method of Claim 28, wherein the dose of plinabulin is about 30 mg / m2.

31. The method of any one of Claims 1 to 30, wherein the plinabulin is administered over a period of 10 minutes to 180 minutes.

32. The method of Claim 31, wherein the plinabulin is administered over a period of 20 minutes to 90 minutes.

33. The method of Claim 31, wherein the plinabulin is administered over a period of 30 minutes to 60 minutes.

34. The method of any one of Claims 1 to 33, wherein when the one or more immune checkpoint inhibitor and the plinabulin are administered on the same day, the plinabulin is administered from about 0.5 hours to about 3 hours after completion of administration of the one or more immune checkpoint inhibitor.

35. The method of Claim 34, wherein the plinabulin is administered from about 1 hour to about 2 hours after completion of administration of the one or more immune checkpoint inhibitor.

36. The method of any one of Claims 1 to 35, further comprising administering radiation therapy to the subject.

37. The method of Claim 36, wherein radiation therapy is administered in one to ten fractions.

38. The method of Claim 36, wherein radiation therapy is administered three to five fractions.

39. The method of Claim 36, wherein radiation therapy is administered in three fractions.

40. The method of Claim 36, wherein radiation therapy is administered in four fractions.

41. The method of Claim 36, wherein radiation therapy is administered in five fractions.

42. The method any one of Claims 36 to 41, wherein the total dose of radiation administered is from about 1 Gy to about 20 Gy.

43. The method of Claim 42, wherein the total dose of radiation administered is from about 2 Gy to about 15 Gy.

44. The method of Claim 42, wherein the total dose of radiation administered is from about 4 Gy to about 15 Gy.

45. The method of Claim 42, wherein the total dose of radiation administered is about 4 Gy.

46. The method of Claim 42, wherein the total dose of radiation administered is about 8 Gy.

47. The method of Claim 42, wherein the total dose of radiation administered is about 12.5 Gy.

48. The method any one of Claims 36 to 47, wherein radiation therapy is administered on days 1, 2, and 3 of the treatment cycle.

49. The method any one of Claims 36 to 47, wherein radiation therapy is administered on days 1, 2, 3, and 4 of the treatment cycle.

50. The method any one of Claims 36 to 47, wherein radiation therapy is administered on days 1, 2, 3, 4, and 5 of the treatment cycle.

51. The method of any one of Claims 36 to 50, wherein when radiation therapy and plinabulin are administered on the same day, the plinabulin is administered from about 3 hours to about 12 hours after completion of administration of the radiation therapy.

52. The method of any one of Claims 1 to 51, wherein the biological sample is a blood sample.

53. The method of any one of Claims 1 to 51, wherein the biological sample is a tumor biopsy54. The method of any one of Claims 1 to 53, wherein the one or more cells are dendritic cells or their subsets; or wherein the one or more cells are selected from the group consisting of T-cells, CD4+ T-cells CD8+ T-cells, B-cells, IgA-i- plasma cells, IgG-i- plasma cells, and IgM-i- plasma cells; or wherein the one or more cells are selected from the groupconsisting of CD16+ monocytes, CD14+ monocytes, natural killer (NK) cells, granzyme K+ T cells, CD8+ TRM cells, CD8+ TEM cells, CD4+ TCM cells, CD4+ TEM cells, CD4+ naive T-cells, naive B-cells, cDCl cells cDC2 cells, and DC3 cells; or wherein the one or more cells are monocyte-derived macrophage cells (MoMac), or interstitial macrophages (IM); or wherein the one or more cells are immune cells from peripheral blood mononuclear cells (PBMCs).

55. The method of Claim 54, wherein the method comprises determining the level of one or more dendritic cells in the biological sample expressing one or more biomarkers.

56. The method of Claim 54 or 55, wherein the one or more biomarker is CCR7, CD40, CD80, CD83, CD86, GEF-H1, IL-2, IFNy, IL-6, IL-12p70, IL-12p40, IL-13, IL-17A, IL-23, G-CSF, IL-8, IFN- , or a combination thereof.

57. The method of Claim 56, wherein the one or more biomarker is CCR7, CD40, CD80, CD83, CD86, or a combination thereof.

58. The method of Claim 56, wherein the one or more biomarker is GEF-H1.

59. The method of any one of Claims 54 to 58, wherein the dendritic cells are myeloid dendritic cells.

60. The method of any one of Claims 1 to 59, wherein the biological sample obtained on day 4 of the treatment cycle.

61. The method of any one of Claims 1 to 60, wherein the cancer is selected from a breast cancer, a bladder cancer, a glioma, a glioblastoma, a head and neck cancer, a nonsmall cell lung cancer, a small cell lung cancer, recurrent small cell lung cancer (SCLC), a colorectal cancer, a gastrointestinal stromal tumor, a gastroesophageal carcinoma, a renal cell cancer, a prostate cancer, a liver cancer, a colon cancer, a pancreatic cancer, an ovarian cancer, a lymphoma, or a cutaneous T-cell lymphoma, or a melanoma.

62. The method of any one of Claims 1 to 61, comprising determining baseline levels of the one or more biomarkers or one or more cells in an initial biological sample.

63. The method of Claim 62, wherein the initial biological sample is obtained on day 1 of the treatment cycle prior to administration of the one or more immune checkpoint inhibitors and plinabulin.

64. The method of Claim 62 or 63, comprising comparing the levels determined in step (iv) with the baseline levels.

65. The method of Claim 64, wherein the comparing comprises determining a change in the levels of the one or more biomarkers or one or more cells.

66. The method of Claim 65, wherein treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the change in the levels of the one or more biomarkers or one or more cells is below a predetermined threshold.

67. The method of Claim 66, wherein treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the level of pDC expressing CCR7 is greater than about 10 percent below the baseline level.

68. The method of Claim 66, wherein treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the level of pDC expressing CCR7 is greater than about 20 percent below the baseline level.

69. The method of Claim 66, wherein treatment with the one or more immune checkpoint inhibitor and plinabulin is discontinued if the level of pDC expressing CCR7 is less than about 20 percent above the baseline level.

70. A method of treating cancer in a subject, the method comprising a treatment cycle comprising the steps of:(i) obtaining a biological sample from the subject;(ii) determining the levels of one or more biomarkers or levels of one or more cells in the biological sample; and(iii) administering plinabulin or a pharmaceutically acceptable salt thereof to the subject if the levels of one or more biomarkers or levels of one or more cells in the biological sample arc above or below a threshold level, or if a score determined by mathematically combining levels of two or more biomarkers or two or more cells is above or below a predetermined threshold value.

71. The method of Claim 70, wherein the biological sample is a tumor biopsy.

72. The method of Claim 70, wherein the biological sample is peripheral blood.

73. The method of any one of Claims 70 to 72, wherein the one or more cells are dendritic cells or subsets thereof; or wherein the one or more cells are selected from the group consisting of T-cells, CD4+ T-cells CD8+ T-cells, B-cells, IgA-i- plasma cells, IgG-i- plasma cells, and IgM-i- plasma cells; or wherein the one or more cells are selected from the group consisting of CD 16+ monocytes, CD 14+ monocytes, natural killer (NK) cells, granzyme K+T cells, CD8+ TRM cells, CD8+ TEM cells, CD4+ TCM cells, CD4+ TEM cells, CD4+ naive T-cclls, naive B-cclls, cDCl cells cDC2 cells, and DC3 cells; or wherein the one or more cells are monocyte-derived macrophage cells (MoMac), or interstitial macrophages (IM); or wherein the one or more cells are immune cells from peripheral blood mononuclear cells (PBMCs).

74. The method of any one of Claims 70 to 72, wherein the one or more cells are monocyte-derived macrophage cells (MoMac).

75. The method of any one of Claims 70 to 72, wherein the one or more cells are peripheral blood mononuclear cells.

76. The method of any one of Claims 70 to 75, wherein determining the score is a GEF-H1 immune activation score.

77. The method of Claim 76, wherein the GEF-H1 immune activation score is determined using a composite of gene expression levels of one or more genes selected from the group consisting of: Mmpl2, Csfl, Ccl6, Osm, Ccl9, Cxcl3, Clec7a, Hdc, Slcl5a3, Dcstamp, Treml, Cxcl2, Cd300a, Ncf2, Cd80, Lilr4b, Pdcdllg2, Lilrb4a, Mxdl, Spl40. Illa, Ppbp, Gprl71, Ccl7, Il lb, Ccl4, Mania, Aqp9, Creb5, Traf4, Nod2, Ptger2, Mefv, Cd6, Reps2, Tlr7, Sirpblb, Fcgr4, P2ry2, Apol8, Lag3, Pdcdl, Cxcrl, Rab27b, Ctla4, Cxcl9, and Fasl.

78. The method of Claim 76 or 77, wherein the threshold value of GEF-H1 immune activation score is from about 10 to about 40.

79. The method of Claim 76 or 77, wherein the threshold value of GEF-H1 immune activation score is from about 20 to about 40.

80. The method of Claim 76 or 77, wherein the threshold value of GEF-H1 immune activation score is about 30.

81. The method of Claim 76 or 7, wherein the threshold value of GEF-H1 immune activation score is about 25.

82. The method of any one of Claims 70 to 81, wherein the cancer is selected from a breast cancer, a bladder cancer, a glioma, a glioblastoma, a head and neck cancer, a nonsmall cell lung cancer, a small cell lung cancer, recurrent small cell lung cancer (SCLC), a colorectal cancer, a gastrointestinal stromal tumor, a gastroesophageal carcinoma, a renal cell cancer, a prostate cancer, a liver cancer, a colon cancer, a pancreatic cancer, an ovarian cancer, a lymphoma, or a cutaneous T-cell lymphoma, or a melanoma.

83. The method of any one of Claims 70 to 82, further including administration of radiation therapy, an immune checkpoint inhibitor, a chemotherapeutic agent, or a combination thereof, to the subject.

84. The method of any one of Claims 870 to 83, further including administration of radiation therapy and an immune checkpoint inhibitor to the subject.

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