3-Substituted piperidine compounds for Cbl-b inhibition and use of Cbl-b inhibitors in combination with cancer vaccines and / or oncolytic viruses

Abstract

The present invention discloses compounds, compositions and methods for inhibiting the E3 enzyme Cbl-b in the ubiquitin proteasome pathway. The compounds, compositions and methods can be used to regulate the immune system, treat diseases suitable for immune system regulation, and for in vivo, in vitro or ex vivo cell treatment. The present invention also discloses a pharmaceutical composition comprising a Cbl-b inhibitor and a cancer vaccine, and a method for treating cancer using a Cbl-b inhibitor and a cancer vaccine; and a pharmaceutical composition comprising a Cbl-b inhibitor and an oncolytic virus, and a method for treating cancer using a Cbl-b inhibitor and an oncolytic virus.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application Nos. 62 / 831,392, filed April 9, 2019; 62 / 866,909, filed June 26, 2019; 62 / 880,267, filed July 30, 2019; 62 / 888,845, filed August 19, 2019; and 62 / 888,870, filed August 19, 2019, the contents of each of which are incorporated herein by reference in their entirety.

[0003] Technical field of the present invention

[0004] The present invention provides compounds and compositions for inhibiting Cbl-b enzymes and methods of using the compounds and compositions for modulating the immune system, treating diseases, and treating cells in vivo, in vitro, or ex vivo. The present invention also provides pharmaceutical compositions, kits, and methods for treating cancer comprising the combination of the Cbl-b enzyme inhibitor and a cancer vaccine; and pharmaceutical compositions, kits, and methods for treating cancer comprising the combination of the Cbl-b enzyme inhibitor and an oncolytic virus. Background of the Invention

[0006] The ubiquitin-proteasome pathway is a complex system involved in regulating protein function and catabolism. Proteins in eukaryotic cells are conjugated to ubiquitin, a 76-amino acid, 8.5-kilodalton protein. This conjugation, known as ubiquitination, can alter the function or degradation of target proteins. Ubiquitination of target proteins occurs through a series of coupled reactions involving ubiquitin and a group of enzymes known as E1, E2, and E3 enzymes. Ubiquitin is activated by ubiquitin-activating enzymes, or E1 enzymes. Ubiquitin is then transferred to ubiquitin-conjugating enzymes, or E2 enzymes. Finally, ubiquitin ligases, or E3 enzymes, facilitate the transfer of ubiquitin from the E2 enzyme to the target protein. Polyubiquitination of target proteins primarily serves as a signal for the ubiquitin-conjugated protein to be degraded by the proteasome, where it undergoes proteolysis. Ubiquitination by E3 ligases can also lead to changes in protein activity, interactions, or localization. Ubiquitination regulates diverse biological processes, including cell division, DNA repair, and cell signaling.

[0007] Protein synthesis and degradation in cells are crucial for cell cycle regulation, cell proliferation, apoptosis, and many other cellular processes. Therefore, the ability to modulate the ubiquitin-proteasome pathway offers numerous opportunities for intervention in disease processes. Possible mechanisms of intervention include enhancing the degradation of oncogene products, reducing the degradation of tumor suppressor proteins, modulating immune cell responses, and modulating anti-tumor immune responses.

[0008] Therapeutic cancer vaccines have been evaluated in numerous clinical trials. However, only two therapeutic cancer vaccines are approved for use in the United States. Specifically, the Bacillus Calmette-Guérin (BCG) strain of Mycobacterium bovis has been approved for the treatment of bladder cancer, and an ex vivo activated autologous cell vaccine has been approved for the treatment of prostate cancer. Despite this, response rates and overall survival rates for patients receiving cancer vaccines remain far below ideal levels. Therefore, what is needed in the art are methods for improving the efficacy of cancer vaccines.

[0009] Despite numerous clinical trials using oncolytic viruses to treat cancer, only one oncolytic virus has been approved for use in the United States and Europe. Specifically, talimogene laherparepvec, a genetically modified herpes simplex virus, is approved for the treatment of melanoma. However, even talimogene laherparepvec has not been shown to improve overall survival or benefit patients with visceral metastases. Therefore, what is needed in the art are methods to improve the efficacy of oncolytic virus therapy.

[0010] Approximately 35 E2 enzymes and more than 500 E3 enzymes are encoded in the human genome. Casitas B lineage lymphoma proto-oncogene-b (Cbl-b) is an E3 ubiquitin ligase that negatively regulates T cell activation (Wallner et al., Clin Dev Immunol, 2012: 692-639). The discovery of agents that regulate E2 or E3 enzymes provides potential for therapies targeting disease processes involving specific E2 or E3 enzymes. This patent application relates to agents that inhibit one such E3 enzyme (Casitas B lineage lymphoma proto-oncogene-b (Cbl-b)); agents that inhibit Cbl-b used in combination with cancer vaccines, and pharmaceutical compositions comprising Cbl-b inhibitors and cancer vaccines; and agents that inhibit Cbl-b used in combination with oncolytic viruses, and pharmaceutical compositions comprising Cbl-b inhibitors and oncolytic viruses. Abstract

[0011] The present invention discloses compounds and compositions for inhibiting Cbl-b enzymes and methods of using them in vivo, in vitro, or ex vivo to modulate the immune system, treat diseases, and treat cells. The present invention also discloses methods of treating cancer using Cbl-b inhibitors. In short, the Cbl-b inhibitors can be administered to individuals with cancer alone or as part of a combination therapy with one or more of an immune checkpoint inhibitor, an anti-tumor agent, and radiation therapy. In addition, cells treated in vivo and / or in vitro with the compounds or compositions disclosed herein can be used in adoptive cell therapy for treating cancer.

[0012] The present invention discloses a compound represented by formula (I):

[0013]

[0014] or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0015] yes

[0016] X is CH or N;

[0017] Z 1 is CH or N;

[0018] Z 2 is CH or N;

[0019] R 1a and R 1b are independently H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH;

[0020] R 2a is -CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyl-OH, C1-C6 alkyl-CN, or -(C1-C6 alkylene)-O-(C1-C6 alkyl);

[0021] R 2b is H, halogen, or C1-C6 alkyl;

[0022] or R 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro heterocyclic group or a spiro C3-C6 cycloalkyl group consisting of 3 to 6 atoms, wherein at least one of the atoms of the spiro heterocyclic group adjacent to the piperidinyl ring to which it is attached is carbon;

[0023] R 3a and R 3b are independently H, halogen, or C1-C6 alkyl;

[0024] or R 3a and R 3b Together with the carbon atom to which it is attached, it forms a C3-C4 cycloalkyl group;

[0025] R 4 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH;

[0026] yes and Y is CR 5a R 5b or S;

[0027] yes and Y are bonds;

[0028] W is O or a bond;

[0029] R 5a and R 5b are independently H, halogen, or C1-C6 alkyl;

[0030] or R 5a and R 5b Together with the carbon atom to which it is attached, it forms a C3-C6 cycloalkyl group;

[0031] R 6 is a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C6 cycloalkyl group;

[0032] R 7 is H, halogen, C3-C6 cycloalkyl, -NH-(heterocyclyl consisting of 3 to 6 atoms), -NH-(C1-C6 alkyl), -NH-(C3-C6 cycloalkyl), -O-(heterocyclyl consisting of 3 to 6 atoms), -O-(C1-C6 alkyl), or -O-(C3-C6 cycloalkyl);

[0033] R 8 is a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C6 cycloalkyl group;

[0034] R 9 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH; and

[0035] R 10 It is -CF3 or cyclopropyl.

[0036] In some embodiments, yes In some embodiments, Z 1 is CH. In some embodiments, Z 1 It's N.

[0037] In some embodiments, yes

[0038] In some embodiments, yes In some embodiments, Z 2 is CH. In some embodiments, Z 2 It's N.

[0039] In some embodiments, yes

[0040] In some embodiments, yes and Y is CR 5a R 5b or S. In some embodiments, W is a bond or O.

[0041] In some embodiments, yes and Y are bonds.

[0042] In some embodiments, R 8 is C1-C3 alkyl, C1-C3 haloalkyl, or C3-C4 cycloalkyl. 8 It is -CH3, -CF3, or cyclopropyl.

[0043] In some embodiments, X is CH. In some embodiments, X is N.

[0044] In some embodiments, Y is a bond. In some embodiments, Y is CR 5a R 5b In some embodiments, Y is S.

[0045] In some embodiments, R 5a and R 5b are independently H, halogen, or C1-C3 alkyl, or R 5a and R 5b Together with the carbon atom to which it is attached, it forms a C3-C4 cycloalkyl group.

[0046] In some embodiments, R 5a and R 5b are independently H, F, or -CH3; or R 5a and R 5b Together with the carbon atom to which it is attached, it forms a cyclopropyl group.

[0047] In some embodiments, R 5a and R 5b are independently H or F.

[0048] In some embodiments, R 1a and R 1b are independently H, C1-C3 alkyl, C1-C3 haloalkyl, or C1-C3 alkyl-OH. 1a and R 1b are independently H, -CH3, -CF3, or -CH2OH. In some embodiments, R 1b It’s H.

[0049] In some embodiments, R 2a is -CN, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyl-OH, C1-C3 alkyl-CN, or -(C1-C3 alkylene)-O-(C1-C3 alkyl). In some embodiments, R 2a It is -CN, -CH3, -CF3, -CH2OH, -CH2CN, or -CH2-O-CH3.

[0050] In some embodiments, R 2b is H, halogen, or C1-C3 alkyl. 2b It is H, F, or -CH3.

[0051] In some embodiments, R 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro heterocyclic group consisting of 4 to 5 atoms, or a spiro C3-C4 cycloalkyl group. In some embodiments, R 2a and R 2b Together with the carbon atom to which it is attached, it forms a spirocyclopropyl group,

[0052] In some embodiments, R 3a and R 3b are independently H, halogen, or C1-C3 alkyl; or R 3a and R 3b Together with the carbon atom to which it is attached, it forms a C3-C4 cycloalkyl group. 3a and R 3b are independently H, F, or -CH3; or R 3a and R 3b Together with the carbon atom to which it is attached, it forms a cyclopropyl group.

[0053] In some embodiments, R 4 is H, C1-C3 alkyl, C1-C3 haloalkyl, or C1-C3 alkyl-OH. 4 It is H, -CH3, -CF3, or -CH2OH.

[0054] In some embodiments, R 6 is C1-C3 alkyl, C1-C3 haloalkyl, or C3-C4 cycloalkyl. 6 It is -CH3, -CHF2, or cyclopropyl.

[0055] In some embodiments, R 7is H, halogen, C3-C4 cycloalkyl, -NH(heterocyclyl consisting of 4 to 5 atoms), -NH(C1-C3 alkyl), -NH(C3-C5 cycloalkyl), -O(C1-C3 alkyl), -O(heterocyclyl consisting of 4 to 5 atoms), or -O(C3-C5 cycloalkyl). In some embodiments, R 7 is H, Cl, cyclopropyl, -NH(CH2CH3), -NH(cyclopropyl), -OCH2CH3, -O(cyclopropyl),

[0056] In some embodiments, R 9 is H, C1-C3 alkyl, C1-C3 haloalkyl, or C1-C3 alkyl-OH. 9 It is H, -CH3, -CF3, or -CH2OH.

[0057] The present invention also discloses a compound selected from the compounds in Table 1, or a tautomer thereof, or a pharmaceutically acceptable salt thereof. The present invention also discloses a compound selected from any compound disclosed above or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0058] The present invention also discloses a method for regulating the activity of immune cells, comprising contacting the immune cells with an effective amount of any compound disclosed above or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0059] The present invention also discloses a method for treating a cancer responsive to inhibition of Cbl-b activity in an individual in need thereof, comprising administering to the individual an effective amount of any compound disclosed above or disclosed herein, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0060] The present invention also discloses a method for inhibiting Cbl-b activity in an individual in need thereof, comprising administering to the individual an effective amount of any compound disclosed above or disclosed herein, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0061] The present invention also discloses a method for treating or preventing a disease or condition associated with Cbl-b activity in an individual in need thereof, comprising administering to the individual any compound disclosed above or disclosed herein, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0062] The present invention also discloses a method for generating modified immune cells, comprising culturing a cell population comprising immune cells in the presence of an effective amount of any compound disclosed above or disclosed herein, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0063] The present invention also discloses a modified immune cell comprising a Cbl-b inhibitor, wherein the Cbl-b inhibitor is any compound disclosed above or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0064] The present invention also discloses an isolated modified immune cell, wherein the immune cell has been or is contacted with any compound disclosed above or disclosed herein, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0065] The present invention also discloses a composition comprising a cell population comprising isolated modified immune cells, wherein the immune cells have been or are contacted with any compound disclosed above or disclosed herein, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0066] The present invention also discloses a method for inhibiting abnormal cell proliferation, comprising administering an effective amount of isolated modified immune cells to an individual in need thereof, wherein the immune cells have been or are in contact with any compound disclosed above or disclosed herein, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0067] The present invention also discloses a method for inhibiting abnormal cell proliferation, which comprises administering to an individual in need thereof a composition comprising a cell population containing isolated modified immune cells, wherein the immune cells have been or are in contact with any compound disclosed above or disclosed herein, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0068] The present invention also discloses a method for inhibiting abnormal cell proliferation, comprising administering an effective amount of any compound disclosed above or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0069] The present invention also discloses a cell culture composition comprising a cell population comprising immune cells and a Cbl-b inhibitor, wherein the Cbl-b inhibitor is any compound disclosed above or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0070] The present invention also discloses a pharmaceutical composition comprising a Cbl-b inhibitor and one or both of an adjuvant and an antigen, wherein the Cbl-b inhibitor is any compound disclosed above or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0071] The present invention also discloses an article of manufacture comprising any modified immune cell disclosed herein, any composition comprising a cell population disclosed herein, any cell culture composition disclosed herein, or any pharmaceutical composition disclosed herein.

[0072] The present invention also discloses a kit comprising any modified immune cell disclosed in the present invention or any composition comprising the cell population disclosed in the present invention.

[0073] The present invention also discloses the use of a Cbl-b inhibitor in the preparation of a medicament for treating or preventing a disease or condition associated with Cbl-b activity, wherein the Cbl-b inhibitor is any compound disclosed above or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0074] In any embodiment disclosed herein, the Cbl-b protein can be mammalian Cbl-b or human Cbl-b.

[0075] In addition, the present invention discloses Cbl-b inhibitor compounds, vaccines, and compositions comprising Cbl-b inhibitors and vaccines, as well as methods of using the same in treating cancer. The Cbl-b inhibitors and cancer vaccines can be administered to individuals suffering from cancer.

[0076] In addition, the present invention discloses Cbl-b inhibitor compounds, oncolytic viruses, and compositions comprising Cbl-b inhibitors and oncolytic viruses, as well as methods of using the same in treating cancer. The Cbl-b inhibitors and oncolytic viruses can be administered to individuals suffering from cancer.

[0077] The present invention provides a method of immunization, comprising administering to an individual in need thereof an effective amount of a small molecule Cbl-b inhibitor, and administering to the individual an effective amount of a vaccine.

[0078] The present invention provides a method for treating cancer, comprising administering to an individual having cancer an effective amount of a small molecule Cbl-b inhibitor, and administering to the individual an effective amount of an oncolytic virus.

[0079] The present invention provides a method for treating cancer, comprising administering to an individual suffering from cancer an effective amount of an agent capable of lowering the threshold of immune cell activation, and administering to the individual an effective amount of a therapeutic cancer vaccine; or administering to the individual an effective amount of an oncolytic virus.

[0080] The present invention provides a pharmaceutical composition comprising a cancer vaccine and a small molecule Cbl-b inhibitor, optionally wherein the composition further comprises a pharmaceutically acceptable excipient.

[0081] The present invention provides a kit for treating cancer, comprising: (a) a small molecule Cbl-b inhibitor; (b) a therapeutic cancer vaccine; and (c) instructions for administering an effective amount of the Cbl-b inhibitor and the therapeutic cancer vaccine to treat cancer in an individual.

[0082] The present invention provides a kit for treating cancer, comprising: (a) a pharmaceutical composition comprising a small molecule Cbl-b inhibitor and a therapeutic cancer vaccine; and (b) instructions for administering an effective amount of the pharmaceutical composition comprising the Cbl-b inhibitor and the therapeutic cancer vaccine to treat cancer in an individual.

[0083] The present invention provides a pharmaceutical composition comprising an oncolytic virus and a small molecule Cbl-b inhibitor, optionally wherein the composition further comprises a pharmaceutically acceptable excipient.

[0084] The present invention provides a kit for treating cancer, comprising: (a) a small molecule Cbl-b inhibitor; (b) an oncolytic virus; and (c) instructions for administering an effective amount of the Cbl-b inhibitor and the oncolytic virus to treat cancer in an individual.

[0085] Furthermore, the present invention provides a kit for treating cancer, comprising: (a) a pharmaceutical composition comprising a small molecule Cbl-b inhibitor and an oncolytic virus; and (b) instructions for administering an effective amount of the pharmaceutical composition comprising the small molecule Cbl-b inhibitor and the oncolytic virus to treat cancer in an individual.

[0086] BRIEF DESCRIPTION OF THE DRAWINGS

[0087] Figure A Tumor volumes of individual mice are shown.

[0088] Figure B Conditional survival percentages of mice depicted by Kaplan-Meier survival curve plots are shown. Detailed Description of the Invention

[0090] The present invention provides compounds and pharmaceutical compositions that inhibit the Cbl-b enzyme, as well as methods of treating using such compounds and pharmaceutical compositions. The compounds and compositions can be used for methods of regulating the immune system, treating diseases, and treating cells in vivo, in vitro, or ex vivo. In addition, the present invention provides pharmaceutical compositions comprising cancer vaccines and compounds that inhibit the Cbl-b enzyme, as well as methods of treating using such compounds, cancer vaccines, and pharmaceutical compositions. In addition, the present invention also provides pharmaceutical compositions comprising oncolytic viruses and compounds that inhibit the Cbl-b enzyme, as well as methods of treating using such compounds, oncolytic viruses, and pharmaceutical compositions.

[0091] Both T cell activation and T cell tolerance are strictly controlled processes that regulate the immune response to tumors while preventing autoimmunity. Tolerance prevents the immune system from attacking cells expressing "self" antigens. During peripheral tolerance, T cells that recognize "self" antigens (i.e., autoreactive T cells) become functionally unresponsive or deleted after encountering "self" antigens outside the thymus. Therefore, the peripheral tolerance process is important for preventing autoimmune diseases. Normally, cancer cells are cleared by activated T cells that recognize tumor antigens expressed on the surface of cancer cells. However, in cancer, the tumor microenvironment can support T cell tolerance to cancer cells, which enables cancer cells to avoid being recognized and cleared by the immune system. The ability of cancer cells to avoid tumor immune surveillance can lead to uncontrolled tumor growth. Therefore, T cell tolerance may be a form of T cell dysfunction. The general principles of T cell dysfunction are well known in the art (see Schietinger et al., Trends Immunol., 35:51-60, 2014). Other types of T cell dysfunction that can lead to uncontrolled tumor growth include T cell exhaustion, T cell senescence, and / or T cell anergy. Therefore, treating T cell dysfunction, for example, by increasing T cell activation, increasing T cell proliferation, reducing T cell tolerance, and / or reducing T cell exhaustion, can be beneficial for preventing or treating cancer. Other cells of the immune system are important for identifying and eliminating cancer cells during immune surveillance. For example, natural killer (NK) cells are lymphocytes of the innate immune system that can recognize and kill cancer cells (see Martinez-Losato et al., Clin Cancer Res., 21:5048-5056, 2015). Recent studies have also shown that B cell subsets with different phenotypes and functions play different roles in anti-tumor responses (see Saravaria et al., Cell Mol Immunol., 14:662-674, 2017). Due to their role in tumor surveillance, both NK cells and B cells are suitable as therapeutic targets for preventing or treating cancer.

[0092] Cbl-b is a RING-type E3 ligase that plays an important role in the immune system due to its function as a negative regulator of immune activation. Cbl-b plays an important role in reducing T cell activation and thus enhancing T cell tolerance. Studies have found that T cells with Cbl-b deficiency show a lower threshold for activation of antigen recognition receptors and co-stimulatory molecules (e.g., CD28). For example, the absence of Cbl-b in T cells relieves the need for CD28 co-stimulation during T cell activation and proliferation (see Bachmaier et al., Nature, 403: 211-216, 2000). Such cbl-b- / -T cells are largely resistant to T cell anergy, which is a tolerance mechanism in which T cell function is inactivated and T cell proliferation is severely impaired (see Jeon et al., Immunity, 21: 167-177, 2004; and Schwartz et al., Annu Rev Immunol., 21: 305-34, 2003). In support of this, the absence of Cbl-b in cbl-b knockout mice impairs the induction of T cell tolerance and exacerbates autoimmunity (see Jeon et al., Immunity, 21:167-177, 2004). Importantly, the absence of Cbl-b in mice also leads to a robust anti-tumor response that is primarily dependent on cytotoxic T cells. One study demonstrated that cbl-b- / - CD8+ T cells are resistant to T regulatory cell-mediated suppression and exhibit enhanced activation and tumor infiltration. Therapeutic transfer of naive cbl-b- / - CD8+ T cells is sufficient to mediate rejection of established tumors (see Loeser et al., J Exp Med., 204:879-891, 2007). Recent studies have shown that Cbl-b also plays a role in NK cell activation. Genetic deletion of Cbl-b or targeted inactivation of its E3 ligase activity enables NK cells to spontaneously reject metastatic tumors in mouse models (see Paolino et al., Nature, 507:508-512, 2014).

[0093] The present invention provides compounds and compositions that are effective inhibitors of Cbl-b and can be used in novel methods for treating diseases such as cancer. In some embodiments, the compounds and compositions provided herein can be used in methods for modulating the immune system, such as increasing the activation of T cells, NK cells, and B cells, and treating such cells in vivo, in vitro, or ex vivo.

[0094] I. Definition

[0095] An "effective amount" of an agent disclosed herein is an amount sufficient to achieve a specific purpose. An "effective amount" can be determined empirically and routinely based on the purpose. An "effective amount" or "sufficient amount" of an agent is an amount sufficient to produce a desired biological effect (e.g., a beneficial outcome, including a beneficial clinical outcome). In some embodiments, the term "effective amount" refers to an amount of an agent that is effective to "treat" a disease or condition in an individual (e.g., a mammal such as a human).

[0096] As used herein, the term "Cbl-b" refers to a Cbl-b protein. The term also includes naturally occurring Cbl-b variants, including splice variants or allelic variants. The term also includes non-naturally occurring Cbl-b variants, such as recombinant Cbl-b proteins or truncated variants thereof, which generally retain the binding ability of naturally occurring Cbl-b or naturally occurring Cbl-b variants (e.g., the ability to bind to an E2 enzyme).

[0097] The terms "pharmaceutical formulation" and "pharmaceutical composition" refer to a formulation that is in a form that allows the biological activity of the active ingredient to be exerted and does not contain additional components that are unacceptably toxic to the individual to whom the formulation or composition is to be administered. Such formulations or compositions may be sterile. Except for the oncolytic virus, such formulations or compositions may be sterile.

[0098] As used herein, "excipients" include pharmaceutically acceptable excipients, carriers, vehicles, or stabilizers that are non-toxic to cells or mammals exposed thereto at the dosages and concentrations employed. Typically, a physiologically acceptable excipient is a pH buffered aqueous solution.

[0099] Reference to a compound described in a pharmaceutical composition, or a compound described in a claim claiming a pharmaceutical composition, refers to the compound described by the general formula recorded in the pharmaceutical composition, excluding other elements of the pharmaceutical composition, that is, without carriers, excipients, etc.

[0100] The terms "treating" or "managing" a disease refer to the implementation of a regimen that may include administering one or more therapeutic agents to an individual (human or other subject) in an effort to achieve a beneficial or desired outcome, including a clinical outcome, in the individual. Beneficial or desired clinical outcomes include, but are not limited to, alleviation or amelioration of one or more symptoms, alleviation of disease severity, stabilization (i.e., non-worsening) of the disease state, prevention of disease spread, delay or slowing of disease progression, improvement or alleviation of the disease state, and remission (partial or complete). "Treatment" may also refer to prolonging survival compared to the expected survival of an individual who had not received treatment. Furthermore, "treating" and "managing" may occur by administering a single dose of one or more therapeutic agents, or may occur over a series of doses of one or more therapeutic agents. "Treatment" or "managing" does not require complete relief of signs or symptoms, nor does it require a cure. "Treatment" may also refer to clinical intervention, such as administering one or more therapeutic agents to an individual, with the intent to alter the natural course of the disease in the individual or cell being treated (i.e., alter the course of the individual or cell that would occur in the absence of clinical intervention). The term "therapeutic agent" may refer to a Cbl-b inhibitor, a modified immune cell, or a combination thereof.

[0101] As used herein, an "individual" or "subject" is a mammal. "Mammals" for therapeutic purposes include humans; non-human primates; domestic and farm animals; and zoo, sport, or pet animals, such as dogs, horses, rabbits, cows, pigs, hamsters, gerbils, mice, ferrets, rats, cats, and the like. In some embodiments, the individual or subject is a human.

[0102] As used herein, the term "T cell dysfunction" refers to a state of reduced immune responsiveness to antigenic stimulation. The term "T cell dysfunction" encompasses the common elements of T cell exhaustion and / or T cell anergy, in which antigen recognition may occur but the subsequent immune response is ineffective in controlling tumor growth. The term "T cell dysfunction" also encompasses refractory or unresponsiveness to antigen recognition, for example, an impaired ability to translate antigen recognition into downstream T cell effector functions (e.g., proliferation, cytokine production, and / or target cell killing).

[0103] The term "T-cell anergy" refers to a state of unresponsiveness to antigen stimulation due to incomplete or insufficient signaling through the T-cell receptor. T-cell anergy can also occur upon stimulation with antigen in the absence of costimulation, rendering the cell refractory to subsequent activation by the antigen, even in the presence of costimulation.

[0104] The term "T cell exhaustion" refers to a state of T cell dysfunction due to persistent TCR signaling that can occur during cancer. It differs from anergy in that it is not caused by incomplete or defective signaling but rather by persistent signaling. It is defined by poor effector function, persistent expression of inhibitory receptors, and a transcriptional state that differs from that of functional effector or memory T cells.

[0105] "T cell dysfunction" is a disease or condition characterized by reduced responsiveness of T cells to antigenic stimulation. Reduced responsiveness may lead to ineffective control of tumors. In some embodiments, the term "T cell dysfunction" includes cancer, such as hematological cancer or non-hematological cancer. In some embodiments, "T cell dysfunction" is a condition in which T cells are disabled or have a reduced ability to secrete cytokines, proliferate, or perform cytolytic activity.

[0106] "Enhancing / improving T cell function" refers to inducing, causing or stimulating T cells to have a sustained or expanded biological function, or reactivating or reactivating exhausted or inactivated T cells. Examples of enhanced or improved T cell function include increased T cell activation (e.g., increased cytokine production, increased expression of T cell activation markers, etc.), increased T cell proliferation, reduced T cell exhaustion, and / or reduced T cell tolerance compared to the T cell state before treatment with a Cbl-b inhibitor. Methods for determining enhanced or improved T cell function are all known in the art.

[0107] As used herein, "proliferation" refers to the proliferation of cells. Increased proliferation includes the production of a greater number of cells relative to a baseline value. Decreased proliferation includes the production of a decreased number of cells relative to a baseline value. In some embodiments, the cells are immune cells, such as T cells, and increased proliferation is desired. In some embodiments, the cells are cancer cells and decreased proliferation is desired.

[0108] As used herein, "alkyl" refers to a saturated, linear (i.e., unbranched) or branched, monovalent hydrocarbon chain or combinations thereof. Specific alkyl groups are those having a specified number of carbon atoms, for example, alkyl groups having from 1 to 20 carbon atoms ("C1-C2"). 20 alkyl”), having 1 to 10 carbon atoms (“C1-C 10 Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, homologs and isomers such as n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0109] As used herein, "alkenyl" refers to an unsaturated linear (i.e., unbranched) or branched monovalent hydrocarbon chain or combinations thereof having at least one site of olefinic unsaturation (i.e., having at least one moiety of the formula C=C). Specific alkenyl groups are those having the specified number of carbon atoms, for example, having from 2 to 20 carbon atoms ("C2-C 20 alkenyl”), having 2 to 10 carbon atoms (“C2-C 10 Examples of alkenyl groups include, but are not limited to, alkenyl groups having 2 to 8 carbon atoms ("C2-C8 alkenyl"), alkenyl groups having 2 to 6 carbon atoms ("C2-C6 alkenyl"), or alkenyl groups having 2 to 4 carbon atoms ("C2-C4 alkenyl"). Alkenyl groups can be in the "cis" or "trans" configuration, or alternatively, in the "E" or "Z" configuration. Examples of alkenyl groups include, but are not limited to, groups such as ethenyl (or vinyl), prop-1-enyl, prop-2-enyl (or allyl), 2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, buta-1,3-dienyl, 2-methylbuta-1,3-dienyl, homologs and isomers thereof.

[0110] As used herein, "alkynyl" refers to an unsaturated linear (i.e., unbranched) or branched monovalent hydrocarbon chain or combinations thereof having at least one site of acetylenic unsaturation (i.e., having at least one moiety of the formula C≡C). Specific alkynyl groups are those having the specified number of carbon atoms, for example, having from 2 to 20 carbon atoms ("C2-C 20 Alkynyl”), having 2 to 10 carbon atoms (“C2-C 10 Examples of alkynyl groups include, but are not limited to, ethynyl (or ethynyl), prop-1-ynyl, prop-2-ynyl (or propargyl), but-1-ynyl, but-2-ynyl, but-3-ynyl, homologs and isomers thereof.

[0111] As used herein, "alkylene" refers to the same residue as an alkyl group, but with a divalence. Specific alkylene groups are those having 1 to 6 carbon atoms ("C1-C6 alkylene"), 1 to 5 carbon atoms ("C1-C5 alkylene"), 1 to 4 carbon atoms ("C1-C4 alkylene"), or 1 to 3 carbon atoms ("C1-C3 alkylene"). Examples of alkylene groups include, but are not limited to, groups such as methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), butylene (-CH2CH2CH2CH2-).

[0112] As used herein, "cycloalkyl" refers to a non-aromatic, saturated or unsaturated cyclic monovalent hydrocarbon structure. Specific cycloalkyl groups are those having a specified number of ring (i.e., annular) carbon atoms, for example, having 3 to 12 ring carbon atoms ("C3-C 12 A cycloalkyl group is a cycloalkyl group having 3 to 8 ring carbon atoms ("C3-C8 cycloalkyl") or 3 to 6 ring carbon atoms ("C3-C6 cycloalkyl"). A cycloalkyl group may consist of one ring (e.g., cyclohexyl) or multiple rings (e.g., adamantyl), but does not include aromatic (e.g., aryl) groups. Cycloalkyl groups containing more than one ring may be fused, spiro, or bridged, or a combination thereof. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclohexyl ... Cyclobutyl Cyclopentyl Cyclohexyl 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl Norbornyl and similar groups.

[0113] As used herein, "cycloalkylene" refers to the same residues as cycloalkyl, but with a valence of two (i.e., as opposed to a valence of one). Specific cycloalkylene groups are those having from 3 to 12 ring carbon atoms ("C3-C 12 Those having 3 to 8 ring carbon atoms ("C3-C8 cycloalkylene") or those having 3 to 6 ring carbon atoms ("C3-C6 cycloalkylene"). Examples of cycloalkylene include, but are not limited to, cyclopropylene Cyclobutylene Cyclopentylene Cyclohexylene 1,2-cyclohexenylene, 1,3-cyclohexenylene, 1,4-cyclohexenylene, cycloheptylene Norbornylene and similar groups.

[0114] As used herein, "aryl" refers to an aromatic carbocyclic group having a single ring (e.g., phenyl) or multiple fused rings (e.g., naphthyl or anthracenyl), wherein one or more of the fused rings may not be aromatic. Specific aryl groups are those having 6 to 14 ring (i.e., annular) carbon atoms ("C6-C 14 Those having more than one ring (at least one of which is non-aromatic) are attached to the parent structure at an aromatic ring position or a non-aromatic ring position. In one variation, an aryl group having more than one ring (at least one of which is non-aromatic) is attached to the parent structure at an aromatic ring position. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, 1-naphthyl, 2-naphthyl, 1,2,3,4-tetrahydronaphth-6- ... etc.

[0115] "Carbocyclyl" or "carbocycle" refers to an aromatic or non-aromatic monovalent cyclic group in which all ring members are carbon atoms, for example, cyclohexyl, phenyl, 1,2-dihydronaphthyl, and the like.

[0116] As used herein, "arylene" refers to the same residue as aryl, but with a divalence. Specific arylene groups are those having 6 to 14 ring carbon atoms ("C6-C 14 Examples of arylene groups include, but are not limited to, groups such as phenylene, o-phenylene (i.e., 1,2-phenylene), m-phenylene (i.e., 1,3-phenylene), p-phenylene (i.e., 1,4-phenylene), naphthylene, 1,2-naphthylene, 1,3-naphthylene, 1,4-naphthylene, 2,7-naphthylene, 2,6-naphthylene, and the like.

[0117] "Heteroaryl" refers to an unsaturated aromatic cyclic group having from 1 to 14 ring carbon atoms and at least one ring heteroatom, including but not limited to, heteroatoms such as N, O, and S. A heteroaryl group can have a single ring (e.g., pyridyl or imidazolyl) or multiple fused rings (e.g., indolinyl, indolyl, or quinolinyl), wherein at least one of the fused rings is aromatic. Particular heteroaryl groups are 5- to 14-atom rings having 1 to 12 ring carbon atoms and 1 to 6 ring heteroatoms independently selected from the group consisting of nitrogen (N), oxygen (O), and sulfur (S) (“5- to 14-atom heteroaryl”); 5- to 10-atom rings having 1 to 8 ring carbon atoms and 1 to 4 ring heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur (“5- to 10-atom heteroaryl”); or 5-, 6-, or 7-atom rings having 1 to 5 ring carbon atoms and 1 to 4 ring heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur (“5- to 7-atom heteroaryl”). In one variation, heteroaryl includes monocyclic aromatic 5-, 6-, or 7-atom rings having 1 to 6 ring carbon atoms and 1 to 4 ring heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. In another variation, heteroaryl includes polycyclic aromatic rings having 1 to 12 ring carbon atoms and 1 to 6 ring heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Heteroaryl groups having more than one ring, at least one of which is non-aromatic, may be attached to the parent structure at either an aromatic or non-aromatic ring position. Examples of heteroaryl groups include, but are not limited to, groups such as pyridyl, benzimidazolyl, benzotriazolyl, benzo[b]thienyl, quinolinyl, indolyl, benzothiazolyl, and the like. "Heteroaryl" also includes groups such as (2,4-dihydro-3H-1,2,4-triazol-3-one-2-yl) group, which has an aromatic tautomeric structure (1H-1,2,4-Triazol-5-ol-1-yl).

[0118] As used herein, "heterocyclyl" and "heterocyclic group" refer to non-aromatic, saturated or partially unsaturated cyclic groups having a specified number of atoms and heteroatoms, or, if no atoms or heteroatoms are specified, at least 3 ring atoms, 1 to 14 ring carbon atoms, and at least one ring heteroatom, including but not limited to heteroatoms such as N, O, and S. A heterocyclic group can have a single ring (e.g., tetrahydrothienyl, oxazolidinyl) or multiple fused rings (e.g., decahydroquinolinyl, octahydrobenzo[d]oxazolyl). Multiple fused rings include, but are not limited to, bicyclic, tricyclic, and tetracyclic rings, as well as bridged or spirocyclic systems. Examples of heterocyclic groups include, but are not limited to, aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, oxazolidinyl, piperazinyl, morpholinyl, dioxanyl, 3,6-dihydro-2H-pyranyl, 2,3-dihydro-1H-imidazolyl, and the like.

[0119] "Heteroarylene" refers to the same residue as heteroaryl, but with divalence. Specific heteroarylene groups are rings consisting of 5 to 14 atoms having 1 to 12 ring carbon atoms and 1 to 6 ring heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur ("5 to 14-atom heteroarylene"); rings consisting of 5 to 10 atoms having 1 to 8 ring carbon atoms and 1 to 4 ring heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur ("5 to 10-atom heteroarylene"); or rings consisting of 5, 6, or 7 atoms having 1 to 5 ring carbon atoms and 1 to 4 ring heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur ("5 to 7-atom heteroarylene"). Examples of heteroarylene include, but are not limited to, groups such as pyridylene, benzimidazolylene, benzotriazolylene, benzo[b]thienylene, quinolinylene, indolylene, benzothiazolylene.

[0120] "Halo" or "halogen" refers to an element of Group 17 having an atomic number of 9 to 85. Halogen groups include fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0121] “Haloalkyl,” “haloalkylene,” “haloaryl,” “haloarylene,” “haloheteroaryl,” and similar terms refer to a moiety substituted with at least one halogen group. When a haloalkyl moiety or other halogen-substituted moiety is substituted with more than one halogen, this may be indicated by using a prefix corresponding to the number of attached halogen moieties. For example, dihaloaryl, dihaloalkyl, trihaloaryl, trihaloalkyl, etc., refer to aryl and alkyl groups substituted with two (“di”) or three (“tri”) halogen groups, which may be, but not necessarily, the same halogen; thus, for example, the haloaryl group 4-chloro-3-fluorophenyl falls within the scope of dihaloaryl. A subset of haloalkyl groups in which each hydrogen (H) of the alkyl group is replaced by a halogen group is referred to as a “perhaloalkyl.” A specific perhaloalkyl group is trifluoroalkyl (-CF3). Similarly, “perhaloalkoxy” refers to an alkoxy group in which a halogen replaces each hydrogen (H) of the hydrocarbon that constitutes the alkyl moiety of the alkoxy group. An example of a perhaloalkoxy group is trifluoromethoxy (-OCF3). "Haloalkyl" includes monohaloalkyl, dihaloalkyl, trihaloalkyl, perhaloalkyl, and any other number of halogenated substituents that may be present on an alkyl group; the same is true for other groups such as haloalkylene, haloaryl, haloarylene, haloheteroaryl, etc.

[0122] "Amino" refers to the group -NH2.

[0123] "Oxo" refers to the group =0, ie, an oxygen atom double-bonded to carbon or other chemical element.

[0124] Unless otherwise indicated, "optionally substituted" means that the group is unsubstituted or substituted with one or more (e.g., 1, 2, 3, 4, or 5) of the substituents listed for the group, wherein the substituents may be the same or different. In one embodiment, the optionally substituted group is unsubstituted. In one embodiment, the optionally substituted group has one substituent. In another embodiment, the optionally substituted group has two substituents. In another embodiment, the optionally substituted group has three substituents. In another embodiment, the optionally substituted group has four substituents. In some embodiments, the optionally substituted group has 1 to 2, 1 to 3, 1 to 4, or 1 to 5 substituents. When multiple substituents are present, each substituent is independently selected unless otherwise indicated. For example, each (C1-C4 alkyl) substituent on the group -N(C1-C4 alkyl)(C1-C4 alkyl) can be selected independently of one another, thereby generating a group such as -N(CH3)(CH2CH3), and so on.

[0125] In addition to the present disclosure, the term "substituted" when used to modify a specific group or atomic group can also mean that one or more hydrogen atoms (H) of the specific group or atomic group are independently replaced by the same or different substituents as defined herein. In some embodiments, the substituted group has 1, 2, 3 or 4 substituents, 1, 2 or 3 substituents, 1 or 2 substituents, or 1 substituent.

[0126] Unless otherwise indicated, a substituent may be attached to any chemically possible position on the designated group or radical. Thus, for example, in one embodiment, -C1-C8 alkyl-OH includes, for example, -CH2CH2OH and -CH(OH)-CH3, and -CH2C(OH)(CH3)2, and the like. As a further example, in one embodiment, -C1-C6 alkyl-OH includes, for example, -CH2CH2OH and -CH(OH)-CH3, and -CH2C(OH)(CH3)2, and the like. As a further example, in one embodiment, -C1-C6 alkyl-CN includes, for example, -CH2CH2CN and -CH(CN)-CH3, and -CH2C(CN)(CH3)2, and the like.

[0127] Unless a specific isotope of an element is indicated in a general formula, the present disclosure includes all isotopologues of the compounds disclosed herein, such as, for example, deuterated derivatives of the compounds (wherein H may be 2H, i.e., deuterium (D)). Deuterated compounds may provide favorable changes in pharmacokinetic (ADME) properties. Isotopologues may have isotopic substitutions at any or all positions in the structure, or may have atoms present at natural abundance at any or all positions in the structure.

[0128] As used herein, "small molecule" refers to a compound having a molecular weight of 1,000 Daltons or less.

[0129] A hydrogen atom may also be replaced by a close bioisostere, such as fluorine, as long as such replacement results in a stable compound.

[0130] The present disclosure also includes any or all stereochemical forms, including any enantiomeric or diastereomeric forms of the compounds described herein, as well as cis / trans or E / Z isomers. Unless stereochemistry is explicitly indicated in a chemical structure or name, the structure or name is intended to include all possible stereoisomers of the described compound. In addition, when a specific stereochemical form is described, it should be understood that all other stereochemical forms, as well as general non-stereospecific forms of the disclosed compounds and mixtures in any ratio, including mixtures of two or more stereochemical forms of the disclosed compounds in any ratio, such that racemic, non-racemic, enantiomerically enriched, and scaled mixtures of the compounds are also described and included in the present invention. Compositions comprising the disclosed compounds, such as compositions of substantially pure compounds (including their specific stereochemical forms), are also contemplated. Compositions comprising mixtures of the disclosed compounds in any ratio are also included in the present disclosure, including compositions comprising mixtures of two or more stereochemical forms of the disclosed compounds in any ratio, such that racemic, non-racemic, enantiomerically enriched, and scaled mixtures of the compounds are all included in the present invention. If the stereochemistry is specified for one or more parts of a molecule, but not for another part or parts of the molecule, the structure is intended to encompass all possible stereoisomers of the part or parts for which the stereochemistry is not specified. The present invention also includes any and all tautomeric forms of the compounds described herein.

[0131] The present invention is intended to include all salts of the compounds described herein, as well as methods of using such salts of the compounds. In one embodiment, the salts of the compounds include pharmaceutically acceptable salts. Pharmaceutically acceptable salts are those salts that can be administered as medicines or drugs to humans and / or animals and that retain at least some of the biological activity of the free compound (neutral compound or non-salt compound) when administered. The desired salts of the basic compound can be prepared by treating the compound with an acid by methods known to those skilled in the art. Examples of inorganic acids include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Examples of organic acids include, but are not limited to, formic acid, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, sulfonic acid, and salicylic acid. Salts of the basic compound with amino acids, such as aspartate and glutamate, can also be prepared. The desired salts of the acidic compound can be prepared by treating the compound with a base by methods known to those skilled in the art. Examples of inorganic salts of acid compounds include, but are not limited to, alkali metal and alkaline earth metal salts, such as sodium, potassium, magnesium, and calcium salts; ammonium salts; and aluminum salts. Examples of organic salts of acid compounds include, but are not limited to, salts of procaine, dibenzylamine, N-ethylpiperidine, N,N'-dibenzylethylenediamine, and triethylamine. Salts of acid compounds with amino acids, such as lysine salts, can also be prepared. For a list of pharmaceutically acceptable salts, see, for example, P.H. Stahl and C.G. Wermuth (eds.) "Handbook of Pharmaceutical Salts, Properties, Selection and Use" Wiley-VCH, 2011 (ISBN: 978-3-90639-051-2). Several pharmaceutically acceptable salts are also disclosed in the following literature: Berge, J. Pharm. Sci. 66:1 (1977).

[0132] The IC for Cbl-b inhibition was determined as described in Biological Examples 1, 8, and / or 12. 50 The assay for determining the Cbl-b activity with a specific value (Cbl-b inhibition assay) uses a mixture comprising N-terminally biotinylated Avi-tagged Cbl-b, a fluorescently labeled inhibitor probe labeled with BODIPY FL (Example 46), and assay buffer. In one embodiment, the assay for determining the IC value for inhibiting Cbl-b is as follows: 50 The Cbl-b activity assay (Cbl-b inhibition assay) was performed using the conditions described in Biological Examples 1, 8, and / or 12, using 0.5 nM Cbl-b ("high" final concentration). In another embodiment, the IC for inhibition of Cbl-b was determined. 50Cbl-b activity was determined (Cbl-b inhibition assay) using the conditions described in Biological Examples 1, 8, and / or 12, employing 0.125 nM Cbl-b ("low" final concentration).

[0133] It should be understood that certain features of the present disclosure described in the context of separate embodiments for the sake of clarity may also be provided in combination in a single embodiment. Conversely, various features of the present disclosure described in the context of a single embodiment may also be provided individually or in any suitable subcombination for the sake of brevity. All combinations of embodiments relating to the chemical groups represented by the variables are specifically encompassed by the present invention and disclosed herein, just as each combination is individually and explicitly disclosed, to the extent that such combinations include stable compounds (i.e., compounds that can be isolated, characterized, and tested for biological activity). In addition, all subcombinations of chemical groups listed in the embodiments describing such variables are also specifically encompassed by the present invention and disclosed herein, just as each such subcombination of chemical groups is individually and explicitly disclosed herein.

[0134] It is to be understood that aspects (cases) and embodiments of the present invention described as "comprising" or "including" or "covering" include "consisting of" and "consisting essentially of" embodiments.

[0135] The singular forms "a", "an", and "the" used in the present invention and the appended claims include plural forms unless otherwise specified or the context makes clear. For example, "a" excipient or "the" excipient includes one or more excipients.

[0136] Reference to "about" a value includes 90% to 110% of the value. For example, about 50 billion cells refers to 45 to 55 billion cells, including 50 billion cells. For example, a temperature of "about 100 degrees" refers to a temperature of about 90 degrees to about 110 degrees.

[0137] When a numerical range of compounds is given, all compounds within the numerical range, including any designated "a" and "b," are included unless specifically excluded. For example, a reference to compounds 41-43 refers to compound 41, compound 42, and compound 43.

[0138] II.Compounds

[0139] In one aspect, the present invention provides a compound represented by formula (I):

[0140]

[0141] or a tautomer thereof, or a pharmaceutically acceptable salt thereof,

[0142] in:

[0143] yes

[0144] X is CH or N;

[0145] Z 1 is CH or N;

[0146] Z 2 is CH or N;

[0147] R 1a and R 1b are independently H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH;

[0148] R 2a is -CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyl-OH, C1-C6 alkyl-CN, or -(C1-C6 alkylene)-O-(C1-C6 alkyl);

[0149] R 2b is H, halogen, or C1-C6 alkyl;

[0150] or R 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro heterocyclic group or a spiro C3-C6 cycloalkyl group consisting of 3 to 6 atoms, wherein at least one of the atoms of the spiro heterocyclic group adjacent to the piperidinyl ring to which it is attached is carbon;

[0151] R 3a and R 3b are independently H, halogen, or C1-C6 alkyl;

[0152] or R 3a and R 3b Together with the carbon atom to which it is attached, it forms a C3-C4 cycloalkyl group;

[0153] R 4 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH;

[0154] yes and Y is CR 5a R 5b or S;

[0155] yes and Y are bonds;

[0156] W is O or a bond;

[0157] R5a and R 5b are independently H, halogen, or C1-C6 alkyl;

[0158] or R 5a and R 5b Together with the carbon atom to which it is attached, it forms a C3-C6 cycloalkyl group;

[0159] R 6 is a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C6 cycloalkyl group;

[0160] R 7 is H, halogen, C3-C6 cycloalkyl, -NH-(heterocyclyl consisting of 3 to 6 atoms), -NH-(C1-C6 alkyl), -NH-(C3-C6 cycloalkyl), -O-(heterocyclyl consisting of 3 to 6 atoms), -O-(C1-C6 alkyl), or -O-(C3-C6 cycloalkyl);

[0161] R 8 is a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C6 cycloalkyl group;

[0162] R 9 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH; and

[0163] R 10 It is -CF3 or cyclopropyl.

[0164] In another aspect, the present invention provides a compound represented by formula (Ia):

[0165]

[0166] or a tautomer thereof, or a pharmaceutically acceptable salt thereof,

[0167] in:

[0168] yes

[0169] X is CH or N;

[0170] Z 1 is CH or N;

[0171] Z 2 is CH or N;

[0172] R 1a and R 1b are independently H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH;

[0173] R2a is -CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyl-OH, C1-C6 alkyl-CN, or -(C1-C6 alkylene)-O-(C1-C6 alkyl);

[0174] R 2b is H, halogen, or C1-C6 alkyl;

[0175] or R 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro heterocyclic group or a spiro C3-C6 cycloalkyl group consisting of 3 to 6 atoms, wherein at least one of the atoms of the spiro heterocyclic group adjacent to the piperidinyl ring to which it is attached is carbon;

[0176] R 3a and R 3b are independently H, halogen, or C1-C6 alkyl;

[0177] or R 3a and R 3b Together with the carbon atom to which it is attached, it forms a C3-C4 cycloalkyl group;

[0178] R 4 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH;

[0179] yes and Y is CR 5a R 5b or S;

[0180] yes and Y are bonds;

[0181] W is O or a bond;

[0182] R 5a and R 5b are independently H, halogen, or C1-C6 alkyl;

[0183] or R 5a and R 5b Together with the carbon atom to which it is attached, it forms a C3-C6 cycloalkyl group;

[0184] R 6 is a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C6 cycloalkyl group;

[0185] R 7is H, C3-C6 cycloalkyl, -NH-(heterocyclyl consisting of 3 to 6 atoms), -NH-(C1-C6 alkyl), -NH-(C3-C6 cycloalkyl), -O-(heterocyclyl consisting of 3 to 6 atoms), -O-(C1-C6 alkyl), or -O-(C3-C6 cycloalkyl);

[0186] R 8 is a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C6 cycloalkyl group;

[0187] R 9 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH; and

[0188] R 10 It is -CF3 or cyclopropyl.

[0189] In some embodiments, (i.e., the ring A group portion), is In some embodiments, Z 1 is CH. In other embodiments, Z 1 is N. In some embodiments, the Ring A group moiety is In some embodiments, Z 2 is CH. In other embodiments, Z 2 is N. In some embodiments, R 10 In other embodiments, R 10 In some embodiments, the ring A group is selected from the group consisting of: In some embodiments, the Ring A group moiety is In some embodiments, the Ring A group moiety is In some embodiments, the Ring A group moiety is In some embodiments, the Ring A group moiety is In some embodiments, the Ring A group moiety is In some embodiments, the Ring A group moiety is In some embodiments, the Ring A group moiety is In some embodiments, the Ring A group moiety is

[0190] In some embodiments, R 1a and R 1b are independently H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH. 1a and R 1bare independently H, C1-C3 alkyl, C1-C3 haloalkyl, or C1-C3 alkyl-OH. 1a and R 1b are independently H, -CH3, -CF3, or -CH2OH. In some embodiments, R 1a is H. In some embodiments, R 1b is H. In some embodiments, R 1a and R 1b are H. In some embodiments, R 1a and R 1b One of them is H, and the other is C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH.

[0191] In some embodiments, R 1a and R 1b are independently C1-C6 alkyl. 1a and R 1b are independently C1-C3 alkyl. 1a and R 1b are independently methyl, ethyl, n-propyl, or isopropyl. 1a and R 1b In some embodiments, R 1a and R 1b One of them is H and the other is methyl.

[0192] In some embodiments, R 1a and R 1b are independently C1-C6 haloalkyl. 1a and R 1b are independently C1-C6 haloalkyl groups containing 1-13 halogen atoms. 1a and R 1b are independently C1-C6 haloalkyl groups containing 1-7 halogen atoms. 1a and R 1b are independently C1-C3 haloalkyl. 1a and R 1b are independently C1-C3 haloalkyl groups containing 1-7 halogen atoms. 1a and R 1bare each independently a C1-C2 haloalkyl group containing 1-5 halogen atoms. In some embodiments, the halogen atoms are each independently selected from the group consisting of chlorine atoms, bromine atoms, and fluorine atoms. In some embodiments, the halogen atoms are each independently selected from the group consisting of chlorine atoms and fluorine atoms. In some embodiments, the halogen atoms are all fluorine atoms. In some embodiments, the halogen atoms are a combination of chlorine atoms and fluorine atoms. In some embodiments, R 1a and R 1b are independently -CF3, -CCl3, -CF2Cl, -CFCl2, -CHF2, -CH2F, -CHCl2, -CH2F, or -CHFCl. 1a and R 1b In some embodiments, R 1a and R 1b One of them is H and the other is -CF3.

[0193] In some embodiments, R 1a and R 1b are independently C1-C6 alkyl-OH. 1a and R 1b are independently C1-C3 alkyl-OH. 1a and R 1b are independently -CH2OH, -CH2CH2-OH, -CH2CH2CH2-OH, or -C(CH3)2-OH. In some embodiments, R 1a and R 1b In some embodiments, R 1a and R 1b One of them is H and the other is -CH2OH.

[0194] The compound represented by formula (I) corresponds to the substituent R 2a and R 2b In some embodiments, the compound of formula (I) comprises a carbon atom directly connected to the piperidinyl ring at at least one position corresponding to the substituent R 2a and R 2b The two positions of contain carbon atoms directly attached to the linked piperidinyl ring.

[0195] In some embodiments, R 2a is -CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyl-OH, C1-C6 alkyl-CN, or -(C1-C6 alkylene)-O-(C1-C6 alkyl). In some embodiments, R 2ais -CN, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyl-OH, C1-C3 alkyl-CN, or -(C1-C3 alkylene)-O-(C1-C3 alkyl). In some embodiments, R 2a It is -CN, -CH3, -CF3, -CH2OH, -CH2CN, or -CH2-O-CH3.

[0196] In some embodiments, R 2a It is -CN.

[0197] In some embodiments, R 2a is C1-C6 alkyl. In some embodiments, R 2a is C1-C3 alkyl. In some embodiments, R 2a is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 2a It is -CH3.

[0198] In some embodiments, R 2a is C1-C6 haloalkyl. 2a is a C1-C6 haloalkyl group containing 1 to 13 halogen atoms. 2a is a C1-C6 haloalkyl group containing 1-7 halogen atoms. 2a is C1-C3 haloalkyl. 2a is a C1-C3 haloalkyl group containing 1-7 halogen atoms. 2a is a C1-C2 haloalkyl group containing 1-5 halogen atoms. In some embodiments, the halogen atoms are independently selected from the group consisting of chlorine atoms, bromine atoms, and fluorine atoms. In some embodiments, the halogen atoms are independently selected from the group consisting of chlorine atoms and fluorine atoms. In some embodiments, the halogen atoms are all fluorine atoms. In some embodiments, the halogen atoms are a combination of chlorine atoms and fluorine atoms. In some embodiments, R 2a is -CF3, -CCl3, -CF2Cl, -CFCl2, -CHF2, -CH2F, -CHCl2, -CH2F, or -CHFCl. In some embodiments, R 2a Yes -CF3.

[0199] In some embodiments, R 2a is C1-C6 alkyl-OH. In some embodiments, R 2a is C1-C3 alkyl-OH. In some embodiments, R 2ais -CH2OH, -CH2CH2-OH, -CH2CH2CH2-OH, or -C(CH3)2-OH. In some embodiments, R 2a It is -CH2OH.

[0200] In some embodiments, R 2a is C1-C6 alkyl-CN. In some embodiments, R 2a is C1-C3 alkyl-CN. In some embodiments, R 2a is -CH2CN, -CH2CH2-CN, -CH2CH2CH2-CN, or -C(CH3)2-CN. In some embodiments, R 2a It is -CH2CN.

[0201] In some embodiments, R 2a is -(C1-C6 alkylene)-O-(C1-C6 alkyl). In some embodiments, R 2a is -(C1-C3 alkylene)-O-(C1-C3 alkyl). In some embodiments, R 2a is -(C1-C2 alkylene)-O-(C1-C2 alkyl). In some embodiments, R 2a In some embodiments, R 2a It is -CH2-O-CH3.

[0202] In some embodiments, R 2b is H, halogen, or C1-C6 alkyl. 2b is H, halogen, or C1-C3 alkyl. 2b It is H, F, or -CH3.

[0203] In some embodiments, R 2b It’s H.

[0204] In some embodiments, R 2b is halogen. In some embodiments, R 2b is chlorine, fluorine, or bromine. In some embodiments, R 2b is chlorine or fluorine. In some embodiments, R 2b It's fluorine.

[0205] In some embodiments, R 2b is C1-C6 alkyl. In some embodiments, R 2b is C1-C3 alkyl. In some embodiments, R 2bis methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 2b It is -CH3.

[0206] In some embodiments, R 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro heterocyclic group consisting of 3 to 6 atoms or a spiro C3-C6 cycloalkyl group, wherein at least one of the atoms of the spiro heterocyclic group adjacent to the piperidinyl ring to which it is attached is carbon. In some embodiments, R 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro 4 to 5-atom heterocyclic group or a spiro C3-C4 cycloalkyl group. 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro C3-C4 cycloalkyl. 2a and R 2b Together with the carbon atom to which it is attached, it forms a spirocyclopropyl or spirocyclobutyl group. 2a and R 2b Together with the carbon atom to which it is attached, it forms a spirocyclopropyl group. 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro heterocyclic group consisting of 4 to 5 atoms. 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro 4 to 5-atom heterocyclic group containing 1-3 heteroatoms selected from the group consisting of O, N and S. In some embodiments, R 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro 4 to 5-atom heterocyclic group containing 1-3 N atoms and / or O atoms. 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro 4 to 5-atom heterocyclic group containing 1-2 O atoms. 2a and R 2b Together with the carbon atom to which it is attached, it forms In some embodiments, R 2a and R 2b Together with the carbon atom to which it is attached, it forms In some embodiments, R 2a and R 2b Together with the carbon atom to which it is attached, it forms In some embodiments, R 2a and R 2b Together with the carbon atom to which it is attached, it forms

[0207] In some embodiments, R 3a and R 3b are independently H, halogen, or C1-C6 alkyl. 3a and R 3b are independently H, halogen, or C1-C3 alkyl. 3a and R 3b are independently H, F, or -CH3.

[0208] In some embodiments, R 3a is H. In some embodiments, R 3b is H. In some embodiments, R 3a and R 3b are H. In some embodiments, R 3a and R 3b One of them is H, and the other is halogen or C1-C6 alkyl.

[0209] In some embodiments, R 3a and R 3b are each independently halogen. In some embodiments, R 3a and R 3b are independently chlorine, fluorine, or bromine. 3a and R 3b are independently chlorine or fluorine. 3a In some embodiments, R 3b In some embodiments, R 3a and R 3b In some embodiments, R 3a and R 3b All are fluorine.

[0210] In some embodiments, R 3a and R 3b are independently C1-C6 alkyl. 3a and R 3b are independently C1-C3 alkyl. 3a and R 3b are independently methyl, ethyl, n-propyl, or isopropyl. 3a and R 3b In some embodiments, R 3a and R 3b One of them is -CH3.

[0211] In some embodiments, R3a and R 3b Together with the carbon atom to which it is attached, it forms a C3-C4 cycloalkyl group. 3a and R 3b Together with the carbon atom to which it is attached, it forms a cyclopropyl group. 3a and R 3b Together with the carbon atom to which it is attached, it forms a cyclobutyl group.

[0212] In some embodiments, R 4 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH. 4 is H, C1-C3 alkyl, C1-C3 haloalkyl, or C1-C3 alkyl-OH. 4 It is H, -CH3, -CF3, or -CH2OH.

[0213] In some embodiments, R 4 It’s H.

[0214] In some embodiments, R 4 is C1-C6 alkyl. In some embodiments, R 4 is C1-C3 alkyl. In some embodiments, R 4 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 4 It is -CH3.

[0215] In some embodiments, R 4 is C1-C6 haloalkyl. 4 is a C1-C6 haloalkyl group containing 1 to 13 halogen atoms. 4 is a C1-C6 haloalkyl group containing 1-7 halogen atoms. 4 is C1-C3 haloalkyl. 4 is a C1-C3 haloalkyl group containing 1-7 halogen atoms. 4 is a C1-C2 haloalkyl group containing 1-5 halogen atoms. In some embodiments, the halogen atoms are independently selected from the group consisting of chlorine atoms, bromine atoms, and fluorine atoms. In some embodiments, the halogen atoms are independently selected from the group consisting of chlorine atoms and fluorine atoms. In some embodiments, the halogen atoms are all fluorine atoms. In some embodiments, the halogen atoms are a combination of chlorine atoms and fluorine atoms. In some embodiments, R 4is -CF3, -CCl3, -CF2Cl, -CFCl2, -CHF2, -CH2F, -CHCl2, -CH2F, or -CHFCl. In some embodiments, R 4 Yes -CF3.

[0216] In some embodiments, R 4 is C1-C6 alkyl-OH. In some embodiments, R 4 is C1-C3 alkyl-OH. In some embodiments, R 4 is -CH2OH, -CH2CH2-OH, -CH2CH2CH2-OH, or -C(CH3)2-OH. In some embodiments, R 4 It is -CH2OH.

[0217] In some embodiments, X is CH. In other embodiments, X is N.

[0218] In some embodiments, (i.e., the ring D group portion) is In some embodiments, W is O, and the ring D group moiety is In other embodiments, W is a bond, and the ring D group moiety is In some embodiments, the ring D group moiety is In some embodiments, the ring D group moiety is

[0219] In some embodiments, the ring D group moiety is and R 8 is C1-C6 alkyl, C1-C6 haloalkyl, or C3-C6 cycloalkyl. 8 is C1-C3 alkyl, C1-C3 haloalkyl, or C3-C4 cycloalkyl. 8 Is -CH3, -CF3, or cyclopropyl. In some embodiments, the ring D group portion is

[0220] In some embodiments, R 8 is C1-C6 alkyl. In some embodiments, R 8 is C1-C3 alkyl. In some embodiments, R 8 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 8 It is -CH3.

[0221] In some embodiments, R 8 is C1-C6 haloalkyl. 8is a C1-C6 haloalkyl group containing 1 to 13 halogen atoms. 8 is a C1-C6 haloalkyl group containing 1-7 halogen atoms. 8 is C1-C3 haloalkyl. 8 is a C1-C3 haloalkyl group containing 1-7 halogen atoms. 8 is a C1-C2 haloalkyl group containing 1-5 halogen atoms. In some embodiments, the halogen atoms are independently selected from the group consisting of chlorine atoms, bromine atoms, and fluorine atoms. In some embodiments, the halogen atoms are independently selected from the group consisting of chlorine atoms and fluorine atoms. In some embodiments, the halogen atoms are all fluorine atoms. In some embodiments, the halogen atoms are a combination of chlorine atoms and fluorine atoms. In some embodiments, R 8 is -CF3, -CCl3, -CF2Cl, -CFCl2, -CHF2, -CH2F, -CHCl2, -CH2F, or -CHFCl. In some embodiments, R 8 Yes -CF3.

[0222] In some embodiments, R 8 is a C3-C4 cycloalkyl group. 8 In some embodiments, R 8 It is cyclobutyl.

[0223] In some embodiments, Y is S. In other embodiments, Y is a bond. In further embodiments, Y is CR 5a R 5b , where R 5a and R 5b are independently H, halogen, or C1-C6 alkyl, or R 5a and R 5b Together with the carbon atom to which it is attached, it forms a C3-C6 cycloalkyl group.

[0224] In some embodiments, R 5a and R 5b are independently H, halogen, or C1-C6 alkyl. 5a and R 5b are independently H, halogen, or C1-C3 alkyl. 5a and R 5b are independently H, F, or -CH3.

[0225] In some embodiments, R 5a is H. In some embodiments, R 5bis H. In some embodiments, R 5a and R 5b are H. In some embodiments, R 5a and R 5b One of them is H and the other is halogen or C1-C6 alkyl. In some embodiments, R 5a and R 5b One of is H and the other is F or -CH3. In some embodiments, R 5a and R 5b One of them is H and the other is F.

[0226] In some embodiments, R 5a and R 5b are each independently halogen. In some embodiments, R 5a and R 5b are independently chlorine, fluorine, or bromine. 5a and R 5b are independently chlorine or fluorine. 5a and R 5b In some embodiments, R 5a and R 5b In some embodiments, R 5a and R 5b One of them is fluorine.

[0227] In some embodiments, R 5a and R 5b Together with the carbon atom to which it is attached, it forms a C3-C6 cycloalkyl group. 5a and R 5b Together with the carbon atom to which it is attached, it forms a C3-C4 cycloalkyl group. 5a and R 5b Together with the carbon atom to which it is attached, it forms a cyclopropyl group. 5a and R 5b Together with the carbon atom to which it is attached, it forms a cyclobutyl group.

[0228] In some embodiments, R 6 is C1-C6 alkyl, C1-C6 haloalkyl, or C3-C6 cycloalkyl. 6 is C1-C3 alkyl, C1-C3 haloalkyl, or C3-C4 cycloalkyl. 6 It is -CH3, -CHF2, or cyclopropyl.

[0229] In some embodiments, the ring D group moiety is and Y is CR5a R 5b In some embodiments, the ring D group portion is W is O, and Y is CR 5a R 5b In some embodiments, the ring D group portion is W is the key, and Y is the CR 5a R 5b In some embodiments, the ring D group portion is and Y is S. In some embodiments, the ring D group moiety is W is O, and Y is S. In some embodiments, the ring D group moiety is W is a bond, and Y is S. In some embodiments, the ring D group moiety is and Y are bonds.

[0230] In some embodiments, R 6 is C1-C6 alkyl. In some embodiments, R 6 is C1-C3 alkyl. In some embodiments, R 6 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 6 It is -CH3.

[0231] In some embodiments, R 6 is C1-C6 haloalkyl. 6 is a C1-C6 haloalkyl group containing 1 to 13 halogen atoms. 6 is a C1-C6 haloalkyl group containing 1-7 halogen atoms. 6 is C1-C3 haloalkyl. 6 is a C1-C3 haloalkyl group containing 1-7 halogen atoms. 6 is a C1-C2 haloalkyl group containing 1-5 halogen atoms. In some embodiments, the halogen atoms are independently selected from the group consisting of chlorine atoms, bromine atoms, and fluorine atoms. In some embodiments, the halogen atoms are independently selected from the group consisting of chlorine atoms and fluorine atoms. In some embodiments, the halogen atoms are all fluorine atoms. In some embodiments, the halogen atoms are a combination of chlorine atoms and fluorine atoms. In some embodiments, R 6 is -CF3, -CCl3, -CF2Cl, -CFCl2, -CHF2, -CH2F, -CHCl2, -CH2F, or -CHFCl. In some embodiments, R 6 It is -CHF2.

[0232] In some embodiments, R6 is a C3-C6 cycloalkyl group. 6 is a C3-C5 cycloalkyl group. 6 is a C3-C4 cycloalkyl group. 6 is cyclopropyl, cyclobutyl, or cyclopentyl. In some embodiments, R 6 In some embodiments, R 6 It is cyclobutyl.

[0233] In some embodiments, R 7 is H, halogen, C3-C6 cycloalkyl, -NH-(heterocyclyl consisting of 3 to 6 atoms), -NH-(C1-C6 alkyl), -NH-(C3-C6 cycloalkyl), -O-(heterocyclyl consisting of 3 to 6 atoms), -O-(C1-C6 alkyl), or -O-(C3-C6 cycloalkyl). In some embodiments, R 7 is H, halogen, C3-C4 cycloalkyl, -NH-(heterocyclyl consisting of 4 to 5 atoms), -NH-(C1-C3 alkyl), -NH-(C3-C5 cycloalkyl), -O-(C1-C3 alkyl), -O-(heterocyclyl consisting of 4 to 5 atoms), or -O-(C3-C5 cycloalkyl). In some embodiments, R 7 is H, Cl, cyclopropyl, -NH(CH2CH3), -NH(cyclopropyl), -OCH2CH3, -O(cyclopropyl),

[0234] In some embodiments, R 7 It’s H.

[0235] In some embodiments, R 7 is halogen. In some embodiments, R 7 is chlorine, fluorine, or bromine. In some embodiments, R 7 is chlorine or fluorine. In some embodiments, R 7 It's chlorine.

[0236] In some embodiments, R 7 is a C3-C6 cycloalkyl group. 7 is a C3-C4 cycloalkyl group. 7 In some embodiments, R 7 It is cyclopropyl.

[0237] In some embodiments, R 7 is -NH-(3 to 6 atoms consisting of heterocyclic group). In some embodiments, R 7 is -NH-(4 to 5 atoms consisting of heterocyclic group). In some embodiments, R7 The heterocyclyl moiety contains 1-2 heteroatoms independently selected from the group consisting of nitrogen and oxygen. 7 The heterocyclyl group portion of contains 1 oxygen atom. In some embodiments, R 7 The heterocyclyl group portion of contains 1 nitrogen atom. In some embodiments, R 7 The heterocyclyl group portion is selected from the group consisting of: In some embodiments, R 7 yes

[0238] In some embodiments, R 7 is -NH-(C1-C6 alkyl). In some embodiments, R 7 is -NH-(C1-C3 alkyl). In some embodiments, R 7 is -N(H)CH3, -NH(CH2CH3), -NH(CH2CH2CH3), or -N(H)CH(CH3)2. In some embodiments, R 7 It is -NH(CH2CH3).

[0239] In some embodiments, R 7 is -NH-(C3-C6 cycloalkyl). In some embodiments, R 7 is -NH-(C3-C5 cycloalkyl). In some embodiments, R 7 is -NH-(C3-C4 cycloalkyl). In some embodiments, R 7 is -NH-(cyclopropyl), -NH-(cyclobutyl), -NH-(cyclopentyl), or -NH-(cyclohexyl). In some embodiments, R 7 It is -NH-(cyclopropyl).

[0240] In some embodiments, R 7 is -O-(3 to 6 atoms consisting of heterocyclic group). In some embodiments, R 7 is -O-(4 to 5 atoms consisting of heterocyclic group). In some embodiments, R 7 The heterocyclyl moiety contains 1-2 heteroatoms independently selected from the group consisting of nitrogen and oxygen. 7 The heterocyclyl group portion of contains 1 oxygen atom. In some embodiments, R 7 The heterocyclyl group portion of contains 1 nitrogen atom. In some embodiments, R 7 The heterocyclyl group portion is selected from the group consisting of: In some embodiments, R 7 yes

[0241] In some embodiments, R 7 is -O-(C1-C6 alkyl). In some embodiments, R 7 is -O-(C1-C3 alkyl). In some embodiments, R 7 is -OCH3, -OCH2CH3, -OCH2CH2CH3, or -OCH(CH3)2. In some embodiments, R 7 It is -OCH2CH3.

[0242] In some embodiments, R 7 Is -O-(C3-C6 cycloalkyl). In some embodiments, R 7 is -O-(C3-C5 cycloalkyl). In some embodiments, R 7 is -O-(C3-C4 cycloalkyl). In some embodiments, R 7 is -O-(cyclopropyl), -O-(cyclobutyl), -O-(cyclopentyl), or -O-(cyclohexyl). 7 It is -O-(cyclopropyl).

[0243] In some embodiments, R 9 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH. 9 is H, C1-C3 alkyl, C1-C3 haloalkyl, or C1-C3 alkyl-OH. 9 It is H, -CH3, -CF3, or -CH2OH.

[0244] In some embodiments, R 9 It’s H.

[0245] In some embodiments, R 9 is C1-C6 alkyl. In some embodiments, R 9 is C1-C3 alkyl. In some embodiments, R 9 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 9 It is -CH3.

[0246] In some embodiments, R 9 is C1-C6 haloalkyl. 9 is a C1-C6 haloalkyl group containing 1 to 13 halogen atoms. 9 is a C1-C6 haloalkyl group containing 1-7 halogen atoms. 9 is C1-C3 haloalkyl. 9is a C1-C3 haloalkyl group containing 1-7 halogen atoms. 9 is a C1-C2 haloalkyl group containing 1-5 halogen atoms. In some embodiments, the halogen atoms are independently selected from the group consisting of chlorine atoms, bromine atoms, and fluorine atoms. In some embodiments, the halogen atoms are independently selected from the group consisting of chlorine atoms and fluorine atoms. In some embodiments, the halogen atoms are all fluorine atoms. In some embodiments, the halogen atoms are a combination of chlorine atoms and fluorine atoms. In some embodiments, R 9 is -CF3, -CCl3, -CF2Cl, -CFCl2, -CHF2, -CH2F, -CHCl2, -CH2F, or -CHFCl. In some embodiments, R 9 Yes -CF3.

[0247] In some embodiments, R 9 is C1-C6 alkyl-OH. In some embodiments, R 9 is C1-C3 alkyl-OH. In some embodiments, R 9 is -CH2OH, -CH2CH2-OH, -CH2CH2CH2-OH, or -C(CH3)2-OH. In some embodiments, R 9 It is -CH2OH.

[0248] In some embodiments, the compound has a structure represented by Formula (IA) or (IB):

[0249]

[0250] where R 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b 、R 4 、R 6 、R 7 、R 9 、R 10 , X, Y, Z 1 、Z 2 and the ring D group moiety are as described for the compound of formula (I).

[0251] In some embodiments, the compound has a structure represented by Formula (IC), (ID), (IE), or (IF):

[0252]

[0253] where R 1a 、R 1b 、R2a 、R 2b 、R 3a 、R 3b 、R 4 、R 6 、R 7 、R 9 、R 10 , W, X, Y, Z 1 and Z 2 are as defined for the compounds of formula (I).

[0254] In some embodiments, the compound has a structure represented by Formula (IG), (IH), (II), (IJ), (IK), (IL), (IM), or (IN):

[0255]

[0256] where R 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b 、R 4 、R 6 、R 7 、R 9 、R 10 , W, X and Y are as defined for the compound of formula (I).

[0257] In some embodiments, the compound has a structure represented by Formula (II-A), (II-B), (II-C), (II-D), (II-E), (II-F), (II-G), (II-H), (II-I), (II-J), (II-K), or (II-L):

[0258]

[0259]

[0260] where R 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b 、R 4 、R 5a 、R 5b 、R 6 、R 7 、R 8 and R 9 are as defined for compounds of formula (I), and p of formula (II-G) is 1 or 2.

[0261] In some embodiments, the compound has a structure represented by Formula (III-A), (III-B), (III-C), or (III-D):

[0262]

[0263] where R 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b 、R 4 、R 5a 、R 5b 、R 7 and R 9 are as defined for the compounds of formula (I).

[0264] In some embodiments, the compound has a structure shown in Formula (IV-A), (IV-B), or (IV-C):

[0265]

[0266]

[0267] where R 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b 、R 4 、R 5a 、R 5b 、R 6 、R 7 and R 9 are as defined for the compounds of formula (I).

[0268] In some embodiments, the compound has a structure represented by Formula (VA), (VB), (VC), (VD), (VE), (VF), or (VG):

[0269]

[0270]

[0271] where R 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b 、R 4 、R 5a 、R5b 、R 6 、R 7 and R 9 are as defined for the compounds of formula (I).

[0272] In some embodiments, the compound has a structure represented by Formula (VI-A), (VI-B), (VI-C), or (VI-D):

[0273]

[0274] where R 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b 、R 4 、R 5a 、R 5b 、R 6 、R 7 and R 9 are as defined for the compounds of formula (I).

[0275] Table 1. Representative compounds of the present invention

[0276]

[0277]

[0278]

[0279]

[0280]

[0281] In some embodiments, the present invention provides a compound selected from the group consisting of compounds No. 1-45 in Table 1, or a tautomer, a stereoisomer, or a pharmaceutically acceptable salt thereof. In some embodiments, the present invention provides a compound selected from the group consisting of compounds No. 47-52 in Table 1, or a tautomer, a stereoisomer, or a pharmaceutically acceptable salt thereof.

[0282] In one aspect, the present invention provides a method for preparing a compound of formula (I) according to the present invention. In another aspect, the present invention provides an intermediate compound for preparing a compound of formula (I). The present invention also provides a compound as an assay probe and optionally labeled with, for example, a fluorescent marker. In another aspect, the present invention provides a method for determining Cbl-b inhibition. In one variation, the present invention provides a method for determining Cbl-b inhibition, comprising pre-incubating Cbl-b with an assay probe (e.g., an assay probe labeled with a fluorescent marker), then exposing the Cbl-b / assay probe mixture to a candidate compound, and then using, for example, a FRET signal detection method to determine whether the assay probe is displaced by the candidate compound and the extent of the displacement.

[0283] In any pharmaceutical composition, method, or kit described herein, the Cbl-b inhibitor may be selected from one or more Cbl-b inhibitors disclosed in the following patent applications: compounds 1-719 (including "a" and "b" variants thereof) of International Patent Application WO 2019 / 148005, or compounds represented by any of Formula (IA), (I), (II-A), (II), (III-A), (III), or (IV). The contents of International Patent Application WO 2019 / 148005 are incorporated herein by reference in their entirety.

[0284] Representative compounds useful in the pharmaceutical compositions and methods of the present invention are listed in Table 1A.

[0285] Table 1A

[0286]

[0287]

[0288] In some embodiments, provided for use in the compositions and methods of the present invention are compounds selected from Compound Nos. 53-61 in Table 1A, or tautomers, stereoisomers, or pharmaceutically acceptable salts thereof.

[0289] In one embodiment, the small molecule Cbl-b inhibitor used in the methods, pharmaceutical compositions, and kits disclosed herein has a molecular weight of 1,000 Daltons or less, or about 1,000 Daltons or less. In another embodiment, the small molecule Cbl-b inhibitor has a molecular weight of 900 Daltons or less, or about 900 Daltons or less. In another embodiment, the small molecule Cbl-b inhibitor has a molecular weight of 800 Daltons or less, or about 800 Daltons or less. In another embodiment, the small molecule Cbl-b inhibitor has a molecular weight of 750 Daltons or less, or about 750 Daltons or less. In another embodiment, the small molecule Cbl-b inhibitor has a molecular weight of 700 Daltons or less, or about 700 Daltons or less. In another embodiment, the small molecule Cbl-b inhibitor has a molecular weight of 650 Daltons or less, or about 650 Daltons or less. In another embodiment, the small molecule Cbl-b inhibitor has a molecular weight of 600 Daltons or less, or about 600 Daltons or less. For any of the foregoing embodiments, the lower molecular weight limit of the small molecule Cbl-b inhibitor can be 100 Daltons, 200 Daltons, or 300 Daltons, or about 100 Daltons, about 200 Daltons, or about 300 Daltons.

[0290] The following schemes describe methods for synthesizing the compounds disclosed herein. Mixtures of stereoisomers produced during the synthesis, such as racemic mixtures of the final compounds, can be separated into their individual enantiomers using conventional chromatographic methods, such as supercritical fluid chromatography coupled with a chiral stationary phase, chiral column chromatography, or other methods known in the art.

[0291] Plan I

[0292]

[0293] where Z 1 、R 6 and R 10 are as defined for the compounds of formula (I); X′ is Br or NH2; R a and R b are suitable protecting groups; and (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b and R 4 .

[0294] Compounds of formula I-9 can be synthesized as described in Scheme 1. Boronic acid ester I-1 is coupled with olefin I-2 under rhodium catalysis to provide ester I-3. Triazoles are assembled via hydrazide formation (I-4), cyclization, and desulfurization to provide compound I-6. Compound I-6, where X' is NH2, can be reacted with bromoester I-8 to provide compound I-9, which can then be converted to compound I-10 by reductive amination with a substituted piperidine. Compound I-6, where X' is Br, can be reacted with piperidine derivative I-7 under palladium catalysis to directly provide compound I-10.

[0295] Option Ia.

[0296]

[0297] where Z 1 、R 6 、R 9 and R 10 are as defined for the compounds of formula (I); X′ is Br or NH2; R a and R b are suitable protecting groups; and (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b and R 4 .

[0298] Compounds of formula I-9a can be synthesized as outlined in Scheme Ia. Boronic ester I-1 is coupled with olefin I-2 under rhodium catalysis to provide ester I-3. Triazoles are assembled via hydrazide formation (I-4), cyclization, and desulfurization to provide compound I-6. Compound I-6, where X′ is NH2, can be reacted with bromoester I-8a to provide compound I-9a, which is subsequently converted to compound I-10a by reductive amination with a substituted piperidine. Compound I-6, where X′ is Br, can be reacted with piperidine derivative I-7a under palladium catalysis to directly provide compound I-10a.

[0299] Option II.

[0300]

[0301] where R 6 As defined for compounds of formula (I); and (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R 2a 、R 2b 、R3a 、R 3b and R 4 .

[0302] Scheme II outlines the synthesis of compounds of formula II-4. Aldehyde II-1 is converted to the piperidine derivative II-2 by reductive amination with a substituted piperidine followed by ester hydrolysis under alkaline conditions. Piperidine II-2 is then coupled to amine I-6 using a coupling agent such as HATU or T3P to provide compound II-4.

[0303] Option II-a.

[0304]

[0305] where Z 2 、R 6 and R 10 are as defined for the compounds of formula (I); (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b and R 4 ; R b is a suitable protecting group; Y b is an alkyl group (such as methyl), a cycloalkyl group (such as cyclopropyl), a haloalkyl group (such as bromomethyl) or CHO; and Y c It is hydroxyl or NH2.

[0306] Scheme II-a summarizes the synthesis of compounds of formula II-5a. Compounds of formula II-1a can be converted to compounds II-2a by oxidation to aldehydes with reagents such as SeO2, or bromination with Br2, followed by reductive amination or direct displacement with substituted piperidines to form compounds of formula II-3a. Compound II-3a is then subjected to ester hydrolysis under alkaline conditions to provide compounds II-4a (Y c =OH). The acid II-4a is then coupled to the amine I-6 using a coupling agent such as HATU or T3P to provide the amide II-5a.

[0307] Option III.

[0308]

[0309] where Z 1 、R 6 and R 10 are as defined for the compounds of formula (I), (R) n corresponds to R as described for the compound of formula (I)1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b and R 4 .

[0310] Compounds of formula III-4 can be prepared according to Scheme III. Alcohol III-1 is converted to ester III-2 by conversion to a halide, displacement with a cyanide, and then hydrolysis. The triazole is formed as described in Scheme I. The bromide is converted to aniline III-3 by palladium-catalyzed coupling with tert-butyl carbamate, followed by deprotection. Compound III-3 is then purified as described in Scheme I to provide compound III-4.

[0311] Scheme III-a.

[0312]

[0313] where Z 1 、R 6 、R 9 and R 10 are as defined for the compounds of formula (I), and (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b and R 4 .

[0314] Compounds of formula III-4a can be prepared according to Scheme III-a. Alcohol III-1 is converted to ester III-2 by conversion to a halide, displacement with a cyanide, and then hydrolysis. The triazole is formed as described in Scheme Ia. The bromide is converted to aniline III-3 by palladium-catalyzed coupling with tert-butyl carbamate, followed by deprotection. Compound III-3 is then purified as described in Scheme Ia to provide compound III-4a.

[0315] Option IV.

[0316]

[0317] where Z 1 、R 6 and R 10 are as defined for the compounds of formula (I), and (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R2a 、R 2b 、R 3a 、R 3b and R 4 .

[0318] Scheme IV shows a method for preparing compounds of formula IV-5. Ester IV-1 is converted to α-hydroxy ester IV-2 by deprotonation and treatment with an oxidizing agent (such as oxaziridine). The ester is refined to triazole IV-3 according to Scheme I. The hydroxyl group is converted to fluoride IV-4 by DAST. Compound IV-4 is then reacted completely as shown in Scheme I to provide compound IV-5.

[0319] Scheme IV-a.

[0320]

[0321] where Z 1 、R 6 、R 9 and R 10 are as defined for the compounds of formula (I), and (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b and R 4 .

[0322] Scheme IV-a shows a method for preparing a compound of formula IV-5a. Ester IV-1 is converted to α-hydroxy ester IV-2 by deprotonation and treatment with an oxidizing agent (such as oxaziridine). The ester is refined to triazole IV-3 according to Scheme Ia. The hydroxyl group is converted to fluoride IV-4 by DAST. Compound IV-4 is then reacted completely as shown in Scheme Ia to provide compound IV-5a.

[0323] Plan V

[0324]

[0325] where R 6 、R 7 、R 10 and Z 1 are as defined for the compounds of formula (I), and (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3band R 4 .

[0326] The synthesis of compounds of formula V-7 is provided in Scheme V. Trichloropyridine V-1 is coupled with dimethyl malonate to provide diester V-2. Diester V-2 is reacted with allyl bromide and then reduced to produce dihydroxide V-3. Compound V-3 is then reacted under Mitsunobu conditions to provide oxetane V-4. The allyl group is oxidized to an acid and then converted to triazole V-5 as shown in Scheme I. Subsequent reaction of V-5 with nucleophiles such as amines or alcohols, or organometallic reagents under palladium catalysis produces V-6, which is then coupled with indole 1-7 to provide compound V-7.

[0327] Plan Va.

[0328]

[0329] where R 6 、R 7 、R 9 、R 10 and Z 1 are as defined for the compounds of formula (I), and (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b and R 4 .

[0330] The synthesis of compounds of formula V-7a is provided in Scheme Va. Trichloropyridine V-1 is coupled with dimethyl malonate to provide diester V-2. Diester V-2 is reacted with allyl bromide and then reduced to generate dihydroxide V-3. Compound V-3 is then reacted under Mitsunobu reaction conditions to provide oxetane V-4. The allyl group is oxidized to an acid and then converted to triazole V-5 as shown in Scheme Ia. V-5 is subsequently reacted with nucleophiles such as amines or alcohols, or organometallic reagents under palladium catalysis to generate V-6, which is then coupled with indole 1-7a to provide compound V-7a.

[0331] Scheme VI.

[0332]

[0333] where R 10 and Z 1 are as defined for the compounds of formula (I), and (R) n corresponds to R as described for the compound of formula (I)1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b and R 4 .

[0334] Intermediate compounds of formula I-7 employed in the general synthetic method outlined in Scheme I can be synthesized according to Scheme VI. Aldehyde VI-1 is coupled with a substituted piperidine under reductive amination conditions to provide piperidine intermediate VI-2, which is treated with ammonia to provide compounds of formula I-7.

[0335] Plan VI-a.

[0336]

[0337] where R 9 、R 10 and Z 1 are as defined for the compounds of formula (I), and (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b and R 4 .

[0338] The intermediate compounds of formula I-7a employed in the general synthetic method outlined in Scheme 1a can be synthesized according to Scheme VI-a. Aldehyde VI-1a is coupled with a substituted piperidine under reductive amination conditions to provide piperidine intermediate VI-2a, which is treated with ammonia to provide compounds of formula I-7a.

[0339] Scheme VII.

[0340]

[0341] where R 10 and Z 1 are as defined for the compounds of formula (I), and (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b and R 4 .

[0342] Scheme VII outlines the synthesis of compounds of formula VII-5. Bromide VII-1 is coupled with 3-nitrophenylboronic acid to provide nitro compound VII-2. The ester of compound VII-2 is converted to a triazole as shown in Scheme I, and the nitro group is reduced to provide amine VII-3, which is then purified as shown in Scheme I to provide VII-5.

[0343] Scheme VII-a.

[0344]

[0345] where R 9 、R 10 and Z 1 are as defined for the compounds of formula (I), and (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b and R 4 .

[0346] Scheme VII-a outlines the synthesis of compounds of formula VII-5a. Bromide VII-1 is coupled with 3-nitrophenylboronic acid to provide nitro compound VII-2. The ester of compound VII-2 is converted to a triazole as shown in Scheme Ia, and the nitro group is reduced to provide amine VII-3, which is then purified as shown in Scheme Ia to provide VII-5a.

[0347] Scheme VIII

[0348]

[0349] where R 9 、R 10 and Z 1 are as defined for the compounds of formula (I), and (R) n corresponds to R as described for the compound of formula (I) 1a 、R 1b 、R 2a 、R 2b 、R 3a 、R 3b and R 4 , and P is a protecting group, such as pivaloyl.

[0350] Scheme VIII summarizes the synthesis of compounds represented by general formula VIII-6. Hydroxybromide VIII-1 is converted to sulfide (VIII-2) by displacement with a thiol source such as trityl-SH (Croft, Rosemary A. et al. Chemistry-A European Journal, 24(4), 818-821; 2018). Triazole is synthesized by S-termining with triazole chloride. N Ar displacement for installation affords VIII-4, which is then coupled with an isoindolinone (e.g., I-5) to afford VIII-5. The protecting group is then removed under Ni catalysis (Correa, Arkaitz et al., Journal of the American Chemical Society, 136(3), 1062-1069; 2014) to provide analogs VIII-6.

[0351] Compounds 1, 2, 10, 22, 23, 24, 32, 36, 39, 40, 44, and 49-51 exhibited an IC of less than 1 nM in the Cbl-b inhibition assay of Biological Example 1. 50 Compounds 3, 4, 11, 13-20, 25, 26, 28, 31, 33-35, 37, 38, 41-43, 45, 48, and 52 exhibited IC values ​​of 1 nM to 5 nM in the Cbl-b inhibition assay of Biological Example 1. 50 Compounds 5-9, 12, 21, 27, 29, 30, and 47 exhibited IC values ​​of 5.1-100 nM in the Cbl-b inhibition assay of Biological Example 1. 50 Value, and in one embodiment, for use in the pharmaceutical compositions and methods disclosed herein.

[0352] In various embodiments, and as further described herein, the compounds provided herein (as well as compositions comprising the compounds of the present invention, and methods of using the compounds or compositions) have an IC of less than 1 nM, between 1 nM and 5 nM, between 5.1 and 100 nM, or greater than 100 nM as determined by the Cbl-b inhibition assay of Biological Example 1. 50 In further embodiments, and as further described herein, the compounds provided herein (as well as compositions comprising the compounds of the present invention, and methods of using the compounds or compositions) have an IC value of less than 1 nM as measured by the Cbl-b inhibition assay of Biological Example 1. 50In further embodiments, and as further described herein, the compounds provided herein (as well as compositions comprising the compounds of the present invention, and methods of using the compounds or compositions) have an IC between 1 nM and 5 nM as measured by the Cbl-b inhibition assay of Biological Example 1. 50 In further embodiments, and as further described herein, the compounds provided herein (as well as compositions comprising the compounds of the present invention, and methods of using the compounds or compositions) have an IC between 5.1 and 100 nM as measured by the Cbl-b inhibition assay of Biological Example 1. 50 In further embodiments, and as further described herein, the compounds provided herein (as well as compositions comprising the compounds of the present invention, and methods of using the compounds or compositions) have an IC greater than 100 nM as measured by the Cbl-b inhibition assay of Biological Example 1. 50 value.

[0353] For the secretion of IL-2 by immune cells (e.g., T cells) co-stimulated with anti-CD3 antibodies and anti-CD28 antibodies, the compounds provided herein (and compositions comprising the compounds of the present invention) can induce a change of ≤10-fold, between 11-15-fold, or greater than 15-fold compared to baseline at an inhibitor concentration of 1 micromolar or 0.3 micromolar.

[0354] For the secretion of IL-2 by immune cells (e.g., T cells) stimulated with anti-CD3 antibodies, the compounds provided herein (and compositions comprising the compounds of the present invention) can induce a change of ≤0.33-fold, between 0.34-0.66-fold, or greater than 0.66-fold compared to baseline at a 3 micromolar or 1 micromolar inhibitor concentration.

[0355] For CD25 staining on the cell surface of immune cells (e.g., T cells) co-stimulated with anti-CD3 antibodies and anti-CD28 antibodies, the compounds provided herein (and compositions comprising the compounds of the present invention) can induce a change of ≤1.24-fold, between 1.25-1.39-fold, or greater than 1.39-fold compared to baseline at a 1 micromolar or 0.3 micromolar inhibitor concentration.

[0356] For CD25 staining on the cell surface of immune cells (e.g., T cells) stimulated with anti-CD3 antibodies, the compounds provided herein (and compositions comprising the compounds of the present invention) can induce a change of ≤1.04-fold, between 1.05-1.14-fold, or greater than 1.14-fold compared to baseline at a 3 micromolar or 1 micromolar inhibitor concentration.

[0357] III. Methods, Drugs, and Uses

[0358] The present invention provides methods for modulating the activity of immune cells (e.g., T cells, B cells, or NK cells), for example, by contacting the immune cells with an effective amount of a Cbl-b inhibitor of the present invention or a composition thereof. The present invention also provides an in vitro method for generating immune cells having modulated activity (referred to herein as "modified immune cells"), wherein the modified immune cells can be administered to an individual in need (e.g., an individual with cancer) by an ex vivo method. The present invention further provides an in vivo method for modulating a response in an individual in need (e.g., an individual with cancer), wherein the method comprises administering an effective amount of a Cbl-b inhibitor of the present invention or a composition thereof. In addition, the present invention provides an in vitro method for generating an amplified lymphocyte population after lymphatic modulation in an individual, wherein the lymphatic modulation occurs due to administration of an effective amount of a Cbl-b inhibitor of the present invention or a composition thereof to the individual. In addition, the amplified lymphocyte population can then be administered to an individual with cancer. In some embodiments, the modified immune cells or the expanded lymphocyte population are produced from a biological sample comprising immune cells obtained from the individual (e.g., a blood sample comprising peripheral blood mononuclear cells or a tumor biopsy comprising tumor infiltrating lymphocytes (TILs)).

[0359] In addition, the present invention provides Cbl-b inhibitors for use as therapeutically active substances. The present invention provides Cbl-b inhibitors for treating or preventing diseases or conditions associated with Cbl-b activity. In addition, the present invention provides Cbl-b inhibitors for treating cancer. The present invention further provides the use of Cbl-b inhibitors in the preparation of medicaments for treating or preventing diseases or conditions associated with Cbl-b activity. The present invention also provides the use of Cbl-b inhibitors in the preparation of medicaments for treating cancer. In addition, the present invention provides treatment methods, medicaments, and uses comprising Cbl-b inhibitors as part of a combination therapy for treating cancer, wherein the combination therapy for treating cancer involves one or more of an immune checkpoint inhibitor, an anti-tumor agent, and radiotherapy.

[0360] In some embodiments of the methods of treatment, medicaments and uses of the present invention, the cancer is a hematological cancer, such as lymphoma, leukemia, or myeloma. In other embodiments of the methods of treatment, medicaments and uses of the present invention, the cancer is a non-hematological cancer, such as sarcoma, carcinoma, or melanoma.

[0361] Hematological cancers include, but are not limited to, one or more leukemias, such as B-cell acute lymphoblastic leukemia ("BALL"), T-cell acute lymphoblastic leukemia ("TALL"), acute lymphoblastic leukemia (ALL); one or more chronic leukemias, including, but not limited to, chronic myeloid leukemia (CML) and chronic lymphocytic leukemia (CLL); other hematological cancers or hematological diseases, including, but not limited to, B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma (BCL), and leukemia. The term "preleukemia" refers to a variety of hematologic disorders characterized by ineffective production (or dysplasia) of myeloid blood cells, including diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, follicular small cell lymphoma or follicular large cell lymphoma, malignant lymphoproliferative disorders, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndrome, non-Hodgkin lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom macroglobulinemia, and "preleukemias," which are a group of hematologic disorders characterized by ineffective production (or dysplasia) of myeloid blood cells.

[0362] Non-hematological cancers include, but are not limited to, neuroblastoma, renal cell carcinoma, colon cancer, colorectal cancer, breast cancer, epithelial squamous cell carcinoma, melanoma, gastric cancer, brain cancer, lung cancer (e.g., NSCLC), pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, prostate cancer, testicular cancer, thyroid cancer, uterine cancer, adrenal cancer, and head and neck cancer.

[0363] In some cases, the effectiveness of a Cbl-b inhibitor in treating a disease or condition (e.g., cancer) is determined by evaluating clinical outcomes (e.g., reduction in tumor size or number of tumors, and / or survival). In some embodiments, "treating cancer" comprises evaluating a patient's response to a treatment regimen according to the Response Evaluation Criteria in Solid Tumors (RECIST version 1.1), as described in (see, e.g., Eisenhauer et al., Eur J Cancer, 45:228-247, 2009; and Nishino et al., Am J Roentgenol, 195:281-289, 2010). Response criteria for determining an anti-tumor response of interest according to RECIST 1.1 include: complete remission (CR); partial remission (PR); progressive disease (PD); and stable disease (SD).

[0364] A. Cell Isolation and Processing

[0365] The present invention provides methods for preparing and processing immune cells (e.g., modified immune cells) generated and used in the methods of the present invention. As used herein, the term "modified immune cells" refers to immune cells or cell populations comprising immune cells that have been cultured, incubated, and / or contacted with an effective amount of a Cbl-b inhibitor to modulate the activity of the immune cells. In some embodiments, the modified immune cells can be used in immunotherapy, for example, in combination with adoptive immunotherapy methods.

[0366] 1. Sample

[0367] In some embodiments, immune cells to be modified or cell populations comprising the immune cells to be modified are isolated from samples, such as biological samples, for example, samples obtained or derived from individuals (e.g., people). In some embodiments, the individual from which immune cells are isolated is an individual suffering from a specific disease or condition (e.g., cancer) or needs cell therapy or will be administered cell therapy. In some embodiments, the individual is a person who needs a specific therapeutic intervention, such as adoptive cell therapy, in which immune cells are separated, processed and / or modified. Therefore, in some embodiments, the cells separated from the individual are primary cells (e.g., primary human cells). As used in the present invention, the term "primary cells" refers to cells separated directly from mammalian biological fluids or tissues (e.g., human biological fluids or tissues).

[0368] In some embodiments, the immune cells to be modified are hematopoietic cells, pluripotent stem cells, myeloid progenitor cells, lymphoid progenitor cells, T cells, B cells, and / or NK cells. As used in the present invention, the term "hematopoietic cells" includes hematopoietic stem cells and hematopoietic progenitor cells. In some embodiments, the immune cells to be modified are present in a heterogeneous cell group or a composition comprising a heterogeneous cell group. For example, the immune cells to be modified can be hematopoietic cells present in a heterogeneous cell group, and the heterogeneous cell group includes cells such as differentiated cells derived from a tissue or organ. In some embodiments, the immune cells to be modified are present in a homogeneous cell group or a composition comprising a homogeneous cell group. For example, the immune cells to be modified can be hematopoietic cells present in a homogeneous cell group containing only hematopoietic cells. In some embodiments, the immune cells to be modified or the cell group comprising the immune cells to be modified include one or more subpopulations of immune cells. For example, one or more subpopulations of immune cells can be CD4+ cells, CD8+ cells, and subpopulations thereof, such as those defined by function, activation state, maturity, differentiation potential, expansion, localization, persistence capacity, surface marker profile, cytokine secretion profile, and / or degree of differentiation.

[0369] In some embodiments, the biological samples described herein include tissues, body fluids, and other samples taken directly from an individual, as well as samples resulting from one or more processing steps (e.g., separation, centrifugation, genetic engineering (e.g., transduction with a viral vector encoding a recombinant chimeric receptor), washing, and / or incubation). The biological sample can be a sample obtained directly from a biological source or a processed sample. Biological samples include, but are not limited to, body fluids such as blood, plasma, serum, cerebrospinal fluid, synovial fluid, urine, and sweat, tissue and organ samples (e.g., samples from tumor-containing tissues or organs), including processed samples derived therefrom. In some embodiments, the biological sample is a biological fluid sample or a biological tissue sample. In some embodiments, the biological sample is a biological tissue sample. In some cases, the biological sample from which the immune cells are derived or isolated is a blood or blood-derived sample, or a product of apheresis or leukocyte separation.

[0370] Exemplary biological samples include whole blood, peripheral blood mononuclear cells (PBMC), white blood cells, bone marrow, thymus, tissue biopsy, tumor, leukemia, lymphoma, lymph node, gut-associated lymphoid tissue, mucosa-associated lymphoid tissue, spleen, other lymphoid tissue, liver, lung, stomach, intestine, colon, kidney, pancreas, breast, bone, prostate, cervix, testicle, ovary, tonsil, or other organs, and / or cells derived therefrom. In the context of cell therapy (e.g., adoptive cell therapy), biological samples include samples from autologous sources (i.e., obtained or derived from an individual in need of cell therapy) and allogeneic sources (i.e., obtained or derived from an individual or source other than the individual in need of cell therapy).

[0371] In some embodiments, the immune cells to be modified or the cell population comprising the immune cells to be modified are derived from a cell line (e.g., a T cell line, a B cell line, an NK cell line, etc.). In some embodiments, the immune cells to be modified or the cell population comprising the immune cells to be modified are obtained from a xenogeneic source, e.g., from a mouse, a rat, a non-human primate, or a pig.

[0372] 2. Cell Processing and Isolation

[0373] In some embodiments, the separation of immune cells to be modified comprises one or more preparation and / or cell separation steps. The one or more cell separation steps can be a non-affinity-based separation or an affinity-based separation. As an example, a non-affinity-based separation can be a centrifugation of a composition comprising immune cells to be modified. In some embodiments, the non-affinity-based separation method comprises a density-based cell separation method, such as by lysing red blood cells and preparing white blood cells from peripheral blood by Percoll or Ficoll gradient centrifugation. Affinity-based separation methods may comprise contacting a composition comprising immune cells to be modified with antibody-coated beads. Antibody-coated beads encompassed by the present invention include, but are not limited to, magnetic beads coated with antibodies (e.g., magnetic beads sold by Life Technologies, Carlsbad, CA). Sold by Miltenyi Biotec Inc., Auburn, CA Microbeads; or EasySep sold by Stemcell Technologies, Vancouver, BC, Canada TM Direct RapidSpheres TM ), the antibody binds to a marker expressed on the surface of the immune cell to be modified. In some embodiments, specific subpopulations of T cells are isolated by positive or negative selection techniques, such as cells that are positive for one or more surface markers (e.g., CD4+, CD8+, etc.) or otherwise express high levels of one or more surface markers (e.g., CD4+, CD8+, etc.). Positive selection can be based on a technique in which target cells (e.g., immune cells to be modified) are bound to an agent and retained for further use. For example, magnetic beads (e.g., magnetic beads coupled to anti-CD3 antibodies) can be used. CD3 people microbeads) carry out positive selection to CD3+ T cells.Negative selection can be based on the technology of retaining target cells (for example, immune cells to be modified) that are not combined with reagent.For example, negative selection can be used to separate total human primary T cells from peripheral blood mononuclear cells (PBMC), wherein a mixture of antibodies for surface markers CD14, CD15, CD16, CD19, CD34, CD36, CD56, CD123 and CD235a is incubated in a sample comprising PBMC, and then passed through the sample via magnetic beads to remove the cells expressing those surface markers and retain the remaining cells in the sample for subsequent processing. In some embodiments, the immune cell or the cell population comprising the immune cell to be modified is washed, centrifuged and / or incubated in the presence of one or more reagents, for example, to remove unwanted components, enrichment of required components, lysis or removal of cells sensitive to specific reagents. In some embodiments, the immune cell is separated based on one or more characteristics (for example, density, adhesion properties, size, sensitivity, and / or resistance to specific components). The cell separation step does not require 100% enrichment or removal of specific cells. In some embodiments, positive selection or enrichment of a specific type of immune cell (e.g., CD4+ T cells) refers to increasing the number or percentage of such cells. In some embodiments, removing or depleting non-target specific types of cells, such as by negative selection, refers to reducing the number or percentage of such cells.

[0374] In some embodiments, the immune cells or cell populations comprising the immune cells are obtained from the circulating blood of an individual, for example, by apheresis or leukapheresis. In some cases, the sample comprising the immune cells to be modified contains lymphocytes (including T cells, B cells, and NK cells), as well as monocytes, granulocytes, red blood cells, and / or platelets, and in some cases, contains cells other than red blood cells and platelets.

[0375] In some embodiments, blood cells collected from an individual are washed to remove the plasma portion and the cell population comprising the immune cells to be modified is placed in a suitable buffer or culture medium for subsequent processing steps. In some embodiments, the cell population comprising the immune cells to be modified is washed with phosphate-buffered saline. In some embodiments, the washing solution does not contain calcium and / or magnesium. In some cases, the washing step is completed by a semi-automatic "flow-through" centrifuge. In some cases, the washing step is completed by tangential flow filtration. In some embodiments, the immune cells to be modified or the cell population containing the immune cells to be modified are resuspended in a variety of suitable buffers after washing, such as calcium-free and / or magnesium-free phosphate-buffered saline. In some embodiments, the components of the blood cell sample are removed and the immune cells to be modified or the cell population comprising the immune cells to be modified are directly resuspended in a suitable cell culture medium.

[0376] Representative methods for processing and / or separating immune cells (e.g., hematopoietic cells) from samples are described in Biological Examples 2 and 3 of the present invention, wherein the sample comprises a cell population (e.g., a sample comprising PBMC) containing the hematopoietic cells. Methods and techniques for processing and / or separating immune cells (e.g., hematopoietic cells, pluripotent stem cells, myeloid progenitor cells, lymphoid progenitor cells, T cells, B cells, and / or NK cells) are well known in the art. See, for example, U.S. Patent Application No. 2017 / 0037369; U.S. Patent Application No. 2012 / 0148553; U.S. Patent No. 6,461,645; U.S. Patent No. 6,352,694; and U.S. Patent No. 7,776,562.

[0377] 3. Incubation and Treatment

[0378] The present invention provides methods for modulating immune cell activity by contacting the immune cells (such as the treated and / or isolated immune cells described above) with an effective amount of a Cbl-b inhibitor described herein. The present invention also provides modified immune cells generated by any of the methods described herein, for example, by culturing a cell population containing immune cells (e.g., the treated and / or isolated immune cells described above) in the presence of an effective amount of a Cbl-b inhibitor to modulate immune cell activity, thereby generating the modified immune cells.

[0379] In some embodiments, the immune cells to be modified (e.g., the treated and / or isolated immune cells described above) are incubated and / or cultured in a suitable culture medium before contacting the immune cells with the Cbl-b inhibitor provided herein. In some embodiments, the immune cells to be modified are incubated and / or cultured in a suitable culture medium to simultaneously contact the immune cells with the Cbl-b inhibitor provided herein.

[0380] The treated and / or separated immune cells to be modified or the cell population comprising the immune cells to be modified can be differentiated and / or amplified in vitro. In some embodiments, the immune cells to be modified are hematopoietic cells, pluripotent stem cells, myeloid progenitor cells, lymphoid progenitor cells, T cells, B cells and / or NK cells. In some embodiments, the immune cells to be modified are incubated in a suitable cell culture medium comprising the Cbl-b inhibitor of the present invention before the immune cells are differentiated and / or amplified. In some embodiments, the immune cells to be modified are incubated in a suitable cell culture medium comprising the Cbl-b inhibitor of the present invention after the immune cells are differentiated and / or amplified. The immune cells are modified (i.e., modified immune cells) when contacted with an effective amount of the Cbl-b inhibitor provided by the present invention to regulate the activity of the immune cells. In some embodiments, the immune cells to be modified are not differentiated and / or amplified in vitro, and therefore are the same cell type as the modified immune cells that have been contacted with the Cbl-b inhibitor. For example, T cells can be cultured in a suitable culture medium comprising a Cbl-b inhibitor without differentiating the T cells. In other embodiments, the immune cells to be modified are differentiated and / or expanded in vitro, and thus are of a different cell type than the modified immune cells that have been contacted with the Cbl-b inhibitor. For example, hematopoietic cells can be incubated in a suitable culture medium containing a Cbl-b inhibitor and other agents that drive the hematopoietic cells to differentiate into mature hematopoietic cells. Therefore, in some cases of the embodiments of the present invention, the modified immune cells are hematopoietic cells, pluripotent stem cells, myeloid progenitor cells, lymphoid progenitor cells, T cells, B cells, and / or NK cells. Methods for expanding and / or differentiating immune cells are well known in the art. See, for example, International Patent Application No. WO 2017 / 037083.

[0381] An effective amount of a Cbl-b inhibitor is an amount or concentration of a Cbl-b inhibitor sufficient to modulate immune cell activity compared to a reference sample. A reference sample can be an immune cell that has not been contacted with a Cbl-b inhibitor. In some embodiments, the concentration of the Cbl-b inhibitor added to a composition (e.g., cell culture medium) comprising the immune cells to be modified is from about 1 pM to about 100 μM, about 5 pM to about 100 μM, about 10 pM to about 100 μM, about 20 pM to about 100 μM, about 40 pM to about 100 μM, about 60 pM to about 100 μM, about 80 pM to about 100 μM, about 1 nM to about 100 μM, about 3 nM to about 100 μM, about 10 nM to about 100 μM, about 15 nM to about 100 μM, about 20 nM to about 100 μM, about 100 nM to about 100 μM, about 150 nM to about 100 μM, about 20 ... 0 μM, about 40 nM to about 100 μM, about 60 nM to about 100 μM, about 80 nM to about 100 μM, about 0.1 μM to about 100 μM, about 0.1 μM to about 90 μM, about 0.1 μM to about 80 μM, about 0.1 μM to about 70 μM, about 0.1 μM to about 60 μM, about 0.1 μM to about 50 μM, about 0.1 μM to about 40 μM, about 0.1 μM to about 30 μM, about 0.1 μM to about 20 μM, about 0.1 μM to about 10 μM, about 0.2 μM to about 10 μM, or about 0.3 μM to about 8 μM. In some embodiments, the concentration of the Cbl-b inhibitor added to the composition (e.g., cell culture medium) containing the immune cells to be modified is about 1 pM, about 2 pM, about 3 pM, about 4 pM, about 5 pM, about 10 pM, about 20 pM, about 30 pM, about 40 pM, about 50 pM, about 60 pM, about 70 pM, about 80 pM, about 90 pM, about 1 nM, about 3 nM, about 5 nM, about 1 In some embodiments, the concentration of the Cbl-b inhibitor added to the composition (e.g., cell culture medium) comprising the immune cells to be modified is about 0.3 μM, about 1 μM, or about 4 μM. In some embodiments, the concentration of the Cbl-b inhibitor added to the composition (e.g., cell culture medium) comprising the immune cells to be modified is about 1 μM or about 8 μM.

[0382] An effective amount of a Cbl-b inhibitor is contacted with the immune cells for a sufficient period of time to modulate the activity of the immune cells compared to a reference sample. A reference sample can be immune cells that have not been contacted with the Cbl-b inhibitor but have been incubated with a composition (e.g., cell culture medium) comprising the immune cells and the Cbl-b inhibitor for the same length of time. In some embodiments, the Cbl-b inhibitor is contacted and / or incubated with the immune cells for about 1 minute to about 1 hour, about 5 minutes to about 1 hour, about 10 minutes to about 1 hour, about 15 minutes to about 1 hour, about 20 minutes to about 1 hour, about 30 minutes to about 1 hour, about 45 minutes to about 1 hour, about 1 hour to about 2 hours, about 1 hour to about 4 hours, about 1 hour to about 6 hours, about 1 hour to about 8 hours, about 1 hour to about 12 hours, about 1 hour to about 24 hours, about 2 hours to about 24 hours, about 6 hours to about 7 hours, about 6 hours to about 24 hours, about 8 hours to about 24 hours, about 10 hours to about 24 hours, about 15 hours to about 24 hours, about 20 hours to about 24 hours, about 12 hours to about 48 hours, about 24 hours to about 48 hours, or about 36 hours to about 48 hours. In some embodiments, the Cbl-b inhibitor is contacted with and / or incubated with the immune cells for about 1 minute, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 30 minutes, about 40 minutes, about 50 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, or about 24 hours. In some embodiments, the Cbl-b inhibitor is contacted with and / or incubated with the immune cells for about 1 day to about 7 days, about 2 days to about 7 days, about 3 days to about 7 days, about 4 days to about 7 days, about 5 days to about 7 days, or about 6 days to about 7 days. In some embodiments, the Cbl-b inhibitor is contacted with and / or incubated with the immune cells for about 7 days to about 14 days, about 14 days to about 21 days, or about 21 days to about 28 days. In some embodiments, the Cbl-b inhibitor is contacted and / or incubated with the immune cells for about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, or about 14 days.

[0383] In some embodiments, the immune cells or cell populations comprising the immune cells are incubated under suitable conditions to induce proliferation, expansion, activation, and / or survival of the immune cells. Suitable conditions during incubation include, but are not limited to, the use of cell culture medium, temperature, incubation time, the presence of stimulants (e.g., anti-CD3 and / or anti-CD28 antibodies), and the presence of any other beneficial agents (e.g., growth factors, cytokines, chemokines, and / or recombinant soluble receptors).

[0384] In some embodiments, suitable conditions for inducing immune cell proliferation, expansion, activation and / or survival include providing stimulation conditions comprising agents capable of activating the immune cells (e.g., NK cells). For example, suitable conditions for inducing T cell proliferation, expansion, activation and / or survival include providing stimulation conditions comprising agents capable of activating intracellular signaling in T cells. The complete activation of T cells generally requires T cell receptor recognition of antigens (referred to herein as "TCR" (signal 1)) and identification of costimulators such as CD28 (signal 2). In some cases, one or more agents turn on or initiate the intracellular signal cascade mediated by the TCR complex in T cells. For example, a first agent can bind to a component of the TCR complex to activate T cells, and a second agent can bind to costimulatory molecules on the surface of T cells to stimulate the activated T cells. In some embodiments, the first agent stimulates TCR / CD3 complex-related signals (e.g., anti-CD3 antibodies) in T cells by specifically binding to CD3. In a further embodiment, the co-stimulatory molecule on the T cell surface can be CD28, and the second agent specifically binds to CD28 (for example, anti-CD28 antibodies). Such agents include but are not limited to antibodies, bivalent antibody fragments and binding molecules, such as those that are specific to TCR complex components (for example, anti-CD3 antibodies) and / or those that are specific to co-stimulatory receptors (for example, anti-CD28 antibodies). In some embodiments, the agent specifically bound to CD3 is an anti-CD3 antibody, a bivalent antibody fragment of an anti-CD3 antibody (for example, (Fab) 2' fragment or a bivalent scFv fragment), a monovalent antibody fragment of an anti-CD3 antibody (for example, a Fab fragment, an Fv fragment or an scFv fragment) or a CD3 binding molecule (for example, an aptamer). In some embodiments, the agent that specifically binds to CD28 is an anti-CD28 antibody, a bivalent antibody fragment of an anti-CD28 antibody (e.g., a (Fab) 2′ fragment or a bivalent scFv fragment), a monovalent antibody fragment of an anti-CD28 antibody (e.g., a Fab fragment, an Fv fragment, or an scFv fragment), or a CD28 binding molecule (e.g., an aptamer). For example, one or more reagents provided herein (e.g., anti-CD3 antibodies and anti-CD28 antibodies) can be bound to a solid support (e.g., beads) or cross-linked with an anti-Fc antibody. In some embodiments, the amplification method step may further comprise the step of adding anti-CD3 antibodies and / or anti-CD28 antibodies to the culture medium. In some embodiments, the stimulator added to the cell culture medium includes one or more cytokines, such as, but not limited to, one or more of IL-2, IL-7, IL-15, and IL-21. For example, IL-2 can be added to the cell culture medium containing the immune cells and the agent (e.g., anti-CD3 antibodies and / or anti-CD28 antibodies) at a concentration of at least about 10 units / mL.

[0385] In some embodiments, suitable conditions for inducing T cell proliferation, amplification, activation and / or survival include providing stimulation conditions or agents capable of activating intracellular signaling by T cell receptor (TCR) complex, and Cbl-b inhibitors as described herein. In some embodiments, the immune cell or the cell group comprising the immune cell are incubated together with a first agent (e.g., anti-CD3 antibodies) that stimulates TCR / CD3 complex-related signals in T cells by specifically binding CD3. In a further embodiment, the immune cell or the cell group comprising the immune cell are incubated together with a first agent (e.g., anti-CD3 antibodies) that stimulates TCR / CD3 complex-related signals in T cells by specifically binding CD3, a second agent (e.g., anti-CD28 antibodies) that is bound to costimulatory molecule CD28, and a Cbl-b inhibitor having a concentration of about 1 pM to about 100 μM (e.g., about 0.3 μM, about 1 μM, or about 4 μM). In some embodiments, suitable conditions for inducing T cell proliferation, expansion, activation and / or survival when a Cbl-b inhibitor is present do not require stimulation by costimulatory molecules (e.g., CD28). Contacting T cells with a Cbl-b inhibitor or a composition thereof can bypass the need for costimulation required for T cells to enter an activated state. In certain embodiments, the immune cells or a cell population comprising the immune cells are incubated with a first agent (e.g., an anti-CD3 antibody) that stimulates TCR / CD3 complex-related signals in T cells by specifically binding to CD3, and with a concentration of about 0.001 μM to about 1,000 μM, about 0.01 μM to about 100 μM, about 0.1 μM to about 10 μM, or about 0.1 μM to about 50 μM (e.g., about 1 μM or about 8 μM).

[0386] In some embodiments of the method for regulating immune cell activity, the immune cell is a T cell, and regulating T cell activity includes increased T cell activation and / or increased T cell proliferation. Even in the presence of an activator (e.g., anti-CD3 antibody) combined with a component of the TCR complex, and in the presence of a stimulator (e.g., anti-CD28 antibody) in conjunction with a costimulatory molecule, the T cells encompassed by the embodiments of the present invention may also be in a tolerant state. In some embodiments, the method for regulating T cell activity is included in the presence of a combination of anti-CD3 antibodies and anti-CD28 antibodies, so that T cells are contacted with an effective amount of a Cbl-b inhibitor or its composition. In some embodiments, the method for regulating T cell activity includes contacting T cells with an effective amount of a Cbl-b inhibitor or its composition, wherein the T cells have previously been contacted with a combination of anti-CD3 antibodies and anti-CD28 antibodies. In some embodiments, regulating T cell activity (e.g., increasing T cell activation and / or increasing T cell proliferation) does not require stimulation via costimulatory CD28 molecules. In some embodiments, the method for regulating T cell activity is included in contacting T cells with an effective amount of a Cbl-b inhibitor or its composition in the presence of anti-CD3 antibodies alone. In some embodiments, the method of modulating T cell activity comprises contacting a T cell with an effective amount of a Cbl-b inhibitor or a composition thereof, wherein the T cell has previously been contacted with one or more agents that activate T cells (e.g., anti-CD3 antibodies), wherein the agents do not include an agent that stimulates a CD28 co-stimulatory molecule (e.g., an anti-CD28 antibody).

[0387] In some embodiments, the immune cell is a T cell, and regulating T cell activity includes increased T cell activation and / or increased T cell proliferation. For example, such as when there is an agent (e.g., anti-CD3 antibody) that activates T cells, and in some further embodiments, when there is an agent (e.g., anti-CD28 antibody) that stimulates T cells, the T cells encompassed by the embodiments of the present invention may be in an activated state. Contacting T cells with a Cbl-b inhibitor or its composition can reduce the threshold required for activation, so in the presence of an activator (e.g., anti-CD3 antibody) and in some further embodiments, there is a stimulating agent (e.g., anti-CD28 antibody), T cell activation and / or proliferation can be increased. In some embodiments, the method for regulating T cell activity is included in the presence of a combination of an anti-CD3 antibody and an anti-CD28 antibody, contacting T cells with an effective amount of a Cbl-b inhibitor or its composition. In some embodiments, the method for regulating T cell activity includes contacting T cells with an effective amount of a Cbl-b inhibitor or its composition, wherein the T cells have previously been contacted with a combination of an anti-CD3 antibody and an anti-CD28 antibody. In some embodiments, regulating T cell activity (e.g., increasing T cell activation and / or increasing T cell proliferation) does not require stimulation by co-stimulatory CD28 molecules. In some embodiments, the method for regulating T cell activity is included in the presence of anti-CD3 antibodies alone, contacting T cells with an effective amount of a Cbl-b inhibitor or a composition thereof. In some embodiments, the method for regulating T cell activity includes contacting T cells with an effective amount of a Cbl-b inhibitor or a composition thereof, wherein the T cells have previously been contacted with one or more agents (e.g., anti-CD3 antibodies) that activate T cells.

[0388] In some embodiments, the immune cells are T cells, and regulating T cell activity comprises reduced T cell dysfunction, including reduced T cell exhaustion, reduced T cell tolerance, and / or reduced T cell incapacity. The general principles of T cell dysfunction are well known in the art (see, for example, Schietinger et al., Trends Immunol., 35: 51-60, 2014). Immune tolerance is a process that is part of the normal function of the immune system. Antigen-specific immune tolerance is characterized by reduced reactivity to antigens due to previous exposure to the antigen. When specific lymphocytes (such as T cells) encounter antigens, lymphocytes may be activated, resulting in an antigen-specific immune response, or lymphocytes (such as T cells) may be inactivated or eliminated, thereby resulting in antigen-specific immune tolerance. In some cases, tolerance can be caused by clonal incapacity, peripheral clonal deletion, T cell suppression, and / or other forms of antigen-specific tolerance. In some embodiments, tolerance can be produced by induction of incapacity or characterized by induction of incapacity. In some cases, disability can be caused by exposing T cells to antigens in the absence of costimulation. In the absence of costimulatory signals, prolonged antigen recognition by TCR alone may lead to disability (i.e., functional anergy). Disabled T cells may be resistant to subsequent antigen attacks and may be able to suppress other immune responses. Typically, in the natural environment, tolerance involves non-reactive or non-generative reactions to autoantigens. However, in some cases, tolerance to "non-self" antigens can be induced. Therefore, in some cases, the same mechanism by which mature T cells that recognize autoantigens in peripheral tissues are unable to subsequently respond to these antigens may also regulate the anergy to foreign or "non-self" antigens (e.g., antigens expressed by cancer cells). Therefore, even in the presence of a stimulatory agent (e.g., an agent combined with a costimulatory molecule such as CD28), the T cells encompassed by the embodiments of the present invention may also be in a tolerant state. Contacting T cells with a Cbl-b inhibitor or its composition provided by the present invention can bypass certain aspects of T cell dysfunction (e.g., T cell tolerance, T cell anergy, and / or T cell exhaustion). In some embodiments, the method of modulating T cell activity (e.g., reducing T cell tolerance, reducing T cell anemia, and / or reducing T cell exhaustion) comprises contacting T cells with an effective amount of a Cbl-b inhibitor or a composition thereof. In some embodiments of the methods of the present invention, modulating T cell activity (e.g., reducing T cell tolerance, reducing T cell anemia, and / or reducing T cell exhaustion) comprises contacting T cells with an effective amount of a Cbl-b inhibitor or a composition thereof in the presence of a combination of an anti-CD3 antibody and an anti-CD28 antibody.In some embodiments of the present invention, the method for regulating T cell activity (for example, reducing T cell tolerance, reducing T cell disability and / or reducing T cell exhaustion) includes contacting T cells with an effective amount of Cbl-b inhibitor or its composition, wherein the T cells have previously been contacted with a combination of anti-CD3 antibodies and anti-CD28 antibodies. In some embodiments, regulating T cell activity (for example, reducing T cell tolerance, reducing T cell disability and / or reducing T cell exhaustion) does not require stimulation via costimulatory CD28 molecules. In some embodiments of the present invention, the method for regulating T cell activity (for example, reducing T cell tolerance, reducing T cell disability and / or reducing T cell exhaustion) includes contacting T cells with an effective amount of Cbl-b inhibitor or its composition in the presence of a single anti-CD3 antibody. In some embodiments, regulating T cell activity (for example, reducing T cell tolerance, reducing T cell disability and / or reducing T cell exhaustion) includes contacting T cells with an effective amount of Cbl-b inhibitor or its composition, wherein the T cells have previously been contacted with one or more agents (for example, single anti-CD3 antibodies) that activate T cells.

[0389] T cell activation and T cell tolerance are both strict control processes for regulating immune response. Therefore, the invention provides a method for regulating T cell activity, wherein regulating T cell activity comprises the T cell activation of increase, the T cell proliferation of increase, the T cell exhaustion of reduction and / or the T cell tolerance of reduction. In some embodiments, the method for regulating T cell activity (for example, the T cell activation of increase, the T cell proliferation of increase, the T cell exhaustion of reduction and / or the T cell tolerance of reduction) comprises contacting T cells with the Cbl-b inhibitor or its composition of effective dose. In some embodiments of the inventive method, regulating T cell activity (for example, increasing T cell activation, increasing T cell proliferation, reducing T cell exhaustion and / or reducing T cell tolerance) comprises contacting T cells with the Cbl-b inhibitor or its composition of effective dose in the presence of a combination of anti-CD3 antibodies and anti-CD28 antibodies. In some embodiments of the present invention, regulating T cell activity (e.g., increasing T cell activation, increasing T cell proliferation, reducing T cell exhaustion, and / or reducing T cell tolerance) includes contacting T cells with an effective amount of a Cbl-b inhibitor or its composition, wherein the T cells have previously been contacted with a combination of anti-CD3 antibodies and anti-CD28 antibodies. In some embodiments, regulating T cell activity (e.g., increasing T cell activation, increasing T cell proliferation, reducing T cell exhaustion, and / or reducing T cell tolerance) does not require stimulation by costimulatory CD28 molecules. In some embodiments of the present invention, regulating T cell activity (e.g., increasing T cell activation, increasing T cell proliferation, reducing T cell exhaustion, and / or reducing T cell tolerance) methods include contacting T cells with an effective amount of a Cbl-b inhibitor or its composition provided by the present invention in the presence of a single anti-CD3 antibody. In some embodiments, a method of modulating T cell activity (e.g., increasing T cell activation, increasing T cell proliferation, decreasing T cell exhaustion, and / or decreasing T cell tolerance) comprises contacting T cells with an effective amount of a Cbl-b inhibitor or a composition thereof, wherein the T cells have previously been contacted with one or more agents that activate T cells (e.g., anti-CD3 antibodies).

[0390] In some embodiments of the present invention, the increased T cell activation includes increasing the generation of one or more cytokines from the T cells or peripheral immune cells (e.g., myeloid cells) in the activated T cell microenvironment. In some embodiments, the one or more cytokines include but are not limited to: IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-13, IL-18, TNFα and GM-CSF. In some embodiments, the cytokine is one or more of the following: IL-2, IFN-γ, TNFα and GM-CSF. In some embodiments, the cytokine is a chemokine. In some embodiments, the one or more chemokines include but are not limited to: IP-10, eotaxin (Eotaxin), GROα, RANTES, MIP-1α, MIP-1β, MIP-2, MCP-1 and MCP-3. Cytokine expression increase can be measured by ELISA.

[0391] In some embodiments of the methods of the present invention, increased T cell activation comprises increasing cell surface expression of one or more T cell activation markers. In some embodiments, the one or more T cell activation markers include, but are not limited to, CD25, CD44, CD62L, CD69, CD152 (CTLA4), CD154, CD137, and CD279. In some embodiments, the T cell activation markers are one or more of the following: CD25, CD69, and CTLA4. Increased expression of cell surface markers can be measured by FACS.

[0392] Methods for experimentally determining increased T cell activation, increased T cell proliferation, decreased T cell exhaustion, and / or decreased T cell tolerance are well known in the art. In some embodiments, representative methods for determining T cell activation can be found in Biological Example 2 provided herein. In some embodiments, representative in vitro and in vivo methods for determining increased T cell activation, increased T cell proliferation, decreased T cell exhaustion, and / or decreased T cell tolerance can be found in Biological Example 3 provided herein.

[0393] In some embodiments of the method for regulating immune cell activity, immune cell is a B cell, and regulating B cell activity comprises increased B cell activation. In some embodiments, increased B cell activation comprises increasing the cell surface expression of one or more B cell activation markers. In some embodiments, the one or more B cell activation markers include but are not limited to: CD69, CD86 and MHC II class (e.g., HLA-DR). In some embodiments, the B cell activation marker is CD69. The expression of cell surface markers increases and can be measured by FACS. In some embodiments, increased B cell activation comprises increasing the activation of proteins in signaling pathways (e.g., those mediated by ERK, JNK and Syk). The increase activation of the protein can be detected by using reagents (e.g., anti-phosphorylation antibodies available in the art) to measure the phosphorylation level on the protein.

[0394] In some embodiments of the method for regulating immune cell activity, the immune cell is a NK cell, and regulating NK cell activity comprises increased NK cell activation. In some embodiments, the increased NK cell activation comprises the secretion of one or more cytokines. In some embodiments, the one or more cytokines include but are not limited to: IFN-γ, TNFα and MIP-1β. The increased cytokine expression can be measured by ELISA. In some embodiments, the increased NK cell activation comprises increasing the cell surface expression of one or more NK cell activation markers. In some embodiments, the one or more NK cell activation markers include but are not limited to: CD69 and CD107a. The increased expression of cell surface markers can be measured by FACS. In some embodiments, the increased NK cell activation includes increasing the killing of target cells (e.g., tumor cells, including primary tumor cells, and cell line-derived tumor cells, such as K562 cell lines).

[0395] Methods for experimentally determining increased B cell activation and increased NK cell activation are well known in the art (see, e.g., Fauriat et al., Blood. 115: 2167-76, 2010; Beano et al., J. Transl. Med., 6: 25 2008; Claus et al., J. Immunol. Methods, 341: 154-64, 2009; and Fujisaki et al., Cancer Res. 69: 4010-4017, 2009). In some embodiments, representative methods for determining B cell activation can be found in Biological Example 3 provided herein. In some embodiments, representative methods for determining NK cell activation can be found in Biological Example 3 provided herein.

[0396] The activity regulation of immune cells (e.g., T cells, B cells or NK cells) can be measured by measuring the baseline value of the target parameter (e.g., cytokine secretion). For example, T cell activation, such as T cell activation in a sample obtained from an in vitro experiment of cells contacted with a Cbl-b inhibitor, can be measured before contacting or applying the Cbl-b inhibitor to determine a baseline value. A reference value for T cell activation is then obtained after contacting or applying the Cbl-b inhibitor. The reference value is compared with the baseline value to determine the amount of T cell activation caused by contacting or applying the Cbl-b inhibitor or its composition. For example, in some embodiments, compared to the baseline value, the immune cell (e.g., T cell) activation in the sample increases by at least 0.1 times, wherein the baseline value is obtained before the immune cell (e.g., T cell) is contacted with the Cbl-b inhibitor or its composition. In some embodiments, immune cell (for example, T cell) activation increases by at least about 0.1 times, about 0.2 times, about 0.3 times, about 0.4 times, about 0.5 times, about 0.6 times, about 0.7 times, about 0.8 times, about 0.9 times, about 1 times, about 2 times, about 4 times, about 6 times, about 8 times, about 10 times, about 20 times, about 30 times, about 40 times, about 50 times, about 75 times or about 100 times (for example, about 0.1 times to about 100 times, or about 1 times to about 100 times) than baseline value. Immune cell activation can be assessed by measuring activated biomarkers such as cytokine secretion increase, activation markers (for example, cell surface markers) cell surface expression increase or downstream signaling pathway phosphorylation increase of protein. The multiple relative to baseline value indicating immune cell activation can be determined for the conditions where the parameters tested and immune cells are processed. For example, to measure T cell activation, a baseline value can be obtained by stimulating T cells with a combination of an anti-CD3 antibody and an anti-CD28 antibody, wherein the cells are not incubated with a Cbl-b inhibitor. A reference value is then obtained by stimulating T cells with a combination of an anti-CD3 antibody and an anti-CD28 antibody, wherein the T cells have been contacted with or are in contact with a Cbl-b inhibitor. A positive response to immune cell activation can then be determined using the reference value obtained. Similar reference value measurements can be obtained and compared with baseline values ​​for assessing T cell activation, T cell proliferation, T cell exhaustion, T cell tolerance, B cell activation, and / or NK cell activation. The measured values ​​of these parameters can be obtained using techniques well known in the art and the techniques provided in Biological Examples 2 and 3.

[0397] As used herein, the term "baseline" or "baseline value" may refer to a measured value or representation before administering a therapeutic agent disclosed herein (e.g., a composition comprising a Cbl-b inhibitor described herein) or at the start of administration of the therapeutic agent. The baseline value can be compared to a reference value to determine an increase or decrease in immune cell function (e.g., increased T cell activation, increased T cell proliferation, decreased T cell exhaustion, and / or decreased T cell tolerance). As used herein, the term "reference" or "reference value" may refer to a measured value or representation after administering a therapeutic agent disclosed herein (e.g., a composition comprising a Cbl-b inhibitor described herein). The reference value can be measured one or more times during the experimental time course, dosage regimen, or treatment cycle or at the completion of the experimental time course, dosage regimen, or treatment cycle. A "reference value" can be an absolute value, a relative value, a value with an upper and / or lower limit, a range of values, an average value, a median value, an average value, or a value compared to a baseline value. Similarly, a "baseline value" can be an absolute value, a relative value, a value with an upper and / or lower limit, a range of values, an average value, a median value, an average value, or a value compared to a reference value. Reference values ​​and / or baseline values ​​can be obtained from one sample (e.g., one sample obtained from an individual), two different samples (e.g., samples obtained from two different individuals), or a group of samples (e.g., samples obtained from a group of two, three, four, five or more individuals).

[0398] In some embodiments, a positive response to T cell activation as measured by cytokine secretion (e.g., IL-2 secretion) of T cells stimulated with a combination of anti-CD3 and anti-CD28 antibodies in the presence of a Cbl-b inhibitor is at least 2.5 times the baseline value of cytokine secretion (e.g., IL-2 secretion) obtained from T cells stimulated with a combination of anti-CD3 and anti-CD28 antibodies in the absence of a Cbl-b inhibitor. In some embodiments, a positive response to T cell activation as measured by surface marker expression (e.g., CD25 surface marker expression) of T cells stimulated with a combination of anti-CD3 and anti-CD28 antibodies in the presence of a Cbl-b inhibitor is at least 1.3 times the baseline value of surface marker expression (e.g., CD25 surface marker expression) obtained from T cells stimulated with a combination of anti-CD3 and anti-CD28 antibodies in the absence of a Cbl-b inhibitor. In some embodiments, a baseline value can be obtained from T cells stimulated with an anti-CD3 antibody alone, wherein the cells are not incubated with a Cbl-b inhibitor. In some embodiments, a positive response to T cell activation as measured by cytokine secretion (e.g., IL-2 secretion) from T cells stimulated with an anti-CD3 antibody alone in the presence of a Cbl-b inhibitor is at least 0.1 times the baseline value of cytokine secretion (e.g., IL-2 secretion) obtained from T cells stimulated with an anti-CD3 antibody alone in the absence of a Cbl-b inhibitor. In some embodiments, a positive response to T cell activation as measured by surface marker expression (e.g., CD25 surface marker expression) from T cells stimulated with an anti-CD3 antibody alone in the presence of a Cbl-b inhibitor is at least 0.6 times the baseline value of surface marker expression (e.g., CD25 surface marker expression) obtained from T cells stimulated with an anti-CD3 antibody alone in the absence of a Cbl-b inhibitor.

[0399] In some cases, the present invention provides a method for generating modified immune cells, comprising culturing a cell population containing immune cells in the presence of an effective amount of a Cbl-b inhibitor provided by the present invention or a composition thereof to modulate immune cell activity, thereby generating the modified immune cells. In some embodiments, the immune cells are T cells, B cells, or natural killer (NK) cells.

[0400] In some embodiments of the method for generating modified immune cells, the immune cells to be modified are cells selected from the group consisting of: hematopoietic cells, pluripotent stem cells, myeloid progenitor cells, lymphoid progenitor cells, T cells, B cells, and NK cells. In some embodiments, the method further comprises culturing the immune cells with stimulants such as cytokines or antibodies (e.g., anti-CD3 antibodies and / or anti-CD28 antibodies) that bind to activation proteins expressed by immune cells. In some embodiments, the immune cells to be modified are in a cell population comprising the immune cells, wherein the cell population is obtained as a sample from an individual. In some embodiments, the immune cells to be modified are in a cell population comprising the immune cells, wherein the cell population is obtained by culturing a biological sample (e.g., a blood sample, a bone marrow sample, etc.) from an individual. In some embodiments, the immune cells are modified by contacting a cell population comprising immune cells with a Cbl-b inhibitor or a composition thereof, thereby generating modified immune cells. In some embodiments, the modified immune cell is a cell selected from the group consisting of: hematopoietic cells, pluripotent stem cells, myeloid progenitor cells, lymphoid progenitor cells, T cells, B cells, and NK cells. In some embodiments, the immune cell is the same cell type as the modified immune cell. For example, the immune cell can be an inactivated T cell, and the modified immune cell can be an activated T cell. In some embodiments, the immune cell is a cell type different from the modified immune cell. For example, the immune cell can be a hematopoietic stem cell, and the modified immune cell can be an NK cell differentiated from the hematopoietic stem cell. In some embodiments of the method for generating modified immune cells, the method further comprises reclaiming the modified immune cell. In some embodiments, the cell population comprising the immune cell, the immune cell, or the modified immune cell are all from an individual (e.g., human). In some embodiments, the immune cell or the modified immune cell are human immune cells or human modified immune cells, respectively.

[0401] The present invention further provides modified immune cells produced by any of the methods described herein, for example, by culturing a cell population comprising immune cells in the presence of an effective amount of a Cbl-b inhibitor to modulate immune cell activity, thereby producing the modified immune cells.

[0402] In some embodiments, the Cbl-b inhibitors provided herein are cell membrane permeable. Thus, in some embodiments, the modified immune cells provided herein may comprise the Cbl-b inhibitors described herein, for example, comprising the Cbl-b inhibitors described herein in the cytoplasm of the modified immune cells.

[0403] In some cases, the present invention provides isolated modified immune cells, wherein the modified immune cells have been contacted with a Cbl-b inhibitor or a composition thereof described herein or with a Cbl-b inhibitor or a composition thereof described herein. In some embodiments, the modified immune cells are T cells, B cells, or natural killer (NK) cells. In some embodiments, the modified immune cells are hematopoietic cells, pluripotent stem cells, myeloid progenitor cells, lymphoid progenitor cells, T cells, B cells, or NK cells.

[0404] In some embodiments of the modified immune cells of the separation, the modified immune cells are T cells, and the T cells show increased T cell activation, increased T cell proliferation, reduced T cell exhaustion and / or reduced T cell tolerance. In some embodiments, the increased T cell activation comprises increasing the generation of one or more cytokines of T cells or peripheral immune cells (e.g., myeloid cells) from the activated T cell microenvironment. In some embodiments, the one or more cytokines include but are not limited to: IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-13, IL-18, TNFα and GM-CSF. In some embodiments, the cytokine is one or more of the following: IL-2, IFN-γ, TNFα and GM-CSF. In some embodiments, the cytokine is a chemokine. In some embodiments, the one or more chemokines include but are not limited to: IP-10, eotaxin, GROα, RANTES, MIP-1α, MIP-1β, MIP-2, MCP-1 and MCP-3. In some embodiments, increased T cell activation comprises increasing the cell surface expression of one or more T cell activation markers. In some embodiments, the one or more T cell activation markers include but are not limited to: CD25, CD44, CD62L, CD69, CD152 (CTLA4), CD154, CD137 and CD279. In some embodiments, the one or more T cell activation markers include but are not limited to: CD25, CD69 and CTLA4. In some embodiments, the T cell activation marker is CD25 and / or CD69. In some embodiments, the T cells have been contacted with anti-CD3 antibodies or with anti-CD3 antibodies. In some embodiments, the T cells have been contacted with a combination of anti-CD3 antibodies and anti-CD28 antibodies or with a combination of anti-CD3 antibodies and anti-CD28 antibodies.

[0405] In some embodiments of the modified immune cells of the separation, the modified immune cells are NK cells, and the NK cells show increased NK cell activation. In some embodiments, the increased NK cell activation comprises one or more cytokines (e.g., IFN-γ, TNFα and / or MIP-1β) secretion increase. In some embodiments, the increased NK cell activation comprises one or more NK cell activation markers (e.g., CD69 and / or CD107a) cell surface expression increase.

[0406] In some embodiments of the modified immune cell of the separation, the modified immune cell is a B cell, and the B cell shows increased B cell activation. In some embodiments, the increased B cell activation comprises an increase in the cell surface expression of one or more B cell activation markers (e.g., CD69, CD86 and / or HLA-DR).

[0407] In some of any embodiments of the methods or modified immune cells provided herein, the immune cell or modified immune cell is a mammalian cell (e.g., a human cell). In some embodiments, the immune cell or modified immune cell is a human cell.

[0408] In some cases, incubation is performed according to techniques such as those described in US Pat. No. 6,040,177; Klebanoff et al., J Immunother., 35:651-660, 2012; Terakura et al., Blood, 119:72-82, 2012; and Wang et al., J Immunother., 35:689-701, 2012.

[0409] The immune cells to be modified or modified immune cells provided by the present invention can be engineered to express a recombinant chimeric receptor, such as a chimeric antigen receptor (CAR). In some embodiments, CAR comprises from its N-terminus to its C-terminus: an extracellular ligand binding domain, a transmembrane domain, an intracellular co-stimulatory domain, and an activation cytoplasmic signaling domain. In some embodiments, CAR comprises from its N-terminus to its C-terminus: an extracellular ligand binding domain, a transmembrane domain, and an activation cytoplasmic signaling domain. The immune cells can be engineered to express a recombinant chimeric receptor (e.g., CAR) before, during, or after contact with the Cbl-b inhibitor provided by the present invention. In some embodiments, the immune cells to be modified are T cells (e.g., CD4 + T cells or CD8 +T cells). In further embodiments, the T cells comprise a recombinant chimeric receptor, such as a CAR. In some embodiments, the modified immune cells are modified T cells (e.g., CD4 + T cells or CD8 + T cells). In a further embodiment, the modified T cells comprise a recombinant chimeric receptor, such as a CAR. Methods for generating immune cells expressing recombinant chimeric receptors are well known in the art, for example, by introducing a nucleic acid encoding the recombinant chimeric receptor (e.g., CAR) into immune cells (e.g., T cells) via a vector (e.g., a viral vector). See, for example, International Patent Application No. WO2017 / 096329 and U.S. Publication No. US 2017 / 0204372.

[0410] In particular, the present invention provides a method for generating an expanded lymphocyte population, the method comprising: (a) obtaining a biological sample comprising lymphocytes from an individual suffering from cancer, wherein the individual has received or is receiving an effective amount of a Cbl-b inhibitor as a monotherapy or as part of a combination therapy, and (b) culturing lymphocytes in a cell culture medium comprising at least one T cell growth factor to generate an expanded lymphocyte population. In some embodiments, the lymphocytes are tumor infiltrating lymphocytes (TIL). In some embodiments, the lymphocytes are TILs that have been isolated from or are isolated from a tumor of a mammalian subject suffering from cancer. In other embodiments, the lymphocytes are peripheral blood mononuclear cells (PBMCs). In some embodiments, the at least one T cell growth factor comprises one or more of the group consisting of IL-2, IL-7, IL-15, and IL-21, optionally wherein the at least one T cell growth factor comprises IL-2. In some embodiments, the cell culture medium further comprises an anti-CD3 antibody, or both an anti-CD3 antibody and an anti-CD28 antibody. In some embodiments, the cell culture medium further comprises a Cbl-b inhibitor. In some embodiments, the cell culture medium further comprises irradiated feeder cells. In some embodiments, the individual is a human patient. The present invention also provides a composition comprising an expanded TIL population generated by the above method and a physiologically acceptable buffer.

[0411] In some embodiments, the method of isolating and processing immune cells to be modified or modified (i.e., modified immune cells) includes a step of freezing (e.g., cryopreserving) the cells before or after isolation, incubation (e.g., incubation with a Cbl-b inhibitor) and / or engineering (e.g., introducing a nucleic acid encoding a recombinant chimeric receptor into the immune cells). A variety of freezing solutions and parameters known in the art can be used.

[0412] B. Adoptive Cell Therapy

[0413] The modified immune cells, such as the expanded lymphocyte populations or compositions thereof generated by the methods described herein, can be used as therapeutic agents in methods for treating individuals in need thereof (e.g., individuals suffering from cancer). Such treatment methods include adoptive cell therapy. In some embodiments, the treatment method includes isolating cells from an individual, preparing, treating, culturing, and / or engineering them, as described herein, and reintroducing them into the same individual before or after cryopreservation. In some embodiments, the treatment method includes isolating cells from an individual, preparing, treating, culturing, and / or engineering them, as described herein, and reintroducing them into a different individual before or after cryopreservation.

[0414] Therefore, in some cases, the present invention provides a method for regulating an immune response in an individual, the method comprising administering an effective amount of a modified immune cell of the present invention or a composition thereof to an individual in need (e.g., an individual with T cell dysfunction). In some embodiments, the individual suffers from cancer. In some embodiments, the present invention provides a method for treating a cancer that responds to Cbl-b activity inhibition, the method comprising administering an effective amount of a modified immune cell of the present invention or a composition thereof to an individual suffering from the cancer that responds to Cbl-b activity inhibition. In some embodiments, the present invention provides a method for inhibiting abnormal cell proliferation, the method comprising administering an effective amount of a modified immune cell of the present invention or a composition thereof to an individual in need. As used herein, the term "abnormal cell proliferation" includes hyperplasia or cancer cell proliferation. Cancer cells may be derived from hematological cancer or non-hematological cancer. In some embodiments, the cancer is a hematological cancer, such as a lymphoma, leukemia, or myeloma. In other embodiments, the cancer cells are derived from non-hematological cancers, such as sarcoma, carcinoma, or melanoma.

[0415] In certain embodiments, a composition comprising a range of about 1 million to about 100 billion cells, e.g., about 1 million to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or a range defined by any two of the foregoing values), e.g., about 10 million to about 100 billion cells (e.g., about 20 million cells, about 30 million cells, about 40 million cells, about 60 million cells, about 70 million cells, about 80 million cells, about 90 million cells, about 100 million cells), or about 100 billion cells. In some cases, the modified immune cells provided herein have a population size of about 70 million cells, about 80 million cells, about 90 million cells, about 10 billion cells, about 25 billion cells, about 50 billion cells, about 75 billion cells, about 90 billion cells, or a range defined by any two of the foregoing values), and in some cases, about 100 million cells to about 50 billion cells (e.g., about 120 million cells, about 250 million cells, about 350 million cells, about 450 million cells, about 650 million cells, about 800 million cells, about 900 million cells, about 3 billion cells, about 30 billion cells, about 45 billion cells), or any value in between these ranges.

[0416] The modified immune cells and compositions thereof can be administered using standard administration techniques, formulations and / or devices. The present invention provides formulations and devices for storage and administration of the compositions, such as syringes and vials. Preparations or pharmaceutical compositions comprising the modified immune cells include those for intravenous, intraperitoneal, subcutaneous or intramuscular administration. In some embodiments, the modified immune cells are administered parenterally. As used herein, the term "parenteral" includes but is not limited to intravenous, intramuscular, subcutaneous and intraperitoneal administration. In some embodiments, the cell population is administered to a subject using peripheral systemic delivery by intravenous, intraperitoneal or subcutaneous injection. The compositions of the modified immune cells can be provided as sterile liquid preparations, such as isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions, which in some cases can be buffered to a selected pH value. Viscous compositions can be formulated within a suitable viscosity range to provide a longer contact time with a specific tissue. Liquid or viscous compositions may include a carrier, which may be a solvent or dispersion medium comprising, for example, water, saline, phosphate buffered saline, a polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. Sterile injectable solutions may be prepared by incorporating the modified immune cells into a solvent, for example, by mixing with a suitable carrier, diluent, or excipient (e.g., sterile water, physiological saline, glucose, dextrose, or the like).

[0417] In some embodiments, the modified immune cells are co-administered with one or more additional therapeutic agents or another therapeutic intervention, either simultaneously or sequentially in any order. For example, in some treatment regimens of the present invention, both the modified immune cells and a Cbl-b inhibitor are administered to a mammalian subject in need thereof, wherein the Cbl-b inhibitor is a compound of Formula (I), (IA)-(IN), (II-A)-(II-L), (III-A)-(III-D), (IV-A)-(IV-C), (VA)-(VG), or (VI-A)-(VI-D), or any variant thereof. Thus, in some embodiments, the treatment regimen comprises both adoptive cell therapy and chemotherapy.

[0418] After the modified immune cell is applied to an individual (e.g., people), the bioactivity of the modified immune cell group can be determined by methods known in the art. The parameter to be evaluated includes the specific binding of modified immune cells or other immune cells in vivo (e.g., by imaging) or in vitro (e.g., by ELISA or flow cytometry) to antigens. In some embodiments, cytotoxicity assays can be used to determine the ability of modified immune cells to destroy target cells (see, e.g., Kochenderfer et al., J. Immunotherapy, 32: 689-702, 2009; and Herman et al., J. Immunological Methods, 285: 25-40, 2004). In some embodiments, the bioactivity of the modified immune cell can also be determined by measuring the expression and / or secretion of certain cytokines (e.g., IL-2 and IFNγ).

[0419] C. Administration of Cbl-b Inhibitors

[0420] In some cases, a Cbl-b inhibitor or composition thereof can be directly administered to an individual to modulate an immune response, treat a disease or condition (e.g., cancer and / or abnormal cell proliferation), and / or inhibit Cbl-b activity in the individual. The Cbl-b inhibitor can be a compound shown in Table 1 and / or Table 1A, a tautomer, a stereoisomer, or a pharmaceutically acceptable salt thereof.

[0421] In some embodiments, the present invention provides a method for regulating an immune response, comprising administering to an individual an effective amount of a Cbl-b inhibitor provided herein or a composition thereof to regulate the immune response of the individual. In some embodiments, the individual suffers from cancer.

[0422] In some embodiments, the present invention provides a method for treating cancer responsive to inhibition of Cbl-b activity, comprising administering to a subject an effective amount of a Cbl-b inhibitor provided herein or a composition thereof to treat the cancer responsive to inhibition of Cbl-b activity.

[0423] In some embodiments, the present invention provides a method for inhibiting abnormal cell proliferation (eg, hyperplasia), comprising administering to a subject an effective amount of a Cbl-b inhibitor provided herein or a composition thereof to inhibit abnormal cell proliferation in the subject.

[0424] In some embodiments, the present invention provides a method for inhibiting Cbl-b activity, comprising administering to a subject an effective amount of a Cbl-b inhibitor provided herein or a composition thereof to inhibit Cbl-b activity in the subject.

[0425] In some embodiments, for example, in modulating an immune response in a subject in need thereof (e.g., a subject with T cell dysfunction), treating a disease or condition in a subject (e.g., a cancer and / or abnormal cell proliferation in a subject), and / or inhibiting Cbl-b activity in a subject, the appropriate dose of the active agent will depend on the type of condition, disease, or condition to be treated (as defined above), the severity and course of the condition, disease, or condition, whether the agent is administered for preventive or therapeutic purposes, previous treatments, the subject's clinical history and response to the Cbl-b inhibitor, and the discretion of the attending physician. The Cbl-b inhibitor or composition thereof is appropriately administered to the subject once or over a series of treatments. In some embodiments, the treatment comprises multiple administrations of the Cbl-b inhibitor or composition thereof, wherein the intervals between administrations can vary. For example, the interval between the first and second administrations is about 1 month, and the interval between subsequent administrations is about 3 months. In some embodiments, the Cbl-b inhibitor is administered at a fixed dose. In some embodiments, the Cbl-b inhibitor is administered to the subject at a fixed dose (e.g., mg / kg) based on the subject's body weight.

[0426] In some embodiments of the present invention, the cancer is a hematological cancer. For example, the hematological cancer can be lymphoma, leukemia, or myeloma. In other embodiments of the present invention, the cancer is a non-hematological cancer. In particular, the non-hematological cancer can be carcinoma, sarcoma, or melanoma.

[0427] In some embodiments, the Cbl-b inhibitor is co-administered with one or more additional therapeutic agents or another therapeutic intervention, either simultaneously or sequentially in any order. For example, in some therapeutic regimens of the present invention, both the Cbl-b inhibitor and the modified immune cells are administered to a mammalian subject in need thereof, wherein the Cbl-b inhibitor is a compound of Formula (I), (IA)-(IN), (II-A)-(II-L), (III-A)-(III-D), (IV-A)-(IV-C), (VA)-(VG), or (VI-A)-(VI-D), or any variant thereof. The Cbl-b inhibitor can be a compound of Table 1 and / or Table 1A, a tautomer thereof, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. Thus, in some embodiments, the therapeutic regimen comprises both adoptive cell therapy and chemotherapy.

[0428] In some embodiments, the effectiveness of administering a Cbl-b inhibitor in a method of the present invention (e.g., a method of modulating an immune response in a subject) can be assessed by measuring the biological activity of immune cells present in a sample (e.g., a blood sample) isolated from the treated subject. For example, the efficacy of treatment can be assessed by measuring the ability of immune cells isolated from a subject to destroy target cells in a cytotoxicity assay following treatment with a Cbl-b inhibitor. In some embodiments, the biological activity of immune cells present in a sample (e.g., a blood sample) can be assessed by measuring the expression and / or secretion of certain cytokines (e.g., IL-2 and IFNγ).

[0429] The present invention provides a method for treating cancer, comprising administering an effective amount of a Cbl-b inhibitor to a subject having cancer, and administering an effective amount of an additional therapeutic agent to the subject. The present invention also provides a method for treating a subject having cancer, comprising administering an effective amount of a Cbl-b inhibitor to the subject, and administering an effective amount of an additional therapeutic agent to the subject. Furthermore, the present invention provides a method for increasing an anti-cancer immune response, comprising administering an effective amount of a Cbl-b inhibitor to a subject having cancer, and administering an effective amount of an additional therapeutic agent to the subject. The present invention also provides a method for treating cancer, comprising administering an effective amount of a Cbl-b inhibitor to a subject having cancer, wherein the subject has received or is currently receiving an effective amount of an additional therapeutic agent.

[0430] In some embodiments of the methods of the preceding paragraphs, the Cbl-b inhibitor and the additional therapeutic agent are administered sequentially in either order. As used herein, the terms "sequentially," "continuously," and "sequentially" refer to administering the Cbl-b inhibitor after administering the additional therapeutic agent, or administering the additional therapeutic agent after administering the Cbl-b inhibitor. For example, sequential administration may comprise administering the Cbl-b inhibitor in the absence of an additional therapeutic agent during an induction phase (primary treatment), followed by a post-induction treatment phase comprising administering the additional therapeutic agent. The method may further comprise a maintenance phase comprising administering the Cbl-b inhibitor or the additional therapeutic agent. Alternatively, sequential administration may comprise administering the additional therapeutic agent in the absence of a Cbl-b inhibitor during an induction phase (primary treatment), followed by a post-induction treatment phase comprising administering the Cbl-b inhibitor. The method may further comprise a maintenance phase comprising administering the Cbl-b inhibitor or the additional therapeutic agent.

[0431] In some embodiments of the combination therapy method, the Cbl-b inhibitor and the additional therapeutic agent are administered simultaneously. As used herein, the terms "simultaneously," "concurrently," and "in parallel" refer to administering the Cbl-b inhibitor and the additional therapeutic agent during the same doctor's visit or during the same treatment phase. For example, the Cbl-b inhibitor and the additional therapeutic agent can be administered during one or more of the induction phase, the treatment phase, and the maintenance phase. However, simultaneous administration does not require that the Cbl-b inhibitor and the additional therapeutic agent be present together in a single formulation or pharmaceutical composition, or that the Cbl-b inhibitor and the additional therapeutic agent be administered at precisely the same time.

[0432] 1. Combination therapy including Cbl-b inhibitors and immune checkpoint inhibitors

[0433] In some embodiments of the combined treatment methods of the present invention for treating cancer, the additional therapeutic agent comprises an immune checkpoint inhibitor. The term "immune checkpoint" refers to a signaling pathway that prevents immune cell activation, while the term "immune checkpoint inhibitor" refers to a compound that blocks immune checkpoints to release the brakes on immune cell activation. In some embodiments, the immune checkpoint inhibitor is an antagonist of at least one inhibitory checkpoint molecule. In some embodiments, the inhibitory checkpoint molecule is selected from the group consisting of: PD-1 (CD279), PD-L1 (CD274), CTLA-4 (CD125), LAG3 (CD223), PVR (CD155), PVRL2 (CD112), PVRL3 (CD113), TIGIT, TIM3 (CD366), and VISTA. In some embodiments, the immune checkpoint inhibitor is an antagonist of at least one inhibitory checkpoint molecule, and the at least one inhibitory checkpoint molecule is selected from the group consisting of PD-1 (CD279), PD-L1 (CD274), and CTLA-4 (CD152).

[0434] PD-1 refers to programmed cell death protein 1 (PD-1). PD-1 antagonists suitable for use in the methods, medicaments, and uses of the present invention include any chemical compound or biological molecule that blocks the binding of PD-L1 expressed on cancer cells or antigen-presenting cells to PD-1 expressed on lymphocytes (T cells, B cells, and / or NK cells). Alternative names or synonyms for PD-1 and its ligands include: CD279, PDCD1, PD1, and SLEB2 for PD-1; and CD274, PDCD1L1, PDL1, B7H1, B7-4, and B7-H for programmed cell death 1 ligand 1 (PD-L1). In some embodiments of treating human subjects, the PD-1 antagonist blocks the binding of human PD-L1 to human PD-1. The amino acid sequence of the mature form of human PD-1 is shown as residues 21-288 in NCBI locus number NP_005009. The amino acid sequence of the mature form of human PD-L1 is shown as residues 19-290 in NCBI locus accession number NP_054862.

[0435] CTLA-4 refers to cytotoxic T lymphocyte-associated protein 4. CTLA-4 antagonists suitable for use in the methods, medicaments, and uses of the present invention include blocking the binding of CTLA-4 expressed on lymphocytes (T cells, B cells, and / or NK cells) to ligands (CD80 and / or CD86) expressed on antigen-presenting cells. Alternative names or synonyms for CTLA-4 include: CD152, CTLA4, ALPS5, CELIAC3, GRD4, GSE, and IDDM12. In some embodiments for treating human subjects, CTLA-4 antagonists block the binding of human CTLA-4 to human ligands. The amino acid sequence of the mature form of human CTLA-4 is shown as residues 36-223 in NCBI locus number NP_005205.

[0436] LAG3 refers to lymphocyte activation gene 3 protein. LAG3 antagonists suitable for use in the methods, medicaments, and uses of the present invention include any chemical compound or biomolecule that blocks the binding of LAG3 expressed on lymphocytes (T cells, B cells, and / or NK cells) to ligands (MHC class II) expressed on antigen-presenting cells. LAG3 is also known as CD223. In some embodiments for treating human subjects, LAG3 antagonists block the binding of human LAG3 to human ligands. The amino acid sequence of the mature form of human LAG3 is shown in NCBI Locus No. NP_002277, residues 23-525.

[0437] PVR refers to the poliovirus receptor. PVR antagonists suitable for the methods of treatment, medicines and uses of the present invention include any chemical compound or biomolecule that blocks the binding of PVR expressed on cancer cells or antigen-presenting cells to TIGIT expressed on lymphocytes (T cells, B cells, and / or NK cells). Alternative names or synonyms for PVR include CD155, PVS, HVED, NECL5, nectin-like protein 5, and TAGE4. In some embodiments of treating human subjects, PVR antagonists block the binding of human PVR to human TIGIT. Human PVR has multiple isoforms. The amino acid sequence of the α isoform of human PVR is shown in NCBI locus number NP_006496. The amino acid sequence of the β isoform of human PVR is shown in NCBI locus number NP_001129240. The amino acid sequence of the γ isoform of human PVR is shown in NCBI locus number NP_001129241. The amino acid sequence of the delta isoform of human PVR is shown in NCBI locus accession number NP_001129242.

[0438] PVRL2 refers to poliovirus receptor-related 2. PVRL2 antagonists suitable for use in the methods of treatment, medicaments, and uses of the present invention include any chemical compound or biological molecule that blocks the binding of PVRL2 expressed on cancer cells or antigen-presenting cells to TIGIT expressed on lymphocytes (T cells, B cells, and / or NK cells). Alternative names or synonyms for PVRL2 include: CD112, NECTIN2, HVEB, herpesvirus entry mediator B, PRR2, and PVRR2. In some embodiments for treating human subjects, the PVRL2 antagonist blocks the binding of human PVRL2 to human TIGIT. The amino acid sequence of the alpha isoform of human PVRL2 is shown in NCBI locus number NP_002847. The amino acid sequence of the delta isoform of human PVRL2 is shown in NCBI locus number NP_001036189.

[0439] PVRL3 refers to poliovirus receptor-related 3. PVRL3 antagonists suitable for use in the methods of treatment, medicaments, and uses of the present invention include any chemical compound or biological molecule that blocks the binding of PVRL3 expressed on cancer cells or antigen-presenting cells to TIGIT expressed on lymphocytes (T cells, B cells, and / or NK cells). Alternative names or synonyms for PVRL3 include: CD113, NECTIN3, PRR3, and PVRR3. In some embodiments for treating human subjects, the PVRL3 antagonist blocks the binding of human PVRL3 to human TIGIT. The amino acid sequence of human PVRL3 isoform 1 is shown in NCBI locus number NP_056295. The amino acid sequence of isoform 2 of human PVRL3 is shown in NCBI locus number NP_001230215. The amino acid sequence of isoform 3 of human PVRL3 is shown in NCBI locus number NP_001230217.

[0440] TIGIT refers to a T cell immunoreceptor with Ig and ITIM domain proteins. TIGIT antagonists suitable for the treatment methods, drugs and uses of the present invention include any chemical compound or biological molecule that blocks the binding of TIGIT expressed on lymphocytes (T cells, B cells or NK cells) to ligands (CD112, CD113 and / or CD155) expressed on cancer cells or antigen presenting cells. Alternative names or synonyms for TIGIT include: VSIG9, V-set and immunoglobulin domains containing 9, VSTM3, V-set and transmembrane domains containing 3, and Washington University cell adhesion molecule (WUCAM). In some embodiments for treating human subjects, TIGIT antagonists block the binding of human TIGIT to human ligands. The amino acid sequence of the mature form of human TIGIT is shown in residues 22-244 in NCBI locus number: NP_776160.

[0441] TIM3 refers to T-cell immunoglobulin and mucin-domain containing-3 protein. TIM3 antagonists suitable for use in the treatment methods, medicaments, and uses of the present invention include any chemical compound or biomolecule that blocks the binding of TIM3 expressed on lymphocytes (T cells, B cells, or NK cells) to its ligand (galectin-9 phosphatidylserine) expressed on antigen-presenting cells. Alternative names or synonyms for TIM3 include CD366, HAVCR2, hepatitis A virus cellular receptor 2, KIM3, and SPTCL. In some embodiments for treating human subjects, the TIM3 antagonist blocks the binding of human TIM3 to its human ligand. The amino acid sequence of the mature form of human TIM3 is shown in NCBI locus number NP_116171, residues 22-301.

[0442] VISTA refers to the V domain Ig inhibitory factor of T cell activation. VISTA antagonists suitable for the methods of treatment, medicines and uses of the present invention include any chemical compound or biomolecule that blocks the binding of VISTA expressed on lymphocytes (T cells, B cells and / or NK cells) to ligands expressed on cancer cells or antigen-presenting cells. Alternative names or synonyms for VISTA include: VSIR, V-set immunomodulatory receptor, PD-1H, B7H5, GI24, PP2135, SISP1 and Dies1. In some embodiments of treating human subjects, VISTA antagonists block the binding of human VISTA to human ligands. The amino acid sequence of mature human VISTA is shown in residues 33-311 of NCBI locus number: NP_071436.

[0443] Immune checkpoint inhibitors can be biomolecules. For example, immune checkpoint inhibitors may comprise antibodies or their antigen-binding fragments. The antibody or fragment may be a monoclonal antibody (mAb), such as a human antibody, a humanized antibody, or a chimeric antibody, and may include a human constant region. In some embodiments, the human constant region is selected from the group consisting of IgG1, IgG2, IgG3, and IgG4 constant regions, and in certain embodiments, the human constant region is an IgG1 or IgG4 constant region. In some embodiments, the antibody or fragment is a bispecific antibody. In some embodiments, the antigen-binding fragment comprises one of the group consisting of Fab, Fab'-SH, F(ab')2, scFv, and Fv fragments.

[0444] In some embodiments, the at least one inhibitory checkpoint molecule comprises PD-1. In some embodiments, the immune checkpoint inhibitor is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, and biosimilars thereof. In one embodiment, the anti-PD-1 antibody is pembrolizumab (MK-3475, manufactured by Merck & Co. In one embodiment, the anti-PD-1 antibody is nivolumab (BMS-936558 or MDX-1106, marketed by Bristol-Myers Squibb as In one embodiment, the anti-PD-1 antibody is cemiplimab (REGN2810, Regeneron). In some embodiments, the immune checkpoint inhibitor is pembrolizumab, nivolumab, or a variant of cemiplimab.

[0445] In some embodiments, the at least one inhibitory checkpoint molecule comprises PD-L1. In some embodiments, the immune checkpoint inhibitor is selected from the group consisting of atezolizumab, avelumab, durvalumab, and biosimilars thereof. In one embodiment, the anti-PD-L1 antibody is atezolizumab (available from Genentech, Inc. as In one embodiment, the anti-PD-L1 antibody is avelumab (marketed by EMD Serono, Inc. and Pfizer, Inc.). In one embodiment, the anti-PD-L1 antibody is durvalumab (MEDI4736, marketed by AstraZeneca as In some embodiments, the immune checkpoint inhibitor is a variant of atezolizumab, avelumab, or durvalumab.

[0446] In some embodiments, the at least one inhibitory checkpoint molecule comprises CTLA-4. In some embodiments, the immune checkpoint inhibitor is selected from the group consisting of ipilimumab, tremelimumab, and biosimilars thereof. In one embodiment, the anti-CTLA4 antibody is ipilimumab (MDX-010 or BMS-734016, marketed by Bristol-Myers Squibb as In one embodiment, the anti-CTLA4 antibody is tremelimumab (ticilimumab, CP-675,206, developed by AstraZeneca). In some embodiments, the immune checkpoint inhibitor is ipilimumab or a variant of tremelimumab.

[0447] In some embodiments, the monoclonal antibody is a "variant" antibody comprising heavy and light chain sequences identical to those in a "reference" antibody, except that 3, 2 or 1 conservative amino acid substitutions are present at positions outside the light chain CDR and / or 6, 5, 4, 3, 2 or 1 conservative amino acid substitutions are present at positions outside the heavy chain CDR (e.g., variant positions are all located in the framework region or constant region). In other words, the reference antibody and the variant antibody both comprise the same CDR sequence, but differ from each other by having conservative amino acid substitutions at no more than 3 or 6 other positions in their full-length light and heavy chain sequences, respectively. The variant antibody is substantially the same as the reference antibody in terms of binding affinity for the inhibitory checkpoint molecule and the ability to block the binding of the inhibitory checkpoint molecule to its ligand.

[0448] In other embodiments, the immune checkpoint inhibitor may comprise an immunoadhesin comprising an inhibitory checkpoint molecule binding domain of one of its ligands fused to a constant region (e.g., an Fc region of an immunoglobulin molecule).

[0449] As used herein, the term "biosimilar" or "biosimilar" refers to a biological product that is similar to a reference product approved by the U.S. Federal Drug Administration (FDA) but has no clinically significant differences in safety and efficacy. For example, there may be differences between a biosimilar product and a reference product in clinically inactive ingredients (e.g., differences in formulation excipients, subtle differences in glycosylation, etc.). Clinically significant characteristics can be assessed through pharmacokinetic and pharmacodynamic studies. In some embodiments, the biosimilar product is an interchangeable product determined by the FDA.

[0450] 2. Combination therapy comprising Cbl-b inhibitors and anti-tumor agents

[0451] In some embodiments of the combined treatment methods for treating cancer of the present invention, the additional therapeutic agent comprises an anti-tumor agent. As used herein, the terms "antineoplastic agent" and "antineoplastic drug" refer to therapeutic agents classified according to the Anatomical Therapeutic Chemical Classification system (ATC) code L01 developed by the World Health Organization. In some embodiments, the anti-tumor agent is classified as one of the group consisting of cytotoxic antibiotics (ATC code L01D), plant alkaloids (ATC code L01C), antimetabolites (ATC code L01B), alkylating agents (ATC code L01A), and other anti-tumor agents (ATC code L01X). In some embodiments, the anti-tumor agent is a small molecule drug (e.g., a cancer chemotherapeutic agent) rather than a biological molecule.

[0452] Cytotoxic antibiotics are antitumor agents suitable for use in the methods, medicines, and uses of the present invention. In some embodiments, the cytotoxic antibiotic is selected from the group consisting of ixabepilone, mitomycin, plicamycin, bleomycin, pixantrone, amrubicin, valrubicin, pirarubicin, mitoxantrone, idarubicin, zorubicin, aclarubicin, epirubicin, daunorubicin, doxorubicin, and dactinomycin.

[0453] Plant alkaloids are antitumor agents suitable for use in the methods, medicines, and uses of the present invention. In some embodiments, the plant alkaloids are selected from the group consisting of trabectedin, cabazitaxel, paclitaxel poliglumex, docetaxel, paclitaxel, demecolcine, teniposide, etoposide, vintafolide, vinflunine, vinorelbine, vindesine, vincristine, and vinblastine.

[0454] Antimetabolites are antitumor agents suitable for use in the methods, medicaments, and uses of the present invention. In some embodiments, the antimetabolite is a pyrimidine analog, a purine analog, or a folic acid analog. In some embodiments, the antimetabolite is selected from the group consisting of floxuridine, trifluridine, tegafur, fluorouracil, decitabine, azacitidine, capecitabine, gemcitabine, carmofur, tegafur, fluorouracil, cytarabine, nelarabine, clofarabine, fludarabine, cladribine, tioguanine, mercaptopurine, pralatrexate, pemetrexed, raltitrexed, and methotrexate.

[0455] Alkylating agents are suitable antitumor agents that can be used in the methods of treatment, medicines and uses of the present invention. In some embodiments, the alkylating agent is selected from the group consisting of dacarbazine, temozolomide, pipobroman, mitobronitol, etoglucid, uracil mustard, mustard, ranimustine, nimustine, fotemustine, streptozocin, semustine, lomustine, carmustine, carboquone, triaziquone, thiotepa, mannosulfan, treosulfan, busulfan, bendamustine, prednimustine, trofosfamide, ifosfamide, mechlorethamine, melphalan, chlorambucil, and cyclophosphamide.

[0456] In other embodiments, the anti-tumor agent comprises an agent selected from the group consisting of platinum compounds (ATC code L01XA), methylhydrazine (ATC code L01XB), sensitizers (ATC code L01XD), protein kinase inhibitors (ATC code L01XE), and other anti-tumor agents (ATC code L01XA).

[0457] Platinum compounds are suitable antitumor agents for use in the methods of treatment, medicaments, and uses of the present invention. In some embodiments, the platinum compound is selected from the group consisting of cisplatin, carboplatin, oxaliplatin, satraplatin, and polyplatillen.

[0458] 3. Combination therapy including Cbl-b inhibitors and radiotherapy

[0459] The present invention provides a method for treating cancer, comprising administering an effective amount of a Cbl-b inhibitor to a subject suffering from cancer, and administering an effective amount of radiation therapy to the subject. The present invention also provides a method for treating a subject suffering from cancer, comprising administering an effective amount of Cbl-b to the subject, and administering an effective amount of radiation therapy to the subject. Furthermore, the present invention provides a method for increasing an anti-cancer immune response, comprising administering an effective amount of a Cbl-b inhibitor to a subject suffering from cancer, and administering an effective amount of radiation therapy to the subject. The present invention also provides a method for treating cancer, comprising administering an effective amount of a Cbl-b inhibitor to a subject suffering from cancer, wherein the subject has received or is currently receiving an effective amount of radiation therapy.

[0460] In some embodiments, the radiation therapy is external beam radiation therapy. In other embodiments, the radiation therapy is internal beam radiation therapy. In some embodiments, the radiation therapy is ablative radiation therapy.

[0461] In some embodiments, the combination therapy regimens of the present invention comprise administration of a Cbl-b inhibitor, radiation therapy, and one or both of an immune checkpoint inhibitor and an anti-tumor agent.

[0462] The present invention provides a method for treating cancer, comprising administering to an individual suffering from cancer a combination therapy comprising an effective amount of a Cbl-b inhibitor and an effective amount of a cancer vaccine. The present invention also provides a medicament comprising a Cbl-b inhibitor for use in combination with a cancer vaccine to treat cancer, as well as a medicament for treating cancer comprising both a Cbl-b inhibitor and a cancer vaccine. The present invention further provides the use of a Cbl-b inhibitor in the preparation of a medicament for treating cancer in an individual when administered in combination with a cancer vaccine. The present invention further provides the use of a Cbl-b inhibitor and a cancer vaccine in the preparation of a medicament for treating cancer. In some embodiments, the Cbl-b inhibitor is a "small molecule."

[0463] The present invention also provides a method for treating cancer, comprising administering to an individual with cancer a combination therapy comprising an effective amount of a Cbl-b inhibitor and an effective amount of an oncolytic virus. The present invention also provides a drug comprising a Cbl-b inhibitor for use in combination with an oncolytic virus to treat cancer, and a drug for treating cancer comprising both a Cbl-b inhibitor and an oncolytic virus. The present invention further provides the use of a Cbl-b inhibitor in the preparation of a drug for treating individual cancer when administered in combination with an oncolytic virus. The present invention further provides the use of a Cbl-b inhibitor and an oncolytic virus in the preparation of a drug for treating cancer. In some embodiments, the Cbl-b inhibitor is a "small molecule".

[0464] In some embodiments of the methods of treatment, medicaments and uses of the present invention, the cancer is a hematological cancer, such as lymphoma, leukemia or myeloma. In other embodiments of the methods of treatment, medicaments and uses of the present invention, the cancer is a non-hematological cancer, such as sarcoma, carcinoma or melanoma.

[0465] Hematological cancers include, but are not limited to, one or more leukemias, such as B-cell acute lymphoblastic leukemia ("BALL"), T-cell acute lymphoblastic leukemia ("TALL"), acute lymphoblastic leukemia (ALL); one or more chronic leukemias, including, but not limited to, chronic myeloid leukemia (CML) and chronic lymphocytic leukemia (CLL); other hematological cancers or hematological diseases, including, but not limited to, B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma (BCL), and leukemia. The term "preleukemia" refers to a variety of hematologic disorders characterized by ineffective production (or dysplasia) of myeloid blood cells, including diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, follicular small cell lymphoma or follicular large cell lymphoma, malignant lymphoproliferative disorders, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndrome, non-Hodgkin lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom macroglobulinemia, and "preleukemias," which are a group of hematologic disorders characterized by ineffective production (or dysplasia) of myeloid blood cells.

[0466] Non-hematological cancers include, but are not limited to, neuroblastoma, renal cell carcinoma, colon cancer, colorectal cancer, breast cancer, epithelial squamous cell carcinoma, melanoma, gastric cancer, brain cancer, lung cancer (e.g., NSCLC), pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, prostate cancer, testicular cancer, thyroid cancer, uterine cancer, adrenal cancer, and head and neck cancer.

[0467] Activation threshold lowering agents or costimulation demand lowering agents, such as Cbl-b inhibitors, are used for the treatment of cancer. The effectiveness of the treatment of cancer is determined by assessing clinical outcomes (e.g., tumor size or tumor number reduction and / or survival rate). In some embodiments, "treating cancer" includes assessing the patient's response to the treatment regimen according to the response evaluation criteria (RECIST 1.1 version) in the solid tumor (see, e.g., Eisenhauer et al., Eur J Cancer, 45: 228-247, 2009; and Nishino et al., Am J Roentgenol, 195: 281-289, 2010). The response criteria for determining the anti-tumor response of interest according to RECIST 1.1 include complete remission (CR); partial remission (PR); progression of disease (PD); and stable disease (SD).

[0468] The present invention also provides a method for treating cancer, which comprises administering a combination therapy to an individual suffering from cancer, wherein the combination therapy comprises an effective amount of an agent (activation threshold lowering agent) that reduces the activation threshold of immune cells (e.g., T cells, B cells and / or NK cells) and an effective amount of a cancer vaccine. The present invention also provides a drug comprising an activation threshold lowering agent used in combination with a cancer vaccine to treat cancer, and a drug for treating cancer comprising both an activation threshold lowering agent and a cancer vaccine. The present invention further provides the use of an activation threshold lowering agent in the preparation of a drug for treating individual cancer when administered in combination with a cancer vaccine. The present invention further provides the use of an activation threshold lowering agent and a cancer vaccine in the preparation of a drug for treating cancer.

[0469] The present invention also provides a method for treating cancer, which comprises administering a conjoint therapy to an individual with cancer, wherein the conjoint therapy comprises an effective amount of an agent (activation threshold lowering agent) for reducing the activation threshold of immune cells (e.g., T cells, B cells and / or NK cells) and an effective amount of an oncolytic virus. The present invention also provides a drug comprising an activation threshold lowering agent for use in combination with an oncolytic virus to treat cancer, and a drug for treating cancer comprising an activation threshold lowering agent and an oncolytic virus. The present invention further provides the use of an activation threshold lowering agent in the preparation of a drug for treating individual cancer when administered in combination with an oncolytic virus. The present invention further provides the use of an activation threshold lowering agent and an oncolytic virus in the preparation of a drug for treating cancer.

[0470] In some embodiments, the agent that lowers the activation threshold (activation threshold lowering agent) is an agent that reduces the costimulation requirement (costimulation requirement lowering agent) of immune cells (e.g., T cells, B cells and / or NK cells). In some embodiments, the agent that lowers the activation threshold (activation threshold lowering agent) is an agent that promotes tumor immune surveillance. In some embodiments, the agent that lowers the activation threshold (activation threshold lowering agent) is a Cbl-b inhibitor. In some embodiments, the agent that reduces the costimulation requirement is a Cbl-b inhibitor. In some embodiments, the agent that promotes tumor immune surveillance is a Cbl-b inhibitor.

[0471] In some embodiments, activation threshold lowering agents, such as Cbl-b inhibitors, can increase T cell activation and / or T cell proliferation. In some embodiments, activation threshold lowering agents, such as Cbl-b inhibitors, can reduce T cell exhaustion, T cell tolerance and / or T cell anergy.

[0472] In some embodiments, activation threshold lowering agents, such as Cbl-b inhibitors, can increase the generation of one or more cytokines of T cells or peripheral immune cells (e.g., myeloid cells) in the activated T cell microenvironment. In some embodiments, the one or more cytokines include but are not limited to: IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-13, IL-18, TNFα and GM-CSF. In some embodiments, the cytokine is one or more of the following: IL-2, IFN-γ, TNFα and GM-CSF. In some embodiments, the cytokine is a chemokine. In some embodiments, the one or more chemokines include but are not limited to: IP-10, eotaxin (Eotaxin), GROα, RANTES, MIP-1α, MIP-1β, MIP-2, MCP-1 and MCP-3. Increased cytokine expression can be measured by ELISA.

[0473] In some embodiments, activation threshold lowering agents, such as Cbl-b inhibitors, can increase cell surface expression of one or more T cell activation markers. In some embodiments, the one or more T cell activation markers include, but are not limited to, CD25, CD44, CD62L, CD69, CD152 (CTLA4), CD154, CD137, and CD279. In some embodiments, the T cell activation markers are one or more of the following: CD25, CD69, and CTLA4. Increased expression of cell surface markers can be measured by FACS.

[0474] Methods for experimentally determining increased T-cell activation, increased T-cell proliferation, decreased T-cell exhaustion, and / or decreased T-cell tolerance are well known in the art. In some embodiments, representative methods for determining T cell activation can be found in Biological Example 9 and / or Biological Example 13 provided herein. In some embodiments, representative in vitro and in vivo methods for determining increased T cell activation, increased T cell proliferation, decreased T cell exhaustion, and / or decreased T cell tolerance can be found in Biological Example 10 and / or Biological Example 14.

[0475] In some embodiments, activation threshold lowering agents, such as Cbl-b inhibitors, can increase B cell activation. In some embodiments, the increased B cell activation comprises increasing the cell surface expression of one or more B cell activation markers. In some embodiments, the one or more B cell activation markers include but are not limited to: CD69, CD86 and MHC II class (e.g., HLA-DR). In some embodiments, the B cell activation marker is CD69. The expression of cell surface markers increases and can be measured by FACS. In some embodiments, the increased B cell activation comprises protein activation increases in signaling pathways (e.g., those mediated by ERK, JNK and Syk). The increase in protein activation can be detected by measuring the phosphorylation level on the protein using reagents (e.g., anti-phosphorylation antibodies available in the art).

[0476] In some embodiments, activation threshold lowering agents, such as Cbl-b inhibitors, can increase NK cell activation. In some embodiments, the increased NK cell activation comprises the secretion of one or more cytokines. In some embodiments, the one or more cytokines include but are not limited to: IFN-γ, TNFα and MIP-1β. Increased cytokine expression can be measured by ELISA. In some embodiments, the increased NK cell activation comprises increasing the cell surface expression of one or more NK cell activation markers. In some embodiments, the one or more NK cell activation markers include but are not limited to: CD69 and CD107a. Increased expression of cell surface markers can be measured by FACS. In some embodiments, the increased NK cell activation comprises increasing the killing of target cells such as tumor cells (including primary tumor cells) and cell line-derived tumor cells (such as K562 cell line).

[0477] Methods for experimentally determining increased B cell activation and NK cell activation are well known in the art. In some embodiments, representative methods for determining B cell activation can be found in Biological Example 10 and / or Biological Example 14. In some embodiments, representative methods for determining NK cell activation can be found in Biological Example 10 and / or Biological Example 14.

[0478] The activity regulation of immune cells (e.g., T cells, B cells or NK cells) can be measured by measuring the baseline value of the target parameter (e.g., cytokine secretion). For example, T cell activation, such as T cell activation in a sample obtained from an in vitro experiment of cells contacted with a Cbl-b inhibitor, can be measured before contacting or applying the Cbl-b inhibitor to determine a baseline value. A reference value for T cell activation is then obtained after contacting or applying the Cbl-b inhibitor. The reference value is compared with the baseline value to determine the amount of T cell activation caused by contacting or applying the Cbl-b inhibitor or its composition. For example, in some embodiments, compared to the baseline value, the immune cell (e.g., T cell) activation in the sample increases by at least 0.1 times, wherein the baseline value is obtained before the immune cell (e.g., T cell) is contacted with the Cbl-b inhibitor or its composition. In some embodiments, immune cell (for example, T cell) activation increases by at least about 0.1 times, about 0.2 times, about 0.3 times, about 0.4 times, about 0.5 times, about 0.6 times, about 0.7 times, about 0.8 times, about 0.9 times, about 1 times, about 2 times, about 4 times, about 6 times, about 8 times, about 10 times, about 20 times, but no more than about 50 times than baseline value. Immune cell activation can be assessed by measuring biomarkers of activation such as cytokine secretion increase, activation markers (for example, cell surface markers) cell surface expression increase or downstream signaling pathway protein phosphorylation increase. The multiple relative to the baseline value indicating immune cell activation can be determined for the conditions under which the parameters tested and immune cells are treated. For example, in order to measure T cell activation, a baseline value can be obtained from T cells stimulated by a combination of anti-CD3 antibodies and anti-CD28 antibodies, wherein the cells are not incubated with Cbl-b inhibitors. Reference values ​​are then obtained from T cells stimulated with a combination of anti-CD3 and anti-CD28 antibodies, wherein the T cells have been contacted with or are in contact with a Cbl-b inhibitor. A positive response to immune cell activation can then be determined using the reference values ​​obtained. Similar reference value measurements can be obtained and compared to baseline values ​​for assessing T cell activation, T cell proliferation, T cell depletion, T cell tolerance, B cell activation, and / or NK cell activation. Measurements of these parameters can be obtained using techniques well known in the art and the techniques provided in Biological Examples 9, 10, 12, and 13.

[0479] As used herein, the term "baseline" or "baseline value" may refer to a measured value or representation before administering a therapeutic agent disclosed herein (e.g., a composition comprising a Cbl-b inhibitor described herein) or at the start of administration of the therapeutic agent. The baseline value can be compared to a reference value to determine an increase or decrease in immune cell function (e.g., increased T cell activation, increased T cell proliferation, decreased T cell exhaustion, and / or decreased T cell tolerance). As used herein, the term "reference" or "reference value" may refer to a measured value or representation after administering a therapeutic agent disclosed herein (e.g., a composition comprising a Cbl-b inhibitor described herein). The reference value can be measured one or more times during the experimental time course, dosage regimen, or treatment cycle or at the completion of the experimental time course, dosage regimen, or treatment cycle. A "reference value" can be an absolute value, a relative value, a value with an upper and / or lower limit, a range of values, an average value, a median value, an average value, or a value compared to a baseline value. Similarly, a "baseline value" can be an absolute value, a relative value, a value with an upper and / or lower limit, a range of values, an average value, a median value, an average value, or a value compared to a reference value. Reference values ​​and / or baseline values ​​can be obtained from one sample (e.g., one sample obtained from an individual), two different samples (e.g., samples obtained from two different individuals), or a group of samples (e.g., samples obtained from a group of two, three, four, five or more individuals).

[0480] In some embodiments, a positive response to T cell activation as measured by cytokine secretion (e.g., IL-2 secretion) of T cells stimulated with a combination of anti-CD3 and anti-CD28 antibodies in the presence of a Cbl-b inhibitor is at least 2.5 times the baseline value of cytokine secretion (e.g., IL-2 secretion) obtained from T cells stimulated with a combination of anti-CD3 and anti-CD28 antibodies in the absence of a Cbl-b inhibitor. In some embodiments, a positive response to T cell activation as measured by surface marker expression (e.g., CD25 surface marker expression) of T cells stimulated with a combination of anti-CD3 and anti-CD28 antibodies in the presence of a Cbl-b inhibitor is at least 1.3 times the baseline value of surface marker expression (e.g., CD25 surface marker expression) obtained from T cells stimulated with a combination of anti-CD3 and anti-CD28 antibodies in the absence of a Cbl-b inhibitor. In some embodiments, a baseline value can be obtained from T cells stimulated with an anti-CD3 antibody alone, wherein the cells are not incubated with a Cbl-b inhibitor. In some embodiments, a positive response to T cell activation as measured by cytokine secretion (e.g., IL-2 secretion) from T cells stimulated with an anti-CD3 antibody alone in the presence of a Cbl-b inhibitor is at least 0.1 times the baseline value of cytokine secretion (e.g., IL-2 secretion) obtained from T cells stimulated with an anti-CD3 antibody alone in the absence of a Cbl-b inhibitor. In some embodiments, a positive response to T cell activation as measured by surface marker expression (e.g., CD25 surface marker expression) from T cells stimulated with an anti-CD3 antibody alone in the presence of a Cbl-b inhibitor is at least 0.6 times the baseline value of surface marker expression (e.g., CD25 surface marker expression) obtained from T cells stimulated with an anti-CD3 antibody alone in the absence of a Cbl-b inhibitor.

[0481] The present invention provides a method for treating cancer, comprising administering an effective amount of a Cbl-b inhibitor to a subject suffering from cancer, and administering an effective amount of a cancer vaccine to the subject. The present invention also provides a method for treating a subject suffering from cancer, comprising administering an effective amount of a Cbl-b inhibitor to the subject, and administering an effective amount of a cancer vaccine to the subject. Furthermore, the present invention provides a method for increasing an anti-cancer immune response, comprising administering an effective amount of a Cbl-b inhibitor to a subject suffering from cancer, and administering an effective amount of a cancer vaccine to the subject. The present invention further provides a method for treating cancer, comprising administering an effective amount of a Cbl-b inhibitor to a subject suffering from cancer, wherein the subject has received or is currently receiving an effective amount of a cancer vaccine.

[0482] In addition, the present invention provides a method for treating cancer, comprising: administering an effective amount of a Cbl-b inhibitor to an individual suffering from cancer, and administering an effective amount of an oncolytic virus to the individual. The present invention also provides a method for treating an individual suffering from cancer, comprising: administering an effective amount of a Cbl-b inhibitor to the individual; and administering an effective amount of an oncolytic virus to the individual. In addition, the present invention provides a method for increasing an anti-cancer immune response, comprising: administering an effective amount of a Cbl-b inhibitor to an individual suffering from cancer, and administering an effective amount of an oncolytic virus to the individual. The present invention also provides a method for treating cancer, comprising: administering an effective amount of a Cbl-b inhibitor to an individual suffering from cancer, wherein the individual has received or is receiving an effective amount of an oncolytic virus.

[0483] In some embodiments of the methods described in the preceding paragraphs, the Cbl-b inhibitor and the cancer vaccine are administered sequentially in either order. As used herein, the terms "sequentially," "continuously," and "sequentially" refer to administering a Cbl-b inhibitor after administering a cancer vaccine, or administering the cancer vaccine after administering the Cbl-b inhibitor. For example, sequential administration may comprise administering a Cbl-b inhibitor during an induction phase (primary treatment) in the absence of a cancer vaccine, followed by a post-induction treatment phase comprising administering both the cancer vaccine and the Cbl-b inhibitor. The method may further comprise a maintenance phase comprising administering the Cbl-b inhibitor or administering a further dose of the cancer vaccine. Alternatively, sequential administration may comprise administering a cancer vaccine during an induction phase (primary treatment) in the absence of a Cbl-b inhibitor, followed by a post-induction treatment phase comprising administering the Cbl-b inhibitor. The method may further comprise a maintenance phase comprising administering the Cbl-b inhibitor or administering a further dose of the cancer vaccine.

[0484] In some embodiments of the methods described in the aforementioned paragraphs, the Cbl-b inhibitor and the oncolytic virus are administered sequentially in any order. In certain embodiments, as used in the present invention, the terms "sequentially", "continuously" and "sequentially" refer to the administration of a Cbl-b inhibitor after the administration of an oncolytic virus, or the administration of the oncolytic virus after the administration of the Cbl-b inhibitor. For example, sequential administration may include administering a Cbl-b inhibitor during the induction phase (first treatment) in the absence of an oncolytic virus, followed by a post-induction treatment phase comprising administering both an oncolytic virus and a Cbl-b inhibitor. The method may further include a maintenance phase comprising administering the Cbl-b inhibitor or administering a further dose of the oncolytic virus. Alternatively, sequential administration may include administering an oncolytic virus during the induction phase (first treatment) in the absence of a Cbl-b inhibitor, followed by a post-induction treatment phase comprising administering the Cbl-b inhibitor. The method may further include a maintenance phase comprising administering the Cbl-b inhibitor or a further dose of the oncolytic virus.

[0485] In some embodiments of the combination therapy approach, the Cbl-b inhibitor and the cancer vaccine are administered simultaneously. In certain embodiments, as used herein, the terms "simultaneously," "concurrently," and "in parallel" refer to administration of the Cbl-b inhibitor and the cancer vaccine during the same doctor's visit or during the same treatment phase. For example, both the Cbl-b inhibitor and the cancer vaccine can be administered during one or more of the induction phase, the treatment phase, and the maintenance phase. However, simultaneous administration does not require that the Cbl-b inhibitor and the cancer vaccine be present together in a single formulation or pharmaceutical composition, or that the Cbl-b inhibitor and the cancer vaccine be administered at precisely the same time.

[0486] In some embodiments of the combined treatment method, the Cbl-b inhibitor and the oncolytic virus are administered simultaneously. In certain embodiments, as used herein, the terms "simultaneously," "concurrently," and "in parallel" refer to the administration of the Cbl-b inhibitor and the oncolytic virus during the same doctor's visit or during the same treatment phase. For example, both the Cbl-b inhibitor and the oncolytic virus can be administered during one or more of the induction phase, the treatment phase, and the maintenance phase. However, simultaneous administration does not require the Cbl-b inhibitor and the oncolytic virus to be present together in a single formulation or pharmaceutical composition, or does not require the Cbl-b inhibitor and the oncolytic virus to be administered at exactly the same time.

[0487] In some cases, the treatment comprises multiple administrations of a Cbl-b inhibitor or composition thereof, wherein the intervals between administrations can vary. In some embodiments, the Cbl-b inhibitor is administered to the individual at a fixed dose (e.g., mg / adult or mg / child). In some embodiments, the Cbl-b inhibitor is administered to the individual at a fixed dose based on the individual's body weight (e.g., mg / kg).

[0488] In some embodiments, the effectiveness of the combination therapy disclosed herein can be assessed by measuring the biological activity of immune cells present in a sample isolated from the treated individual. For example, immune cells isolated from the individual after treatment can be used to assess the effectiveness of the treatment by measuring their ability to destroy target cells using a cytotoxicity assay. In some embodiments, the biological activity of immune cells present in a sample can be measured by measuring the expression and / or secretion of certain cytokines (e.g., IL-2 and IFNγ).

[0489] Unless otherwise indicated, the term "cancer vaccine" as used herein refers to a "therapeutic cancer vaccine" administered to an individual with cancer for the purpose of treating the cancer (and optionally preventing the recurrence of the cancer). In contrast, a "prophylactic cancer vaccine" (prophylactic cancer vaccine) is administered to an individual who does not have cancer in order to prevent cancer or reduce the risk of the individual developing cancer. Examples of prophylactic cancer vaccines are the human papillomavirus vaccine for the prevention of squamous cell carcinoma and the hepatitis B virus vaccine for the prevention of hepatocellular carcinoma. Cancer vaccines are all immunogenic compositions that include a pharmaceutically acceptable excipient and at least one tumor antigen, such as a tumor-specific antigen or a tumor-associated antigen.

[0490] The terms "oncolytic virus" and "OV" used in the present invention refer to viruses that infect and kill cancer cells. The death of cancer cells is the result of direct cell lysis and anti-tumor immune induction. In some embodiments, an "oncolytic virus" is a replication-competent virus that selectively replicates in cancer cells. In other embodiments, an "oncolytic virus" is a replication-defective virus that does not replicate in cancer cells due to genetic engineering or inactivation (e.g., ultraviolet irradiation or heating) of the oncolytic virus.

[0491] In some embodiments, tumor antigens include "common tumor antigens" shared by many cancers of the same type. Non-limiting examples of shared tumor antigens are breast cancer antigen HER2, prostate cancer antigens PAP and PSA, and melanoma antigens MART-1 and MAGE. In other embodiments, the tumor antigens include "neoantigens" produced as a result of tumor-specific DNA changes (e.g., somatic mutations). Therefore, neoantigens typically have amino acid sequences that are not present in normal mammalian genomes (Schumacher and Schreiber, Science, 348: 69-74, 2015). Non-limiting examples of neoantigens are BRAF V600E, KRAS G12D, KRAS G12V, PIK3CA H1047R, and PIC3CA E545K. The Tumor Specific Neoantigen Database (TSNAdb) is now available free of charge (Wu et al., Genomics Proteomics Bioinformatics 16: 276-282, 2018).

[0492] A variety of techniques are suitable for identifying new antigens contained in cancer vaccines as part of a combination therapy comprising an activation threshold lowering agent (e.g., a Cbl-b inhibitor). For example, new antigens can be identified by the following method, comprising isolating DNA from a tumor biopsy obtained from an individual, sequencing the DNA, and performing computational analysis on the sequence to identify one or more new antigens (Aldous and Dong, Bioorg Med Chem, 26: 2842-2849, 2018). In some embodiments, the computational analysis involves identifying peptides of 8-11 amino acids in length that are expected to bind to at least one HLA allele expressed by tumor cells and contain at least one missense mutation (Wu et al., Genomics Proteomics Bioinformatics 16: 276-282, 2018). New antigens contained in cancer vaccines are thought to be beneficial for overcoming tolerance and reducing autoimmune risks.

[0493] Cancer vaccine platforms suitable for the methods, drugs and uses of the present invention include, but are not limited to, synthetic peptides, recombinant proteins, nucleic acids (DNA or mRNA), microbial vectors, tumor cells and antigen-presenting cells (see, for example, DeMaria and Bilusic, Hematol Oncol Cin North Am, 33: 199-214, 2019; and Maeng and Berzofsky, F1000 Research 2019, 8 (F1000 Faculty Rev): 654, 2019).

[0494] In some embodiments, the tumor antigen of a cancer vaccine comprises at least one synthetic peptide or recombinant protein. In some embodiments, the length of the synthetic peptide is at least 8 amino acids, and in certain embodiments, the length of the synthetic peptide is less than 80 amino acids. In some embodiments, the tumor antigen comprises a plurality of synthetic peptides, or the tumor antigen comprises a synthetic peptide or a recombinant protein, and the synthetic peptide or recombinant protein comprises an amino acid sequence of 2, 3 or more epitopes. An "epitope" is a part of an antigen that binds to an antibody or a B cell receptor, or is presented by a major histocompatibility complex molecule (MHC class I or class II) on the surface of a cell (e.g., a tumor cell or a dendritic cell) for T cell receptor binding. In some embodiments, an epitope is a "linear epitope" (primary structure) consisting of adjacent amino acids of a tumor antigen sequence. In some embodiments, an epitope is a "conformational epitope" (tertiary structure) consisting of non-adjacent amino acids of a tumor antigen. In some embodiments, a tumor antigen comprises a recombinant protein, and the recombinant protein comprises both linear epitopes and conformational epitopes.

[0495] In some embodiments, the tumor antigen is encoded by a DNA or mRNA molecule. In some embodiments, the tumor antigen is encoded by a nucleic acid of a microbial vector, or in other words, the cancer vaccine comprises a microbial vector. In some embodiments, the microbial vector is a live, attenuated microbial vector. In one embodiment, the live, attenuated microbial vector is A live culture preparation of the Bacillus of Calmette and Guerin (BCG) strain of Mycobacterium bovis is sold by Organon USA, Inc. (Roseland, NJ). BCG is approved by the Federal Drug Administration (FDA) for intravesical use after reconstitution with sterile saline (e.g., a pharmaceutically acceptable excipient) for the treatment and prevention of carcinoma in situ of the bladder and for the prevention of primary or recurrent Ta and / or T1 papillary tumors after transurethral resection.

[0496] In some embodiments, the microbial vector is a recombinant microbial vector, such as a recombinant viral vector or a recombinant bacterial vector. The recombinant viral vector suitable for the combination therapy of the present invention includes but is not limited to retrovirus, lentivirus, adenovirus, adeno-associated virus, poxvirus and herpes virus (Chulpanova et al., Biomedicines, 6:94, 2018). In some embodiments, the microbial vector is a recombinant bacterial vector. The recombinant bacterial vector suitable for the combination therapy of the present invention includes but is not limited to Clostridium (Clostridium) (C.novyi), Listeria (Listeria) (for example, Listeria monocytogenes (L.monocytogenes)), Pseudomonas (Pseudomonas) (for example, Pseudomonas aeruginosa (P.aeruginosa)) and Salmonella (Salmonella) (Salmonella typhimurium (S.typhimurium)) (Toussant et al., Expert Rev Vaccines, 12:1139-1154, 2013).

[0497] In some embodiments, the cancer vaccine comprises antigen presenting cells (APCs) that have been contacted with a tumor antigen (e.g., a synthetic peptide or recombinant protein). In some embodiments, the APCs are transfected with a nucleic acid encoding a tumor antigen. In some embodiments, the APCs are transfected with a nucleic acid encoding a cytokine. In some embodiments, the APCs comprise dendritic cells or mesenchymal stem cells. In one embodiment, the cancer vaccine is marketed by Dendreon Corporation (Seattle, WA). (Sipuleucel-T). The invention comprises lactated Ringer cells (eg, a pharmaceutically acceptable excipient) and peripheral blood mononuclear cells (PBMCs) that have been activated with a PAP-GM-CSF fusion protein consisting of prostatic acid phosphatase linked to granulocyte-macrophage colony-stimulating factor. It is an intravenous infusion approved by the Federal Drug Administration (FDA) for the treatment of asymptomatic or minimally symptomatic metastatic prostate cancer.

[0498] In some embodiments, the cancer vaccine comprises killed tumor cells. In some embodiments, the cancer vaccine comprises tumor cell lysate. In some embodiments, the cancer vaccine comprises APCs that have been contacted with tumor cell lysate.

[0499] Adjuvants suitable for use in the cancer vaccines of the methods, medicaments, and uses of the present invention include, but are not limited to, adjuvants of licensed products approved by the FDA. In particular, adjuvants currently approved by the FDA for licensed products include aluminum salts, monophosphoryl lipid A, oil-in-water emulsions (e.g., squalene-in-water emulsions MF59 or AS03), saponins, and CpG oligodeoxynucleotides.

[0500] Oncolytic viruses suitable for use in the methods, medicaments, and uses of the present invention include, but are not limited to, adenovirus, coxsackievirus, echovirus, fowlpox virus, herpes simplex virus, maraba virus, measles virus, myxoma virus, Newcastle disease virus, parvovirus, poliovirus, retrovirus, reovirus, Seneca Valley virus, Semiliki Forest virus, vacciniavirus, and vesicular stomatitis virus (see, e.g., Russell and Peng, Chin Clin Oncol, 7:16, 2018; and Sivanandam et al., Molecular Therapy Oncolytics, 13:93-106).

[0501] In some embodiments, the oncolytic virus is not genetically engineered (non-recombinant virus). In some embodiments, the non-recombinant virus is an echovirus (e.g., Rigvir), Newcastle disease virus, parvovirus, reovirus, or Seneca Valley virus.

[0502] In some embodiments, the oncolytic virus is a recombinant virus that has been genetically engineered to include one or more gene deletions, one or more gene insertions, or one or more gene deletions and one or more gene insertions. In some embodiments, the recombinant virus has been genetically engineered to change the specificity of the host cell and / or the cytotoxicity of the tumor cell. In some embodiments, the recombinant oncolytic virus has been encoded by functional deletions to suppress one or more viral genes of a protein that reacts with the host cell (for example, an antiviral response), and / or by inserting one or more transgenics encoding a protein that promotes the host cell response (for example, an anti-tumor response), genetically engineered (see, for example, Guo et al., Frontiers in Immunology, 8: Article 555, 2017; and Lin et al., Oncology Letters, 15: 4053-4060, 2018). In some embodiments, the recombinant virus is further engineered by inserting a transgenic encoding a detectable marker (such as a fluorescent protein). Ideal anti-tumor response includes one or both of an innate immune response and an adaptive immune response.

[0503] In some embodiments, the recombinant oncolytic virus is a recombinant herpes simplex virus (HSV), such as HSV type 1. In one embodiment, the recombinant oncolytic virus is Also known as talimogene laherparepvec or T-VEC, it is marketed by Amgen Inc. (Thousand Oaks, CA). It is a recombinant HSV-1 that includes functional deletions of the ICP34.5 and ICP47 genes and insertion of nucleic acid encoding human granulocyte macrophage colony-stimulating factor (GM-CSF). It has been approved by the Federal Drug Administration (FDA) for the local treatment of unresectable skin, subcutaneous, and lymph node lesions in patients with recurrent melanoma by intralesional injection (intratumoral administration). It has been approved by the European Medicines Agency (EMA) for the treatment of adult patients with unresectable melanoma (stage IIIB, IIIC, or IVM1a) with regional or distant metastatic disease. Specifically, the EMA-approved product will be administered by intralesional injection (intratumoral administration) into skin, subcutaneous, and / or lymph node lesions that are visible, palpable, or detectable by ultrasound guidance.

[0504] In some embodiments, the recombinant oncolytic virus is a recombinant adenovirus, such as a serotype 5 adenovirus. In one embodiment, the recombinant adenovirus is Oncorine (H101), previously known as Onyx-015. Oncorine is a serotype 5 adenovirus engineered by inactivating viral E1B-55k and viral E3 genes (functional deletion). Oncorine is approved by the China Food and Drug Administration for use in combination with chemotherapy (antitumor therapy) to treat head and neck cancer.

[0505] In some embodiments, the recombinant oncolytic virus is a recombinant poxvirus. In some embodiments, the poxvirus is a vaccinia virus or a fowlpox virus. In some embodiments, the vaccinia virus is a modified Ankara vaccinia virus (Modified Vaccinia Ankara). In some embodiments, the recombinant vaccinia virus has been genetically engineered by functionally deleting one or more viral genes encoding proteins that inhibit host cell responses (e.g., antiviral responses), and / or by inserting one or more transgenes encoding proteins that promote host cell responses (e.g., anti-tumor responses). (See, e.g., Guo et al., Journal of ImmunoTherapy of Cancer, 7:6, 2019). In one embodiment, the recombinant vaccinia virus is Pexa-Vec, also known as pexastimogene devacirepvec and JX-594, which is a vaccinia virus engineered by inactivating (functionally depleting) the viral thymidine kinase gene and inserting transgenes encoding human GM-CSF and β-galactosidase (Heo et al., Nat Med, 19:329-336, 2013). In another embodiment, the recombinant vaccinia virus comprises functionally depleted viral thymidine kinase and vaccinia growth factor genes, and inserting a transgene encoding the chemokine CXCL11 (see, e.g., Liu et al., OncoImmunology, 5:3, e1091554, 2016; and Liu et al., Nature Communications, 8:14754, 2017).

[0506] Further embodiments of the combination therapy of the present invention include at least one additional therapeutic agent. In some embodiments, the at least one additional therapeutic agent is selected from the group consisting of immune checkpoint inhibitors, chemotherapy (anti-tumor agents), radiation therapy, and combinations thereof.

[0507] In some embodiments, the immune checkpoint inhibitor is an antagonist of at least one inhibitory immune checkpoint molecule. In some embodiments, the at least one inhibitory immune checkpoint molecule is selected from the group consisting of PD-1 (CD279), PD-L1 (CD274) and CTLA4 (CD152). The immune checkpoint inhibitor can be a therapeutic biological product. For example, the immune checkpoint inhibitor may comprise an antibody or an antigen-binding fragment thereof. The antibody or fragment may be a monoclonal antibody (mAb), a human antibody, a humanized antibody, or a chimeric antibody, and may include a human constant region. In some embodiments, the human constant region is selected from the group consisting of IgG1, IgG2, IgG3 and IgG4 constant regions, and in certain embodiments, the human constant region is an IgG1 or IgG4 constant region. In some embodiments, the antibody or fragment is a bispecific antibody. In some embodiments, the antigen-binding fragment comprises one of the group consisting of Fab, Fab'-SH, F(ab')2, scFv and Fv fragments.

[0508] In some embodiments, the chemotherapy comprises at least one antineoplastic agent (i.e., WHO ATC code L01). In some embodiments, the at least one antineoplastic agent is selected from the group consisting of: cytotoxic antibiotics, plant alkaloids, antimetabolites, and alkylating agents, other antineoplastic agents, and combinations thereof. In the context of chemotherapy, the antineoplastic agent is a "drug" rather than a "therapeutic biological product."

[0509] In some embodiments, the radiation therapy is external beam radiation therapy. In other embodiments, the radiation therapy is internal beam radiation therapy. In some embodiments, the radiation therapy is ablative radiation therapy.

[0510] The term "biosimilar" or "biosimilar" as used herein refers to a biological product that is similar to a reference product approved by the Federal Drug Administration (FDA) but has no clinically significant differences in safety and efficacy. For example, there may be differences between the biosimilar product and the reference product in terms of clinically inactive ingredients (e.g., differences in formulation excipients, subtle differences in glycosylation, etc.). Clinically significant characteristics can be evaluated through pharmacokinetic and pharmacodynamic studies. In some embodiments, the biosimilar product is an interchangeable product determined by the FDA. In some embodiments, the cancer vaccine is a biosimilar of an FDA-approved product.

[0511] IV. Compositions, Formulations, and Routes of Administration

[0512] The present invention includes pharmaceutical compositions of any compound disclosed herein, or a salt thereof, or a solvate thereof. Thus, the present invention includes pharmaceutical compositions comprising a Cbl-b inhibitor, wherein the Cbl-B inhibitor is a compound of formula (I), (IA)-(IN), (II-A)-(II-L), (III-A)-(III-D), (IV-A)-(IV-C), (VA)-(VG), or (VI-A)-(VI-D) disclosed herein, or any variant thereof, or a pharmaceutically acceptable salt or solvate thereof, or a tautomer thereof, or a stereoisomer thereof, or a mixture of stereoisomers thereof, and a pharmaceutically acceptable excipient, such as a pharmaceutically acceptable vehicle or a pharmaceutically acceptable carrier. In some embodiments, the compound is selected from the group consisting of compounds numbered 1-45 in Table 1, or a pharmaceutically acceptable salt or solvate thereof, or a tautomer thereof, or a stereoisomer thereof, or a mixture of stereoisomers thereof. In some embodiments, the compound is selected from the group consisting of compound numbers 47-52 in Table 1, or a pharmaceutically acceptable salt or solvate thereof, or a tautomer thereof, or a stereoisomer or a mixture of stereoisomers thereof. In one case, the pharmaceutically acceptable salt is an acid addition salt, such as a salt formed with an inorganic acid or an organic acid.

[0513] The compounds and compositions disclosed herein can be administered in any suitable form and by any suitable route to provide sufficient levels of the compound for treating a disease or condition. In some embodiments, the Cbl-b inhibitor and / or the additional therapeutic agent is administered enterally. In some embodiments, the enteral administration is oral administration. In other embodiments, the Cbl-b inhibitor and / or the additional therapeutic agent is administered parenterally. In some embodiments, the parenteral administration is intratumoral injection. In some embodiments, the parenteral administration is by a route selected from the group consisting of intravenous, intraperitoneal, and subcutaneous.

[0514] Suitable routes of administration include oral administration, enteral administration, and parenteral administration includes subcutaneous injection, intravenous injection, intraarterial injection, intramuscular injection, intrasternal injection, intraperitoneal injection, intralesional injection, intraarticular injection, intratumoral injection or infusion technology. The compound and composition can also be administered by sublingual administration, mucosal administration, buccal administration, subcutaneous administration, spinal administration, epidural administration, ventricular administration, suction (for example, as atomizer or spray), nasal administration, vaginal administration, rectal administration, topical administration or transdermal administration, or by sustained release or extended release mechanism. The compound and composition can be administered as needed in the unit dose formulation form containing conventional pharmaceutically acceptable carrier, adjuvant, adjuvant and vehicle. The compound and composition can be directly applied to specific or affected organs or tissues. The compound can be mixed with pharmaceutically acceptable carrier, adjuvant, adjuvant and vehicle to form a composition suitable for required route of administration. In some embodiments, the compound can be mixed with one or both of antigen and adjuvant. In some embodiments, the antigen is a cancer antigen.

[0515] In certain embodiments disclosed herein, particularly those in which the formulation is for injection or other parenteral administration, including the routes listed herein, but also including any other routes of administration described herein (e.g., oral, enteral, gastric, etc.), the formulations and formulations used in these methods are sterile. Sterile pharmaceutical preparations are mixed or formulated according to pharmaceutical grade sterilization standards known to those skilled in the art (U.S. Pharmacopoeia Chapters 797, 1072, and 1211; California Business and Professions Code 4127.7; 16 California Rules and Administrative Rules 1751, 21 U.S. Code of Federal Regulations 211). "Sterile" preparations are sterile, or free of or substantially free of all viable microorganisms and their spores. Examples of sterilization methods for pharmaceutical preparations include, but are not limited to, sterile filtration through a sterile filtration membrane, exposure to radiation (e.g., gamma radiation), and heat sterilization.

[0516] Oral administration is advantageous because of its ease of implementation and patient compliance. If the patient has difficulty swallowing, enteral administration can be accomplished by introducing the drug via a feeding tube, feeding syringe, or gastrostomy. The active compound and other co-administered agents (if present) may be administered enterally in any other pharmaceutically acceptable excipients suitable for administration via a feeding tube, feeding syringe, or gastrostomy.

[0517] Intravenous administration may also be advantageously used to deliver the compound or composition to the bloodstream as rapidly as possible, avoiding the need for absorption from the gastrointestinal tract.

[0518] The compounds and compositions useful for the present invention can be administered in solid form, liquid form, aerosol form, or in the form of tablets, pills, caplets, capsules (e.g., hard gelatin capsules or soft elastic gelatin capsules), powder mixtures, granules, injections, solutions, suppositories, enemas, colon lavages, emulsions, dispersions, food premixes, cachets, lozenges, pastilles, gels, ointments, plasters, pastes, powders, dressings, creams, patches, aerosols (e.g., nasal sprays or inhalers), gels, suspensions (e.g., aqueous or non-aqueous liquid suspensions, oil-in-water emulsions or water-in-oil liquid emulsions), elixirs, or other forms suitable for the administration route. The compounds and compositions can also be administered in liposomal formulations. The compounds can also be administered as prodrugs, wherein the prodrug undergoes conversion to a therapeutically effective form in the subject being treated.

[0519] In addition, the pharmaceutical preparation may include preservatives, solubilizers, stabilizers, rewetting agents, emulsifiers, sweeteners, dyes, regulators, and salts, buffers, coatings, or antioxidants for regulating osmotic pressure. The preparation comprising the compound may also include other substances with valuable therapeutic properties. The pharmaceutical preparation can be prepared by known pharmaceutical methods. Other preparations and methods of administration are known in the art. Suitable preparations can be found in, for example, Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins, 21st ed. (2005), which is incorporated herein by reference.

[0520] Injectable preparations, such as sterile injectable aqueous or oily suspensions, can be prepared using suitable dispersants or wetting agents and suspending agents according to methods known in the art. Sterile injectable preparations can also be sterile injectable solutions or suspensions in parenteral acceptable diluents or solvents, such as propylene glycol solutions. Operable acceptable vehicles and solvents are water, saline, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils can be used as solvents or suspension media generally. In view of this, any gentle fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids, such as oleic acid, can be used in the preparation of injectable preparations.

[0521] Solid dosage forms for oral administration may include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound may be mixed with at least one inert diluent (e.g., sucrose, lactose, talc, or starch). Such dosage forms may also include additional excipients other than the inert diluent, for example, lubricants such as magnesium stearate. In the case of capsules, tablets, and pills, the dosage form may also include a buffer. Tablets and pills may also be prepared with enteric coatings. Acceptable excipients for soft shell gel capsules include, for example, vegetable oils, waxes, fats, semisolid and liquid polyols, and the like.

[0522] Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Such compositions may also contain additional agents such as wetting agents, emulsifiers and suspending agents, cyclodextrins, and sweeteners, flavorings, and fragrances. Alternatively, the compound may be administered in pure form, if appropriate.

[0523] The compounds and compositions can also be administered in the form of liposomes. As is known in the art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by monolayer or multilayer hydrated liquid crystals dispersed in an aqueous medium. Any physiologically acceptable and metabolizable lipid capable of forming liposomes can be used. In addition to the compounds disclosed herein, the compositions of the present invention in liposome form may also contain stabilizers, preservatives, excipients, etc. Useful lipids include natural and synthetic phospholipids and phosphatidylcholines (lecithins). Methods for forming liposomes are all known in the art. See, for example, Gregoriadis, G. Ed., Liposome Technology, Third Edition: Liposome Technology: Liposome Preparation and Related Techniques, CRC Press, Boca Raton, Florida (2006); and Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, New York, NW, p. 33 et seq (1976).

[0524] The amount of active ingredient that can be combined with a carrier material to produce a single dosage form can vary depending on the patient to whom the active ingredient is administered and the specific mode of administration. However, it should be understood that the specific dosage level for any particular patient will depend on a variety of factors, including the specific compound used; the patient's age, weight, body area, body mass index (BMI), general health, sex, and diet; the time and route of administration adopted; the excretion rate; and the drug combination used (if any). The compound can be administered in a unit dose formulation. The selected drug unit dose is prepared and administered to provide a sufficient concentration of the drug in the patient, subject, or individual.

[0525] Although the compounds for use described herein can be administered as the sole active pharmaceutical agent, they can also be used in combination with one or more other pharmaceutical agents. When an additional active agent is used in combination with the compounds for use described herein, the additional active agent can generally be used in therapeutic amounts as set forth in the Physicians' Desk Reference (PDR), 71st edition (2017), which is incorporated herein by reference, or in such therapeutically useful amounts known to those of ordinary skill in the art, or as determined empirically for each patient.

[0526] It is also possible to use a combination of two or more compounds and compositions disclosed herein. The two or more compounds or compositions can be mixed together and administered together shortly before administration. The two or more compounds or compositions can be administered simultaneously by the same route of administration or by different routes of administration. The two or more compounds or compositions can be administered sequentially by the same route of administration or by different routes of administration. In one embodiment, a kit form can include two or more compounds or compositions as a separate form of a compound or composition, with printed or electronic instructions for administering a mixture of compounds or compositions, administering a separate compound or composition simultaneously, or administering a separate compound or composition sequentially. When three or more compounds or compositions are administered, they can be administered as a mixture of compounds or compositions, as a separate compound or composition administered simultaneously, as a separate compound or composition administered sequentially, as a separate compound or composition in which two or more can be administered simultaneously, and the rest can be administered sequentially before or after simultaneous administration, or administered in any other possible combination of mixed administration, simultaneous administration, and sequential administration.

[0527] On the one hand, the compounds disclosed herein can be in purified form, and the present invention discloses compositions comprising the compounds in purified form. The present invention provides compositions comprising the compounds disclosed herein or their salts, such as compositions of substantially pure compounds. In some embodiments, the compositions comprising the compounds disclosed herein or their salts are in substantially pure form. In one variant, "substantially pure" refers to a composition comprising no more than 35% impurities, wherein the impurities represent compounds other than the compound (or multiple compounds, if a combination of compounds is used) administered in the composition, or salts or solvates of this compound (or multiple compounds, if a combination is used). The weight of any added vehicle, carrier, or adjuvant is not included in this calculation, and the added vehicle, carrier, or adjuvant is not considered an impurity. For example, a composition of a substantially pure compound selected from the compounds shown in Table 1 refers to a composition comprising no more than 35% impurities, wherein the impurities represent compounds other than the compound or its salt or solvate. In one variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, wherein the composition comprises no more than 25% impurities. In another variation, the present invention provides a composition of a substantially pure compound, or a salt or solvate thereof, wherein the composition contains no more than 20% impurities. In yet another variation, the present invention provides a composition of a substantially pure compound, or a salt or solvate thereof, wherein the composition contains no more than 10% impurities. In a further variation, the present invention provides a composition of a substantially pure compound, or a salt or solvate thereof, wherein the composition contains no more than 5% impurities. In another variation, the present invention provides a composition of a substantially pure compound, or a salt or solvate thereof, wherein the composition contains no more than 3% impurities. In yet another variation, the present invention provides a composition of a substantially pure compound, or a salt or solvate thereof, wherein the composition contains no more than 1% impurities. In a further variation, the present invention provides a composition of a substantially pure compound, or a salt or solvate thereof, wherein the composition contains no more than 0.5% impurities. In other variations, a composition of a substantially pure compound means that the composition contains no more than 15%, no more than 10%, no more than 5%, no more than 3%, or no more than 1% impurities. The impurity can be a compound in a stereochemical form different from the desired stereochemical form. For example, a composition of a substantially pure (S)-compound means that the composition contains no more than 15%, no more than 10%, no more than 5%, no more than 3%, or no more than 1% of the (R) form of the compound. Alternatively, "enantiomeric excess (ee)" as used herein refers to a dimensionless molar ratio describing the purity of a chiral substance containing, for example, a single stereocenter. For example, an enantiomeric excess of zero would indicate a racemate (e.g., a 50:50 mixture of enantiomers, or no excess of one enantiomer relative to the other).As a further example, an enantiomeric excess of 99 would represent an almost stereopure enantiomer compound (i.e., one enantiomer is in large excess relative to the other). Percent enantiomeric excess, %ee = ([(R)-compound] - [(S)-compound]) / ([(R)-compound] + [(S)-compound]) x 100, where the (R)-compound > (S)-compound; or %ee = ([(S)-compound] - [(R)-compound]) / ([(S)-compound] + [(R)-compound]) x 100, where the (S)-compound > (R)-compound. Additionally, as used herein, "diastereomeric excess (de)" refers to a dimensionless molar ratio describing the purity of a chiral substance containing more than one stereocenter. For example, a diastereomeric excess of zero would represent an equimolar mixture of diastereomers. As a further example, a diastereomeric excess of 99 would represent a nearly stereopure diastereomeric compound (i.e., one diastereoisomer is in large excess relative to the other diastereoisomer). Diastereomeric excess can be calculated by methods similar to ee. As will be appreciated by those skilled in the art, de is typically reported as percent de (% de). % de can be calculated in a manner similar to % ee.

[0528] In some cases, the present invention provides a composition comprising a cell population comprising modified immune cells, such as those described herein or generated by the methods disclosed herein. In some embodiments, the composition comprises a cell population comprising modified immune cells, which have been contacted with or with a Cbl-b inhibitor or a composition thereof described herein. In some embodiments, the modified immune cells have been contacted with or with an anti-CD3 antibody alone. In some embodiments, the modified immune cells have been contacted with or with a combination of an anti-CD28 antibody and an anti-CD3 antibody. The composition provided herein comprising a cell population containing the modified immune cells described herein may further comprise a pharmaceutically acceptable excipient.

[0529] In some cases, the present invention also provides a cell culture composition comprising a cell population comprising the immune cells and Cbl-b inhibitors described herein. In some embodiments, the immune cells are human immune cells. In some embodiments, the immune cells are cells selected from the group consisting of: hematopoietic cells, pluripotent stem cells, myeloid progenitor cells, lymphoid progenitor cells, T cells, B cells, and NK cells. In some embodiments, the cell culture composition further comprises an anti-CD3 antibody. In some embodiments, the cell culture composition further comprises a combination of an anti-CD28 antibody and an anti-CD3 antibody. Methods for culturing cell compositions comprising immune cells are well known in the art and are encompassed by the present invention.

[0530] The modified immune cells or compositions of the present invention, for example, compositions or pharmaceutical compositions comprising a cell population containing the modified immune cells, can be provided in suitable containers. Suitable containers include, for example, bottles, vials (e.g., double-chamber vials), syringes (e.g., single-chamber or double-chamber syringes), bags (e.g., intravenous bags), and tubes (e.g., test tubes). Containers can be formed from a variety of materials (e.g., glass or plastic).

[0531] In some embodiments, a composition (e.g., a cell culture composition) comprising a cell population comprising the modified immune cells of the present invention can be provided in a culture vessel. Culture vessels provided herein include, but are not limited to, tubes (e.g., test tubes), dishes (e.g., tissue culture dishes), bags, multi-well plates (e.g., 6-well tissue culture plates), and flasks (e.g., cell culture flasks).

[0532] The present invention also provides compositions of the present invention for use in any of the uses described herein. In some embodiments, the compositions of the present invention are used in the preparation of a medicament for treating or preventing a disease or condition associated with Cbl-b activity. In some embodiments, the compositions of the present invention are used in the preparation of a medicament for treating cancer.

[0533] The present invention includes pharmaceutical compositions of any compound disclosed herein or a salt or solvate thereof for use in combination with a cancer vaccine. Accordingly, the present invention includes pharmaceutical compositions comprising a Cbl-b inhibitor for use in combination with a cancer vaccine, wherein the Cbl-b inhibitor is a compound of 1-719 (including "a" and "b" variants thereof) of International Patent Application WO 2019 / 148005, or any of Formula (IA), Formula (I), Formula (II-A), Formula (II), Formula (III-A), Formula (III), or Formula (IV), or any variant thereof disclosed therein, or a pharmaceutically acceptable salt or solvate thereof, or a tautomer thereof, or a stereoisomer thereof, or a mixture of stereoisomers thereof.

[0534] In addition, the present invention includes a pharmaceutical composition of any compound disclosed in the present invention used in combination with an oncolytic virus, or its salt or solvate. Therefore, the present invention includes a pharmaceutical composition comprising a Cbl-b inhibitor for use in combination with an oncolytic virus, wherein the Cbl-b inhibitor is a compound of 1-719 (including its "a" and "b" variants) of international patent application WO 2019 / 148005, or formula (IA), formula (I), formula (II-A), formula (II), formula (III-A), formula (III), or formula (IV) Any shown compound, or any variant thereof disclosed therein, or its pharmaceutically acceptable salt or solvate, or its tautomer, or its stereoisomer or mixture of stereoisomers. The composition may further include a pharmaceutically acceptable excipient, such as a pharmaceutically acceptable vehicle or a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition includes both small molecule Cbl-b inhibitors and cancer vaccines. In some embodiments, the compound is a compound selected from the group consisting of compounds numbered 53-61 in Table 1A, or a pharmaceutically acceptable salt or solvate thereof, or a tautomer thereof, or a stereoisomer thereof or a mixture of stereoisomers. In addition, in some embodiments, the pharmaceutical composition comprises both a small molecule Cbl-b inhibitor and an oncolytic virus. In some embodiments, the compound is a compound selected from the group consisting of compounds numbered 53-61 in Table 1A, or a pharmaceutically acceptable salt or solvate thereof, or a tautomer thereof, or a stereoisomer thereof or a mixture of stereoisomers. In some embodiments, the compound is a compound selected from the Cbl-b inhibitors disclosed in the following patent applications: compounds 1-719 (including "a" and "b" variants thereof) of International Patent Application WO 2019 / 148005, or compounds represented by any one of Formula (IA), Formula (I), Formula (II-A), Formula (II), Formula (III-A), Formula (III), or Formula (IV); compounds 1-45 of U.S. Patent Application No. 62 / 831,392 or compounds represented by Formula (I); compounds 1-45 and 47-52 of U.S. Patent Application No. 62 / 866,909 or compounds represented by Formula (I); compounds 1-45 and 47-52 of U.S. Patent Application No. 62 / 880,267 or compounds represented by Formula (I); or any variant thereof, or a pharmaceutically acceptable salt or solvate thereof, or a tautomer thereof, or a stereoisomer or a mixture of stereoisomers thereof.

[0535] In one aspect, the pharmaceutically acceptable salt is an acid addition salt, such as a salt formed with an inorganic acid or an organic acid.

[0536] The compounds, vaccines and compositions disclosed herein can be administered in any suitable form and by any suitable route, which will provide sufficient levels of compounds, vaccines or compositions for treating diseases or conditions. In some embodiments, the Cbl-b inhibitor and / or the cancer vaccine are administered by enteral administration. The compounds, oncolytic viruses and compositions disclosed herein can be administered in any suitable form and by any suitable route, which will provide sufficient levels of compounds, oncolytic viruses or compositions for treating diseases or conditions. In some embodiments, the Cbl-b inhibitor and / or the oncolytic virus are administered by enteral administration. In some embodiments, the enteral administration is oral administration. In other embodiments, the Cbl-b inhibitor and / or the cancer vaccine are administered by parenteral administration. In other embodiments, the Cbl-b inhibitor and / or the oncolytic virus are administered by parenteral administration. In some embodiments, the parenteral administration is intratumoral injection. In some embodiments, the parenteral administration is by a route selected from the group consisting of intravenous, intraperitoneal and subcutaneous.

[0537] Suitable route of administration includes oral administration, enteral administration, and parenteral administration includes subcutaneous injection, intravenous injection, intraarterial injection, intramuscular injection, intrasternal injection, intraperitoneal injection, intralesional injection, intraarticular injection, intratumoral injection or infusion technology. The compound, vaccine and composition can also be administered by sublingual administration, mucosal administration, buccal administration, subcutaneous administration, spinal administration, epidural administration, ventricular administration, suction (for example, as atomizer or spray), nasal administration, vaginal administration, rectal administration, topical administration or transdermal administration, or administered by sustained release or extended release mechanism. The compound, vaccine and composition can be administered as needed with the unit dose formulation form containing conventional pharmaceutically acceptable carrier, adjuvant, adjuvant and vehicle. The compound, vaccine and composition can be directly applied to specific or affected organ or tissue. The compound and / or vaccine can be mixed with pharmaceutically acceptable carrier, adjuvant, adjuvant and vehicle to form a composition suitable for required route of administration. The compound, oncolytic virus and composition can also be administered by sublingual administration, mucosal administration, buccal administration, subcutaneous administration, spinal administration, epidural administration, ventricular administration, inhalation (for example, as an atomizer or spray), nasal administration, vaginal administration, rectal administration, topical administration or transdermal administration, or by sustained release or extended release mechanism. The compound, oncolytic virus and composition can be administered as needed in the form of a unit dose formulation containing conventional pharmaceutically acceptable carriers, excipients, adjuvants and vehicles. The compound, oncolytic virus and composition can be directly applied to specific or affected organs or tissues. The compound and / or oncolytic virus can be mixed with pharmaceutically acceptable carriers, excipients, adjuvants and vehicles to form a composition suitable for the desired route of administration.

[0538] In certain embodiments disclosed in the present invention, especially those embodiments in which the preparation is used for injection or other parenteral administration, including the approach listed in the present invention, but also including any other route of administration described in the present invention (e.g., oral, enteral, gastric, etc.), except for the presence of a microbial carrier in some cancer vaccines, the formulation formula and the preparation used in these methods are sterile. In order to prepare such preparations, all components of the formulation formula or the preparation are prepared in an aseptic manner before being combined with a microbial carrier. In certain embodiments disclosed in the present invention, especially those embodiments in which the preparation is used for injection or other parenteral administration, including the approach listed in the present invention, but also including any other route of administration described in the present invention (e.g., oral, enteral, gastric, etc.), except for the presence of an oncolytic virus, the formulation formula and the preparation used in these methods are sterile. In order to prepare such preparations, all components of the formulation formula or the preparation are prepared in an aseptic manner before being combined with an oncolytic virus. Sterile pharmaceutical preparations are mixed or formulated according to pharmaceutical grade sterilization standards known to those skilled in the art (U.S. Pharmacopoeia Chapters 797, 1072, and 1211; California Business and Professions Code 4127.7; 16 California Rules and Administrative Rules 1751, 21 U.S. Code of Federal Regulations 211). A "sterile" preparation is sterile, or free or substantially free of all viable microorganisms and their spores. Examples of sterilization methods for pharmaceutical preparations include, but are not limited to, sterile filtration through a sterile filtration membrane, exposure to radiation (e.g., gamma radiation), and heat sterilization.

[0539] Oral administration is advantageous because it is easy to implement and patient compliance. If the patient has difficulty swallowing, the drug can be introduced through a feeding tube, a feeding syringe or a gastrostomy to complete enteral administration. Active compound, vaccine or composition and other co-administered medicaments (if present) can be enterally administered in any other pharmaceutically acceptable excipients suitable for the preparation administered via a feeding tube, a feeding syringe or a gastrostomy. Active compound, oncolytic virus or composition and other co-administered medicaments (if present) can be enterally administered in any other pharmaceutically acceptable excipients suitable for the preparation administered via a feeding tube, a feeding syringe or a gastrostomy.

[0540] Intravenous administration may also be advantageously used to deliver the compound, vaccine, or composition to the bloodstream as quickly as possible, avoiding the need for absorption from the gastrointestinal tract. Intravenous administration may also be advantageously used to deliver the compound, oncolytic virus, or composition to the bloodstream as quickly as possible, avoiding the need for absorption from the gastrointestinal tract.

[0541] The compounds, vaccines, and compositions useful for the present invention can be administered in solid form, liquid form, aerosol form, or in the form of tablets, pills, caplets, capsules (e.g., hard gelatin capsules or soft elastic gelatin capsules), powder mixtures, granules, injections, solutions, suppositories, enemas, colon lavages, emulsions, dispersions, food premixes, cachets, troches, lozenges, gels, ointments, plasters, pastes, powders, dressings, creams, patches, aerosols (e.g., nasal sprays or inhalers), gels, suspensions (e.g., aqueous or non-aqueous liquid suspensions, oil-in-water emulsions, or water-in-oil liquid emulsions), elixirs, or other forms suitable for the administration route. The compounds, vaccines, and compositions can also be administered in liposomal formulations. The compounds, oncolytic viruses and compositions that can be used in the present invention can be in solid form, liquid form, aerosol form, or in the form of tablets, pills, caplets, capsules (for example, hard gelatin capsules or soft elastic gelatin capsules), powder mixtures, granules, injections, solutions, suppositories, enemas, colon lavage agents, emulsions, dispersants, food premixes, cachets, lozenges, lozenges, glues, ointments, papules (plasters), pastes, powders, dressings, creams, patches, aerosols (for example, nasal sprays or inhalers), gels, suspensions (for example, aqueous or non-aqueous liquid suspensions, oil-in-water emulsions or water-in-oil liquid emulsions), elixirs, or in the form of other forms suitable for the route of administration. The compounds, oncolytic viruses and compositions can also be administered with liposomal formulations. The compound can also be administered as a prodrug, wherein the prodrug undergoes conversion to a therapeutically effective form in the treated subject.

[0542] In addition, pharmaceutical preparations may include preservatives, solubilizers, stabilizers, rewetting agents, emulsifiers, sweeteners, dyes, regulators, and salts, buffers, coatings, or antioxidants for regulating osmotic pressure. The preparation comprising the compound, vaccine, or composition may also include other substances with valuable therapeutic properties. The preparation comprising the compound, oncolytic virus, or composition may also include other substances with valuable therapeutic properties. Pharmaceutical preparations can be prepared by known pharmaceutical methods. Other preparations and methods of administration are known in the art. Suitable preparations can be found in, for example, Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins, 21st ed. (2005), which is incorporated herein by reference.

[0543] Injectable preparations, such as sterile injectable aqueous or oily suspensions of small molecule Cbl-b inhibitors, can be prepared using suitable dispersants or wetting agents and suspending agents according to methods known in the art. Sterile injectable preparations can also be sterile injectable solutions or suspensions in parenteral acceptable diluents or solvents, such as propylene glycol solutions. Acceptable vehicles and solvents that can be used are water, saline, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils can generally be used as solvents or suspending media. In view of this, any gentle fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids, such as oleic acid, can be used in the preparation of injectable preparations.

[0544] Solid dosage forms for oral administration may include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound, vaccine or composition may be mixed with at least one inert diluent (e.g., sucrose, lactose, talc or starch). In such solid dosage forms, the active compound, oncolytic virus or composition may be mixed with at least one inert diluent (e.g., sucrose, lactose, talc or starch). Such dosage forms may also include additional excipients other than inert diluents, for example, lubricants such as magnesium stearate. In the case of capsules, tablets and pills, the dosage form may also include buffers. Tablets and pills may also be prepared with enteric coatings. Acceptable excipients for soft shell gel capsules include, for example, vegetable oils, waxes, fats, semi-solid and liquid polyols.

[0545] Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Such compositions may also contain additional agents such as wetting agents, emulsifiers and suspending agents, cyclodextrins, and sweeteners, flavorings, and fragrances. Alternatively, the compound may be administered in pure form, if appropriate.

[0546] The compound, vaccine and composition can also be administered in the form of liposomes. The compound, oncolytic virus and composition can also be administered in the form of liposomes. As known in the art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by monolayer or multilayer hydrated liquid crystals dispersed in an aqueous medium. Any physiologically acceptable and metabolizable lipid that can form liposomes can be used. In addition to the compounds or vaccines disclosed in the present invention, the compositions of the present invention in the form of liposomes may also include stabilizers, preservatives, excipients, etc. In addition to the compounds or oncolytic viruses disclosed in the present invention, the compositions of the present invention in the form of liposomes may also include stabilizers, preservatives, excipients, etc. Useful lipids include natural and synthetic phospholipids and phosphatidylcholines (lecithins). Methods for forming liposomes are all known in the art. See, e.g., Gregoriadis, G. Ed., Liposome Technology, Third Edition: Liposome Technology: Liposome Preparation and Related Techniques, CRCPress, Boca Raton, Florida (2006); and Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, New York, NW, p. 33 et seq (1976).

[0547] The amount of active ingredient that can be combined with carrier materials to produce a single dosage form can vary according to the patient and specific mode of administration to which the active ingredient is applied. However, it should be understood that the specific dosage level of any particular patient will depend on a variety of factors, including the specific compound or vaccine used; the patient's age, weight, body area, body mass index (BMI), general health, sex and diet; the administration time and route of administration adopted; excretion rate; and the drug combination used (if any). The compound, vaccine and composition can be administered in unit dose formulations. However, it should be understood that the specific dosage level of any particular patient will depend on a variety of factors, including the specific compound or oncolytic virus used; the patient's age, weight, body area, body mass index (BMI), general health, sex and diet; the administration time and route of administration adopted; excretion rate; and the drug combination used (if any). The compound, oncolytic virus and composition can be administered in unit dose formulations. The selected drug unit dose is prepared and administered to provide a sufficient concentration of drug in a patient, subject or individual.

[0548] Although the compounds, vaccines and compositions for use according to the present invention can be administered as the sole active agent, they can also be used in combination with one or more other agents. When an additional active agent is used in combination with a compound, vaccine or composition for use according to the present invention, the additional active agent can generally be used in a therapeutic amount as shown in the 71st edition (2017) of the Physicians' Desk Reference (PDR), which is incorporated herein by reference, or in such therapeutically useful amounts known to those of ordinary skill in the art, or as determined empirically for each patient. Although the compounds, oncolytic viruses and compositions for use according to the present invention can be administered as the sole active agent, they can also be used in combination with one or more other agents. When an additional active agent is used in combination with the compound, oncolytic virus or composition for use described herein, the additional active agent can generally be used in therapeutic amounts as set forth in the Physicians' Desk Reference (PDR), 71st edition (2017), which is incorporated herein by reference, or in such therapeutically useful amounts known to those of ordinary skill in the art, or as determined empirically for each patient.

[0549] Combinations of two or more compounds, vaccines, and compositions disclosed herein may also be used. The two or more compounds, vaccines, or compositions may be mixed together shortly before administration and administered together. The two or more compounds, vaccines, or compositions may be administered simultaneously by the same route of administration or by different routes of administration. The two or more compounds, vaccines, or compositions may be administered sequentially by the same route of administration or by different routes of administration. In one embodiment, a kit format may include two or more compounds, vaccines, or compositions as separate compounds, vaccines, or compositions, with printed or electronic instructions for administering the compounds, vaccines, or compositions as a mixture, as separate compounds, vaccines, or compositions simultaneously, or as separate compounds, vaccines, or compositions sequentially. When three or more compounds, vaccines, or compositions are administered, they may be administered as a mixture of compounds, vaccines, or compositions, as separate compounds, vaccines, or compositions administered simultaneously, as separate compounds, vaccines, or compositions administered sequentially, as separate compounds, vaccines, or compositions in which two or more may be administered simultaneously and the remaining administered sequentially before or after the simultaneous administration, or as any other possible combination of mixed, simultaneous, and sequential administration.

[0550] A combination of two or more compounds, oncolytic viruses and compositions disclosed in the present invention may also be used. The two or more compounds, oncolytic viruses or compositions may be mixed together and administered together shortly before administration. The two or more compounds, oncolytic viruses or compositions may be administered simultaneously by the same route of administration or by different routes of administration. The two or more compounds, oncolytic viruses or compositions may be administered sequentially by the same route of administration or by different routes of administration. In one embodiment, the kit form may include two or more compounds, oncolytic viruses or compositions as separate forms of a compound, oncolytic virus or composition, with a printed or electronic instruction sheet for administering a mixture of a compound, oncolytic virus or composition, administering a separate compound, oncolytic virus or composition simultaneously, or administering a separate compound, oncolytic virus or composition sequentially. When three or more compounds, oncolytic viruses, or compositions are administered, they can be administered as a mixture of compounds, oncolytic viruses, or compositions, as separate compounds, oncolytic viruses, or compositions that are administered simultaneously, as separate compounds, oncolytic viruses, or compositions that are administered sequentially, as separate compounds, oncolytic viruses, or compositions in which two or more are administered simultaneously and the rest are administered sequentially before or after the simultaneous administration, or in any other possible combination of mixed, simultaneous, and sequential administration.

[0551] The compounds disclosed herein that can be used in the pharmaceutical compositions and methods of the present invention can be in purified form on the one hand, and the present invention discloses compositions comprising the compounds in purified form. The present invention provides compositions comprising the compounds disclosed herein or their salts, such as compositions of substantially pure compounds. In some embodiments, the compositions comprising the compounds disclosed herein or their salts are in substantially pure form. In one variation, "substantially pure" refers to a composition comprising no more than 35% impurities, wherein the impurities represent compounds other than the compound (or multiple compounds, if a combination of compounds is used) administered in the composition, or salts or solvates of the compound (or multiple compounds, if a combination is used). The weight of any added vehicle, carrier, or excipient is not included in this calculation, and the added vehicle, carrier, or excipient is not considered an impurity. For example, a composition of a substantially pure compound selected from the compounds shown in Table 1A refers to a composition comprising no more than 35% impurities, wherein the impurities represent compounds other than the compound or its salt or solvate. In one variation, the present invention provides a composition of a substantially pure compound or its salt or solvate, wherein the composition comprises no more than 25% impurities. In another variation, the present invention provides a composition of a substantially pure compound, or a salt or solvate thereof, wherein the composition contains no more than 20% impurities. In yet another variation, the present invention provides a composition of a substantially pure compound, or a salt or solvate thereof, wherein the composition contains no more than 10% impurities. In a further variation, the present invention provides a composition of a substantially pure compound, or a salt or solvate thereof, wherein the composition contains no more than 5% impurities. In another variation, the present invention provides a composition of a substantially pure compound, or a salt or solvate thereof, wherein the composition contains no more than 3% impurities. In yet another variation, the present invention provides a composition of a substantially pure compound, or a salt or solvate thereof, wherein the composition contains no more than 1% impurities. In a further variation, the present invention provides a composition of a substantially pure compound, or a salt or solvate thereof, wherein the composition contains no more than 0.5% impurities. In other variations, a composition of a substantially pure compound means that the composition contains no more than 15%, no more than 10%, no more than 5%, no more than 3%, or no more than 1% impurities. The impurity can be a compound in a stereochemical form different from the desired stereochemical form. For example, a composition that is substantially pure (S)-compound means that the composition contains no more than 15%, no more than 10%, no more than 5%, no more than 3%, or no more than 1% of the (R) form of the compound.

[0552] Alternatively, as used herein, "enantiomeric excess (ee)" refers to a dimensionless molar ratio describing the purity of a chiral substance containing, for example, a single stereocenter. For example, an enantiomeric excess of zero would represent a racemate (e.g., a 50:50 mixture of enantiomers, or no excess of one enantiomer relative to the other). As a further example, an enantiomeric excess of 99 would represent an almost stereopure enantiomer compound (i.e., a large excess of one enantiomer relative to the other). Percent enantiomeric excess, %ee = ([(R)-compound] - [(S)-compound]) / ([(R)-compound] + [(S)-compound]) x 100, where the (R)-compound > (S)-compound; or %ee = ([(S)-compound] - [(R)-compound]) / ([(S)-compound] + [(R)-compound]) x 100, where the (S)-compound > (R)-compound. Additionally, as used herein, "diastereomeric excess (de)" refers to a dimensionless molar ratio that describes the purity of a chiral substance containing more than one stereocenter. For example, a diastereomeric excess of zero would represent an equimolar mixture of diastereomers. As a further example, a diastereomeric excess of 99 would represent a nearly stereopure diastereomeric compound (i.e., one diastereomer is in large excess relative to the other). Diastereomeric excess can be calculated in a similar manner to ee. As will be appreciated by those skilled in the art, de is typically reported as percent de (% de). % de can be calculated in a similar manner to % ee.

[0553] The compound, vaccine or composition disclosed in the present invention can be provided in a suitable container. The compound, oncolytic virus or composition disclosed in the present invention can be provided in a suitable container. Suitable containers include, for example, bottles, vials (e.g., double-chamber vials), syringes (e.g., single-chamber or double-chamber syringes), bags (e.g., intravenous bags) and tubes (e.g., test tubes). The container can be formed of a variety of materials (e.g., glass or plastic).

[0554] V. Products or Kits

[0555] The present invention also provides articles comprising any of the compounds, pharmaceutical compositions, cells, modified immune cells, cell populations, cell compositions, cell cultures, or cell culture compositions described herein. The articles include suitable containers or packaging materials for the compounds, pharmaceutical compositions, cells, modified immune cells, cell populations, cell compositions, cell cultures, or cell culture compositions. Examples of suitable containers include, but are not limited to, bottles, vials, syringes, intravenous bags, or tubes. For cells, modified immune cells, cell populations, cell compositions, cell cultures, or cell culture compositions, suitable containers can be culture vessels, including, but not limited to, tubes, dishes, bags, multiwell plates, or flasks.

[0556] The present invention also provides a kit comprising any compound, pharmaceutical composition, cell, modified immune cell, cell group, cell composition, cell culture, or cell culture composition described herein. The kit may include a compound, pharmaceutical composition, cell, modified immune cell, cell group, cell composition, cell culture, or cell culture composition in a suitable container or packaging material, and the suitable container or packaging material may include but is not limited to a bottle, vial, syringe, intravenous bag, or tube. The kit may include a cell, modified immune cell, cell group, cell composition, cell culture, or cell culture composition in a culture vessel, and the culture vessel may include but is not limited to a tube, dish, bag, multi-well plate, or flask. The kit may include the compound, pharmaceutical composition, cell, modified immune cell, cell group, cell composition, cell culture, or cell culture composition for administration to an individual in a single dose or multiple dose form. The kit may further include instructions or labels for administering the compound, pharmaceutical composition, cell, modified immune cell, cell group, cell composition, cell culture, or cell culture composition to an individual according to any method disclosed herein. The kit may further comprise a device for administering the compound, pharmaceutical composition, cell, modified immune cell, cell population, cell composition, cell culture, or cell culture composition to an individual, including but not limited to a needle, syringe, catheter, or intravenous bag. The kit may further comprise instructions for producing any compound, pharmaceutical composition, cell, modified immune cell, cell population, cell composition, cell culture, or cell culture composition disclosed herein.

[0557] The present invention also provides a product comprising any compound, vaccine, or pharmaceutical composition described herein. The product includes a suitable container or packaging material for the compound, vaccine, or pharmaceutical composition. The present invention also provides a product comprising any compound, oncolytic virus, or pharmaceutical composition described herein. The product includes a suitable container or packaging material for the compound, oncolytic virus, or pharmaceutical composition. Examples of suitable containers include, but are not limited to, bottles, vials, syringes, intravenous bags, or tubes.

[0558] The present invention also provides a kit comprising any compound, vaccine or pharmaceutical composition described herein. The kit may include the compound, vaccine or pharmaceutical composition in an appropriate container or packaging material, including but not limited to bottles, vials, syringes, intravenous bags or tubes. The kit may include the compound, vaccine or pharmaceutical composition for administration to an individual in a single dose form or a multi-dose form. The kit may further include instructions or labels for administering the compound, vaccine or pharmaceutical composition to an individual according to any method disclosed in the present invention. The kit may further include equipment for administering the compound, vaccine or pharmaceutical composition to an individual, including but not limited to a needle, syringe, catheter or intravenous bag. The kit may further include instructions for generating any compound, vaccine or pharmaceutical composition disclosed in the present invention.

[0559] The present invention also provides a kit comprising any compound, oncolytic virus or pharmaceutical composition described in the present invention. The kit may include the compound, oncolytic virus or pharmaceutical composition in a suitable container or packaging material, and the suitable container or packaging material may include but is not limited to bottles, vials, syringes, intravenous infusion bags or tubes. The kit may include the compound, oncolytic virus or pharmaceutical composition for application to an individual in a single dose form or a multi-dose form. The kit may further include instructions or labels for administering the compound, oncolytic virus or pharmaceutical composition to an individual according to any method disclosed in the present invention. The kit may further include equipment for administering the compound, oncolytic virus or pharmaceutical composition to an individual, including but not limited to a needle, a syringe, a catheter or an intravenous infusion bag. The kit may further include instructions for generating any compound, oncolytic virus or pharmaceutical composition disclosed in the present invention.

[0560] VI. Exemplary Embodiments

[0561] The present invention is further described by the following embodiments. Where appropriate and practical, the features of each embodiment can be combined with any other embodiment.

[0562] Embodiment 1. Compound represented by formula (Ia):

[0563]

[0564] or a tautomer thereof, or a pharmaceutically acceptable salt thereof,

[0565] in:

[0566] yes

[0567] X is CH or N;

[0568] Z 1 is CH or N;

[0569] Z 2 is CH or N;

[0570] R 1a and R 1b are independently H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH;

[0571] R 2a is -CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyl-OH, C1-C6 alkyl-CN, or -(C1-C6 alkylene)-O-(C1-C6 alkyl);

[0572] R 2b is H, halogen, or C1-C6 alkyl;

[0573] or R 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro heterocyclic group consisting of 3 to 6 atoms, or a spiro C3-C6 cycloalkyl group, wherein at least one of the atoms of the spiro heterocyclic group adjacent to the piperidinyl ring to which it is attached is carbon;

[0574] R 3a and R 3b are independently H, halogen, or C1-C6 alkyl;

[0575] or R 3a and R 3b Together with the carbon atom to which it is attached, it forms a C3-C4 cycloalkyl group;

[0576] R 4 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH;

[0577] yes and Y is CR 5a R 5b or S;

[0578] yes and Y are bonds;

[0579] W is O or a bond;

[0580] R 5a and R 5b are independently H, halogen, or C1-C6 alkyl;

[0581] or R 5a and R 5b Together with the carbon atom to which it is attached, it forms a C3-C6 cycloalkyl group;

[0582] R 6 is a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C6 cycloalkyl group;

[0583] R 7 is H, C3-C6 cycloalkyl, -NH-(heterocyclyl consisting of 3 to 6 atoms), -NH-(C1-C6 alkyl), -NH-(C3-C6 cycloalkyl), -O-(heterocyclyl consisting of 3 to 6 atoms), -O-(C1-C6 alkyl), or -O-(C3-C6 cycloalkyl);

[0584] R 8 is a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C6 cycloalkyl group;

[0585] R 9 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH; and

[0586] R 10 It is -CF3 or cyclopropyl.

[0587] Embodiment 2. The compound according to embodiment 1, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0588] yes

[0589] Embodiment 3. The compound according to embodiment 2, or its tautomer, or its pharmaceutically acceptable salt, wherein: Z 1 It is CH.

[0590] Embodiment 4. The compound according to Embodiment 3, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0591] yes

[0592] Embodiment 5. The compound according to embodiment 2, or its tautomer, or its pharmaceutically acceptable salt, wherein: Z 1 It's N.

[0593] Embodiment 6. The compound according to Embodiment 5, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0594] yes

[0595] Embodiment 7. The compound according to Embodiment 1, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0596] yes

[0597] Embodiment 8. The compound according to embodiment 7, or its tautomer, or its pharmaceutically acceptable salt, wherein: Z 2 It is CH.

[0598] Embodiment 9. The compound according to embodiment 8, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0599] yes

[0600] Embodiment 10. The compound according to embodiment 7, or its tautomer, or its pharmaceutically acceptable salt, wherein: Z 2 It's N.

[0601] Embodiment 11. The compound according to Embodiment 10, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0602] yes

[0603] Embodiment 12. The compound according to any one of Embodiments 1-11, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0604] yes and Y is CR 5a R 5b or S.

[0605] Embodiment 13. The compound according to Embodiment 12, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0606] W is a key.

[0607] Embodiment 14. The compound according to Embodiment 12, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0608] W is O.

[0609] Embodiment 15. The compound according to any one of Embodiments 1-11, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0610] yes and Y are bonds.

[0611] Embodiment 16. The compound according to Embodiment 15, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0612] R 8 It is a C1-C3 alkyl group, a C1-C3 haloalkyl group, or a C3-C4 cycloalkyl group.

[0613] Embodiment 17. The compound according to Embodiment 16, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0614] R 8 It is -CH3, -CF3, or cyclopropyl.

[0615] Embodiment 18. The compound according to any one of Embodiments 1-17, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0616] X is CH.

[0617] Embodiment 19. The compound according to any one of Embodiments 1-17, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0618] X is N.

[0619] Embodiment 20. The compound according to any one of Embodiments 1-19, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0620] Y is a bond.

[0621] Embodiment 21. The compound according to any one of Embodiments 1-19, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0622] Y is CR 5a R 5b .

[0623] Embodiment 22. The compound according to Embodiment 21, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0624] R 5a and R 5b are independently H, halogen, or C1-C3 alkyl;

[0625] or R 5a and R 5b Together with the carbon atom to which it is attached, it forms a C3-C4 cycloalkyl group.

[0626] Embodiment 23. The compound according to Embodiment 22, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0627] R 5a and R 5b are independently H, F, or -CH3;

[0628] or R 5a and R 5b Together with the carbon atom to which it is attached, it forms a cyclopropyl group.

[0629] Embodiment 24. The compound according to Embodiment 23, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0630] R 5a and R 5b are independently H or F.

[0631] Embodiment 25. The compound according to any one of Embodiments 1-19, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0632] Y is S.

[0633] Embodiment 26. The compound according to any one of Embodiments 1-25, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0634] R 1a and R 1b are independently H, C1-C3 alkyl, C1-C3 haloalkyl, or C1-C3 alkyl-OH.

[0635] Embodiment 27. The compound according to Embodiment 26, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0636] R 1a and R 1b are independently H, -CH3, -CF3, or -CH2OH.

[0637] Embodiment 28. The compound according to any one of Embodiments 1-27, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0638] R 1b It’s H.

[0639] Embodiment 29. The compound according to any one of Embodiments 1-28, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0640] R 2a It is -CN, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyl-OH, C1-C3 alkyl-CN, or -(C1-C3 alkylene)-O-(C1-C3 alkyl).

[0641] Embodiment 30. The compound of Embodiment 29, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0642] R 2a It is -CN, -CH3, -CF3, -CH2OH, -CH2CN, or -CH2-O-CH3.

[0643] Embodiment 31. The compound according to any one of Embodiments 1-30, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0644] R 2b is H, h...

Claims

1. Compounds of formula (I) or a pharmaceutically acceptable salt thereof, wherein yes or X is CH or N; Z 1 is CH or N; Z 2 is CH or N; R 1a and R 1b are independently H, C1-C6 alkyl or C1-C6 haloalkyl; R 2a is -CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyl-OH or C1-C6 alkyl-CN; R 2b is H, halogen, or C1-C6 alkyl; or R 2a and R 2b together with the carbon atom to which it is attached, forms a spiro 3- to 6-membered heterocyclyl or a spiro C3-C6 cycloalkyl, wherein at least one of the atoms of the spiro heterocyclyl adjacent to the piperidinyl ring to which it is attached is carbon; R 3a and R 3b are independently H or halogen; R 4 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH; yes and Y is CR 5a R 5b or sulfur; or yes and Y are bonds; W is oxygen or a bond; R 5a and R 5b are independently H, halogen, or C1-C6 alkyl; or R 5a and R 5b Together with the carbon atom to which it is attached, it forms a C3-C6 cycloalkyl group; R 6 is C1-C6 alkyl, C1-C6 haloalkyl, or C3-C6 cycloalkyl; R 7 is H, halogen, C3-C6 cycloalkyl, -NH-(3- to 6-membered heterocyclyl), -NH-(C1-C6 alkyl), -NH-(C3-C6 cycloalkyl), -O-(3- to 6-membered heterocyclyl), -O-(C1-C6 alkyl), or -O-(C3-C6 cycloalkyl); R 8 is C1-C6 alkyl, C1-C6 haloalkyl, or C3-C6 cycloalkyl; R 9 is H or C1-C6 alkyl; and R 10 It is -CF3 or cyclopropyl.

2. The compound according to claim 1, wherein yes 3. The compound according to claim 2, wherein Z 1 It is CH; Optionally yes or or Z 1 is N; Optionally yes or 4. The compound according to claim 1, wherein yes 5. The compound according to claim 4, wherein Z 2 It is CH; Optionally yes or or Z 2 is N; Optionally yes or 6. The compound according to claim 1, wherein yes and Y is CR 5a R 5b or sulfur; optionally wherein W is a bond; or W is oxygen; or in yes and Y are bonds.

7. The compound according to claim 6, wherein R 8 It is a C1-C3 alkyl group, a C1-C3 haloalkyl group, or a C3-C4 cycloalkyl group.

8. The compound according to claim 7, wherein R 8 It is -CH3, -CF3, or cyclopropyl.

9. The compound according to claim 1, wherein X is CH; or X is N; and / or wherein Y is a bond; Y is CR 5a R 5b ;or Y is sulfur.

10. The compound according to claim 9, wherein Y is CR 5a R 5b and R 5a and R 5b are independently H, halogen, or C1-C3 alkyl, or R 5a and R 5b Together with the carbon atom to which it is attached, it forms a C3-C4 cycloalkyl group.

11. The compound according to claim 10, wherein R 5a and R 5b are independently H, F, or -CH3; or R 5a and R 5b Together with the carbon atom to which it is attached it forms a cyclopropyl group.

12. The compound according to claim 11, wherein R 5a and R 5b are independently H or F.

13. The compound according to claim 1, wherein R 1a and R 1b are independently H, C1-C3 alkyl or C1-C3 haloalkyl.

14. The compound according to claim 13, wherein R 1a and R 1b are independently H, -CH3 or -CF3.

15. The compound according to claim 14, wherein R 1b It's H.

16. The compound according to claim 1, wherein R 2a It is -CN, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyl-OH or C1-C3 alkyl-CN.

17. The compound according to claim 16, wherein R 2a is -CN, -CH3, -CF3, -CH2OH or -CH2CN; and / or Where R 2b is H, halogen or C1-C3 alkyl.

18. The compound according to claim 17, wherein R 2b It is H, F, or -CH3.

19. The compound according to claim 1, wherein R 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro 4- to 5-membered heterocyclyl, or a spiro C3-C4 cycloalkyl.

20. The compound according to claim 19, wherein R 2a and R 2b together with the carbon atom to which it is attached, forms a spirocyclopropyl group, or 21. The compound according to claim 1, wherein R 3a and R 3b are independently H or F.

22. The compound according to claim 1, wherein R 4 It is H, C1-C3 alkyl, C1-C3 haloalkyl, or C1-C3 alkyl-OH.

23. The compound according to claim 22, wherein R 4 is H, -CH3, -CF3, or -CH2OH; and / or Where R 6 It is a C1-C3 alkyl group, a C1-C3 haloalkyl group, or a C3-C4 cycloalkyl group.

24. The compound according to claim 23, R 6 It is -CH3, -CHF2, or cyclopropyl.

25. The compound according to claim 1, wherein R 7 It is H, halogen, C3-C4 cycloalkyl, -NH-(4- to 5-membered heterocyclyl), -NH-(C1-C3 alkyl), -NH-(C3-C5 cycloalkyl), -O-(C1-C3 alkyl), -O-(4- to 5-membered heterocyclyl), or -O-(C3-C5 cycloalkyl).

26. The compound according to claim 25, wherein R 7 is H, Cl, cyclopropyl, -NH(CH2CH3), -NH(cyclopropyl), -OCH2CH3, -O-(cyclopropyl), or and / or Where R 9 It is H or C1-C3 alkyl.

27. The compound according to claim 26, wherein R 9 It is H or -CH3.

28. The compound according to claim 1, wherein the compound is a compound of formula (Ia): or a pharmaceutically acceptable salt thereof, wherein: yes or X is CH or N; Z 1 is CH or N; Z 2 is CH or N; R 1a and R 1b are independently H, C1-C6 alkyl or C1-C6 haloalkyl; R 2a is -CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyl-OH or C1-C6 alkyl-CN; R 2b is H, halogen, or C1-C6 alkyl; or R 2a and R 2b Together with the carbon atom to which it is attached, it forms a spiro 3- to 6-membered heterocyclyl, or a spiro C3-C6 cycloalkyl, wherein at least one of the atoms of the spiro heterocyclyl adjacent to the piperidinyl ring to which it is attached is carbon; R 3a and R 3b are independently H or halogen; R 4 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkyl-OH; yes and Y is CR 5a R 5b or sulfur; or yes and Y are bonds; W is oxygen or a bond; R 5a and R 5b are independently H, halogen, or C1-C6 alkyl; or R 5a and R 5b Together with the carbon atom to which it is attached, it forms a C3-C6 cycloalkyl group; R 6 is C1-C6 alkyl, C1-C6 haloalkyl, or C3-C6 cycloalkyl; R 7 is H, C3-C6 cycloalkyl, -NH-(3- to 6-membered heterocyclyl), -NH-(C1-C6 alkyl), -NH-(C3-C6 cycloalkyl), -O-(3- to 6-membered heterocyclyl), -O-(C1-C6 alkyl), or -O-(C3-C6 cycloalkyl); R 8 is C1-C6 alkyl, C1-C6 haloalkyl, or C3-C6 cycloalkyl; R 9 is H or C1-C6 alkyl; and R 10 It is -CF3, or cyclopropyl.

29. The compound according to claim 1, selected from 30. The compound according to claim 1, selected from 31. A pharmaceutical composition comprising the compound of claim 1 and a pharmaceutically acceptable excipient.

32. Use of a Cbl-b inhibitor in the manufacture of a medicament for regulating the activity of immune cells, wherein the immune cells are T cells, wherein the Cbl-b inhibitor is the compound of claim 1.

33. The use according to claim 32, wherein the immune cells have been isolated or are isolated from a blood sample of a mammalian subject; or wherein the immune cell is a tumor infiltrating lymphocyte (TIL) that has been isolated or is isolated from a tumor of a mammalian subject suffering from cancer; or wherein regulating the T cell activity comprises one or more of increased T cell activation, increased T cell proliferation, decreased T cell exhaustion, and decreased T cell tolerance; or The immune cells are human immune cells; or The immune cell comprises a recombinant chimeric receptor.

34. The use of claim 33, wherein the increased T cell activation comprises increased production of cytokines; or wherein the increased T cell activation comprises increased cell surface expression of one or more T cell activation markers.

35. The use according to claim 34, wherein the cytokine comprises one or more selected from the group consisting of IL-2, IFN-γ, TNFα and GM-CSF.

36. The use according to claim 34, wherein the T cell activation marker comprises one or more selected from the group consisting of CD25, CD69 and CTLA4.

37. The use according to claim 33, wherein the T cells have been contacted with anti-CD3 antibodies alone or in combination with anti-CD28 antibodies, or the T cells have been contacted with anti-CD3 antibodies alone or in combination with anti-CD28 antibodies.

38. The use according to claim 33, further comprising culturing the immune cells with IL-2 alone or in combination with anti-CD3 antibody and / or anti-CD28 antibody.

39. The use according to claim 33, wherein the recombinant chimeric receptor is a chimeric antigen receptor.

40. A method for generating an expanded tumor infiltrating lymphocyte (TIL) population ex vivo for non-therapeutic purposes, the method comprising: (a) obtaining a biological sample comprising TILs from an individual suffering from cancer who has received or is receiving an effective amount of a Cbl-b inhibitor, wherein the Cbl-b inhibitor is a compound according to claim 1; and (b) culturing the TILs in a cell culture medium comprising at least one T cell growth factor to generate an expanded TIL population.

41. The method of claim 40, wherein the at least one T cell growth factor comprises IL-2.

42. The method of claim 40, wherein the cell culture medium further comprises an anti-CD3 antibody, or both an anti-CD3 antibody and an anti-CD28 antibody; or The cell culture medium further comprises the Cbl-b inhibitor.

43. The method of claim 40, wherein the cancer is a non-hematological cancer, optionally wherein the non-hematological cancer is a sarcoma, carcinoma, or melanoma.

44. A composition comprising the expanded TIL population generated by the method of claim 40, and a physiologically acceptable buffer.

45. Use of the composition of claim 44 in the manufacture of a medicament for treating cancer.

46. ​​The use according to claim 45, further comprising continuing to administer an effective amount of the Cbl-b inhibitor.

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