Substituted benzyl-triazole compounds for Cbl-b inhibition and their further uses

Abstract

The present invention discloses compounds, compositions and methods of formula (I) for inhibiting the E3 enzyme Cbl-b in the ubiquitin proteasome pathway. The compounds, compositions and methods can be used to modulate the immune system for treating diseases suitable for immune system modulation, and for in vivo, in vitro or ex vivo cell therapy. 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 the following U.S. Provisional Application Nos. under 35 U.S.C. § 119: 62 / 866,914, filed Jun. 26, 2019; 62 / 880,285, filed Jul. 30, 2019; 62 / 888,845, filed Aug. 19, 2019; and 62 / 888,870, filed Aug. 19, 2019, the entire contents of each of which are incorporated herein by reference in their entirety. Field of the Invention

[0003] The present invention provides compounds and compositions for inhibiting the Cbl - b enzyme and methods of using the same 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 of treating cancer comprising a combination of the Cbl - b enzyme inhibitor and a cancer vaccine; and pharmaceutical compositions, kits, and methods of treating cancer comprising a combination of the Cbl - b enzyme inhibitor and an oncolytic virus. Background of the Invention

[0005] The ubiquitin - proteasome pathway is a complex system involved in the regulation of protein function and catabolism. In eukaryotic cells, proteins are conjugated to ubiquitin, a 76 - amino - acid, 8.5 - kilodalton protein. This conjugation, known as ubiquitination, results in either a change in the function of the target protein or its degradation. Ubiquitination of the target protein occurs through a series of coupled reactions involving ubiquitin and a group of enzymes called E1, E2, and E3 enzymes. Ubiquitin is activated by a ubiquitin - activating enzyme or E1 enzyme. Ubiquitin is then transferred to a ubiquitin - conjugating enzyme or E2 enzyme. Finally, a ubiquitin - ligase or E3 enzyme facilitates the transfer of ubiquitin from the E2 enzyme to the target protein. Poly - ubiquitination of the target protein mainly 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 a variety of biological processes, including cell division, DNA repair, and cell signaling.

[0006] The synthesis and degradation of proteins in cells are crucial for cell - cycle regulation, cell proliferation, apoptosis, and many other cellular processes. Thus, the ability to regulate the ubiquitin - proteasome pathway provides numerous opportunities for intervening in disease processes. Intervention mechanisms can include enhancing the degradation of oncogene products, reducing the degradation of tumor - suppressor proteins, modulating immune - cell responses, and modulating anti - tumor immune responses.

[0007] Therapeutic cancer vaccines have been evaluated in many clinical trials. However, only two therapeutic cancer vaccines are approved for use in the United States. In particular, the Bacillus of Calmette and Guerin 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. Even so, the response rates and overall survival rates of patients receiving cancer vaccine treatment are still far below ideal levels. Accordingly, what is needed in the art are methods for improving the efficacy of cancer vaccines.

[0008] Although numerous clinical trials have been conducted using oncolytic viruses to treat cancer, only one oncolytic virus is approved for use in the United States and Europe. In particular, talimogene laherparepvec is a genetically modified herpes simplex virus that 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. Accordingly, what is needed in the art are methods for improving the efficacy of oncolytic virus therapies.

[0009] Approximately 35 E2 enzymes and over 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, 2012:692639). Agents that modulate E2 or E3 enzymes correspondingly offer potential as therapies for 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 for use in combination with cancer vaccines, and pharmaceutical compositions comprising a Cbl-b inhibitor and a cancer vaccine; and agents that inhibit Cbl-b for use in combination with oncolytic viruses, and pharmaceutical compositions comprising a Cbl-b inhibitor and an oncolytic virus. Abstract of the Invention

[0010] The present invention discloses compounds and compositions for inhibiting the Cbl-b enzyme and methods of using them to modulate the immune system, treat diseases, and treat cells in vivo, in vitro, or ex vivo. The present invention also discloses methods of using a Cbl-b inhibitor to treat cancer. Briefly, the Cbl-b inhibitor can be administered alone to an individual with cancer or as part of a combination therapy together with one or more of immune checkpoint inhibitors, anti-tumor agents, and radiotherapy to an individual with cancer. In addition, cells treated in vivo and / or in vitro with the compounds or compositions disclosed in the present invention can be used for adoptive cell therapy for treating cancer.

[0011] The present invention discloses a compound represented by formula (I):

[0012]

[0013] or a tautomer thereof, or a pharmaceutically acceptable salt of any of the foregoing substances,

[0014] wherein:

[0015]

[0016] Z 1 is CH or N;

[0017] Z 2 is CH or N;

[0018] R 1 is -CF 3 or cyclopropyl;

[0019] R 2 is -CF 3 or cyclopropyl;

[0020] R 3 is H, C 1 -C 2 alkyl, or C 1 -C 2 haloalkyl;

[0021] R 4 is H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, a heterocyclic group composed of 4 to 8 atoms, or C 3 -C 6 cycloalkyl,

[0022] wherein the heterocyclic group or cycloalkyl group is each optionally substituted with 1 - 5 R 6 groups;

[0023] or R 3 and R 4 together with the carbon atom to which they are attached form a C 3 -C 5 cycloalkyl, or a heterocyclic group composed of 4 to 6 atoms, wherein C 3 -C 5 cycloalkyl and the heterocyclic group composed of 4 to 6 atoms are each optionally substituted with 1 - 5 R 6 groups;

[0024] R 5 is H, C 1 -C 6alkyl, C 1 -C 6 haloalkyl, or C 3 -C 6 cycloalkyl;

[0025] Each R 6 is independently C 1 -C 6 alkyl, halogen, hydroxy, -O(C 1 -C 6 alkyl), -CN, C 1 -C 6 alkyl-CN, C 1 -C 6 alkyl-OH, or C 1 -C 6 haloalkyl;

[0026] Or two R 6 groups together with the carbon atom to which they are attached form a spiro C 3 -C 6 cycloalkyl, or a heterocyclic group consisting of 4 to 6 atoms in a spiro;

[0027] X is H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkyl-OH, C 1 -C 6 alkyl-CN, optionally substituted by 1 to 5 R 8 groups, C 3 -C 6 cycloalkyl, or

[0028] is a heterocyclic group consisting of 4 to 7 atoms, or a heteroaryl group consisting of 5 to 8 atoms, where the heterocyclic group or heteroaryl group each optionally contains 1 to 2 additional heteroatoms selected from the group consisting of N, O, and S, and where the heterocyclic group or heteroaryl group each optionally is substituted by 1 to 5 R 8 groups;

[0029] Each R 7 is independently H, C 1 -C 6 alkyl, C 1 -C 6 alkyl-OH, or C 1 -C 6 haloalkyl;

[0030] Or two R 7The group together with the carbon atom to which it is attached forms a C 3 -C 5 cycloalkyl group, or a heterocyclic group composed of 3 to 5 atoms; and each R 8 is independently halogen, C 1 -C 6 alkyl, C 1 -C 6 alkyl-CN, C 1 -C 6 alkyl-OH, C 1 -C 6 haloalkyl, -CN, oxo,

[0031] or -O(C 1 -C 6 alkyl);

[0032] or two R 8 groups together with the carbon atom to which they are attached form a spiro or fused C 3 -C 5 cycloalkyl group, or a spiro or fused heterocyclic group composed of 3 to 5 atoms.

[0033] In some embodiments, In some embodiments, Z 1 is CH. In some embodiments, Z 1 is N.

[0034] In some embodiments,

[0035] In some embodiments, In some embodiments, Z 2 is CH. In some embodiments, Z 2 is N.

[0036] In some embodiments,

[0037]

[0038] In some embodiments, R 3 is H, C 1 -C 2 alkyl, or C 1 -C 2 haloalkyl; R 4 is H, C 1 -C 3 alkyl, C 1 -C 6 haloalkyl, a heterocyclic group composed of 4 to 6 atoms, or C 4 -C 5A cycloalkyl group, wherein the heterocyclic group or cycloalkyl group is each optionally substituted with 1-3 R 6 groups; or R 3 and R 4 together with the carbon atom to which they are attached form a C 4 -C 5 cycloalkyl group, or a heterocyclic group composed of 4 to 6 atoms, wherein C 4 -C 5 cycloalkyl group and the heterocyclic group composed of 4 to 6 atoms are each optionally substituted with 1-3 R 6 groups. In some embodiments, R 3 is H, -CH 3 , or -CF 3 ; and R 4 is H, -CH 3 , -CF 3 , cyclobutyl, or or R 3 and R 4 together with the carbon atom to which they are attached form wherein each group is each optionally substituted with 1-3 R 6 groups. In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached, and are substituted with 1 R 6 group that is methyl, to form In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached, and are substituted with 1 R 6 group, to form In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached, and are substituted with 1 R 6 group, to form In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached, and are substituted with 1 R 6 group, to form In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached, and are substituted with 2 R 6 groups, to form

[0039] In some embodiments, each R 6 is independently a C 1 -C 3 alkyl group, halogen, hydroxyl, -O(C1 -C 3 alkyl), -CN, C 1 -C 3 alkyl-CN, C 1 -C 3 alkyl-OH, or C 1 -C 3 haloalkyl; or two R groups attached to the same carbon atom together with the carbon atom to which they are attached form a spiro C 6 cycloalkyl, or a heterocyclic group consisting of 4 to 5 atoms in a spiro ring. In some embodiments, each R 3 -C 6 is independently -CH 6 , F, hydroxyl, -OCH 3 , -CN, -CH 3 , -CH 2 CN, -CH 2 OH, or -CF 3 ; or two R groups attached to the same carbon atom together with the carbon atom to which they are attached form a spirocyclopropyl. In some embodiments, R 6 and R 3 and R 4 together with the carbon atom to which they are attached form a cyclobutyl group which is substituted by two R 6 groups, and the two R 6 groups together with the carbon atom to which they are attached form a spirocyclopropyl, thereby forming

[0040] In some embodiments, R 5 is H, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, or C 3 -C 4 cycloalkyl. In some embodiments, R 5 is H, -CH 3 , -CHF 2 , or cyclopropyl.

[0041] In some embodiments, X is H, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, C 1 -C 3 alkyl-OH, C 1 -C 3 alkyl-CN, or C 8 optionally substituted by 1 - 3 R 3 -C 5Naphthenyl. In some embodiments, X is H or -CH 3 . In some embodiments, X is

[0042] In some embodiments, is a heterocyclic group composed of 4 to 6 atoms, or a heteroaryl group composed of 5 to 6 atoms, wherein the heterocyclic group or heteroaryl group each optionally contains 1 - 2 additional heteroatoms selected from the group consisting of N, O, and S, and wherein the heterocyclic group or heteroaryl group each optionally is substituted by 1 - 5 R 8 groups. In some embodiments, is a heterocyclic group composed of 4 to 5 atoms, or a heteroaryl group composed of 5 to 6 atoms, wherein the heterocyclic group or heteroaryl group each optionally contains 1 heteroatom selected from the group consisting of N and O, and wherein the heterocyclic group or heteroaryl group each optionally is substituted by 1 - 5 R 8 groups.

[0043] In some embodiments, X is

[0044] In some embodiments, Y is O. In some embodiments, Y is CH 2 , CHR 8 , or C(R 8 ) 2 .

[0045] In some embodiments, each R 7 is independently H, C 1 -C 3 alkyl, C 1 -C 3 alkyl-OH, or C 1 -C 3 haloalkyl; or two R 7 groups together with the carbon atom to which they are attached form C 3 -C 5 naphthenyl, or a heterocyclic group composed of 3 to 5 atoms. In some embodiments, each R 7 is independently H, -CH 3 , -CH 2 OH, or -CF 3 ; or two R 7 groups together with the carbon atom to which they are attached form cyclopropyl or oxetanyl.

[0046] In some embodiments, each R 8 is independently halogen, C 1 -C 3 alkyl, C 1 -C 3 alkyl-CN, C 1-C 3 alkyl-OH, C 1 -C 3 haloalkyl, -CN, oxo, or -O(C 1 -C 3 alkyl); or two R 8 groups together with the carbon atom to which they are attached form a spiro or fused C 3 -C 5 cycloalkyl, or a spiro or fused heterocyclic group consisting of 3 to 5 atoms. In some embodiments, each R 8 is independently F, -CH 3 -, -CH 2 CH 3 -, -CH 2 CN, -CH 2 OH, -CF 3 -, -CN, oxo, or -OCH 3 -; or two R 8 groups together with the carbon atom to which they are attached form a spiro or fused cyclopropyl, or a spiro or fused oxetanyl.

[0047] The present invention also discloses compounds selected from the compounds in Table 1, or their tautomers, or pharmaceutically acceptable salts of any of the above substances. The present invention also discloses compounds selected from any compounds disclosed above or disclosed in the present invention, or their tautomers, or pharmaceutically acceptable salts of any of the above substances, as well as pharmaceutically acceptable excipients.

[0048] The present invention also discloses a method for modulating the activity of immune cells, the method comprising contacting the immune cells with an effective amount of any compound disclosed above or disclosed in the present invention, or its tautomer, or a pharmaceutically acceptable salt of any of the above substances.

[0049] The present invention also discloses a method for treating cancer responsive to inhibition of Cbl-b activity in an individual in need thereof, the method comprising administering to the individual an effective amount of any compound disclosed above or disclosed in the present invention, or its tautomer, or a pharmaceutically acceptable salt of any of the above substances.

[0050] The present invention also discloses a method for inhibiting Cbl-b activity in an individual in need thereof, the method comprising administering to the individual an effective amount of any compound disclosed above or disclosed in the present invention, or its tautomer, or a pharmaceutically acceptable salt of any of the above substances.

[0051] The present invention also discloses a method for treating or preventing a disease or disorder associated with Cbl-b activity in an individual in need thereof, said method comprising administering to the individual any compound disclosed hereinabove or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt of any of the foregoing substances.

[0052] The present invention also discloses a method for generating modified immune cells, said method comprising culturing a cell population comprising immune cells in the presence of an effective amount of any compound disclosed hereinabove or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt of any of the foregoing substances.

[0053] The present invention also discloses modified immune cells comprising a Cbl-b inhibitor, wherein the Cbl-b inhibitor is any compound disclosed hereinabove or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt of any of the foregoing substances.

[0054] The present invention also discloses isolated modified immune cells, wherein the immune cells have been contacted with or in the presence of any compound disclosed hereinabove or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt of any of the foregoing substances.

[0055] The present invention also discloses a composition comprising a cell population, said cell population comprising isolated modified immune cells, wherein the immune cells have been contacted with or in the presence of any compound disclosed hereinabove or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt of any of the foregoing substances.

[0056] The present invention also discloses a method for inhibiting abnormal cell proliferation, said method comprising administering to an individual in need thereof an effective amount of isolated modified immune cells, wherein the immune cells have been contacted with or in the presence of any compound disclosed hereinabove or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt of any of the foregoing substances.

[0057] The present invention also discloses a method for inhibiting abnormal cell proliferation, said method comprising administering to an individual in need thereof a composition comprising a cell population containing isolated modified immune cells, wherein the immune cells have been contacted with or in the presence of any compound disclosed hereinabove or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt of any of the foregoing substances.

[0058] The present invention also discloses a method for inhibiting abnormal cell proliferation, said method comprising administering an effective amount of any compound disclosed hereinabove or disclosed in the present invention, or a tautomer thereof, or a pharmaceutically acceptable salt of any of the foregoing substances.

[0059] The present invention also discloses a cell culture composition, which comprises a cell population, the 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 of any of the foregoing substances.

[0060] The present invention also discloses a pharmaceutical composition, which comprises 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 of any of the foregoing substances.

[0061] The present invention also discloses an article, which comprises any modified immune cell disclosed in the present invention, any composition comprising the cell population disclosed in the present invention, any cell culture composition disclosed in the present invention, or any pharmaceutical composition disclosed in the present invention.

[0062] The present invention also discloses a kit, which comprises any modified immune cell disclosed in the present invention or any composition comprising the cell population disclosed in the present invention.

[0063] 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 disorder 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 of any of the foregoing substances.

[0064] In any embodiment disclosed in the present invention, the Cbl-b protein may be mammalian Cbl-b or human Cbl-b.

[0065] The present invention discloses a Cbl-b inhibitor compound, a vaccine and a composition comprising a Cbl-b inhibitor and a vaccine, as well as a method for using the same in treating cancer. The Cbl-b inhibitor and the cancer vaccine can be administered to an individual suffering from cancer.

[0066] In addition, the present invention discloses a Cbl-b inhibitor compound, an oncolytic virus and a composition comprising a Cbl-b inhibitor and an oncolytic virus, as well as a method for using the same in treating cancer. The Cbl-b inhibitor and the oncolytic virus can be administered to an individual suffering from cancer.

[0067] The present invention provides a method of immunization, which comprises administering an effective amount of a small molecule Cbl-b inhibitor to an individual in need thereof, and administering an effective amount of a vaccine to the individual.

[0068] The present invention provides a method for treating cancer, the method comprising administering to an individual suffering from cancer an effective amount of a small molecule Cbl-b inhibitor, and administering to the individual an effective amount of an oncolytic virus.

[0069] The present invention provides a method for treating cancer, the method comprising administering to an individual suffering from cancer an effective amount of an agent capable of reducing the activation threshold of immune cells, 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.

[0070] 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.

[0071] The present invention provides a kit for treating cancer, the kit comprising: (a) a small molecule Cbl-b inhibitor; (b) a therapeutic cancer vaccine; and (c) instructions for administering effective amounts of the Cbl-b inhibitor and the therapeutic cancer vaccine to treat cancer in an individual.

[0072] The present invention provides a kit for treating cancer, the kit 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.

[0073] 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.

[0074] The present invention provides a kit for treating cancer, the kit comprising: (a) a small molecule Cbl-b inhibitor; (b) an oncolytic virus; and (c) instructions for administering effective amounts of the Cbl-b inhibitor and the oncolytic virus to treat cancer in an individual.

[0075] The present invention provides a kit for treating cancer, the kit 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.

[0076] Furthermore, the present invention provides a method for treating a disease in a subject in need thereof by cell therapy, the method comprising administering to the individual an effective amount of one or more therapeutic cells to treat the disease; and administering to the subject an effective amount of a Cbl-b inhibitor, wherein the Cbl-b inhibitor is a compound described herein, and wherein the treatment by the therapeutic cells is enhanced by combination with the Cbl-b inhibitor.

[0077] BRIEF DESCRIPTION OF THE DRAWINGS

[0078] Figure 1 Shows the percentage of conditional survival of mice depicted by a Kaplan-Meier survival curve.

[0079] Figure 2 Shows the tumor volume of individual mice.

[0080] Figure 3 and Figure 4 Shows the tumor volume of an individual tumor. DETAILED DESCRIPTION OF THE INVENTION

[0082] The present invention provides compounds and pharmaceutical compositions that inhibit the Cbl-b enzyme, as well as methods of treatment using such compounds and pharmaceutical compositions. The compounds and compositions can be used for methods of modulating the immune system, treating diseases, and treating cells in vivo, in vitro, or ex vivo. In addition, the present invention provides pharmaceutical compositions comprising a cancer vaccine and a compound that inhibits the Cbl-b enzyme, as well as methods of treatment using such compounds, cancer vaccines, and pharmaceutical compositions. In addition, the present invention also provides pharmaceutical compositions comprising an oncolytic virus and a compound that inhibits the Cbl-b enzyme, as well as methods of treatment using such compounds, oncolytic viruses, and pharmaceutical compositions.

[0083] Both T cell activation and T cell tolerance are tightly controlled processes that regulate the immune response to tumors while preventing autoimmunity. Tolerance prevents the immune system from attacking cells expressing "self" or "autologous" antigens. During peripheral tolerance, T cells that recognize "self" or "autologous" antigens (i.e., autoreactive T cells) become functionally unresponsive or are deleted after encountering "self" or "autologous" antigens outside the thymus. Thus, the process of peripheral tolerance is important for preventing autoimmune diseases. Typically, 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 recognition and clearance by the immune system. The ability of cancer cells to avoid tumor immunosurveillance can lead to uncontrolled tumor growth. Thus, 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. Thus, treating T cell dysfunction, e.g., by increasing T cell activation, increasing T cell proliferation, reducing T cell tolerance, and / or reducing T cell exhaustion, may be beneficial for preventing or treating cancer. Other cells of the immune system are also important for recognizing and eliminating cancer cells during immunosurveillance. For example, natural killer (NK) cells are lymphocytes of the innate immune system that are capable of recognizing and killing cancer cells (see Martinez-Losato et al., Clin Cancer Res., 21:5048-5056, 2015). Recent studies have also shown that different subsets of B cells with distinct phenotypes and functions play different roles in the anti-tumor response (see Saravaria et al., Cell Mol Immunol., 14:662-674, 2017). Due to their roles in tumor surveillance, NK cells and B cells may also be suitable as therapeutic targets for preventing or treating cancer.

[0084] Cbl-b is a RING-type E3 ligase and plays an important role in the immune system due to its function as a negative regulator of immune activation. Cbl-b is important in reducing T cell activation and thereby enhancing or increasing T cell tolerance. Studies have found that Cbl-b-deficient T cells show a lower threshold for activation of antigen recognition receptors and co-stimulatory molecules (e.g., CD28). For example, the deletion of Cbl-b in T cells relieves the requirement 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, 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). To support this, the deletion of Cbl-b in cbl-b knockout mice results in impaired induction of T cell tolerance and exacerbation of autoimmunity (see Jeon et al., Immunity, 21:167-177, 2004). Importantly, the deletion of Cbl-b in mice also results in a strong anti-tumor response, which mainly depends on cytotoxic T cells. One study showed that cbl-b- / - CD8+ T cells are resistant to T regulatory cell-mediated inhibition 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 a mouse model (see Paolino et al., Nature, 507:508-512, 2014).

[0085] The present invention provides compounds and compositions that are both potent inhibitors of Cbl-b and can be used in new methods for treating diseases such as cancer. In some embodiments, the compounds and compositions provided by the present invention 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.

[0086] I. Definitions

[0087] The "effective amount" of the agent disclosed in the present invention is the amount sufficient to achieve a specific purpose. The "effective amount" can be determined in an empirical and conventional manner according to the said purpose. The "effective amount" or "sufficient amount" of the agent is the amount sufficient to produce the desired biological effect (such as a beneficial result, including a beneficial clinical result). In some embodiments, the term "effective amount" refers to the amount of the agent that effectively "treats" an individual (such as a mammal such as a human) of a disease or disorder.

[0088] The term "Cbl-b" as used in the present invention refers to the 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 (such as the ability to bind to E2 enzymes).

[0089] The terms "pharmaceutical formulation" and "pharmaceutical composition" refer to formulations in a form that enables the bioactivity of the active ingredient to function, and do 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. Such formulations or compositions may be sterile except for containing oncolytic viruses.

[0090] The "excipients" used in the present invention include pharmaceutically acceptable excipients, carriers, vehicles or stabilizers that are non-toxic to the cells or mammals exposed thereto at the doses and concentrations employed. Commonly physiologically acceptable excipients are pH-buffered aqueous solutions.

[0091] Referring to the compound described in the pharmaceutical composition, or claiming the compound described in the pharmaceutical composition claim, 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.

[0092] The term "treating" or "treatment" of a disease refers to implementing a protocol that may include administering to an individual (a human or other subject) one or more therapeutic agents in an effort to obtain a beneficial or desired result in the individual, including a clinical result. Beneficial or desired clinical results include, but are not limited to, alleviating or improving one or more symptoms, reducing the severity of the disease, stabilizing (i.e., not worsening) the disease state, preventing the spread of the disease, delaying or slowing the progression of the disease, improving or alleviating the disease state, and remission (partial or complete). "Treatment" can also refer to an extended survival period compared to the expected survival period of an untreated individual. In addition, "treating" and "treatment" can occur by administering a single dose of one or more therapeutic agents, or can occur when administering a series of doses of one or more therapeutic agents. "Treating" or "treatment" does not require complete alleviation of signs or symptoms, nor does it require a cure. "Treatment" can also refer to a clinical intervention, such as administering to an individual one or more therapeutic agents, aimed at altering the natural course of the individual or cell being treated (i.e., altering the course of the individual or cell that would occur in the absence of clinical intervention). The term "therapeutic agent" can refer to a Cbl-b inhibitor, a modified immune cell, or a composition thereof.

[0093] 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, sports, or pet animals such as dogs, horses, rabbits, cattle, pigs, hamsters, gerbils, mice, ferrets, rats, cats, and the like. In some embodiments, the individual or subject is a human.

[0094] 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 common elements of T cell exhaustion and / or T cell anergy, where antigen recognition may occur, but the subsequent immune response is ineffective in controlling tumor growth. The term "T cell dysfunction" also includes refractoriness or non-responsiveness to antigen recognition, e.g., impaired ability to translate antigen recognition into downstream T cell effector functions (such as proliferation, cytokine production, and / or target cell killing).

[0095] The term "T-cell anergy" refers to a state of non-responsiveness to antigenic stimulation due to incomplete or insufficient signaling through the T cell receptor. "T-cell anergy" can also be induced upon antigen stimulation in the absence of co-stimulation, rendering the cell refractory to subsequent activation by the antigen, even in the presence of co-stimulation.

[0096] The term "T cell exhaustion" refers to a state of T cell dysfunction caused by persistent TCR signaling that can occur during cancer. It differs from anergy in that it is not produced 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 distinct from functional effector T cells or memory T cells.

[0097] "T cell dysfunction" is a disease or disorder characterized by a reduced responsiveness of T cells to antigenic stimulation. Reduced responsiveness can 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 anergic or have a reduced ability to secrete cytokines, proliferate, or perform cytolytic activity.

[0098] "Enhancing / augmenting T cell function" refers to inducing, causing, or stimulating T cells to have a sustained or expanded biological function, or to reactivate or re-activate exhausted or inactivated T cells. Examples of enhanced or augmented T cell function include increased T cell activation (such as 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 state of T cells prior to treatment with a Cbl-b inhibitor. Methods for measuring enhanced or augmented T cell function are known in the art.

[0099] "Proliferation" as used in the present invention refers to the proliferation of cells. Increased proliferation includes producing a greater number of cells relative to a baseline value. Decreased proliferation includes producing a reduced 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.

[0100] "Alkyl" as used in the present invention refers to a saturated straight-chain (i.e., unbranched) or branched monovalent hydrocarbon chain or a combination thereof. Specific alkyl groups are those having a specified number of carbon atoms, e.g., having 1 to 20 carbon atoms ("C 1 -C 20 alkyl"), having 1 to 10 carbon atoms ("C 1 -C 10 " alkyl), having 1 to 8 carbon atoms ("C 1 -C 8 alkyl"), having 1 to 6 carbon atoms ("C 1 -C 6 alkyl"), having 2 to 6 carbon atoms ("C 2 -C 6 alkyl"), or having 1 to 4 carbon atoms ("C 1-C 4 The alkyl group of “alkyl”) The examples of the alkyl group include but are not limited to, homologue classes and isomer classes such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, and their similar groups.

[0101] The “alkenyl” used in the present invention refers to an unsaturated straight-chain (i.e., unbranched) or branched monovalent hydrocarbon chain or a combination thereof having at least one ethylenic unsaturated site (i.e., having at least one group moiety of the formula C═C). Specific alkenyl groups are those having a specified number of carbon atoms, for example, having 2 to 20 carbon atoms (“C 2 -C 20 alkenyl”), having 2 to 10 carbon atoms (“C 2 -C 10 ” alkenyl), having 2 to 8 carbon atoms (“C 2 -C 8 alkenyl”), having 2 to 6 carbon atoms (“C 2 -C 6 alkenyl”), or having 2 to 4 carbon atoms (“C 2 -C 4 alkenyl”) alkenyl groups. The alkenyl group can be in a “cis” or “trans” configuration, or in an “E” or “Z” configuration. Examples of alkenyl groups include but are not limited to groups such as vinyl (or ethenyl), 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, their homologue classes and isomer classes, etc.

[0102] The “alkynyl” used in the present invention refers to an unsaturated straight-chain (i.e., unbranched) or branched monovalent hydrocarbon chain or a combination thereof having at least one acetylenic unsaturated site (i.e., having at least one group moiety of the formula C≡C). Specific alkynyl groups are those having a specified number of carbon atoms, for example, having 2 to 20 carbon atoms (“C 2 -C 20 alkynyl”), having 2 to 10 carbon atoms (“C 2 -C 10 alkynyl”), having 2 to 8 carbon atoms (“C 2 -C 8 alkynyl”), having 2 to 6 carbon atoms (“C 2 -C 6 alkynyl”), or having 2 to 4 carbon atoms (“C 2 -C 4An alkynyl group of “alkynyl”) The examples of alkynyl groups include, but are not limited to, groups such as ethynyl (or ethinyl), prop-1-ynyl, prop-2-ynyl (or propargyl), but-1-ynyl, but-2-ynyl, but-3-ynyl, their homologs and isomers, etc.

[0103] “Alkylene” as used in the present invention refers to the same residue as an alkyl group but with a divalent nature. Specific alkylene groups are those having 1 to 6 carbon atoms (“C 1 -C 6 alkylene”), 1 to 5 carbon atoms (“C 1 -C 5 alkylene”), 1 to 4 carbon atoms (“C 1 -C 4 alkylene”), or 1 to 3 carbon atoms (“C 1 -C 3 alkylene”). Examples of alkylene groups include, but are not limited to, groups such as methylene (-CH 2 -), ethylene (-CH 2 CH 2 -), propylene (-CH 2 CH 2 CH 2 -), butylene (-CH 2 CH 2 CH 2 CH 2 -), etc.

[0104] “Cycloalkyl” as used in the present invention 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., cyclic) carbon atoms, for example, cycloalkyl groups having 3 to 12 ring carbon atoms (“C 3 -C 12 cycloalkyl”). Specific cycloalkyls are cyclic hydrocarbons having 3 to 8 ring carbon atoms (“C 3 -C 8 cycloalkyl”) or having 3 to 6 ring carbon atoms (“C 3 -C 6 cycloalkyl”). The cycloalkyl can be composed of one ring (such as cyclohexyl) or multiple rings (such as adamantyl), but does not include aryl (such as aromatic) groups. The cycloalkyl containing more than one ring can be fused, spiro, bridged, or a combination thereof. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl cyclobutyl cyclopentyl cyclohexyl 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl norbornanyl, and their similar groups.

[0105] The "subcycloalkyl group" used in the present invention refers to the same residue as the cycloalkyl group, but with a divalent nature. Specific subcycloalkyl groups are those having 3 to 12 ring carbon atoms ("C 3 -C 12 subcycloalkyl"), those having 3 to 8 ring carbon atoms ("C 3 -C 8 subcycloalkyl"), or those having 3 to 6 ring carbon atoms ("C 3 -C 6 subcycloalkyl"). Examples of subcycloalkyl groups include, but are not limited to, subcyclopropyl subcyclobutyl subcyclopentyl subcyclohexyl 1,2-subcyclohexenyl, 1,3-subcyclohexenyl, 1,4-subcyclohexenyl, subcycloheptyl subnorbornenyl, and similar groups.

[0106] The "aryl group" used in the present invention refers to an aromatic carbocyclic group having a single ring (e.g., phenyl) or multiple fused rings (e.g., naphthyl or anthracenyl), where one or more of the fused rings may not be aromatic. Specific aryl groups are those having 6 to 14 ring (i.e., cyclic) carbon atoms ("C 6 -C 14 aryl"). An aryl group having more than one ring (where at least one ring is non-aromatic) can be attached to the parent structure at an aromatic ring position or a non-aromatic ring position. In one variant, an aryl group having more than one ring (where at least one ring is non-aromatic) is attached to the parent structure at an aromatic ring position. Examples of aryl groups include, but are not limited to, groups such as phenyl, naphthyl, 1-naphthyl, 2-naphthyl, 1,2,3,4-tetrahydronaphthalen-6-yl and the like.

[0107] "Carbocyclic group" or "carbocycle" refers to an aromatic or non-aromatic monovalent cyclic group in which all ring members are carbon atoms, such as cyclohexyl, phenyl, 1,2-dihydronaphthyl, etc.

[0108] The "arylene group" used in the present invention refers to the same residue as the aryl group, but with a divalent nature. Specific arylene groups are those having 6 to 14 ring carbon atoms ("C 6 -C 14 arylene"). Examples of arylene groups include, but are not limited to, groups such as phenylene, orthophenylene (i.e., 1,2-phenylene), metaphenylene (i.e., 1,3-phenylene), paraphenylene (i.e., 1,4-phenylene), naphthylene, 1,2-naphthylene, 1,3-naphthylene, 1,4-naphthylene, 2,7-naphthylene, 2,6-naphthylene, and the like.

[0109] The "heteroaryl" used in the present invention refers to an unsaturated aromatic cyclic group having 1 to 14 ring carbon atoms and at least one ring heteroatom (including but not limited to heteroatoms such as nitrogen (N), oxygen (O), and sulfur (S)). The heteroaryl group can have a single ring (e.g., pyridyl or imidazolyl) or multiple fused rings (e.g., indolinyl, indolyl, or quinolinyl), where at least one fused ring is aromatic. Specific heteroaryl groups are rings composed 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 (N), oxygen (O), and sulfur (S) ("heteroaryl composed of 5 to 14 atoms"); rings composed 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 ("heteroaryl composed of 5 to 10 atoms"); or rings composed 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 ("heteroaryl composed of 5 to 7 atoms"). In one variant, the heteroaryl includes a monocyclic aromatic ring composed of 5, 6, or 7 atoms 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 variant, the heteroaryl includes a polycyclic aromatic ring having 1 to 12 ring carbon atoms and 1 to 6 ring heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. A heteroaryl group having more than one ring, where at least one ring is non-aromatic, can be attached to the parent structure at an aromatic ring position or a non-aromatic ring position. Examples of heteroaryl include but are not limited to groups such as pyridyl, benzimidazolyl, benzotriazolyl, benzothiophenyl, quinolinyl, indolyl, benzothiazolyl, etc. "Heteroaryl" also includes groups such as (2,4-dihydro-3H-1,2,4-triazol-3-one-2-yl), which has an aromatic tautomeric structure (1H-1,2,4-triazol-5-ol-1-yl).

[0110] As used herein, the terms "heterocyclic group" and "heterocyclic moiety" refer to non-aromatic, saturated or partially unsaturated cyclic groups having a specified number of ring atoms and heteroatoms, or, if no number of ring atoms or heteroatoms is specified, having 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. The heterocyclic moiety may 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, as well as bridged or spiro ring systems. Examples of heterocyclic moieties include but are not limited to aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, oxiranyl, oxetanyl, tetrahydrofuryl, tetrahydropyranyl, oxazolidinyl, piperazinyl, morpholinyl, dioxolanyl, 3,6-dihydro-2H-pyranyl, 2,3-dihydro-1H-imidazolyl, and the like.

[0111] As used herein, "heteroarylene" refers to the same residue as heteroaryl but having a divalent nature. Specific heteroarylene groups are rings composed of 5 to 14 atoms ("5- to 14-atom heteroarylene") having 1 to 12 ring carbon atoms and 1 to 6 ring heteroatoms independently selected from the group consisting of nitrogen, oxygen and sulfur; rings composed of 5 to 10 atoms ("5- to 10-atom heteroarylene") having 1 to 8 ring carbon atoms and 1 to 4 ring heteroatoms independently selected from the group consisting of nitrogen, oxygen and sulfur; or rings composed of 5, 6 or 7 atoms ("5- to 7-atom heteroarylene") having 1 to 5 ring carbon atoms and 1 to 4 ring heteroatoms independently selected from the group consisting of nitrogen, oxygen and sulfur. Examples of heteroarylene include but are not limited to groups such as pyridinylene, benzimidazolylene, benzotriazolylene, benzothiophenylene, quinolinylene, indolylene, benzothiazolylene and the like.

[0112] "Halogen" or "halo" refers to Group 17 elements having an atomic number from 9 to 85. Halo groups include fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

[0113] "Halogenated alkyl", "halogenated alkylene", "halogenated aryl", "halogenated arylene", "halogenated heteroaryl" and similar terms refer to a group moiety substituted by at least one halogen group. When a halogenated alkyl group moiety or other halogen-substituted group moiety is substituted by more than one halogen, it can be denoted by using a prefix corresponding to the number of the attached halogen group moieties. For example, dihaloaryl, dihaloalkyl, trihaloaryl, trihaloalkyl, etc., refer to aryl and alkyl groups substituted by 2 ("di") or 3 ("tri") halogen groups, which may or may not be the same halogen; thus, for example, the halogenated aryl group 4-chloro-3-fluorophenyl falls within the scope of dihaloaryl. A subset of the halogenated alkyl groups, in which each hydrogen (H) of the alkyl group is replaced by a halogen group, is called "perhalogenated alkyl". A specific perhalogenated alkyl group is trifluoromethyl (-CF 3 ). Similarly, "perhalogenated alkoxy" refers to an alkoxy group in which a halogen replaces each hydrogen (H) in the hydrocarbon of the alkyl group moiety constituting the alkoxy group. An example of perhalogenated alkoxy is trifluoromethoxy (-OCF 3 ). "Halogenated alkyl" includes mono-halogenated alkyl, dihalogenated alkyl, trihalogenated alkyl, perhalogenated alkyl, and any other possible number of halogenated substituents on the alkyl group; similarly, the same applies to other groups such as halogenated alkylene, halogenated aryl, halogenated arylene, halogenated heteroaryl, etc.

[0114] "Amino" refers to the group –NH 2 .

[0115] "Oxo" refers to the group =O, i.e., an oxygen atom double-bonded to carbon or other chemical elements.

[0116] Unless otherwise specified, "optionally substituted" means that the group is unsubstituted or substituted by one or more (e.g., 1, 2, 3, 4, or 5) substituents listed for the group, where 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 there are multiple substituents, unless otherwise specified, each substituent is independently selected. For example, each (C 1 -C 4 alkyl)(C 1 -C 4 alkyl) on the group -N(C 1 -C 4The (alkyl) substituents can each independently be selected such that groups such as –N(CH 3 )(CH 2 CH 3 ) are formed, and so on.

[0117] In addition to the disclosure of the present invention, when the term "substituted" is used to modify a particular group or moiety, it can also mean that one or more hydrogen atoms (H) of the particular group or moiety are each independently replaced by the same or different substituents as defined in the present invention. 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.

[0118] Unless otherwise specified, the substituents can be attached to any chemically possible position on the designated group or moiety. Thus, in one embodiment, -C 1 -C 8 alkyl-OH includes, for example, -CH 2 CH 2 OH, –CH(OH)-CH 3 , –CH 2 C(OH)(CH 3 ) 2 , and similar groups. As a further example, in one embodiment, -C 1 -C 6 alkyl-OH includes, for example, -CH 2 CH 2 OH, –CH(OH)-CH 3 , –CH 2 C(OH)(CH 3 ) 2 , and similar groups. As a further example, in one embodiment, -C 1 -C 6 alkyl-CN includes, for example, -CH 2 CH 2 CN, –CH(CN)-CH 3 , –CH 2 C(CN)(CH 3 ) 2 , and similar groups.

[0119] Unless a specific isotope of an element is specified in the general formula, the present disclosure includes all isotopomers of the compounds disclosed in the present invention, such as, for example, deuterated derivatives of the compounds (wherein H can be 2H, i.e., deuterium (D)). Deuterated compounds can provide favorable changes in pharmacokinetic (ADME) properties. Isotopologues can have isotope substitution at any or all positions in the structure, or can have atoms present at natural abundance at any or all positions in the structure.

[0120] The "small molecule" used in the present invention refers to a compound having a molecular weight of 1,000 daltons or less.

[0121] The hydrogen atom can also be replaced by a proximate bioisostere, such as fluorine, provided that such replacement results in a stable compound.

[0122] 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 the chemical structure or name, the structure or name is intended to encompass all possible stereoisomers of the described compound. Further, in the case of describing a particular stereochemical form, it is understood that all other stereochemical forms, as well as the general non-stereoselective forms and any mixtures in any ratio of the disclosed compounds, 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 included and described in the present invention. Compositions comprising the disclosed compounds are also contemplated, such as compositions of substantially pure compounds (including their specific stereochemical forms). Compositions of mixtures of the disclosed compounds in any ratio are also included in the present disclosure, including compositions of 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 included in the present invention. If stereochemistry is explicitly indicated for one or more portions of a molecule and not for another or other portions of the molecule, the structure is intended to cover all possible stereoisomers of the one or more portions for which stereochemistry is not explicitly indicated.

[0123] The present invention also includes any and all tautomeric forms of the compounds described herein.

[0124] 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 a medicine or drug 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) upon administration. Salts of the required basic compounds 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 compounds with amino acids, such as aspartates and glutamates, can also be prepared. Salts of the required acidic compounds 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, salts of alkali metals and alkaline earth metals, such as sodium salts, potassium salts, magnesium salts, 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 the acidic 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).

[0125] As described in Biological Examples 1, 8, and / or 12, for the determination of the IC 50 value of Cbl-b inhibition, the Cbl-b activity assay (Cbl-b inhibition assay) uses a mixture containing N-terminal biotinylated Avi-tagged Cbl-b (a fluorescently labeled inhibitor probe labeled with BODIPY FL (Example 54)) and an assay buffer. In one embodiment, the Cbl-b activity assay (Cbl-b inhibition assay) for determining the IC 50 of Cbl-b inhibition uses the conditions described in Biological Examples 1, 8, and / or 12, with 0.5 nM Cbl-b (“high” final concentration). In another embodiment, the Cbl-b activity assay (Cbl-b inhibition assay) for determining the IC 50The Cbl-b activity assay (Cbl-b inhibition assay) was performed using the conditions described in Biological Examples 1, 8, and / or 12, with 0.125 nM Cbl-b ("low" final concentration).

[0126] It should be understood that, for clarity, certain features of the present disclosure described in the context of separate embodiments 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, for the sake of brevity, also be provided separately or in any suitable sub-combination. All combinations of embodiments related to chemical groups represented by variables are specifically covered by and disclosed in the present invention as if each combination were separately and explicitly disclosed, insofar as such combinations include stable compounds (i.e., compounds that can be isolated, characterized, and tested for biological activity). In addition, all sub-combinations of chemical groups listed in the embodiments describing such variables are also specifically covered by and disclosed in the present invention as if each such sub-combination of chemical groups were separately and explicitly disclosed in the present invention.

[0127] It should be understood that aspects (cases) and embodiments described in the present invention as "comprising" or "including" or "covering" include embodiments of "consisting of" and "consisting essentially of".

[0128] The singular forms "a" and "the" as used in the present invention and the appended claims include the plural forms unless otherwise stated or the context clearly dictates otherwise. For example, "an" excipient or "the" excipient includes one or more excipients.

[0129] Reference to a value of "about" includes 90% to 110% of the stated value. For example, about 50 billion cells means 45 to 55 billion cells, including 50 billion cells. For example, a temperature of "about 100 degrees" means a temperature of about 90 degrees to about 110 degrees.

[0130] When a numerical range of compounds is given, all compounds within those numerical ranges are included, unless specifically excluded, including those designated as "a" and "b". For example, reference to compounds 9 - 13 means compound 9, compound 10, compound 11, compound 12, and compound 13.

[0131] II. Compounds

[0132] In one aspect, the present invention provides a compound represented by formula (I):

[0133]

[0134] or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0135]

[0136] Z 1 is CH or N;

[0137] Z 2 is CH or N;

[0138] R 1 is -CF 3 or cyclopropyl;

[0139] R 2 is -CF 3 or cyclopropyl;

[0140] R 3 is H, C 1 -C 2 alkyl, or C 1 -C 2 haloalkyl;

[0141] R 4 is H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, a heterocyclic group composed of 4 to 8 atoms, or C 3 -C 6 cycloalkyl, wherein the heterocyclic group or cycloalkyl group is each optionally substituted by 1 - 5 R 6 groups;

[0142] or R 3 and R 4 together with the carbon atom to which it is attached form a C 3 -C 5 cycloalkyl, or a heterocyclic group composed of 4 to 6 atoms, wherein C 3 -C 5 cycloalkyl and the heterocyclic group composed of 4 to 6 atoms are each optionally substituted by 1 - 5 R 6 groups;

[0143] R 5 is H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, or C 3 -C 6 cycloalkyl;

[0144] Each R 6 is independently C 1 -C 6 alkyl, halogen, hydroxy, -O(C 1 -C 6(alkyl), -CN, C 1 -C 6 alkyl-CN, C 1 -C 6 alkyl-OH, or C 1 -C 6 haloalkyl;

[0145] or two R groups attached to the same carbon atom together with the said carbon atom to which they are attached form a spiro C 6 -C 3 -C 6 cycloalkyl, or a heterocyclic group composed of 4 to 6 atoms;

[0146] X is H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkyl-OH, C 1 -C 6 alkyl-CN, optionally substituted by 1 - 5 R 8 groups of C 3 -C 6 cycloalkyl, or

[0147] is a heterocyclic group composed of 4 to 7 atoms, or a heteroaryl group composed of 5 to 8 atoms, wherein the heterocyclic group or heteroaryl group each optionally contains 1 - 2 additional heteroatoms selected from the group consisting of N, O, and S, and wherein the heterocyclic group or heteroaryl group each optionally is substituted by 1 - 5 R 8 groups;

[0148] Each R 7 is independently H, C 1 -C 6 alkyl, C 1 -C 6 alkyl-OH, or C 1 -C 6 haloalkyl;

[0149] or two R 7 groups together with the carbon atom to which they are attached form a C 3 -C 5 cycloalkyl, or a heterocyclic group composed of 3 to 5 atoms; and each R 8 is independently halogen, C 1 -C 6 alkyl, C 1 -C 6 alkyl-CN, C 1 -C 6Alkyl-OH, C 1 -C 6 haloalkyl, -CN, oxo, or -O(C 1 -C 6 alkyl);

[0150] Or two R 8 groups together with the carbon atom to which they are attached form a spiro or fused C 3 -C 5 cycloalkyl, or a spiro or fused heterocyclic group composed of 3 to 5 atoms.

[0151] In some embodiments, (i.e., the ring A group moiety) is In some embodiments, Z 1 is CH. In other embodiments, Z 1 is N. In some embodiments, R 1 is -CF 3 . In other embodiments, R 1 is cyclopropyl. 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 2 is -CF 3 . In other embodiments, R 2 is cyclopropyl. In some embodiments, the ring A group moiety is selected from the group consisting of:

[0152]

[0153] 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

[0154] In some embodiments, R 3 is H, C 1 -C 2 alkyl, or C 1 -C 2Halogenated alkyl. In some embodiments, R 3 is H, -CH 3 , or -CF 3 .

[0155] In some embodiments, R 3 is H.

[0156] In some embodiments, R 3 is C 1 -C 2 alkyl. In some embodiments, R 3 is methyl. In some embodiments, R 3 is ethyl.

[0157] In some embodiments, R 3 is C 1 -C 2 halogenated alkyl. In some embodiments, R 3 is C 1 -C 2 halogenated alkyl containing 1 - 5 halogen atoms. In some embodiments, R 3 is C 1 -C 2 halogenated alkyl containing 1 - 3 halogen atoms. In some embodiments, R 3 is C 1 halogenated alkyl. In some embodiments, R 3 is C 2 halogenated alkyl. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom, a bromine atom, and a fluorine atom. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom and a fluorine atom. In some embodiments, the halogen atoms are all fluorine atoms. In some embodiments, the halogen atoms are a combination of a chlorine atom and a fluorine atom. In some embodiments, R 3 is -CF 3 , -CCl 3 , -CF 2 Cl, -CFCl 2 , -CHF 2 , -CH 2 F, -CHCl 2 , -CH 2 Cl, or -CHFCl. In some embodiments, R 3 is -CF 3 .

[0158] In some embodiments, R 4 is H, C 1 -C 6 alkyl, C 1 -C 6A haloalkyl group, a heterocyclic group composed of 4 to 8 atoms, or a C 3 -C 6 cycloalkyl group, wherein the heterocyclic group or cycloalkyl group is each optionally substituted with 1-5 R 6 groups. In some embodiments, R 4 is H, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, a heterocyclic group composed of 4 to 6 atoms, or a C 4 -C 5 cycloalkyl group, wherein the heterocyclic group or cycloalkyl group is each optionally substituted with 1-3 R 6 groups. In some embodiments, R 4 is H, -CH 3 、-CF 3 、cyclobutyl, or

[0159] In some embodiments, R 4 is H.

[0160] In some embodiments, R 4 is C 1 -C 6 alkyl. In some embodiments, R 4 is C 1 -C 3 alkyl. In some embodiments, R 4 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 4 is -CH 3 .

[0161] In some embodiments, R 4 is C 1 -C 6 haloalkyl. In some embodiments, R 4 is C 1 -C 6 haloalkyl containing 1-7 halogen atoms. In some embodiments, R 4 is C 1 -C 3 haloalkyl. In some embodiments, R 4 is C 1 -C 3 haloalkyl containing 1-7 halogen atoms. In some embodiments, R 4 is C 1 -C 2Halogenated alkyl. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom, a bromine atom, and a fluorine atom. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom and a fluorine atom. In some embodiments, all of the halogen atoms are fluorine atoms. In some embodiments, the halogen atoms are a combination of a chlorine atom and a fluorine atom. In some embodiments, R 4 is -CF 3 -, -CCl 3 -, -CF 2 Cl, -CFCl 2 -, -CHF 2 -, -CH 2 F, -CHCl 2 -, -CH 2 F, or -CHFCl. In some embodiments, R 4 is -CF 3 .

[0162] In some embodiments, R 4 is a heterocyclic group consisting of 4 to 8 atoms optionally substituted with 1 to 5 R 6 groups. In some embodiments, R 4 is a heterocyclic group consisting of 4 to 6 atoms optionally substituted with 1 to 3 R 6 groups. In some embodiments, R 4 is a heterocyclic group consisting of 4 atoms optionally substituted with 1 to 2 R 6 groups. In some embodiments, the heterocyclic group is substituted with 5 R 6 groups. In some embodiments, the heterocyclic group is substituted with 4 R 6 groups. In some embodiments, the heterocyclic group is substituted with 3 R 6 groups. In some embodiments, the heterocyclic group is substituted with 2 R 6 groups. In some embodiments, the heterocyclic group is substituted with 1 R 6 group. In some embodiments, the heterocyclic group is unsubstituted. In some embodiments, the heterocyclic group contains 1 to 3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the heterocyclic group contains 1 nitrogen atom. In some embodiments, the heterocyclic group contains 2 nitrogen atoms. In some embodiments, the heterocyclic group contains 1 oxygen atom. In some embodiments, the heterocyclic group contains 2 oxygen atoms. In some embodiments, the heterocyclic group contains 1 oxygen atom and 1 nitrogen atom. In some embodiments, the heterocyclic group contains 1 sulfur atom. In some embodiments, the heterocyclic group contains 1 nitrogen atom and 1 sulfur atom. In some embodiments, R 4is oxetanyl, azetidinyl, tetrahydrofuranyl, dioxolanyl, pyrrolidinyl, pyrazolidinyl, piperidinyl, isoxazolidinyl, or tetrahydropyranyl, wherein each R 4 group is each optionally substituted with 1-5 R 6 groups. In some embodiments, R 4 is:

[0163]

[0164] In some embodiments, R 4 is

[0165] In some embodiments, R 4 is C 6 -C 3 cycloalkyl optionally substituted with 1-5 R 6 groups. In some embodiments, R 4 is C 6 -C 4 cycloalkyl optionally substituted with 1-3 R 5 groups. In some embodiments, the cycloalkyl is substituted with 5 R 6 groups. In some embodiments, the cycloalkyl is substituted with 4 R 6 groups. In some embodiments, the cycloalkyl is substituted with 3 R 6 groups. In some embodiments, the cycloalkyl is substituted with 2 R 6 groups. In some embodiments, the cycloalkyl is substituted with 1 R 6 group. In some embodiments, the cycloalkyl is unsubstituted. In some embodiments, R 4 is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, wherein each R 4 group is each optionally substituted with 1-5 R 6 groups. In some embodiments, R 4 is cyclopropyl or cyclobutyl. In some embodiments, R 4 is cyclobutyl.

[0166] In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached form C 3 -C 5 cycloalkyl or a heterocyclic group composed of 4 to 6 atoms, wherein C 3 -C 5 cycloalkyl and the heterocyclic group composed of 4 to 6 atoms are each optionally substituted with 1-5 R 6 groups. In some embodiments, R 3 and R4 together with the carbon atom to which it is attached forms a C 4 -C 5 cycloalkyl group, or a heterocyclic group composed of 4 to 6 atoms, where C 4 -C 5 the cycloalkyl group and the heterocyclic group composed of 4 to 6 atoms are each optionally substituted with 1 - 3 R 6 groups. In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached form where each group is each optionally substituted with 1 - 3 R 6 groups. In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached, and substituted with 1 R group that is a methyl group 6 form

[0167] In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached form a C 6 optionally substituted with 1 - 5 R 3 -C 5 cycloalkyl group. In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached form a C 6 optionally substituted with 1 - 3 R 4 -C 5 cycloalkyl group. In some embodiments, the cycloalkyl group is substituted with 5 R 6 groups. In some embodiments, the cycloalkyl group is substituted with 4 R 6 groups. In some embodiments, the cycloalkyl group is substituted with 3 R 6 groups. In some embodiments, the cycloalkyl group is substituted with 2 R 6 groups. In some embodiments, the cycloalkyl group is substituted with 1 R 6 group. In some embodiments, the cycloalkyl group is unsubstituted. In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached form where each group is each optionally substituted with 1 - 3 R 6 groups. In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached, and substituted with 1 R group that is a methyl group 6 form In some embodiments, The absolute stereochemistry at the carbon atom to which the methyl group is attached is (R)- (using the Cahn-Ingold-Prelog rules). In some embodiments, the absolute stereochemistry at the carbon atom to which the methyl group is attached is (S)-.

[0168] In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached form a heterocyclic group consisting of 4 to 6 atoms optionally substituted with 1 to 5 R 6 groups. In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached form a heterocyclic group consisting of 4 to 6 atoms optionally substituted with 1 to 3 R 6 groups. In some embodiments, the heterocyclic group is substituted with 5 R 6 groups. In some embodiments, the heterocyclic group is substituted with 4 R 6 groups. In some embodiments, the heterocyclic group is substituted with 3 R 6 groups. In some embodiments, the heterocyclic group is substituted with 2 R 6 groups. In some embodiments, the heterocyclic group is substituted with 1 R 6 group. In some embodiments, the heterocyclic group is unsubstituted. In some embodiments, the heterocyclic group contains 1 to 3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the heterocyclic group contains 1 nitrogen atom. In some embodiments, the heterocyclic group contains 2 nitrogen atoms. In some embodiments, the heterocyclic group contains 1 oxygen atom. In some embodiments, the heterocyclic group contains 2 oxygen atoms. In some embodiments, the heterocyclic group contains 1 oxygen atom and 1 nitrogen atom. In some embodiments, the heterocyclic group contains 1 sulfur atom. In some embodiments, the heterocyclic group contains 1 nitrogen atom and 1 sulfur atom. In some embodiments, R 4 is oxetanyl, azetidinyl, tetrahydrofuranyl, dioxolanyl, pyrrolidinyl, pyrazolidinyl, piperidinyl, isoxazolidinyl, or tetrahydropyranyl, wherein each group of R 4 is optionally substituted with 1 to 5 R 6 groups. In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached form wherein each group is optionally substituted with 1 to 3 R 6 groups. In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached form In some embodiments, R 3 and R 4 together with the carbon atom to which they are attached form

[0169] In some embodiments, each R 6 is independently C 1 -C 6 alkyl, halogen, hydroxyl, -O(C 1 -C 6 alkyl), -CN, C 1 -C 6 alkyl-CN, C 1 -C 6 alkyl-OH, or C 1 -C 6 haloalkyl. In some embodiments, each R 6 is independently C 1 -C 3 alkyl, halogen, hydroxyl, -O(C 1 -C 3 alkyl), -CN, C 1 -C 3 alkyl-CN, C 1 -C 3 alkyl-OH, or C 1 -C 3 haloalkyl. In some embodiments, each R 6 is independently -CH 3 、fluorine (F), hydroxyl, -OCH 3 、-CN, -CH 2 CN, -CH 2 OH, or -CF 3 .

[0170] In some embodiments, R 6 is C 1 -C 6 alkyl. In some embodiments, R 6 is C 1 -C 3 alkyl. In some embodiments, R 6 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 6 is -CH 3 .

[0171] In some embodiments, R 6 is halogen. In some embodiments, R 6 is chlorine, fluorine, or bromine. In some embodiments, R 6 is chlorine or fluorine. In some embodiments, R 7 is fluorine.

[0172] In some embodiments, R 6 is a hydroxyl group.

[0173] In some embodiments, R 6 is -O(C 1 -C 6 alkyl). In some embodiments, R 6 is -O-(C 1 -C 3 alkyl). In some embodiments, R 6 is -O(methyl), -O(ethyl), -O(n-propyl), or -O(isopropyl). In some embodiments, R 6 is -OCH 3 or -OCH 2 CH 3 ). In some embodiments, R 6 is -OCH 3 .

[0174] In some embodiments, R 6 is -CN. In some embodiments, R 6 is C 1 -C 6 alkyl-CN. In some embodiments, R 6 is C 1 -C 3 alkyl-CN. In some embodiments, R 6 is -CH 2 CN, -CH 2 CH 2 -CN, -CH 2 CH 2 CH 2 -CN, or -C(CH 3 ) 2 -CN. In some embodiments, R 6 is -CH 2 CN.

[0175] In some embodiments, R 6 is C 1 -C 6 alkyl-OH. In some embodiments, R 6 is C 1 -C 3 alkyl-OH. In some embodiments, R 6 is -CH 2 OH, -CH 2 CH 2 -OH, -CH 2 CH 2 CH2 -OH, or -C(CH 3 ) 2 -OH. In some embodiments, R 6 is -CH 2 OH.

[0176] In some embodiments, R 6 is C 1 -C 6 haloalkyl. In some embodiments, R 6 is C 1 -C 6 haloalkyl containing 1 to 7 halogen atoms. In some embodiments, R 6 is C 1 -C 3 haloalkyl. In some embodiments, R 6 is C 1 -C 3 haloalkyl containing 1 to 7 halogen atoms. In some embodiments, R 6 is C 1 -C 3 haloalkyl containing 1 to 5 halogen atoms. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom, a bromine atom, and a fluorine atom. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom and a fluorine atom. In some embodiments, the halogen atoms are all fluorine atoms. In some embodiments, the halogen atoms are a combination of a chlorine atom and a fluorine atom. In some embodiments, R 6 is -CF 3 , -CCl 3 , -CF 2 Cl, -CFCl 2 , -CHF 2 , -CH 2 F, -CHCl 2 , -CH 2 Cl, or -CHFCl. In some embodiments, R 6 is -CF 3 .

[0177] In some embodiments, two R 6 groups attached to the same carbon atom together with the carbon atom to which they are attached form a spiro C 3 -C 6 cycloalkyl, or a heterocyclic group consisting of 4 to 6 atoms in a spiro ring. In some embodiments, two R 6 groups attached to the same carbon atom together with the carbon atom to which they are attached form a spiro C 3 -C 6 cycloalkyl, or a heterocyclic group consisting of 4 to 5 atoms in a spiro ring.

[0178] In some embodiments, two R 6 The group together with the carbon atom to which it is attached forms a spiro 3 -C 6 In some embodiments, two R 6 The group together with the carbon atom to which it is attached forms a spiro 3 -C 5 In some embodiments, two R 6 The group together with the carbon atom to which it is attached forms a spiro 3 -C 4 In some embodiments, two R 6 The group together with the carbon atom to which it is attached forms a spirocyclopropyl, spirocyclobutyl, or spirocyclopentyl. In some embodiments, two R 6 The group together with the carbon atom to which it is attached forms a spirocyclopropyl group.

[0179] In some embodiments, two R 6 The group together with the carbon atom to which it is attached forms a spiro heterocyclic group consisting of 4 to 6 atoms. In some embodiments, two R attached to the same carbon atom 6 The group together with the carbon atom to which it is attached forms a spiro heterocyclic group consisting of 4 to 5 atoms. In some embodiments, the heterocyclic group contains 1-3 heteroatoms selected from the group consisting of N, O and S. In some embodiments, the heterocyclic group contains 1 nitrogen atom. In some embodiments, the heterocyclic group contains 2 nitrogen atoms. In some embodiments, the heterocyclic group contains 1 oxygen atom. In some embodiments, the heterocyclic group contains 2 oxygen atoms. In some embodiments, the heterocyclic group contains 1 oxygen atom and 1 nitrogen atom. In some embodiments, the heterocyclic group contains 1 sulfur atom. In some embodiments, the two R attached to the same carbon atom 6 The radical, together with the carbon atom to which it is attached, forms a spirooxetanyl, spiroazetidinyl, spirotetrahydrofuranyl, spirodioxolanyl, spiropyrrolidinyl, spiropyrazolidinyl, spiropiperidinyl, spiroisoxazolidinyl, or spirotetrahydropyranyl radical.

[0180] In some embodiments, R 5 It is H, C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, or C 3-C 6 Cycloalkyl. In some embodiments, R 5 is H, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, or C 3 -C 4 cycloalkyl. In some embodiments, R 5 is H, -CH 3 , -CHF 2 , or cyclopropyl.

[0181] In some embodiments, R 5 is H.

[0182] In some embodiments, R 5 is C 1 -C 6 alkyl. In some embodiments, R 5 is C 1 -C 3 alkyl. In some embodiments, R 5 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 5 is -CH 3 .

[0183] In some embodiments, R 5 is C 1 -C 6 haloalkyl. In some embodiments, R 5 is C 1 -C 6 haloalkyl containing 1-7 halogen atoms. In some embodiments, R 5 is C 1 -C 3 haloalkyl. In some embodiments, R 5 is C 1 -C 3 haloalkyl containing 1-7 halogen atoms. In some embodiments, R 5 is C 1 -C 3 haloalkyl containing 1-5 halogen atoms. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom, a bromine atom, and a fluorine atom. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom and a fluorine atom. In some embodiments, the halogen atoms are all fluorine atoms. In some embodiments, the halogen atoms are a combination of a chlorine atom and a fluorine atom. In some embodiments, R 5 is -CF 3 , -CCl 3 , -CF2 Cl, -CFCl 2 , -CHF 2 , -CH 2 F, -CHCl 2 , -CH 2 Cl, or -CHFCl. In some embodiments, R 5 is -CHF 2 .

[0184] In some embodiments, R 5 is C 3 -C 6 cycloalkyl. In some embodiments, R 5 is C 3 -C 5 cycloalkyl. In some embodiments, R 5 is C 3 -C 4 cycloalkyl. In some embodiments, R 5 is cyclopropyl, cyclobutyl, or cyclopentyl. In some embodiments, R 5 is cyclopropyl.

[0185] In some embodiments, X is H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkyl-OH, C 1 -C 6 alkyl-CN, C 8 optionally substituted with 1-5 R 3 -C 6 cycloalkyl groups. In some embodiments, X is H, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, C 1 -C 3 alkyl-OH, C 1 -C 3 alkyl-CN, C 8 optionally substituted with 1-3 R 3 -C 5 cycloalkyl groups. In some embodiments, X is H, or -CH 3 .

[0186] In some embodiments, X is H.

[0187] In some embodiments, X is C 1 -C 6Alkyl. In some embodiments, X is C 1 -C 3 Alkyl. In some embodiments, X is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, X is -CH 3 .

[0188] In some embodiments, X is C 1 -C 6 Haloalkyl. In some embodiments, X is C 1 -C 6 Haloalkyl. In some embodiments, X is C 1 -C 3 Haloalkyl. In some embodiments, X is C 1 -C 3 Haloalkyl. In some embodiments, X is C 1 -C 3 Haloalkyl. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom, a bromine atom, and a fluorine atom. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom and a fluorine atom. In some embodiments, the halogen atoms are all fluorine atoms. In some embodiments, the halogen atoms are a combination of a chlorine atom and a fluorine atom. In some embodiments, X is -CF 3 , -CCl 3 , -CF 2 Cl, -CFCl 2 , -CHF 2 , -CH 2 F, -CHCl 2 , -CH 2 Cl, or -CHFCl. In some embodiments, X is -CF 3 .

[0189] In some embodiments, X is C 1 -C 6 Alkyl-OH. In some embodiments, X is C 1 -C 3 Alkyl-OH. In some embodiments, X is -CH 2 OH, -CH 2 CH 2 -OH, -CH 2 CH 2 CH 2 -OH, or -C(CH 3 ) 2 -OH. In some embodiments, X is -CH 2 OH.

[0190] In some embodiments, X is C 1 -C 6 alkyl-CN. In some embodiments, X is C 1 -C 3 alkyl-CN. In some embodiments, X is -CH 2 CN, -CH 2 CH 2 -CN, -CH 2 CH 2 CH 2 -CN, or -C(CH 3 ) 2 -CN. In some embodiments, X is -CH 2 CN.

[0191] In some embodiments, X is C optionally substituted with 1-5 R 8 groups and is 3 -C 6 cycloalkyl. In some embodiments, X is C optionally substituted with 1-3 R 8 groups and is 3 -C 5 cycloalkyl. In some embodiments, the cycloalkyl is substituted with 5 R 8 groups. In some embodiments, the cycloalkyl is substituted with 4 R 8 groups. In some embodiments, the cycloalkyl is substituted with 3 R 8 groups. In some embodiments, the cycloalkyl is substituted with 2 R 8 groups. In some embodiments, the cycloalkyl is substituted with 1 R 8 group. In some embodiments, the cycloalkyl is unsubstituted. In some embodiments, X is cyclopropyl, cyclobutyl, or cyclopentyl, where each group of X is optionally substituted with 1-5 R 8 groups. In some embodiments, X is cyclopropyl.

[0192] In some embodiments, X is where the ring B group moiety, as shown, is a heterocyclic group composed of 4 to 7 atoms or a heteroaryl group composed of 5 to 8 atoms, where the heterocyclic group or heteroaryl group each optionally contains 1-2 additional heteroatoms selected from the group consisting of N, O, and S, and where the heterocyclic group or heteroaryl group each is optionally substituted with 1-5 R 8Group substitution. In some embodiments, the ring B group moiety is a heterocyclic group composed of 4 to 6 atoms, or a heteroaryl group composed of 5 to 6 atoms, wherein the heterocyclic group or heteroaryl group each optionally contains 1-2 additional heteroatoms selected from the group consisting of N, O, and S, and wherein the heterocyclic group or heteroaryl group each is optionally substituted with 1-5 R 8 Group substitution. In some embodiments, the ring B group moiety is a heterocyclic group composed of 4 to 5 atoms, or a heteroaryl group composed of 5 to 6 atoms, wherein the heterocyclic group or heteroaryl group each optionally contains 1 additional heteroatom selected from the group consisting of N and O, and wherein the heterocyclic group or heteroaryl group each is optionally substituted with 1-5 R 8 Group substitution.

[0193] In some embodiments, the ring B group moiety is a heterocyclic group composed of 4 to 7 atoms, the heterocyclic group composed of 4 to 7 atoms optionally contains 1-2 additional heteroatoms selected from the group consisting of N, O, and S, wherein the heterocyclic group is optionally substituted with 1-5 R 8 Group substitution. In some embodiments, the ring B group moiety is a heterocyclic group composed of 4 to 6 atoms, the heterocyclic group composed of 4 to 6 atoms optionally contains 1-2 additional heteroatoms selected from the group consisting of N, O, and S, wherein the heterocyclic group is optionally substituted with 1-5 R 8 Group substitution. In some embodiments, the ring B group moiety is a heterocyclic group composed of 4 to 5 atoms, the heterocyclic group composed of 4 to 5 atoms optionally contains 1 additional heteroatom selected from the group consisting of N and O, wherein the heterocyclic group is optionally substituted with 1-5 R 8 Group substitution. In some embodiments, the heterocyclic group is substituted with 5 R 8 Group substitution. In some embodiments, the heterocyclic group is substituted with 4 R 8 Group substitution. In some embodiments, the heterocyclic group is substituted with 3 R 8 Group substitution. In some embodiments, the heterocyclic group is substituted with 2 R 8 Group substitution. In some embodiments, the heterocyclic group is substituted with 1 R 8Group substitution. In some embodiments, the heterocyclic group is unsubstituted. In some embodiments, the heterocyclic group contains 1-2 additional heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the heterocyclic group contains 1 additional nitrogen atom. In some embodiments, the heterocyclic group contains 2 additional nitrogen atoms. In some embodiments, the heterocyclic group further contains 1 oxygen atom. In some embodiments, the heterocyclic group further contains 2 oxygen atoms. In some embodiments, the heterocyclic group further contains 1 oxygen atom and 1 nitrogen atom. In some embodiments, the heterocyclic group further contains 1 sulfur atom. In some embodiments, the heterocyclic group contains no additional heteroatoms. In some embodiments, the heterocyclic group is azetidinyl, pyrrolidinyl, pyrazolidinyl, piperidinyl, or isoxazolidinyl, wherein each group of the heterocyclic group is optionally substituted by 1-5 R 8 groups.

[0194] In some embodiments, the ring B group moiety is a heteroaryl composed of 5 to 8 atoms, and the heteroaryl composed of 5 to 8 atoms optionally contains 1-2 additional heteroatoms selected from the group consisting of N, O, and S, wherein the heteroaryl is optionally substituted by 1-5 R 8 groups. In some embodiments, the ring B group moiety is a heteroaryl composed of 5 to 6 atoms, and the heteroaryl composed of 5 to 6 atoms optionally contains 1-2 additional heteroatoms selected from the group consisting of N, O, and S, wherein the heteroaryl is optionally substituted by 1-5 R 8 groups. In some embodiments, the ring B group moiety is a heteroaryl composed of 5 to 6 atoms, and the heteroaryl composed of 5 to 6 atoms optionally contains 1 additional heteroatom selected from the group consisting of N and O, wherein the heteroaryl is optionally substituted by 1-5 R 8 groups. In some embodiments, the heteroaryl is substituted by 5 R 8 groups. In some embodiments, the heteroaryl is substituted by 4 R 8 groups. In some embodiments, the heteroaryl is substituted by 3 R 8 groups. In some embodiments, the heteroaryl is substituted by 2 R 8 groups. In some embodiments, the heteroaryl is substituted by 1 R 8Group substitution. In some embodiments, the heteroaryl is unsubstituted. In some embodiments, the heteroaryl contains 1-2 additional heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the heteroaryl contains 1 additional nitrogen atom. In some embodiments, the heteroaryl contains 2 additional nitrogen atoms. In some embodiments, the heteroaryl further contains 1 oxygen atom. In some embodiments, the heteroaryl further contains 2 oxygen atoms. In some embodiments, the heteroaryl further contains 1 oxygen atom and 1 additional nitrogen atom. In some embodiments, the heteroaryl further contains 1 sulfur atom. In some embodiments, the heteroaryl contains no additional heteroatoms. In some embodiments, the heteroaryl is pyrrolyl, imidazolyl, pyrazolyl, isoxazolyl, oxazolyl, thiazolyl, isothiazolyl, triazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, or triazolyl, wherein each group of the heteroaryl is optionally substituted by 1-5 R 8 groups.

[0195] In some embodiments, the ring B group moiety is wherein Y is O, CH 2 , CHR 8 , or C(R 8 ) 2 , and X is In some embodiments, Y is oxygen (O). In other embodiments, Y is CH 2 , CHR 8 , or C(R 8 ) 2 . In some embodiments, Y is CH 2 . In some embodiments, Y is CHR 8 . In some embodiments, Y is C(R 8 ) 2 . In some embodiments, the ring B group moiety is substituted by a total of 1-5 R 8 groups. In some embodiments, the ring B group moiety is substituted by a total of 1-3 R 8 groups. Thus, if Y is CHR 8 , then the ring B group moiety can be substituted by up to 4 additional R 8 groups. Similarly, if Y is CH(R 8 ) 2 , then the ring B group moiety can be substituted by up to 3 additional R 8 groups. In some embodiments, the ring B group moiety is substituted by 5 R 8 groups. In some embodiments, the ring B group moiety is substituted by 4 R 8 groups. In some embodiments, the ring B group moiety is substituted by 3 R 8Group substitution. In some embodiments, the ring B group moiety is substituted with two R 8 groups. In some embodiments, the ring B group moiety is substituted with one R 8 group. In some embodiments, the ring B group moiety is unsubstituted. In some embodiments, the ring B group moiety is where each R 8 is independently as described in the present invention.

[0196] In some embodiments, each R 7 is independently H, C 1 -C 6 alkyl, C 1 -C 6 alkyl-OH, or C 1 -C 6 haloalkyl. In some embodiments, each R 7 is independently H, C 1 -C 3 alkyl, C 1 -C 3 alkyl-OH, or C 1 -C 3 haloalkyl. In some embodiments, each R 7 is independently H, -CH 3 、-CH 2 OH, or -CF 3 .

[0197] In some embodiments, both R 7 groups are hydrogen (H). In some embodiments, one R 7 group is H. In some embodiments, one R 7 group is H and the other R 7 group is C 1 -C 6 alkyl, C 1 -C 6 alkyl-OH, or C 1 -C 6 haloalkyl. In some embodiments, one R 7 group is H and the other R 7 group is C 1 -C 6 alkyl. In some embodiments, one R 7 group is H and the other R 7 group is C 1 -C 3 alkyl. In some embodiments, one R 7 group is H and the other R 7 group is -CH3 .

[0198] In some embodiments, R 7 is C 1 -C 6 alkyl. In some embodiments, R 7 is C 1 -C 3 alkyl. In some embodiments, R 7 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, one R 7 group is methyl, ethyl, n-propyl, or isopropyl, and the other R 7 group is H. In some embodiments, R 7 is -CH 3 .

[0199] In some embodiments, R 7 is C 1 -C 6 alkyl-OH. In some embodiments, R 7 is C 1 -C 3 alkyl-OH. In some embodiments, R 7 is -CH 2 OH, -CH 2 CH 2 -OH, -CH 2 CH 2 CH 2 -OH, or -C(CH 3 ) 2 -OH. In some embodiments, R 7 is -CH 2 OH. In some embodiments, one R 7 group is C 1 -C 6 alkyl-OH, and the other R 7 group is H. In some embodiments, one R 7 group is C 1 -C 3 alkyl-OH, and the other R 7 group is H. In some embodiments, one R 7 group is -CH 2 OH, -CH 2 CH 2 -OH, -CH 2 CH 2 CH 2 -OH, or -C(CH 3 ) 2 -OH, and the other R 7The group is H. In some embodiments, one R 7 group is -CH 2 OH, and the other R 7 group is H.

[0200] In some embodiments, R 7 is C 1 -C 6 haloalkyl. In some embodiments, R 7 is C 1 -C 6 haloalkyl containing 1 to 7 halogen atoms. In some embodiments, R 7 is C 1 -C 3 haloalkyl. In some embodiments, R 7 is C 1 -C 3 haloalkyl containing 1 to 7 halogen atoms. In some embodiments, R 7 is C 1 -C 3 haloalkyl containing 1 to 5 halogen atoms. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom, a bromine atom, and a fluorine atom. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom and a fluorine atom. In some embodiments, the halogen atoms are all fluorine atoms. In some embodiments, the halogen atoms are a combination of a chlorine atom and a fluorine atom. In some embodiments, R 7 is -CF 3 、-CCl 3 、-CF 2 Cl、-CFCl 2 、-CHF 2 、-CH 2 F、-CHCl 2 、-CH 2 F、or -CHFCl. In some embodiments, R 7 is -CF 3 。

[0201] In some embodiments, the two R 7 groups together with the carbon atom to which they are attached form a C 3 -C 5 cycloalkyl, or a heterocyclic group consisting of 3 to 5 atoms. In some embodiments, the two R 7 groups together with the carbon atom to which they are attached form a cyclopropyl or an oxetanyl group.

[0202] In some embodiments, the two R 7 groups together with the carbon atom to which they are attached form a C 3 -C5 Cycloalkyl. In some embodiments, two R 7 groups together with the carbon atom to which they are attached form cyclopropyl or cyclobutyl. In some embodiments, two R 7 groups together with the carbon atom to which they are attached form cyclopropyl.

[0203] In some embodiments, two R 7 groups together with the carbon atom to which they are attached form a heterocyclic group composed of 3 to 5 atoms. In some embodiments, two R 7 groups together with the carbon atom to which they are attached form a heterocyclic group composed of 3 to 4 atoms. In some embodiments, the heterocyclic group contains 1 - 3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the heterocyclic group contains 1 nitrogen atom. In some embodiments, the heterocyclic group contains 2 nitrogen atoms. In some embodiments, the heterocyclic group contains 1 oxygen atom. In some embodiments, the heterocyclic group contains 2 oxygen atoms. In some embodiments, the heterocyclic group contains 1 oxygen atom and 1 nitrogen atom. In some embodiments, the heterocyclic group contains 1 sulfur atom. In some embodiments, the heterocyclic group contains 1 nitrogen atom and 1 sulfur atom. In some embodiments, R 7 is aziridinyl, oxiranyl, oxetanyl, azetidinyl, tetrahydrofuryl, dioxolanyl, pyrrolidinyl, pyrazolidinyl, or isoxazolidinyl.

[0204] In some embodiments, each R 8 is independently halogen, C 1 -C 6 alkyl, C 1 -C 6 alkyl-CN, C 1 -C 6 alkyl-OH, C 1 -C 6 haloalkyl, -CN, oxo, or -O(C 1 -C 6 alkyl). In some embodiments, each R 8 is independently halogen, C 1 -C 3 alkyl, C 1 -C 3 alkyl-CN, C 1 -C 3 alkyl-OH, C 1 -C 3 haloalkyl, -CN, oxo, or -O(C 1 -C 3 alkyl). In some embodiments, each R 8Each independently is fluorine (F), -CH 3 , -CH 2 CH 3 , -CH 2 CN, -CH 2 OH, -CF 3 , -CN, oxo, or -OCH 3 .

[0205] In some embodiments, R 8 is a halogen. In some embodiments, R 8 is chlorine, fluorine, or bromine. In some embodiments, R 8 is chlorine or fluorine. In some embodiments, R 8 is fluorine.

[0206] In some embodiments, R 8 is C 1 -C 6 alkyl. In some embodiments, R 8 is C 1 -C 3 alkyl. In some embodiments, R 8 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 8 is -CH 3 or -CH 2 CH 3 .

[0207] In some embodiments, R 8 is -CN. In some embodiments, R 8 is C 1 -C 6 alkyl-CN. In some embodiments, R 8 is C 1 -C 3 alkyl-CN. In some embodiments, R 8 is -CH 2 CN, -CH 2 CH 2 -CN, -CH 2 CH 2 CH 2 -CN, or -C(CH 3 ) 2 -CN. In some embodiments, R 8 is -CH 2 CN.

[0208] In some embodiments, R 8 is C 1 -C 6 alkyl-OH. In some embodiments, R 8is C 1 -C 3 alkyl-OH. In some embodiments, R 8 is -CH 2 OH, -CH 2 CH 2 -OH, -CH 2 CH 2 CH 2 -OH, or -C(CH 3 ) 2 -OH. In some embodiments, R 8 is -CH 2 OH.

[0209] In some embodiments, R 8 is C 1 -C 6 haloalkyl. In some embodiments, R 8 is C 1 -C 6 haloalkyl containing 1-7 halogen atoms. In some embodiments, R 8 is C 1 -C 3 haloalkyl. In some embodiments, R 8 is C 1 -C 3 haloalkyl containing 1-7 halogen atoms. In some embodiments, R 8 is C 1 -C 3 haloalkyl containing 1-5 halogen atoms. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom, a bromine atom, and a fluorine atom. In some embodiments, the halogen atoms are each independently selected from the group consisting of a chlorine atom and a fluorine atom. In some embodiments, the halogen atoms are all fluorine atoms. In some embodiments, the halogen atoms are a combination of a chlorine atom and a fluorine atom. In some embodiments, R 8 is -CF 3 , -CCl 3 , -CF 2 Cl, -CFCl 2 , -CHF 2 , -CH 2 F, -CHCl 2 , -CH 2 F, or -CHFCl. In some embodiments, R 8 is -CF 3 .

[0210] In some embodiments, R 8 is oxo.

[0211] In some embodiments, R 8 is -O(C 1 -C 6 alkyl). In some embodiments, R 8 is -O(C 1 -C 3 alkyl). In some embodiments, R 8 is -O(methyl), -O(ethyl), -O(n-propyl), or -O(isopropyl). In some embodiments, R 8 is -OCH 3 or -OCH 2 CH 3 ). In some embodiments, R 8 is -OCH 3 .

[0212] In some embodiments, the two R 8 groups together with the carbon atom to which they are attached form a spiro or fused C 3 -C 5 cycloalkyl, or a spiro or fused heterocyclic group consisting of 3 to 5 atoms. In some embodiments, the two R 8 groups together with the carbon atom to which they are attached form a spiro or fused cyclopropyl, or a spiro or fused oxetanyl.

[0213] In some embodiments, the two R 8 groups together with the carbon atom to which they are attached form a spiro or fused C 3 -C 5 cycloalkyl. In some embodiments, the two R 8 groups together with the carbon atom to which they are attached form a spiro C 3 -C 5 cycloalkyl. In some embodiments, the two R 8 groups together with the carbon atom to which they are attached form a spiro cyclopropyl. In some embodiments, the two R 8 groups together with the carbon atom to which they are attached form a spiro cyclobutyl. In some embodiments, the two R 8 groups together with the carbon atom to which they are attached form a spiro cyclopentyl. In some embodiments, the two R 8 groups together with the carbon atom to which they are attached form a fused C 3 -C 5 cycloalkyl. In some embodiments, the two R 8 groups together with the carbon atom to which they are attached form a fused cyclopropyl. In some embodiments, the two R 8 groups together with the carbon atom to which they are attached form a fused cyclobutyl. In some embodiments, the two R 8The group together with the carbon atom to which it is attached forms a fused cyclopentyl group. In some embodiments, two R 8 groups together with the carbon atom to which they are attached form a spiro or fused cyclopropyl group.

[0214] In some embodiments, two R 8 groups together with the carbon atom to which they are attached form a spiro or fused heterocyclic group composed of 3 to 5 atoms. In some embodiments, two R 8 groups together with the carbon atom to which they are attached form a spiro heterocyclic group composed of 3 to 5 atoms. In some embodiments, two R 8 groups together with the carbon atom to which they are attached form a spiro oxetanyl group. In some embodiments, two R 8 groups together with the carbon atom to which they are attached form a fused heterocyclic group composed of 3 to 5 atoms. In some embodiments, two R 8 groups together with the carbon atom to which they are attached form a fused oxetanyl group. In some embodiments, two R 8 groups together with the carbon atom to which they are attached form a spiro or fused oxetanyl group. In some embodiments, the heterocyclic group contains 1 - 3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the heterocyclic group contains 1 nitrogen atom. In some embodiments, the heterocyclic group contains 2 nitrogen atoms. In some embodiments, the heterocyclic group contains 1 oxygen atom. In some embodiments, the heterocyclic group contains 2 oxygen atoms. In some embodiments, the heterocyclic group contains 1 oxygen atom and 1 nitrogen atom. In some embodiments, the heterocyclic group contains 1 sulfur atom. In some embodiments, the heterocyclic group contains 1 nitrogen atom and 1 sulfur atom. In some embodiments, two R 8 groups together with the carbon atom to which they are attached form a spiro aziridinyl, spiro oxiranyl, spiro oxetanyl, spiro azetidinyl, spiro tetrahydrofuranyl, spiro dioxolanyl, spiro pyrrolidinyl, spiro pyrazolyl, or spiro isoxazolyl group. In some embodiments, two R 8 groups together with the carbon atom to which they are attached form a fused aziridinyl, fused oxiranyl, fused oxetanyl, fused azetidinyl, fused tetrahydrofuranyl, fused dioxolanyl, fused pyrrolidinyl, fused pyrazolyl, or fused isoxazolyl group.

[0215] In some embodiments, X is In some embodiments, X is In some embodiments, X is In some embodiments, X is In some embodiments, X is In some embodiments, X is In any of these embodiments, two R 7 groups can both be hydrogen (H). In any of these embodiments, one R 7 group can be H and the other R 7 group can be -CH 3 . In any of these embodiments, two R 7 groups can both be -CH 3 .

[0216] In some embodiments, the compound has a structure represented by formula (I-A) or (I-B):

[0217]

[0218] wherein R 1 , R 2 , R 3 , R 4 , R 5 , Z 1 , Z 2 and X are as described for the compound of formula (I).

[0219] In some embodiments, the compound has a structure represented by formula (I-a) or (I-b):

[0220]

[0221] wherein R 1 , R 2 , R 5 , R 6 , Z 1 , Z 2 and X are as described for the compound of formula (I).

[0222] In some embodiments, the compound has a structure represented by formula (I-C), (I-D), (I-E), (I-F), (I-G), (I-H), or (I-J):

[0223]

[0224]

[0225] wherein R 3 , R 4 , R 5 and X are as described for the compound of formula (I).

[0226] 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), or (II-H):

[0227]

[0228] wherein R 3 、R 4 、R 5 、R 7 、R 8 and the ring B group moiety are as described for the compounds of formula (I).

[0229] In some embodiments, the compound has a structure represented by formula (III-A), (III-B), (III-C), (III-D), (III-E), (III-F), (III-G), or (III-H):

[0230]

[0231] wherein R 3 、R 4 、R 5 、R 7 、R 8 and Y are as described for the compounds of formula (I).

[0232] In some embodiments, the compound has a structure represented by formula (IV-A), (IV-B), (IV-C), (IV-D), (IV-E), (IV-F), (IV-G), or (IV-H):

[0233]

[0234]

[0235] wherein R 3 、R 4 and R 5 are as described for the compounds of formula (I), and X is H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkyl-OH, C 1 -C 6 alkyl-CN, C 8 optionally substituted with 1-5 R 3 -C 6 cycloalkyl. In some embodiments, X is H. In some embodiments, X is C 1 -C 6 alkyl. In some embodiments, X is C 1 -C 6Halogenated alkyl. In some embodiments, X is C 1 -C 6 alkyl-OH. In some embodiments, X is C 1 -C 6 alkyl-CN. In some embodiments, X is optionally substituted with 1-5 R 8 groups and is C 3 -C 6 cycloalkyl.

[0236] Table 1. Representative Compounds of the Invention

[0237]

[0238]

[0239]

[0240]

[0241]

[0242] In some embodiments, the present invention provides compounds selected from Compound Nos. 1-53 in Table 1, or their tautomers, or their pharmaceutically acceptable salts.

[0243] On the one hand, the present invention provides a method for preparing the compound represented by formula (I) described in the present invention. On the other hand, the present invention provides intermediate compounds for preparing the compound represented by formula (I). The present invention also provides compounds used as assay probes and optionally labeled with, for example, fluorescent labels. On the other hand, the present invention provides a method for assaying Cbl-b inhibition. In one variant, the present invention provides a method for assaying Cbl-b inhibition, which comprises pre-incubating Cbl-b with an assay probe (e.g., an assay probe labeled with a fluorescent label), then exposing the Cbl-b / assay probe mixture to a candidate compound, and then using, for example, FRET signal detection to determine whether the assay probe is displaced by the candidate compound and the degree of displacement.

[0244] The following scheme describes a method for synthesizing the compounds disclosed in the present invention. A mixture of stereoisomers generated during the synthesis, such as a racemic mixture of the final compound, can be separated into their respective enantiomers using common chromatographic methods, such as supercritical fluid chromatography with a chiral stationary phase, chiral column chromatography, or other methods known in the art.

[0245] Scheme I.

[0246]

[0247] Intermediate compounds of general formula I-5 or I-6 can be synthesized as described in Scheme I, where R 3 and R 4 are both as defined for the compounds of formula (I), and Y 1 is H to form a suitable ester, or is another suitable carboxylic acid protecting group. The groups R 3 and R 4 can be installed by deprotonating an arylacetic acid (such as I-1) with a base (such as NaH, i-PrMgCl, KO-t-Bu or LHMDS), followed by treatment with an electrophile (such as an alkyl bromide or formaldehyde), or by using a di-electrophile (such as 2-methyl-1,3-dibromopropane or epichlorohydrin) (to provide compounds in which R 3 and R 4 together with the carbon atom to which they are attached form a cycloalkyl group). Certain compounds of formula I-2, such as those in which R 3 and R 4 together with the carbon atom to which they are attached form a cyclopentyl group, are available directly from commercial sources. Compounds of formula I-2 in which R 3 and R 4 are each hydroxymethyl form an oxetane ring by Mitsunobu cyclization. Triazole assembly can be achieved by hydrazide formation, cyclization and desulfurization to provide compounds of formula I-5. The subsequent conversion of I-5 to compounds of formula I-6 can be achieved by amination using ammonia and copper or a Boc carbamate and palladium, followed by deprotection.

[0248] Scheme II.

[0249]

[0250] Intermediate I-5 can be further refined as shown in Scheme II. When R 3 and R 4 form an oxo-substituted ring (I-5a), the ketone can be homologated to a cyano-olefin II-1, and the olefin can be reduced to a cyanomethyl-substituted intermediate II-2. When R 3 and R 4 form a hydroxy-substituted ring, the hydroxy group can be alkylated with an electrophile to give II-3, which can then be converted to a triazole II-4. The stereochemistry can be inverted by a Mitsunobu reaction, followed by a similar alkylation, to give intermediates such as II-7 and II-8. Further substitution can be achieved by converting the hydroxy group to a nitrile group by mesylation and displacement, to give II-10 and II-11.

[0251] Scheme III.

[0252]

[0253] wherein R 3 and R 4 Compounds in which, together with the carbon atom to which they are attached, form a tetrahydrofuran ring can be assembled as shown in Scheme III. Bromide III-1 can be coupled with activated dihydropyran under palladium catalysis to give alkene III-2. Alkene III-2 can be epoxidized and subjected to a Lewis acid-promoted ring contraction to form aldehyde III-3. Aldehyde III-3 is oxidized to the carboxylic acid, and then refined to triazole as outlined in Scheme I to give intermediate III-4.

[0254] Scheme IV.

[0255]

[0256] Scheme IV shows the assembly of compounds in which the ring A group moiety is an indolinone ring, wherein R 1 、R 3 、R 4 、R 5 、R 7 、R 8 、Z 1 and the ring B group moiety all have the definitions given for the compounds of formula (I). Formula IV-1 can be directly condensed with aniline I-6 to give intermediate IV-4. Then the compound of formula IV-4 is treated with a heterocycle containing the ring B group moiety under reductive amination conditions to provide a compound of general formula IV-5. Alternatively, the order of steps can be reversed, and IV-1 and a heterocycle containing the ring B group moiety are subjected to reductive amination to give intermediate IV-2. Then the intermediate of formula IV-2 can be condensed with aniline I-6 to provide a compound of general formula IV-5. In certain embodiments, the intermediate of formula IV-2 can be cyclized to form an indolinone compound IV-3, which is then coupled to bromide I-5 under palladium catalysis.

[0257] Scheme V.

[0258]

[0259] Scheme V shows the synthesis of intermediate compounds, where two R 7 groups are each independently a C 1 -C 6 alkyl derivative, and wherein R 1 、R 3 、R 4 、R 7 、R 8 、Z 1 and the ring B group moiety all have the definitions given for the compounds of formula (I). Using a strong dehydrating agent such as Ti(OEt) 4Condense a ketone of general formula V-1 with a heterocycle containing the ring B group moiety, then add a cyanide to provide a Strecker intermediate, such as V-2. A second R substituent can then be installed by adding the appropriate Grignard reagent to provide a quaternary compound V-3, which can then be coupled with bromide I-5 to provide a compound of formula V-4. 7 Substituents to provide a quaternary compound V-3, which can then be coupled with bromide I-5 to provide a compound of formula V-4.

[0260] Scheme VI.

[0261]

[0262] Scheme VI outlines the synthesis of compounds of general formula VI-5, where R, R, R, R, Z, and X all have the definitions given for the compounds of formula (I); R forms part of a suitable ester or is another suitable carboxylic acid protecting group; Y is an alkyl group such as methyl, a cycloalkyl group such as cyclopropyl, a haloalkyl group such as bromomethyl, or -CHO; and Y is a hydroxyl group or NH. A methylpyridine or pyrimidine compound of formula VI-1 is carried directly onto intermediate VI-4, where X is methyl, for coupling to an intermediate compound of formula I-5 or I-6. Alternatively, the methyl group can be activated by oxidation with SeO to provide an aldehyde, or with Br to form a bromomethyl derivative of formula VI-2. The amino group at X can then be installed by reductive amination or displacement with a substituted amine to provide compound VI-3, which is then subjected to ester hydrolysis under basic conditions to provide compound VI-4, where Y is OH. The acid VI-4 is then coupled with an amine I-6 using a coupling agent such as HATU or T3P to give an amide VI-5; amide VI-4 (where Y is NH) is coupled with bromide I-5 under palladium catalysis. 2 R, 3 R, 4 R, 5 R, 2 Z, and X all have the definitions given for the compounds of formula (I); R forms part of a suitable ester or is another suitable carboxylic acid protecting group; Y is an alkyl group such as methyl, a cycloalkyl group such as cyclopropyl, a haloalkyl group such as bromomethyl, or -CHO; and Y is a hydroxyl group or NH. A methylpyridine or pyrimidine compound of formula VI-1 is carried directly onto intermediate VI-4, where X is methyl, for coupling to an intermediate compound of formula I-5 or I-6. Alternatively, the methyl group can be activated by oxidation with SeO to provide an aldehyde, or with Br to form a bromomethyl derivative of formula VI-2. The amino group at X can then be installed by reductive amination or displacement with a substituted amine to provide compound VI-3, which is then subjected to ester hydrolysis under basic conditions to provide compound VI-4, where Y is OH. The acid VI-4 is then coupled with an amine I-6 using a coupling agent such as HATU or T3P to give an amide VI-5; amide VI-4 (where Y is NH) is coupled with bromide I-5 under palladium catalysis. b forms part of a suitable ester or is another suitable carboxylic acid 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 is a hydroxyl group or NH 2 . A methylpyridine or pyrimidine compound of formula VI-1 is carried directly onto intermediate VI-4, where X is methyl, for coupling to an intermediate compound of formula I-5 or I-6. Alternatively, the methyl group can be activated by oxidation with SeO to provide an aldehyde, or with Br to form a bromomethyl derivative of formula VI-2. The amino group at X can then be installed by reductive amination or displacement with a substituted amine to provide compound VI-3, which is then subjected to ester hydrolysis under basic conditions to provide compound VI-4, where Y 2 is OH. The acid VI-4 is then coupled with an amine I-6 using a coupling agent such as HATU or T3P to give an amide VI-5; amide VI-4 (where Y 2 is NH c is OH. Then acid VI-4 is coupled with an amine I-6 using a coupling agent such as HATU or T3P to give an amide VI-5; amide VI-4 (where Y c is NH 2 ) is coupled with bromide I-5 under palladium catalysis.

[0263] Compounds 1, 3, 4, 17, and 35 - 53 exhibited IC values of 5 nM or less in the Cbl-b inhibition assay of Biological Example 1 and are useful in a pharmaceutical composition and method disclosed in one embodiment of the present invention. Compounds 8, 15, and 25 - 34 exhibited IC values between greater than 5 nM and 20 nM in the Cbl-b inhibition assay of Biological Example 1. 50 values, and are useful in a pharmaceutical composition and method disclosed in one embodiment of the present invention. Compounds 8, 15, and 25 - 34 exhibited IC values between greater than 5 nM and 20 nM in the Cbl-b inhibition assay of Biological Example 1. 50values, and in one embodiment for the pharmaceutical compositions and methods disclosed herein. Compounds 5, 6, 14, 16, and 19 - 24 exhibited IC values between greater than 20 nM and 100 nM in the Cbl-b inhibition assay of Biological Example 1 50 values, and in one embodiment for the pharmaceutical compositions and methods disclosed herein. Compounds 2, 7, 9 - 13, and 18 exhibited IC values greater than 100 nM in the Cbl-b inhibition assay of Biological Example 1 50 values, and in one embodiment for the pharmaceutical compositions and methods disclosed herein.

[0264] In various embodiments, and as further described herein, as determined by the Cbl-b inhibition assay of Biological Example 1, the compounds provided by the present invention (as well as compositions comprising the compounds described herein, and methods of using the compounds or compositions described herein) have an IC value of 5 nM or less, between greater than 5 nM and 20 nM, between greater than 20 nM and 100 nM, or greater than 100 nM 50 value. In a further embodiment, and as further described herein, as determined by the Cbl-b inhibition assay of Biological Example 1, the compounds provided by the present invention (as well as compositions comprising the compounds described herein, and methods of using the compounds or compositions described herein) have an IC value of 5 nM or less 50 value. In a further embodiment, and as further described herein, as determined by the Cbl-b inhibition assay of Biological Example 1, the compounds provided by the present invention (as well as compositions comprising the compounds described herein, and methods of using the compounds or compositions described herein) have an IC value between greater than 5 nM and 20 nM 50 value. In a further embodiment, and as further described herein, as determined by the Cbl-b inhibition assay of Biological Example 1, the compounds provided by the present invention (as well as compositions comprising the compounds described herein, and methods of using the compounds or compositions described herein) have an IC value between greater than 20 nM and 100 nM 50 value. In a further embodiment, and as further described herein, as determined by the Cbl-b inhibition assay of Biological Example 1, the compounds provided by the present invention (as well as compositions comprising the compounds described herein, and methods of using the compounds or compositions described herein) have an IC value greater than 100 nM 50 value.

[0265] For the secretion of IL-2 by immune cells (e.g., T cells) co-stimulated with anti-CD3 antibody and anti-CD28 antibody, at an inhibitor concentration of 1 μM or 0.3 μM, the compounds provided by the present invention (and compositions comprising the compounds described in the present invention) induce a change less than 20-fold, between 20 - 35-fold, or greater than 35-fold relative to the baseline.

[0266] For the secretion of IL-2 by immune cells (e.g., T cells) stimulated with anti-CD3 antibody, at an inhibitor concentration of 3 μM or 1 μM, the compounds provided by the present invention (and compositions comprising the compounds described in the present invention) induce a change less than 0.70-fold, between 0.70 - 1.1-fold, or greater than 1.1-fold relative to the baseline.

[0267] For the CD25 staining on the cell surface of immune cells (e.g., T cells) co-stimulated with anti-CD3 antibody and anti-CD28 antibody, at an inhibitor concentration of 1 μM or 0.3 μM, the compounds provided by the present invention (and compositions comprising the compounds described in the present invention) induce a change less than 1.24-fold, between 1.24 - 1.39-fold, or greater than 1.39-fold relative to the baseline.

[0268] For the CD25 staining on the cell surface of immune cells (e.g., T cells) stimulated with anti-CD3 antibody, at an inhibitor concentration of 3 μM or 1 μM, the compounds provided by the present invention (and compositions comprising the compounds described in the present invention) induce a change less than 1.5-fold, between 1.5 - 2.5-fold, or greater than 2.5-fold relative to the baseline.

[0269] III. Uses and Methods

[0270] The present invention provides methods for modulating the activity of immune cells (e.g., T cells, B cells, or NK cells), such as by contacting the immune cells with an effective amount of a Cbl-b inhibitor or a composition thereof of the present invention. The present invention also provides in vitro methods for generating such immune cells with 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 suffering from cancer) by ex vivo methods. The present invention further provides in vivo methods for modulating responses in an individual in need (e.g., an individual suffering from cancer), wherein the method comprises administering an effective amount of a Cbl-b inhibitor or a composition thereof of the present invention. In addition, the present invention provides in vitro methods for generating an expanded population of lymphocytes following lymphomodulation in an individual, wherein the lymphomodulation occurs due to administration of an effective amount of a Cbl-b inhibitor or a composition thereof of the present invention to the individual. The expanded population of lymphocytes can then be administered to an individual suffering from cancer. In some embodiments, the modified immune cells or the expanded population of lymphocytes are generated 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 (TIL)).

[0271] In addition, the present invention provides Cbl-b inhibitors for use as therapeutic active substances. The present invention provides Cbl-b inhibitors for treating or preventing diseases or disorders 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 a Cbl-b inhibitor in the preparation of a medicament for treating or preventing a disease or disorder associated with Cbl-b activity. The present invention also provides the use of a Cbl-b inhibitor in the preparation of a medicament for treating cancer. In addition, the present invention provides treatment methods, medicaments, and uses comprising a Cbl-b inhibitor as part of a combination therapy for treating cancer, the combination therapy for treating cancer involving one or more of immune checkpoint inhibitors, anti-tumor agents, and radiotherapy.

[0272] In some embodiments of the treatment methods, medicaments, and uses of the present invention, the cancer is a hematological cancer, such as lymphoma, leukemia, or myeloma. In other embodiments of the treatment methods, medicaments, and uses of the present invention, the cancer is a non-hematological cancer, such as sarcoma, carcinoma, or melanoma.

[0273] Hematological cancers include, but are not limited to, one or more leukemias, such as B-cell acute lymphoblastic leukemia (“B-ALL”), T-cell acute lymphoblastic leukemia (“T-ALL”), acute lymphoblastic leukemia (ALL); one or more chronic leukemias, including but not limited to chronic myelogenous 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, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, follicular small cell lymphoma or follicular large cell lymphoma, malignant lymphoproliferative disease, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndromes, non-Hodgkin lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom macroglobulinemia, and “preleukemia”, which are all various hematological diseases united by ineffective production (or dysplasia) of myeloid blood cells.

[0274] 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.

[0275] In some cases, the effectiveness of administering a Cbl-b inhibitor in treating a disease or disorder (such as cancer) is determined by evaluating clinical outcomes (such as reduction in tumor size or number of tumors, and / or survival rate). In some embodiments, “treating cancer” includes evaluating the patient's response to a treatment regimen according to the Response Evaluation Criteria in Solid Tumors (RECIST), version 1.1, as described (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 efficacy evaluation criteria for determining objective anti-tumor response according to RECIST 1.1 include: complete response (CR); partial response (PR); progressive disease (PD); and stable disease (SD).

[0276] A. Isolation and treatment of cells

[0277] 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 cell" refers to an immune cell or a cell population comprising the immune cell that has been cultured, incubated, and / or contacted with an effective amount of a Cbl-b inhibitor to modulate the activity of the immune cell. In some embodiments, the modified immune cells can be used in immunotherapy, e.g., in combination with adoptive immunotherapy methods.

[0278] 1. Sample

[0279] In some embodiments, the immune cells to be modified or the cell population comprising the immune cells to be modified are isolated from a sample, such as a biological sample, e.g., a sample obtained or derived from an individual (e.g., a human), such as a biological sample. In some embodiments, the individual from whom the immune cells are isolated is an individual suffering from a specific disease or disorder (e.g., cancer) or in need of or to be administered cell therapy. In some embodiments, the individual is a human in need of a specific therapeutic intervention, e.g., adoptive cell therapy, wherein immune cells are isolated, processed, and / or modified. Thus, in some embodiments, the cells isolated from the individual are primary cells (e.g., primary human cells). As used herein, the term "primary cell" refers to a cell isolated directly from a mammalian biological fluid or tissue (e.g., a human biological fluid or tissue).

[0280] 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 herein, the term "hematopoietic cell" includes hematopoietic stem cells and hematopoietic progenitor cells. In some embodiments, the immune cells to be modified are present in a heterogeneous cell population or a composition comprising a heterogeneous cell population. For example, the immune cells to be modified can be hematopoietic cells present in a heterogeneous cell population that comprises 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 population or a composition comprising a homogeneous cell population. For example, the immune cells to be modified can be hematopoietic cells present in a homogeneous cell population consisting only of hematopoietic cells. In some embodiments, the immune cells to be modified or the cell population comprising the immune cells to be modified comprise one or more subsets of immune cells. For example, one or more subsets of immune cells can be CD4+ cells, CD8+ cells, and their subsets, such as those defined by function, activation status, maturity, differentiation potential, expansion, localization, persistence, surface marker profile, cytokine secretion profile, and / or degree of differentiation.

[0281] In some embodiments, the biological samples of the present invention include tissues, body fluids, and other samples directly taken from an individual, as well as samples generated 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 a tissue or organ containing a tumor), 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.

[0282] In some cases, the biological sample from which the immune cells are derived or isolated is blood or a blood-derived sample, or a product derived from apheresis or leukapheresis.

[0283] Exemplary biological samples include whole blood, peripheral blood mononuclear cells (PBMCs), white blood cells, bone marrow, thymus, tissue biopsy, tumor, leukemia, lymphoma, lymph node, gut-associated lymphoid tissue, mucosa-associated lymphoid tissue, spleen, other lymphoid tissues, liver, lung, stomach, intestine, colon, kidney, pancreas, breast, bone, prostate, cervix, testis, 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 the 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).

[0284] 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, such as from a mouse, a rat, a non-human primate, or a pig.

[0285] 2. Cell Processing and Separation

[0286] In some embodiments, the isolation of the immune cells to be modified includes one or more preparation and / or cell separation steps. The one or more cell separation steps can be non-affinity-based separation or affinity-based separation. As an example, non-affinity-based separation can be centrifugation of a composition comprising the immune cells to be modified. In some embodiments, the non-affinity-based separation method includes density-based cell separation methods, such as preparing white blood cells from peripheral blood by lysing red blood cells and centrifuging through a Percoll or Ficoll gradient. Affinity-based separation methods can include contacting a composition comprising the 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., sold by Miltenyi Biotec Inc., Auburn, CA microbeads; or EasySep sold by Stemcell Technologies, Vancouver, BC, Canada TM Direct RapidSpheres TM ), the antibody binding to a marker expressed on the surface of the immune cells to be modified. In some embodiments, specific subsets 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 techniques in which target cells (e.g., the immune cells to be modified) bind to a reagent and are retained for further use. For example, magnetic beads conjugated with anti-CD3 antibody can be used (e.g., CD3 microbeads) perform positive selection on CD3+ T cells. Negative selection can be based on techniques that retain target cells that have not bound to the reagent (e.g., immune cells to be modified). For example, negative selection can be used to isolate total human primary T cells from peripheral blood mononuclear cells (PMBCs), where a mixture of antibodies against surface markers CD14, CD15, CD16, CD19, CD34, CD36, CD56, CD123, and CD235a is incubated in a sample containing PBMCs, and then the sample is passed over magnetic beads to remove cells expressing those surface markers and retain the remaining cells in the sample for further processing. In some embodiments, the immune cells or cell population containing the immune cells to be modified are washed, centrifuged, and / or incubated in the presence of one or more reagents, e.g., to remove unwanted components, enrich desired components, and / or lyse or remove cells sensitive to a particular reagent. In some examples, the immune cells are isolated based on one or more characteristics (e.g., density, adhesion properties, size, sensitivity, and / or resistance to a particular component). The cell isolation step does not require 100% enrichment or removal of a particular cell. In some embodiments, positive selection or enrichment of a particular type of immune cell (e.g., CD4+ T cells) refers to increasing the number or percentage of such cells. In some embodiments, removal or depletion of a particular type of cell that is not of interest, e.g., by negative selection, refers to reducing the number or percentage of such cells.

[0287] In some embodiments, the immune cells or cell population containing the immune cells are obtained from the circulating blood of an individual, e.g., by apheresis or leukapheresis. In some cases, a sample containing 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.

[0288] In some embodiments, the blood cells collected from an individual are washed, e.g., to remove the plasma fraction and place the cell population containing the immune cells to be modified in a suitable buffer or medium for subsequent processing steps. In some embodiments, the cell population containing the immune cells to be modified is washed with phosphate-buffered saline. In some embodiments, the washing solution is calcium- and / or magnesium-free. In some cases, the washing step is performed using a semi-automatic "flow-through" centrifuge. In some cases, the washing step is performed 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, e.g., calcium- and / or magnesium-free phosphate-buffered saline, after washing. In some embodiments, the components of the blood cell sample are removed and the immune cells to be modified or the cell population containing the immune cells to be modified are directly resuspended in a suitable cell culture medium.

[0289] Representative methods for processing and / or isolating immune cells (e.g., hematopoietic cells) from a sample, which sample comprises a cell population containing the hematopoietic cells (e.g., a sample comprising PBMC), are described in Biological Example 2 and Biological Example 3 of the present invention. Methods and techniques for processing and / or isolating 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.

[0290] 3. Incubation and Treatment

[0291] The present invention provides a method for modulating the activity of immune cells (such as the processed and / or isolated immune cells described above) by contacting the immune cells with an effective amount of a Cbl-b inhibitor of the present invention. The present invention also provides modified immune cells generated by any method of the present invention, for example, by culturing a cell population containing immune cells (e.g., the processed and / or isolated immune cells described above) in the presence of an effective amount of a Cbl-b inhibitor to modulate the activity of the immune cells, thereby generating the modified immune cells.

[0292] In some embodiments, the immune cells to be modified (e.g., the processed and / or isolated immune cells described above) are incubated and / or cultured in a suitable medium before contacting the immune cells with the Cbl-b inhibitor provided by the present invention. In some embodiments, the immune cells to be modified are incubated and / or cultured in a suitable medium to simultaneously contact the immune cells with the Cbl-b inhibitor provided by the present invention.

[0293] The processed and / or separated immunocytes to be modified or a cell population comprising the immunocytes to be modified can be differentiated and / or expanded in vitro. In some embodiments, the immunocytes 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 immunocytes to be modified are incubated in a suitable cell culture medium comprising the Cbl-b inhibitor of the present invention before the immunocyte differentiation and / or expansion. In some embodiments, the immunocytes to be modified are incubated in a suitable cell culture medium comprising the Cbl-b inhibitor of the present invention after the immunocyte differentiation and / or expansion. The immunocytes are modified (i.e., modified immunocytes) when contacted with an effective amount of the Cbl-b inhibitor provided by the present invention to modulate the immunocyte activity. In some embodiments, the immunocytes to be modified are not differentiated and / or expanded in vitro, and thus are the same cell type as the modified immunocytes that have been contacted with the Cbl-b inhibitor. For example, T cells can be cultured in a suitable medium comprising a Cbl-b inhibitor without differentiating the T cells. In other embodiments, the immunocytes to be modified are differentiated and / or expanded in vitro, and thus are different cell types from the modified immunocytes that have been contacted with the Cbl-b inhibitor. For example, hematopoietic cells can be incubated in a suitable medium comprising a Cbl-b inhibitor and other agents that can drive the differentiation of the hematopoietic cells into mature hematopoietic cells. Thus, in some cases of the embodiments of the present invention, the modified immunocytes are hematopoietic cells, pluripotent stem cells, myeloid progenitor cells, lymphoid progenitor cells, T cells, B cells, and / or NK cells. Methods for the expansion and / or differentiation of immunocytes are well known in the art. See, for example, International Patent Application No. WO 2017 / 037083.

[0294] An effective amount of a Cbl-b inhibitor is the amount or concentration of a Cbl-b inhibitor that is sufficient to modulate the activity of immune cells as compared to a reference sample. The reference sample can be immune cells that have not been contacted with a Cbl-b inhibitor. In some embodiments, the concentration of the Cbl-b inhibitor added to a composition (e.g., a 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 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 a composition (e.g., a cell culture medium) comprising 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 10 nM, about 20 nM, about 40 nM, about 50 nM, about 80 nM, about 0.1 μM, about 0.2 μM, about 0.3 μM, about 0.4 μM, about 0.5 μM, about 1 μM, about 5 μM, about 10 μM, about 15 μM, about 20 μM, about 25 μM, about 30 μM, about 40 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, about 90 μM, or about 100 μM. In some embodiments, the concentration of the Cbl-b inhibitor added to a composition (e.g., a 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 a composition (e.g., a cell culture medium) comprising the immune cells to be modified is about 1 μM or about 8 μM.

[0295] Compared with a reference sample, an effective amount of a Cbl-b inhibitor contacts immune cells for a sufficient length of time to modulate the activity of the immune cells. The reference sample can be immune cells that have not been contacted with the Cbl-b inhibitor but have been incubated for the same length of time in a composition (e.g., cell culture medium) containing the immune cells and the Cbl-b inhibitor. In some embodiments, the Cbl-b inhibitor contacts and / or incubates 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 contacts and / or incubates 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 contacts and / or incubates 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 contacts and / or incubates 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 contacts and / or incubates 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.

[0296] 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, one or more of 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).

[0297] In some embodiments, suitable conditions for inducing the proliferation, expansion, activation, and / or survival of immune cells include providing stimulatory conditions comprising an agent capable of activating the immune cells (e.g., NK cells). For example, suitable conditions for inducing the proliferation, expansion, activation, and / or survival of T cells include providing stimulatory conditions capable of activating intracellular signaling in T cells. Complete activation of T cells generally requires recognition of an antigen by the T cell receptor (referred to herein as "TCR" (signal 1)) and recognition of a co-stimulator such as CD28 (signal 2). In some cases, one or more agents initiate or activate the intracellular signaling cascade mediated by the TCR complex in T cells. For example, a first agent may bind to a component of the TCR complex to activate the T cell, and a second agent may bind to a co-stimulatory molecule on the surface of the T cell to stimulate the activated T cell. In some embodiments, the first agent stimulates TCR / CD3 complex-associated signals in T cells by specifically binding to CD3 (e.g., anti-CD3 antibody). In further embodiments, the co-stimulatory molecule on the surface of the T cell may be CD28, and the second agent specifically binds to CD28 (e.g., anti-CD28 antibody). Such agents include, but are not limited to, antibodies, bivalent antibody fragments, and binding molecules, such as those specific for components of the TCR complex (e.g., anti-CD3 antibody) and / or those specific for co-stimulatory receptors (e.g., anti-CD28 antibody). In some embodiments, the agent that specifically binds to CD3 is an anti-CD3 antibody, a bivalent antibody fragment of an anti-CD3 antibody (e.g., (Fab)2′ fragment or bivalent scFv fragment), a monovalent antibody fragment of an anti-CD3 antibody (e.g., Fab fragment, Fv fragment, or scFv fragment), or a CD3-binding molecule (e.g., 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., (Fab)2′ fragment or bivalent scFv fragment), a monovalent antibody fragment of an anti-CD28 antibody (e.g., Fab fragment, Fv fragment, or scFv fragment), and a CD28-binding molecule (e.g., aptamer). For example, one or more agents provided by the present invention (e.g., anti-CD3 antibody and anti-CD28 antibody) may be bound to a solid support (e.g., beads) or crosslinked with an anti-Fc antibody. In some embodiments, the amplification method step may further comprise the step of adding an anti-CD3 antibody and / or an anti-CD28 antibody to the culture medium. In some embodiments, the stimulant 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 may be added to the cell culture medium containing the immune cells and the agent (e.g., anti-CD3 antibody and / or anti-CD28 antibody) at a concentration of at least about 10 units / mL.

[0298] In some embodiments, suitable conditions for inducing T cell proliferation, expansion, activation, and / or survival include providing stimulatory conditions or agents capable of activating intracellular signaling through the T cell receptor (TCR) complex, as well as a Cbl-b inhibitor as described herein. In some embodiments, the immune cells or cell population comprising the immune cells are incubated with a first agent (e.g., an anti-CD3 antibody) that stimulates TCR / CD3 complex-associated signaling in T cells by specifically binding to CD3. In further embodiments, the immune cells or cell population comprising the immune cells are incubated with a first agent (e.g., an anti-CD3 antibody) that stimulates TCR / CD3 complex-associated signaling in T cells by specifically binding to CD3, a second agent (e.g., an anti-CD28 antibody) that binds to the co-stimulatory molecule CD28, and a Cbl-b inhibitor at 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 in the presence of a Cbl-b inhibitor do not require stimulation through co-stimulatory molecules (e.g., CD28). Contacting T cells with a Cbl-b inhibitor or a composition thereof can bypass the need for co-stimulation required for T cells to enter an activated state. In certain embodiments, the immune cells or cell population comprising the immune cells are incubated with a first agent (e.g., an anti-CD3 antibody) that stimulates TCR / CD3 complex-associated signaling in T cells by specifically binding to CD3, and a Cbl-b inhibitor at 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).

[0299] In some embodiments of the method of modulating the activity of immune cells, the immune cells are T cells, and modulating the activity of T cells comprises increased T cell activation and / or increased T cell proliferation. Even in the presence of an activator that binds to a component of the TCR complex (e.g., anti-CD3 antibody), and in the presence of a stimulator that binds to a co-stimulatory molecule (e.g., anti-CD28 antibody), the T cells encompassed by the embodiments of the present invention can be in a tolerant state. In some embodiments, the method of modulating the activity of T cells comprises contacting T cells with an effective amount of a Cbl-b inhibitor or a composition thereof in the presence of a combination of anti-CD3 antibody and anti-CD28 antibody. In some embodiments, the method of modulating the activity of T cells 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 a combination of anti-CD3 antibody and anti-CD28 antibody. In some embodiments, modulating the activity of T cells (e.g., increasing T cell activation and / or increasing T cell proliferation) does not require stimulation via the co-stimulatory CD28 molecule. In some embodiments, the method of modulating the activity of T cells comprises contacting T cells with an effective amount of a Cbl-b inhibitor or a composition thereof in the sole presence of an anti-CD3 antibody. In some embodiments, the method of modulating the activity of T cells 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 antibody), and wherein the agent does not include an agent that stimulates the CD28 co-stimulatory molecule (e.g., anti-CD28 antibody).

[0300] In some embodiments, the immune cells are T cells, and modulating T cell activity comprises enhanced T cell activation and / or enhanced T cell proliferation. For example, such as in the presence of an agent that activates T cells (e.g., anti-CD3 antibody), and in some further embodiments, in the presence of an agent that stimulates T cells (e.g., anti-CD28 antibody), the T cells encompassed by the embodiments of the present invention can be in an activated state. Contacting T cells with a Cbl-b inhibitor or a composition thereof can lower the threshold required for activation, and thus, in the presence of an activator (e.g., anti-CD3 antibody) and in some further embodiments in the presence of a stimulator (e.g., anti-CD28 antibody), can enhance T cell activation and / or proliferation. In some embodiments, a method of modulating T cell activity comprises contacting T cells with an effective amount of a Cbl-b inhibitor or a composition thereof in the presence of a combination of anti-CD3 antibody and anti-CD28 antibody. In some embodiments, a method of modulating T cell activity comprises contacting T cells with an effective amount of a Cbl-b inhibitor or a composition thereof, wherein the T cells have been previously contacted with a combination of anti-CD3 antibody and anti-CD28 antibody. In some embodiments, modulating T cell activity (e.g., enhancing T cell activation and / or enhancing T cell proliferation) does not require stimulation via the co-stimulatory CD28 molecule. In some embodiments, a method of modulating T cell activity comprises contacting T cells with an effective amount of a Cbl-b inhibitor or a composition thereof in the sole presence of anti-CD3 antibody. In some embodiments, a method of modulating T cell activity comprises contacting T cells with an effective amount of a Cbl-b inhibitor or a composition thereof, wherein the T cells have been previously contacted with one or more agents that activate T cells (e.g., anti-CD3 antibody).

[0301] In some embodiments, the immune cells are T cells, and modulating T cell activity involves reducing T cell dysfunction, including reduced T cell exhaustion, decreased T cell tolerance, and / or reduced T cell anergy. The general principles of T cell dysfunction are well known in the art (see, e.g., 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 a reduced responsiveness to an antigen due to prior exposure to the antigen. When a specific lymphocyte (such as a T cell) encounters an antigen, the lymphocyte may be activated, leading to an antigen-specific immune response, or the lymphocyte (such as a T cell) may be inactivated or eliminated, resulting in antigen-specific immune tolerance. In some cases, tolerance can be caused by clonal anergy, peripheral clonal deletion, T cell suppression, and / or other forms of antigen-specific tolerance. In some embodiments, tolerance can be induced by or characterized by the induction of anergy. In some cases, anergy can be caused by exposing T cells to an antigen in the absence of costimulation. Prolonged antigen recognition solely through the TCR in the absence of a costimulatory signal can lead to anergy (i.e., functional unresponsiveness). Anergic T cells may be resistant to subsequent antigen challenges and may be able to suppress other immune responses. Generally, in the natural setting, tolerance involves non-responsiveness or non-generation of responses to self-antigens. However, in certain cases, tolerance to "non-self" antigens can be induced. Thus, in some cases, the same mechanisms by which mature T cells that recognize self-antigens in peripheral tissues fail to subsequently respond to these antigens may also regulate non-responsiveness to foreign or "non-self" antigens (e.g., antigens expressed by cancer cells). Thus, even in the presence of stimulants (e.g., agents that bind to costimulatory molecules such as CD28), the T cells encompassed by the embodiments of the present invention may be in a tolerant state. Contacting T cells with the Cbl-b inhibitor or its composition provided by the present invention can bypass certain aspects of T cell dysfunction (such as T cell tolerance, T cell anergy, and / or T cell exhaustion). In some embodiments, a method of modulating T cell activity (e.g., reducing T cell tolerance, reducing T cell anergy, and / or reducing T cell exhaustion) comprises contacting T cells with an effective amount of a Cbl-b inhibitor or its composition. In some embodiments of the methods of the present invention, modulating T cell activity (e.g., reducing T cell tolerance, reducing T cell anergy, and / or reducing T cell exhaustion) comprises contacting T cells with an effective amount of a Cbl-b inhibitor or its composition in the presence of a combination of anti-CD3 antibody and anti-CD28 antibody.In some embodiments of the method of the present invention, a method of modulating T cell activity (e.g., reducing T cell tolerance, reducing T cell anergy, and / or reducing T cell exhaustion) 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 a combination of an anti-CD3 antibody and an anti-CD28 antibody. In some embodiments, modulating T cell activity (e.g., reducing T cell tolerance, reducing T cell anergy, and / or reducing T cell exhaustion) does not require stimulation via the co-stimulatory CD28 molecule. In some embodiments of the method of the present invention, a method of modulating T cell activity (e.g., reducing T cell tolerance, reducing T cell anergy, 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 an anti-CD3 antibody alone. In some embodiments, a method of modulating T cell activity (e.g., reducing T cell tolerance, reducing T cell anergy, and / or reducing T cell exhaustion) 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., an anti-CD3 antibody alone).

[0302] Both T cell activation and T cell tolerance are tightly controlled processes that regulate the immune response. Accordingly, the present invention provides methods of modulating T cell activity, where modulating T cell activity comprises increased T cell activation, increased T cell proliferation, decreased T cell exhaustion, and / or decreased T cell tolerance. In some embodiments, the method of modulating T cell activity (e.g., increased T cell activation, increased T cell proliferation, decreased T cell exhaustion, and / or decreased T cell tolerance) comprises contacting the 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., increasing T cell activation, increasing T cell proliferation, decreasing T cell exhaustion, and / or decreasing T cell tolerance) comprises contacting the T cells with an effective amount of a Cbl-b inhibitor or a composition thereof in the presence of a combination of anti-CD3 antibody and anti-CD28 antibody. In some embodiments of the methods of the present invention, the 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 the T cells with an effective amount of a Cbl-b inhibitor or a composition thereof, wherein the T cells have been previously contacted with a combination of anti-CD3 antibody and anti-CD28 antibody. In some embodiments, modulating T cell activity (e.g., increasing T cell activation, increasing T cell proliferation, decreasing T cell exhaustion, and / or decreasing T cell tolerance) does not require stimulation via the co-stimulatory CD28 molecule. In some embodiments of the methods of the present invention, the 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 the T cells with an effective amount of the Cbl-b inhibitor or a composition thereof provided by the present invention in the presence of anti-CD3 antibody alone. In some embodiments, the 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 the T cells with an effective amount of a Cbl-b inhibitor or a composition thereof, wherein the T cells have been previously contacted with one or more agents that activate T cells (e.g., anti-CD3 antibody).

[0303] In some embodiments of the method of the present invention, increased T cell activation comprises increasing the production of one or more cytokines from T cells or surrounding immune cells (e.g., myeloid cells) in the microenvironment of the activated T cells. 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. Increased cytokine expression can be measured by ELISA.

[0304] In some embodiments of the method of the present invention, 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 T cell activation marker is one or more of the following: CD25, CD69, and CTLA4. Increased expression of cell surface markers can be measured by FACS.

[0305] Methods for experimentally determining increased T cell activation, increased T cell proliferation, decreased T cell exhaustion, and / or reduced T cell tolerance are well known in the art. In some embodiments, a representative method for determining T cell activation can be found in Biological Example 2 provided by the present invention. 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 reduced T cell tolerance can be found in Biological Example 3 provided by the present invention.

[0306] In some embodiments of the methods of modulating the activity of immune cells, the immune cells are B cells, and modulating the activity of B cells comprises increased B cell activation. In some embodiments, increased B cell activation comprises increased 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 major histocompatibility complex class II (e.g., HLA-DR). In some embodiments, the B cell activation marker is CD69. The increased expression of cell surface markers can be measured by FACS. In some embodiments, increased B cell activation comprises increased activation of proteins in signaling pathways (e.g., those mediated by ERK, JNK, and Syk). The increased activation of the proteins can be detected by measuring the phosphorylation levels on the proteins using reagents (e.g., anti-phosphorylation antibodies available in the art).

[0307] In some embodiments of the methods of modulating the activity of immune cells, the immune cells are NK cells, and modulating the activity of NK cells comprises increased NK cell activation. In some embodiments, increased NK cell activation comprises 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, increased NK cell activation comprises increased 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, increased NK cell activation includes increased killing of target cells (e.g., tumor cells, including primary tumor cells, and cell line-derived tumor cells, such as the K562 cell line).

[0308] Methods for experimental determination of 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., 6:252 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 learned from Biological Example 3 provided in the present invention. In some embodiments, representative methods for determining NK cell activation can be learned from Biological Example 3 provided in the present invention.

[0309] The activity regulation of immune cells (e.g., T cells, B cells, or NK cells) can be determined by measuring the baseline value of a 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 in contact with a Cbl-b inhibitor, can be measured before contacting or administering the Cbl-b inhibitor to determine the baseline value. Then a reference value of T cell activation is obtained after contacting or administering 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 administering the Cbl-b inhibitor or its composition. For example, in some embodiments, the activation of immune cells (e.g., T cells) in the sample increases by at least 0.1-fold compared to the baseline value, wherein the baseline value is obtained before contacting the immune cells (e.g., T cells) with the Cbl-b inhibitor or its composition. In some embodiments, the activation of immune cells (e.g., T cells) increases by at least about 0.1-fold, about 0.2-fold, about 0.3-fold, about 0.4-fold, about 0.5-fold, about 0.6-fold, about 0.7-fold, about 0.8-fold, about 0.9-fold, about 1-fold, about 2-fold, about 4-fold, about 6-fold, about 8-fold, about 10-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 75-fold, or about 100-fold (e.g., about 0.1-fold to about 100-fold, or about 1-fold to about 100-fold) compared to the baseline value. The activation of immune cells can be evaluated by measuring an increase in activation biomarkers such as an increase in cytokine secretion, an increase in cell surface expression of activation markers (e.g., cell surface markers), or an increase in phosphorylation of proteins in downstream signaling pathways. The fold change relative to the baseline value indicating immune cell activation can be determined for the parameter being tested and the conditions under which the immune cells are treated. For example, to measure T cell activation, the baseline value can be obtained from T cells stimulated with a combination of anti-CD3 antibody and anti-CD28 antibody, wherein the cells are not incubated with the Cbl-b inhibitor. Then a reference value is obtained from T cells stimulated with a combination of anti-CD3 antibody and anti-CD28 antibody, wherein the T cells have been in contact with or contacted with the Cbl-b inhibitor. Then a positive response of immune cell activation can be determined by the obtained reference value. Similar reference value measurements can be obtained and compared with the baseline values used to evaluate T cell activation, T cell proliferation, T cell exhaustion, T cell tolerance, B cell activation, and / or NK cell activation. The measurement values of these parameters can be obtained using techniques well known in the art and the techniques provided in Biological Example 2 and Biological Example 3.

[0310] As used herein, the term "baseline" or "baseline value" may refer to a measured value or characterization before administration of a therapeutic agent disclosed herein (e.g., a composition comprising a Cbl-b inhibitor as described herein) or at the start of administration of the therapeutic agent. The baseline value may 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 reduced T cell tolerance). As used herein, the term "reference" or "reference value" may refer to a measured value or characterization after administration of a therapeutic agent disclosed herein (e.g., a composition comprising a Cbl-b inhibitor as described herein). The reference value may be measured one or more times during an experimental time course, dose regimen, or treatment cycle or at the completion of an experimental time course, dose regimen, or treatment cycle. A "reference value" may be an absolute value, a relative value, a value with an upper and / or lower limit, a range of values, an average, a median, a mean, or a value compared to the baseline value. Similarly, a "baseline value" may be an absolute value, a relative value, a value with an upper and / or lower limit, a range of values, an average, a median, a mean, or a value compared to the reference value. The reference value and / or the baseline value may 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 set of samples (e.g., samples obtained from a set of two, three, four, five, or more individuals).

[0311] In some embodiments, the 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 antibody and anti-CD28 antibody in the presence of a Cbl-b inhibitor is at least 2.5-fold the baseline value of cytokine secretion (e.g., IL-2 secretion) obtained from T cells stimulated with a combination of anti-CD3 antibody and anti-CD28 antibody in the absence of a Cbl-b inhibitor. In some embodiments, the positive response to T cell activation as measured by surface marker expression (e.g., CD25 surface marker staining) of T cells stimulated with a combination of anti-CD3 antibody and anti-CD28 antibody in the presence of a Cbl-b inhibitor is at least 1.3-fold the baseline value of surface marker expression (e.g., CD25 surface marker staining) obtained from T cells stimulated with a combination of anti-CD3 antibody and anti-CD28 antibody in the absence of a Cbl-b inhibitor. In some embodiments, the baseline value can be obtained from T cells stimulated with anti-CD3 antibody alone, wherein the cells are not incubated with a Cbl-b inhibitor. In some embodiments, the positive response to T cell activation as measured by cytokine secretion (e.g., IL-2 secretion) of T cells stimulated with anti-CD3 antibody alone in the presence of a Cbl-b inhibitor is at least 0.1-fold the baseline value of cytokine secretion (e.g., IL-2 secretion) obtained from T cells stimulated with anti-CD3 antibody alone in the absence of a Cbl-b inhibitor. In some embodiments, the positive response to T cell activation as measured by surface marker expression (e.g., CD25 surface marker staining) of T cells stimulated with anti-CD3 antibody alone in the presence of a Cbl-b inhibitor is at least 0.6-fold the baseline value of surface marker expression (e.g., CD25 surface marker staining) obtained from T cells stimulated with anti-CD3 antibody alone in the absence of a Cbl-b inhibitor.

[0312] In some cases, the present invention provides a method for generating modified immune cells, which comprises culturing a cell population containing immune cells in the presence of an effective amount of a Cbl-b inhibitor or a composition thereof provided by the present invention to regulate the activity of immune cells, thereby generating the modified immune cells. In some embodiments, the immune cells are T cells, B cells, or natural killer (NK) cells.

[0313] In some embodiments of methods 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 a stimulant such as a cytokine or an antibody that binds to an activating protein expressed by the immune cells (e.g., anti-CD3 antibody and / or anti-CD28 antibody). 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 the immune cells with a Cbl-b inhibitor or a composition thereof, thereby generating modified immune cells. In some embodiments, the modified 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 immune cells are of the same cell type as the modified immune cells. For example, the immune cells can be inactivated T cells, and the modified immune cells can be activated T cells. In some embodiments, the immune cells are of a different cell type from the modified immune cells. For example, the immune cells can be hematopoietic stem cells, and the modified immune cells can be NK cells differentiated from the hematopoietic stem cells. In some embodiments of methods for generating modified immune cells, the method further comprises recovering the modified immune cells. In some embodiments, the cell population comprising the immune cells, the immune cells, or the modified immune cells are all from an individual (e.g., a human). In some embodiments, the immune cells or the modified immune cells are human immune cells or human modified immune cells, respectively.

[0314] The present invention further provides modified immune cells generated by any of the methods described in the present invention, for example, culturing a cell population comprising immune cells in the presence of an effective amount of a Cbl-b inhibitor to regulate the activity of the immune cells, thereby generating the modified immune cells.

[0315] In some embodiments, the Cbl-b inhibitor provided by the present invention is cell membrane permeable. Thus, in some embodiments, the modified immune cells provided by the present invention may comprise the Cbl-b inhibitor described in the present invention, for example, the Cbl-b inhibitor described in the present invention is comprised in the cytoplasm of the modified immune cells.

[0316] In some cases, the present invention provides isolated modified immune cells, wherein the modified immune cells have been contacted with or are contacted with the Cbl-b inhibitor or its composition as described in the present invention. 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.

[0317] In some embodiments of the isolated modified immune cells, the modified immune cells are T cells, and the T cells exhibit increased T cell activation, increased T cell proliferation, decreased T cell exhaustion, and / or reduced T cell tolerance. In some embodiments, increased T cell activation includes an increase in the production of one or more cytokines from T cells or surrounding 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 one or more cytokines are one or more selected from the group consisting of: 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, eosinophil chemotactic factor (Eotaxin), GROα, RANTES, MIP-1α, MIP-1β, MIP-2, MCP-1, and MCP-3. In some embodiments, increased T cell activation includes an increase in 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 or are contacted with an anti-CD3 antibody. In some embodiments, the T cells have been contacted with or are contacted with a combination of an anti-CD3 antibody and an anti-CD28 antibody.

[0318] In some embodiments of the isolated modified immune cells, the modified immune cells are NK cells, and the NK cells exhibit increased NK cell activation. In some embodiments, the increased NK cell activation comprises increased secretion of one or more cytokines (e.g., IFN-γ, TNFα, and / or MIP-1β). In some embodiments, the increased NK cell activation comprises increased cell surface expression of one or more NK cell activation markers (e.g., CD69 and / or CD107a).

[0319] In some embodiments of the isolated modified immune cells, the modified immune cells are B cells, and the B cells exhibit increased B cell activation. In some embodiments, the increased B cell activation comprises increased cell surface expression of one or more B cell activation markers (e.g., CD69, CD86, and / or HLA-DR).

[0320] In some of any of the embodiments of the methods or modified immune cells provided by the present invention, the immune cells or modified immune cells are mammalian cells (e.g., human cells). In some embodiments, the immune cells or modified immune cells are human cells.

[0321] In some cases, the incubation is carried out according to techniques such as those described in U.S. Patent No. 6,040,177; Klebanoff et al., J Immunol Ther., 35:651-660, 2012; Terakura et al., Blood, 119:72-82, 2012; and Wang et al., J Immunol Ther., 35:689-701, 2012.

[0322] The immune cells to be modified or the 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, the 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 activating cytoplasmic signaling domain. In some embodiments, the CAR comprises, from its N-terminus to its C-terminus: an extracellular ligand-binding domain, a transmembrane domain, and an activating cytoplasmic signaling domain. The immune cells can be engineered to express the 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 further embodiments, the modified T cells comprise a recombinant chimeric receptor, such as a CAR. Methods for generating immune cells expressing a recombinant chimeric receptor are well known in the art, for example by introducing a nucleic acid encoding the recombinant chimeric receptor (e.g., a CAR) into an immune cell (e.g., a T cell) via a vector (e.g., a viral vector). See, for example, International Patent Application No. WO2017 / 096329 and U.S. Publication No. US2017 / 0204372.

[0323] In particular, the present invention provides a method for generating an expanded population of lymphocytes, the method comprising: (a) obtaining a biological sample comprising lymphocytes from an individual with 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 the lymphocytes in a cell culture medium comprising at least one T cell growth factor to generate an expanded population of lymphocytes. In certain embodiments, the lymphocytes are tumor infiltrating lymphocytes (TIL). In certain embodiments, the lymphocytes are peripheral blood mononuclear cells (PBMC). In certain 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 population of TIL generated by the above method and a physiologically acceptable buffer.

[0324] In some embodiments, methods for isolating and processing immune cells to be modified or that have been modified (i.e., modified immune cells) include the 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 an immune cell). A variety of cryopreservation solutions and parameters known in the art can be used.

[0325] B. Adoptive cell therapy

[0326] The modified immune cells, such as an expanded lymphocyte population or a composition thereof generated by the methods described herein, can be used as therapeutic agents in methods for treating an individual in need (e.g., an individual suffering from cancer). Such treatment methods include adoptive cell therapy. In some embodiments, the treatment method includes isolating cells from an individual, preparing, processing, 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, processing, culturing, and / or engineering them as described herein, and reintroducing them into a different individual before or after cryopreservation.

[0327] Thus, in some cases, the present invention provides methods for modulating an immune response in an individual, the method comprising administering an effective amount of the modified immune cells or a composition thereof described herein to an individual in need (e.g., an individual suffering from T cell dysfunction). In some embodiments, the individual has cancer. In some embodiments, the present invention provides a method for treating a cancer responsive to inhibition of Cbl-b activity, the method comprising administering an effective amount of the modified immune cells or a composition thereof described herein to an individual suffering from the cancer responsive to inhibition of Cbl-b activity. In some embodiments, the present invention provides a method for inhibiting abnormal cell proliferation, the method comprising administering an effective amount of the modified immune cells or a composition thereof described herein to an individual in need. As used herein, the term "abnormal cell proliferation" includes hyperplasia or cancer cell proliferation. Cancer cells can be derived from hematological cancers, such as lymphoma, leukemia, or myeloma. In other embodiments, cancer cells can be derived from non-hematological cancers, such as sarcoma, carcinoma, or melanoma.

[0328] In certain embodiments, a composition is administered to an individual in need of treatment (e.g., an individual suffering from cancer or T cell dysfunction), the composition comprising from about 1 million to about 100 billion cells, such as from 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), such as from 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 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, from about 100 million 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 a modified immune cell provided by the present invention having any value between these ranges.

[0329] The modified immune cells and their compositions 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. Formulations 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 a sterile liquid formulation, such as an isotonic aqueous solution, suspension, emulsion, dispersion, or viscous composition, which in some cases can be buffered to a selected pH value. The viscous composition can be formulated within a suitable viscosity range to provide a longer contact time with a particular tissue. The liquid or viscous composition can contain a carrier, which can be a solvent or dispersion medium comprising, for example, water, saline, phosphate buffered saline, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. A sterile injectable solution can be prepared by incorporating the modified immune cells into a solvent, such as by mixing with a suitable carrier, diluent, or excipient (e.g., sterile water, saline, dextrose, dextran, or the like).

[0330] In some embodiments, the modified immune cells are co-administered with one or more additional therapeutic agents or in combination with 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 represented by formula (I), (I-a), (I-b), (I-A)-(I-J), (II-A)-(II-H), (III-A)-(III-H), or (IV-A)-(IV-H), or any variant thereof. Thus, in some embodiments, the treatment regimen comprises both adoptive cell therapy and chemotherapy.

[0331] After administering the modified immune cells to an individual (e.g., a human), the biological activity of the population of modified immune cells can be determined by methods known in the art. Parameters to be evaluated include specific binding of the modified immune cells or other immune cells to an antigen in vivo (e.g., by imaging) or ex vivo (e.g., by ELISA or flow cytometry). In some embodiments, cytotoxicity assays can be used to determine the ability of the 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 biological activity of the modified immune cells can also be determined by measuring the expression and / or secretion of certain cytokines (e.g., IL-2 and IFNγ).

[0332] C. Administration of Cbl-b Inhibitors

[0333] In some cases, a Cbl-b inhibitor or a composition thereof can be directly administered to an individual to modulate the immune response, treat a disease or disorder (e.g., cancer and / or abnormal cell proliferation), and / or inhibit the Cbl-b activity of the individual. The Cbl-b inhibitor can be a compound shown in Table 1, a tautomer thereof, or a pharmaceutically acceptable salt of any of the foregoing substances.

[0334] In some embodiments, the present invention provides a method of modulating an immune response, the method comprising administering to an individual an effective amount of a Cbl-b inhibitor or a composition thereof provided by the present invention to modulate the immune response of the individual. In some embodiments, the individual has cancer, such as a hematological cancer or a non-hematological cancer as described in the present invention.

[0335] In some embodiments, the present invention provides a method for treating cancer responsive to inhibition of Cbl-b activity, the method comprising administering to an individual an effective amount of a Cbl-b inhibitor or a composition thereof provided by the present invention to treat cancer responsive to inhibition of Cbl-b activity. In some embodiments, the cancer is a hematological cancer or a non-hematological cancer, such as the cancers described in the present invention.

[0336] In some embodiments, the present invention provides a method for inhibiting abnormal cell proliferation (e.g., hyperplasia), the method comprising administering to an individual an effective amount of a Cbl-b inhibitor or a composition thereof provided by the present invention to inhibit abnormal cell proliferation in the individual.

[0337] In some embodiments, the present invention provides a method for inhibiting Cbl-b activity, the method comprising administering to an individual an effective amount of a Cbl-b inhibitor or a composition thereof provided by the present invention to inhibit Cbl-b activity in the individual.

[0338] In some embodiments, such as in modulating the immune response of an individual in need (e.g., an individual suffering from T cell dysfunction), treating a disease or disorder of an individual (e.g., cancer and / or abnormal cell proliferation of an individual), and / or inhibiting Cbl-b activity of an individual, the appropriate dose of the active agent will depend on the condition to be treated, the type of disease or disorder (as defined above), the severity and course of the condition, disease or disorder, whether the agent is administered for prophylactic or therapeutic purposes, previous treatment, the clinical history of the subject and the response to the Cbl-b inhibitor, and the judgment of the attending physician.

[0339] The Cbl-b inhibitor or a composition thereof is appropriately administered to an individual once or in a series of treatments. In some embodiments, the treatment includes multiple administrations of the Cbl-b inhibitor or a composition thereof, wherein the interval between administrations can vary. For example, the interval between the first administration and the second administration 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 an individual at a fixed dose based on the body weight of the individual (e.g., mg / kg).

[0340] In some cases of the present invention, the cancer is a hematological cancer. For example, the hematological cancer can be lymphoma, leukemia, or myeloma. In other cases of the present invention, the cancer is a non-hematological cancer. In particular, the non-hematological cancer can be carcinoma, sarcoma, or melanoma.

[0341] In some embodiments, the Cbl-b inhibitor is co-administered with one or more additional therapeutic agents or in combination with another therapeutic intervention, either simultaneously or sequentially in any order. For example, in some treatment 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 represented by formula (I), (I-a), (I-b), (I-A)-(I-J), (II-A)-(II-H), (III-A)-(III-H), or (IV-A)-(IV-H), or any variant thereof. Thus, in some embodiments, the treatment regimen comprises both adoptive cell therapy and chemotherapy.

[0342] In some embodiments, the efficacy of administering a Cbl-b inhibitor in the methods of the present invention (e.g., methods of modulating an individual's immune response) can be evaluated by determining the biological activity of immune cells present in a sample (e.g., a blood sample) isolated from the treated individual. For example, the therapeutic efficacy can be evaluated by determining the ability of immune cells isolated from an individual to destroy target cells after treatment with a Cbl-b inhibitor in a cytotoxicity assay. In some embodiments, the biological activity of immune cells present in a sample (e.g., a blood sample) can be determined by measuring the expression and / or secretion of certain cytokines (e.g., IL-2 and IFNγ).

[0343] The present invention provides methods for treating cancer, comprising: administering to an individual suffering from cancer an effective amount of a Cbl-b inhibitor, and administering to the individual an effective amount of an additional therapeutic agent. The present invention also provides methods for treating an individual suffering from cancer, comprising: administering to the individual an effective amount of a Cbl-b inhibitor; and administering to the individual an effective amount of an additional therapeutic agent. In addition, the present invention provides methods for enhancing an anti-cancer immune response, comprising: administering to an individual suffering from cancer an effective amount of a Cbl-b inhibitor, and administering to the individual an effective amount of an additional therapeutic agent. The present invention also provides methods for treating cancer, comprising: administering to an individual suffering from cancer an effective amount of a Cbl-b inhibitor, wherein the individual has received or is receiving an effective amount of an additional therapeutic agent.

[0344] In some embodiments of the methods described in the foregoing paragraphs, the Cbl-b inhibitor and the additional therapeutic agent are administered sequentially in either order. As used herein, the terms "sequentially," "consecutively," and "in sequence" mean that the Cbl-b inhibitor is administered after the additional therapeutic agent, or the additional therapeutic agent is administered after the Cbl-b inhibitor. For example, sequential administration can include administering the Cbl-b inhibitor during an induction period (initial treatment) in the absence of the additional therapeutic agent, followed by a post-induction treatment period that includes administering the additional therapeutic agent. The method can further include a maintenance phase that includes administering the Cbl-b inhibitor or the additional therapeutic agent. Alternatively, sequential administration can include administering the additional therapeutic agent during an induction period (initial treatment) in the absence of the Cbl-b inhibitor, followed by a post-induction treatment period that includes administering the Cbl-b inhibitor. The method can further include a maintenance phase that includes administering the Cbl-b inhibitor or the additional therapeutic agent.

[0345] In some embodiments of the combination therapy methods, the Cbl-b inhibitor and the additional therapeutic agent are administered simultaneously. As used herein, the terms "simultaneously," "concurrently," and "in parallel" mean that the Cbl-b inhibitor and the additional therapeutic agent are administered during the same physician 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 period, treatment period, and maintenance period. However, simultaneous administration does not require that the Cbl-b inhibitor and the additional therapeutic agent be present in a single formulation or pharmaceutical composition together, or that the Cbl-b inhibitor and the additional therapeutic agent be administered precisely at the same time.

[0346] 1. Combination therapy comprising a Cbl-b inhibitor and an immune checkpoint inhibitor

[0347] In some embodiments of the combination therapy methods of the present invention, the additional therapeutic agent comprises an immune checkpoint inhibitor. The term "immune checkpoint" refers to signaling pathways that prevent the activation of immune cells, while the term "immune checkpoint inhibitor" refers to compounds that block immune checkpoints to relieve the brake 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 selected from the group consisting of PD-1 (CD279), PD-L1 (CD274), and CTLA-4 (CD152).

[0348] PD-1 refers to programmed cell death protein 1 (PD-1). PD-1 antagonists suitable for the treatment methods, drugs, and uses of the present invention include any chemical compound or biomolecule 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 ligand include: for PD-1, there are CD279, PDCD1, PD1, and SLEB2; and for programmed cell death 1 ligand 1 (PD-L1), there are CD274, PDCD1L1, PDL1, B7H1, B7-4, and B7-H. 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 number NP_054862.

[0349] CTLA-4 refers to cytotoxic T lymphocyte-associated protein 4. CTLA-4 antagonists suitable for the treatment methods, drugs, and uses of the present invention include any chemical compound or biomolecule that blocks 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 of CTLA-4 include: CD152, CTLA4, ALPS5, CELIAC3, GRD4, GSE, and IDDM12. In some embodiments of treating human subjects, the CTLA-4 antagonist blocks 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.

[0350] LAG3 refers to lymphocyte activation gene 3 protein. LAG3 antagonists suitable for the treatment methods, drugs, 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 of treating human subjects, the LAG3 antagonist blocks the binding of human LAG3 to human ligands. The amino acid sequence of the mature form of human LAG3 is shown as residues 23-525 in NCBI locus number NP_002277.

[0351] PVR refers to poliovirus receptor. PVR antagonists suitable for the treatment methods, drugs, 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 of PVR include CD155, PVS, HVED, NECL5, nectin-like protein 5, and TAGE4. In some embodiments of treating human subjects, the PVR antagonist blocks the binding of human PVR to human TIGIT. There are multiple isoforms of human PVR. 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 δ isoform of human PVR is shown in NCBI locus number NP_001129242.

[0352] PVRL2 refers to poliovirus receptor-related 2. PVRL2 antagonists suitable for the treatment methods, drugs, and uses of the present invention include any chemical compound or biomolecule 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 of PVRL2 include: CD112, NECTIN2, HVEB, herpesvirus entry mediator B, PRR2, and PVRR2. In some embodiments of treating human subjects, the PVRL2 antagonist blocks the binding of human PVRL2 to human TIGIT. The amino acid sequence of the α isoform of human PVRL2 is shown in NCBI locus number NP_002847. The amino acid sequence of the δ isoform of human PVRL2 is shown in NCBI locus number NP_001036189.

[0353] PVRL3 refers to poliovirus receptor-related 3. PVRL3 antagonists suitable for the treatment methods, drugs, and uses of the present invention include any chemical compound or biomolecule 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 of PVRL3 include: CD113, NECTIN3, PRR3, and PVRR3. In some embodiments of treating human subjects, the PVRL3 antagonist blocks the binding of human PVRL3 to human TIGIT. The amino acid sequence of isoform 1 of human PVRL3 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.

[0354] TIGIT refers to T cell immunoreceptor with Ig and ITIM domains protein. TIGIT antagonists suitable for the treatment methods, drugs, and uses of the present invention include any chemical compound or biomolecule 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 of TIGIT include: VSIG9, V-set and immunoglobulin domain-containing protein 9, VSTM3, V-set and transmembrane domain-containing protein 3, and Washington University cell adhesion molecule (WUCAM). In some embodiments of treating human subjects, the TIGIT antagonist blocks the binding of human TIGIT to human ligands. The amino acid sequence of the mature form of human TIGIT is shown as residues 22 - 244 in NCBI locus number: NP_776160.

[0355] TIM3 refers to T-cell immunoglobulin and mucin-domain containing-3 protein. TIM3 antagonists suitable for the treatment methods, drugs 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 ligands (galectin-9, phosphatidylserine) expressed on antigen-presenting cells. Alternative names or synonyms of TIM3 include: CD366, HAVCR2, hepatitis A virus cellular receptor 2, KIM3, and SPTCL. In some embodiments of treating human subjects, the TIM3 antagonist blocks the binding of human TIM3 to human ligands. The amino acid sequence of the mature form of human TIM3 is shown as residues 22-301 in NCBI locus number NP_116171.

[0356] VISTA refers to V-domain Ig suppressor of T cell activation. VISTA antagonists suitable for the treatment methods, drugs 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 of VISTA include: VSIR, V-set immunoregulatory receptor, PD-1H, B7H5, GI24, PP2135, SISP1, and Dies1. In some embodiments of treating human subjects, the VISTA antagonist blocks the binding of human VISTA to human ligands. The amino acid sequence of the mature form of human VISTA is shown as residues 33-311 in NCBI locus number: NP_071436.

[0357] The immune checkpoint inhibitor can be a biomolecule. For example, the immune checkpoint inhibitor can comprise an antibody or an antigen-binding fragment thereof. The antibody or fragment can be a monoclonal antibody (mAb), a human antibody, a humanized antibody, or a chimeric antibody, and can include a human constant region. In some embodiments, the human constant region is selected from the group consisting of IgGl, IgG2, IgG3, and IgG4 constant regions, and in certain embodiments, the human constant region is the IgGl 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.

[0358] 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 their biosimilars. In one embodiment, the anti-PD-1 antibody is pembrolizumab (MK-3475, sold by Merck & Co. as ). In one embodiment, the anti-PD-1 antibody is nivolumab (BMS-936558 or MDX-1106, sold 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 a variant of pembrolizumab, nivolumab, or cemiplimab.

[0359] 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 their biosimilars. In one embodiment, the anti-PD-L1 antibody is atezolizumab (sold by Genentech, Inc. as ). In one embodiment, the anti-PD-L1 antibody is avelumab (sold by EMD Serono, Inc. and Pfizer, Inc. as ). In one embodiment, the anti-PD-L1 antibody is durvalumab (MEDI4736, sold by AstraZeneca as ). In some embodiments, the immune checkpoint inhibitor is a variant of atezolizumab, avelumab, or durvalumab.

[0360] 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 their biosimilars. In one embodiment, the anti-CTLA4 antibody is ipilimumab (MDX-010 or BMS-734016, sold by Bristol-Myers Squibb under ). In one embodiment, the anti-CTLA4 antibody is tremelimumab (ticilimumab, CP-675,206, developed by AstraZeneca). In some embodiments, the immune checkpoint inhibitor is a variant of ipilimumab or tremelimumab.

[0361] In some embodiments, the monoclonal antibody is a "variant" antibody that comprises heavy and light chain sequences that are identical to the heavy and light chain sequences in a "reference" antibody, except for having 3, 2, or 1 conservative amino acid substitutions at positions located outside the light chain CDRs and / or 6, 5, 4, 3, 2, or 1 conservative amino acid substitutions at positions located outside the heavy chain CDRs (e.g., the variant positions are all located in the framework region or the constant region). In other words, the reference antibody and the variant antibody both comprise the same CDR sequences, but differ from each other by having no more than 3 or 6 conservative amino acid substitutions at 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 the following characteristics: the binding affinity for the inhibitory checkpoint molecule and the ability to block the binding of the inhibitory checkpoint molecule to its ligand.

[0362] In other embodiments, the immune checkpoint inhibitor may comprise an immunoadhesin that comprises an inhibitory checkpoint molecule binding domain fused to a constant region (e.g., the Fc region of an immunoglobulin molecule).

[0363] As used herein, the term "biosimilar" or "biological equivalent" refers to a biological product that is similar to a reference product approved by the US Food and Drug Administration (FDA) but has no clinically meaningful differences in terms of safety and efficacy. For example, there may be differences in the clinically inactive ingredients between a biosimilar product and the reference product (e.g., differences in formulation excipients, minor differences in glycosylation, etc.). Clinically meaningful characteristics can be evaluated through pharmacokinetic and pharmacodynamic studies. In some embodiments, the biosimilar product is a product determined by the FDA to be interchangeable.

[0364] 2. Combination Therapy Comprising a Cbl-b Inhibitor and an Antitumor Agent

[0365] In some embodiments of the combination therapy methods of the present invention, the additional therapeutic agent comprises an antitumor agent. As used herein, the terms "antitumor 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 antitumor 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 antitumor agents (ATC code L01X). In some embodiments, the antitumor agent is a small molecule drug (e.g., a cancer chemotherapeutic agent), rather than a biomolecule.

[0366] Cytotoxic antibiotics are antitumor agents suitable for the treatment methods, drugs, and uses of the present invention. In some embodiments, the cytotoxic antibiotics are selected from the group consisting of: ixabepilone, mitomycin, plicamycin, bleomycin, pixantrone, amrubicin, valrubicin, pirarubicin, mitoxantrone, idarubicin, zorubicin, aclarubicin, epirubicin, daunorubicin, doxorubicin, and dactinomycin.

[0367] Plant alkaloids are antitumor agents suitable for the treatment methods, drugs, 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.

[0368] Antimetabolites are anti-tumor agents applicable to the treatment methods, drugs and uses of the present invention. In some embodiments, the antimetabolite is a pyrimidine analogue, a purine analogue or a folic acid analogue. 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.

[0369] Alkylating agents are anti-tumor agents suitable for the treatment methods, drugs 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, ranimustine, nimustine, fotemustine, streptozocin, semustine, lomustine, carmustine, carboquone, triaziquone, thiotepa, mannosulfan, treosulfan, busulfan, bendamustine, prednimustine, trofosfamide, ifosfamide, mechlorethamine, melphalan, chlorambucil, and cyclophosphamide.

[0370] In other embodiments, the anti-tumor agent comprises other anti-tumor agents 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).

[0371] Platinum compounds are suitable anti-tumor agents that can be used in the treatment methods, drugs 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.

[0372] 3. Combination therapy comprising a Cbl-b inhibitor and radiotherapy

[0373] The present invention provides methods for treating cancer, comprising: administering to an individual suffering from cancer an effective amount of a Cbl-b inhibitor, and administering to the individual an effective amount of radiotherapy. The present invention also provides methods for treating an individual suffering from cancer, comprising: administering to the individual an effective amount of Cbl-b, and administering to the individual an effective amount of radiotherapy. In addition, the present invention provides methods for increasing an anti-cancer immune response, comprising: administering to an individual suffering from cancer an effective amount of a Cbl-b inhibitor, and administering to the individual an effective amount of radiotherapy. The present invention also provides methods for treating cancer, comprising: administering to an individual suffering from cancer an effective amount of a Cbl-b inhibitor, wherein the individual has received or is receiving an effective amount of radiotherapy.

[0374] In some embodiments, the radiotherapy is external beam radiotherapy. In other embodiments, the radiotherapy is brachytherapy. In some embodiments, the radiotherapy is ablative radiotherapy.

[0375] In some embodiments, the combination therapy regimens of the present invention comprise administering a Cbl-b inhibitor, radiotherapy, and one or both of an immune checkpoint inhibitor and an anti-tumor agent.

[0376] The present invention provides methods 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, and 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. The present invention also provides methods 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 an oncolytic virus. The present invention also provides a medicament comprising a Cbl-b inhibitor for use in combination with an oncolytic virus to treat cancer, and a medicament 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 medicament for treating cancer in an individual 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 medicament for treating cancer. In some embodiments, the Cbl-b inhibitor is a "small molecule".

[0377] In some embodiments of the treatment methods, drugs, and uses of the present invention, the cancer is a hematological cancer, such as lymphoma, leukemia, or myeloma. In other embodiments of the treatment methods, drugs, and uses of the present invention, the cancer is a non-hematological cancer, such as sarcoma, carcinoma, or melanoma.

[0378] 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, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, follicular small cell lymphoma or follicular large cell lymphoma, malignant lymphoproliferative disease, 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 “preleukemia”, which are all various hematological diseases united by ineffective production (or dysplasia) of myeloid blood cells.

[0379] 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.

[0380] In some cases, the effectiveness of administering an activation threshold lowering agent or a co-stimulation requirement lowering agent, such as a Cbl-b inhibitor, in treating cancer is determined by evaluating clinical outcomes (e.g., reduction in tumor size or number of tumors, and / or survival rate). In some embodiments, "treating cancer" includes 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). The response evaluation criteria for determining objective anti-tumor response according to RECIST 1.1 include: complete response (CR); partial response (PR); progressive disease (PD); and stable disease (SD).

[0381] The present invention also provides a method for treating cancer, which comprises administering to an individual suffering from cancer a combination therapy, said combination therapy comprising an agent (activation threshold reducing agent) effective to reduce 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 medicament comprising an activation threshold reducing agent for use in combination with a cancer vaccine for treating cancer, and a medicament for treating cancer comprising both an activation threshold reducing agent and a cancer vaccine. The present invention further provides the use of an activation threshold reducing agent in the preparation of a medicament for treating an individual's cancer when administered in combination with a cancer vaccine. The present invention further provides the use of an activation threshold reducing agent and a cancer vaccine in the preparation of a medicament for treating cancer. The present invention also provides a method for treating cancer, which comprises administering to an individual suffering from cancer a combination therapy, said combination therapy comprising an agent (activation threshold reducing agent) effective to reduce 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 medicament comprising an activation threshold reducing agent for use in combination with an oncolytic virus for treating cancer, and a medicament for treating cancer comprising both an activation threshold reducing agent and an oncolytic virus. The present invention further provides the use of an activation threshold reducing agent in the preparation of a medicament for treating an individual's cancer when administered in combination with an oncolytic virus. The present invention further provides the use of an activation threshold reducing agent and an oncolytic virus in the preparation of a medicament for treating cancer. In some embodiments, the agent (activation threshold reducing agent) that reduces the activation threshold is an agent that reduces the co-stimulation requirement (co-stimulation requirement reducing agent) of immune cells (e.g., T cells, B cells, and / or NK cells). In some embodiments, the agent (activation threshold reducing agent) that reduces the activation threshold is an agent that promotes tumor immune surveillance. In some embodiments, the agent (activation threshold reducing agent) that reduces the activation threshold is a Cbl-b inhibitor. In some embodiments, the agent that reduces the co-stimulation requirement is a Cbl-b inhibitor. In some embodiments, the agent that promotes tumor immune surveillance is a Cbl-b inhibitor.

[0382] In some embodiments, an activation threshold reducing agent, such as a Cbl-b inhibitor, is capable of increasing T cell activation and / or T cell proliferation. In some embodiments, an activation threshold reducing agent, such as a Cbl-b inhibitor, is capable of reducing T cell exhaustion, T cell tolerance, and / or T cell anergy.

[0383] In some embodiments, an activation threshold lowering agent, such as a Cbl-b inhibitor, is capable of increasing the production of one or more cytokines in the activated T cell microenvironment by T cells or surrounding immune cells (e.g., myeloid cells). 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. The increased cytokine expression can be measured by ELISA.

[0384] In some embodiments, an activation threshold lowering agent, such as a Cbl-b inhibitor, is capable of 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 T cell activation marker is one or more of the following: CD25, CD69, and CTLA4. The increased expression of cell surface markers can be measured by FACS.

[0385] 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 12 provided in the present invention. 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 13.

[0386] In some embodiments, an activation threshold lowering agent, such as a Cbl-b inhibitor, is capable of increasing 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. In some embodiments, the one or more B cell activation markers include, but are not limited to: CD69, CD86, and major histocompatibility complex class II (e.g., HLA-DR). In some embodiments, the B cell activation marker is CD69. The increased expression of the cell surface marker can be measured by FACS. In some embodiments, the increased B cell activation comprises an increase in protein activation in signaling pathways (e.g., those mediated by ERK, JNK, and Syk). The increased protein activation can be detected by measuring the phosphorylation level on the protein using reagents (e.g., anti-phosphorylation antibodies available in the art).

[0387] In some embodiments, an activation threshold lowering agent, such as a Cbl-b inhibitor, is capable of increasing 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 an increase in 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 the cell surface marker can be measured by FACS. In some embodiments, the increased NK cell activation comprises an increase in the killing of target cells (e.g., tumor cells, including primary tumor cells, and cell line-derived tumor cells, such as the K562 cell line).

[0388] Methods for experimentally determining increased B cell activation and NK cell activation are well known in the art.

[0389] The activity regulation of immune cells (e.g., T cells, B cells, or NK cells) can be determined by measuring the baseline value of a 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 in contact with a Cbl-b inhibitor, can be measured before contacting or administering the Cbl-b inhibitor to determine the baseline value. Then a reference value of T cell activation is obtained after contacting or administering 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 administering the Cbl-b inhibitor or its composition. For example, in some embodiments, the activation of immune cells (e.g., T cells) in the sample increases by at least 0.1-fold compared to the baseline value, where the baseline value is obtained before contacting the immune cells (e.g., T cells) with the Cbl-b inhibitor or its composition. In some embodiments, the activation of immune cells (e.g., T cells) increases by at least about 0.1-fold, about 0.2-fold, about 0.3-fold, about 0.4-fold, about 0.5-fold, about 0.6-fold, about 0.7-fold, about 0.8-fold, about 0.9-fold, about 1-fold, about 2-fold, about 4-fold, about 6-fold, about 8-fold, about 10-fold, about 20-fold, about 30-fold, but not more than about 50-fold compared to the baseline value. The activation of immune cells can be evaluated by measuring an increase in the secretion of activation biomarkers such as cytokines, an increase in the cell surface expression of activation markers (e.g., cell surface markers), or an increase in the phosphorylation of proteins in downstream signaling pathways. The fold change relative to the baseline value indicating immune cell activation can be determined for the parameter being tested and the conditions under which the immune cells are treated. For example, to measure T cell activation, a baseline value can be obtained from T cells stimulated with a combination of anti-CD3 antibody and anti-CD28 antibody, where the cells are not incubated with the Cbl-b inhibitor. Then a reference value is obtained from T cells stimulated with a combination of anti-CD3 antibody and anti-CD28 antibody, where the T cells have been in contact with or contacted the Cbl-b inhibitor. Then a positive response of immune cell activation can be determined by the obtained reference value. Similar reference value measurements can be obtained and compared with the baseline values used to evaluate 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.

[0390] As used herein, the term "baseline" or "baseline value" may refer to a measured value or characterization before administration of a therapeutic agent disclosed herein (e.g., a composition comprising a Cbl-b inhibitor as described herein) or at the start of administration of said therapeutic agent. The baseline value may 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 reduced T cell tolerance). As used herein, the term "reference" or "reference value" may refer to a measured value or characterization after administration of a therapeutic agent disclosed herein (e.g., a composition comprising a Cbl-b inhibitor as described herein). The reference value may be measured one or more times during an experimental time course, dosing regimen, or treatment cycle or at the completion of an experimental time course, dosing regimen, or treatment cycle. A "reference value" may 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, a mean value, or a value compared to a baseline value. Similarly, a "baseline value" may 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, a mean value, or a value compared to a reference value. The reference value and / or baseline value may 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 set of samples (e.g., samples obtained from a set of two, three, four, five, or more individuals).

[0391] In some embodiments, the 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 antibody and anti-CD28 antibody in the presence of a Cbl-b inhibitor is at least 2.5-fold the baseline value of cytokine secretion (e.g., IL-2 secretion) obtained from T cells stimulated with a combination of anti-CD3 antibody and anti-CD28 antibody in the absence of a Cbl-b inhibitor. In some embodiments, the positive response to T cell activation as measured by surface marker expression (e.g., CD25 surface marker staining) of T cells stimulated with a combination of anti-CD3 antibody and anti-CD28 antibody in the presence of a Cbl-b inhibitor is at least 1.3-fold the baseline value of surface marker expression (e.g., CD25 surface marker staining) obtained from T cells stimulated with a combination of anti-CD3 antibody and anti-CD28 antibody in the absence of a Cbl-b inhibitor. In some embodiments, the baseline value can be obtained from T cells stimulated with anti-CD3 antibody alone, wherein the cells are not incubated with a Cbl-b inhibitor. In some embodiments, the positive response to T cell activation as measured by cytokine secretion (e.g., IL-2 secretion) of T cells stimulated with anti-CD3 antibody alone in the presence of a Cbl-b inhibitor is at least 0.1-fold the baseline value of cytokine secretion (e.g., IL-2 secretion) obtained from T cells stimulated with anti-CD3 antibody alone in the absence of a Cbl-b inhibitor. In some embodiments, the positive response to T cell activation as measured by surface marker expression (e.g., CD25 surface marker staining) of T cells stimulated with anti-CD3 antibody alone in the presence of a Cbl-b inhibitor is at least 0.6-fold the baseline value of surface marker expression (e.g., CD25 surface marker staining) obtained from T cells stimulated with anti-CD3 antibody alone in the absence of a Cbl-b inhibitor.

[0392] The present invention provides methods for treating cancer, comprising: administering to an individual suffering from cancer an effective amount of a Cbl-b inhibitor, and administering to said individual an effective amount of a cancer vaccine. The present invention also provides methods for treating an individual suffering from cancer, comprising: administering to said individual an effective amount of a Cbl-b inhibitor; and administering to said individual an effective amount of a cancer vaccine. In addition, the present invention provides methods for enhancing an anti-cancer immune response, comprising: administering to an individual suffering from cancer an effective amount of a Cbl-b inhibitor, and administering to said individual an effective amount of a cancer vaccine. The present invention further provides methods for treating cancer, comprising: administering to an individual suffering from cancer an effective amount of a Cbl-b inhibitor, wherein said individual has received or is receiving an effective amount of a cancer vaccine. In addition, the present invention provides methods for treating cancer, comprising: administering to an individual suffering from cancer an effective amount of a Cbl-b inhibitor, and administering to said individual an effective amount of an oncolytic virus. The present invention also provides methods for treating an individual suffering from cancer, comprising: administering to said individual an effective amount of a Cbl-b inhibitor; and administering to said individual an effective amount of an oncolytic virus. In addition, the present invention provides methods for enhancing an anti-cancer immune response, comprising: administering to an individual suffering from cancer an effective amount of a Cbl-b inhibitor, and administering to said individual an effective amount of an oncolytic virus. The present invention also provides methods for treating cancer, comprising: administering to an individual suffering from cancer an effective amount of a Cbl-b inhibitor, wherein said individual has received or is receiving an effective amount of an oncolytic virus.

[0393] 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. In certain embodiments, as used herein, the terms "sequentially," "consecutively," and "in sequence" mean that the Cbl-b inhibitor is administered after the cancer vaccine, or the cancer vaccine is administered after the Cbl-b inhibitor. For example, sequential administration can include administering the Cbl-b inhibitor during an induction period (initial treatment) in the absence of the cancer vaccine, followed by a post-induction treatment period that includes administering both the cancer vaccine and the Cbl-b inhibitor. The method can further include a maintenance phase that includes administering the Cbl-b inhibitor or an additional dose of the cancer vaccine. Alternatively, sequential administration can include administering the cancer vaccine during an induction period (initial treatment) in the absence of the Cbl-b inhibitor, followed by a post-induction treatment period that includes administering the Cbl-b inhibitor. The method can further include a maintenance phase that includes administering the Cbl-b inhibitor or an additional dose of the cancer vaccine. In some embodiments of the methods described in the preceding paragraphs, the Cbl-b inhibitor and the oncolytic virus are administered sequentially in either order. In certain embodiments, as used herein, the terms "sequentially," "consecutively," and "in sequence" mean that the Cbl-b inhibitor is administered after the oncolytic virus, or the oncolytic virus is administered after the Cbl-b inhibitor. For example, sequential administration can include administering the Cbl-b inhibitor during an induction period (initial treatment) in the absence of the oncolytic virus, followed by a post-induction treatment period that includes administering both the oncolytic virus and the Cbl-b inhibitor. The method can further include a maintenance phase that includes administering the Cbl-b inhibitor or an additional dose of the oncolytic virus. Alternatively, sequential administration can include administering the oncolytic virus during an induction period (initial treatment) in the absence of the Cbl-b inhibitor, followed by a post-induction treatment period that includes administering the Cbl-b inhibitor. The method can further include a maintenance phase that includes administering the Cbl-b inhibitor or an additional dose of the oncolytic virus.

[0394] In some embodiments of the combination therapy methods, the Cbl-b inhibitor and the cancer vaccine are administered simultaneously. In certain embodiments, as used herein, the terms "simultaneously", "concurrently", and "in parallel" mean that the Cbl-b inhibitor and the cancer vaccine are administered during the same doctor 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, treatment, and maintenance phases. 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 precisely at the same time. In some embodiments of the combination therapy methods, the Cbl-b inhibitor and the oncolytic virus are administered simultaneously. In certain embodiments, as used herein, the terms "simultaneously", "concurrently", and "in parallel" mean that the Cbl-b inhibitor and the oncolytic virus are administered during the same doctor 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, treatment, and maintenance phases. However, simultaneous administration does not require that the Cbl-b inhibitor and the oncolytic virus be present together in a single formulation or pharmaceutical composition, or that the Cbl-b inhibitor and the oncolytic virus be administered precisely at the same time.

[0395] In some cases, the treatment includes multiple administrations of the Cbl-b inhibitor or its composition, where the intervals between administrations can vary. In some embodiments, the Cbl-b inhibitor is administered to an individual at a fixed dose (e.g., mg / adult or mg / child). In some embodiments, the Cbl-b inhibitor is administered to an individual at a fixed dose based on the individual's body weight (e.g., mg / kg).

[0396] In some embodiments, the effectiveness of the combination therapies disclosed herein can be evaluated by determining the biological activity of immune cells present in a sample isolated from a treated individual. For example, the ability of immune cells isolated from the individual after treatment to destroy target cells, as determined using a cytotoxicity assay, can be used to evaluate the treatment efficacy. In some embodiments, the biological activity of immune cells present in a sample can be determined by measuring the expression and / or secretion of certain cytokines (e.g., IL-2 and IFNγ).

[0397] Unless otherwise indicated, the term "cancer vaccine" as used in the present invention refers to a "therapeutic cancer vaccine" administered to an individual with cancer for the treatment of cancer (and optionally for the prevention of recurrence of the cancer). In contrast, a "prophylactic cancer vaccine" is administered to an individual without cancer for the prevention of cancer or for reducing the risk of an 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 immunogenic compositions that comprise a pharmaceutically acceptable excipient and at least one tumor antigen, such as a tumor-specific antigen or a tumor-associated antigen. The terms "oncolytic virus" and "OV" as used in the present invention refer to a virus that infects and kills cancer cells. The death of cancer cells is the result of direct cell lysis and induction of anti-tumor immunity. In some embodiments, the "oncolytic virus" is a replication-competent virus that selectively replicates in cancer cells. In other embodiments, the "oncolytic virus" is a replication-defective virus that does not replicate in cancer cells due to genetic engineering or inactivation of the oncolytic virus (e.g., ultraviolet irradiation or heating).

[0398] In some embodiments, the tumor antigen comprises "common tumor antigens" that are shared by many cancers of the same type. Non-limiting examples of common tumor antigens are the breast cancer antigen HER2, the prostate cancer antigens PAP and PSA, and the melanoma antigens MART-1 and MAGE. In other embodiments, the tumor antigen comprises "neoantigens" that result from tumor-specific DNA alterations (e.g., somatic mutations). Thus, neoantigens typically have amino acid sequences that are not present in the normal mammalian genome (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. A tumor-specific neoantigen database (TSNAdb) is now available for free (Wu et al., Genomics Proteomics Bioinformatics 16:276-282, 2018).

[0399] Multiple techniques are applicable for identifying neoantigens included in cancer vaccines as part of combination therapies that include an activation threshold reducing agent (e.g., a Cbl-b inhibitor). For example, neoantigens can be identified by a method that includes isolating DNA from a tumor biopsy obtained from an individual, sequencing the DNA, and performing computational analysis of the sequences to identify one or more neoantigens (Aldous and Dong, Bioorg Med Chem, 26:2842-2849, 2018). In some embodiments, the computational analysis involves identifying peptides that are 8-11 amino acids in length, which are predicted to bind to at least one HLA allele expressed by tumor cells and that contain at least one missense mutation (Wu et al., Genomics Proteomics Bioinformatics 16:276-282, 2018). Neoantigens included in cancer vaccines are thought to be beneficial for overcoming tolerance and reducing the risk of autoimmunity.

[0400] Cancer vaccine platforms applicable to 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, F1000Research 2019, 8(F1000 FacultyRev):654, 2019).

[0401] In some embodiments, the tumor antigen of the cancer vaccine comprises at least one synthetic peptide or recombinant protein. In some embodiments, the synthetic peptide is at least 8 amino acids in length, and in certain embodiments, the synthetic peptide is less than 80 amino acids in length. In some embodiments, the tumor antigen comprises multiple synthetic peptides, or the tumor antigen comprises a synthetic peptide or recombinant protein that comprises the amino acid sequences of 2, 3, or more epitopes. An "epitope" is a part of an antigen that binds to an antibody or B cell receptor, or is presented by a major histocompatibility complex molecule (MHC class I or II) on the surface of a cell (e.g., a tumor cell or dendritic cell) for binding by a T cell receptor. In some embodiments, an epitope is a "linear epitope" (primary structure) composed of adjacent amino acids of a tumor antigen sequence. In some embodiments, an epitope is a "conformational epitope" (tertiary structure) composed of non-adjacent amino acids of a tumor antigen. In some embodiments, the tumor antigen comprises a recombinant protein that comprises both linear and conformational epitopes.

[0402] In some embodiments, the tumor antigen is encoded by a DNA or mRNA molecule. In some embodiments, the tumor antigen is encoded by the 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 BCG, a live culture preparation of the Bacillus of Calmette and Guerin (BCG) strain of Mycobacterium bovis, 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) and is suitable for the treatment and prevention of carcinoma in situ of the bladder, and the prevention of primary or recurrent stage Ta and / or T1 papillary tumors after transurethral resection.

[0403] In some embodiments, the microbial vector is a recombinant microbial vector, such as a recombinant viral vector or a recombinant bacterial vector. Recombinant viral vectors suitable for the combination therapy of the present invention include, but are not limited to, retroviruses, lentiviruses, adenoviruses, adeno-associated viruses, poxviruses, and herpesviruses (Chulpanova et al., Biomedicines, 6:94, 2018). In some embodiments, the microbial vector is a recombinant bacterial vector. Recombinant bacterial vectors suitable for the combination therapy of the present invention include, but are not limited to, Clostridium (e.g., Clostridium novyi), Listeria (e.g., Listeria monocytogenes), Pseudomonas (e.g., Pseudomonas aeruginosa), and Salmonella (e.g., Salmonella typhimurium) (Toussant et al., Expert Rev Vaccines, 12:1139-1154, 2013).

[0404] 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 a 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 sold by Dendreon Corporation (Seattle, WA) (Sipuleucel-T). Comprises lactated Ringer's solution (e.g., pharmaceutically acceptable excipient) and peripheral blood mononuclear cells (PBMCs) that have been activated by a PAP-GM-CSF fusion protein consisting of prostate acid phosphatase linked to granulocyte-macrophage colony-stimulating factor. Is an intravenous infusion approved by the Federal Drug Administration (FDA) for the treatment of asymptomatic or minimally symptomatic metastatic prostate cancer.

[0405] In some embodiments, the cancer vaccine comprises killed tumor cells. In some embodiments, the cancer vaccine comprises tumor cell lysates. In some embodiments, the cancer vaccine comprises APCs that have been contacted with tumor cell lysates.

[0406] Adjuvants for cancer vaccines suitable for the methods, medicaments, and uses of the present invention include, but are not limited to, adjuvants of FDA-approved licensed products. In particular, currently FDA-approved licensed product adjuvants include aluminum salts, monophosphoryl lipid A, oil-in-water emulsions (e.g., squalene-in-water emulsion MF59 or AS03), saponins, and CpG oligodeoxynucleotides.

[0407] Oncolytic viruses suitable for the methods, medicaments, and uses of the present invention include, but are not limited to, adenovirus, coxsackievirus, echovirus, fowlpoxvirus, herpes simplex virus, maraba virus, measles virus, myxoma virus, Newcastle disease virus, parvovirus, poliovirus, retrovirus, reovirus, Seneca Valley virus, Semiliki Forest virus, vaccinia virus, 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).

[0408] In some embodiments, the oncolytic virus is not genetically engineered (non-recombinant virus). In some embodiments, the non-recombinant virus is echovirus (e.g., Rigvir), Newcastle disease virus, parvovirus, reovirus, or Seneca Valley virus.

[0409] 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 a combination of one or more gene deletions and one or more gene insertions. In some embodiments, the recombinant virus has been genetically engineered to alter host cell specificity and / or tumor cell cytotoxicity. In some embodiments, the recombinant oncolytic 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., 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 transgene encoding a detectable marker (e.g., a fluorescent protein). Desirable anti-tumor responses include one or both of innate immune responses and adaptive immune responses.

[0410] In some embodiments, the recombinant oncolytic virus is a recombinant herpes simplex virus (HSV), such as HSV-1. In one embodiment, the recombinant oncolytic virus is also known as talimogene laherparepvec or T-VEC, and is marketed by Amgen Inc., Thousand Oaks, CA. is a recombinant HSV-1 that includes functional deletions of the ICP34.5 and ICP47 genes, and an insertion of a nucleic acid encoding human granulocyte macrophage colony-stimulating factor (GM-CSF). has been approved by the U.S. Food and Drug Administration (FDA) for the local treatment of unresectable cutaneous, subcutaneous, and nodal lesions in patients with recurrent melanoma by intralesional injection (intratumoral administration). In addition, has been approved by the European Medicines Agency (EMA) for the treatment of adults with unresectable melanoma with regional or distant metastases (stages IIIB, IIIC, or IVM1a). In particular, the EMA-approved product will be administered by intralesional injection (intratumoral administration) to cutaneous, subcutaneous, and / or nodal lesions that are visible, palpable, or detectable by ultrasound guidance.

[0411] In some embodiments, the recombinant oncolytic virus is a recombinant adenovirus, such as adenovirus serotype 5. In one embodiment, the recombinant adenovirus is Oncorine (H101), previously known as Onyx-015. Oncorine is an adenovirus serotype 5 that has been engineered by inactivating (functionally deleting) the viral E1B-55k and viral E3 genes. Oncorine has been approved by the National Medical Products Administration for the treatment of head and neck cancer in combination with chemotherapy (antineoplastic agent therapy).

[0412] In some embodiments, the recombinant oncolytic virus is a recombinant poxvirus. In some embodiments, the poxvirus is vaccinia virus or fowlpox virus. In some embodiments, the vaccinia virus is 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., antitumor 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 that has been engineered by inactivating (functionally deleting) the viral thymidine kinase gene and by 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 contains a functional deletion of the viral thymidine kinase and vaccinia growth factor genes, and an inserted 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).

[0413] Further embodiments of the combination therapies of the invention comprise 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 (antineoplastic agents), radiotherapy, and combinations thereof.

[0414] 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 can comprise an antibody or an antigen-binding fragment thereof. The antibody or fragment can be a monoclonal antibody (mAb), a human antibody, a humanized antibody, or a chimeric antibody, and can include a human constant region. In some embodiments, the human constant region is selected from the group consisting of IgGl, IgG2, IgG3, and IgG4 constant regions, and in certain embodiments, the human constant region is an IgGl 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.

[0415] In some embodiments, the chemotherapy comprises at least one anti-tumor agent (i.e., WHO ATC code L01). In some embodiments, the at least one anti-tumor agent is selected from the group consisting of cytotoxic antibiotics, plant alkaloids, antimetabolites, alkylating agents, other anti-tumor agents, and combinations thereof. For chemotherapy, the anti-tumor agent is a "drug" rather than a "therapeutic biological product".

[0416] In some embodiments, the radiotherapy is external beam radiotherapy. In other embodiments, the radiotherapy is brachytherapy. In some embodiments, the radiotherapy is ablative radiotherapy.

[0417] The term "biosimilar" or "biological similar" as used in the present invention refers to a biological product that is similar to a reference product approved by the Federal Drug Administration (FDA) but has no clinically meaningful differences in terms of safety and efficacy. For example, there may be differences in clinically inactive components between a biosimilar product and the reference product (e.g., differences in formulation excipients, minor differences in glycosylation, etc.). Clinically meaningful characteristics can be evaluated through pharmacokinetic and pharmacodynamic studies. In some embodiments, the biosimilar product is an interchangeable product as determined by the FDA. In some embodiments, the cancer vaccine is a biosimilar of an FDA-approved product.

[0418] IV. Compositions, Formulations, and Routes of Administration

[0419] The present invention encompasses pharmaceutical compositions of any compound or its salt or solvate disclosed in the present invention. Accordingly, the present invention includes pharmaceutical compositions comprising a Cbl-b inhibitor, wherein the Cbl-B inhibitor is a compound represented by formula (I), (I-a), (I-b), (I-A)-(I-J), (II-A)-(II-H), (III-A)-(III-H), or (IV-A)-(IV-H) disclosed in the present invention, 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, and a pharmaceutically acceptable excipient, such as a pharmaceutically acceptable vehicle or a pharmaceutically acceptable carrier. In some embodiments, the compound is a compound numbered 1-53 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.

[0420] The compounds and compositions disclosed in the present invention can be administered in any suitable form and by any suitable route to provide a sufficient level of the compound for treating a disease or disorder. In some embodiments, the Cbl-b inhibitor and / or the additional therapeutic agent is administered by enteral administration. In certain embodiments, the enteral administration is oral administration. In other embodiments, the Cbl-b inhibitor and / or the additional therapeutic agent is administered by parenteral administration. In certain embodiments, the parenteral administration is intratumoral injection. In certain embodiments, the parenteral administration is by a route selected from the group consisting of intravenous, intraperitoneal, and subcutaneous.

[0421] Suitable routes of administration include oral administration, enteral administration, parenteral administration including subcutaneous injection, intravenous injection, intraarterial injection, intramuscular injection, intrasternal injection, intraperitoneal injection, intralesional injection, intraarticular injection, intratumoral injection, or infusion techniques. The compounds and compositions can also be administered sublingually, mucosally, buccally, subcutaneously, spinally, epidurally, intracerebroventricularly, by inhalation (e.g., as an aerosol or spray), intranasally, vaginally, rectally, topically, or transdermally, or by a sustained release or extended release mechanism. The compounds and compositions can be administered in unit dosage forms containing conventional pharmaceutically acceptable carriers, excipients, adjuvants, and vehicles as needed. The compounds and compositions can be administered directly to a specific or affected organ or tissue. The compounds can be mixed with pharmaceutically acceptable carriers, excipients, adjuvants, and vehicles to form compositions suitable for the desired route of administration. In some embodiments, the compound can be mixed with one or both of an antigen and an adjuvant. In some embodiments, the antigen is a cancer antigen.

[0422] In certain embodiments disclosed by the present invention, particularly those embodiments in which the formulation is for injection or other parenteral administration, including the routes listed in the present invention, but also including any other administration routes described in the present invention (e.g., oral, enteral, gastric, etc.), the formulation recipes and formulations used in these methods are all sterile. Sterile pharmaceutical formulations are all mixed or formulated according to the pharmaceutical-grade sterilization standards known to those skilled in the art (Chapter 797, 1072, and 1211 of the United States Pharmacopeia; California Business and Professions Code 4127.7; 16 California Code of Regulations and Administrative Rules 1751, 21 United States Code of Federal Regulations 211). A "sterile" formulation is sterile, or free from or substantially free from all living microorganisms and their spores. Examples of methods for sterilizing pharmaceutical formulations include, but are not limited to, sterile filtration through a sterile filtration membrane, exposure to radiation (e.g., γ-radiation), and heat sterilization.

[0423] Oral administration is advantageous because of its ease of implementation and patient compliance. If the patient has difficulty swallowing, the drug can be introduced through a feeding tube, feeding syringe, or gastrostomy to complete enteral administration. The active compound and other co-administered agents (if any) can be enterally administered in any other pharmaceutically acceptable excipients suitable for a formulation for administration through a feeding tube, feeding syringe, or gastrostomy.

[0424] Intravenous administration can also be advantageously used to deliver the compound or composition to the bloodstream as quickly as possible, avoiding the need for absorption from the gastrointestinal tract.

[0425] The compounds and compositions useful for the purposes of 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, gums, ointments, cataplasms (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 in 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, where the prodrug undergoes conversion to a therapeutically effective form in the subject being treated.

[0426] In addition, the pharmaceutical preparation may include preservatives, solubilizers, stabilizers, reweting agents, emulsifiers, sweeteners, dyes, regulators, and salts, buffers, coating agents, or antioxidants for adjusting the osmotic pressure. The preparation containing the compound may also include other substances with valuable therapeutic properties. The pharmaceutical preparation can be prepared by known pharmaceutical methods. Other preparations and administration methods are known in the art. Suitable preparations can be found, for example, in Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins, 21st ed. (2005), which is incorporated herein by reference.

[0427] Injectable preparations, such as sterile injectable aqueous or oily suspensions, can be formulated according to methods known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a parenterally acceptable diluent or solvent, such as a propylene glycol solution. Acceptable vehicles and solvents that can be used are water, saline, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile nonvolatile oils are commonly used as solvents or suspending media. In view of this, any mild nonvolatile oil can be used, including synthetic monoglycerides or diglycerides. Additionally, fatty acids, such as oleic acid, can be used in the preparation of injectable preparations.

[0428] Solid dosage forms for oral administration may include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound can be mixed with at least one inert diluent (such as sucrose, lactose, talc, or starch). Such dosage forms may also include additional excipient substances in addition to the inert diluent, 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 can also be prepared with enteric coatings. Excipients acceptable for soft shell gel capsules are, for example, vegetable oils, waxes, fats, semi-solids, and liquid polyols, etc.

[0429] Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs, which contain inert diluents commonly used in the art, such as water. Such compositions may also include additional agents, such as wetting agents, emulsifiers, and suspending agents, cyclodextrins, as well as sweeteners, flavoring agents, and aromatic agents. Alternatively, if appropriate, the compound can also be administered in the form of the pure compound.

[0430] The compounds and compositions may 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 from single or multiple layers of 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 in the present invention, 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 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, N.W., p. 33 et seq (1976).

[0431] The amount of the active ingredient that can be combined with the carrier material to produce a single dosage form can vary depending on the patient to whom the active ingredient is administered and the particular 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 surface area, body mass index (BMI), general health, gender, and diet; the time and route of administration employed; the rate of excretion; and the drug combination used (if any). The compounds can be administered in unit dosage formulations. The selected pharmaceutical unit doses are prepared and administered to provide a sufficient concentration of the drug in the patient, subject, or individual.

[0432] Although the compounds for use in 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 compounds for use in the present invention, the additional active agent can generally be used in a therapeutically effective amount as shown 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 in amounts determined empirically for each patient.

[0433] Combinations of two or more of the compounds and compositions disclosed herein may also be used. The two or more compounds or compositions may be mixed together shortly before administration and administered together. The two or more compounds or compositions may be administered simultaneously by the same route of administration or by different routes of administration. The two or more compounds or compositions may be administered sequentially by the same route of administration or by different routes of administration. In one embodiment, a kit form may contain two or more compounds or compositions as separate forms of the compounds or compositions, with printed or electronic instructions for administration as a mixture of the compounds or compositions, as simultaneous administration of the separate compounds or compositions, or as sequential administration of the separate compounds or compositions. When three or more compounds or compositions are administered, they may be administered as a mixture of the compounds or compositions, as separate compounds or compositions administered simultaneously, as separate compounds or compositions administered sequentially, as separate compounds or compositions where two or more may be administered simultaneously and the remainder administered sequentially before or after the simultaneous administration, or in any other possible combination of mixed, simultaneous, and sequential administration.

[0434] On the one hand, the compounds disclosed in the present invention can be in a purified form, and the present invention discloses a composition comprising the compound in the purified form. The present invention provides a composition comprising the compound disclosed in the present invention or a salt thereof, such as a composition of a substantially pure compound. In some embodiments, the composition comprising the compound disclosed in the present invention or a salt thereof is in a substantially pure form. In one variant, "substantially pure" means a composition comprising no more than 35% impurities, where the impurities represent compounds other than the compound (or compounds, if a combination of compounds is used) administered in the composition, or salts or solvates of this compound (or compounds, if a combination is used). The weight of any added vehicle, carrier or excipient is not included in such a 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 1 means a composition comprising no more than 35% impurities, where 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 variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, wherein the composition comprises no more than 20% impurities. In yet another variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, wherein the composition comprises no more than 10% impurities. In a further variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, wherein the composition comprises no more than 5% impurities. In another variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, wherein the composition comprises no more than 3% impurities. In yet another variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, wherein the composition comprises no more than 1% impurities. In a further variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, wherein the composition comprises no more than 0.5% impurities. In other variants, a composition of a substantially pure compound means that the composition comprises no more than 15%, no more than 10%, no more than 5%, no more than 3%, or no more than 1% impurities. The impurities can be compounds in a stereochemical form different from the desired stereochemical form. For example, a composition of a substantially pure (S)-compound means that the composition comprises 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 compound. Alternatively, the "enantiomeric excess (ee)" used in the present invention refers to a dimensionless molar ratio that describes 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 one enantiomer is not in excess relative to the other enantiomer).As a further example, an enantiomeric excess of 99 would represent an almost stereopure enantiomeric compound (i.e., one enantiomer is in large excess relative to the other enantiomer). The enantiomeric excess percentage, %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. In addition, the "diastereomeric excess (de)" used in the present invention 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 an almost stereopure diastereomeric compound (i.e., one diastereomer is in large excess relative to the other diastereomer). The diastereomeric excess can be calculated by a method similar to that for ee. As will be understood by those skilled in the art, de is usually reported as a percentage de (%de). %de can be calculated in a manner similar to %ee.

[0435] In some cases, the present invention provides a composition comprising a cell population, the cell population comprising modified immune cells, such as those modified immune cells described in the present invention or generated by the methods disclosed in the present invention. In some embodiments, the composition comprises a cell population, the cell population comprising modified immune cells that have been contacted with or are in contact with a Cbl-b inhibitor or a composition thereof as described in the present invention. In some embodiments, the modified immune cells have been contacted with or are in contact with an anti-CD3 antibody alone. In some embodiments, the modified immune cells have been contacted with or are in contact with a combination of an anti-CD28 antibody and an anti-CD3 antibody. The composition provided by the present invention comprising a cell population containing the modified immune cells described in the present invention may further comprise a pharmaceutically acceptable excipient.

[0436] In some cases, the present invention also provides a cell culture composition comprising a cell population, the cell population comprising the immune cells described in the present invention and a Cbl-b inhibitor. 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.

[0437] The modified immune cells or compositions of the present invention, such as compositions or pharmaceutical compositions comprising a cell population containing the modified immune cells, can be provided in a suitable container. Suitable containers include, for example, bottles, vials (e.g., dual-chamber vials), syringes (e.g., single-chamber or dual-chamber syringes), bags (e.g., intravenous infusion bags), and tubes (e.g., test tubes). The container can be formed from a variety of materials (e.g., glass or plastic).

[0438] In some embodiments, a composition (e.g., a cell culture composition) comprising a cell population containing the modified immune cells of the present invention can be provided in a culture vessel. Culture vessels provided by the present invention 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).

[0439] The present invention also provides the compositions of the present invention for any use described herein. In some embodiments, the compositions of the present invention are used to prepare a medicament for treating or preventing a disease or disorder associated with Cbl-b activity. In some embodiments, the compositions of the present invention are used to prepare a medicament for treating cancer.

[0440] The present invention includes pharmaceutical compositions of any compound or its salt or solvate disclosed in the present invention for use in combination with a cancer vaccine. Accordingly, the present disclosure 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 its "a" and "b" variants) of International Patent Application WO 2019 / 148005, or a compound represented by any of Formula (I-A), 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 or a mixture of stereoisomers thereof. In addition, the present invention includes pharmaceutical compositions of any compound or its salt or solvate disclosed in the present invention for use in combination with an oncolytic virus. Accordingly, the present disclosure includes pharmaceutical compositions 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 a compound represented by any of Formula (I-A), 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 or a mixture of stereoisomers thereof. The composition may further comprise a pharmaceutically acceptable excipient, such as a pharmaceutically acceptable vehicle or a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition comprises both a small molecule Cbl-b inhibitor and a cancer vaccine. 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 Cbl-b inhibitors disclosed in the following patent applications: Compound 1-53 of U.S. Patent Application No. 62 / 866,914 or the compound represented by Formula (I) therein; Compound 1-53 of U.S. Patent Application No. 62 / 880,285 or the compound represented by Formula (I) therein; 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.

[0441] On the one hand, the pharmaceutically acceptable salt is an acid addition salt, such as a salt formed with an inorganic acid or an organic acid.

[0442] The compounds, vaccines, and compositions disclosed in the present invention can all be administered in any suitable form and by any suitable route, which will provide a sufficient level of the compound, vaccine, or composition for treating a disease or disorder. In some embodiments, the Cbl-b inhibitor and / or the cancer vaccine are administered by enteral administration. The compounds, oncolytic viruses, and compositions disclosed in the present invention can all be administered in any suitable form and by any suitable route, which will provide a sufficient level of the compound, oncolytic virus, or composition for treating a disease or disorder. 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.

[0443] Suitable routes of administration include oral administration, enteral administration, parenteral administration including subcutaneous injection, intravenous injection, intra-arterial injection, intramuscular injection, intrasternal injection, intraperitoneal injection, intralesional injection, intra-articular injection, intratumoral injection, or infusion techniques. The compounds, vaccines, and compositions can also be administered sublingually, mucosally, buccally, subcutaneously, spinally, epidurally, intracerebroventricularly, by inhalation (e.g., as an aerosol or spray), intranasally, vaginally, rectally, topically, or transdermally, or by sustained release or extended release mechanisms. The compounds, vaccines, and compositions can be administered as unit dosage formulations containing conventional pharmaceutically acceptable carriers, excipients, adjuvants, and vehicles as needed. The compounds, vaccines, and compositions can be administered directly to a particular or affected organ or tissue. The compounds and / or vaccines can be mixed with pharmaceutically acceptable carriers, excipients, adjuvants, and vehicles to form a composition suitable for the desired route of administration. The compounds, oncolytic viruses, and compositions can also be administered sublingually, mucosally, buccally, subcutaneously, spinally, epidurally, intracerebroventricularly, by inhalation (e.g., as an aerosol or spray), intranasally, vaginally, rectally, topically, or transdermally, or by sustained release or extended release mechanisms. The compounds, oncolytic viruses, and compositions can be administered as unit dosage formulations containing conventional pharmaceutically acceptable carriers, excipients, adjuvants, and vehicles as needed. The compounds, oncolytic viruses, and compositions can be administered directly to a particular or affected organ or tissue. The compounds and / or oncolytic viruses can be mixed with pharmaceutically acceptable carriers, excipients, adjuvants, and vehicles to form a composition suitable for the desired route of administration.

[0444] In certain embodiments disclosed herein, particularly those embodiments in which the formulation is for injection or other parenteral administration, including the routes listed herein but also including any other route of administration described herein (e.g., oral, enteral, gastric, etc.), except for the presence of microbial vectors in certain cancer vaccines, the formulation and the preparation used in these methods are sterile. To prepare such a formulation, all components of the formulation or the preparation are prepared in a sterile manner prior to combination with the microbial vector. In certain embodiments disclosed herein, particularly those embodiments in which the formulation is for injection or other parenteral administration, including the routes listed herein but also including any other route of administration described herein (e.g., oral, enteral, gastric, etc.), except for the presence of oncolytic viruses, the formulation and the preparation used in these methods are sterile. To prepare such a formulation, all components of the formulation or the preparation are prepared in a sterile manner prior to combination with the oncolytic virus. Sterile pharmaceutical preparations are mixed or formulated according to pharmaceutical grade sterilization standards known to those skilled in the art (Chapter 797, 1072, and 1211 of the United States Pharmacopeia; California Business and Professions Code 4127.7; 16 California Code of Regulations and Administrative Rules 1751, 21 United States Code of Federal Regulations 211). A "sterile" preparation is sterile, or free of or substantially free of all living microorganisms and their spores. Examples of methods for sterilizing pharmaceutical preparations include, but are not limited to, sterile filtration through a sterile filter membrane, exposure to radiation (e.g., gamma radiation), and heat sterilization.

[0445] Oral administration is advantageous because of its ease of implementation and patient compliance. If the patient has difficulty swallowing, the drug can be introduced through a feeding tube, feeding syringe, or gastrostomy to effect enteral administration. The active compound, vaccine, or composition, and any other co-administered agents (if present), can be enterally administered in any other pharmaceutically acceptable excipient suitable for administration via a feeding tube, feeding syringe, or gastrostomy. The active compound, oncolytic virus, or composition, and any other co-administered agents (if present), can be enterally administered in any other pharmaceutically acceptable excipient suitable for administration via a feeding tube, feeding syringe, or gastrostomy.

[0446] Intravenous administration can also be advantageously used to deliver the compound, vaccine, or composition to the bloodstream as quickly as possible, obviating the need for absorption from the gastrointestinal tract. Intravenous administration can also be advantageously used to deliver the compound, oncolytic virus, or composition to the bloodstream as quickly as possible, obviating the need for absorption from the gastrointestinal tract.

[0447] The compounds, vaccines, and compositions useful for the purposes of the present invention can be in solid form, liquid form, aerosol form, or in the form of tablets, pills, cachets, capsules (e.g., hard gelatin capsules or soft elastic gelatin capsules), powder mixtures, granules, injections, solutions, suppositories, enemas, colon lavages, emulsions, dispersions, food premixes, lozenges, troches, pastilles, gums, ointments, cataplasms (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 in other forms suitable for the route of administration. The compounds, vaccines, and compositions can also be administered in liposomal formulations. The compounds, oncolytic viruses, and compositions useful for the purposes of the present invention can be in solid form, liquid form, aerosol form, or in the form of tablets, pills, cachets, capsules (e.g., hard gelatin capsules or soft elastic gelatin capsules), powder mixtures, granules, injections, solutions, suppositories, enemas, colon lavages, emulsions, dispersions, food premixes, lozenges, troches, pastilles, gums, ointments, cataplasms (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 in other forms suitable for the route of administration. The compounds, oncolytic viruses, 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.

[0448] In addition, pharmaceutical formulations can include preservatives, solubilizers, stabilizers, rewetting agents, emulsifiers, sweeteners, dyes, regulators, and salts, buffers, coating agents, or antioxidants for adjusting osmotic pressure. Preparations containing the compounds, vaccines, or compositions can also include other substances having valuable therapeutic properties. Preparations containing the compounds, oncolytic viruses, or compositions can also include other substances having valuable therapeutic properties. Pharmaceutical formulations can be prepared by known pharmaceutical methods. Other formulations and methods of administration are known in the art. Suitable formulations can be found, for example, in Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins, 21st ed. (2005), which is incorporated herein by reference.

[0449] Injectable preparations, such as sterile injectable aqueous or oleaginous suspensions of small molecule Cbl-b inhibitors, can be formulated according to methods known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a parenterally acceptable diluent or solvent, such as a propylene glycol solution. Acceptable vehicles and solvents that can be used are water, saline, Ringer's solution, and isotonic sodium chloride solution. In addition, a sterile, nonvolatile oil is commonly used as a solvent or suspending medium. In view of this, any mild nonvolatile oil can be used, including synthetic monoglycerides or diglycerides. Additionally, fatty acids, such as oleic acid, can be used in the preparation of injectable preparations.

[0450] Solid dosage forms for oral administration can include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound, vaccine, or composition can be admixed with at least one inert diluent (such as sucrose, lactose, talc, or starch). In such solid dosage forms, the active compound, oncolytic virus, or composition can be admixed with at least one inert diluent (such as sucrose, lactose, talc, or starch). Such dosage forms can also contain additional excipient substances in addition to the inert diluent, for example, lubricants such as magnesium stearate. In the case of capsules, tablets, and pills, the dosage form can also contain buffering agents. Tablets and pills can also be prepared with enteric coatings. Excipients acceptable for soft shell gel capsules are, for example, vegetable oils, waxes, fats, semisolids, and liquid polyols, etc.

[0451] Liquid dosage forms for oral administration can include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs, which contain inert diluents commonly used in the art, such as water. Such compositions can also contain additional agents, such as wetting agents, emulsifying agents, and suspending agents, cyclodextrins, as well as sweetening agents, flavoring agents, and aromatic agents. Alternatively, if appropriate, the compound can also be administered in the form of the pure compound.

[0452] The compounds, vaccines and compositions may also be administered in the form of liposomes. The compounds, oncolytic viruses and compositions may 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 from single or multiple layers of 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 or vaccines disclosed in the present invention, the compositions of the present invention in liposome form may also contain stabilizers, preservatives, excipients, etc. In addition to the compounds or oncolytic viruses disclosed in the present invention, 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, N.W., p. 33 et seq (1976).

[0453] The amount of the active ingredient that can be combined with the carrier material to produce a single dosage form may 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 or vaccine used; the patient's age, weight, body surface area, body mass index (BMI), general health, gender and diet; the time and route of administration employed; the excretion rate; and the drug combination used (if any). The compounds, vaccines and compositions may be administered in unit dosage formulations. 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 or oncolytic virus used; the patient's age, weight, body surface area, body mass index (BMI), general health, gender and diet; the time and route of administration employed; the excretion rate; and the drug combination used (if any). The compounds, oncolytic viruses and compositions may be administered in unit dosage formulations. The selected pharmaceutical unit doses are prepared and administered to provide a sufficient concentration of the drug in a patient, subject or individual.

[0454] Although the compounds, vaccines, and compositions for the uses described in the present invention may be administered as the sole active agent, they may also be used in combination with one or more other agents. When an additional active agent is used in combination with the compounds, vaccines, or compositions for the uses described in the present invention, the additional active agent may generally be used in a therapeutically effective amount as shown in the Physicians' Desk Reference (PDR), 71st Edition (2017), which is incorporated herein by reference, or in such therapeutically useful amounts as are known to those of ordinary skill in the art, or in amounts determined empirically for each patient. Although the compounds, oncolytic viruses, and compositions for the uses described in the present invention may be administered as the sole active agent, they may also be used in combination with one or more other agents. When an additional active agent is used in combination with the compounds, oncolytic viruses, or compositions for the uses described in the present invention, the additional active agent may generally be used in a therapeutically effective amount as shown in the Physicians' Desk Reference (PDR), 71st Edition (2017), which is incorporated herein by reference, or in such therapeutically useful amounts as are known to those of ordinary skill in the art, or in amounts determined empirically for each patient.

[0455] Combinations of two or more of the compounds, vaccines, and compositions disclosed in the present invention 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 form may contain two or more compounds, vaccines, or compositions as separate forms of the compound, vaccine, or composition, with printed or electronic instructions for administration as a mixture of the compound, vaccine, or composition, as separate compounds, vaccines, or compositions administered simultaneously, or as separate compounds, vaccines, or compositions administered sequentially. When three or more compounds, vaccines, or compositions are administered, they may be administered as a mixture of the compound, vaccine, or composition, 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 remainder administered sequentially before or after the simultaneous administration, or in any other possible combination of mixed administration, simultaneous administration, and sequential administration. Combinations of two or more of the 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 shortly before administration and administered together. 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, a kit form may contain two or more compounds, oncolytic viruses, or compositions as separate forms of the compound, oncolytic virus, or composition, with printed or electronic instructions for administration as a mixture of the compound, oncolytic virus, or composition, as separate compounds, oncolytic viruses, or compositions administered simultaneously, or as separate compounds, oncolytic viruses, or compositions administered sequentially. When three or more compounds, oncolytic viruses, or compositions are administered, they may be administered as a mixture of the compound, oncolytic virus, or composition, as separate compounds, oncolytic viruses, or compositions administered simultaneously, as separate compounds, oncolytic viruses, or compositions administered sequentially, as separate compounds, oncolytic viruses, or compositions in which two or more may be administered simultaneously and the remainder administered sequentially before or after the simultaneous administration, or in any other possible combination of mixed administration, simultaneous administration, and sequential administration.

[0456] The compounds disclosed in the present invention that can be used in the pharmaceutical compositions and methods of the present invention can, in one aspect, be in a purified form, and the present invention discloses compositions comprising the compounds in such purified form. The present invention provides compositions comprising a compound or a salt thereof disclosed in the present invention, such as compositions of substantially pure compounds. In some embodiments, the compositions comprising a compound or a salt thereof disclosed in the present invention are in a substantially pure form. In one variant, "substantially pure" means a composition comprising no more than 35% impurities, where the impurities represent compounds other than the compound (or compounds, if a combination of compounds is used) administered in the composition, or salts or solvates of such compound (or compounds, if a combination is used). The weight of any added vehicle, carrier, or excipient is not included in such 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 1 means a composition comprising no more than 35% impurities, where 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, where the composition comprises no more than 25% impurities. In another variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, where the composition comprises no more than 20% impurities. In yet another variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, where the composition comprises no more than 10% impurities. In a further variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, where the composition comprises no more than 5% impurities. In another variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, where the composition comprises no more than 3% impurities. In yet another variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, where the composition comprises no more than 1% impurities. In a further variant, the present invention provides a composition of a substantially pure compound or its salt or solvate, where the composition comprises no more than 0.5% impurities. In other variants, a composition of a substantially pure compound means the composition comprises no more than 15%, no more than 10%, no more than 5%, no more than 3%, or no more than 1% impurities. The impurities can be compounds of a stereochemical form different from the desired stereochemical form. For example, a composition of a substantially pure (S)-compound means the composition comprises no more than 15%, no more than 10%, no more than 5%, no more than 3%, or no more than 1% ...

Claims

1. A compound represented by formula (I): or a tautomer thereof, or a pharmaceutically acceptable salt thereof, wherein ， Yes Z 1 is CH or N; Z 2 is N; R 1 is -CF 3 or cyclopropyl; R 2 is -CF 3 or cyclopropyl; R 3 is H, C 1 -C 2 alkyl, or C 1 -C 2 haloalkyl; R 4 is a 4- to 8-membered heterocyclic group or a C 3 -C 6 cycloalkyl group wherein the heterocyclic group or cycloalkyl group is each optionally substituted by 1 to 5 R 6 groups; or R 3 and R 4 together with the carbon atom to which it is attached form a C 3 -C 5 cycloalkyl or 4- to 6-membered heterocyclic group, wherein the C 3 -C 5 cycloalkyl and 4- to 6-membered heterocyclic group are each optionally substituted by 1 to 5 R 6 groups; R 5 is H, C 1 -C 6 alkyl or C 1 -C 6 haloalkyl; Each R 6 is independently selected from C 1 -C 6 -alkyl, halogen, hydroxyl, -O(C 1 -C 6 -alkyl), -CN, C 1 -C 6 -alkyl-CN, C 1 -C 6 -alkyl-OH or C 1 -C 6 -haloalkyl; or two Rs attached to the same carbon atom 6 together with the carbon atom to which they are attached form a spiro C 3 -C 6 cycloalkyl or spiro 4- to 6-membered heterocyclic group; X is H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkyl-OH, C 1 -C 6 alkyl-CN, optionally substituted by 1 to 5 R 8 groups of C 3 -C 6 cycloalkyl, or is a 4- to 7-membered heterocyclic group, or a 5- to 8-membered heteroaryl group, wherein the heterocyclic group or heteroaryl group each optionally contains 1 to 2 additional heteroatoms selected from the group consisting of N, O, and S, and wherein the heterocyclic group or heteroaryl group each is optionally substituted with 1 to 5 R 8 groups; Each R 7 is independently H, C 1 -C 6 alkyl, C 1 -C 6 alkyl-OH, or C 1 -C 6 haloalkyl; and Each R 8 is independently halogen, C 1 -C 6 -alkyl, C 1 -C 6 -alkyl-CN, C 1 -C 6 -alkyl-OH, C 1 -C 6 -haloalkyl, -CN, oxo, or -O(C 1 -C 6 -alkyl).

2. The compound according to claim 1, wherein is 3. The compound according to claim 2, wherein Z 1 is CH.

4. The compound according to claim 3, wherein is 5. The compound according to claim 2, wherein Z 1 is N.

6. The compound according to claim 5, wherein is 7. The compound according to claim 1, wherein is 8. The compound according to claim 7, wherein is 9. The compound according to claim 8, wherein is 10. The compound according to any one of claims 1-9, wherein, R 3 is H, C 1 -C 2 alkyl, or C 1 -C 2 haloalkyl; R 4 is a 4- to 6-membered heterocyclic group or a C 4 -C 5 cycloalkyl group wherein the heterocyclic group or cycloalkyl group is each optionally substituted with 1 to 3 R 6 groups; or R 3 and R 4 together with the carbon atom to which it is attached forms a C 4 -C 5 cycloalkyl or 4- to 6-membered heterocyclic group, wherein C 4 -C 5 the cycloalkyl group and the 4- to 6-membered heterocyclic group are each optionally substituted with 1 to 3 R 6 groups.

11. The compound according to claim 10, wherein R 3 is H, -CH 3 , or -CF 3 ; and R 4 is cyclobutyl or or R 3 and R 4 together with the carbon atom to which they are attached form: wherein each group is each independently optionally substituted with 1 to 3 R 6 groups.

12. The compound according to claim 11, wherein R 3 and R 4 together with the carbon atom to which they are attached form 13. The compound according to any one of claims 1-9, wherein, Each R 6 is independently C 1 -C 3 alkyl, halogen, hydroxy, -O(C 1 -C 3 alkyl), -CN, C 1 -C 3 alkyl-CN, C 1 -C 3 alkyl-OH, or C 1 -C 3 haloalkyl; or two Rs attached to the same carbon atom 6 together with the carbon atom to which they are attached form a spiro C 3 -C 6 cycloalkyl or spiro 4- to 5-membered heterocyclic group.

14. The compound according to claim 13, wherein each R 6 is independently -CH 3 , F, hydroxy, -OCH 3 , -CN, -CH 2 CN, -CH 2 OH, or -CF 3 ; or two Rs attached to the same carbon atom 6 together with the carbon atom to which they are attached form a spirocyclopropyl group.

15. The compound according to any one of claims 1-9, wherein R 5 is H, C 1 -C 3 alkyl or C 1 -C 3 haloalkyl.

16. The compound according to claim 15, wherein R 5 is H, -CH 3 or -CHF 2 .

17. The compound according to any one of claims 1-9, wherein, X is H, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, C 1 -C 3 alkyl-OH, C 1 -C 3 alkyl-CN, or optionally C 8 substituted by 1 to 3 R 3 -C 5 cycloalkyl.

18. The compound according to claim 17, wherein X is H or -CH 3 .

19. The compound according to any one of claims 1-9, wherein X is 20. The compound according to claim 19, is a 4- to 6-membered heterocyclic group or a 5- to 6-membered heteroaryl group, wherein the heterocyclic group or heteroaryl group each optionally contains 1 to 2 additional heteroatoms selected from the group consisting of N, O, and S, and wherein the heterocyclic group or heteroaryl group each is optionally substituted with 1 to 5 R 8 groups.

21. The compound according to claim 20, wherein is a 4- to 5-membered heterocyclic group or a 5- to 6-membered heteroaryl group, wherein the heterocyclic group or heteroaryl group each optionally contains 1 additional heteroatom selected from the group consisting of N and O, and wherein the heterocyclic group or heteroaryl group each is optionally substituted with 1 to 5 R 8 groups.

22. The compound according to claim 19, wherein X is and Y is O, -CH 2 -, -CHR 8 -, or -C(R 8 ) 2 -.

23. The compound according to claim 22, wherein Y is O.

24. The compound according to claim 22, wherein Y is -CH 2 -, -CHR 8 -, or -C(R 8 ) 2 -.

25. The compound according to claim 19, wherein each R 7 is independently H, C 1 -C 3 alkyl, C 1 -C 3 alkyl-OH, or C 1 -C 3 haloalkyl.

26. The compound according to claim 25, wherein each R 7 is independently H, -CH 3 , -CH 2 OH, or -CF 3 .

27. The compound according to claim 19, wherein each R 8 is independently selected from halogen, C 1 -C 3 alkyl, C 1 -C 3 alkyl-CN, C 1 -C 3 alkyl-OH, C 1 -C 3 haloalkyl, -CN, oxo, or -O(C 1 -C 3 alkyl).

28. The compound according to claim 27, wherein each R 8 is independently F, -CH 3 , -CH 2 CH 3 , -CH 2 CN, -CH 2 OH, -CF 3 , -CN, oxo, or -OCH 3 .

29. A compound selected from or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

30. A pharmaceutical composition comprising the compound according to any one of claims 1-29 and a pharmaceutically acceptable excipient; or comprising the compound according to any one of claims 1-29 and one or both of an adjuvant and an antigen.

31. A non-disease treatment method for regulating the activity of immune cells in vitro or ex vivo, the method comprising contacting the immune cells with an effective amount of a Cbl-b inhibitor to regulate the activity of the immune cells, wherein the Cbl-b inhibitor is the compound according to any one of claims 1-29.

32. The method according to claim 31, wherein the immune cells comprise T cells, B cells, or natural killer (NK) cells.

33. The method according to claim 31 or 32, wherein the immune cells are isolated from a blood sample of a mammalian subject.

34. The method according to claim 31 or 32, wherein the immune cells are tumor-infiltrating lymphocytes (TIL), and the tumor-infiltrating lymphocytes (TIL) are isolated from a tumor of a mammalian subject suffering from cancer.

35. The method according to claim 31 or 32, wherein the immune cells comprise T cells, and wherein regulating the activity of the T cells comprises one or more of increased T cell activation, increased T cell proliferation, decreased T cell exhaustion, and decreased T cell tolerance.

36. The method according to claim 31 or 32, wherein the immune cells comprise NK cells, and wherein regulating the activity of the NK cells comprises increased NK cell activation.

37. The method according to claim 31 or 32, wherein the immune cells comprise B cells, and wherein regulating the activity of the B cells comprises increased B cell activation, optionally wherein the increased B cell activation comprises increased expression of CD69.

38. The method according to claim 31 or 32, wherein the immune cells are human immune cells.

39. The method according to claim 31 or 32, wherein the immune cells comprise a recombinant chimeric receptor.

40. The method according to claim 35, 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.

41. The method according to claim 40, wherein the cytokines comprise one or more selected from the group consisting of IL-2, IFN-γ, TNFα, and GM-CSF.

42. The method according to claim 40, wherein the T cell activation marker comprises one or more selected from the group consisting of CD25, CD69, and CTLA4.

43. The method according to claim 35, wherein the T cells are contacted with an anti-CD3 antibody alone or a combination of an anti-CD3 antibody and an anti-CD28 antibody; or it further comprises culturing the immune cells with IL-2 alone or a combination of IL-2 and an anti-CD3 antibody and / or an anti-CD28 antibody.

44. The method according to claim 36, wherein the increased NK cell activation comprises an increase in the production of cytokines.

45. The method according to claim 44, wherein the cytokine comprises one or more selected from the group consisting of IFN-γ, TNFα, and MIP1β.

46. The method according to claim 39, wherein the recombinant chimeric receptor is a chimeric antigen receptor.

47. A method for generating modified immune cells in vitro or ex vivo, comprising culturing a cell population comprising immune cells in the presence of an effective amount of a compound according to any one of claims 1-29 to regulate the activity of the immune cells, thereby generating the modified immune cells.

48. The method according to claim 47, which further comprises culturing the immune cells with an anti-CD3 antibody alone or a combination of an anti-CD3 antibody and an anti-CD28 antibody; or it further comprises culturing the immune cells with IL-2 alone or a combination of IL-2 and an anti-CD3 antibody and / or an anti-CD28 antibody; or it further comprises recovering the modified immune cells.

49. The method according to claim 47 or 48, wherein the immune cells are isolated from a blood sample of a mammalian subject.

50. The method according to claim 47 or 48, wherein the immune cells are tumor infiltrating lymphocytes (TILs), and the tumor infiltrating lymphocytes (TILs) are isolated from a tumor of a mammalian subject suffering from cancer.

51. The method according to claim 47 or 48, wherein 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.

52. The method according to claim 47 or 48, wherein the immune cells are tumor infiltrating lymphocytes (TILs).

53. The method according to claim 47 or 48, wherein the immune cells are human immune cells.

54. The method according to claim 47 or 48, wherein the immune cells or the modified immune cells comprise a recombinant chimeric receptor.

55. The method according to claim 47 or 48, wherein the modified 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.

56. The method according to claim 54, wherein the recombinant chimeric receptor is a chimeric antigen receptor.

57. Modified immune cells, which comprise a compound according to any one of claims 1-29.

58. The modified immune cell according to claim 57, wherein the immune cell is isolated from a blood sample of a mammalian subject, or wherein the immune cell is a tumor infiltrating lymphocyte (TIL), and the tumor infiltrating lymphocyte (TIL) is isolated from a tumor of a mammalian subject suffering from cancer; or wherein the modified immune cell is a T cell, and wherein the T cell exhibits one or more of increased T cell activation, increased T cell proliferation, decreased T cell exhaustion, and reduced T cell tolerance; or wherein the modified immune cell is a NK cell, and wherein the NK cell exhibits increased NK cell activation; or wherein the modified immune cell is a B cell, and wherein the B cell exhibits increased B cell activation, optionally wherein the increased B cell activation comprises increased expression of CD69; or wherein the modified immune cell is a human immune cell; or wherein the modified immune cell comprises a recombinant chimeric receptor.

59. The modified immune cell according to claim 58, 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.

60. The modified immune cell according to claim 59, wherein the cytokine comprises one or more selected from the group consisting of IL-2, IFN-γ, TNFα, and GM-CSF.

61. The modified immune cell according to claim 59, wherein the T cell activation marker comprises one or more selected from the group consisting of CD25, CD69, and CTLA4.

62. The modified immune cell according to claim 58, wherein the T cell is contacted with an anti-CD3 antibody alone or a combination of an anti-CD3 antibody and an anti-CD28 antibody; or wherein the T cell is contacted with IL-2 alone or a combination of IL-2 and an anti-CD3 antibody and / or an anti-CD28 antibody.

63. The modified immune cell according to claim 58, wherein the increased NK cell activation comprises increased production of cytokines.

64. The modified immune cell according to claim 63, wherein the cytokine comprises one or more selected from the group consisting of IFN-γ, TNFα, and MIP1β.

65. The modified immune cell according to claim 58, wherein the recombinant chimeric receptor is a chimeric antigen receptor.

66. A composition comprising a cell population, the cell population comprising the modified immune cell according to any one of claims 57-65.

67. The composition according to claim 66, further comprising a pharmaceutically acceptable excipient.

68. The composition according to claim 66, wherein the composition is in a suitable container.

69. The composition according to claim 68, wherein the suitable container is a vial, syringe, intravenous infusion bag, or tube. Use of a modified immune cell according to any one of claims 57 - 65 or a composition according to any one of claims 66 - 69 in the preparation of a medicament for modulating an immune response. Use of a modified immune cell according to any one of claims 57 - 65 or an effective amount of a composition according to any one of claims 66 - 69 in the preparation of a medicament for treating cancer responsive to inhibition of Cbl - b activity, wherein the cancer is responsive to inhibition of Cbl - b activity.

72. The use according to claim 71, wherein the cancer is a hematological cancer; or wherein the cancer is a non - hematological cancer.

73. The use according to claim 72, wherein the hematological cancer is lymphoma, leukemia, or myeloma; or wherein the non - hematological cancer is sarcoma, carcinoma, or melanoma. Use of a modified immune cell according to any one of claims 57 - 65 or a composition according to any one of claims 66 - 69 in the preparation of a medicament for inhibiting abnormal cell proliferation.

75. The use according to claim 74, wherein the abnormal cell proliferation is hyperplasia or cancer cell proliferation.

76. The use according to claim 75, wherein the cancer cells are from a hematological cancer; or wherein the cancer cells are from a non - hematological cancer.

77. The use according to claim 76, wherein the hematological cancer is lymphoma, leukemia, or myeloma; or wherein the non - hematological cancer is sarcoma, carcinoma, or melanoma. Use of a compound according to any one of claims 1 - 29 in the preparation of a medicament for modulating an immune response. Use of a compound according to any one of claims 1 - 29 in the preparation of a medicament for inhibiting Cbl - b activity. Use of a compound according to any one of claims 1 - 29 in the preparation of a medicament for treating cancer responsive to inhibition of Cbl - b activity.

81. The use according to claim 80, wherein the cancer is a hematological cancer.

82. The use according to claim 81, wherein the hematological cancer is lymphoma, leukemia, or myeloma.

83. The use according to claim 80, wherein the cancer is a non - hematological cancer.

84. The use according to claim 83, wherein the non - hematological cancer is sarcoma, carcinoma, or melanoma.

85. The use according to any one of claims 78 - 84, wherein the compound is administered enterally.

86. The use according to claim 85, wherein the enteral administration is oral administration.

87. The use according to any one of claims 78 - 84, wherein the compound is administered parenterally.

88. The use according to claim 87, wherein the parenteral administration is intratumoral administration.

89. The use according to any one of claims 80 - 84, further comprising administering an effective amount of the modified immune cells as defined in claim 57.

90. The use according to any one of claims 80 - 84, further comprising an effective amount of the composition as defined in claim 66 to treat the cancer.

91. Use of a compound according to any one of claims 1 - 29 in the manufacture of a medicament for inhibiting abnormal cell proliferation.

92. The use according to claim 91, wherein the abnormal cell proliferation is hyperplasia or cancer cell proliferation.

93. The use according to claim 92, wherein the cancer cells are from a hematological cancer.

94. The use according to claim 93, wherein the hematological cancer is lymphoma, leukemia, or myeloma.

95. The use according to claim 92, wherein the cancer cells are from a non - hematological cancer.

96. The use according to claim 95, wherein the non - hematological cancer is sarcoma, carcinoma, or melanoma.

97. The use according to any one of claims 91 - 96, wherein the compound is administered by enteral administration.

98. The use according to claim 97, wherein the enteral administration is oral administration.

99. The use according to any one of claims 91 - 96, wherein the compound is administered by parenteral administration.

100. The use according to claim 99, wherein the parenteral administration is intratumoral injection.

101. The use according to claim 99, the parenteral administration is by a route selected from the group consisting of intravenous, intraperitoneal, and subcutaneous.

102. The use according to claim 78, wherein modulating the immune response comprises one or more of increased T - cell activation, increased T - cell proliferation, decreased T - cell exhaustion, and decreased T - cell tolerance after administering the compound; or increased NK - cell activation after administering the compound; or increased B - cell activation after administering the compound, optionally wherein the increased B - cell activation comprises increased expression of CD69.

103. The use according to claim 102, wherein the increased T - cell activation comprises increased cytokine production; or wherein the increased T - cell activation comprises increased cell - surface expression of one or more T - cell activation markers.

104. The use according to claim 103, wherein the cytokine comprises one or more selected from the group consisting of IL - 2, IFN - γ, TNFα, and GM - CSF.

105. The use according to claim 103, wherein the T - cell activation marker comprises one or more selected from the group consisting of CD25, CD69, and CTLA4.

106. The use according to claim 102, wherein the increased NK - cell activation comprises increased cytokine production.

107. The use according to claim 106, wherein the cytokine comprises one or more selected from the group consisting of IFN - γ, TNFα, and MIP1β.

108. A cell culture composition comprising a cell population, said cell population comprising immune cells and the compound of claim 1.

109. The cell culture composition according to claim 108, wherein 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; or the immune cells are engineered immune cells comprising a recombinant chimeric receptor; or it further comprises an anti-CD3 antibody alone or a combination of an anti-CD3 antibody and an anti-CD28 antibody.

110. The cell culture composition according to claim 109, wherein the recombinant chimeric receptor is a chimeric antigen receptor.

111. The pharmaceutical composition according to claim 30, wherein the antigen is a cancer antigen.

112. An article comprising the pharmaceutical composition of claim 30, the modified immune cell of claim 57, the composition of claim 66, or the cell culture composition of claim 108.

113. The article according to claim 112, wherein the modified immune cell or cell culture composition is in a tube, dish, bag, multi-well plate, or flask.

114. The article according to claim 112, wherein the modified immune cell or pharmaceutical composition is in a vial, syringe, intravenous infusion bag, or tube.

115. A kit comprising the pharmaceutical composition of claim 30, the modified immune cell of claim 57, the composition of claim 66, or the cell culture composition of claim 108.

116. The kit according to claim 115, wherein the modified immune cell is in a tube, dish, bag, multi-well plate, or flask.

117. The kit according to claim 115, wherein the modified immune cell is in a vial, syringe, intravenous infusion bag, or tube.

118. Use of the compound according to any one of claims 1-29 in the preparation of a drug for treating or preventing a disease or disorder associated with Cbl-b activity.

119. Use of the compound according to any one of claims 1-29 in the preparation of a drug for treating cancer.

120. The use according to claim 119, wherein the cancer is a cancer associated with Cbl-b activity.

121. Use of the compound according to any one of claims 1-29 and an additional therapeutic agent in the preparation of a drug for treating cancer.

122. The use according to claim 121, wherein the compound and the additional therapeutic agent are administered sequentially in either order.

123. The use according to claim 121, wherein the compound and the additional therapeutic agent are administered simultaneously.

124. The use according to claim 121 or 122, wherein the compound is administered by enteral administration.

125. The use according to claim 124, wherein the enteral administration is oral administration.

126. The use according to claim 121 or 122, wherein the compound is administered by parenteral administration.

127. The use according to claim 126, wherein the parenteral administration is intratumoral injection.

128. The use according to claim 126, wherein the parenteral administration is by a route selected from the group consisting of intravenous, intraperitoneal, and subcutaneous.

129. The use according to claim 121 or 122, further comprising administering to the individual an effective amount of radiotherapy.

130. The use according to claim 121, wherein the additional therapeutic agent comprises an immune checkpoint inhibitor; or the additional therapeutic agent comprises an anti-tumor agent.

131. The use according to claim 130, wherein the immune checkpoint inhibitor is an antagonist of at least one inhibitory checkpoint molecule 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.

132. The use according to claim 130, wherein the immune checkpoint inhibitor is an antagonist of at least one inhibitory checkpoint molecule selected from the group consisting of: PD-1 (CD279), PD-L1 (CD274), and CTLA-4 (CD152).

133. The use according to claim 132, wherein the at least one inhibitory checkpoint molecule comprises PD-1.

134. The use according to claim 130, wherein the immune checkpoint inhibitor is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, and their biosimilars.

135. The use according to claim 132, wherein the at least one inhibitory checkpoint molecule comprises PD-L1.

136. The use according to claim 130, wherein the immune checkpoint inhibitor is selected from the group consisting of atezolizumab, avelumab, durvalumab, and their biosimilars.

137. The use according to claim 132, wherein the at least one inhibitory checkpoint molecule comprises CTLA-4.

138. The use according to claim 130, wherein the immune checkpoint inhibitor is selected from the group consisting of ipilimumab, tremelimumab, and their biosimilars.

139. The use according to claim 130, wherein the immune checkpoint inhibitor comprises an antibody or an antigen-binding fragment thereof, or wherein the antibody or fragment is human or humanized.

140. The use according to claim 130, wherein the anti-tumor agent is classified as one of the group consisting of cytotoxic antibiotics, plant alkaloids, antimetabolites, alkylating agents, and other anti-tumor agents.

141. The use according to claim 140, wherein the anti-tumor agent comprises a cytotoxic antibiotic.

142. The use according to claim 141, wherein 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.

143. The use according to claim 140, wherein the anti-tumor agent comprises a plant alkaloid.

144. The use according to claim 143, wherein the plant alkaloid is selected from the group consisting of: trabectedin, cabazitaxel, paclitaxel poliglumex, docetaxel, paclitaxel, demecolcine, teniposide, etoposide, vintafolide, vinflunine, vinorelbine, vindesine, vincristine, and vinblastine.

145. The use according to claim 140, wherein the anti-tumor agent comprises an antimetabolite.

146. The use according to claim 145, wherein the antimetabolite is a pyrimidine analogue.

147. The use according to claim 145, wherein the antimetabolite is a purine analogue.

148. The use according to claim 145, wherein the antimetabolite is a folic acid analogue.

149. The use according to claim 145, wherein 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.

150. The use according to claim 140, wherein the anti-tumor agent comprises an alkylating agent.

151. The use according to claim 150, wherein the alkylating agent is selected from the group consisting of dacarbazine, temozolomide, pipobroman, mitobronitol, etoglucid, uracil mustard, ranimustine, nimustine, fotemustine, streptozocin, semustine, lomustine, carmustine, carboquone, triaziquone, thiotepa, mannosulfan, treosulfan, busulfan, bendamustine, prednimustine, trofosfamide, ifosfamide, mechlorethamine, melphalan, chlorambucil, and cyclophosphamide.

152. The use according to claim 140, wherein the anti-tumor agent comprises other anti-tumor agents selected from the group consisting of platinum compounds, protein kinase inhibitors, and other agents.

153. The use according to claim 152, wherein the anti-tumor agent comprises a platinum compound.

154. The use according to claim 153, wherein the platinum compound is selected from the group consisting of cisplatin, carboplatin, oxaliplatin, satraplatin, and polyplatillen.

155. The use according to claim 152, wherein the anti-tumor agent comprises a protein kinase inhibitor.

156. The use according to claim 152, wherein the anti-tumor agent comprises other agents.

157. The use according to claim 129, wherein the radiotherapy is external beam radiotherapy.

158. The use according to claim 129, wherein the radiotherapy is brachytherapy.

159. The use according to claim 121, wherein the cancer is a hematological cancer; or wherein the cancer is a non-hematological cancer.

160. The use according to claim 159, wherein the hematological cancer is lymphoma, leukemia, or myeloma; or wherein the non-hematological cancer is carcinoma, sarcoma, or melanoma.

161. A method for generating an expanded population of tumor-infiltrating lymphocytes (TIL), the method comprising: (a) obtaining in vitro or ex vivo a biological sample comprising TIL from an individual with cancer who has received a compound of any one of claims 1-29 and optionally an additional therapeutic agent; and (b) culturing the TIL in a cell culture medium comprising at least one T cell growth factor to generate an expanded population of TIL.

162. The method according to claim 161, wherein the at least one T cell growth factor comprises IL-2; or 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 compound.

163. The method according to claim 161 or 162, wherein the cancer is a non-hematological cancer.

164. The method according to claim 163, wherein the non-hematological cancer is sarcoma, carcinoma, or melanoma.

165. A composition comprising the expanded population of TIL generated by the method of any one of claims 161-164, and a physiologically acceptable buffer.

166. The use of the composition according to claim 165 in the preparation of a medicament for the treatment of cancer.

167. The use according to claim 166, which further comprises administering a therapeutically effective amount of the compound.

168. The use of the compound of claim 1 and a vaccine in the preparation of a medicament for immunization.

169. The use according to claim 168, wherein the immunization is a method for treating cancer, which comprises: administering a therapeutically effective amount of a small molecule compound to cancer, and administering a therapeutically effective amount of a therapeutic cancer vaccine.

170. Use according to claim 169, wherein the compound and the cancer vaccine are administered sequentially; or wherein the compound and the cancer vaccine are administered simultaneously.

171. Use according to claim 169 or 170, wherein the cancer vaccine is an immunogenic composition comprising at least one tumor antigen and a pharmaceutically acceptable excipient.

172. Use according to claim 169 or 170, wherein the cancer vaccine comprises a microbial vector.

173. Use according to claim 172, wherein the microbial vector is TICE-BCG.

174. Use according to claim 169 or 170, wherein the cancer vaccine comprises killed cancer cells or cancer cell lysates.

175. Use according to claim 171, wherein the at least one tumor antigen comprises at least one synthetic peptide or recombinant protein; or the immunogenic composition further comprises an adjuvant.

176. Use according to claim 175, wherein the adjuvant comprises one or more components selected from the group consisting of aluminum salts, squalene, and saponins.

177. Use according to claim 171, wherein the immunogenic composition further comprises antigen-presenting cells (APCs).

178. Use according to claim 177, wherein the APCs are dendritic cells.

179. Use according to claim 170, wherein the cancer vaccine is PROVENGE.

180. Use according to claim 172, wherein the microbial vector is a recombinant viral vector or a recombinant bacterial vector.

181. Use of the compound and the oncolytic virus according to any one of claims 1-29 in the preparation of a medicament for the treatment of cancer.

182. Use according to claim 181, wherein the compound and the oncolytic virus are administered sequentially.

183. Use according to claim 181, wherein the compound and the oncolytic virus are administered simultaneously.

184. Use according to any one of claims 181-183, wherein the oncolytic virus is a virus selected from the group consisting of 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, vaccinia virus, and vesicular stomatitis virus.

185. Use according to any one of claims 181 - 183, wherein the oncolytic virus is a recombinant virus, and the recombinant virus comprises one or both of a functional deletion of at least one viral gene and an insertion of at least one transgene.

186. Use according to any one of claims 181 - 183, wherein the oncolytic virus is a transgenic adenovirus type 5.

187. Use according to any one of claims 181 - 183, wherein the oncolytic virus is a transgenic herpes simplex virus type 1 (HSV - 1).

188. Use according to claim 187, wherein the transgenic HSV - 1 is talimogene laherparepvec.

189. Use according to any one of claims 181 - 183, wherein the oncolytic virus is a transgenic vaccinia virus.

190. Use according to claim 189, wherein the transgenic vaccinia virus is pexastimogene devacirepvec.

191. Use according to any one of claims 181 - 183, wherein the oncolytic virus is a non - recombinant oncolytic virus.

192. Use according to claim 191, wherein the non - recombinant oncolytic virus is a virus selected from the group consisting of echovirus, Newcastle disease virus, parvovirus, reovirus, and Seneca Valley virus.

193. Use according to claim 185, wherein the recombinant virus comprises a functional deletion of at least one viral gene and an insertion of at least one transgene; or the transgene encodes human granulocyte - macrophage colony - stimulating factor (GM - CSF).

194. Use according to claim 181, wherein the cancer is a hematological cancer; or the cancer is a non - hematological cancer.

195. Use according to claim 194, wherein the hematological cancer is lymphoma, leukemia, or myeloma; or the non - hematological cancer is carcinoma, sarcoma, or melanoma.

196. Use according to claim 168, wherein the compound is selected from or its tautomer, its stereoisomer, or its pharmaceutically acceptable salt.

197. Use of the compound according to claim 1 and a therapeutic cancer vaccine or an oncolytic virus in the preparation of a medicament for treating cancer.

198. Use according to claim 197, wherein the compound can further reduce the co - stimulation requirement of immune cells; or the compound can further promote tumor immune surveillance; or the compound is a small - molecule Cbl - b inhibitor.

199. A pharmaceutical composition comprising a cancer vaccine and the compound of claim 1, optionally wherein the composition further comprises a pharmaceutically acceptable excipient.

200. A kit for treating cancer in vitro or ex vivo, the kit comprising: (a) The compound of claim 1; (b) A therapeutic cancer vaccine; (c) Instructions for treating cancer by administering an effective amount of the said compound and the said therapeutic cancer vaccine.

201. A kit for treating cancer in vitro or ex vivo, the kit comprising: (a) A pharmaceutical composition comprising the compound of claim 1 and a therapeutic cancer vaccine; and (b) Instructions for treating cancer by administering an effective amount of the said pharmaceutical composition comprising the said compound and the said therapeutic cancer vaccine.

202. The use according to claim 197, the composition according to claim 199, or the kit according to claim 201, wherein the cancer vaccine is an immunogenic composition comprising at least one tumor antigen and a pharmaceutically acceptable excipient.

203. The use according to claim 202, wherein the cancer vaccine comprises a microbial vector.

204. The use according to claim 203, wherein the microbial vector is TICE-BCG.

205. The use according to claim 197, the composition according to claim 199, or the kit according to claim 201, wherein the cancer vaccine comprises killed cancer cells or cancer cell lysates.

206. The use, composition, or kit according to claim 202, wherein the at least one tumor antigen comprises at least one synthetic peptide or recombinant protein.

207. The use, composition, or kit according to claim 202, wherein the immunogenic composition further comprises an adjuvant.

208. The use, composition, or kit according to claim 202, wherein the immunogenic composition further comprises antigen-presenting cells (APCs).

209. The use according to claim 208, wherein the APC is a dendritic cell.

210. The use, composition, or kit according to claim 202, wherein the cancer vaccine is PROVENGE.

211. The use, composition, or kit according to claim 207, wherein the adjuvant comprises one or more components selected from the group consisting of aluminum salts, squalene, and saponins.

212. The use, composition, or kit according to claim 203, wherein the microbial vector is a recombinant viral vector or a recombinant bacterial vector.

213. A pharmaceutical composition comprising an oncolytic virus and the compound of claim 1, optionally wherein the composition further comprises a pharmaceutically acceptable excipient.

214. A kit for treating cancer in vitro or ex vivo, the kit comprising: (a) The compound of claim 1; (b) An oncolytic virus; (c) Instructions for treating cancer by administering an effective amount of the said compound and the said oncolytic virus.

215. A kit for treating cancer in vitro or ex vivo, the kit comprising: (a) A pharmaceutical composition comprising the compound of claim 1 and an oncolytic virus; and (b) Instructions for treating cancer by administering an effective amount of the said pharmaceutical composition comprising the said compound and the said oncolytic virus.

216. The use according to claim 197 or 198, the composition according to claim 213, or the kit according to claim 214 or 215, wherein the oncolytic virus is a virus selected from the group consisting of: 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, vaccinia virus, and vesicular stomatitis virus.

217. The use according to claim 197 or 198, the composition according to claim 213, or the kit according to claim 214 or 215, wherein the oncolytic virus is a recombinant virus, and the recombinant virus comprises one or both of a functional deletion of at least one viral gene and an insertion of at least one transgene.

218. The use according to claim 197 or 198, the composition according to claim 213, or the kit according to claim 214 or 215, wherein the oncolytic virus is a transgenic adenovirus type 5.

219. The use according to claim 197 or 198, the composition according to claim 213, or the kit according to claim 214 or 215, wherein the oncolytic virus is a transgenic herpes simplex virus type 1 (HSV-1).

220. The use, composition or kit according to claim 219, wherein the transgenic HSV-1 is talimogene laherparepvec.

221. The use according to claim 197 or 198, the composition according to claim 213, or the kit according to claim 214 or 215, wherein the oncolytic virus is a transgenic vaccinia virus.

222. The use, composition or kit according to claim 221, wherein the transgenic vaccinia virus is pexastimogene devacirepvec.

223. The use according to claim 197 or 198, the composition according to claim 213, or the kit according to claim 214 or 215, wherein the oncolytic virus is a non-recombinant oncolytic virus.

224. The use, composition or kit according to claim 223, wherein the non-recombinant oncolytic virus is a virus selected from the group consisting of echovirus, Newcastle disease virus, parvovirus, reovirus, and Seneca Valley virus.

225. The use, composition, or kit according to claim 217, wherein the recombinant virus comprises a functional deletion of at least one viral gene and the insertion of at least one transgene; or the transgene encodes human granulocyte macrophage colony-stimulating factor (GM-CSF).

226. The use according to claim 197, the composition according to claim 199 or 213, or the kit according to any one of claims 200, 201, 214 or 215, wherein the cancer is a hematological cancer; or the cancer is a non-hematological cancer.

227. The use, composition, or kit according to claim 226, wherein the hematological cancer is lymphoma, leukemia, or myeloma; or the non-hematological cancer is carcinoma, sarcoma, or melanoma.

228. The use according to claim 197, the composition according to claim 199 or 213, or the kit according to claim 200, 201, 214 or 215, wherein the compound is selected from or a tautomer, a stereoisomer, or a pharmaceutically acceptable salt thereof. The use of one or more therapeutic cells and the compound according to claim 1 in the preparation of a medicament for treating a disease by cell therapy, wherein the treatment of the therapeutic cells is enhanced by combination with the compound.

Images