TW202327610A - Improved treatments for advanced/metastatic cancers with checkpoint inhibitor resistance or resistance susceptibility - Google Patents

Abstract

This invention is in the area of improved therapeutic methods for difficult to treat locally advanced and/or metastatic cancers in patients whose cancer has advanced while being treated with an immune checkpoint inhibitor due to the development of resistance to the inhibitory effects of the immune checkpoint inhibitor or whose cancer is susceptible to the development of resistance to the effects of an immune checkpoint inhibitor. The methods of the present invention are particularly suitable for a select group of hard-to-treat patients with advanced/metastatic triple negative breast cancer (TNBC), recurrent or metastatic non-small cell lung cancer (NSCLC), advanced or metastatic and unresectable colorectal cancer and locally advanced or metastatic urothelial carcinoma, and provides extended progression free survival (PFS) and/or increased overall survival (OS) in these patient populations.

Description

Improved treatment of advanced/metastatic cancer with checkpoint inhibitor resistance or resistance susceptibility

The present invention is in the field of improved treatments for refractory locally advanced and/or metastatic cancers in patients whose cancers are being treated with immune checkpoint inhibitors while at the same time due to the presence of immune checkpoint inhibitors Resistance to inhibitory effects has progressed, or the cancer is predisposed to become resistant to the effects of immune checkpoint inhibitors. The methods of the invention may be particularly applicable to selected groups of patients with, inter alia, advanced/metastatic breast cancer (including triple negative breast cancer (TNBC)), recurrent or metastatic colorectal cancer, recurrent or metastatic non-small cell lung cancer (NSCLC) and refractory patients with locally advanced or metastatic urothelial carcinoma, and can provide prolonged progression-free survival (PFS) and/or increased overall survival (OS) in these patient populations.

Arguably the most important achievement in cancer therapy in the past decade has been the introduction of blocking immune checkpoints cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), programmed cell death protein-1 (PD-1) and programmed death ligand -1 (PD-L1) T cell-targeting immunomodulator. Both CTLA-4 and PD-1 function to physiologically inhibit unrestricted cytotoxic T effector functions. CTLA-4 (CD 152) is of the B7/CD28 family and mediates immunosuppression by indirectly attenuating signaling through the co-stimulatory receptor CD28. PD-1 is an inhibitory transmembrane protein expressed on T cells, B cells, natural killer cells (NK) and myeloid-derived suppressor cells (MDSC). PD-L1 is expressed on the surface of various tissue types, including many tumor cells and hematopoietic cells. Blocking the PD-1/PDL-1 pathway enhances anti-tumor T cell reactivity and promotes immune control of cancerous cells.

In 2011, ipilimumab (a human IgG1 k anti-CTLA-4 monoclonal antibody sold under the brand name YERVOY®) was approved by the Food and Drug Administration (FDA) in the United States (US) For the treatment of unresectable or metastatic melanoma. Ipilimumab is the first and only CTLA-4 inhibitor approved by the US Food and Drug Administration.

Since USFDA approved ipilimumab in 2011, the following 6 immune checkpoint inhibitors (ICI) have been approved for cancer therapy: PD-1 inhibitor nivolumab (OPDIVO ^® ), Pembrolizumab (KEYTRUDA ^® ), cemiplimab (LIBTAYO ^® ) and the PD-L1 inhibitor atezolizumab (TECENTRIQ ^® ), avelumab (avelumab) (BEVENCIO ^® ) and durvalumab (IMFINZI ^® ). Immune checkpoint inhibitors (ICIs) have become some of the most widely prescribed anticancer therapies. The importance of these remarkable breakthroughs in cancer treatment was recognized in 2018 when James Allison and Tasuku Honjo, two researchers credited with pioneering the use of immune checkpoint inhibitors to treat cancer, were awarded the Nobel Prize in Medicine (Nobel Prize). Prize in Medicine) (see Ledford, Heidi et al., Cancer immunologists scoop medicine Nobel prize. Nature, Vol. 562, Issue 7725, Oct. 2018, pp. 20+).

The effectiveness of these agents in cancer treatment has established immunotherapy as an alternative in cancer treatment. In some solid tumors, such as metastatic melanoma and non-small cell lung cancer, immunotherapy is the first-line treatment. These targeted immunotherapies have provided improved outcomes in many difficult-to-treat cancers. For example, in 2019, the USFDA and the European Medicines Agency (EMA) accelerated approval of atezolizumab (TECENTRIQ ^{® )} , a PD-L1 blocking antibody (immune checkpoint inhibitor [ICI]) ) in combination with nab-paclitaxel (nab-paclitaxel) for PD-L1 positive locally advanced/metastatic TNBC patients (see TECENTRIQ ^® drug insert). USFDA accelerated approval is based on clinical trial results indicating improved progression-free survival (PFS) in patients with PD-L1-positive TNBC (hazard ratio [HR]: 0.60 [0.48, 0.77]; p<0.0001; median: 7.4 months vs .4.8 months). Additionally, the PD-L1-positive subgroup survived an average of 25 months when treated with atezolizumab plus nab-paclitaxel, compared with an average of 18 months when given placebo plus nab-paclitaxel. The improvement in efficacy observed by adding atezolizumab to nab-paclitaxel was associated with a relatively low frequency of immune-related adverse events that can cause significant morbidity and mortality.

Pembrolizumab (KEYTRUDA ^® ) monotherapy has also demonstrated improved objective response rates (ORR) between 18.5% and 21.4% in patients with PD-L1-positive TNBC tumors compared to PD-L1-negative Response rates in tumor patients ranged from 5.3% to 9.6%. Similarly, a statistically significant and clinically meaningful improvement in PFS was observed in patients with PD-L1-positive tumors when pembrolizumab was combined with chemotherapy for TNBC (overall PFS: 7.5 vs 5.6 months; PFS with composite positive score [CPS] ≥1: 7.6 vs 5.6; PFS with CPS ≥10: 9.7 vs 5.6) (Cortes J et al, KEYNOTE-355: Randomized, double-blind, phase III study of pembrolizumab + chemotherapy versus placebo + chemotherapy for previously untreated locally recurrent inoperable or metastatic triple-negative breast cancer. J Clin Oncol. 2020;38(15)). These data prompted the recent accelerated approval in the US of pembrolizumab in combination with chemotherapy for the treatment of patients with locally recurrent unresectable or metastatic TNBC (CPS ≥10) whose tumors express PD-L1, as approved by the FDA As determined by the test (KEYTRUDA ^® package insert).

Pembrolizumab (KEYTRUDA ^® ) monotherapy has demonstrated improved progression-free survival relative to chemotherapy in patients with newly diagnosed microsatellite instability-high or mismatch repair-deficient metastatic colorectal cancer. At the final analysis (median follow-up: 44.5 months [IQR 39.7-49.8]), median overall survival was not reached (NR; 95% CI 49.2-NR) with pembrolizumab and 36.7 with chemotherapy months (27.6-NR) (hazard ratio [HR] 0.74; 95% CI 0.53-1.03; p=0.036). Pembrolizumab did not show an overall survival advantage over chemotherapy because the predetermined alpha (0.025) required for statistical significance was not achieved. In this updated analysis, median progression-free survival was 16.5 months (95% CI 5.4-38.1) for pembrolizumab and 8·2 months (6.1-10.2) for chemotherapy (HR 0.59, 95% CI 0.45-0.79) (Diaz Jr. et al., "Pembrolizumab versus chemotherapy for microsatellite instability-high or mismatch repair-deficient metastatic colorectal cancer (KEYNOTE-177): final analysis of a randomised, open-label, phase 3 study”, The Lancet Oncology, Volume 23, Issue 5, P659-670, May 01, 2022. These data prompted the recent accelerated approval in the United States of pembrolizumab for the treatment of patients with Patients with locally recurrent unresectable or metastatic colon cancer of microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR) type. (KEYTRUDA ^® drug label). In non-small cell lung cancer, for example In other words, multiple randomized trials (in squamous and non-squamous histology) have demonstrated that inhibition by addition of programmed cell death protein 1 (PD-1)/programmed death ligand 1 (PD-L1) Patients with high levels of PD-L1 expression usually receive first-line pembrolizumab or atezolizumab Anti-monotherapy followed by platinum-based chemotherapy to maximize response and minimize toxicity. Those with a higher disease burden requiring more aggressive initial therapy or with lower levels of PD-L1 expression typically receive immunotherapy with Combination of platinum dual chemotherapy.

Likewise, both durvalumab and avelumab have shown promising results in the treatment of urothelial carcinoma, which currently has limited first-line chemotherapy options (see Teets et al., Avelumab: A novel anti-PD- L1 agent in the treatment of Merkel cell carcinoma and urothelial cell carcinoma. Crit Rev Immunol. 2018; 38(3): 159-206; Wills et al. Durvalumab: a newly approved checkpoint inhibitor for the treatment of urothelial carcinoma. Curr Probl C cancer . 2018; 43(3): 181-194).

Despite the compelling clinical efficacy of ICIs, up to 50% of patients with PD-L1 positive tumors display resistance or relapse after, for example, administration of ICIs (Herbst et al., Predictive correlates of response to the anti-PD-L1 Antibody MPDL3280A in cancer patients. Nature. 2014; 515(7528): 563-567). After an initial response to an immune checkpoint inhibitor, acquired resistance develops in a majority of patients (Bai et al., Regulation of PD-1/PD-L1 pathway and resistance to PD-1/PD-L1 blockade. Oncotarget . 2017; 8(66): 110693-110707). Major factors contributing to resistance include constitutive PD-L1 expression in cancer cells, tumor antigen deficiency, ineffective antigen presentation, activation of oncogenic pathways, mutations in IFN-γ signaling, and the tumor microenvironment (including exhausted T cells, Treg, bone marrow-derived Inhibitory cells (MDSC) and tumor-associated macrophages (TAM)) factors (Bai et al., Regulation of PD-1/PD-L1 pathway and resistance to PD-1/PD-L1 blockade. Oncotarget. 2017; 8 (66): 110693-110707). Other mechanisms of ICI resistance include genes that dysregulate neoantigen presentation/processing and disrupt cytotoxic T cell activity, epigenetic and cell signaling changes, and mechanisms by which noncancerous stromal or immune cells promote growth and ICI resistance (Liu et al. et al., Mechanisms of resistance to immune checkpoint blockade. Am J Clin Dermatol. 2019; 20(1): 41-54; Barrueto et al., Resistance to checkpoint inhibition in cancer immunotherapy. Transl Onc. 2020;13:100738; Jenkins et al. People, Mechanisms of resistance to immune checkpoint inhibitors. Brit J Cancer. 2018; 118: 9-16; Borcherding et al., Keeping tumors in check: A mechanistic review of clinical response and resistance to immune checkpoint blockade in cancer. J Mol Biol. 2018; 430: 2014-29; Gide et al., Primary and acquired resistance to immune checkpoint inhibitors in metastatic melanoma. Clin Cancer Res. 2018; 24(6)).

Due to the limited availability of treatments for patients with locally advanced or metastatic cancers that have progressed or are prone to develop resistance after taking immune checkpoint inhibitors, there is a need for mechanisms that broadly target these ICI resistance mechanisms and are capable of reversing or Novel therapies to prevent this resistance.

The present invention provides improved methods of treating selected patients with locally advanced, recurrent or metastatic cancer whose cancer has progressed on PD-1 or PD-L1 checkpoint inhibitors The inhibitor is resistant or susceptible to developing resistance to the inhibitor, the methods comprising administering an effective amount of a short-acting, selective and reversible cyclin-dependent kinase (CDK) 4/6 inhibitor trilaciclib or a pharmaceutically acceptable salt thereof (with a specific timed regimen) and an effective amount of a PD-1 or PD-L1 checkpoint inhibitor and an effective amount of another immune checkpoint inhibitor (ICI) selected from the group consisting of: Ig and ITIM domain T cell immune receptor (TIGIT) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint Inhibitor or cluster of differentiation 73 (CD73; exo-5' nucleotidase (NT5E)) checkpoint inhibitor.

In some aspects, an effective amount of a chemotherapeutic agent is also administered as part of a treatment regimen. In certain embodiments, administration of the selective CDK4/6 inhibitor triraciclib together with PD-1 or PD-L1 inhibitors and (importantly) other ICIs as described herein increases the potency of immune checkpoint inhibition ( Including overcoming the development of resistance to previously administered PD-1 or PD-L1 inhibitors and/or reducing or delaying the onset of resistance), thereby prolonging the efficacy of anti-cancer regimens. The therapeutic regimens described herein will reduce immune changes in the tumor microenvironment or downregulate tumor cell immune effector signals that cause tumor immune escape, and the synergistic combination of administered agents can reverse and/or significantly delay the development of ICI resistance, thereby providing Long-term efficacy of treatment regimens in hard-to-treat patient populations. In addition, the therapeutic regimens described herein provide therapeutic efficacy through immune modulation of T cells, including enhancing cytotoxic CD8+ T cell function and maturation into memory CD8+ T cells and inhibiting T _reg function and differentiation. The result of this prolonged potency is an improved host immune response to the tumor in these difficult-to-treat patients, thereby providing improved survival outcomes including overall survival (OS) and/or progression-free survival (PFS). Specific cancers amenable to treatment using the methods of the invention include, but are not limited to, non-small cell lung cancer (NSCLC), advanced/metastatic triple-negative breast cancer (TNBC), colorectal cancer, and advanced/metastatic urothelial cancer, among others.

The methods provided herein can be used to activate an immune response in a subject. For example, administration of triracicill in combination with one or more other agents can alter intratumoral immune T cell subsets to benefit effector T cell function, including stimulating IFN-γ production in exhausted T cells (Example 1, Figure 1), decrease the ratio of Treg/CD4+ T cells in the tumor (see eg Example 3, Figure 6), increase the ratio of CD8+ T cells to Treg in the tumor (see eg Example 3, Figure 7), and/or increase activation CD8+ T cells (see eg Example 3, Figure 8). Administration of triracicill in combination with an anti-PD-1 antibody resulted in a synergistic increase in IL-2 in CD4+ T cells and IFN-γ in CD8+ T cells (see eg Example 4, Figures 10A-10B). In addition, the methods provided herein can be used to reduce tumor growth and prolong the overall survival of subjects (eg, humans) with tumors. For example, administration of the CDK4/6 inhibitor treracilib in combination with a checkpoint inhibitor reduced tumor progression and prolonged overall survival compared to a regimen that did not include treracilib (see, eg, Examples 2 and 4; Figs. 2A-5B and 9). In some embodiments, the described methods provide for weekly administration of the CDK4/6 inhibitor treracilib. Weekly administration of treracilil in combination with one or more other agents provides durable tumor remission and prolongs overall survival (see, eg, Examples 2, 5, 6, 7, 8, 9, 10, and 11; Figures 2A-5B, 11A-11K, 12A-12K, 13A-13H, 15A-15Q, 16A-16B, 17A-B, 18, and 20A-20B). In contrast, delayed administration of straciclib reduced potency (eg, see Example 6, Figures 14A-14C).

Synergistic combination of the CDK4/6 inhibitor triraciclib with one or more other agents, including but not limited to, checkpoint inhibitors, can reverse and/or significantly delay resistance to checkpoint inhibition and in difficult-to-treat patient populations Provides better efficacy of treatment options. Additionally, administration of the CDK4/6 inhibitor treracilib in combination with PD-1 or PD-L1 inhibitors and other checkpoint inhibitors (e.g. selected from TIGIT inhibitors, TIM- 3 inhibitors, LAG-3 inhibitors, or checkpoint inhibitors of CD73 inhibitors) would act synergistically to substantially reduce tumor progression (see, eg, Examples 5, 6, 7, 8, 9, 10, and 11; Figure 11A- 11B, 11K, 12A-12B, 12K, 13A, 15A-15C, 15P-15Q, 16A, 17A, 18, 20A) and prolong overall survival (see for example Examples 5, 6, 7, 8, 9 and 11; Fig. 11C, 12C, 13B, 15D-15E, 16B, 17B, 20B).

The methods described herein provide enhanced anti-tumor efficacy and survival benefits in various animal cancer models including, for example, MC38 colon cancer mice (Examples 2, 3 and 4; Figures 2A-4B, 6-8 and 9-10B), CT26 colorectal cancer mice (Examples 6, 7, 8 and 11; FIGS. ), mouse mammary tumor virus-polyoma intermediate tumor antigen (MMTV-PyMT) breast cancer model mice (Example 5; Figures 11A-11K), AT3-OVA breast cancer model mice (Example 9; Figures 17A-17B), S2WTP3 Breast cancer model mice (Example 10; Figure 18). Certain cancer models, such as colorectal cancer, are often associated with the development of resistance to immune checkpoint inhibitor drugs. As shown in Figure 20A-B, in the CT-26 colorectal cancer model, the use of treracilib in combination with PD-1 or PD-L1 inhibitors and other ICIs, such as CD73 inhibitors, increased survival and decreased survival. Small tumors grow.

In some embodiments, the methods provided herein comprise administering a combination of tramacilil, a PD-1 or PD-Li inhibitor, and an additional CD73 inhibitor. CD73 is a membrane-bound extracellular enzyme that degrades adenosine triphosphate (ATP) to adenosine and is overexpressed in several types of cancer. Adenosine is an immunosuppressant that attenuates the effector function of immune cells such as T cells and enhances the suppressive function of T regs ( FIG. 19 ). Adenosine accumulation benefits tumor growth and metastasis through epidermal growth factor receptor (EGFR) signaling and inhibition of tumor cell apoptosis, while inactivation of CD73 attenuates this adenosine-mediated process. CD73 also releases matrix metalloproteinases (MMPs) that facilitate the breakdown of extracellular matrix (ECM), thereby enabling tumor cells to invade and migrate to distant organs.

Mechanisms of ICI resistance include genetic, epigenetic and cell signaling alterations that dysregulate neoantigen presentation/processing and disrupt cytotoxic T cell activity, and mechanisms by which noncancerous stromal or immune cells promote growth and ICI resistance. Another mechanism is other co-stimulatory checkpoints such as T cell immune receptor with immunoglobulin and ITIM domains (TIGIT), T cell immunoglobulin and mucin domain 3 (TIM-3), lymphocyte activation gene -3 (LAG-3) and cluster of differentiation 73 (CD73; exo-5' nucleotidase (NT5E)) checkpoint). By administering triracicil with PD-1 or PD-L1 inhibitors and other costimulatory immune checkpoint inhibitors selected from TIGIT inhibitors, LAG-3 inhibitors, TIM-3 inhibitors or CD73 inhibitors and visual In the case of combinations of chemotherapeutic agents as described herein, one or more of the mechanisms responsible for ICI resistance and disease progression can be overcome, thereby increasing antigen presentation (major histocompatibility complex (MHC) class I), enhancing T cell Selective and tumor infiltration, inhibition of regulatory T cell proliferation, reduction of markers of T cell exhaustion (programmed cell death protein 1 (PD-1), cytotoxic T lymphocyte-associated protein 4 (CTLA‑4), T cell immunity Globulin and mucin domain 3 (TIM3)) expression, stabilize PD-L1 expression on tumor cells, promote dendritic cell migration, adenosine or increase T effector through high interferon gamma (IFN-γ) production cell function. By administering the therapeutic combinations described herein to this difficult-to-treat subgroup of patients, the immunosuppressive tumor microenvironment in the patient's tumor that renders previously administered ICIs ineffective or less effective and allows tumor progression can be enhanced Significantly overcome, thereby improving the ability of the patient's immune system to reduce or control tumor burden, improve quality of life, and improve overall survival in this difficult-to-treat subgroup of patients.

In some aspects, the treatment regimen is administered to patients with advanced or metastatic cancer whose disease has progressed (indicative of immune checkpoint inhibitor resistance) following prior treatment with a PD-1 or PD-L1 inhibitor. Therefore, these patients are administered treacilil, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors, wherein other checkpoint inhibitors are selected from TIGIT inhibitors, TIM-3 inhibitors, LAG- 3 inhibitors or CD73 inhibitors. In some embodiments, straciclib, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors are administered on Day 1 of each 14-day therapeutic treatment cycle (or cycles), and at Trascilli was administered again on Day 7 of each 14-day cycle. In some embodiments, straciclib, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors are administered on Day 1 of each 21-day cycle, and administered on Days 7 and 10 of each 21-day cycle. On the 14th day, Traxil was administered again. In some embodiments, treracilib is administered one or more times per week. In some embodiments, treracilil, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors are administered on Day 1 of every 28-day cycle, and administered on Days 7, 10 of each 21-day cycle. On the 14th day and the 21st day, Trirascillin was administered again. In some embodiments, treracilib is administered one or more times per week. In some embodiments, treracilil, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors are administered on Day 1 of each 42-day cycle, and on Day 7 of each 42-day cycle , On the 14th, 21st, 28th and 35th days, treracilib was administered again.

In some embodiments, treracilib is administered one or more times per week. In some embodiments, treracillib is administered once a week during the treatment period, and is administered once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, or once every six weeks PD-1 or PD-L1 and other immune checkpoint inhibitors. In some embodiments, treracilib is administered one or more times per week. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 inhibitor is administered according to its standard label for administration for its approved use, and the other immune checkpoint inhibitor is administered with PD-L1 or PD-1 inhibitors were administered concurrently. In some embodiments, other immune checkpoint inhibitors are not administered concomitantly with the PD-L1 or PD-1 inhibitor. In some embodiments, treracilib is administered one or more times per week. In some embodiments, only the initial loading dose of treracilil is administered about 8 days, 7 days, 6 days, 5 days, 4 days, or 3 days prior to beginning the first treatment cycle as set forth above.

In some embodiments, the cancer is non-small cell lung cancer, triple negative breast cancer, colorectal cancer, or urothelial cancer. In some embodiments, the patient has second-line metastatic non-squamous or squamous NSCLC. In some embodiments, the patient has second-line locally advanced or metastatic urothelial carcinoma. In some embodiments, the colorectal cancer is confirmed by laboratory testing to be of the microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR) type.

In some alternative embodiments, the other immune checkpoint inhibitor administered in combination with treracicill and a PD-1 or PD-L1 inhibitor is selected from the group consisting of programmed death-ligand 2 (PD-L2) inhibitors, CTLA- 4 inhibitors and V domain Ig inhibitors of T cell activation (VISTA) inhibitors, B7-H3/CD276 inhibitors, indoleamine 2,3-dioxygenase (IDO) inhibitors, killer immunoglobulin-like Receptor (KIR) inhibitors, carcinoembryonic antigen cell adhesion molecule (CEACAM) (eg, CEACAM-1, CEACAM-3, and CEACAM-5) inhibitors, sialic acid-binding immunoglobulin-like lectin 15 (Siglec-15) inhibitors inhibitors, CD47 inhibitors, CD39 inhibitors, or B and T lymphocyte attenuating protein (BTLA) inhibitors.

In an alternative to the above embodiment, the treatment regimen administers treracilil in combination with PD-1 or PD-L1 as described herein and various tyrosine kinase (MTK) inhibitors (in place of other immune checkpoint inhibitors) , such as, but not limited to, lenvatinib, sitravatinib, and cabozantinib).

In yet another alternative to the above embodiments, the treatment regimen administers treracilil and PD-1 or PD-L1 as described herein and an oncolytic virus (such as, but not limited to, oncolytic adenovirus, oncolytic adenovirus, oncolytic HSV-1, oncolytic reovirus, oncolytic poxvirus, oncolytic Newcastle disease virus, oncolytic measles virus, oncolytic Seneca Valley virus, Japanese hemagglutination viral envelope (HVJ-E) virus, oncolytic gamma-herpes virus, oncolytic parvovirus, or oncolytic retrovirus). Oncolytic viruses include (but are not limited to) paleorep (Reolysin ^® , Oncolytics Biotech), Japanese hemagglutination virus envelope (GEN0101), seprehvir, talimogene laherparepvec , T-VEC), adenovirus VCN-01, adenovirus ICORVIR-5, HF10, GL-ONC1, DNX-2401 and enadenotucirev.

In some embodiments, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15 , at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24 or more treatment cycles. In some embodiments, the patient is administered treatment cycles until disease progression.

In some aspects, the patient is administered triraciclib, a chemotherapeutic agent, a PD-1 or PD-L1 inhibitor, other immune checkpoint inhibitor, wherein the other checkpoint inhibitor is selected from TIGIT inhibitors, TIM- 3 inhibitors, LAG-3 inhibitors or CD73 inhibitors and other cytotoxic chemotherapeutic agents, wherein before administration of chemotherapeutic agents (for example, less than 24 hours, less than 16 hours, less than 12 hours before each administration of chemotherapeutic agents) hours, less than 8 hours, less than 6 hours, less than 4 hours, less than 2 hours, less than 1 hour, or about 30 minutes), and on Day 1 of each therapeutic treatment cycle (or cycles) Administer PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors.

It has been found that by using triaciclib during a chemotherapeutic/immune checkpoint inhibitor combination regimen, immune effector cells (such as T lymphocytes) are protected from chemotherapeutic agent toxicity and in chemotherapy-induced immunogenic cells Release from transient cell cycle arrest in the presence of death significantly improved the initiation and activation of anticancer immune responses and anticancer effects compared to the absence of treracilib. It has also been found that by using treracicill during a chemotherapeutic/immune checkpoint inhibitor regimen, antitumor activity is enhanced by cell cycle independent and dependent mechanisms involving selective reduction of intratumoral T _reg populations , Protect pro-inflammatory immune effector cells (such as tumor infiltrating lymphocytes) and increase the durability of therapeutic responses. Compared with continuous inhibition of CDK4/6 with chemotherapy and immune checkpoint inhibitors alone or with daily CDK4/6 inhibitors (including combinations of long-acting CDK4/6 inhibitors and immune checkpoint inhibitors), Controlled inhibition of CDK4/6 using the combination of treracilil with chemotherapeutics and various immune checkpoint inhibitors as described herein can significantly increase the anti-tumor effect. The chemotherapeutic agent to be administered may be one that is typically administered as part of the standard of care for the stage of cancer being treated. Many chemotherapeutics (such as, but not limited to, protein synthesis inhibitory or DNA-damaging chemotherapeutics) tend to be nonspecific and toxic to rapidly dividing normal cells, including immune effector cells, and hematotoxic (such as myelosuppressive) It is a common side effect of chemotherapy. Proliferation of immune effector cells generally requires CDK4/6 activity, i.e., it is CDK4/6 replication dependent (see Roberts et al., Multiple Roles of Cyclin-Dependent Kinase 4/6 Inhibitors in Cancer Therapy. JNCI 2012; 104(6 ):476-487). All major intratumoral immune cell types such as CD4+ T cells, CD8+ T cells and natural killer (NK) cells are sensitive to CDK4/6 inhibition. Immune effector cells, which are sensitive to the damaging effects of chemotherapeutic agents during proliferation, are temporarily arrested in the G0/G1 phase of the cell cycle during proliferation by the use of treracilil during chemotherapy treatment. By protecting these cells from the damaging effects of chemotherapeutic agents, administration of specifically timed treracilil protects immune function, enhances T cell activation, and increases potency of immune checkpoint inhibitors, resulting in significantly improved anticancer immune response. Importantly, the development of resistance to the effects of immune checkpoint inhibitors (due to long-term use and immune changes in the tumor microenvironment or tumor cell immune effects) can be reversed or significantly delayed by including other immune checkpoint inhibitors in the treatment regimen. signal downregulation), thereby providing long-term efficacy of treatment regimens in difficult-to-treat patient populations.

It has also been found that certain chemotherapy treatments, but not all, are capable of triggering a pathway called "immunogenic cell death" ("ICD") in tumor cells (see generally Locy, H. et al., Immunomodulation of the Tumor Microenvironment: Turn Foe Into Friend, Frontiers in Immunology, 2018; 9: 2090). The ICD is a form of regulated cell death that induces the release of tumor-associated antigens and triggers an anti-tumor immune response (ibid.). ICDs involve the release of damage-associated molecular pathways ("DAMPs") that alert host immune system cells to damage. There are six cell death-promoting DAMPs: Calreticulin ("CRT"), High Mobility Group Box 1 (HMGB1), Extracellular ATP, Type I Interferon, Cancer Cell-Derived Nucleic Acid, and ANXA1. These DAMPs determine the strength and durability of the ICD's antitumor response. See also Wang et al., Immunogenic effects of chemotherapy induced tumor cell death, Genes & Diseases (2018) 5, 194-203.

Chemotherapeutics can also induce immunogenic effects by destroying strategies used by tumors to evade immune responses. See, eg, Emens et al., The Interplay of Immunotherapy and Chemotherapy: Harnessing Potential Synergies. Cancer Immunol Res; 3(5), May 2015. For example, chemotherapy can modulate different features of tumor immunobiology in a drug-, dose-, and schedule-dependent manner, and different chemotherapeutic drugs can modulate intrinsic immunogenicity of tumor cells through different mechanisms (see, for example, Chen G, Emens LA. Chemoimmunotherapy: reengineering tumor immunity. Cancer Immunol Immunother 2013; 62: 203-16.). Chemotherapy can also enhance tumor antigen presentation by upregulating the expression of tumor antigens themselves or the MHC class I molecules to which these antigens bind. Alternatively, chemotherapy can upregulate co-stimulatory molecules (B7-1) or down-regulate co-inhibitory molecules (PD-L1/B7-H1 or B7-H4) expressed on the surface of tumor cells, thereby enhancing the intensity of effector T cell activity. Chemotherapy can also render tumor cells more sensitive to T cell-mediated lysis through fas-, perforin- and granzyme B-dependent mechanisms.

PD1 inhibitors for use in the methods described herein include, for example but not limited to, nivolumab ( ^OPDIVO® ; Bristol Myers Squibb), pembrolizumab ( ^KEYTRUDA® ; Merck), cimipril Anti (LIBTAYO ^® ; Regeneron), dostarlimab (JEMPERLI ^® ; GlaxoSmithKline), pidilizumab (Medivation), AMP-224 (AstraZeneca/Medimmune), AMP-514 (AstraZeneca) , sintilimab (IBI308; Innovent/Eli Lilly), sasanlimab (PF-06801591; Pfizer), spartalizumab (PDR001; Novartis), Reveli Retifanlimab (MGA012/INCMGA00012; Incyte Corporation and MacroGenics), tislelizumab (BGB-A317; BeiGene), toripalimab (JS001; Coherus BioSciences, Inc. and Shanghai Junshi Biosciences Co., Ltd.), camrelizumab (camrelizumab) (SHR-1210; Jiangsu Hengrui Medicine Company and Incyte Corporation), CS1003 (Cstone Pharmaceuticals), zimberelimab (AB122 ; Arcus Biosciences) and JTX-4014 (Jounce Therapeutics).

PD-L1 inhibitors for use in the methods described herein include, for example but not limited to, atezolizumab (TECENTRIQ ^® , Genentech), durvalumab (IMFINZI ^® , AstraZeneca), avelumab (BAVENCIO ^® ; Merck), envafolimab (KN035; Alphamab), BMS-936559 (Bristol-Myers Squibb), BMS-986189 (Bristol-Myers Squibb), lodapolimab (LY3300054 ; Eli Lilly), cosibelimab (CK-301; Checkpoint Therapeutics), sugemalimab (CS-1001; Cstone Pharmaceuticals), adebrelimab (SHR-1316 ; Jiangsu HengRui Medicine), CBT-502 (CBT Pharma), AUNP12 (Aurigene), CA-170 (Aurigene/Curis) and BGB-A333 (BeiGene). In some embodiments, a dual PD-L1/PD-1 inhibitor as provided herein is administered.

LAG-3 inhibitors for use in the methods described herein include, for example but not limited to, relatlimab ( ^OPDUALAG® ; BMS-986016; Bristol-Myers Squibb), GSK2831781 (GlaxoSmithKline), Effie eftilagimod alpha (IMP321; Prima BioMed), leramilimab (LAG525; Novartis), favezelimab (MK-4280; Merck), Anti-(fianlimab) (REGN3767; Regeneron), TSR-033 (Tesaro/GSK), BI754111 (Boehringer Ingelheim), Sym022 (Symphogen), LBL-007 (Nanjing Leads Biolabs Co., Ltd), IBI110 (Innovent Biologics), IBI323 (Innovent Biologics), INCAGN02385 (Incyte Corporation), MGD013 (Macrogenics), RO7247669 (Hoffman-LaRoche), EMB-02 (Shanghai Epimab Biotherapeutics), XmAb841 (Xencor), dual PD-1 and LAG-3 inhibitor Tepoli Tebotelimab (MGD013; MacroGenics), CB213 (Crescendo Biologics) and SNA-03 (Microbio Group) and the dual PD-L1 and LAG-3 inhibitor FS118 (F-Star).

TIM-3 inhibitors for use in the methods described herein include, for example but not limited to, Cobolimab (TSR-022; Tesaro), RG7769 (Genentech), MAS825 (Novartis), Sabato sabatolimab (MBG453; Novartis), Sym023 (Symphogen), INCAGN2390 (Incyte), LY3321367 (Eli Lilly and Company), BMS-986258 (BMS), SHR-1702 (Jiangsu HengRui), AZD7789 (AstraZeneca), TQB2618 (Chia Tai Tianqing Pharmaceutical Group Co., Ltd.) and NB002 (Neologics Bioscience), BGBA425 (Beigene) and Tim-3 and PD-1 bispecific RO7121661 (Roche).

TIGIT (T cell immune receptor with Ig and ITIM domains) inhibitors for use in the methods described herein include, for example but not limited to, Vibostolimab (MK-7684; Merck), Etivir Etigilimab/OMP-313 M32 (OncoMed), Tiragolumab (MTIG7192A/RG-6058; Roche/Genentech), Ociperlimab (BGB-A1217; Beigene), BMS-986207 (BMS), COM902 (Compugen), M6223 (Merck KGaA), domvanalimab (AB-154; Arcus Biosciences), AZD2936 (AstraZeneca), JS006 (Shanghai Junshi Biosciences), IBI139 (Innovent Biologics), ASP-8374 (Astellas/Potenza), BAT6021 (Bio-Thera Solutions), TAB006 (Shanghai Junshi Bioscience), Domvanalimab (AB154; Arcus Biosciences), EOS884448 (EOS-448; iTeos), SEA-TGT (Seattle Genetics), mAb-7 (Stanwei Biotech), SHR-1708 (Hengrui Medicine), GS02 (Suzhou Zelgen/Qilu Pharma), RXI-804 (Rxi Pharmaceuticals), NB6253 (Northern Biologics), ENUM009 (Enumreal Biomedical), CASC-674 (Cascadian Therapeutics), AJUD008 (AJUD Biopharma), AGEN1777 (Agenus, Bristol-Myers Squibb), anti-TIGIT/anti-PD-L1 bispecific antibody HLX301 (Shanghai Henlius Biotech), HLX53 (Shanghai Henlius Biotech), BAT6005 (Bio-Thera Solutions) and anti-TIGIT/anti-PD-L1 antibody HB0036 (Shanghai Huaota Biopharmaceutical).

Cluster of differentiation (CD) 73 (exo-5' nucleotidase; (NT5E) checkpoint inhibitors for use in the methods described herein include, for example but not limited to, HLX23 (Shanghai Henlius Biotech), LY3475070 (Eli Lilly and Company), IPH5301 (Innate Pharma, Astra Zeneca), AK119 (Akesobio Australia Pty Ltd.), PT199 (Phanes Therapeutics), mupadolimab (CPI-006; Corvus Pharmaceuticals), Sym024 (Symphogen), Oleclumab (MEDI9447; Astra Zeneca), IBI325 (Innovent Biologics), ORIC-533 (Oric Pharmaceuticals), JAB-BX102 (Jacobio Pharmaceuticals), TJ004309 (Tracon Pharmaceuticals), AB680 (Arcus Biosciences), NZV9 30 (Novartis), BMS-986179 (Bristol Myers Squibb), INCA00186 (Incyte Corporation) and anti-CD73-TGFβ-Trap bifunctional antibody dalutrafusp alfa (Gilead Sciences).

Chemotherapeutic agents for use in the methods of the invention include those associated with standard-of-care treatments for the particular stage and prior treatment state of the cancer being treated. Examples of suitable chemotherapeutic agents for use in the methods of the invention include, but are not limited to: platinum-containing drugs such as carboplatin, cisplatin, and oxaliplatin; taxanes; ), such as paclitaxel, docetaxel, or albumin-stabilized nanoparticle formulations of paclitaxel (nab-paclitaxel); topoisomerase inhibitors, such as topotecan ), campothecin, irinotecan, belotecan, doxorubicin, daunorubicin, epirubicin, idarubicin idarubicin, etoposide, and teniposide; cyclophosphamide; vinblastine; gemcitabine; 5-fluorouracil (5-FU); eribulin; pemetrexed; mitomycin; sacituzumab govitecan; valrubicin; vinorelbine tartrate; and pharmaceutically acceptable salts thereof; or a combination thereof. In some embodiments, the chemotherapeutic agent used in the methods of the invention is a chemotherapeutic agent capable of inducing an immune-mediated response. Chemotherapies capable of inducing immune-mediated responses are generally known in the art and include, but are not limited to, alkylating agents such as cyclophosphamide, trabectedin, temozolomide, melphalan (melphalan), dacarbazine, and oxaliplatin; antimetabolites such as methotrexate, mitoxantrone, gemcitabine, and 5-fluorouracil (5-FU); cytotoxicity Antibiotics, such as bleomycin and anthracyclines (including doxorubicin, daunorubicin, epirubicin, idarubicin, and valrubicin); taxanes, such as paclitaxel, cabazitaxel cabazitaxel and docetaxel; topoisomerase inhibitors such as topotecan, irinotecan and etoposide; platinum compounds such as carboplatin, oxaliplatin and cisplatin; bortezomib ( bortezomib), 26S proteasome subunit inhibitors; vinca alkaloids such as vinblastine, vincristine, vindesine, and vinorelbine; diaziquone; mechlorethamine; mitomycin C; fludarabine; cytosine arabinoside; or a pharmaceutically acceptable salt thereof and any combination thereof. In some embodiments, the chemotherapy is selected from idarubicin, epirubicin, doxorubicin, mitoxantrone, oxaliplatin, bortezomib, gemcitabine, and cyclophosphamide, or any pharmaceutical Acceptable salts and any combination thereof.

In some embodiments, treracicill is administered daily during the cycle of chemotherapeutic administration, a PD-1 or PD-L1 inhibitor is administered at least on day 1 of each cycle, and the combination of PD-1 or PD - the L1 inhibitor is administered concomitantly with another immune checkpoint inhibitor selected from a TIGIT inhibitor, a TIM-3 inhibitor, a LAG-3 inhibitor, or a CD73 inhibitor. In some embodiments, the PD-1 or PD-L1 inhibitor is not administered concurrently with other checkpoint inhibitors. In some embodiments, the PD-1 inhibitor or PD-L1 inhibitor and other immune checkpoint inhibitors are administered every two weeks. In some embodiments, the PD-1 inhibitor or PD-L1 inhibitor and other immune checkpoint inhibitors are administered every three weeks. In some embodiments, the PD-1 inhibitor or PD-L1 inhibitor and other immune checkpoint inhibitors are administered every four weeks. In some embodiments, the PD-1 inhibitor or PD-L1 inhibitor and other immune checkpoint inhibitors are administered every six weeks. In some embodiments, the PD-1 inhibitor or PD-L1 inhibitor is administered once for the duration of the first cycle and the other immune checkpoint inhibitor is administered once for the duration of the second cycle, wherein the first The duration of the cycle is selected from two weeks, three weeks, four weeks or six weeks, wherein the duration of the second cycle is selected from two weeks, three weeks, four weeks or six weeks, and wherein the duration of the first cycle is the same as the duration of the second cycle The duration of the cycle varies. In some embodiments, after cycle day 1, treracillib is further administered alone once a week during the cycle. For example, in some embodiments, treracilil, chemotherapeutic agents, PD-1 or PD-L1 inhibitors, and other checkpoint inhibitors are administered on Day 1 of a 14-day cycle. In some embodiments, treracicill is re-administered on day 7 of each 14-day cycle. In some embodiments, triraciclib and a chemotherapeutic agent are administered on Day 1 of a 21-day cycle, and a PD-1 or PD-L1 inhibitor and other immune checkpoint inhibition are administered on Day 1 of the 21-day cycle agent. In some embodiments, treracicill is further administered alone on days 7 and 14 of the 21-day cycle. In some embodiments, triraciclib and chemotherapeutics are administered on days 1-3 of a 21-day cycle, and a PD-1 or PD-L1 inhibitor and other immune tests are administered on day 1 of the 21-day cycle point inhibitors. In some embodiments, treracicill is further administered alone on days 7 and 14 of the 21-day cycle. In some embodiments, treracillib and the chemotherapeutic agent are administered on days 1, 8, and 15 of a 28-day cycle, and a PD-1 or PD-L1 inhibitor is administered on day 1 of the 28-day cycle along with other agents. Immune checkpoint inhibitors. In some embodiments, treracilil, chemotherapeutics, PD-1 or PD-L1 inhibitors, and other immune checkpoint inhibitors are administered on Day 1 of every 28-day cycle. In some embodiments, treracicill is re-administered on days 7, 14, and 21 of each 28-day cycle. In some embodiments, treracilib is administered one or more times per week. In some embodiments, the patient is administered 2 or more treatment cycles, eg, at least 2 cycles, at least 3 cycles, at least 4 cycles, at least 5 cycles, at least 6 cycles or more. In some embodiments, following discontinuation of a treatment cycle that includes a chemotherapeutic agent, the patient is administered treaciclib, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors in one or more maintenance phase cycles, Among them, once a week, once every two weeks, once every three weeks, once every four weeks, or once every six weeks, treracilil, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors were simultaneously administered. In some embodiments, the PD-1 inhibitor or PD-L1 inhibitor and other immune checkpoint inhibitors are administered every two weeks. In some embodiments, the PD-1 inhibitor or PD-L1 inhibitor and other immune checkpoint inhibitors are administered every three weeks. In some embodiments, the PD-1 inhibitor or PD-L1 inhibitor and other immune checkpoint inhibitors are administered every four weeks. In some embodiments, the PD-1 inhibitor or PD-L1 inhibitor and other immune checkpoint inhibitors are administered every six weeks. In some embodiments, the PD-1 or PD-L1 inhibitor is not administered concurrently with other checkpoint inhibitors. In some embodiments, the PD-1 inhibitor or PD-L1 inhibitor is administered once for the duration of the first cycle and the other immune checkpoint inhibitor is administered once for the duration of the second cycle, wherein the first The duration of the cycle is selected from two weeks, three weeks, four weeks or six weeks, wherein the duration of the second cycle is selected from two weeks, three weeks, four weeks or six weeks, and wherein the duration of the first cycle is the same as the duration of the second cycle The duration of the cycle varies. In some embodiments, following discontinuation of a treatment cycle that includes a chemotherapeutic agent, the patient is administered treaciclib, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors in one or more maintenance phase cycles, Among them, triraciclib is administered once a week and PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors are administered once a week, once every two weeks, once every three weeks, once every four weeks, or once every six weeks agent. In some embodiments, following discontinuation of a treatment cycle that includes a chemotherapeutic agent, the patient is administered treaciclib, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors in one or more maintenance phase cycles, wherein triracicill is administered weekly, wherein a PD-1 or PD-L1 inhibitor is administered once for the duration of the first cycle, and wherein the other immune checkpoint inhibitor is administered once for the duration of the second cycle , where the duration of the first period differs from the duration of the second period.

Non-Small Cell Lung Cancer (NSCLC) In one aspect, the treatment regimens described herein are administered to patients with metastatic or locally advanced non-small cell lung cancer (NSCLC). In some embodiments, the patient has metastatic or locally advanced squamous cell NSCLC. In some embodiments, the patient has metastatic or locally advanced non-squamous NSCLC. In some embodiments, the patient is not eligible for therapy targeting a driver NSCLC mutation. In some embodiments, the patient has NSCLC with a driver mutation, but the patient is not a candidate for therapy targeting the driver NSCLC mutation. In some embodiments, the patient has NSCLC with unknown driver mutation status.

Squamous NSCLC In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic squamous NSCLC in a first-line advanced/metastatic setting, wherein the patient is administered Qu Rascilli, one or more chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1, as determined by a USFDA approval test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 (as determined by a Tumor Proportion Score (TPS)), where TPS is the number of PD-L1 positive tumor cells divided by PD-L1 positive + PD-L1 negative The total number of tumor cells is multiplied by 100 as determined by FDA approval test or CE mark test. In some embodiments, TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 as determined by staining immune cells (%IC). In some embodiments, the IC > 10%. In some embodiments, the patient has a tumor expressing PD-L1, as determined by staining tumor cells (%TC). In some embodiments, TC > 1%. In some embodiments, TC > 25%. In some embodiments, TC > 50%. In some embodiments, the chemotherapeutic agent administered is a standard-of-care chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is a combination of chemotherapeutic agents selected from gemcitabine, vinorelbine, paclitaxel, nab-paclitaxel, and a platinum drug such as cisplatin, carboplatin, or oxaliplatin. In some embodiments, the chemotherapeutic agent is gemcitabine and carboplatin or cisplatin. In some embodiments, the chemotherapeutic agents are paclitaxel and carboplatin. In some embodiments, the chemotherapeutic agent is nab-paclitaxel and carboplatin. In some embodiments, the chemotherapeutic agent is docetaxel. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic squamous cell NSCLC in a second-line advanced/metastatic setting, wherein the patient is administered triaciclib , one or more chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 1%. In some embodiments, the chemotherapeutic agent administered is a standard-of-care chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is selected from docetaxel, gemcitabine, vinorelbine, or combinations thereof. In some embodiments, the chemotherapeutic agent is docetaxel. In some embodiments, the chemotherapeutic agent is gemcitabine. In some embodiments, the chemotherapeutic agent is vinorelbine. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic squamous cell NSCLC in a second-line advanced/metastatic setting, wherein the patient is administered triaciclib , PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 1%. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic squamous cell NSCLC in a second-line advanced/metastatic setting, wherein the patient is administered triaciclib , PD-1 or PD-L1 inhibitors and multiple tyrosine kinase (MTK) inhibitors. In some embodiments, the MTK inhibitor is selected from levatinib, stratinib or crizotinib. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 1%.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic squamous cell NSCLC in a first-line or second-line advanced/metastatic setting, wherein the patient is administered tretinoin Rascilli, PD-1 or PD-L1 inhibitors, and oncolytic viruses. In some embodiments, the oncolytic virus is selected from the group consisting of oncolytic adenovirus, oncolytic HSV-1, oncolytic Leo virus, oncolytic pox virus, oncolytic Newcastle disease virus, oncolytic measles virus, oncolytic Seneca Valley virus, Japanese hemagglutinating virus envelope (HVJ-E) virus, oncolytic gamma herpes virus, oncolytic parvovirus or oncolytic retrovirus. In some embodiments, the oncolytic virus is Palerip. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 10%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 25%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 50%. In some embodiments, the patient is also administered a drug selected from pemetrexed, gemcitabine, paclitaxel, nab-paclitaxel, and a platinum drug (eg, cisplatin, carboplatin, or oxaliplatin), docetaxel, or vincin Chemotherapeutic agent of Ruibine or any pharmaceutically acceptable salt or combination thereof.

Non-squamous NSCLC In one aspect, the improved method of treatment described herein is administered to patients with locally advanced or metastatic non-squamous NSCLC in a first-line advanced/metastatic setting, wherein the patient is administered with triracicill, one or more chemotherapeutic agents, PD-1 or PD-L1 inhibitors, and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors, or CD73 inhibitors . In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 10%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 25%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 50%. In some embodiments, the chemotherapeutic agent administered is a standard-of-care chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is a combination of chemotherapeutic agents selected from pemetrexed, gemcitabine, paclitaxel, nab-paclitaxel, and a platinum drug such as cisplatin, carboplatin, or oxaliplatin. In some embodiments, the chemotherapeutic agent is gemcitabine and carboplatin or cisplatin. In some embodiments, the chemotherapeutic agents are paclitaxel and carboplatin. In some embodiments, the chemotherapeutic agent is nab-paclitaxel and carboplatin. In some embodiments, the chemotherapeutic agent is docetaxel. In some embodiments, the chemotherapeutic agent is pemetrexed and carboplatin, cisplatin, or oxaliplatin. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic non-squamous NSCLC in a second-line advanced/metastatic setting, wherein the patient is administered trela Sealy, one or more chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 1%. In some embodiments, the chemotherapeutic agent administered is a standard-of-care chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is selected from docetaxel, gemcitabine, vinorelbine, or combinations thereof. In some embodiments, the chemotherapeutic agent is docetaxel. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic non-squamous NSCLC in a second-line advanced/metastatic setting, wherein the patient is administered trela cilium, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 1%. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic non-squamous NSCLC in a second-line advanced/metastatic setting, wherein the patient is administered trela Sealy, PD-1 or PD-L1 inhibitors and multiple tyrosine kinase (MTK) inhibitors. In some embodiments, the MTK inhibitor is selected from levatinib, stretratinib, or crizotinib. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 1%.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic non-squamous NSCLC in a first-line or second-line advanced/metastatic setting, wherein the patient is administered Triracicill, PD-1 or PD-L1 inhibitors, and oncolytic viruses. In some embodiments, the oncolytic virus is selected from the group consisting of oncolytic adenovirus, oncolytic HSV-1, oncolytic Leo virus, oncolytic pox virus, oncolytic Newcastle disease virus, oncolytic measles virus, oncolytic Seneca Valley virus, Japanese hemagglutinating virus envelope (HVJ-E) virus, oncolytic gamma herpes virus, oncolytic parvovirus or oncolytic retrovirus. In some embodiments, the oncolytic virus is Palerip. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 10%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 25%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 50%. In some embodiments, the patient is also administered a drug selected from pemetrexed, gemcitabine, paclitaxel, nab-paclitaxel, and a platinum drug (eg, cisplatin, carboplatin, or oxaliplatin), docetaxel, or vincin Chemotherapeutic agent of Ruibine or any pharmaceutically acceptable salt or combination thereof.

Triple Negative Breast Cancer In one aspect, the modified treatment approach described herein is administered to patients with locally advanced, recurrent unresectable or metastatic triple negative breast cancer (TNBC) in a first-line advanced/metastatic setting and A patient whose tumor expresses PD-L1, and wherein the patient is administered triraciclib, one or more chemotherapeutic agents, a PD-1 or PD-L1 inhibitor and a TIGIT inhibitor, a TIM-3 inhibitor, a LAG- 3 inhibitors or other immune checkpoint inhibitors of CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1 and a composite positive score (CPS) > 10. In some embodiments, the chemotherapeutic agent administered is a standard-of-care chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is selected from gemcitabine, vinorelbine, capecitabine, ixabepilone, eribulin, paclitaxel, nab-paclitaxel, cyclophosphine Chemotherapeutic agents of amines, 5-fluorouracil, doxorubicin, gosatuzumab, etoposide or platinum drugs such as cisplatin, carboplatin or oxaliplatin, or any pharmaceutically acceptable salt thereof or a combination thereof. In some embodiments, the chemotherapeutic agent is gemcitabine and carboplatin or cisplatin. In some embodiments, the chemotherapeutic agent is paclitaxel or nab paclitaxel and carboplatin. In some embodiments, the chemotherapeutic agents are doxorubicin and cyclophosphamide. In some embodiments, the chemotherapeutic agents are capecitabine, docetaxel, and cyclophosphamide. In some embodiments, the chemotherapeutic agent is docetaxel, cyclophosphamide and paclitaxel or nab-paclitaxel. In some embodiments, the chemotherapeutic agents are capecitabine and cisplatin. In some embodiments, the chemotherapeutic agents are docetaxel and capecitabine. In some embodiments, the chemotherapeutic agents are ixabepilone and capecitabine. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved methods of treatment described herein are administered to patients with locally advanced, recurrent unresectable or metastatic triple-negative breast cancer (TNBC) and their tumors in a second-line advanced/metastatic setting A patient exhibiting PD-L1, and wherein the patient is administered treracilil, one or more chemotherapeutic agents, a PD-1 or PD-L1 inhibitor and a TIGIT inhibitor, a TIM-3 inhibitor, a LAG-3 inhibitor other immune checkpoint inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1 and a composite positive score (CPS) > 10. In some embodiments, the chemotherapeutic agent administered is a standard-of-care chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is selected from gemcitabine, vinorelbine, capecitabine, ixabepilone, eribulin, paclitaxel, nab-paclitaxel, cyclophosphamide, 5-fluorouracil, Chemotherapeutic agents of doxorubicin, gosatuzumab, etoposide or platinum drugs (such as cisplatin, carboplatin or oxaliplatin) or any pharmaceutically acceptable salts or combinations thereof. In some embodiments, the chemotherapeutic agent is gemcitabine and carboplatin or cisplatin. In some embodiments, the chemotherapeutic agent is paclitaxel or nab paclitaxel and carboplatin. In some embodiments, the chemotherapeutic agents are doxorubicin and cyclophosphamide. In some embodiments, the chemotherapeutic agents are capecitabine, docetaxel, and cyclophosphamide. In some embodiments, the chemotherapeutic agent is docetaxel, cyclophosphamide and paclitaxel or nab-paclitaxel. In some embodiments, the chemotherapeutic agents are capecitabine and cisplatin. In some embodiments, the chemotherapeutic agents are docetaxel and capecitabine. In some embodiments, the chemotherapeutic agents are ixabepilone and capecitabine. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic TNBC in a second-line advanced/metastatic setting, wherein the patient is administered triraciclib, PD- 1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1 and a CPS > 10. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic TNBC in a second-line advanced/metastatic setting, wherein the patient is administered triraciclib, PD- 1 or PD-L1 inhibitors and oncolytic viruses. In some embodiments, the oncolytic virus is selected from the group consisting of oncolytic adenovirus, oncolytic HSV-1, oncolytic Leo virus, oncolytic pox virus, oncolytic Newcastle disease virus, oncolytic measles virus, oncolytic Seneca Valley virus, Japanese hemagglutinating virus envelope (HVJ-E) virus, oncolytic gamma herpes virus, oncolytic parvovirus or oncolytic retrovirus. In some embodiments, the oncolytic virus is Palerip. In some embodiments, the patient has a tumor expressing PD-L1. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a PD-L1 expressing tumor with a composite positivity score (CPS) >10. In some embodiments, the patient is also administered a drug selected from gemcitabine, vinorelbine, capecitabine, ixabepilone, eribulin, paclitaxel, nab-paclitaxel, cyclophosphamide, 5-fluorouracil , doxorubicin, gosatuzumab, etoposide or a chemotherapeutic agent of a platinum drug (such as cisplatin, carboplatin or oxaliplatin) or any pharmaceutically acceptable salt or combination thereof.

Colorectal Cancer In one aspect, the improved method of treatment described herein is administered to patients with locally advanced or metastatic and unresectable colorectal cancer in the first-line advanced/metastatic setting, wherein the patient is given Administration of triracicill, one or more chemotherapeutic agents, PD-1 or PD-L1 inhibitors, and other immune checkpoint inhibition selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors, or CD73 inhibitors agent. In some embodiments, the patient has a tumor that is microsatellite instability-high (MSI-H) or mismatch repair-deficient (dMMR)-type as confirmed by an FDA-approved test. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 (as determined by the composite positive score (CPS)), which is the number of PD-L1 staining cells (tumor cells, lymphocytes, macrophages) divided by The total number of viable tumor cells was multiplied by 100, as determined by an FDA-approved test. In some embodiments, the tumor has a CPS > 10. In some embodiments, the patient has a tumor expressing PD-L1 (as determined by a Tumor Proportion Score (TPS)), which measures only the proportion of tumor cells that stain the membrane for PD-L1 (% of TC), e.g. As determined by FDA approved testing or CE marking testing. In some embodiments, TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 as determined by staining immune cells (%IC). In some embodiments, the IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1, as determined by staining tumor cells (%TC). In some embodiments, TC > 1%. In some embodiments, the chemotherapeutic agent administered is a standard-of-care chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is selected from 5-fluorouracil (5-FU), capecitabine (XELODA ^® ), irinotecan (CAMPTOSAR ^® ), trifluridine (Trifluridine), and tipiracil ( Combinations of chemotherapeutic agents of tipiracil) (LONSURF ^® ) and platinum drugs (eg cisplatin, carboplatin, oxaliplatin) or combinations thereof. In some embodiments, leucovorin is administered in combination with chemotherapy. In some embodiments, the combination of chemotherapeutic agents is a combination of folinic acid (folinic acid), 5-fluorouracil, and irinotecan (FOLFIRI). In some embodiments, the combination of chemotherapeutic agents is folinic acid (folinic acid), 5-fluorouracil, and oxaliplatin (FOLFOX). In some embodiments, the combination of chemotherapeutic agents is folinic acid (folinic acid), 5-fluorouracil, oxaliplatin, and irinotecan (FOLFOXIRI). In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic and unresectable colorectal cancer in a second-line advanced/metastatic setting, wherein the patient is administered Tracicill, chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor that is microsatellite instability-high (MSI-H) or mismatch repair-deficient (dMMR)-type as confirmed by FDA-approved testing or CE-mark testing. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a composite positive score (CPS) > 10. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1, as determined by staining tumor cells (%TC). In some embodiments, TC > 1%. In some embodiments, the chemotherapeutic agent administered is a standard-of-care chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is selected from 5-fluorouracil (5-FU), capecitabine (XELODA ^® ), irinotecan (CAMPTOSAR ^® ), trifluridine, and tipiracil (LONSURF ^® ) Combinations of chemotherapeutic agents with platinum drugs (eg cisplatin, carboplatin, oxaliplatin) or combinations thereof. In some embodiments, folate is administered in combination with chemotherapy. In some embodiments, the combination of chemotherapeutic agents is folinic acid (folinic acid), 5-fluorouracil, and irinotecan (FOLFIRI). In some embodiments, the combination of chemotherapeutic agents is folinic acid (folinic acid), 5-fluorouracil, and oxaliplatin (FOLFOX). In some embodiments, the combination of chemotherapeutic agents is folinic acid (folinic acid), 5-fluorouracil, oxaliplatin, and irinotecan (FOLFOXIRI). In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic and unresectable colorectal cancer in a second-line advanced/metastatic setting, wherein the patient is administered Tracicill, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor that is microsatellite instability-high (MSI-H) or mismatch repair-deficient (dMMR)-type as confirmed by FDA-approved testing or CE-mark testing. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a composite positive score (CPS) > 10, as determined by an FDA-approved test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1, as determined by staining tumor cells (%TC). In some embodiments, TC > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic and unresectable colorectal cancer in a second-line advanced/metastatic setting, wherein the patient is administered Tracicill, PD-1 or PD-L1 inhibitors, chemotherapy agents, and vascular endothelial growth factor (VEGF) inhibitors. In some embodiments, the VEGF inhibitor is selected from bevacizumab (Avastin), Ramucirumab (Cyramza) or ziv-aflibercept (Zaltrap) . In some embodiments, the patient has a tumor that is microsatellite instability-high (MSI-H) or mismatch repair-deficient (dMMR)-type as confirmed by an FDA-approved test. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test. In some embodiments, the patient has a tumor expressing PD-L1 and a composite positive score (CPS) > 10, as determined by an FDA-approved test or a CE-marked test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1, as determined by staining tumor cells (%TC). In some embodiments, TC > 1%. In some embodiments, the chemotherapeutic agent administered is a standard-of-care chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is selected from 5-fluorouracil (5-FU), capecitabine (XELODA ^® ), irinotecan (CAMPTOSAR ^® ), trifluridine, and tipiracil (LONSURF ^® ) Combinations of chemotherapeutic agents with platinum drugs such as cisplatin, carboplatin or oxaliplatin, or combinations thereof. In some embodiments, folate is administered in combination with chemotherapy. In some embodiments, the combination of chemotherapeutic agents is folinic acid (folinic acid), 5-fluorouracil, and irinotecan (FOLFIRI). In some embodiments, the combination of chemotherapeutic agents is folinic acid (folinic acid), 5-fluorouracil, and oxaliplatin (FOLFOX). In some embodiments, the combination of chemotherapeutic agents is folinic acid (folinic acid), 5-fluorouracil, oxaliplatin, and irinotecan (FOLFOXIRI). In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy. In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic and unresectable colorectal cancer in a second-line advanced/metastatic setting, wherein the patient is administered Tracicill, PD-1 or PD-L1 inhibitors, and vascular endothelial growth factor (VEGF) inhibitors. In some embodiments, the VEGF inhibitor is selected from bevacizumab (Avastin ^® ), ramucirumab (Cyramza ^® ), or ziv-aflibercept (Zaltrap ^® ). In some embodiments, the patient has a tumor that is microsatellite instability-high (MSI-H) or mismatch repair-deficient (dMMR)-type as confirmed by an FDA-approved test. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test. In some embodiments, the patient has a tumor expressing PD-L1 and a composite positive score (CPS) > 10, as determined by an FDA-approved test or a CE-marked test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1, as determined by staining tumor cells (%TC). In some embodiments, TC > 1%.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic and unresectable colorectal cancer in a second-line advanced/metastatic setting, wherein the patient is administered Tracicill, PD-1 or PD-L1 inhibitors, and epidermal growth factor (EGFR) inhibitors. In some embodiments, the EGFR inhibitor is selected from cetuximab (Erbitux ^® ) or panitumumab (Vectibix ^® ). In some embodiments, the patient has a tumor that is microsatellite instability-high (MSI-H) or mismatch repair-deficient (dMMR)-type as confirmed by an FDA-approved test. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a composite positive score (CPS) > 10, as determined by an FDA-approved test or a CE-marked test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1, as determined by staining tumor cells (%TC). In some embodiments, TC > 1%.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic or unresectable colorectal cancer in a first-line or second-line advanced/metastatic setting, wherein the patient is given Triracicill, a PD-1 or PD-L1 inhibitor, and an oncolytic virus are administered. In some embodiments, the oncolytic virus is selected from the group consisting of oncolytic adenovirus, oncolytic HSV-1, oncolytic Leo virus, oncolytic pox virus, oncolytic Newcastle disease virus, oncolytic measles virus, oncolytic Seneca Valley virus, Japanese hemagglutinating virus envelope (HVJ-E) virus, oncolytic gamma herpes virus, oncolytic parvovirus or oncolytic retrovirus. In some embodiments, the oncolytic virus is Palerip. In some embodiments, the patient has a tumor that is microsatellite instability-high (MSI-H) or mismatch repair-deficient (dMMR)-type as confirmed by an FDA-approved test. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test. In some embodiments, the patient has a tumor expressing PD-L1 and a composite positive score (CPS) > 10, as determined by an FDA-approved test. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1, as determined by staining tumor cells (%TC). In some embodiments, TC > 1%.

Urothelial Carcinoma In one aspect, the modified treatment approach described herein is administered to patients with locally advanced, recurrent unresectable or metastatic urothelial carcinoma of the bladder in the first-line advanced/metastatic setting ( mUC), and wherein the patient is administered triaciclib, one or more chemotherapeutic agents, PD-1 or PD-L1 inhibitors and selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or Other immune checkpoint inhibitors of CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 and a composite positive score (CPS) > 10. In some embodiments, the patient has a tumor expressing PD-L1 with an IC >5%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 50%. In some embodiments, the chemotherapeutic agent administered is a standard-of-care chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is selected from the group consisting of enfortumab vedotin, gemcitabine, cisplatin, carboplatin, eribulin, methotrexate, vinblastine, vinflunine Chemotherapy of (vinflunine), doxorubicin, epirubicin, ifosfamide, paclitaxel, nab-paclitaxel, docetaxel or any pharmaceutically acceptable salt thereof or combinations thereof agent. In some embodiments, the chemotherapeutic agent is gemcitabine and carboplatin or cisplatin. In some embodiments, the chemotherapeutic agent is methotrexate, vinblastine, doxorubicin, and cisplatin. In some embodiments, the chemotherapeutic agent is paclitaxel, gemcitabine and cisplatin. In some embodiments, the chemotherapeutic agents are methotrexate, carboplatin, and vinblastine. In some embodiments, the chemotherapeutic agent is gemcitabine and paclitaxel or docetaxel. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved treatment methods described herein are administered to patients with locally advanced, recurrent unresectable or metastatic bladder urothelial carcinoma (mUC) in a second-line advanced/metastatic setting A patient, and wherein the patient is administered triraciclib, one or more chemotherapeutic agents, a PD-1 or PD-L1 inhibitor, and a TIGIT inhibitor, a TIM-3 inhibitor, a LAG-3 inhibitor, or a CD73 inhibitor other immune checkpoint inhibitors. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 5%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 25%. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the chemotherapeutic agent administered is a standard-of-care chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is selected from the group consisting of paclitaxel, docetaxel, vinflunine, gosatuzumab, pemetrexed, paclitaxel, nab-paclitaxel, docetaxel, gemcitabine , ifosfamide and oxaliplatin or any pharmaceutically acceptable salt thereof or a chemotherapeutic agent of a combination thereof. In some embodiments, the chemotherapeutic agent is gemcitabine and carboplatin or cisplatin. In some embodiments, the chemotherapeutic agent is methotrexate, vinblastine, doxorubicin, and cisplatin. In some embodiments, the chemotherapeutic agent is paclitaxel, gemcitabine and cisplatin. In some embodiments, the chemotherapeutic agents are methotrexate, carboplatin, and vinblastine. In some embodiments, the chemotherapeutic agent is gemcitabine and paclitaxel or docetaxel. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved treatment methods described herein are administered to patients with locally advanced, recurrent unresectable or metastatic bladder urothelial carcinoma (mUC) in a second-line advanced/metastatic setting A patient, and wherein the patient is administered triraciclib, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors, or CD73 inhibitors . In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 5%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 25%. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In one aspect, the improved methods of treatment described herein are administered to patients with locally advanced, recurrent unresectable or metastatic urothelial bladder carcinoma (MUC) in a first-line or second-line advanced/metastatic setting. ), wherein the patient is administered triaciclib, a PD-1 or PD-L1 inhibitor, and an oncolytic virus. In some embodiments, the oncolytic virus is selected from the group consisting of oncolytic adenovirus, oncolytic HSV-1, oncolytic Leo virus, oncolytic pox virus, oncolytic Newcastle disease virus, oncolytic measles virus, oncolytic Seneca Valley virus, Japanese hemagglutinating virus envelope (HVJ-E) virus, oncolytic gamma herpes virus, oncolytic parvovirus or oncolytic retrovirus. In some embodiments, the oncolytic virus is Palerip. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test. In some embodiments, the patient has a tumor expressing PD-L1 and a composite positive score (CPS) > 10, as determined by an FDA-approved test. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 5%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 25%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 50%. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient is also administered a drug selected from the group consisting of paclitaxel, docetaxel, vinflunine, gosatuzumab, pemetrexed, paclitaxel, nab-paclitaxel, docetaxel, Gemcitabine, ifosfamide, enfortuzumab-vedotin, eribulin, methotrexate, vinblastine and oxaliplatin, doxorubicin, epirubicin, ifosfamide A chemotherapeutic agent of an amine or any pharmaceutically acceptable salt thereof or a combination thereof.

Solid Tumors In some aspects, the improved treatment methods described herein are administered to patients with locally advanced, recurrent unresectable or metastatic solid tumors in a first-line or second-line advanced/metastatic setting, and wherein the patient is administered triraciclib, one or more chemotherapeutic agents, a PD-1 or PD-L1 inhibitor, and another agent selected from a TIGIT inhibitor, a TIM-3 inhibitor, a LAG-3 inhibitor, or a CD73 inhibitor Immune checkpoint inhibitors. In some embodiments, the solid tumor is selected from cervical cancer, head and neck squamous cell carcinoma (SCCHN), cutaneous squamous cell carcinoma (cSCC), Merkel cell carcinoma, basal cell carcinoma, small cell lung cancer ( SCLC), melanoma, malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, high microsatellite instability or mismatch repair deficient cancer, endometrial cancer, high tumor mutation burden (TMB-H) cancer, Cancer of the stomach, gastroesophageal junction, adenocarcinoma of the esophagus, or cancer of the esophagus. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 5%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 25%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 50%. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 and a composite positive score (CPS) > 10. In some embodiments, the chemotherapeutic agent administered is a standard-of-care chemotherapeutic agent. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In some aspects, the improved treatment methods described herein are administered to patients with locally advanced, recurrent unresectable or metastatic solid tumors in a first-line or second-line advanced/metastatic setting, and wherein Triracicill, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors, or CD73 inhibitors are administered to the patient. In some embodiments, the solid tumor is selected from cervical cancer, head and neck squamous cell carcinoma (SCCHN), cutaneous squamous cell carcinoma (cSCC), Merkel cell carcinoma, basal cell carcinoma, small cell lung cancer (SCLC), melanoma Malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, high microsatellite instability or mismatch repair deficient cancer, endometrial cancer, high tumor mutation burden (TMB-H) cancer, gastric cancer, gastroesophageal cancer Carcinoma of the junction, adenocarcinoma of the esophagus, or cancer of the esophagus. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 1%. In some embodiments, the patient has a tumor expressing PD-L1 with an IC > 5%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 25%. In some embodiments, the patient has a tumor expressing PD-L1 with a TC > 50%. In some embodiments, the patient has a tumor expressing PD-L1 and a TPS > 1%. In some embodiments, the patient has a tumor expressing PD-L1 and a composite positive score (CPS) > 10. In some embodiments, the patient has previously received a PD-1 or PD-L1 inhibitor and experienced disease progression. In some embodiments, the patient is not receiving PD-1 or PD-L1 therapy.

In some aspects, the improved treatment methods described herein are administered to patients with locally advanced, recurrent unresectable or metastatic solid tumors in a first-line or second-line advanced/metastatic setting, wherein the Patients were administered triraciclib, a PD-1 or PD-L1 inhibitor, and an oncolytic virus. In some embodiments, the solid tumor is selected from cervical cancer, head and neck squamous cell carcinoma (SCCHN), cutaneous squamous cell carcinoma (cSCC), Merkel cell carcinoma, basal cell carcinoma, small cell lung cancer (SCLC), melanoma Malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, high microsatellite instability or mismatch repair deficient cancer, endometrial cancer, high tumor mutation burden (TMB-H) cancer, gastric cancer, gastroesophageal cancer Carcinoma of the junction, adenocarcinoma of the esophagus, or cancer of the esophagus. In some embodiments, the oncolytic virus is selected from the group consisting of oncolytic adenovirus, oncolytic HSV-1, oncolytic Leo virus, oncolytic pox virus, oncolytic Newcastle disease virus, oncolytic measles virus, oncolytic Seneca Valley virus, Japanese hemagglutinating virus envelope (HVJ-E) virus, oncolytic gamma herpes virus, oncolytic parvovirus or oncolytic retrovirus. In some embodiments, the oncolytic virus is Palerip. In some embodiments, the patient has a tumor expressing PD-L1. In some embodiments, chemotherapeutic agents are also administered to the patient.

Improved Patient Outcomes Administration of the treatment regimens described herein to the subgroups of patients described herein can provide enhanced anti-tumor efficacy in the patients. In some embodiments, administration of the treatment regimens described herein provides improved progression-free survival (PFS) and/or overall survival (OS) in the specific patient subgroups described above. In some embodiments, the PFS improvement is based on Response Evaluation Criteria in Solid Tumors 1.1 (RECIST 1.1).

In some embodiments, administration of the treatment regimens described herein to the aforementioned patient subgroups improves myeloprotection of hematopoietic stem/progenitor cells (HSPCs) and immune effector cells (eg, lymphocytes, including T lymphocytes). In some embodiments, administration of the treatment regimens described herein to the aforementioned patient subgroups reduces chemotherapy-induced myelosuppression (CIM). In some embodiments, administration of the treatment regimens described herein provides myeloprotection of the neutrophil lineage in patients compared to those receiving chemotherapy without treracilib. In some embodiments, administration of the treatment regimens described herein reduces the duration of severe (grade 4) neutropenia.

In some embodiments, administration of the treatment regimens described herein reduces severe neutropenia events, reduces granule colony stimulating factor (G-CSF) therapy, or reduces febrile neutropenia (FN ) adverse event (AE). In some embodiments, administration of a therapeutic regimen described herein reduces a grade 3 or 4 hemoglobin drop laboratory value, red blood cell (RBC) infusion, or erythropoiesis-stimulating agent (ESA) administration. In some embodiments, administration of the therapeutic regimens described herein reduces laboratory values for grade 3 or 4 platelet count depression and/or the number of platelet transfusions. In some embodiments, administration of the treatment regimens described herein reduces Grade 3 or 4 hematology laboratory values.

In some embodiments, administration of the treatment regimens described herein reduces all-cause dose reduction or cycle delay and the relative dose intensity for chemotherapy. In some embodiments, administration of the therapeutic regimens described herein reduces i) hospitalizations, including but not limited to, from all etiologies, febrile neutropenia/neutropenia, anemia/RBC infusion , thrombocytopenia/bleeding and infection); or ii) antibiotic use, including but not limited to intravenous (IV), oral, and oral and intravenous administration of antibiotics.

In some embodiments, administration of the treatment regimens described herein reduces chemotherapy-induced fatigue (CIF) in the patient. In some embodiments, lowering CIF reduces time to first confirmed fatigue exacerbation (TTCD-Fatigue), as measured by Functional Assessment of Cancer Therapy-Fatigue (FACIT-F).

In some embodiments, administration of the treatment regimens described herein improves one or more of: Functional Assessment of Cancer Therapy-General (FACT-G) Domain Score (Physical, Social/Family, Emotional, and Functional Well-being Functional Assessment of Cancer Therapy-Anemia (FACT-An); 5-Scale EQ-5D (EQ-5D-5L); Patient Global Impression of Change (PGIC) Fatigue Item; or Patient Global Impression of Severity (PGIS) Fatigue Item.

CROSS REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application 63/238,739, filed August 30, 2021, and U.S. Provisional Application 63/399,977, filed August 22, 2022; for all purposes , the entire contents of each of these applications are incorporated herein by reference.

Definitions Unless stated otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. In this specification, the singular also includes the plural unless the context clearly dictates otherwise. Although methods and materials similar or equivalent to those disclosed herein can be used in the practice or testing of the present application, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are hereby incorporated by reference. References cited herein are not admitted to be prior art to the claimed application. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

Compounds are described using standard nomenclature. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

In some embodiments of each compound described herein, unless the context clearly excludes, the compound may be a racemate, an enantiomer, a mixture of enantiomers, a diastereomer, a diastereomer Enantiomeric mixtures, tautomers, N-oxides or forms of isomers (eg rotamers) are as if each explicitly stated.

The terms "a and an" do not denote a limitation of quantity, but mean that there is at least one of the referenced items. The term "or" means "and/or". Recitation of ranges of values are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All range endpoints are included within that range and are independently combinable. All methods described herein can be performed in a suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The examples, or exemplary language (such as "such as"), are intended merely to better illuminate the invention and do not pose a limitation on the scope of the invention unless stated otherwise.

As used herein, an "effective amount" means an amount that provides a therapeutic or prophylactic benefit.

As used herein, the term "treating" a disease means reducing the frequency or severity of at least one sign or symptom of a disease, disorder, or side effect experienced by a patient (i.e., palliative treatment) or reducing the amount of time experienced by a patient by administering a therapeutic agent. The cause or effect of a disease, disorder or side effect (ie disease modifying treatment).

Throughout this disclosure, various aspects of this invention can be presented in a range format. It should be understood that the presentation in range format is for convenience only and should not be construed as limitations on the scope of the invention. The statement of a range should be considered as specifically disclosing all possible subranges as well as individual values within that range. For example, a stated range of, for example, 1 to 6 should be considered to have specifically disclosed sub-ranges (eg, 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc.) and Individual values within the range (eg, 1, 2, 2.7, 3, 4, 5, 5.3, and 6). This applicability is independent of breadth of scope.

As used herein, a "pharmaceutical composition" is a composition that includes at least one active agent and at least one other substance, such as a carrier. A "pharmaceutical combination" is a combination of at least two active agents, which may be combined in a single dosage form or provided together in separate dosage forms, together with instructions, which are intended to be used together for the treatment of any of the conditions described herein.

As used herein, "pharmaceutically acceptable salts" are derivatives of the disclosed compounds wherein the parent compound is modified by making inorganic or organic, non-toxic, acid or base addition salts thereof. Salts of compounds of the present invention can be synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods. Typically, such salts can be prepared by reacting the free acid form of such compounds with a stoichiometric amount of an appropriate base such as Na, Ca, Mg or K hydroxides, carbonates, bicarbonates or the like, or Prepared by reacting the free base form of these compounds with a stoichiometric amount of the appropriate acid. These reactions are usually carried out in water or organic solvents or a mixture of both. In general, non-aqueous media are typical, such as diethyl ether, ethyl acetate, ethanol, isopropanol, or acetonitrile, where available. The salts of the compounds of the present invention further include solvates of the compounds and salts of the compounds.

Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines, basic or organic salts of acidic residues such as carboxylic acids, and the like. Pharmaceutically acceptable salts include conventional non-toxic and quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. By way of example, customary non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, and the like; and salts from organic acids such as acetic, Propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzene Formic acid, salicylic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, sulfanilic acid, 2-acetyloxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, Isethionic acid, HOOC-(CH2)n-COOH (where n is 0-4), and the like; or using a different acid former that yields the same counterion. Lists of other suitable salts can be found, for example, in Remington's Pharmaceutical Sciences, 17th Ed., Mack Publishing Company, Easton, Pa., p. 1418 (1985). Where the methods described herein demonstrate the administration of a particular compound, it is understood that, as an embodiment, administration of pharmaceutically acceptable salts of the compounds is contemplated, where applicable.

As used herein, the term "prodrug" means a compound that is converted to the parent drug when administered to a host in vivo. As used herein, the term "parent drug" means a drug described herein that can be used to treat any of the conditions described herein or to control or improve any physiology or pathology in a host (usually a human) that is described herein. Any chemical compound associated with an underlying cause or symptom of a disorder. Prodrugs can be used to achieve any desired effect, including enhancing the properties of the parent drug or improving the pharmaceutical or pharmacokinetic properties of the parent drug. Prodrug strategies exist that are selected to modulate the in vivo production conditions of the parent drug and are considered encompassed herein. Non-limiting examples of prodrug strategies include covalent attachment of removable groups or moieties of removable groups, such as, but not limited to, acylation, phosphorylation, phosphonation, phosphoramidate derivatives, inter alia, Amidation, reduction, oxidation, esterification, alkylation, other carboxyl derivatives, sulfinyl or sulfenyl derivatives, carbonylation or anhydrides.

The term "carrier" as applied to the pharmaceutical composition/composition of the present invention refers to a diluent, excipient or vehicle provided with the active compound.

"Pharmaceutically acceptable excipient" means an excipient that can be used in the preparation of a pharmaceutical composition/combination that is generally safe and not biologically and otherwise unsuitable for administration to a host, typically a human.

In a non-limiting example, treracicill may be used in a form in which at least one desired isotopic atom is substituted in an amount greater than the natural isotopic abundance (ie, enriched). Isotopes are atoms having the same atomic number but different mass numbers (ie, the same number of protons but different numbers of neutrons).

Examples of isotopes that may be incorporated into triraciclib used in the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine and iodine, such as 2H, 3H, 11C, 13C, 14C, 15N, 18F 31P, 32P, 35S, 36CI and 125I. In one non-limiting example, isotopically labeled compounds can be used in metabolic studies (14C), reaction kinetics studies (such as 2H or 3H), detection or imaging techniques (such as positron emission tomography (PET) or single Photon emission computerized tomography (SPECT), including drug or matrix tissue distribution analysis) or radiation therapy of patients. In particular, 18F-labeled compounds may be particularly desirable for PET or SPECT studies. Isotopically-labeled compounds of the invention and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the reaction schemes or in the Examples and Preparations below and by substituting a readily available isotopically-labeled reagent for a non-isotopically-labeled reagent.

By way of general example and not limitation, hydrogen isotopes such as deuterium ( ^2H ) and tritium ( ^3H ) can be used anywhere in the illustrated structures to achieve the desired result. Alternatively or additionally, carbon isotopes such ^{as13C and14C} may be used. Isotopic substitution (eg, deuterium substitution) may be partial or complete. Partial deuterium substitution means that at least one hydrogen is replaced by deuterium. In certain embodiments, the isotope is 90%, 95%, or 99% or more enriched isotope. In a non-limiting example, deuterium is 90, 95 or 99% enriched at the desired location.

In a non-limiting example, a CDK4/6 inhibitor, chemotherapy, or checkpoint inhibitor may be used in a form in which at least one desired isotopic atom is substituted in an amount greater than the natural isotopic abundance (ie, enriched). Isotopes are atoms having the same atomic number but different mass numbers (ie, the same number of protons but different numbers of neutrons).

Triraciclib or another CDK4/6 inhibitor described herein for use in the present invention may form solvates with solvents, including water. Accordingly, in one non-limiting embodiment, the invention encompasses the use of the solvated forms of the compounds. The term "solvate" refers to a molecular complex of a compound of the invention (including salts thereof) with one or more solvent molecules. Non-limiting examples of solvents are water, ethanol, dimethylsulfoxide, acetone, and other common organic solvents. The term "hydrate" refers to a molecular complex comprising a compound of the present invention and water. Pharmaceutically acceptable solvates of the present invention include those wherein the solvent may be isotopically substituted, eg D ₂ O, d ₆ -acetone, d ₆ -DMSO. Solvates may be in liquid or solid form.

As generally considered herein, the term "hematopoietic stem/progenitor cells" (HSPC) includes, but is not limited to, long-term hematopoietic stem cells (LT-HSC), short-term hematopoietic stem cells (ST-HSC), hematopoietic progenitor cells (HPC), multipotent Progenitors (MPPs), Oligodendritic Preprogenitors (OPPs), Monocytes, Granulocytes, Common Myeloid Progenitors (CMP), Common Lymphoid Progenitors (CLP), Granules-Monocytes Glomerular progenitors (GMP), granulocytic progenitors, monocytic progenitors and megakaryocyte-erythroid progenitors (MEP), megakaryocyte progenitors, erythroid progenitors, HSC/MPP (CD45dim/CD34+/CD38-) , OPP (CD45dim/CD34+/CD38+), monocytic progenitors (CD45+/CD14+/CD11b+), granulocytic progenitors (CD45+/CD14-/CD11b+), erythroid progenitors (CD45-/CD71+) and megakaryocyte progenitors Cells (CD45+/CD61+).

The term "immune effector cells" generally refers to immune cells that perform one or more specific functions. Immune effector cells are known in the art and include, for example but not limited to, T cells (including naive T cells, memory T cells, activated T cells (T helper (CD4+) and cytotoxic T cells (CD8+)), TH1 Activated T cells, TH2 activated T cells, TH17 activated T cells), naive B cells, memory B cells, plasmablasts, dendritic cells, monocytes and natural killer (NK) cells.

The "patient," "host" or "subject" treated is typically a human patient, although it is understood that the methods described herein may be usefully applied to other animals (eg, mammals). More specifically, the term patient may include animals used in assays, such as those used in preclinical testing, including but not limited to mice, rats, monkeys, dogs, pigs, and rabbits; and domestic pigs (wild boars and hogs), ruminants, horses, poultry, cats, cattle, rodents, dogs and the like.

In some embodiments, the term "CDK4/6 replication independent cancer" refers to a cancer for which the activity of CDK4/6 is not significantly required for replication. This type of cancer is often, but not always, characterized (eg, cells exhibit) increased levels of CDK2 activity or expression of retinoblastoma tumor suppressor proteins or retinoblastoma family member proteins such as, but not limited to, p107 and p130 Reduced performance. The degree of increased CDK2 activity or decreased or absent expression of retinoblastoma tumor suppressor proteins or retinoblastoma family member proteins may be higher or lower than, for example, normal cells. In some embodiments, the increased level of CDK2 activity can be associated with (eg, can result from or be observed concurrently with) amplification or overexpression of the MYC proto-oncogene. In some embodiments, the increased level of CDK2 activity can be associated with overexpression of cyclin El, cyclin E2, or cyclin A.

In some embodiments, the term "CDK4/6 replication-dependent cancer" refers to a cancer that requires the activity of CDK4/6 for replication or proliferation or whose growth can be inhibited by the activity of a selective CDK4/6 inhibitor. Cancers and disorders of this type can be characterized (eg, cells exhibited by) the presence of a functional retinoblastoma (Rb) protein. These cancers and disorders are classified as Rb positive. Rb-positive abnormal cell proliferation disorders and variations of this term as used herein refer to disorders or diseases caused by uncontrolled or abnormal cell division, characterized by the presence of functional retinoblastoma protein and may include cancer.

As used herein, the term "immune checkpoint inhibitor (ICI)" refers to therapies that target immune checkpoint proteins that are key to the immune system when expressed to attenuate the immune response to immune stimulants regulator. Some cancers express ligands for checkpoint inhibitors and can protect themselves from attack by binding to immune checkpoint targets. ICIs block inhibitory checkpoints, thereby restoring immune system function. ICIs include those targeting immune checkpoint proteins such as: PD-1, PD-1 ligand-1 (PD-L1), PD-1 ligand-2 (PD-L2), CTLA-4, LAG-3 , TIM-3, cluster of differentiation 73 (CD73) and V domain Ig inhibitor of T cell activation (VISTA), B7-H3/CD276, indoleamine 2, 3-dioxygenase (IDO), killer immunoglobulin protein-like receptor (KIR), carcinoembryonic antigen cell adhesion molecule (CEACAM) (such as CEACAM-1, CEACAM-3, and CEACAM-5), sialic acid-binding immunoglobulin-like lectin 15 (Siglec-15), with Ig and ITIM domain T cell immune receptor (TIGIT) and B and T lymphocyte attenuation protein (BTLA). Immune checkpoint inhibitors are known in the art.

PD-L1 Status As provided herein, the intended treatment for non-small cell lung cancer (NSCLC), triple-negative breast cancer (TNBC), colorectal cancer (CRC), metastatic urothelial carcinoma (mUC), or another solid tumor is typically PD-L1 positive. In alternative embodiments, the NSCLC, TNBC, colorectal, mUC or another solid tumor to be treated is PD-L1 negative.

PD-L1 is a transmembrane protein that down-regulates the immune response by binding to its two inhibitory receptors: programmed death-1 (PD-1) and B7.1. PD-1 is an inhibitory receptor expressed on T cells following T cell activation, which persists in chronically stimulated states (such as chronic infection or cancer) (Blank, C and Mackensen, A, Contribution of the PD-L1 /PD-1 pathway to T-cell exhaustion: an update on implications for chronic infections and tumor evasion. Cancer Immunol Immunother, 2007. 56(5): p. 739-745). The combination of PD-L1 and PD-1 inhibits T cell proliferation, interleukin production, and cytolytic activity, resulting in functional inactivation or T cell exhaustion. B7.1 is a molecule expressed on antigen presenting cells and activated T cells. Binding of PD-L1 to B7.1 on T cells and antigen-presenting cells can mediate downregulation of immune responses, including inhibition of T cell activation and cytokine production (see Butte MJ, Keir ME, Phamduy TB et al., Programmed death- 1 ligand 1 interacts specifically with the B7-1 costimulatory molecule to inhibit T cell responses. Immunity. 2007; 27(1): 111-122). PD-L1 expression has been observed in immune cells and tumor cells. See Dong H, Zhu G, Tamada K, Chen L. B7-H1, a third member of the B7 family, co-stimulates T-cell proliferation and interleukin-10 secretion. Nat Med. 1999; 5(12): 1365- 1369; Herbst RS, Soria JC, Kowanetz M, et al. Predictive correlates of response to the anti-PD-L1 antibody MPDL3280A in cancer patients. Nature. 2014; 515(7528): 563-567. Aberrant expression of PD-L1 on tumor cells has been reported to impede anti-tumor immunity, thereby causing immune evasion.

PD-L1 expression can be determined by methods known in the art. For example, PD-L1 IHC 22C3 pharmDx (FDA-approved in vitro diagnostic immunohistochemistry (IHC) developed by Dako and Bristol-Meyers Squibb as a companion test for pembrolizumab ( ^KEYTRUDA® ) therapy can be used ) test) to detect PD-L1 expression. This system uses a single mouse anti-PD-L1 clone 22C3 PD-L1 and EnVision FLEX visualization system on the Autostainer Link 48 platform to detect PD- Qualitative analysis of L1. Expressive content can be measured using the Composite Positive Score (CPS). This scoring method evaluates the ratio of the number of PD⁠-⁠L1 staining cells (tumor cells, lymphocytes, macrophages) relative to all viable tumor cells. Use CPS to evaluate PD⁠-⁠L1 manifestations in metastatic or unresectable, recurrent HNSCC, advanced esophageal or GEJ cancer, metastatic urothelial carcinoma (mUC), colorectal cancer, advanced Cervical cancer and advanced triple-negative breast cancer. Expressed content can be measured using the Tumor Proportion Score (TPS), which measures the percentage of viable tumor cells exhibiting partial or complete membrane staining. Staining can demonstrate PD-L1 expression from 1% to 100%. Use TPS to evaluate PD⁠-⁠L1 expression in advanced NSCLC, metastatic urothelial carcinoma (mUC), colorectal cancer, and other solid tumors.

PD-L1 IHC 28-8 pharmDx, an FDA-approved in vitro diagnostic immunohistochemistry (IHC) test developed by Dako and Merck as a companion test for nivolumab (OPDIVO ^® ) therapy, can also be used to Detection of PD-L1 expression. This qualitative assay detects PD-L1 in formalin-fixed paraffin-embedded (FFPE) human cancer tissue using a single rabbit anti-PD-L1 clone 28-8 and the EnVision FLEX visualization system on the Autostainer Link 48 platform. PD-L1 protein expression results are reported as the percentage of viable tumor cells exhibiting any degree of membrane staining (%TC). Use %TC to evaluate PD-L1 expression in advanced non-squamous NSCLC, colorectal cancer (CRC), metastatic urothelial carcinoma (mUC), and other solid tumors (e.g., cervical cancer, head and neck squamous cell carcinoma). squamous cell carcinoma (SCCHN), squamous cell carcinoma of the skin (cSCC), Merkel cell carcinoma, basal cell carcinoma, small cell lung cancer (SCLC), melanoma, malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, high Microsatellite instability or mismatch repair deficient cancer, endometrial cancer, tumor mutation high burden (TMB-H) cancer, gastric cancer, gastroesophageal junction cancer, esophageal adenocarcinoma, and esophageal cancer).

Other commercially available assays for PD-L1 detection include the Ventana SP263 assay using a monoclonal rabbit anti-PD-L1 clone SP263 (developed by Ventana in conjunction with AstraZeneca) and the Ventana SP142 assay using a rabbit monoclonal anti-PD-L1 clone SP142 (developed by Ventana is jointly developed by Genentech/Roche). The VENTANA PD-L1 (SP263) assay was used on the VENTANA BenchMark ULTRA instrument using rabbit monoclonal anti-PD-L1 clone SP263 for the evaluation of formalin-fixed paraffin-embedded (FFPE) stained with the OptiView DAB IHC kit Qualitative immunohistochemical analysis of PD-L1 protein in urothelial carcinoma tissues. PD-L1 status was determined by the percentage of tumor cells above background with any membrane staining (%TC) or by the percentage of tumor-associated immune cells above background with staining at any intensity (IC%). IC% was determined using the percentage of tumor area occupied by any tumor-associated immune cells (immune cells present, ICPs), IC% being the area percentage of ICPs exhibiting PD-L1 positive immune cell staining. PD-L1 status can be considered high when any of the following conditions are met: ≥ 25% of tumor cells exhibit membrane staining; or ICP > 1% and IC% ≥ 25%; or ICP = 1% and IC+ = 100% . VENTANA PD-L1 (SP142) Assay is a qualitative immunohistochemical analysis performed on a BenchMark ULTRA instrument using rabbit monoclonal anti-PD-L1 clone SP142 for the evaluation of staining by the OptiView DAB IHC Detection Kit and the OptiView Amplification Kit Tumor cells and tumor-infiltrating immune cells in formalin-fixed paraffin-embedded (FFPE) tissues including metastatic urothelial carcinoma (mUC), triple-negative breast cancer (TNBC) and non-small cell lung cancer (NSCLC) Programmed death-ligand 1 (PD-L1) protein. Results can be used to determine viability of treatment with TECENTRIQ ^® (Roche/Genentech). Determination of PD-L1 status is indication-specific and assessment is based on the fraction of tumor area occupied by tumor-infiltrating immune cells expressing PD-L1 of any intensity (%IC) or the percentage of tumor cells expressing PD-L1 of any intensity (%TC). The cutoff value was ≥5% IC for mUC, ≥1% IC for TNBC and ≥50% TC or 10% IC for NSCLC.

In some embodiments, the NSCLC patient treated with one of the first-line or second-line treatment regimens described herein has confirmed PD-L1 status positive NSCLC. In some embodiments, patients treated with first-line or second-line regimens have PD-L1-stained tumor-infiltrating immune cells (IC) ≥ 10% or tumor cells (TC) ≥ 50% with a confirmed positive PD-L1 status NSCLC, as confirmed by in vitro diagnostic (IVD) assays such as Ventana SP-142 assays or other suitable assays. In some embodiments, the patient treated with the first-line or second-line regimen has NSCLC with proven PD-L1 status of ≥25% of tumor cells staining for PD-L1, as determined by in vitro diagnostic (IVD) analysis (e.g., Ventana SP-142 assay or other appropriate assay). In an alternative embodiment, the patient treated with the first-line or second-line regimen has NSCLC with a proven PD-L1 status of ≥1% tumor cell (TC) PD-L1 staining, such as by an FDA-approved test or CE mark determined by the test. In an alternative embodiment, the patient treated with the first-line or second-line regimen has NSCLC with a confirmed negative PD-L1 status.

In some embodiments, the TNBC patient treated with one of the first-line or second-line treatment regimens described herein has confirmed PD-L1 status positive TNBC. In some embodiments, the patient treated with the first-line or second-line regimen has a ratio of PD-L1 staining cells (tumor cells, lymphocytes, macrophages) to all viable tumor cells (CPS) ≥ 10% or PD -L1-stained tumor-infiltrating immune cells (IC) ≥1% of TNBC with confirmed PD-L1 status positive, as analyzed by in vitro diagnostic (IVD) analysis (e.g. Dako PD-L1-22C3 pharmDx kit or Ventana SP-142 analysis or another appropriate analysis). In some embodiments, patients treated with first-line or second-line treatment regimens have TNBC with confirmed PD-L1 status of ≥10% of tumor cells staining for PD-L1, as determined by in vitro diagnostic (IVD) analysis (e.g., Ventana SP-142 assay or other appropriate assay). In an alternative embodiment, the patient treated with the first-line or second-line regimen has TNBC with ≥ 1% immune cell PD-L1 staining with proven PD-L1 status, as determined by an FDA-approved test or a CE-marked test . In an alternate embodiment, the patient treated with the first-line or second-line regimen has TNBC with a confirmed negative PD-L1 status.

In some embodiments, a colorectal cancer (CRC) patient treated with one of the first-line or second-line treatment regimens described herein has microsatellite instability-high (MSI-H) demonstrated by FDA-approved testing or CE-mark testing or mismatch repair-deficient (dMMR)-proven tumors. In some embodiments, the CRC patient treated with one of the first-line or second-line treatment regimens described herein has confirmed PD-L1 status positive CRC. In some embodiments, patients treated with first-line or second-line regimens have PD-L1-stained tumor-infiltrating immune cells (IC) ≥ 1% or tumor cells (TC) ≥ 1% with confirmed positive PD-L1 status CRC, as confirmed by in vitro diagnostic (IVD) assays such as Ventana SP-142 assays or other suitable assays. In some embodiments, patients treated with first-line or second-line treatment regimens have a ratio of PD-L1 staining cells (tumor cells, lymphocytes, macrophages) to all viable tumor cells (CPS) ≥ 10%. Confirmed PD-L1 status CRC, as confirmed by in vitro diagnostic (IVD) assays (eg, Ventana SP-142 assay or other appropriate assay). In an alternative embodiment, patients treated with first-line or second-line regimens have CRC with proven PD-L1 status ≥ 1% of viable tumor cells (TPS) exhibiting partial or complete membrane staining of any intensity, as determined by As determined by FDA-approved tests or CE-marked tests. In an alternative embodiment, patients treated with first-line or second-line regimens have CRC with proven PD-L1 status of ≥1% tumor cell (TC) PD-L1 staining, as by FDA-approved testing or CE marking determined by the test. In an alternate embodiment, the patient treated with the first-line or second-line regimen has CRC with a confirmed negative PD-L1 status.

In some embodiments, mUC patients treated with first-line or second-line treatment regimens have confirmed PD-L1 status positive mUC. In some embodiments, patients treated with first-line or second-line regimens have PD-L1-stained tumor-infiltrating immune cells (IC) > 5% or tumor cells (TC) ≥ 50% with a confirmed positive PD-L1 status mUC, as demonstrated by in vitro diagnostic (IVD) assays, such as Ventana SP-142 assays or other suitable assays. In some embodiments, patients treated with first-line or second-line treatment regimens have mUC with confirmed PD-L1 status of > 25% tumor cell (TC) PD-L1 staining, as analyzed by in vitro diagnostic (IVD) (eg Ventana SP-263 assay or other suitable assay). In some embodiments, patients treated with first-line or second-line treatment regimens have mUC with confirmed PD-L1 status of ≥5% immune cell (IC) PD-L1 staining, as analyzed by in vitro diagnostic (IVD) (eg Ventana SP-142 assay or other suitable assay). In an alternative embodiment, patients treated with first-line or second-line regimens have mUC with a proven PD-L1 status of ≥1% tumor cell PD-L1 staining (%TC), as measured by an FDA-approved test or CE As determined by the flag test. In an alternative embodiment, patients treated with first-line or second-line regimens have mUC with proven PD-L1 status of ≥ 1% immune cell PD-L1 staining (%IC), as measured by an FDA-approved test or CE As determined by the flag test. In some embodiments, patients treated with first-line or second-line treatment regimens have a ratio of PD-L1 staining cells (tumor cells, lymphocytes, macrophages) to all viable tumor cells (CPS) ≥ 10%. Confirmed PD-L1 status positive mUC, as confirmed by in vitro diagnostic (IVD) assays (eg, Ventana SP-263 assay or another suitable assay). In an alternative embodiment, patients treated with first-line or second-line treatment regimens have mUC with demonstrated PD-L1 status ≥ 1% of viable tumor cells (TPS) exhibiting partial or complete membrane staining of any intensity, as determined by As determined by FDA-approved tests or CE-marked tests. In an alternate embodiment, the patient treated with the first-line or second-line regimen has mUC with confirmed negative PD-L1 status.

In some embodiments, the solid tumor patient treated with the first-line or second-line treatment regimen has a confirmed PD-L1 status positive solid tumor selected from the group consisting of: Cervical Cancer, Squamous Cell Carcinoma of the Head and Neck (SCCHN) , cutaneous squamous cell carcinoma (cSCC), Merkel cell carcinoma, basal cell carcinoma, small cell lung cancer (SCLC), melanoma, malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, high microsatellite instability or Mismatch repair deficient cancer, endometrial cancer, high tumor mutation burden (TMB-H) cancer, gastric cancer, gastroesophageal junction cancer, esophageal adenocarcinoma, or esophageal cancer. In some embodiments, the patient treated with the first-line or second-line treatment regimen has a ratio of PD-L1 staining cells (tumor cells, lymphocytes, macrophages) to all viable tumor cells (CPS) ≥ 1% by experience Proven PD-L1 status positive squamous cell carcinoma of the head and neck (SCCHN). In some embodiments, the patient treated with the first-line or second-line treatment regimen has a ratio of PD-L1 staining cells (tumor cells, lymphocytes, macrophages) to all viable tumor cells (CPS) ≥ 1% by experience Proven PD-L1 status-positive cervical cancer, as determined by FDA-approved testing or CE-mark testing. In some embodiments, patients treated with first-line or second-line treatment regimens have a ratio of PD-L1 staining cells (tumor cells, lymphocytes, macrophages) to all viable tumor cells (CPS) ≥ 10% Esophageal cancer with confirmed PD-L1 status positive, as determined by FDA-approved testing or CE-mark testing. In some embodiments, the patient treated with the first-line or second-line treatment regimen has gastroesophageal junction cancer with confirmed PD-L1 status of ≧1% of viable tumor cells (TPS) exhibiting partial or complete membrane staining of any intensity, As determined by FDA approval testing or CE marking testing.

CDK4/6 Status As provided herein, the NSCLC, TNBC, CRC, mUC or another solid tumor to be treated is negative for CDK4/6. In alternative embodiments, the NSCLC, TNBC, CRC, mUC or another solid tumor to be treated is positive for CDK4/6. In still other alternative embodiments, NSCLC, TNBC, CRC, mUC or another solid tumor is CDK4/6 indeterminate.

CDK4/6 replication-independent cancers often have abnormalities in the retinoblastoma gene (Rb1). The gene product of Rbl, the Rb protein, is a downstream target of CDK4/6. RB1 is often dysregulated in cancer cells by deletion, mutation, or epigenetic modification (resulting in loss of RB expression) and by aberrant CDK kinase activity (causing hyperphosphorylation and inactivation of RB function) (Chen et al., Novel RB1-Loss Transcriptomic Signature Is Associated with Poor Clinical Outcomes across Cancer Types. Clin Cancer Res. 2019; 25(14); Sherr, C.J. and McCormick, F. The RB and p53 pathways in cancer. Cancer Cell, 2002; 2:103 12.) . CCNE1/2 (cyclin E) is part of a parallel CDK4/6 pathway that provides functional redundancy and helps transition cells from G1 to S phase. Overexpression will reduce dependence on the CDK4/6 pathway, resulting in CDK4/6 independence (Turner et al., Cyclin E1 Expression and Palbociclib Efficacy in Previously Treated Hormone Receptor-Positive Metastatic Breast Cancer. J Clin Oncol. 2019; 37( 14): 1169-78.). Thus, tumors with CCNE1/2 amplification or RB loss can often be considered "CDK4/6 independent".

CDK4/6 Replication-Dependent The activity of CDK4/6 is required for the replication or proliferation of cancers. CDK 4/6 replication-dependent TNBC typically have an intact and functional Rb pathway and/increased CDK4/6 activator (cyclin D) expression and/or d-type cyclin activation signature (DCAF) (including CCND1 translocation, CCND1- 3 3'UTR loss and CCND2 or CCND3 amplification) (see Gong et al., Genomic aberrations that activate D-type cyclins are associated with enhanced sensitivity to the CDK4 and CDK5 inhibitor abemaciclib. Cancer Cell. 2017; 32(6): 761 -76). Tumors that are wild-type for RB and CCNE1/2 and have one of the aforementioned DCAFs are generally classified as "CDK4/6-dependent".

Tumors that cannot be classified as CDK4/6 replication-dependent or CDK4/6 replication-independent are often classified as "CDK4/6 indeterminate" because they cannot be demonstrated to be CDK4/6 dependent or independent.

In some embodiments, NSCLC, TNBC, CRC, mUC or other solid tumors are classified as CDK4/6 replication dependent. In some embodiments, NSCLC, TNBC, CRC, mUC or other solid tumors are classified as CDK4/6 replication independent. In some embodiments, NSCLC, TNBC, CRC, mUC, or other solid tumors are classified as CDK4/6 indeterminate.

Methods for determining CDK4/6 gene signature analysis are known in the art and involve the use of tumor tissue collected from a patient's biopsy (NSCLC, TNBC, CRC, mUC, or another solid tumor's primary or metastatic site), and Described in Shapiro GI. Genomic biomarkers predicting response to selective CDK4/6 inhibition: Progress in an elusive search. Cancer Cell. 2017; 32(6): 721-3 and Gong et al., Genomic aberrations that activate D-type cyclins are associated with enhanced sensitivity to the CDK4 and CDK5 inhibitor abemaciclib. Cancer Cell. 2017; 32(6): 761-76.

In some embodiments, triacillib (on a regimen with specific timing) is received in combination with an effective amount of a PD-1 or PD-L1 checkpoint inhibitor and an effective amount of another immune checkpoint inhibitor (ICI) (selected from a group with Ig and ITIM domain T cell immune receptor (TIGIT) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint inhibitors or cluster of differentiation (CD) 73 inhibitors) have CDK4/6-independent NSCLC with at least one of the following characteristics: a. CCNE1 amplification; b. CCNE2 amplification; or c. Loss of retinoblastoma protein 1 (Rb1), which is defined as i) a homozygous deletion, ii) a frameshift mutation, or iii) an acquired termination mutation (ie, a mutation that produces a premature stop codon (acquired stop codon )).

In some aspects, an effective amount of a chemotherapeutic agent is also administered as part of a treatment regimen.

In some embodiments, triacillib (on a regimen with specific timing) is received in combination with an effective amount of a PD-1 or PD-L1 checkpoint inhibitor and an effective amount of another immune checkpoint inhibitor (ICI) (selected from a group with Ig and ITIM domain T cell immune receptor (TIGIT) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint Inhibitors or cluster of differentiation 73 (CD73) checkpoint inhibitors) have CDK4/6-dependent NSCLC that 1) does not have at least one of the following: a. CCNE1 amplification; b. CCNE2 amplification; or c. Loss of retinoblastoma protein 1 (Rb1), which is defined as i) a homozygous deletion, ii) a frameshift mutation, or iii) an acquired termination mutation (ie, a mutation that produces a premature stop codon (acquired stop codon )); 2) have wild-type: i) CCNE1; ii) CCNE2; and iii) RB1; and 3) Have at least one of the following D cyclin activation features: i) CCND2 amplification; ii) CCND3 amplification; and iii) CCD1-3 3'UTR loss, defined as any of these UTRs Homozygous or heterozygous deletion.

In some aspects, an effective amount of a chemotherapeutic agent is also administered as part of a treatment regimen.

In some embodiments, triacillib (on a regimen with specific timing) is received in combination with an effective amount of a PD-1 or PD-L1 checkpoint inhibitor and an effective amount of another immune checkpoint inhibitor (ICI) (selected from a group with Ig and ITIM domain T cell immune receptor (TIGIT) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint Inhibitors or cluster of differentiation 73 (CD73) checkpoint inhibitors) in patients with CDK4/6-independent TNBC with at least one of the following characteristics: a. CCNE1 amplification; b. CCNE2 amplification; or c. Loss of retinoblastoma protein 1 (Rb1), which is defined as i) a homozygous deletion, ii) a frameshift mutation, or iii) an acquired termination mutation (ie, a mutation that produces a premature stop codon (acquired stop codon )).

In some aspects, an effective amount of a chemotherapeutic agent is also administered as part of a treatment regimen.

In some embodiments, triacillib (on a regimen with specific timing) is received in combination with an effective amount of a PD-1 or PD-L1 checkpoint inhibitor and an effective amount of another immune checkpoint inhibitor (ICI) (selected from a group with Ig and ITIM domain T cell immune receptor (TIGIT) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint Inhibitors or cluster of differentiation 73 (CD73) checkpoint inhibitors) have CDK4/6-dependent TNBC that 1) does not have at least one of the following: a. CCNE1 amplification; b. CCNE2 amplification; or c. Loss of retinoblastoma protein 1 (Rb1), which is defined as i) a homozygous deletion, ii) a frameshift mutation, or iii) an acquired termination mutation (ie, a mutation that produces a premature stop codon (acquired stop codon )); 2) have wild-type: i) CCNE1; ii) CCNE2; and iii) RB1; and 3) Have at least one of the following D cyclin activation features: i) CCND2 amplification; ii) CCND3 amplification; and iii) CCD1-3 3'UTR loss, defined as any of these UTRs Homozygous or heterozygous deletion.

In some aspects, an effective amount of a chemotherapeutic agent is also administered as part of a treatment regimen.

In some embodiments, triacillib (on a regimen with specific timing) is received in combination with an effective amount of a PD-1 or PD-L1 checkpoint inhibitor and an effective amount of another immune checkpoint inhibitor (ICI) (selected from a group with Ig and ITIM domain T cell immune receptor (TIGIT) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint Inhibitors or cluster of differentiation 73 (CD73) checkpoint inhibitors) in patients with CDK4/6-independent CRC with at least one of the following characteristics: a. CCNE1 amplification; b. CCNE2 amplification; or c. Loss of retinoblastoma protein 1 (Rb1), which is defined as i) a homozygous deletion, ii) a frameshift mutation, or iii) an acquired termination mutation (ie, a mutation that produces a premature stop codon (acquired stop codon )).

In some aspects, an effective amount of a chemotherapeutic agent is also administered as part of a treatment regimen.

In some embodiments, triacillib (on a regimen with specific timing) is received in combination with an effective amount of a PD-1 or PD-L1 checkpoint inhibitor and an effective amount of another immune checkpoint inhibitor (ICI) (selected from a group with Ig and ITIM domain T cell immune receptor (TIGIT) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint inhibitors or Cluster of Differentiation 73 (CD73) checkpoint inhibitors) in patients with CDK4/6-dependent CRC that: 1) Does not have at least one of the following items: a. CCNE1 amplification; b. CCNE2 amplification; or c. Loss of retinoblastoma protein 1 (Rb1), which is defined as i) a homozygous deletion, ii) a frameshift mutation, or iii) an acquired termination mutation (ie, a mutation that produces a premature stop codon (acquired stop codon )); 2) have wild-type: i) CCNE1; ii) CCNE2; and iii) RB1; and 3) Have at least one of the following D cyclin activation features: i) CCND2 amplification; ii) CCND3 amplification; and iii) CCD1-3 3'UTR loss, defined as any of these UTRs Homozygous or heterozygous deletion.

In some aspects, an effective amount of a chemotherapeutic agent is also administered as part of a treatment regimen.

In some embodiments, triacillib (on a regimen with specific timing) is received in combination with an effective amount of a PD-1 or PD-L1 checkpoint inhibitor and an effective amount of another immune checkpoint inhibitor (ICI) (selected from a group with Ig and ITIM domain T cell immune receptor (TIGIT) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint Inhibitors or cluster of differentiation 73 (CD73) checkpoint inhibitors) have CDK4/6-independent mUC with at least one of the following characteristics: a. CCNE1 amplification; b. CCNE2 amplification; or c. Loss of retinoblastoma protein 1 (Rb1), which is defined as i) a homozygous deletion, ii) a frameshift mutation, or iii) an acquired termination mutation (ie, a mutation that produces a premature stop codon (acquired stop codon )).

In some aspects, an effective amount of a chemotherapeutic agent is also administered as part of a treatment regimen.

In some embodiments, triacillib (on a regimen with specific timing) is received in combination with an effective amount of a PD-1 or PD-L1 checkpoint inhibitor and an effective amount of another immune checkpoint inhibitor (ICI) (selected from a group with Ig and ITIM domain T cell immune receptor (TIGIT) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint Inhibitors or cluster of differentiation 73 (CD73) checkpoint inhibitors) have CDK4/6-dependent mUC that: 1) Does not have at least one of the following items: a. CCNE1 amplification; b. CCNE2 amplification; or c. Loss of retinoblastoma protein 1 (Rb1), which is defined as i) a homozygous deletion, ii) a frameshift mutation, or iii) an acquired termination mutation (ie, a mutation that produces a premature stop codon (acquired stop codon )); 2) have wild-type: i) CCNE1; ii) CCNE2; and iii) RB1; and 3) Have at least one of the following D cyclin activation features: i) CCND2 amplification; ii) CCND3 amplification; and iii) CCD1-3 3'UTR loss, defined as any of these UTRs Homozygous or heterozygous deletion.

In some aspects, an effective amount of a chemotherapeutic agent is also administered as part of a treatment regimen.

In some embodiments, triacillib (on a regimen with specific timing) is received in combination with an effective amount of a PD-1 or PD-L1 checkpoint inhibitor and an effective amount of another immune checkpoint inhibitor (ICI) (selected from a group with Ig and ITIM domain T cell immune receptor (TIGIT) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint Inhibitors or cluster of differentiation 73 (CD73) checkpoint inhibitors) in patients with CDK4/6-independent solid tumors with at least one of the following characteristics: a. CCNE1 amplification; b. CCNE2 amplification; or c. Loss of retinoblastoma protein 1 (Rb1), which is defined as i) a homozygous deletion, ii) a frameshift mutation, or iii) an acquired termination mutation (ie, a mutation that produces a premature stop codon (acquired stop codon )).

In some aspects, an effective amount of a chemotherapeutic agent is also administered as part of a treatment regimen.

In some embodiments, triacillib (on a regimen with specific timing) is received in combination with an effective amount of a PD-1 or PD-L1 checkpoint inhibitor and an effective amount of another immune checkpoint inhibitor (ICI) (selected from a group with Ig and ITIM domain T cell immune receptor (TIGIT) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint inhibitors or Cluster of Differentiation 73 (CD73) checkpoint inhibitors) in patients with CDK4/6-dependent solid tumors that: 1) Does not have at least one of the following items: a. CCNE1 amplification; b. CCNE2 amplification; or c. Loss of retinoblastoma protein 1 (Rb1), which is defined as i) a homozygous deletion, ii) a frameshift mutation, or iii) an acquired termination mutation (ie, a mutation that produces a premature stop codon (acquired stop codon )); 2) have wild-type: i) CCNE1; ii) CCNE2; and iii) RB1; and 3) Have at least one of the following D cyclin activation features: i) CCND2 amplification; ii) CCND3 amplification; and iii) CCD1-3 3'UTR loss, defined as any of these UTRs Homozygous or heterozygous deletion.

In some aspects, an effective amount of a chemotherapeutic agent is also administered as part of a treatment regimen.

In some embodiments, the solid tumor patient treated with the specified timed regimen has a confirmed CDK4/6 independent solid tumor selected from the group consisting of: cervical cancer, squamous cell carcinoma of the head and neck (SCCHN), skin Squamous cell carcinoma (cSCC), Merkel cell carcinoma, basal cell carcinoma, small cell lung cancer (SCLC), melanoma, malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, high microsatellite instability or mismatch Repair-deficient cancer, endometrial cancer, high tumor mutation burden (TMB-H) cancer, gastric cancer, gastroesophageal junction cancer, esophageal adenocarcinoma, or esophageal cancer.

In some embodiments, the solid tumor patient treated with the specified timed regimen has a confirmed CDK4/6 dependent solid tumor selected from the group consisting of: cervical cancer, squamous cell carcinoma of the head and neck (SCCHN), skin Squamous cell carcinoma (cSCC), Merkel cell carcinoma, basal cell carcinoma, small cell lung cancer (SCLC), melanoma, malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, high microsatellite instability or mismatch Repair-deficient cancer, endometrial cancer, high tumor mutation burden (TMB-H) cancer, gastric cancer, gastroesophageal junction cancer, esophageal adenocarcinoma, or esophageal cancer.

Previous Immune Checkpoint Inhibitor Therapy Exposure In certain aspects of the present invention, provided herein are methods of treating patients with advanced/metastatic non-small cell lung cancer (NSCLC), advanced/metastatic III Negative breast cancer (TNBC), advanced/metastatic and unresectable colorectal cancer (CRC), advanced/metastatic urothelial carcinoma (mUC), or advanced/metastatic other solid tumors (such as head and neck squamous cell carcinoma) (SCCHN), cutaneous squamous cell carcinoma (cSCC), Merkel cell carcinoma, basal cell carcinoma, small cell lung cancer (SCLC), melanoma, malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, high microsatellite stable or mismatch repair-deficient cancer, endometrial cancer, high tumor mutation burden (TMB-H) cancer, gastric cancer, gastroesophageal junction (GEJ), esophageal adenocarcinoma, or esophageal cancer) who have previously Exposure to immune checkpoint inhibitors, such as inhibitors of programmed cell death protein-1 (PD-1) and/or programmed death-ligand-1 (PD-L1), in the first-line setting as monotherapy or in NSCLC and TNBC in combination with first-line chemotherapy regimens or alternatively in the setting of first-line chemotherapy in the case of CRC, mUC and other solid tumors) and have become therapeutically resistant to immune checkpoint inhibitors (thus eliciting after an initial response Disease progression). By using a regimen comprising administering a PD-1 or PD-L1 inhibitor and further comprising administering another ICI selected from a TIGIT inhibitor, a TIM-3 inhibitor, a LAG-3 inhibitor or a CD73 inhibitor (using or Administration of the short-acting, selective and reversible cyclin-dependent kinase (CDK) 4/6 inhibitor, treracilib, without the use of chemotherapy) overcomes the potential for dysregulation of cytotoxic T cell activity within the tumor microenvironment (TME) Mechanisms of immune checkpoint inhibitor resistance, thereby increasing the limited treatment options for these patients in the second- and third-line settings and increasing overall survival in a difficult-to-treat subgroup of patients.

Examples of immune checkpoint inhibitors and immunomodulators include, but are not limited to, PD-1 inhibitors, PD-L1 inhibitors, PD-L2 inhibitors, CTLA-4 inhibitors, LAG-3 inhibitors, TIM-3 Inhibitors, CD73 inhibitors, V domain Ig inhibitors of T cell activation (VISTA) inhibitors, TIGIT inhibitors, Siglec-15 inhibitors, B7-H3 (CD272) inhibitors, BTLA inhibitors (CD272), small molecule, peptide, nucleotide or another inhibitor. In certain aspects, the immunomodulator is an antibody, such as a monoclonal antibody.

In some embodiments, a PD-1 inhibitor has been previously administered to the patient. PD-1 inhibitors for use in the methods described herein include, for example but not limited to, nivolumab (OPDIVO ^® ), pembrolizumab (KEYTRUDA ^® ), simiprizumab (LIBTAYO ^® ; Regeneron), dotalimab (JEMPERLI ^® ), pilizumab (Medivation), AMP-224 (AstraZeneca/Medimmune), AMP-514 (AstraZeneca), sintilimab (IBI308; Innovent/Eli Lilly), saxenlimab (PF-06801591; Pfizer), sparizumab (PDR001; Novartis), rivelizumab (MGA012/INCMGA00012; Incyte Corporation and MacroGenics), tislelizumab (BGB -A317; BeiGene), toripalimab (JS001), camrelizumab (SHR-1210; Jiangsu Hengrui Medicine Company and Incyte Corporation), CS1003 (Cstone Pharmaceuticals), zimbelizumab (AB122; Arcus Biosciences) and JTX-4014 (Jounce Therapeutics).

In some embodiments, the patient has been previously administered a PD-L1 inhibitor. PD-L1 inhibitors for use in the methods described herein include, for example but not limited to, atezolizumab (TECENTRIQ ^® , Genentech), durvalumab (IMFINZI ^® , AstraZeneca), avelumab (BAVENCIO ^® ; Merck), Envolizumab (KN035; Alphamab), BMS-936559 (Bristol-Myers Squibb), BMS-986189 (Bristol-Myers Squibb), Lodalizumab (LY3300054; Eli Lilly), Ke Xili Monoclonal antibody (CK-301; Checkpoint Therapeutics), sugemalimumab (CS-1001; Cstone Pharmaceuticals), adelbelimumab (SHR-1316; Jiangsu HengRui Medicine), CBT-502 (CBT Pharma), AUNP12 (Aurigene), CA-170 (Aurigene/Curis) and BGB-A333 (BeiGene).

In some embodiments, the dual PD-L1/PD-1 inhibitor has been previously administered to the patient.

In some embodiments, a PD-L1/VISTA inhibitor has been previously administered to the patient. PD-L1-VISTA inhibitors include, but are not limited to, CA-170 (Curis Inc.). In some embodiments, the immune checkpoint inhibitor is a VISTA immune checkpoint inhibitor. VISTA inhibitors include, but are not limited to, JNJ-61610588 (Johnson & Johnson).

In some embodiments, a CTLA-4 immune checkpoint inhibitor has been previously administered to the patient. CTLA-4 inhibitors include, but are not limited to, ipilimumab (YERVOY ^® , Bristol Myers Squibb), tremelimumab (AstraZeneca/MedImmune), zalifrelimab (AGEN1884; Agenus ) and AGEN2041 (Agenus).

In some embodiments, the patient has previously been administered a LAG-3 immune checkpoint inhibitor. LAG-3 inhibitors for use in the methods described herein include, for example but not limited to, relarimab (OPDUALAG ^® ; BMS-986016; Bristol-Myers Squibb), GSK2831781 (GlaxoSmithKline), Effiragimo De alpha (IMP321; Prima BioMed), liramumab (LAG525; Novartis), favrizumab (MK-4280; Merck), framilizumab (REGN3767; Regeneron), TSR-033 (Tesaro/ GSK), BI754111 (Boehringer Ingelheim), Sym022 (Symphogen), LBL-007 (Nanjing Leads Biolabs Co., Ltd), IBI110 (Innovent Biologics), IBI323 (Innovent Biologics), INCAGN02385 (Incyte Corporation), AVA021 (A vacta), MGD013 (Macrogenics), RO7247669 (Hoffman-LaRoche), EMB-02 (Shanghai Epimab Biotherapeutics), XmAb841 (Xencor), dual PD-1 and LAG-3 inhibitor tepolizumab (MGD013; MacroGenics), CB213 (Crescendo Biologics) and SNA-03 (Microbio Group) and the dual PD-L1 and LAG-3 inhibitor FS118 (F-Star).

In some embodiments, the TIM-3 immune checkpoint inhibitor has been previously administered to the patient. TIM-3 inhibitors for use in the methods described herein include, for example but not limited to, cobrelimab (TSR-022; Tesaro), RG7769 (Genentech), MAS825 (Novartis), sabatolimab (MBG453; Novartis), Sym023 (Symphogen), INCAGN2390 (Incyte), LY3321367 (Eli Lilly and Company), BMS-986258 (BMS), SHR-1702 (Jiangsu HengRui), AZD7789 (AstraZeneca), TQB2618 (Chia Tai T ianqing Pharmaceutical Group Co., Ltd.) and NB002 (Neologics Bioscience), BGBA425 (Beigene) and Tim-3 and PD-1 bispecific RO7121661 (Roche).

In some embodiments, a TIGIT (T cell immune receptor with Ig and ITIM domains) immune checkpoint inhibitor has been previously administered to the patient. TIGIT (T cell immunoreceptor with Ig and ITIM domains) inhibitors for use in the methods described herein include, for example but not limited to, vebolizumab (MK-7684; Merck), etekimab /OMP-313 M32 (OncoMed), Tireliumab (MTIG7192A/RG-6058; Roche/Genentech), Ospolizumab (BGB-A1217; Beigene), BMS-986207 (BMS), COM902 ( Compugen), M6223 (Merck KGaA), dovanumab (AB-154; Arcus Biosciences), AZD2936 (AstraZeneca), JS006 (Shanghai Junshi Bioscience), IBI139 (Innovent Biologics), ASP-8374 (Astellas/Potenza), BAT6021 (Bio-Thera Solutions), TAB006 (Shanghai Junshi Bioscience), Domvanalimab (AB154; Arcus Biosciences), EOS884448 (EOS-448; iTeos), SEA-TGT (Seattle Genetics), mAb-7 (Stanwei Biotech), SHR- 1708 (Hengrui Medicine), GS02 (Suzhou Zelgen/Qilu Pharma), RXI-804 (Rxi Pharmaceuticals), NB6253 (Northern Biologics), ENUM009 (Enumreal Biomedical), CASC-674 (Cascadian Therapeutics), AJUD008 (AJUD Bioph arma), AGEN1777 (Agenus, Bristol-Myers Squibb), HLX53 (Shanghai Henlius Biotech), BAT6005 (Bio-Thera Solutions), anti-TIGIT/anti-PD-L1 bispecific antibody HLX301 (Shanghai Henlius Biotech), anti-TIGIT/anti-PD-L1 antibody HB0036 (Shanghai Huaota Biopharmaceutical).

In some embodiments, a CD73 (ecto-5' nucleotidase; (NT5E) checkpoint inhibitor has been previously administered to the patient. CD73 checkpoint inhibitors for use in the methods described herein include, for example but not Limited to) HLX23 (Shanghai Henlius Biotech), LY3475070 (Eli Lilly and Company), IPH5301 (Innate Pharma, Astra Zeneca), AK119 (Akesobio Australia Pty Ltd.), PT199 (Phanes Therapeutics), Mupadolizumab (CPI-006 ; Corvus Pharmaceuticals), Sym024 (Symphogen), Olevolumab (MEDI9447; Astra Zeneca), IBI325 (Innovent Biologics), ORIC-533 (Oric Pharmaceuticals), JAB-BX102 (Jacobio Pharmaceuticals), TJ004309 (Tracon Pharmaceuticals) icals), AB680 (Arcus Biosciences), NZV930 (Novartis), BMS-986179 (Bristol Myers Squibb), INCA00186 (Incyte Corporation) and anti-CD73-TGFβ-Trap bifunctional antibody Darovup α (Gilead Sciences).

In some embodiments, the patient has previously been administered an immune checkpoint inhibitor including, for example but not limited to, a B7-H3/CD276 immune checkpoint inhibitor (e.g., enotuzumab ( enoblituzumab) (MGA217, Macrogenics), MGD009 (Macrogenics), 131I-8H9/omburtamab (Y-mabs) and I-8H9/omburtamab (Y-mabs)), indoleamine 2, 3-Dioxygenase (IDO) immune checkpoint inhibitors (such as Indoximod (Indoximod) and INCB024360), killer immunoglobulin-like receptor (KIR) immune checkpoint inhibitors (such as lililumab ( Lirilumab) (BMS-986015)), carcinoembryonic antigen cell adhesion molecule (CEACAM) inhibitors (such as CEACAM-1, -3 and/or -5). Exemplary anti-CEACAM-1 antibodies are described in WO 2010/125571, WO 2013/082366, and WO 2014/022332, such as monoclonal antibodies 34B1, 26H7, and 5F4; or recombinant forms thereof (e.g., US 2004/0047858, US Patent No. 7,132,255 and described in WO 99/052552). In other embodiments, the anti-CEACAM antibody binds to CEACAM-5, as described in, for example, Zheng et al., PLoS One. 2010 Sep 2;5(9).pii:e12529 (DOI: 10:1371/journal. pone.0021146); or cross-reactive with CEACAM-1 and CEACAM-5, as described, for example, in WO 2013/054331 and US 2014/0271618.

In some embodiments, the patient has previously been administered an immune checkpoint inhibitor against CD47 including, but not limited to, Hu5F9-G4 (Stanford University/Forty7), TI-061 (Arch Oncology) , TTI-622 (Trillum Therapeutics), TTI-621 (Trillum Therapeutics), SRF231 (Surface Oncology), SHR-1603 (Hengrui), OSE-172 (Boehringer Ingelheim/OSE Immunotherapeutics), NI-1701 (Novimmune TG Therapeutics), IBI188 (Innovent Biologics), CC-95251 (Celgene), CC-90002 (Celgene/Inibrx), AO-176 (Arch Oncology), ALX148 (ALX Oncology), IMM01 (ImmuneOnco Biopharma), IMM2504 (ImmuneOnco Biopharma), IMM25 02 ( ImmuneOnco Biopharma), IMM03 (ImmuneOnco Biopharma), IMC-002 (ImmuneOncia Therapeutics), IBI322 (Innovent Biologics), HMBD-004B (Hummingbird Bioscience), HMBD-004A (Hummingbird Bioscience), HLX24 (Henlius), FSI -189 (Forty Seven), DSP107 (KAHR Medical), CTX-5861 (Compass Therapeutics), BAT6004 (Bio-Thera), AUR-105 (Aurigene), AUR-104 (Aurigene), Anti-CD47 (Biocad), ABP-500 (Abpro) , ABP-160 (Abpro), TJC4 (I-MAB Biopharma), TJC4-CK (I-MAB Biopharma), SY102 (Saiyuan), SL-172154 (Shaattuck Labs), PSTx-23 (Paradigm Shift Therapeutics), PDL1/ CD47BsAb (Hanmi Pharmaceuticals), NI-1801 (Novimmune), MBT-001 (Morphiex), LYN00301 (LynkCell) and BH-29xx (Beijing Hanmi).

In some embodiments, the patient has previously been administered an immune checkpoint inhibitor against CD39, including but not limited to TTX-030 (Tizona Therapeutics), IPH5201 (Innate Pharma/AstraZeneca), SRF -617 (Surface Oncology), ES002 (Elpisciences), 9-8B (Igenica) and antisense oligonucleotides (Secarna).

In some embodiments, the patient has previously been administered an immune checkpoint inhibitor against B and T lymphocyte attenuating molecule (BTLA), e.g., as described in Zhang et al., Monoclonal antibodies to B and T lymphocyte attenuator (BTLA) have no effect on in vitro B cell proliferation and act to inhibit in vitro T cell proliferation when presented in a cis, but not trans, format relative to the activating stimulus, Clin Exp Immunol. 2011 Jan; 163(1): 77-87 and TAB004/JS004 (Junshi Biosciences).

In some embodiments, the patient has previously been administered a sialic acid binding immunoglobulin-like lectin 15 (Siglec-15) inhibitor including, but not limited to, NC318 (anti-Siglec-15 mAb).

Modified regimen As contemplated herein, administration of the short-acting, selective and reversible cyclin-dependent kinase (CDK) 4/6 inhibitor treracilil or a pharmaceutically acceptable salt thereof (on a specified timed regimen) Check with PD-1 or PD-L1 checkpoint inhibitors and checkpoint inhibitors selected from T cell immune receptors with Ig and ITIM domains (TIGIT), T cell immunoglobulin mucin-3 (TIM-3) Other immune checkpoint inhibitors (ICIs) that are point inhibitors, lymphocyte activation gene 3 (LAG-3) checkpoint inhibitors, or CD73 inhibitors. In some aspects, chemotherapeutic agents are also administered as part of the treatment regimen. In certain embodiments, administration of the selective CDK4/6 inhibitor triraciclib in combination with a PD-1 or PD-L1 inhibitor and another ICI as described herein increases the potency of immune checkpoint inhibition (including overcoming resistance to development of resistance to previously administered PD-1 or PD-L1 inhibitors and/or reducing or delaying the onset of resistance), thereby prolonging the efficacy of anti-cancer regimens. In certain embodiments, administration of the selective CDK4/6 inhibitor triraciclib together with PD-1 or PD-L1 inhibitors and other ICIs as described herein can improve survival outcomes for these refractory patients ( Including overall survival (OS) and/or progression-free survival (PFS)) and reducing or delaying resistance to ICI therapy. Specific cancers amenable to treatment using the methods of the present invention include, but are not limited to, non-small cell lung cancer (NSCLC), advanced/metastatic triple-negative breast cancer (TNBC), advanced/metastatic and unresectable colorectal cancer (CRC) , advanced/metastatic urothelial carcinoma (mUC) and other solid tumors (including (but not limited to) cervical cancer, squamous cell carcinoma of the head and neck (SCCHN), squamous cell carcinoma of the skin (cSCC), Merkel cell carcinoma , basal cell carcinoma, small cell lung cancer (SCLC), melanoma, malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, microsatellite instability-high or mismatch repair-deficient cancers, endometrial cancer, high tumor mutation burden (TMB-H) cancer, gastric cancer, gastroesophageal junction cancer, esophageal adenocarcinoma, and esophageal cancer).

Triracilil (2'-((5-(4-methylhexahydropyrazin-1-yl)pyridin-2-yl)amino)-7',8'-dihydro-6'H -Spiro(cyclohexane-1,9'-pyrazino(1',2':1,5)pyrrolo(2,3-d)pyrimidin)-6'-one) is highly selective with the structure Sexual CDK4/6 inhibitors: .

Triracillib, or a pharmaceutically acceptable salt, composition, isotopic analog or prodrug thereof, is administered using a suitable carrier as provided herein. Triracilib is commercially available as ^COSELA® (G1 Therapeutics, Inc.). Traxilib is described in US 2013-0237544, the entire contents of which case is incorporated herein by reference. Triracillib can be synthesized as described in US 2019-0135820, the entire contents of which are incorporated herein by reference. Triracillib can be administered in any manner that achieves the desired result, including systemically, parenterally, intravenously, intramuscularly, subcutaneously, or intradermally. For injection, sterile, lyophilized, yellowish Trirasili is served in pie form. The product, for example, can be supplied in single-use 20-mL clear glass vials and is preservative-free. For example, 19.5 ml of 0.9% Sodium Chloride Injection or 5% Dextrose Injection may be used to reconstitute Treracilil for Injection (300 mg/vial) prior to administration. This reconstituted solution has a concentration of straciclib of 15 mg/mL and is usually subsequently diluted prior to intravenous administration. As described herein, treracilil can be administered intravenously.

In certain embodiments, treracicill is in the dihydrochloride form and optionally in the hydrate form. For example, treracillib can be used in the present invention in the form of dihydrochloride dihydrochloride or in the form of a pharmaceutical composition formed from triracillib dihydrochloride dihydrate.

In some embodiments, treracilil is administered at about 180 mg/m ² to 300 mg/m ² . In some embodiments, at about 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, or about 280 mg/m ² cast treracilide. In some embodiments, treracicill is administered at at least 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, or 240 mg/ ^m2 . In some embodiments, about 4 hours or less prior to the administration of the chemotherapeutic agent (e.g., about 4 hours or less, 3 hours or less prior to the administration of the chemotherapeutic agent or the first chemotherapeutic agent to be administered in a combination regimen, respectively). 1 hour or less, 2 hours or less, about 1 hour or less, or about 30 minutes), treracicill is administered at about 240 mg/m ² . In some embodiments, treracillib is administered intravenously over a period of about 30 minutes. In some embodiments, the treracilib is administered completely prior to the administration.

In an alternative embodiment, a different CDK4/6 inhibitor is administered. For example, in alternative embodiments, the CDK4/6 inhibitor used in place of treraciclib in the regimens described herein is ribociclib (Novartis), palbociclib (Pfizer), or Abemaciclib (Eli Lilly) or a pharmaceutically acceptable salt thereof. In other alternative embodiments, the CDK4/6 inhibitor is lerociclib having the following structure:

,

or a pharmaceutically acceptable composition, salt, isotope analog or prodrug thereof, which is described in US 2013-0237544 (the entire content of which is incorporated herein by reference), and can be obtained as described in US 2019-0135820 (the entirety of which The contents of which are incorporated herein by reference) were synthesized as described. In some embodiments, lerosilicate is administered as a pharmaceutically acceptable salt (eg, dihydrochloride).

In other alternative embodiments, the CDK4/6 inhibitor has the following structure: , or a pharmaceutically acceptable composition, salt, isotope analog or prodrug thereof, which is described in US 2013-0237544 (the entire content of which is incorporated herein by reference), and can be described in US 2019-0135820 (which The entire content is incorporated herein by reference) as described in Synthesis.

In an alternative embodiment, instead of treracilib, a CDK4/6 inhibitor selected from palbociclib, ribociclib or abeciclib is used.

PD-1 Inhibitor In one embodiment, the first immune checkpoint inhibitor is a PD-1 inhibitor that blocks the interaction of PD-1 and PD-L1 by binding to the PD-1 receptor and then Suppression of immunosuppression. In one embodiment, the immune checkpoint inhibitor is selected from the following PD-1 immune checkpoint inhibitors: nivolumab (OPDIVO ^® ), pembrolizumab (KEYTRUDA ^® ), cimipril monoclonal antibody Anti-(LIBTAYO ^® ; Regeneron), dotalimab (JEMPERLI ^® ), pilizumab (Medivation), AMP-224 (AstraZeneca/Medimmune), AMP-514 (AstraZeneca), sintilimab (IBI308 ; Innovent/Eli Lilly), saxolizumab (PF-06801591; Pfizer), sparizumab (PDR001; Novartis), rivelizumab (MGA012/INCMGA00012; Incyte Corporation and MacroGenics), tislelizumab Monoclonal antibody (BGB-A317; BeiGene), toripalizumab (JS001), camrelizumab (SHR-1210; Jiangsu Hengrui Medicine Company and Incyte Corporation), CS1003 (Cstone Pharmaceuticals), zimbelizumab anti (AB122; Arcus Biosciences) and JTX-4014 (Jounce Therapeutics).

In one embodiment, the PD-1 inhibitor is nivolumab (OPDIVO ^® ), administered in an effective amount for the treatment of unresectable or metastatic melanoma, early and metastatic non-small cell lung cancer (NSCLC), intermediate or low risk advanced renal cell carcinoma (RCC), recurrent or progressive classical Hodgkin lymphoma (Hodgkin lymphoma), recurrent or metastatic squamous cell carcinoma of the head and neck (SCCHN), locally advanced Metastatic or metastatic urothelial carcinoma (mUC), microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR) metastatic colorectal cancer (CRC), hepatocellular carcinoma, unresectable malignant pleura Mesothelioma, adjuvant or metastatic esophageal cancer, gastroesophageal junction (GEJ) cancer, and gastric cancer. In one embodiment, nivolumab is administered at 240 mg every 2 weeks or at 480 mg every 4 weeks for NSCLC, CRC and mUC. In one embodiment, nivolumab is administered prior to chemotherapy.

In some embodiments, the PD-1 inhibitor is pembrolizumab (KEYTRUDA ^® ), administered in an effective amount for the treatment of unresectable or metastatic melanoma, metastatic non-small cell lung cancer (NSCLC) , intermediate or high risk advanced renal cell carcinoma (RCC), relapsed or refractory classical Hodgkin's lymphoma, recurrent or metastatic squamous cell carcinoma of the head and neck (SCCHN), locally advanced or metastatic urothelial carcinoma (mUC), high tumor mutation burden (TMB-H) cancer, locally recurrent unresectable or metastatic triple-negative breast cancer (TNBC), recurrent or metastatic cutaneous squamous cell carcinoma (cSCC), high microsatellite Stable (MSI-H) or mismatch repair deficient (dMMR) cancer, microsatellite instability high (MSI-H) or mismatch repair deficient (dMMR) metastatic colorectal cancer (CRC), hepatocellular carcinoma , adjuvant or metastatic esophageal cancer, gastroesophageal junction (GEJ) cancer, advanced endometrial cancer, recurrent locally advanced or metastatic Merkel cell carcinoma, locally advanced unresectable or metastatic gastric cancer, recurrent Acute or metastatic cervical cancer or primary mediastinal large B-cell lymphoma (PMBCL). In one embodiment, pembrolizumab is administered at 200 mg every 3 weeks or at 400 mg every 6 weeks for NSCLC, TNBC, CRC and mUC. In one embodiment, pembrolizumab is administered prior to chemotherapy.

In one embodiment, the PD-1 inhibitor is simiprimumab (LIBTAYO ^® ), administered in an effective amount for the treatment of locally advanced or metastatic cutaneous squamous cell carcinoma (CSCC), locally advanced Metastatic or metastatic non-small cell lung cancer (NSCLC), locally advanced or metastatic basal cell carcinoma, metastatic triple-negative breast cancer (TNBC), metastatic colorectal cancer (CRC), or metastatic urothelial carcinoma (mUC). In one embodiment, cimiprizumab 350 mg is administered as an intravenous infusion over 30 minutes every 3 weeks until disease progression. In one embodiment, cimiprizumab is administered prior to chemotherapy.

In one embodiment, the PD-1 inhibitor is dotalimab (JEMPERLI ^® ), administered in an effective amount for the treatment of mismatch repair deficient (dMMR) recurrent or advanced endometrial cancer, in Progressed solid tumors, metastatic non-small cell lung cancer (NSCLC), metastatic triple-negative breast cancer (TNBC), metastatic colorectal cancer (CRC) or metastatic urothelial carcinoma (mUC) without alternative treatment options. In one embodiment, dotalimab 500 mg is administered as an intravenous infusion over 30 minutes every 3 weeks until disease progression. In one embodiment, dotalimab is administered prior to chemotherapy.

PD-L1 Inhibitor In one embodiment, the first immune checkpoint inhibitor is a PD-L1 inhibitor that blocks the interaction of PD-1 and PD-L1 by binding to the PD-L1 receptor and then Suppression of immunosuppression. PD-L1 inhibitors include atezolizumab (TECENTRIQ ^® , Genentech), durvalumab (IMFINZI ^® , AstraZeneca), avelumab (BAVENCIO ^® ; Merck), envolimumab (KN035; Alphamab), BMS-936559 (Bristol-Myers Squibb), BMS-986189 (Bristol-Myers Squibb), lodalizumab (LY3300054; Eli Lilly), coxilimab (CK-301; Checkpoint Therapeutics), sugemalimumab Anti-(CS-1001; Cstone Pharmaceuticals), Adebelizumab (SHR-1316; Jiangsu HengRui Medicine), CBT-502 (CBT Pharma), AUNP12 (Aurigene), CA-170 (Aurigene/Curis) and BGB- A333 (BeiGene).

In one embodiment, the immune checkpoint inhibitor is the PD-L1 immune checkpoint inhibitor atezolizumab (TECENTRIQ ^® ), administered in an effective amount for the treatment of locally advanced or metastatic urothelial carcinoma (mUC), unresectable or metastatic melanoma, metastatic triple-negative breast cancer (TNBC), metastatic colorectal cancer (CRC), metastatic non-small cell lung cancer (NSCLC), unresectable or metastatic hepatocellular carcinoma or metastatic small cell lung cancer (SCLC). In one embodiment, atezolizumab is administered at 840 mg every 2 weeks, 1200 mg every 3 weeks, or 1680 mg every 4 weeks. In one embodiment, atezolizumab is administered prior to chemotherapy.

In another aspect of this embodiment, the immune checkpoint inhibitor is the PD-L1 immune checkpoint inhibitor durvalumab (IMFINZI ^® ), administered in an effective amount for the treatment of non-small cell lung cancer ( NSCLC), extensive-stage small cell lung cancer (SCLC), metastatic triple-negative breast cancer (TNBC), metastatic colorectal cancer (CRC), or metastatic urothelial carcinoma (mUC). In one embodiment, administered at 10 mg/kg every 2 weeks or at 1500 mg every 4 weeks (for patients weighing greater than 30 kg) and at 10 mg/kg every 2 weeks (for patients weighing less than 30 kg) with durvalumab. In one embodiment, durvalumab is administered prior to chemotherapy.

In another aspect of this embodiment, the immune checkpoint inhibitor is the PD-L1 immune checkpoint inhibitor avelumab (BAVENCIO ^® ), administered in an effective amount for the treatment of Merkel cell carcinoma, Metastatic urothelial carcinoma (mUC), renal cell carcinoma (RCC), metastatic triple-negative breast cancer (TNBC), metastatic colorectal cancer (CRC), or metastatic urothelial carcinoma (mUC). In one embodiment, avelumab is administered at 800 mg every 2 weeks. In one embodiment, avelumab is administered prior to chemotherapy.

with immunoglobulin and ITIM domain of T cellular immune receptor (TIGIT) Inhibitor

The T cell immune receptor with immunoglobulin and ITIM domains (TIGIT) is a promising new target for cancer immunotherapy. TIGIT is upregulated by immune cells including activated T cells, natural killer cells and regulatory T cells. TIGIT binds to two ligands, CD155 (PVR) and CD112 (PVRL2, cohesin-2), which are expressed by tumor cells and antigen-presenting cells in the tumor microenvironment (Stanietsky et al., The interaction of TIGIT with PVR and PVRL2 inhibits human NK cell cytotoxicity. Proc Natl Acad Sci U S A 2009; 106: 17858-63).

TIGIT (also known as WUCAM, Vstm3, VSIG9) is an Ig superfamily receptor that plays a key role in limiting adaptive and innate immunity (Boles et al., A novel molecular interaction for the adhesion of follicular CD4 T cells to follicular DC. Eur J Immunol 2009; 39:695-703). TIGIT involvement involves multiple inhibitory receptors (e.g. CD96/TACTILE, CD112R/PVRIG), a competitive co-stimulatory receptor (DNAM-1/CD226), and various ligands (e.g. CD155 (PVR/NECL-5), CD112 (mucosin, The complex regulatory network of zonulin-2/PVRL2) (Levin et al., Vstm3 is a member of the CD28 family and an important modulator of T-cell function. Eur J Immunol 2011; 41: 902-15; Bottino et al., Identification of PVR (CD155) and nectin-2 (CD112) as cell surface ligands for the human DNAM-1 (CD226) activating molecule. J Exp Med 2003; 198: 557-67; Seth et al., The murine pan T cell marker CD96 is an adhesion receptor for CD155 and nectin-1. Biochem Biophys Res Commun 2007; 364: 959-65; Zhu et al., Identification of CD112R as a novel checkpoint for human T cells. J Exp Med 2016; 213: 167-76 ).

TIGIT is expressed in humans by activated CD8+ T and CD4+ T cells, natural killer (NK) cells, regulatory T cells (Treg) and follicular T helper cells (Joller et al., Cutting edge: TIGIT has T cell-intrinsic inhibitory functions. J Immunol 2011; 186: 1338-42; Wu et al., Follicular regulatory T cells repress cytokine production by follicular helper T cells and optimize IgG responses in mice. Eur J Immunol 2016; 46: 1152-61). In marked contrast to DNAM-1/CD226, TIGIT is weakly expressed by naive T cells. In cancer, TIGIT is coexpressed with PD-1 on tumor antigen-specific CD8+ T cells and CD8+ tumor infiltrating lymphocytes (TILs) in mice and humans (Chauvin et al., TIGIT and PD-1 impairment tumor antigen- specific CD8⁺ T cells in melanoma patients. J Clin Invest 2015; 125: 2046-58; Johnston et al., The immunoreceptor TIGIT regulates antitumor and antiviral CD8(+) T cell effector function. Cancer Cell 2014; 26 :923-37 ) . It is also coexpressed with other inhibitory receptors such as T cell immunoglobulin and mucin domain-containing molecule-3 (TIM-3) and lymphocyte activation gene 3 (LAG-3) in depleted CD8+ T cells in tumors. On cell subgroups (Chauvin et al., TIGIT and PD-1 impairment tumor antigen-specific CD8⁺ T cells in melanoma patients. J Clin Invest 2015; 125: 2046-58; Johnston et al., The immunoreceptor TIGIT regulates antitumor and antiviral CD8 (+) T cell effector function. Cancer Cell 2014; 26 :923-37). In addition, TIGIT is highly expressed by T _reg in peripheral blood mononuclear cells of healthy donors and cancer patients and further upregulated in TME (Joller et al., Treg cells expressing the coinhibitory molecule TIGIT selectively inhibit proinflammatory Th1 and Th17 cell responses. Immunity 2014; 40: 569-81; Zhang et al., Genome-Wide DNA methylation analysis identifies hypomethylated genes regulated by FOXP3 in human regulatory T cells. Blood 2013; 122: 2823-36).

In one embodiment, the other immune checkpoint inhibitor is a TIGIT inhibitor, which blocks the interaction of TIGIT and CD155 by binding to the TIGIT receptor and then inhibits immunosuppression. TIGIT inhibitors include vibralimab (MK-7684; Merck), etiriliumab/OMP-313 M32 (OncoMed), tisrelumab (MTIG7192A/RG-6058; Roche/Genentech), Perlimumab (BGB-A1217; Beigene), BMS-986207 (BMS), COM902 (Compugen), M6223 (Merck KGaA), dorvanumab (AB-154; Arcus Biosciences), AZD2936 (AstraZeneca), JS006 (Shanghai Junshi Bioscience), IBI139 (Innovent Biologics), ASP-8374 (Astellas/Potenza), BAT6021 (Bio-Thera Solutions), TAB006 (Shanghai Junshi Bioscience), Domvanalimab (AB154; Arcus Biosciences), EOS884448 (EOS-448; iTeos), SEA-TGT (Seattle Genetics), mAb-7 (Stanwei Biotech), SHR-1708 (Hengrui Medicine), GS02 (Suzhou Zelgen/Qilu Pharma), RXI-804 (Rxi Pharmaceuticals), NB6253 (Northern Biologics), ENUM009 (Enumreal Biomedical), CASC-674 (Cascadian Therapeutics), AJUD008 (AJUD Biopharma), AGEN1777 (Agenus, Bristol-Myers Squibb), HLX53 (Shanghai Henlius Biotech), BAT6005 (Bio-Thera Solutions), anti-TIGIT/anti-PD -L1 bispecific antibody HLX301 (Shanghai Henlius Biotech), anti-TIGIT/anti-PD-L1 antibody HB0036 (Shanghai Huaota Biopharmaceutical).

In one embodiment, a TIGIT inhibitor is used in combination with a CDK4/6 inhibitor and a PD-1 inhibitor. In one embodiment, a TIGIT inhibitor is used in combination with a CDK4/6 inhibitor, a PD-1 inhibitor and chemotherapy. In one embodiment, the CDK4/6 inhibitor is treracicil. In one embodiment, the PD-1 inhibitor is Nivolumab. In one embodiment, the PD-1 inhibitor is pembrolizumab. In one embodiment, the PD-1 inhibitor is simiprizumab. In one embodiment, the PD-1 inhibitor is dotalimab. In one embodiment, the PD-1 inhibitor blocks the interaction between PD-1 and PD-L1 to inhibit immunosuppression. In one embodiment, a TIGIT inhibitor blocks the interaction between TIGIT and CD155 to inhibit immunosuppression. In one embodiment, the combination of a CDK4/6 inhibitor, a PD-1 inhibitor, a TIGIT inhibitor, and chemotherapy produces greater tumor suppression than a combination without a CDK4/6 inhibitor and a TIGIT inhibitor. In one embodiment, a CDK4/6 inhibitor, a PD-1 or PD-L1 inhibitor, and a TIGIT inhibitor are not used with chemotherapy.

In one embodiment, a TIGIT inhibitor is used in combination with a CDK4/6 inhibitor. In one embodiment, a TIGIT inhibitor is used in combination with a CDK4/6 inhibitor and a PD-L1 inhibitor. In one embodiment, a TIGIT inhibitor is used in combination with a CDK4/6 inhibitor, a PD-L1 inhibitor and chemotherapy. In one embodiment, the CDK4/6 inhibitor is treracicil. In one embodiment, the PD-L1 inhibitor is atezolizumab. In one embodiment, the PD-L1 inhibitor is avelumab. In one embodiment, the PD-L1 inhibitor is durvalumab. In one embodiment, the PD-L1 inhibitor blocks the interaction between PD-L1 and CD80 to inhibit immunosuppression. In one embodiment, a TIGIT inhibitor blocks the interaction between TIGIT and CD155 to inhibit immunosuppression. In one embodiment, the combination of a CDK4/6 inhibitor, a PD-L1 inhibitor, a TIGIT inhibitor, and chemotherapy produces greater tumor suppression than a combination without a CDK4/6 inhibitor and a TIGIT inhibitor. In one embodiment, CDK4/6 inhibitors, PD-L1 inhibitors and TIGIT inhibitors are not used with chemotherapy.

T cell immunoglobulin and mucin domain 3 (TIM-3) inhibitor T cell immunoglobulin and mucin domain 3 (TIM-3) (encoded by Havcr2 ) contains immunoglobulin (Ig) and mucin A cell surface molecule of the T domain, which was originally identified as a cell surface marker specific for interferon (IFN-γ)-producing CD4 ⁺ T helper 1 (Th1) and CD8 ⁺ T cytotoxicity 1 (Tc1) cells found (Monney et al., Th1-specific cell surface protein TIM-3 regulates macrophage activation and severity of an autoimmune disease. Nature 2002; 415: 536-41). Tim-3 is co-regulated with other immune checkpoint receptors (PD-1, Lag-3 and TIGIT) and co-expressed on CD4 ⁺ and CD8 ⁺ T cells (Chihara et al., Induction and transcriptional regulation of the co-inhibitory gene module in T cells. Nature 2018; 558: 454-9; DeLong et al., Il-27 and TCR stimulation promote T cell expression of multiple inhibitory receptors. ImmunoHorizons 2019; 3: 13-25). In cancer, TIM-3 expression specifically marks the most dysfunctional or terminally exhausted subset of CD8 ⁺ T cells (Fourcade et al., Upregulation of TIM-3 and PD-1 expression is associated with tumor antigen-specific CD8+ T cell dysfunction in melanoma patients. J Exp Med 2010; 207: 2175-86; Sakuishi et al., Targeting TIM-3 and PD-1 pathways to reverse T cell exhaustion and restore anti-tumor immunity. J Exp Med 2010; 207: 2187 -94). The following four TIM-3 ligands have been identified: galectin-9, phosphatidylserine (PtdSer), high mobility group box B1 (HMGB1 ), and CEACAM-1.

In one embodiment, the other immune checkpoint inhibitor is a TIM-3 inhibitor, which blocks TIM-3 and galectin-9, phosphatidylserine (PtdSerine) by binding to the TIM-3 receptor ), high mobility group box B1 (HMGB1 ) and/or CEACAM-1 interaction and subsequent suppression of immunosuppression. TIM-3 inhibitors include coprimab (TSR-022; Tesaro), RG7769 (Genentech), MAS825 (Novartis), sabatolimab (MBG453; Novartis), Sym023 (Symphogen), INCAGN2390 (Incyte) , LY3321367 (Eli Lilly and Company), BMS-986258 (BMS), SHR-1702 (Jiangsu HengRui), AZD7789 (AstraZeneca), TQB2618 (Chia Tai Tianqing Pharmaceutical Group Co., Ltd.) and NB002 (Neologics Bioscience), BGBA 425 (Beigene) and TIM-3 and PD-1 bispecific RO7121661 (Roche).

In one embodiment, a TIM-3 inhibitor is used in combination with a CDK4/6 inhibitor. In one embodiment, a TIM-3 inhibitor is used in combination with a CDK4/6 inhibitor and a PD-1 inhibitor. In one embodiment, TIM-3 inhibitors are used in combination with CDK4/6 inhibitors, PD-1 inhibitors and chemotherapy. In one embodiment, the CDK4/6 inhibitor is treracicil. In one embodiment, the PD-1 inhibitor is Nivolumab. In one embodiment, the PD-1 inhibitor is pembrolizumab. In one embodiment, the PD-1 inhibitor is simiprizumab. In one embodiment, the PD-1 inhibitor is dotalimab. In one embodiment, the PD-1 inhibitor blocks the interaction between PD-1 and PD-L1 to inhibit immunosuppression. In one embodiment, the TIM-3 inhibitor blocks the interaction between TIM-3 and galectin-9, phosphatidylserine (PtdSer), high mobility group box B1 (HMGB1) and/or CEACAM-1 interaction to suppress immunosuppression. In one embodiment, the combination of a CDK4/6 inhibitor, a PD-1 inhibitor, a TIM-3 inhibitor, and chemotherapy produces greater tumor suppression than a combination without a CDK4/6 inhibitor and a TIM-3 inhibitor. In one embodiment, CDK4/6 inhibitors, PD-1 inhibitors and TIM-3 inhibitors are not used with chemotherapy.

In one embodiment, a TIM-3 inhibitor is used in combination with a CDK4/6 inhibitor. In one embodiment, a TIM-3 inhibitor is used in combination with a CDK4/6 inhibitor and a PD-L1 inhibitor. In one embodiment, a TIM-3 inhibitor is used in combination with a CDK4/6 inhibitor, a PD-L1 inhibitor and chemotherapy. In one embodiment, the CDK4/6 inhibitor is treracicil. In one embodiment, the PD-L1 inhibitor is atezolizumab. In one embodiment, the PD-L1 inhibitor is durvalumab. In one embodiment, the PD-L1 inhibitor is avelumab. In one embodiment, the PD-L1 inhibitor blocks the interaction between PD-L1 and CD80 to inhibit immunosuppression. In one embodiment, the TIM-3 inhibitor blocks the interaction between TIM-3 and galectin-9, phosphatidylserine (PtdSer), high mobility group box B1 (HMGB1) and/or CEACAM-1 interaction to suppress immunosuppression. In one embodiment, the combination of a CDK4/6 inhibitor, a PD-L1 inhibitor, a TIM-3 inhibitor, and chemotherapy produces greater tumor suppression than a combination without a CDK4/6 inhibitor and a TIM-3 inhibitor. In one embodiment, CDK4/6 inhibitors, PD-L1 inhibitors and TIM-3 inhibitors are not used with chemotherapy.

The lymphocyte activation gene -3 (LAG-3) inhibitor LAG‐3 (CD223) is encoded by the LAG‐3 gene. LAG‐3 is a member of the immunoglobulin superfamily (IgSF) and exerts numerous biological influences on T cell function (Triebel et al., LAG‐3, a novel lymphocyte activation gene closely related to CD4. J Exp Med 1990; 171: 1393 -405). LAG‐3 is expressed on the membranes of natural killer cells (NK), B cells, tumor infiltrating lymphocytes (TIL, a subset of T cells), and dendritic cells (DC) (Triebel et al., LAG‐3, a novel lymphocyte activation gene closely related to CD4. J Exp Med 1990; 171: 1393-405); Kisielow et al., Expression of lymphocyte activation gene 3 (LAG‐3) on B cells is induced by T cells. Eur J Immunol 2005; 35 : 2081-8; Grosso et al., LAG‐3 regulates CD8+ T cell accumulation and effector function in murine self‐ and tumor‐tolerance systems. J Clin Invest 2007; 117: 3383-92; Workman et al., LAG‐3 regulates plasmacytoid dendritic cell homeostasis. J Immunol 2009; 182: 1885-91; Andreae et al., Maturation and activation of dendritic cells induced by lymphocyte activation gene‐3 (CD223). J Immunol 2002; 168: 3874-80). The LAG-3 protein binds the non-holotropic region of the major histocompatibility complex 2 (MHC class II) with an affinity greater than that of CD 4 (Baixeras et al., Characterization of the lymphocyte activation gene 3-encoded protein. A new ligand for human leukocyte antigen class II antigens. J Exp Med 1992; 176: 327-37). LAG‐3 is one of several immune checkpoint receptors that are synergistically upregulated on regulatory T cells (T _reg ) and anergic T cells, and blockade Discontinuation enhances the reversal of this anergic state (Grosso et al., Functionally distinct LAG-3 and PD-1 subsets on activated and chronically stimulated CD8 T cells. J Immunol 2009; 182: 6659-69). LAG‐3/MHC class II interaction downregulates CD4+ Ag-specific T cell clone responses following LAG‐3 binding. Eur J Immunol 1996; 26: 1180-6).

In one embodiment, the other immune checkpoint inhibitor is a LAG-3 inhibitor, which blocks the interaction between LAG-3 and major histocompatibility complex 2 (MHC class II) by binding to the LAG-3 receptor. Interact and in turn inhibit immunosuppression. LAG-3 inhibitors include, but are not ^limited to, relagimod (IMP321; Ramirizumab (LAG525; Novartis), Favizlizumab (MK-4280; Merck), Framilizumab (REGN3767; Regeneron), TSR-033 (Tesaro/GSK), BI754111 (Boehringer Ingelheim), Sym022 (Symphogen), LBL-007 (Nanjing Leads Biolabs Co., Ltd), IBI110 (Innovent Biologics), IBI323 (Innovent Biologics), INCAGN02385 (Incyte Corporation), AVA021 (Avacta), MGD013 (Macrogenics), RO7247669 (Hoffman -La Roche ), EMB-02 (Shanghai Epimab Biotherapeutics), XmAb841 (Xencor), dual PD-1 and LAG-3 inhibitor tepolizumab (MGD013; MacroGenics), CB213 (Crescendo Biologics) and SNA-03 (Microbio Group) And dual PD-L1 and LAG-3 inhibitor FS118 (F-Star).

In one embodiment, a LAG-3 inhibitor is used in combination with a CDK4/6 inhibitor. In one embodiment, a LAG-3 inhibitor is used in combination with a CDK4/6 inhibitor and a PD-1 inhibitor. In one embodiment, a LAG-3 inhibitor is used in combination with a CDK4/6 inhibitor, a PD-1 inhibitor and chemotherapy. In one embodiment, the CDK4/6 inhibitor is treracicil. In one embodiment, the PD-1 inhibitor is Nivolumab. In one embodiment, the PD-1 inhibitor is pembrolizumab. In one embodiment, the PD-1 inhibitor is simiprizumab. In one embodiment, the PD-1 inhibitor is dotalimab. In one embodiment, the LAG-3 inhibitor is Rilarimab. In one embodiment, the PD-1 inhibitor blocks the interaction between PD-1 and PD-L1 to inhibit immunosuppression. In one embodiment, a LAG-3 inhibitor blocks the interaction of LAG-3 with major histocompatibility complex 2 (MHC class II) by binding to the LAG-3 receptor and thereby inhibits immunosuppression. In one embodiment, the combination of a CDK4/6 inhibitor, a PD-1 inhibitor, a LAG-3 inhibitor, and chemotherapy produces greater tumor suppression than a combination without a CDK4/6 inhibitor and a LAG-3 inhibitor. In one embodiment, CDK4/6 inhibitors, PD-1 inhibitors and LAG-3 inhibitors are not used with chemotherapy.

In one embodiment, a LAG-3 inhibitor is used in combination with a CDK4/6 inhibitor. In one embodiment, a LAG-3 inhibitor is used in combination with a CDK4/6 inhibitor and a PD-L1 inhibitor. In one embodiment, a LAG-3 inhibitor is used in combination with a CDK4/6 inhibitor, a PD-L1 inhibitor and chemotherapy. In one embodiment, the CDK4/6 inhibitor is treracicil. In one embodiment, the PD-L1 inhibitor is atezolizumab. In one embodiment, the PD-L1 inhibitor is durvalumab. In one embodiment, the PD-L1 inhibitor is avelumab. In one embodiment, the LAG-3 inhibitor is Rilarimab. In one embodiment, the PD-L1 inhibitor blocks the interaction between PD-L1 and CD80 to inhibit immunosuppression. In one embodiment, a LAG-3 inhibitor blocks the interaction of LAG-3 with major histocompatibility complex 2 (MHC class II) by binding to the LAG-3 receptor and thereby inhibits immunosuppression. In one embodiment, the combination of a CDK4/6 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, and chemotherapy produces greater tumor suppression than a combination without a CDK4/6 inhibitor and a LAG-3 inhibitor. In one embodiment, CDK4/6 inhibitors, PD-L1 inhibitors and LAG-3 inhibitors are not used with chemotherapy.

Exo -5'- Nucleotidase / Cluster of Differentiation 73 (NT5E; CD73) Inhibitor ), which is an enzyme encoded by the NT5E gene in humans (Misumi Y et al., (August 1990). "Primary structure of human placental 5'-nucleotidase and identification of the glycolipid anchor in the mature form". European Journal of Biochemistry. 191(3): 563-9). CD73 is commonly used to convert AMP to adenosine (Allard et al., "Chapter Fifteen - Measurement of CD73 enzymatic activity using luminescence-based and colorimetric assays", Methods in Enzymology, Tumor Immunology and Immunotherapy - Molecular Methods, Academic Press, 62 9: 269-289). CD73 is a membrane-bound extracellular enzyme that is overexpressed in several types of cancer. Its manifestations are associated with poorer prognosis in melanoma, colorectal cancer, gastric cancer, triple-negative breast cancer, and a pro-metastatic phenotype in prostate cancer (Stagg J et al., CD73-deficient mice are resistant to carcinomaogenesis. Cancer Res. 2012 5 Jan 1;72(9):2190-6). Together with CD39, it plays a major role in promoting immunosuppression via the pathway that degrades adenosine triphosphate (ATP) to adenosine. Within the tumor microenvironment, ATP promotes immune cell-mediated cancer cell killing. In contrast, adenosine accumulation induces immunosuppression, dysregulation of immune cell infiltrates, and stimulation of angiogenesis, leading to tumor dissemination. CD73 is active in the final step of the degradation pathway, where it is the enzyme that degrades AMP to adenosine. CD73 blockade promotes antitumor immunity by reducing adenosine accumulation. Thus, anti-CD73 mAb stimulates anti-tumor immunity and reduces tumor metastasis in mouse tumor models, and can enhance the therapeutic efficacy of anti-PD1 or anti-CTLA4 antibodies (Allard et al., Targeting CD73 enhances the antitumor activity of anti-PD- 1 and anti-CTLA-4 mAbs. Clin Cancer Res. 2013 Oct 15;19(20):5626-35). Figure 19 shows a visual representation of the effects of adenosinergic molecules on tumors and surrounding stroma (Vijayan D et al., Targeting immunosuppressive adenosine in cancer. Nat Rev Cancer. 2017 Dec;17(12):709-724). Adenosine is a widely described immunosuppressant that attenuates the effector functions of various immune cell populations, including T cells, and enhances the suppressive functions of T regs. Adenosine accumulation benefits tumor growth and metastasis through effects on tumor cells and stroma. For example, activation of CD73 on tumor cells may benefit cell adhesion and inhibit tumor cell apoptosis via epidermal growth factor receptor (EGFR) signaling (indicated by dashed arrows). CD73 activation also releases matrix metalloproteinases (MMPs) that facilitate the breakdown of extracellular matrix (ECM), thereby enabling tumor cells to invade and migrate to distant organs. In addition, A2BR activation on tumor cells promotes proliferation and angiogenesis through secretion of vascular endothelial growth factor (VEGF). In cancer-associated fibroblasts (CAFs), A2BR induces fibroblast growth factor 2 (FGF2) expression and increases the number of fibroblast activation protein (FAP)-positive fibroblasts. A2BR activation leads to increased release of _CXCL12 from these FAP+ fibroblasts, thereby increasing the number of intratumoral CD31+ endothelial cells. A2BR can engage the G protein-coupled receptor Gq protein kinase C (PKC) signaling pathway to activate interleukin-6 (IL-6), which in turn mediates epithelial-mesenchymal transition (EMT). In addition, some tumor exosomes co-express CD39 and CD73, which may be related to the spread of tumors to distant organs (Vijayan, D., Young, A., Teng, MWL, & Smyth, MJ (2017). Targeting immunosuppressive adenosine in cancer. Nature Reviews Cancer, 17(12), 709-724).

In one embodiment, the other immune checkpoint inhibitor is a CD73 inhibitor that specifically binds to CD73 and blocks its extracellular 5'-nucleotidase activity. By reducing the production of adenosine, CD73 inhibitors can alleviate the inhibitory effect of adenosine on the proliferation and tumor killing activity of CD8+ T cells, and weaken the stimulation of adenosine on immunosuppressive cells, thereby regulating the tumor microenvironment and enhancing anti-tumor immunity reaction. CD73 inhibitors include (but are not limited to) HLX23 (Shanghai Henlius Biotech), LY3475070 (Eli Lilly and Company), IPH5301 (Innate Pharma, Astra Zeneca), AK119 (Akesobio Australia Pty Ltd.), PT199 (Phanes Therapeutics), Mupa Dolimumab (CPI-006; Corvus Pharmaceuticals), Sym024 (Symphogen), Olevolumab (MEDI9447; Astra Zeneca), IBI325 (Innovent Biologics), ORIC-533 (Oric Pharmaceuticals), JAB-BX102 (Jacobio Pharmaceuticals) , TJ004309 (Tracon Pharmaceuticals), AB680 (Arcus Biosciences), NZV930 (Novartis), BMS-986179 (Bristol Myers Squibb), INCA00186 (Incyte Corporation) and anti-CD73-TGFβ-Trap bifunctional antibody Darovup α (Gilead Sciences).

In one embodiment, a CD73 inhibitor is used in combination with a CDK4/6 inhibitor. In one embodiment, a CD73 inhibitor is used in combination with a CDK4/6 inhibitor and a PD-1 inhibitor. In one embodiment, a CD73 inhibitor is used in combination with a CDK4/6 inhibitor, a PD-1 inhibitor and chemotherapy. In one embodiment, the CDK4/6 inhibitor is treracicil. In one embodiment, the PD-1 inhibitor is Nivolumab. In one embodiment, the PD-1 inhibitor is pembrolizumab. In one embodiment, the PD-1 inhibitor is simiprizumab. In one embodiment, the PD-1 inhibitor is dotalimab. In one embodiment, the PD-1 inhibitor blocks the interaction between PD-1 and PD-L1 to inhibit immunosuppression. In one embodiment, the CD73 inhibitor specifically binds to CD73 and blocks its extracellular 5'-nucleotidase activity. In one embodiment, the combination of a CDK4/6 inhibitor, a PD-1 inhibitor, a CD73 inhibitor, and chemotherapy produces greater tumor suppression than a combination without a CDK4/6 inhibitor and a CD73 inhibitor. In one embodiment, CDK4/6 inhibitors, PD-1 inhibitors and CD73 inhibitors are not used with chemotherapy.

In one embodiment, a CD73 inhibitor is used in combination with a CDK4/6 inhibitor. In one embodiment, a CD73 inhibitor is used in combination with a CDK4/6 inhibitor and a PD-L1 inhibitor. In one embodiment, a CD73 inhibitor is used in combination with a CDK4/6 inhibitor, a PD-L1 inhibitor and chemotherapy. In one embodiment, the CDK4/6 inhibitor is treracicil. In one embodiment, the PD-L1 inhibitor is atezolizumab. In one embodiment, the PD-L1 inhibitor is durvalumab. In one embodiment, the PD-L1 inhibitor is avelumab. In one embodiment, the PD-L1 inhibitor blocks the interaction between PD-L1 and CD80 to inhibit immunosuppression. In one embodiment, the CD73 inhibitor specifically binds to CD73 and blocks its extracellular 5'-nucleotidase activity. In one embodiment, the combination of a CDK4/6 inhibitor, a PD-L1 inhibitor, a CD73 inhibitor, and chemotherapy produces greater tumor suppression than a combination without a CDK4/6 inhibitor and a CD73 inhibitor. In one embodiment, CDK4/6 inhibitors, PD-L1 inhibitors and CD73 inhibitors are not used with chemotherapy.

Other Immune Checkpoint Inhibitors In some embodiments, a surrogate immune checkpoint inhibitor is administered to the patient instead of administering the TIGIT, LAG-3, TIM-3, or CD73 checkpoint inhibitor to the patient. In some embodiments, the surrogate immune checkpoint inhibitor is a B7-H3/CD276 immune checkpoint inhibitor (e.g., enotuzumab (MGA217, Macrogenics) MGD009 (Macrogenics), 131I-8H9/omatuzumab ( Y-mabs) and I-8H9/omatumab (Y-mabs)), indoleamine 2,3-dioxygenase (IDO) immune checkpoint inhibitors (such as Insimod and INCB024360), killer Immunoglobulin-like receptor (KIR) immune checkpoint inhibitors (such as lililumab (BMS-986015)), carcinoembryonic antigen cell adhesion molecule (CEACAM) inhibitors (such as CEACAM-1, -3 and/or -5). Exemplary anti-CEACAM-1 antibodies are described in WO 2010/125571, WO 2013/082366, and WO 2014/022332, such as monoclonal antibodies 34B1, 26H7, and 5F4; or recombinant forms thereof (e.g., US 2004/0047858, US Patent No. 7,132,255 and described in WO 99/052552). In other embodiments, the anti-CEACAM antibody binds to CEACAM-5 as, for example, Zheng et al., PLoS One. 2010 Sep 2; 5(9). pii: e12529 (DOI: 10: 1371/journal. pone.0021146); or cross-reactive with CEACAM-1 and CEACAM-5, as described, for example, in WO 2013/054331 and US 2014/0271618.

In some embodiments, the alternative immune checkpoint inhibitor is an inhibitor against CD47, including but not limited to Hu5F9-G4 (Stanford University/Forty7), TI-061 (Arch Oncology), TTI-622 (Trillum Therapeutics) , TTI-621 (Trillum Therapeutics), SRF231 (Surface Oncology), SHR-1603 (Hengrui), OSE-172 (Boehringer Ingelheim/OSE Immunotherapeutics), NI-1701 (Novimmune TG Therapeutics), IBI188 (Innovent Biologics), CC- 95251 (Celgene), CC-90002 (Celgene/Inibrx), AO-176 (Arch Oncology), ALX148 (ALX Oncology), IMM01 (ImmuneOnco Biopharma), IMM2504 (ImmuneOnco Biopharma), IMM2502 (ImmuneOnco Biopharma), IMM M03 (ImmuneOnco Biopharma ), IMC-002 (ImmuneOncia Therapeutics), IBI322 (Innovent Biologics), HMBD-004B (Hummingbird Bioscience), HMBD-004A (Hummingbird Bioscience), HLX24 (Henlius), FSI-189 (Forty Seven), DSP107 (KAHR Medical) , CTX-5861 (Compass Therapeutics), BAT6004 (Bio-Thera), AUR-105 (Aurigene), AUR-104 (Aurigene), Anti-CD47 (Biocad), ABP-500 (Abpro), ABP-160 (Abpro), TJC4 (I-MAB Biopharma), TJC4-CK (I-MAB Biopharma), SY102 (Saiyuan), SL-172154 (Shaattuck Labs), PSTx-23 (Paradigm Shift Therapeutics), PDL1/ CD47BsAb (Hanmi Pharmaceuticals), NI- 1801 (Novimmune), MBT-001 (Morphiex), LYN00301 (LynkCell) and BH-29xx (Beijing Hanmi).

In some embodiments, the alternative immune checkpoint inhibitor is an inhibitor against CD39, including but not limited to TTX-030 (Tizona Therapeutics), IPH5201 (Innate Pharma/AstraZeneca), SRF-617 (Surface Oncology), ES002 (Elpisciences), 9-8B (Igenica) and antisense oligonucleotides (Secarna).

In some embodiments, the alternative immune checkpoint inhibitor is an inhibitor against B and T lymphocyte attenuator (BTLA), e.g., as described in Zhang et al., Monoclonal antibodies to B and T lymphocyte attenuator (BTLA) have no effect on in In vitro B cell proliferation and act to inhibit in vitro T cell proliferation when presented in a cis, but not trans, format relative to the activating stimulus, Clin Exp Immunol. 2011 Jan; 163(1): 77-87 and TAB004/JS004 (Junshi Biosciences).

In some embodiments, the surrogate immune checkpoint inhibitor is a sialic acid binding immunoglobulin-like lectin 15 (Siglec-15) inhibitor, including but not limited to NC318 (anti-Siglec-15 mAb).

Chemotherapeutic agents may be administered in combination with treracilil, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors, and CD73 inhibitors and Any standard chemotherapeutic agent treatment modality.

In one embodiment, the chemotherapeutic agent is toxic to immune effector cells. In one embodiment, the chemotherapeutic agent inhibits cell growth. In one embodiment, the cytotoxic chemotherapeutic agent administered is a DNA damaging chemotherapeutic agent. In one embodiment, the chemotherapeutic agent is a protein synthesis inhibitor, a DNA damaging chemotherapeutic agent, an alkylating agent, a topoisomerase inhibitor, an RNA synthesis inhibitor, a DNA complex binding agent, a thiolate alkyl Anticancer agents, guanine alkylating agents, tubulin binding agents, DNA polymerase inhibitors, anticancer enzymes, RAC1 inhibitors, thymidylate synthase inhibitors, oxazaphosphine compounds, integrins Inhibitors (eg cilengitide, camptothecin or homocamptothecin), antifolates or folate antimetabolites.

Cytotoxic Chemotherapeutics Cytotoxic, DNA damaging chemotherapeutics tend to be nonspecific and toxic to normal, rapidly dividing cells such as HSPCs and immune effector cells, especially at high doses. As used herein, the term "DNA damaging" chemotherapy or chemotherapeutic agent refers to the use of cytostatic or cytotoxic agents (i.e., compounds) to reduce or eliminate the growth or Treatment of proliferation, where the cytotoxic effect of the agent can be the result of one or more of the following: nucleic acid intercalation or conjugation, DNA or RNA alkylation, inhibition of RNA or DNA synthesis, inhibition of another nucleic acid-associated activity (e.g. protein synthesis), or any other Cytotoxic effect. Such compounds include, but are not limited to, DNA damaging compounds that can kill cells. "DNA damaging" chemotherapeutic agents include, but are not limited to, alkylating agents, DNA intercalators, protein synthesis inhibitors, DNA or RNA synthesis inhibitors, DNA base analogs, topoisomerase inhibitors, telomere Enzyme inhibitors and telomeric DNA binding compounds. Alkylating agents include, for example, alkylsulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodizepa; ), carboquone, meturedepa and uredepa; ethyleneimine and methylmelamine such as altretamine, triethylenemelamine, triethylene Nitrogen mustards such as chlorambucil, chlornaphazine, cyclophosphamide, Estramustine, Mechlorethamine, Mechlorethamine Oxide Hydrochloride, Melphalan, Novembichine, Cholesterol Phenesterine, Prednimol prednimustine, trofosfamide, and uracil mustard; and nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine and ranimustine. Other DNA damaging chemotherapeutic agents include daunorubicin, doxorubicin, idarubicin, epirubicin, mitomycin, and streptozocin. Chemotherapeutic antimetabolites include gemcitabine, mercaptopurine, thioguanine, cladribine, fludarabine phosphate, 5-FU, floxuridine, arabinosin Cytarabine, pentostatin, methotrexate, azathioprine, acyclovir, adenine β-1-D-arabinoside, methotrexate ( Amethopterin), aminopterin (aminopterin), 2-aminopurine, aphidicolin, 8-azaguanine, azaserine, 6-azauracil, 2′-azides Nitro-2′-deoxynucleoside, 5-bromodeoxycytidine, cytosine β-1-D-arabinoside, diazooxynorleucine, dideoxynucleoside, 5-fluoro Deoxycytidine, 5-fluorodeoxyuridine and hydroxyurea.

Chemotherapeutic protein synthesis inhibitors include abrin, aurintricarboxylic acid, chloramphenicol, colicin E3, cycloheximide, diphtheria toxin, edeine A, ipecacin ( emetine), erythromycin, ethionine, fluoride, 5-fluorotryptophan, fusidic acid, guanosyl methylene diphosphonate and imido Guanosyl diphosphonate, kanamycin, kasugamycin, kirromycin and O-methylthreonine. Other protein synthesis inhibitors include modeccin, neomycin, norvaline, pactamycin, paromomycine, puromycin, castor Toxin, shiga toxin, showdomycin, sparsomycin, spectinomycin, streptomycin, tetracycline, thiostrepton (thiostrepton) and trimethoprim (trimethoprim).

DNA synthesis inhibitors include alkylating agents such as dimethyl sulfate, nitrogen and sulfur mustards; intercalating agents such as acridine dyes, actinomycin, anthracene, benzopyrene, ethidium bromide , propidium diiodide-intertwining; and other agents such as distamycin and netropsin. Topoisomerase inhibitors (eg, irinotecan, teniposide, coumermycin, nalidixic acid, novobiocin, and oxolinic acid) may also be used. acid)); cell division inhibitors (including colcemide, mitoxantrone, colchicine, vinblastine, and vincristine); and RNA synthesis inhibitors (including actinomycin D, α -Amanitine and other fungal toxins, cordycepin (3′-deoxyadenosine), dichlororibofuranosyl benzimidazole, rifampicine, Streptococcus varicosities streptovaricin and streptolydigin as DNA damaging compounds.

In one embodiment, the chemotherapeutic agent is a DNA complex binding agent such as camptothecin or etoposide; a thiolate alkylating agent such as nitrosourea, BCNU, CCNU, ACNU or formustine; Guanine alkylating agents such as temozolomide; tubulin binding agents such as vinblastine, vincristine, vinorelbine, vinflunine, cryptophycin 52, halichondrin (e.g. soft sponge B), dolastatin (such as dolastatin 10 and dolastatin 15), hemiasterlin (such as hemiasterlin A and hemiasterlin B), colchicum colchicine, combrestatin, 2-methoxyestradiol, E7010, paclitaxel, docetaxel, epothilone, discodermolide; DNA polymerase Inhibitors such as cytarabine; anticancer enzymes such as asparaginase; Rac1 inhibitors such as 6-thioguanine; thymidylate synthase inhibitors such as capecitabine or 5- FU; oxazaphosphorinane compounds such as Cytoxan; integrin inhibitors such as cilengitide; antifolates such as pralatrexate; folate antimetabolites , such as pemetrexed ; or camptothecin or homocamptothecin, such as diflomotecan.

In one embodiment, the topoisomerase inhibitor is a type I inhibitor. In another embodiment, the topoisomerase inhibitor is a type II inhibitor.

Other DNA damaging chemotherapeutics whose toxic effects can be mitigated by the selective CDK4/6 inhibitors disclosed herein include, but are not limited to, cisplatin, hydrogen peroxide, carboplatin, procarbazine, ifosf Amide, bleomycin, plicamycin, taxol, transplatinum, thiotepa, oxaliplatin and the like and similar acting agents. In one embodiment, the DNA damaging chemotherapeutic agent is selected from the group consisting of cisplatin, carboplatin, camptothecin and etoposide.

Other suitable chemotherapeutic agents include, but are not limited to, radioactive molecules, toxins (also known as cytotoxins or cytotoxic agents, which include any agent detrimental to cell viability), agents containing chemotherapeutic compounds, and liposomes or other vesicles Bubble. General anticancer drugs include: vincristine (ONCOVIN ^® ), liposomal vincristine (MARQIBO ^® ), doxorubicin (ADRIAMYCIN ^® ), cytarabine (cytosine arabinoside, ara-C or CYTOSAR ^® ), L-asparaginase (ELSPAR ^® ) or PEG-L-asparaginase (pegaspargase or ONCASPAR ^® ), etoposide (VP-16), teniposide Glycosides (VUMON ^® ), 6-mercaptopurine (6-MP or purine THOL ^® ), prednisone and dexamethasone (DECADRON ^® ). Examples of other suitable chemotherapeutic agents include, but are not limited to, 5-fluorouracil, dacarbazine, alkylating agents, anthramycin (AMC)), antimitotic agents, cis-dichlorodiamidoplatinum ( II) (DDP) cisplatin), diaminodichloroplatinum, anthracycline, antibiotic, antimetabolite, asparaginase, live BCG (intravesical), bleomycin sulfate, calicheamicin ), cytochalasin B, dactinomycin (formerly known as actinomycin), daunorubicin hydrochloride, daunorubicin citrate, denileukin diftitox, dihydroxy dihydroxy anthracin dione, docetaxel, doxorubicin hydrochloride, Escherichia coli (E. coli) L-asparaginase, Erwinia (Erwinia) L-asparagine Amidase, etoposide, citrovorum factor, etoposide phosphate, gemcitabine hydrochloride, idarubicin hydrochloride, interferon alpha-2b, irinotecan hydrochloride, maytansoid Maytansinoid, dichloroethylmethylamine hydrochloride, melphalan hydrochloride, mithramycin, mitomycin C, mitotane, paclitaxel, polyphenylpropanol ( polifeprosan 20 with carmustine implant, procarbazine hydrochloride, streptozotocin, teniposide, thiotepa, topotecan hydrochloride, valrubicin Star, vinblastine sulfate, vincristine sulfate and vinorelbine tartrate.

Other cytotoxic chemotherapeutic agents useful in the present invention include: epirubicin, abraxane, taxotere, epothilone, tafluposide, vimodegil ( vismodegib), azacitidine, doxifluridine, vindesine, and vinorelbine.

In one embodiment, the chemotherapeutic agent is a DNA complex binding agent. In one embodiment, the chemotherapeutic agent is a tubulin binding agent. In one embodiment, the chemotherapeutic agent is an alkylating agent. In one embodiment, the chemotherapeutic agent is a thiolate alkylating agent.

Other chemotherapeutics Other chemotherapeutics that can be used as described herein can include 2-methoxyestradiol or 2ME2, finasunate, etaracizumab (MEDI-522), HLL1, huN901-DM1, atiprimod, saquinavir mesylate, ritonavir, nelfinavir mesylate, indina sulfate Indinavir sulfate, plitidepsin, P276-00, tipifarnib, lenalidomide, thalidomide, pomalidomide, Simvastatin and celecoxib. Chemotherapeutic agents useful in the present invention include, but are not limited to, Trastuzumab (HERCEPTIN ^® ), Pertuzumab (PERJETA ^® ), Lapatinib (TYKERB ^® ), Gefitinib (IRESSA ^® ), Erlotinib (TARCEVA ^® ), Cetuximab (ERBITUX ^® ), Panitumumab (VECTIBIX ^® ), Vandetanib ( Vandetanib (CAPRELSA ^® ), Vemurafenib (ZELBORAF ^® ), Vorinostat (ZOLINZA ^® ), Romidepsin (ISTODAX ^® ), Bexarotene ( TARGRETIN ^® ), Alitretinoin (PANRETIN ^® ), Tretinoin (VESANOID ^® ), Carfilzomib (KYPROLIS ^® ), Pralatrexate (FOLOTYN ^® ), Bevacizate Zizumab (AVASTIN ^® ), Ziv-aflibercept (ZALTRAP ^® ), Sorafenib (NEXAVAR ^® ), Sunitinib (SUTENT ^® ), Pazopanib (Pazopanib) (VOTRIENT ^® ), Regorafenib (STIVARGA ^® ), and Cabozantinib (COMETRIQ ^® ).

Other chemotherapeutic agents contemplated include, but are not limited to, calcineurin inhibitors (e.g., cyclosporine or ascomycin, e.g., cyclosporine A ( ^NEORAL® ), FK506 (tacrolimus), pyridoxine pimecrolimus), mTOR inhibitors (such as rapamycin or its derivatives, such as Sirolimus (RAPAMUNE ^® ), Everolimus (CERTICAN ^® ), temsirolimus (temsirolimus, zotarolimus, bayerolimus-7, bayerolimus-9), rapamycin analogs (rapalog) (such as redaforolimus (ridaforolimus, campath 1H), S1P receptor modulators, dual mTORC1 and mTORC2 inhibitors (e.g. Vistusertib (AZD2014), e.g. fingolimod or its analogs ), anti-IL-8 antibody, mycophenolic acid or its salt (such as sodium salt) or its prodrug (such as Mycophenolate Mofetil (CELLCEPT ^® )), OKT3 (Orthoclone OKT3 ^® ), Thymoglobulin ^® , Brequinar Sodium, OKT4, T10B9.A- ^3A , 33B3.1, 15-deoxyspergualin, tresperimus , Leflunomide (ARAVA ^® ), anti-CD25, anti-IL2R, Basiliximab (SIMULECT ^® ), Daclizumab (ZENAPAX ^® ), mizoribine ), dexamethasone, ISAtx-247, SDZ ASM 981 (pimecrolimus, ELIDEL ^® ), abatacept, belatacept, LFA3lg, etanercept (with ^ENBREL® forms sold by ImmuneXcite), adalimumab ( ^HUMIRA® ), infliximab ( ^REMICADE® ), anti-LFA-1 antibody, natalizumab ( ^ANTEGREN® ), en Enlimomab, gavilimomab, Golimumab, anti-thymocyte immunoglobulin, siplizumab, alefacept, efalfabri Efalizumab, Pentasa, mesalazine, asacol, codeine phosphate, benorylate, fenbufen , naprosyn, diclofenac, etodolac, indomethacin, dasatinib (SPRYCEL ^® ), nilotinib (TASIGNA ^® ), bosutinib (BOSULIF ^® ), imatinib mesylate (GLEEVEC ^® ) and ponatinib (ICLUSIG ^® ), amifostine (amifostine), doral mesylate Dolasetron mesylate, dronabinol, epoetin-alpha, etidronate, filgrastim, fluconazole, acetic acid goserelin acetate, gramicidin D, granisetron, leucovorin calcium, lidocaine, mesna, ondan Setron (ondansetron) hydrochloride, pilocarpine (pilocarpine) hydrochloride, porfimer sodium (porfimer sodium), vatalanib (vatalanib), 1-testosterone, allopurinol sodium, betamethasone ( Betamethasone), sodium phosphate and betamethasone acetate, calcium folate, conjugated estrogens, dexrazoxane, dibromomannitol, esterified estrogens, estradiol, estramox Estramustine phosphate sodium, ethylene estradiol, flutamide, folinic acid, glucocorticoids, leuprolide acetate, levamisole hydrochloride, medroxyprogesterone acetate (medroxyprogesterone acetate), megestrol acetate, methyltestosterone, nilutamide, octreotide acetate, pamidronate disodium, progesterone Procaine, propranolol, testolactone, tetracaine, toremifene citrate, and sargramostim.

In one embodiment, the chemotherapeutic agent is an estrogen receptor ligand such as tamoxifen, raloxifene, fulvestrant, anordrin , bazedoxifene, broparestriol, chlorotrianisene, clomiphene citrate, cyclofenil, lasofoxifene, Ormeloxifene or toremifene; androgen receptor ligands such as bicalutamide, enzalutamide, apalutamide, cyclic acetate Cyproterone acetate, chlormadinone acetate, spironolactone, canrenone, drospirenone, ketoconazole, topilutamide ), abiraterone acetate, or cimetidine; aromatase inhibitors, such as letrozole, anastrozole, or exemestane; anti-inflammatory agents, For example, Presone; Oxidase inhibitors, such as allopurinol; Anti-cancer antibodies; Anti-cancer monoclonal antibodies; Antibodies against CD40, such as lucatumumab or dacetuzumab; CD20 antibodies, such as rituximab; CD52-binding antibodies, such as alemtuzumab; integrin-binding antibodies, such as volociximab or natalizumab; Antibodies against the IL-6 receptor, such as tocilizumab; interleukin-2 mimics, such as aldesleukin; antibodies targeting IGF1, such as figitumumab ); antibodies targeting DR4, such as mapatumumab; antibodies targeting TRAIL-R2, such as lexatumumab or dulanermin; fusion proteins, such as acetaminophen atacicept; B-cell inhibitors, such as acetacept; proteasome inhibitors, such as carfilzomib, bortezomib, or marizomib; HSP90 inhibitors, such as tanespimycin ); HDAC inhibitors such as vorinostat, belinostat, or panobinostat; MAPK ligands such as talmapimod; PKC inhibitors such as enzastaurin (enzastaurin); HER2 receptor ligands such as trastuzumab, lapatinib, or pertuzumab; EGFR inhibitors such as gefitinib, erlotinib, cetuximab, pertuzumab Nitumab or vandetanib; natural products such as romidepsin; retinoids such as bexarotene, tretinoin, or alitretinoin; receptor tyrosine kinase (RTK) inhibitors such as Vatinib, stretratinib, cabozantinib, sunitinib, regorafenib, or pazopanib; or VEGF inhibitors such as ziv-aflibercept, bevacizumab, or Dovitinib.

In one embodiment, CDK4/6 inhibitors, chemotherapeutics, and combinations of various immune checkpoint inhibitors are further used in combination with hematopoietic growth factors including, but not limited to, granulocyte colony-stimulating factor (G - CSF, e.g. sold as ^NEUPOGEN® (filgrastim), ^NEULASTA® (peg-filgrastim) or legrastim), granuloma-macrophage colony-stimulating factor (GM-CSF, e.g. Sold as molastim and sargragrastim (LEUKINE ^® ), M-CSF (macrophage colony stimulating factor), thrombopoietin (megakaryocyte growth and development factor (MGDF, e.g. as ROMIPLOSTIM ^® and ELTROMBOPAG ^® (sold in the form of ), interleukin (IL)-12, interleukin-3, interleukin-11 (adipogenic inhibitory factor or oprelin), SCF (stem cell factor, blue gray factor, kit-ligand or KL) and erythropoietin (EPO) and derivatives thereof (epoetin-alpha sold as DARBEPOEITIN ^® , EPOCEPT ^® , NANOKINE ^® , EPOFIT ^® , EPOGEN ^® , EPREX ^® and PROCRIT ^® ; ^® , RECORMON ^® and MICERA ^® ; Epoetine-δ (sold as e.g. DYNEPO ^® ), Epoetine-omega (sold as e.g. EPOMAX ^® ), Epoetine ζ ( sold as SILAPO ^® and RETACRIT ^® ) and such as EPOCEPT ^® , EPOTRUST ^® , ERYPRO ^® Safe, REPOITIN ^® , VINTOR ^® , EPOFIT ® , ERYKINE ^® , WEPOX ^® , ESPOGEN ^® , RELIPOEITIN ^® , SHANPOEITIN ^{® ,} ZYROP ^® , and ^EPIAO® ). In one embodiment, hematopoietic growth factors are administered timed such that the effect of the CDK4/6 inhibitor on HSPCs disappears. In one embodiment, the growth factor is administered at least 20 hours after administration of the CDK4/6 inhibitor. factor.

Other chemotherapeutic agents may include estrogen inhibitors including, but not limited to, SERMs (selective estrogen receptor modulators), SERDs (selective estrogen receptor degraders), complete estrogen receptor degraders, or partial Or another form of full estrogen antagonist. Some antiestrogens (such as raloxifene and tamoxifen) retain some estrogen-like effects, including estrogen-like uterine growth stimulation and (in some cases) estrogens that actually stimulate tumor growth during breast cancer progression same effect. In contrast, fulvestrant (a complete anti-estrogen) has no estrogen-like effects on the uterus and is effective in tamoxifen-resistant tumors. Non-limiting examples of antiestrogenic compounds are provided in WO 2014/19176 (assigned to Astra Zeneca), WO 2013/090921 , WO 2014/203129, WO 2014/203132 and US2013/0178445 (assigned to Olema Pharmaceuticals) and US Patent Nos. 9,078,871, 8,853,423 and 8,703,810 and in US 2015/0005286, WO 2014/205136 and WO 2014/205138. Other non-limiting examples of anti-estrogen compounds include: SERMs such as anordrin, bazedoxifene, bromotriol, clomiphene citrate, cyclofenyl, lasofoxifene, omexifen , raloxifene, tamoxifen, toremifene, and fulvestrant; aromatase inhibitors such as aminoglutethimide, testrolactone, anastrozole, exemestane, Fadrozole, formestane, and letrozole; and antigonadotropins such as leuprorelin, cetrorelix, allylestrenol, chlordiacetate Chloromadinone acetate, delmadinone acetate, dydrogesterone, medroxyprogesterone acetate, megestrol acetate, nomegestrol acetate, norethindrone acetate Ketone (norethisterone acetate), progesterone and spironolactone.

Other chemotherapeutic agents may include androgen (e.g., testosterone) inhibitors, including, but not limited to, selective androgen receptor modulators, selective androgen receptor degraders, complete androgen receptor degraders, or partial or complete Another form of androgen antagonist. In one embodiment, the prostate or testicular cancer is androgen resistant. Non-limiting examples of antiandrogenic compounds are provided in WO 2011/156518 and US Patent Nos. 8,455,534 and 8,299,112. Other non-limiting examples of antiandrogenic compounds include: chlormadinone acetate, spironolactone, canrenone, drospirenone, ketoconazole, topiramide, abiraterone acetate, and ximedidine.

Chemotherapeutic agents may comprise kinase inhibitors including, but not limited to, phosphoinositide 3-kinase (PI3K) inhibitors, Bruton's tyrosine kinase (BTK) inhibitors, or spleen tyrosine kinase (Syk) inhibitors or combinations thereof.

PI3k inhibitors are well known. Examples of PI3 kinase inhibitors include, but are not limited to, Wortmannin, demethoxyviridin, perifosine, idelalisib, picoxib ( pictilisib), Palomid 529, ZSTK474, PWT33597, CUDC-907 and AEZS-136, duvelisib, GS-9820, GDC-0032 (2-[4-[2-(2-isopropyl-5- Methyl-1,2,4-triazol-3-yl)-5,6-dihydroimidazo[1,2-d][1,4]benzoxazepine-9-yl]pyrazole- 1-yl]-2-methylacrylamide), MLN-1117 ((S)-methylphosphonic acid hydrogen (2R)-1-phenoxy-2-butyl ester; or methyl (side oxy ){[(2R)-l-phenoxy-2-butanyl]oxy}phosphonium)), BYL-719 ((2S)-N1-[4-methyl-5-[2-(2, 2,2-trifluoro-1,1-dimethylethyl)-4-pyridyl]-2-thiazolyl]-1,2-pyrrolidine dimethylamide), GSK2126458 (2,4-difluoro -N-{2-(methyloxy)-5-[4-(4-pyrazinyl)-6-quinolinyl]-3-pyridyl}benzenesulfonamide), TGX-221 ((± )-7-methyl-2-(morpholin-4-yl)-9-(l-phenylaminoethyl)-pyrido[1,2-a]-pyrimidin-4-one), GSK2636771 ( 2-methyl-1-(2-methyl-3-(trifluoromethyl)benzyl)-6-morpholinyl-1H-benzo[d]imidazole-4-carboxylic acid dihydrochloride), KIN -193 ((R)-2-((l-(7-methyl-2-morpholinyl-4-oxo-4H-pyrido[1,2-a]pyrimidin-9-yl)ethyl )amino)benzoic acid), TGR-1202/RP5264, GS-9820 ((S)-l-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4- Monohydroxypropan-1-one), GS-1101 (5-fluoro-3-phenyl-2-([S)]-1-[9H-purin-6-ylamino]-propyl)-3H- Quinazolin-4-one), AMG-319, GSK-2269557, SAR245409 (N-(4-(N-(3-((3,5-dimethoxyphenyl)amino)quinoxaline- 2-yl)sulfamoyl)phenyl)-3-methoxy-4-methylbenzamide), BAY80-6946 (2-amino-N-(7-methoxy-8-(3 -morpholinopropoxy)-2,3-dihydroimidazo[l,2-c]quinazoline), AS 252424 (5-[l-[5-(4-fluoro-2-hydroxy-benzene base)-furan-2-yl]-methyl-(Z)-ylidene]-thiazolidine-2,4-dione), CZ 24832 (5-(2-amino-8-fluoro-[1, 2,4]triazolo[l,5-a]pyridin-6-yl)-N-tert-butylpyridine-3-sulfonamide), buparlisib (5-[2,6 -bis(4-morpholinyl)-4-pyrimidinyl]-4-(trifluoromethyl)-2-pyridinylamine), GDC-0941 (2-(1H-indazol-4-yl)-6- [[4-(Methylsulfonyl)-1-hexahydropyrazinyl]methyl]-4-(4-morpholinyl)thieno[3,2-d]pyrimidine), GDC-0980 (( S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl ) Hexahydropyrazin-1-yl)-2-hydroxypropan-1-one (also known as RG7422)), SF1126 ((8S,14S,17S)-14-(carboxymethyl)-8-(3- Guanidinopropyl)-17-(hydroxymethyl)-3,6,9,12,15-pentaoxy-1-(4-(4-oxo-8-phenyl-4H-alkene -2-yl)morpholinyl-4-ium)-2-oxa-7,10,13,16-tetraazaoctadecane-18-ate), PF-05212384 (N-[4-[ [4-(Dimethylamino)-1-hexahydropyridyl]carbonyl]phenyl]-N'-[4-(4,6-di-4-morpholinyl-1,3,5-tri oxazin-2-yl)phenyl]urea), LY3023414, BEZ235 (2-methyl-2-{4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2 ,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl]phenyl}propionitrile), XL-765 (N-(3-(N-(3-(3,5 -dimethoxyphenylamino)quinoxalin-2-yl)sulfamoyl)phenyl)-3-methoxy-4-methylbenzamide) and GSK1059615 (5-[[4 -(4-pyridyl)-6-quinolinyl]methylene]-2,4-thiazolidinedione), PX886 (acetic acid [(3aR,6E,9S,9aR,10R,11aS)-6-[ [Bis(prop-2-enyl)amino]methylene]-5-hydroxy-9-(methoxymethyl)-9a,11a-dimethyl-1,4,7-trioxyl -2,3,3a,9,10,11-hexahydroindeno[4,5h]iso-alken-10-yl]ester (also known as sonolisib)) and described in WO2014/071109 Has a formula structure.

BTK inhibitors are well known. Examples of BTK inhibitors include ibrutinib (also known as PCI-32765) (Imbruvica™) (1-[(3R)-3-[4-amino-3-(4-phenoxy-phenyl) pyrazolo[3,4-d]pyrimidin-1-yl]hexahydropyridin-1-yl]prop-2-en-1-one), diphenylaminopyrimidine-based inhibitors (such as AVL-101 and AVL -291/292 (N-(3-((5-fluoro-2-((4-(2-methoxyethoxy)phenyl)amino)pyrimidin-4-yl)amino)phenyl) Acrylamide) (Avila Therapeutics)) (see U.S. Patent Publication No. 2011/0117073, which is incorporated herein in its entirety), Dasatinib ([N-(2-chloro-6-methylphenyl) -2-(6-(4-(2-Hydroxyethyl)hexahydropyrazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide], LFM-A13 (α-cyano-β-hydroxy-β-methyl-N-(2,5-dibromophenyl)acrylamide), GDC-0834 ([R-N-(3-(6-(4-(1 ,4-Dimethyl-3-oxohexahydropyrazin-2-yl)phenylamino)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl )-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide], CGI-560 4-(tert-butyl)-N-(3 -(8-(Phenylamino)imidazo[1,2-a]pyrazin-6-yl)phenyl)benzamide, CGI-1746 (4-(tert-butyl)-N-( 2-methyl-3-(4-methyl-6-((4-(morpholine-4-carbonyl)phenyl)amino)-5-oxo-4,5-dihydropyrazine-2 -yl)phenyl)benzamide), CNX-774 (4-(4-((4-((3-acrylamidophenyl)amino)-5-fluoropyrimidin-2-yl)amine base) phenoxy)-N-picoline carboxamide), CTA056 (7-benzyl-1-(3-(hexahydropyridin-1-yl)propyl)-2-(4-(pyridine- 4-yl)phenyl)-1H-imidazo[4,5-g]quinoxalin-6(5H)-one), GDC-0834 ((R)-N-(3-(6-((4 -(1,4-Dimethyl-3-oxahydropyrazin-2-yl)phenyl)amino)-4-methyl-5-oxo-4,5-dihydropyrazine -2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide), GDC-0837 ((R)-N-(3 -(6-((4-(1,4-Dimethyl-3-oxahydropyrazin-2-yl)phenyl)amino)-4-methyl-5-oxo-4 ,5-dihydropyrazin-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide), HM-71224, ACP -196, ONO-4059 (Ono Pharmaceuticals), PRT062607 (4-((3-(2H-1,2,3-triazol-2-yl)phenyl)amino)-2-(((1R,2S )-2-aminocyclohexyl)amino)pyrimidine-5-formamide hydrochloride), QL-47 (1-(1-acryloylindolin-6-yl)-9-(1 -methyl-1H-pyrazol-4-yl)benzo[h][1,6]naphthyridin-2(1H)-one) and RN486 (6-cyclopropyl-8-fluoro-2-(2 -Hydroxymethyl-3-{1-methyl-5-[5-(4-methyl-hexahydropyrazin-1-yl)-pyridin-2-ylamino]-6-oxo-1 , 6-dihydro-pyridin-3-yl}-phenyl)-2H-isoquinolin-1-one) and other molecules capable of inhibiting BTK activity (such as they are disclosed in Akinleye et al., Journal of Hematology & Oncology , 2013, BTK inhibitors in 6:59, the entire content of which is incorporated herein by reference).

Syk inhibitors are well known and include, for example, Cerdulatinib (4-(cyclopropylamino)-2-((4-(4-(ethylsulfonyl)hexahydropyrazine- 1-yl) phenyl) amino) pyrimidine-5-formamide), entotinib (entospletinib) (6-(1H-indazol-6-yl)-N-(4-morpholinylphenyl ) imidazo[1,2-a]pyrazin-8-amine), fostamatinib (dihydrogen phosphate [6-({5-fluoro-2-[(3,4,5-trimethoxy phenyl)amino]-4-pyrimidinyl}amino)-2,2-dimethyl-3-oxo-2,3-dihydro-4H-pyrido[3,2-b][ 1,4]oxazin-4-yl]methyl ester), fotatinib disodium salt ((6-((5-fluoro-2-((3,4,5-trimethoxyphenyl)amine Base) pyrimidin-4-yl) amino) -2,2-dimethyl-3-oxo-2H-pyrido[3,2-b][1,4]oxazine-4(3H)- base) sodium methyl phosphate), BAY 61-3606 (2-(7-(3,4-dimethoxyphenyl)-imidazo[1,2-c]pyrimidin-5-ylamino)-nicotin Alkali amide hydrochloride), RO9021 (6-[(1R,2S)-2-amino-cyclohexylamino]-4-(5,6-dimethyl-pyridin-2-ylamino)- pyrazine-3-carboxyamide), imatinib (Gleevec; 4-[(4-methylhexahydropyrazin-1-yl)methyl]-N-(4-methyl-3-{[4 -(pyridin-3-yl)pyrimidin-2-yl]amino}phenyl)benzamide), staurosporine (staurosporine), GSK143 (2-(((3R,4R)-3-amine Basetetrahydro-2H-pyran-4-yl)amino)-4-(p-tolylamino)pyrimidine-5-carboxamide), PP2 (1-(tert-butyl)-3-( 4-chlorophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine), PRT-060318 (2-(((1R,2S)-2-aminocyclohexyl)amino) -4-(m-tolylamino)pyrimidine-5-formamide), PRT-062607 (4-((3-(2H-1,2,3-triazol-2-yl)phenyl)amine base)-2-(((1R,2S)-2-aminocyclohexyl)amino)pyrimidine-5-carboxamide hydrochloride), R112 (3,3'-((5-fluoropyrimidine-2 ,4-diyl)bis(azanediyl))diphenol), R348 (3-ethyl-4-methylpyridine), R406 (6-((5-fluoro-2-((3,4, 5-trimethoxyphenyl)amino)pyrimidin-4-yl)amino)-2,2-dimethyl-2H-pyrido[3,2-b][1,4]oxazine-3( 4H)-keto), YM193306 (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643), 7-azaindole, Pitrosine Alcohol (piceatannol), ER-27319 (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, which is incorporated herein in its entirety), compound D (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, the entire contents of which are incorporated herein), PRT060318 (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, the entire contents of which are incorporated herein), luteolin (luteolin) (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J.Med.Chem. 2012, 55, 3614-3643, its entirety is incorporated herein), apigenin (apigenin) (referring to the people such as Singh, Discovery and Development of Spleen Tyrosine Kinase ( SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, the entire contents of which are incorporated herein), quercetin (quercetin) (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors , J. Med. Chem. 2012, 55, 3614-3643, the entire contents of which are incorporated herein), fisetin (fisetin) (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, the entire contents of which are incorporated herein), myricetin (myricetin) (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem . 2012, 55, 3614-3643, the entire contents of which are incorporated herein), morin (morin) (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55 , 3614-3643, incorporated herein in its entirety).

The chemotherapeutic agent can also be a B-cell lymphoma 2 (Bcl-2) protein inhibitor. BCL-2 inhibitors are known in the art and include, for example, ABT-199 (4-[4-[[2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-ene -1-yl]methyl]hexahydropyrazin-1-yl]-N-[[3-nitro-4-[[(tetrahydro-2H-pyran-4-yl)methyl]amino] Phenyl]sulfonyl]-2-[(1H-pyrrolo[2,3-b]pyridin-5-yl)oxy]benzamide), ABT-737 (4-[4-[[2 -(4-chlorophenyl)phenyl]methyl]hexahydropyrazin-1-yl]-N-[4-[[(2R)-4-(dimethylamino)-1-phenylthio Alkylbutan-2-yl]amino]-3-nitrophenyl]sulfonylbenzamide), ABT-263 ((R)-4-(4-((4'-chloro-4 ,4-Dimethyl-3,4,5,6-tetrahydro-[l, l'-biphenyl]-2-yl)methyl)hexahydropyrazin-1-yl)-N-((4 -((4-morpholinyl-1-(phenylthio)butan-2-yl)amino)-3((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzoyl amine), GX15-070 (obatoclax mesylate, (2Z)-2-[(5Z)-5-[(3,5-dimethyl-lH-pyrrol-2-yl) Methyl]-4-methoxypyrrole-2-ylidene]indole-methanesulfonic acid))), 2-methoxy-antimycin A3, YC137 (4-(4,9-dioxo -4,9-dihydronaphtho[2,3-d]thiazol-2-ylamino)-phenyl ester), gossypol (pogosin), 2-amino-6-bromo-4-(1- Cyano-2-ethoxy-2-oxoethyl)-4H-methene-3-ethyl carboxylate, nilotinib-d3, TW-37 (N-[4-[[2-( 1,1-Dimethylethyl)phenyl]sulfonyl]phenyl]-2,3,4-trihydroxy-5-[[2-(1-methylethyl)phenyl]methyl] benzamide), Apogossypolone (ApoG2), or G3139 (Oblimersen).

Other chemotherapeutic agents for use in the methods contemplated herein include, but are not limited to, midazolam, MEK inhibitors, RAS inhibitors, ERK inhibitors, ALK inhibitors, HSP inhibitors (such as HSP70 and HSP 90 inhibitors or combinations thereof), RAF inhibitors, apoptosis compounds, topoisomerase inhibitors, AKT inhibitors (including but not limited to) MK-2206, GSK690693, perifosine (KRX-0401), GDC-0068, Triciribine, AZD5363, Honokiol, PF-04691502, and Miltefosine) or FLT-3 inhibitors (including but not limited to) P406, multiple Virtinib, Quizartinib (AC220), Amuvatinib (MP-470), Tandutinib (MLN518, ENMD-2076 and KW-2449) or combinations thereof . Examples of MEK inhibitors include, but are not limited to, trametinib/GSK1120212 (N-(3-{3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino] -6,8-Dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidine-l(2H-yl}phenyl) Acetamide), selumetinib (6-(4-bromo-2-chloroanilino)-7-fluoro-N-(2-hydroxyethoxy)-3-methylbenzimidazole- 5-formamide), pimasertib/AS703026/MSC1935369 ((S)-N-(2,3-dihydroxypropyl)-3-((2-fluoro-4-iodophenyl) Amino)isonicotinamide), XL-518/GDC-0973 (l-({3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl}carbonyl )-3-[(2S)-hexahydropyridin-2-yl]azetidin-3-ol), Rifatinib (refametinib)/BAY869766/RDEA119 (N-(3,4-difluoro-2 -(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide), PD-0325901 ( N-[(2R)-2,3-dihydroxypropoxy]-3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-benzamide), TAK733 ((R)-3-(2,3-dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3d]pyrimidine -4,7(3H,8H)-dione), MEK162/ARRY438162 (5-[(4-bromo-2-fluorophenyl)amino]-4-fluoro-N-(2-hydroxyethoxy) -1-methyl-1H-benzimidazole-6 formamide), R05126766 (3-[[3-fluoro-2-(methylsulfamoylamino)-4-pyridyl]methyl]- 4-methyl-7-pyrimidin-2-yloxy-alken-2-one), WX-554, R04987655/CH4987655 (3,4-difluoro-2-((2-fluoro-4-iodophenyl )amino)-N-(2-hydroxyethoxy)-5-((3-side oxy-l,2-oxazinyl-2 base)methyl)benzamide) or AZD8330 (2- ((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxo-1,6-dihydropyridine-3 - formamide). Examples of RAS inhibitors include, but are not limited to, Reolysin and siG12D LODER. Examples of ALK inhibitors, include, but are not limited to, crizotinib, AP26113, and LDK378. HSP Inhibitors include, but are not limited to, Geldanamycin or 17-N-allylamino-17-desmethoxygeldanamycin (17AAG) and Radicicol .

Known ERK inhibitors include SCH772984 (Merck/Schering-Plough), VTX-11e (Vertex), DEL-22379, Ulixertinib (BVD-523, VRT752271), GDC-0994, FR 180204, XMD8-92 and ERK5-IN-1.

Raf inhibitors are well known and include, for example, Vemurafinib (N-[3-[[5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3 -yl]carbonyl]-2,4-difluorophenyl]-1-propanesulfonamide), sorafenib tosylate (4-[4-[[4-chloro-3-(trifluoromethyl )phenyl]carbamoylamino]phenoxy]-N-methylpyridine-2-formamide; 4-methylbenzenesulfonate), AZ628 (3-(2-cyanopropane-2 -yl)-N-(4-methyl-3-(3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylamino)phenyl)benzamide) , NVP-BHG712 (4-methyl-3-(1-methyl-6-(pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino)-N -(3-(trifluoromethyl)phenyl)benzamide), RAF-265 (1-methyl-5-[2-[5-(trifluoromethyl)-1H-imidazol-2-yl ]pyridin-4-yl]oxy-N-[4-(trifluoromethyl)phenyl]benzimidazol-2-amine), 2-bromoaldisine (2-Bromoaldisine) (2-bromo-6 ,7-dihydro-1H,5H-pyrrolo[2,3-c]azepine-4,8-dione), Raf kinase inhibitor IV (2-chloro-5-(2-phenyl-5- (pyridin-4-yl)-1H-imidazol-4-yl)phenol) and sorafenib N- oxide (4-[4-[[[[4-chloro-3(trifluoromethyl)phenyl ]amino]carbonyl]amino]phenoxy]-N-methyl-2-pyridinecarboxamide 1-oxide).

Known topoisomerase I inhibitors useful in the present invention include (S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyran And[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H,12H)-dione monohydrochloride (topotecan), (S) -4-Ethyl-4-hydroxy-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14-(4H,12H)-di Ketone (camptothecin), (1S,9S)-1-amino-9-ethyl-5-fluoro-1,2,3,9,12,15-hexahydro-9-hydroxy-4-methyl -10H,13H-benzo(de)pyrano(3',4':6,7)indazino(1,2-b)quinoline-10,13-dione (exatecan )), (7-(4-methylhexahydropyrazinylmethylene)-10,11-ethylenedioxy-20(S)-camptothecin (lurotecan) or (S)-4,11-diethyl-3,4,12,14-tetrahydro-4-hydroxy-3,14-dioxo 1H-pyrano[3',4':6,7 ]-indazino[1,2-b]quinolin-9-yl-[1,4'bishexahydropyridine]-1'-carboxylate (irinotecan), (R)-5-ethyl -9,10-difluoro-5-hydroxy-4,5-dihydroxyl[3',4':6,7]indazino[1,2-b]quinoline-3,15(1H ,13H)-diketone (diflumotecan), (4S)-11-((E)-((1,1-dimethylethoxy)imino)methyl)-4-ethyl -4-Hydroxy-1,12-dihydro-14H-pyrano(3',4':6,7)indazino(1,2-b)quinoline-3,14(4H)-dione (gimatecan (gimatecan)), (S)-8-ethyl-8-hydroxyl-15-((4-methylhexahydropyrazin-1-yl)methyl)-11,14-dihydro- 2H-[1,4]dioxino[2,3-g]pyrano[3',4':6,7]indazino[1,2-b]quinoline-9, 12(3H,8H)-Diketone (Letotecan), (4S)-4-Ethyl-4-Hydroxy-11-[2-[(1-methylethyl)amino]ethyl]- 1H-Pyrano[3?,4?:6,7]indazino[1,2-b]quinoline-3,14(4H,12H)-dione (belotecan), 6-((1,3-dihydroxypropan-2-yl)amino)-2,10-dihydroxy-12-((2R,3R,4S,5S,6R)-3,4,5-trihydroxy -6-(Hydroxymethyl)tetrahydro-2H-pyran-2-yl)-12,13-dihydro-5H-indolo[2,3-a]pyrrolo[3-c]carbazole- 5,7(6H)-diketone (edotecarin), 8,9-dimethoxy-5-(2-N,N-dimethylaminoethyl)-2,3- Methylenedioxy-5H-dibenzo(c,h)(1,6)naphthyridin-6-one (topovale), benzo[6,7]indazino[1, 2-b] Quinolin-11(13H)-one (rosettacin), (S)-4-ethyl-4-hydroxy-11-(2-(trimethylsilyl)ethyl) -1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H,12H)-dione (cositecan) , N, N', N'', N'''-{methanetetrayltetra[methylene poly(oxyethylidene)oxy(1-pentoxyethylidene)]}tetraglycine tetra{ (4S)-9-[([1,4'-bis-hexahydropyridyl]-1'-carbonyl)oxy]-4,11-diethyl-3,14-dioxo-3,4 , 12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-4-yl}ester tetrahydrochloride (polyethylene glycol Alcoholated irinotecan (etirinotecan pegol)), 10-hydroxy-camptothecin (HOCPT), 9-nitrocamptothecin (rubitecan), SN38 (7-ethyl-10-hydroxycamptothecin base) and 10-hydroxy-9-nitrocamptothecin (CPT109), (R)-9-chloro-5-ethyl-5-hydroxy-10-methyl-12-((4-methylhexahydro Pyridin-1-yl)methyl)-4,5-dihydroxyl[3',4':6,7]indazino[1,2-b]quinoline-3,15(1H,13H ) - diketone (elmotecan).

Treatment regimens In some aspects, treatment regimens are administered to patients with advanced or metastatic cancer whose disease has progressed (indicative of immune checkpoint inhibitor resistance) following prior treatment with a PD-1 or PD-L1 inhibitor. Therefore, these patients are administered treacilil, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors, wherein other checkpoint inhibitors are selected from TIGIT inhibitors, TIM-3 inhibitors, LAG- 3 inhibitors or CD73 inhibitors. In some embodiments, straciclib, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors are administered on Day 1 of each 14-day therapeutic treatment cycle (or cycles), and at Trascilli was administered again on Day 7 of each 14-day cycle. In some embodiments, treracilib is administered one or more times per week. In some embodiments, straciclib, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors are administered on Day 1 of each 21-day cycle, and administered on Days 7 and 10 of each 21-day cycle. On the 14th day, Traxil was administered again. In some embodiments, treracilib is administered one or more times per week. In some embodiments, straciclib, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors are administered on Day 1 of each 28-day cycle, and administered on Days 7, 10 of each 28-day cycle. On the 14th day and the 21st day, Trascilli was administered again. In some embodiments, treracilib is administered one or more times per week. In some embodiments, treracilil, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors are administered on Day 1 of each 42-day cycle, and on Day 7, On the 14th day, the 21st day, the 28th day and the 35th day, treracilib was administered again. In some embodiments, treracilib is administered one or more times per week. In some embodiments, treracillib is administered once a week during the treatment period, and is administered once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, or once every six weeks PD-1 or PD-L1 and other immune checkpoint inhibitors. In some embodiments, triaciclib is administered weekly, PD-L1 or PD-1 is administered according to its standard administration label for its approved use, and the other immune checkpoint inhibitor is administered in combination with PD- L1 or PD-1 inhibitors are administered concurrently. In some embodiments, triracillib is administered once a week, and once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, or once every twelve weeks Co-formulations of immune checkpoint inhibitors are administered in an effective amount. In some embodiments, co-formulations of immune checkpoint inhibitors include PD-L1 or PD-1 immune checkpoint inhibitors and other immune checkpoint inhibitors selected from the group consisting of TIGIT inhibitors, TIM-3 Inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, PD-L1 or PD-1 immune checkpoint inhibitors and other immune checkpoint inhibitors are administered in varying amounts. In some embodiments, the PD-L1 or PD-1 immune checkpoint inhibitor and the other immune checkpoint inhibitor are administered in the same amount. In some embodiments, the other immune checkpoint inhibitor is a TIGIT inhibitor. In some embodiments, the other immune checkpoint inhibitor is a TIM-3 inhibitor. In some embodiments, the other immune checkpoint inhibitor is a LAG-3 inhibitor. In some embodiments, treracillib is administered weekly and an effective amount of opdualag is administered once every four weeks. In some embodiments, the opdualag is a co-formulation comprising about 480 mg of nivolumab and about 160 mg of relarimab. In some embodiments, opdualag is administered as a single 30-minute infusion. In some embodiments, opdualag is administered every 4 weeks. In some embodiments, the other immune checkpoint inhibitor is a CD73 inhibitor. In some embodiments, other immune checkpoint inhibitors are not administered concomitantly with the PD-L1 or PD-1 inhibitor. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The LAG-3 immune checkpoint inhibitor is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, or every twelve weeks. In some embodiments, the LAG-3 immune checkpoint inhibitor is Rilarimab. In some embodiments, treracicill is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and rilarimab Administer in accordance with its standard administration labeling for its approved use.

In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The TIGIT immune checkpoint inhibitor is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, or every twelve weeks. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The TIM-3 immune checkpoint inhibitor is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, or every twelve weeks. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The CD73 immune checkpoint inhibitor is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, or every twelve weeks. In some embodiments, only the initial loading dose of treracilil is administered about 8 days, 7 days, 6 days, 5 days, 4 days, or 3 days prior to beginning the first treatment cycle as set forth above. In some embodiments, the cancer is non-small cell lung cancer (NSCLC), triple negative breast cancer (TNBC), metastatic colorectal cancer (CRC), or metastatic urothelial carcinoma (mUC). In some embodiments, the patient has second-line metastatic non-squamous or squamous NSCLC. In some embodiments, the patient has second-line metastatic triple-negative breast cancer. In some embodiments, the patient has second line metastatic colorectal cancer (CRC). In some embodiments, the patient has second-line locally advanced or metastatic urothelial carcinoma (mUC). In some alternative embodiments, the other immune checkpoint inhibitor administered in combination with treracicill and a PD-1 or PD-L1 inhibitor is selected from the group consisting of programmed death ligand 2 (PD-L2), CTLA-4, and V domain Ig inhibitor of T cell activation (VISTA), B7-H3/CD276, indoleamine 2,3-dioxygenase (IDO), killer immunoglobulin-like receptor (KIR), carcinoembryonic antigen cells Inhibitors of adhesion molecules (CEACAM) (such as CEACAM-1, CEACAM-3, and CEACAM-5), sialic acid-binding immunoglobulin-like lectin 15 (Siglec-15), and B and T lymphocyte attenuation protein (BTLA). In an alternative to the above embodiment, the treatment regimen administers treracilil in combination with PD-1 or PD-L1 as described herein and various tyrosine kinase (MTK) inhibitors (in place of other immune checkpoint inhibitors) , such as (but not limited to) Levatinib, Stratinib and Cabozantinib). In some embodiments, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15 , at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24 or more treatment cycles. In some embodiments, the patient is administered treatment cycles until disease progression. In some embodiments, treracilib is administered one or more times per week. In some embodiments, treracillib is administered once a week during the treatment period, and is administered once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, or once every six weeks A PD-1 or PD-L1 immune checkpoint inhibitor and other immune checkpoint inhibitors administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, or every six weeks . In some embodiments, triraciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint inhibitor is administered The agent is administered concurrently with PD-L1 or PD-1 inhibitors. In some embodiments, other immune checkpoint inhibitors are not administered concomitantly with the PD-L1 or PD-1 inhibitor. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered Administer in accordance with its standard administration labeling for its approved use. In some embodiments, only the initial loading dose of treracilil is administered about 8 days, 7 days, 6 days, 5 days, 4 days, or 3 days prior to beginning the first treatment cycle as set forth above. In some embodiments, the cancer is non-small cell lung cancer (NSCLC), triple negative breast cancer (TNBC), metastatic colorectal cancer (CRC), or metastatic urothelial carcinoma (mUC). In some embodiments, the patient has second-line metastatic non-squamous or squamous NSCLC. In some embodiments, the patient has second-line metastatic triple-negative breast cancer. In some embodiments, the patient has second line metastatic colorectal cancer (CRC). In some embodiments, the patient has second-line locally advanced or metastatic urothelial carcinoma (mUC). In some alternative embodiments, the other immune checkpoint inhibitor administered in combination with treracicill and a PD-1 or PD-L1 inhibitor is selected from the group consisting of programmed death ligand 2 (PD-L2), CTLA-4, and V domain Ig inhibitor of T cell activation (VISTA), B7-H3/CD276, indoleamine 2,3-dioxygenase (IDO), killer immunoglobulin-like receptor (KIR), carcinoembryonic antigen cells Inhibitors of adhesion molecules (CEACAM) (such as CEACAM-1, CEACAM-3, and CEACAM-5), sialic acid-binding immunoglobulin-like lectin 15 (Siglec-15), and B and T lymphocyte attenuation protein (BTLA). In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The PD-L2 immune checkpoint inhibitor is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, or every six weeks. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The CTLA-4 immune checkpoint inhibitor is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, or every twelve weeks. In some embodiments, the CTLA-4 immune checkpoint inhibitor is ipilimumab. In some embodiments, triracillib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and ipilimumab is administered Administer in accordance with its standard administration labeling for its approved use. In some embodiments, triracillib is administered weekly, and the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and once every three weeks Administer ipilimumab. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The VISTA immune checkpoint inhibitor is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, or every twelve weeks. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The B7-H3/CD276 immune checkpoint inhibitor is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, or every twelve weeks. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The IDO immune checkpoint inhibitor is administered once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, or once every twelve weeks. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The KIR immune checkpoint inhibitor is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, or every twelve weeks. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The CEACAM immune checkpoint inhibitor is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, or every twelve weeks. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The Siglec-15 immune checkpoint inhibitor is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, or every twelve weeks. In some embodiments, triaciclib is administered weekly, the PD-L1 or PD-1 immune checkpoint inhibitor is administered according to its standard administration label for its approved use, and the other immune checkpoint is administered The BTLA immune checkpoint inhibitor is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, or every twelve weeks. In an alternative embodiment to the above embodiment, the treatment regimen administers treracilil and a PD-1 or PD-L1 immune checkpoint inhibitor as described herein and various tyrosine kinase (MTK) inhibitors (in place of other Immune checkpoint inhibitors such as (but not limited to) levatinib, stretratinib and cabozantinib). In some embodiments, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15 , at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24 or more treatment cycles. In some embodiments, patients are administered treatment cycles at the doses set forth herein until disease progression. As contemplated herein, specifically timed administration of triracillib in combination with one or more chemotherapeutic agents, PD-1 or PD-L1 immune checkpoint inhibitors and other immune checkpoint inhibitors (wherein the other checkpoint inhibitors are selected TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors), and thus G0/G1 arrest induced by treracicill are short-term and transient. Cells quiescent in the G1 phase of the cell cycle are more resistant to the damaging effects of chemotherapeutic agents than proliferating cells. Treatment regimens are specific to the solid tumor being treated. In some embodiments, less than 24 hours, less than 16 hours, less than 12 hours, less than 8 hours, less than 6 hours, less than 4 hours prior to administration of one or more chemotherapeutic agents , less than 2 hours, less than 1 hour, or about 30 minutes) and administer a PD-1 or PD-L1 inhibitor and other immune checkpoint inhibitor on Day 1 of each therapeutic treatment cycle, Wherein other checkpoint inhibitors are selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors.

In some aspects, the treatment regimen is administered to patients with advanced or metastatic cancer whose disease has progressed (indicative of immune checkpoint inhibitor resistance) following prior treatment with a PD-1 or PD-L1 inhibitor. Therefore, these patients are administered treacilil, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors, wherein other checkpoint inhibitors are selected from TIGIT inhibitors, TIM-3 inhibitors, LAG- 3 inhibitors or CD73 inhibitors. In some embodiments, straciclib, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors are administered on Day 1 of each 14-day therapeutic treatment cycle (or cycles), and at Trascilli was administered again on Day 7 of each 14-day cycle. In some embodiments, treracilib is administered one or more times per week. In some embodiments, straciclib, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors are administered on Day 1 of each 21-day cycle, and administered on Days 7 and 10 of each 21-day cycle. On the 14th day, Traxil was administered again. In some embodiments, treracilib is administered one or more times per week. In some embodiments, straciclib, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors are administered on Day 1 of each 28-day cycle, and administered on Days 7, 10 of each 28-day cycle. On the 14th day and the 21st day, Trirascillin was administered again. In some embodiments, treracilib is administered one or more times per week. In some embodiments, treracilil, a PD-1 or PD-L1 inhibitor, and other immune checkpoint inhibitors are administered on Day 1 of each 42-day cycle, and on Day 7, On the 14th day, the 21st day, the 28th day and the 35th day, treracilib was administered again. In some embodiments, treracilib is administered one or more times per week. In some embodiments, treracillib is administered once a week during the treatment period, and is administered once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, or once every six weeks PD-1 or PD-L1 and other immune checkpoint inhibitors. In some embodiments, triaciclib is administered weekly, PD-L1 or PD-1 is administered according to its standard administration label for its approved use, and the other immune checkpoint inhibitor is administered in combination with PD- L1 or PD-1 inhibitors are administered concurrently. In some embodiments, the other immune checkpoint inhibitor is not administered concomitantly with the PD-L1 or PD-1 inhibitor. In some embodiments, only the initial loading dose of treracilil is administered about 8 days, 7 days, 6 days, 5 days, 4 days, or 3 days prior to beginning the first treatment cycle as set forth above. In some embodiments, the cancer is non-small cell lung cancer (NSCLC), triple negative breast cancer (TNBC), colorectal cancer (CRC), urothelial carcinoma (mUC), or another solid tumor. In some embodiments, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15 , at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24 or more treatment cycles. In some embodiments, the patient is administered treatment cycles until disease progression.

In some alternative embodiments, the other immune checkpoint inhibitor administered in combination with treracicill and a PD-1 or PD-L1 inhibitor is selected from the group consisting of programmed death ligand 2 (PD-L2), CTLA-4, and V domain Ig inhibitor of T cell activation (VISTA), B7-H3/CD276, indoleamine 2,3-dioxygenase (IDO), killer immunoglobulin-like receptor (KIR), carcinoembryonic antigen cells Inhibitors of adhesion molecules (CEACAM) (such as CEACAM-1, CEACAM-3, and CEACAM-5), sialic acid-binding immunoglobulin-like lectin 15 (Siglec-15), and B and T lymphocyte attenuation protein (BTLA). In an alternative to the above embodiment, the treatment regimen administers treracilil in combination with PD-1 or PD-L1 as described herein and various tyrosine kinase (MTK) inhibitors (in place of other immune checkpoint inhibitors) , such as (but not limited to) Levatinib, Stratinib and Cabozantinib).

Non-Small Cell Lung Cancer Non-small cell lung cancer makes up approximately 85% of all lung cancer diagnoses. Common types of non-small cell lung cancer include adenocarcinomas (which typically exhibit glandular differentiation), squamous cell carcinomas (which typically exhibit squamous differentiation (keratinization)), and large cell carcinomas (which typically appear large and poorly differentiated). Such patients should undergo molecular testing for oncogenes and programmed death-ligand 1 (PD-L1). Patients with advanced or recurrent disease with an active oncogene should be considered for treatment with targeted therapy. Patients without an actionable genetic alteration should be treated with treracilib and chemotherapy alone, treracilib, chemotherapy and immunotherapy, or treracilib and immunotherapy only. Patients with poor performance status or comorbidities may consider single-agent chemotherapy, immunotherapy, or (if they have an active oncogene) targeted therapy.

Squamous NSCLC In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic squamous NSCLC in a first-line advanced/metastatic setting, wherein the patient is administered Qu Rascilli, chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic squamous cell NSCLC in the first-line advanced/metastatic setting, wherein the patient is administered triaciclib, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic squamous cell NSCLC in a second-line advanced/metastatic setting, wherein the patient is administered triaciclib , chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic squamous cell NSCLC in a second-line advanced/metastatic setting, wherein the patient is administered triaciclib , PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic squamous cell NSCLC in a second-line advanced/metastatic setting, wherein the patient is administered triaciclib , PD-1 or PD-L1 inhibitors and multiple tyrosine kinase (MTK) inhibitors. In some embodiments, the MTK inhibitor is selected from levatinib, stretratinib, or crizotinib. In some embodiments, the patient has a tumor expressing PD-L1. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test.

In one embodiment, CDK4/6 inhibitors and PD-1 or PD-L1 checkpoint inhibitors and other immune checkpoint inhibitors (ICI) (selected from T cell immune receptors with Ig and ITIM domains (TIGIT ) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint inhibitor, or cluster of differentiation 73 (CD73) checkpoint inhibitor ) can be used in combination with many standard-of-care chemotherapy regimens such as, but not limited to, first-line or second-line treatment regimens for squamous cell NSCLC, such as, but not limited to, single-agent chemotherapy or combination chemotherapy. Non-limiting examples of single-agent chemotherapy regimens for first-line treatment of locally advanced or metastatic squamous cell NSCLC include 50 mg/ ^m IV cisplatin on days 1, 8, 29, and 36 + days 1-5 and 50 mg/m ² IV etoposide on days 29-33; 100 mg/m ² IV cisplatin + 5 mg/m ² /week IV vinblastine on days 1 and 29 for 5 weeks; weekly AUC 2 IV carboplatin and For 7 weeks + 45-50 mg/ ^m2 IV paclitaxel weekly for 7 weeks, followed by two cycles with Day 1 AUC 6 IV carboplatin + 200 mg/ ^m2 IV paclitaxel on Day 1 every 21 weeks Consolidation chemotherapy; 200 mg/ ^m2 IV paclitaxel every 21 days, 35 mg/ ^m2 IV docetaxel every 28 days for 3 weeks, 75 mg/ ^m2 IV docetaxel every 21 days , 1000 mg/m ² IV gemcitabine on days 1, 8 and 15 every 4 weeks, 1250 mg/m 2 IV gemcitabine on days 1 and 8 every 21 days, 25 mg/ ^{m 2 IV} vinorelbine every week, every 21 days 30 mg/m ² IV vinorelbine on days 1 and 8. A non-limiting example of a combination chemotherapy regimen for first-line treatment of locally advanced or metastatic squamous cell NSCLC includes 75 mg/m IV ^cisplatin on day 1 + 175 mg/ ^m IV pacific on day 1 every 21 days Paclitaxel, 100 mg/m ² IV cisplatin on day 1 + 1000 mg/m ² IV gemcitabine on days 1, 8, and 15 every 28 days, 60 mg/m ² IV cisplatin on day 1 + every 21 days on days 1 and 15 1000 mg/ ^m2 IV gemcitabine on day 8, 75 mg/ ^m2 IV cisplatin on day 1 + 75 mg/ ^m2 IV docetaxel on day 1 every 21 days, AUC on day 1 6 IV carboplatin + every 21 days Day 175-225 mg/m ² IV paclitaxel on day 1, AUC on day 1 6 IV carboplatin + 90 mg/m ² IV paclitaxel on days 1, 8 and 15 every 28 days, days 1 and 8 every 21 days And 100 mg/m ² IV protein-bound paclitaxel for 15 days + AUC 6 IV carboplatin on day 1, AUC 6 IV carboplatin on day 1 + 75 mg/m ² IV docetaxel on day 1 every 21 days, Day 1 AUC 5 IV carboplatin + 1250 mg/m ² IV gemcitabine on days 1 and 8 every 21 days, 100 mg ^{/m 2 IV cisplatin on day 1 every 28 days + 25 mg/m 2 IV} vinorel every week Vinorelbine, 40 mg/m ² IV cisplatin on day 1 + 25 mg/m ² IV vinorelbine on days 1 and 8 every 21 days or AUC 5 IV carboplatin on day 1 + 30 mg/m 2 IV carboplatin on days 1 and 8 every 21 days mg/m ² IV vinorelbine. A non-limiting example of a chemotherapy regimen for second-line treatment of locally advanced or metastatic squamous cell NSCLC includes 75 mg/m ² IV docetaxel on Day 1 every 21 days for 4 to 6 cycles +/ - 10 mg/kg IV ramucirumab.

In one embodiment, the CDK4/6 inhibitor is treracicil. In some embodiments, treracilib is administered less than 4 hours prior to administration of the first-line chemotherapy regimen. In some embodiments, treracicill is administered about one hour or less (eg, about 45 minutes, about 40 minutes, about 35 minutes, or about 30 minutes) prior to the administration of the first-line chemotherapy regimen. In some embodiments, the triracicil is administered intravenously to the patient at about 190 mg/m ² to 280 mg/m ² . In some embodiments, treracilil is administered at about 240 mg/m ² .

In one embodiment, the immune checkpoint inhibitor is a PD-1 inhibitor. In one embodiment, the immune checkpoint inhibitor is a PD-1 inhibitor selected from the group consisting of: nivolumab (OPDIVO ^® ), pembrolizumab (KEYTRUDA ^® ), cimipril anti (LIBTAYO ^® ) and dotalimab (JEMPERLI ^® ). In one embodiment, the PD-1 inhibitor is Nivolumab. In some embodiments, nivolumab is administered with platinum-containing chemotherapy at 360 mg on the same day every 3 weeks for 3 cycles. In some embodiments, nivolumab is administered at 240 mg every 2 weeks or at 480 mg every 4 weeks. In one embodiment, the PD-1 inhibitor is pembrolizumab. In some embodiments, pembrolizumab is administered at 200 mg every 3 weeks or at 400 mg every 6 weeks. In one embodiment, the PD-1 inhibitor is simiprizumab. In some embodiments, cimiprizumab 350 mg is administered as an intravenous infusion over 30 minutes every 3 weeks. In one embodiment, the PD-1 inhibitor is dotalimab. In some embodiments, dotalimab is administered as an intravenous infusion over 30 minutes at 500 mg every 3 weeks (for doses 1-4) and then at 1,000 mg every 6 weeks.

In one embodiment, the immune checkpoint inhibitor is a PD-L1 inhibitor. In one embodiment, the PD-L1 inhibitor is selected from the group consisting of atezolizumab (TECENTRIQ ^® ), avelumab (BAVENCIO ^® ) and durvalumab (IMFINZI ^® ). In one embodiment, the PD-LI inhibitor is atezolizumab. In some embodiments, atezolizumab is administered at 840 mg every 2 weeks, 1200 mg every 3 weeks, or 1680 mg every 4 weeks for up to 1 year. In some embodiments, the PD-L1 inhibitor is avelumab. In some embodiments, avelumab 800 mg is administered as an intravenous infusion over 60 minutes every 2 weeks. In one embodiment, the PD-L1 inhibitor is durvalumab. In some embodiments, administered at 10 mg/kg every 2 weeks or at 1500 mg every 4 weeks (for patients weighing greater than 30 kg) and at 10 mg/kg every 2 weeks (for patients weighing less than 30 kg) with durvalumab.

In one embodiment, the other checkpoint inhibitor is a TIGIT checkpoint inhibitor. In one embodiment, the TIGIT checkpoint inhibitor is selected from the group consisting of vebolizumab (MK-7684; Merck), etigimumab/OMP-313 M32 (OncoMed), tisreliumab Anti-(MTIG7192A/RG-6058; Roche/Genentech), Ospolizumab (BGB-A1217; Beigene), BMS-986207 (BMS), COM902 (Compugen), M6223 (Merck KGaA), Dovanarumab (AB-154; Arcus Biosciences), AZD2936 (AstraZeneca), JS006 (Shanghai Junshi Bioscience), IBI139 (Innovent Biologics), ASP-8374 (Astellas/Potenza), BAT6021 (Bio-Thera Solutions), TAB006 (Shanghai Junshi Bioscience) , Domvanalimab (AB154; Arcus Biosciences), EOS884448 (EOS-448; iTeos), SEA-TGT (Seattle Genetics), mAb-7 (Stanwei Biotech), SHR-1708 (Hengrui Medicine), GS02 (Suzhou Zelgen/Qilu Pharma) , RXI-804 (Rxi Pharmaceuticals), NB6253 (Northern Biologics), ENUM009 (Enumreal Biomedical), CASC-674 (Cascadian Therapeutics), AJUD008 (AJUD Biopharma), AGEN1777 (Agenus, Bristol-Myers Squibb), HLX53 (Squibb) hanghai Henlius Biotech ), BAT6005 (Bio-Thera Solutions), anti-TIGIT/anti-PD-L1 bispecific antibody HLX301 (Shanghai Henlius Biotech) and anti-TIGIT/anti-PD-L1 antibody HB0036 (Shanghai Huaota Biopharmaceutical).

In one embodiment, the other checkpoint inhibitor is a TIM-3 checkpoint inhibitor. In one embodiment, the TIM-3 checkpoint inhibitor is selected from the group consisting of Cobrelimab (TSR-022; Tesaro), RG7769 (Genentech), MAS825 (Novartis), Sabatrolizumab (MBG453; Novartis), Sym023 (Symphogen), INCAGN2390 (Incyte), LY3321367 (Eli Lilly and Company), BMS-986258 (BMS), SHR-1702 (Jiangsu HengRui), AZD7789 (AstraZeneca), TQB2618 (Chia Tai T ianqing Pharmaceutical Group Co., Ltd.) and NB002 (Neologics Bioscience), BGBA425 (Beigene) and Tim-3 and PD-1 bispecific RO7121661 (Roche). In some embodiments, the triraciclib, PD-1 or PD-L1 inhibitor, and TIM-3 inhibitor are administered on Day 1 of each 21-day cycle, and are administered on Day 7 and On the 14th day, Trirasili was administered again. In some embodiments, the PD-1 or PD-L1 inhibitor comprises the PD-1 inhibitor dotalimab. In some embodiments, the TIM-3 inhibitor comprises cobreliumab. In some embodiments, treracil, dotalimab, and cobrelimab are administered on Day 1 of each 21-day cycle, and again on Days 7 and 14 of each 21-day cycle. Vote with Qu Laxili. In some embodiments, the 21-day cycle is repeated in the absence of disease progression or unacceptable toxicity. In some embodiments, dotalimumab and cobrelimab are administered intravenously within 30 minutes. In some embodiments, dotalimab is administered at a dose of about 500 mg. In some embodiments, cobrezumab is administered at a dose of about 300 mg.

In one embodiment, the other checkpoint inhibitor is a LAG-3 checkpoint inhibitor. In one embodiment, the LAG-3 checkpoint inhibitor is selected from the group consisting of Rilarizumab ( ^OPDUALAG® ; BMS-986016; Bristol-Myers Squibb), GSK2831781 (GlaxoSmithKline), Effiragimo De alpha (IMP321; Prima BioMed), liramumab (LAG525; Novartis), favrizumab (MK-4280; Merck), framilizumab (REGN3767; Regeneron), TSR-033 (Tesaro/ GSK), BI754111 (Boehringer Ingelheim), Sym022 (Symphogen), LBL-007 (Nanjing Leads Biolabs Co., Ltd), IBI110 (Innovent Biologics), IBI323 (Innovent Biologics), INCAGN02385 (Incyte Corporation), AVA021 (A vacta), MGD013 (Macrogenics), RO7247669 (Hoffman-LaRoche), EMB-02 (Shanghai Epimab Biotherapeutics), XmAb841 (Xencor), dual PD-1 and LAG-3 inhibitor tepolizumab (MGD013; MacroGenics), CB213 (Crescendo Biologics) and SNA-03 (Microbio Group) and the dual PD-L1 and LAG-3 inhibitor FS118 (F-Star).

In one embodiment, the other checkpoint inhibitor is a CD73 checkpoint inhibitor. In one embodiment, the CD73 checkpoint inhibitor is selected from the group consisting of HLX23 (Shanghai Henlius Biotech), LY3475070 (Eli Lilly and Company), IPH5301 (Innate Pharma, Astra Zeneca), AK119 (Akesobio Australia Pty Ltd. ), PT199 (Phanes Therapeutics), mupadolizumab (CPI-006; Corvus Pharmaceuticals), Sym024 (Symphogen), olevolumab (MEDI9447; Astra Zeneca), IBI325 (Innovent Biologics), ORIC-533 (Oric Pharmaceuticals), JAB-BX102 (Jacobio Pharmaceuticals), TJ004309 (Tracon Pharmaceuticals), AB680 (Arcus Biosciences), NZV930 (Novartis), BMS-986179 (Bristol Myers Squibb), INCA00186 (Incyte Corporation) and anti-CD73-T GFβ-Trap Dual Functional antibody Darofop alpha (Gilead Sciences).

Non-squamous NSCLC In one aspect, the improved method of treatment described herein is administered to patients with locally advanced or metastatic non-squamous NSCLC in a first-line advanced/metastatic setting, wherein the patient is administered With triracicill, chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic non-squamous NSCLC in the first-line advanced/metastatic setting, wherein the patient is administered triaciclib , PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic non-squamous NSCLC in a second-line advanced/metastatic setting, wherein the patient is administered trela cilium, chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic non-squamous NSCLC in a second-line advanced/metastatic setting, wherein the patient is administered trela Sealy, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic non-squamous NSCLC in a second-line advanced/metastatic setting, wherein the patient is administered trela Sealy, PD-1 or PD-L1 inhibitors and multiple tyrosine kinase (MTK) inhibitors. In some embodiments, the MTK inhibitor is selected from levatinib, stretratinib, or crizotinib. In some embodiments, the patient has a tumor expressing PD-L1. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test.

In one embodiment, CDK4/6 inhibitors and PD-1 or PD-L1 checkpoint inhibitors and other immune checkpoint inhibitors (ICI) (selected from T cell immune receptors with Ig and ITIM domains (TIGIT ) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint inhibitor, or cluster of differentiation 73 (CD73) checkpoint inhibitor ) can be used in combination with many standard-of-care chemotherapy regimens, such as, but not limited to, first-line or second-line treatment regimens for squamous cell NSCLC, such as, but not limited to, single-agent chemotherapy or combination chemotherapy. A non-limiting example of a single agent chemotherapy regimen is 500 mg/ ^m2 IV pemetrexed every 21 days. A non-limiting example of a combination chemotherapy regimen includes Day 1 AUC 5 IV Carboplatin + Day 1 500 mg/ ^m IV Pemetrexed every 21 days for 4 cycles, Day ¹ 75 mg/m IV Cisplatin + 500 mg/m ² IV pemetrexed on day 1 every 21 days for three cycles, 75 mg/m ² IV cisplatin on day 1 + 175 mg/m ² IV Pacific on day 1 every 21 days Paclitaxel, 100 mg/m ² IV cisplatin on day 1 + 1000 mg/m ² IV gemcitabine on days 1, 8, and 15 every 28 days, 60 mg/m ² IV cisplatin on day 1 + every 21 days on days 1 and 15 1000 mg/ ^m2 IV gemcitabine on day 8, 75 mg/ ^m2 IV cisplatin on day 1 + 75 mg/ ^m2 IV docetaxel on day 1 every 21 days, AUC on day 1 6 IV carboplatin + every 21 days Day 175-225 mg/m ² IV paclitaxel on day 1, AUC on day 1 6 IV carboplatin + 90 mg/m ² IV paclitaxel on days 1, 8 and 15 every 28 days, days 1 and 8 every 21 days And 100 mg/m ² IV protein-bound paclitaxel for 15 days + AUC 6 IV carboplatin on day 1, AUC 6 IV carboplatin on day 1 + 75 mg/m ² IV docetaxel on day 1 every 21 days, Day 1 AUC 5 IV carboplatin + 1250 mg/m ² IV gemcitabine on days 1 and 8 every 21 days, 100 mg ^{/m 2 IV cisplatin on day 1 every 28 days + 25 mg/m 2 IV} vinorel every week Vinorelbine, 40 mg/m ² IV cisplatin on day 1 + 25 mg/m ² IV vinorelbine on days 1 and 8 every 21 days or AUC 5 IV carboplatin on day 1 + 30 mg/m 2 IV carboplatin on days 1 and 8 every 21 days mg/m ² IV vinorelbine. \

In one embodiment, the CDK4/6 inhibitor is treracicil. In some embodiments, treracilib is administered less than 4 hours prior to administration of the first-line chemotherapy regimen. In some embodiments, treracicill is administered about one hour or less (eg, about 45 minutes, about 40 minutes, about 35 minutes, or about 30 minutes) prior to the administration of the first-line chemotherapy regimen. In some embodiments, the triracicil is administered intravenously to the patient at about 190 mg/m ² to 280 mg/m ² . In some embodiments, treracilil is administered at about 240 mg/m ² .

In one embodiment, the immune checkpoint inhibitor is a PD-1 inhibitor. In one embodiment, the immune checkpoint inhibitor is a PD-1 inhibitor selected from the group consisting of: nivolumab (OPDIVO ^® ), pembrolizumab (KEYTRUDA ^® ), cimipril anti (LIBTAYO ^® ) and dotalimab (JEMPERLI ^® ). In one embodiment, the PD-1 inhibitor is Nivolumab. In some embodiments, nivolumab is administered with platinum-containing chemotherapy at 360 mg on the same day every 3 weeks for 3 cycles. In some embodiments, nivolumab is administered at 240 mg every 2 weeks or at 480 mg every 4 weeks. In one embodiment, the PD-1 inhibitor is pembrolizumab. In some embodiments, pembrolizumab is administered at 200 mg every 3 weeks or at 400 mg every 6 weeks. In one embodiment, the PD-1 inhibitor is simiprizumab. In some embodiments, cimiprizumab 350 mg is administered as an intravenous infusion over 30 minutes every 3 weeks. In one embodiment, the PD-1 inhibitor is dotalimab. In some embodiments, dotalimab is administered as an intravenous infusion over 30 minutes at 500 mg every 3 weeks (for doses 1-4) and then at 1,000 mg every 6 weeks.

In one embodiment, the immune checkpoint inhibitor is a PD-L1 inhibitor. In one embodiment, the PD-L1 inhibitor is selected from the group consisting of atezolizumab (TECENTRIQ ^® ), avelumab (BAVENCIO ^® ) and durvalumab (IMFINZI ^® ). In one embodiment, the PD-LI inhibitor is atezolizumab. In some embodiments, atezolizumab is administered at 840 mg every 2 weeks, 1200 mg every 3 weeks, or 1680 mg every 4 weeks for up to 1 year. In some embodiments, the PD-L1 inhibitor is avelumab. In some embodiments, avelumab 800 mg is administered as an intravenous infusion over 60 minutes every 2 weeks. In one embodiment, the PD-L1 inhibitor is durvalumab. In some embodiments, administered at 10 mg/kg every 2 weeks or at 1500 mg every 4 weeks (for patients weighing greater than 30 kg) and at 10 mg/kg every 2 weeks (for patients weighing less than 30 kg) with durvalumab.

In one embodiment, the other checkpoint inhibitor is a TIGIT checkpoint inhibitor. In one embodiment, the TIGIT checkpoint inhibitor is selected from the group consisting of vebolizumab (MK-7684; Merck), etigimumab/OMP-313 M32 (OncoMed), tisreliumab Anti-(MTIG7192A/RG-6058; Roche/Genentech), Ospolizumab (BGB-A1217; Beigene), BMS-986207 (BMS), COM902 (Compugen), M6223 (Merck KGaA), Dovanarumab (AB-154; Arcus Biosciences), AZD2936 (AstraZeneca), JS006 (Shanghai Junshi Bioscience), IBI139 (Innovent Biologics), ASP-8374 (Astellas/Potenza), BAT6021 (Bio-Thera Solutions), TAB006 (Shanghai Junshi Bioscience) , Domvanalimab (AB154; Arcus Biosciences), EOS884448 (EOS-448; iTeos), SEA-TGT (Seattle Genetics), mAb-7 (Stanwei Biotech), SHR-1708 (Hengrui Medicine), GS02 (Suzhou Zelgen/Qilu Pharma) , RXI-804 (Rxi Pharmaceuticals), NB6253 (Northern Biologics), ENUM009 (Enumreal Biomedical), CASC-674 (Cascadian Therapeutics), AJUD008 (AJUD Biopharma), AGEN1777 (Agenus, Bristol-Myers Squibb), HLX53 (Squibb) hanghai Henlius Biotech ), BAT6005 (Bio-Thera Solutions), anti-TIGIT/anti-PD-L1 bispecific antibody HLX301 (Shanghai Henlius Biotech) and anti-TIGIT/anti-PD-L1 antibody HB0036 (Shanghai Huaota Biopharmaceutical).

In one embodiment, the other checkpoint inhibitor is a TIM-3 checkpoint inhibitor. In one embodiment, the TIM-3 checkpoint inhibitor is selected from the group consisting of Cobrelimab (TSR-022; Tesaro), RG7769 (Genentech), MAS825 (Novartis), Sabatrolizumab (MBG453; Novartis), Sym023 (Symphogen), INCAGN2390 (Incyte), LY3321367 (Eli Lilly and Company), BMS-986258 (BMS), SHR-1702 (Jiangsu HengRui), AZD7789 (AstraZeneca), TQB2618 (Chia Tai T ianqing Pharmaceutical Group Co., Ltd.) and NB002 (Neologics Bioscience), BGBA425 (Beigene) and Tim-3 and PD-1 bispecific RO7121661 (Roche).

In one embodiment, the other checkpoint inhibitor is a LAG-3 checkpoint inhibitor. In one embodiment, the LAG-3 checkpoint inhibitor is selected from the group consisting of Rilarizumab ( ^OPDUALAG® ; BMS-986016; Bristol-Myers Squibb), GSK2831781 (GlaxoSmithKline), Effiragimo De alpha (IMP321; Prima BioMed), liramumab (LAG525; Novartis), favrizumab (MK-4280; Merck), framilizumab (REGN3767; Regeneron), TSR-033 (Tesaro/ GSK), BI754111 (Boehringer Ingelheim), Sym022 (Symphogen), LBL-007 (Nanjing Leads Biolabs Co., Ltd), IBI110 (Innovent Biologics), IBI323 (Innovent Biologics), INCAGN02385 (Incyte Corporation), AVA021 (A vacta), MGD013 (Macrogenics), RO7247669 (Hoffman-LaRoche), EMB-02 (Shanghai Epimab Biotherapeutics), XmAb841 (Xencor), dual PD-1 and LAG-3 inhibitor tepolizumab (MGD013; MacroGenics), CB213 (Crescendo Biologics) and SNA-03 (Microbio Group) and the dual PD-L1 and LAG-3 inhibitor FS118 (F-Star).

In one embodiment, the other checkpoint inhibitor is a CD73 checkpoint inhibitor. In one embodiment, the CD73 checkpoint inhibitor is selected from the group consisting of HLX23 (Shanghai Henlius Biotech), LY3475070 (Eli Lilly and Company), IPH5301 (Innate Pharma, Astra Zeneca), AK119 (Akesobio Australia Pty Ltd. ), PT199 (Phanes Therapeutics), mupadolizumab (CPI-006; Corvus Pharmaceuticals), Sym024 (Symphogen), olevolumab (MEDI9447; Astra Zeneca), IBI325 (Innovent Biologics), ORIC-533 (Oric Pharmaceuticals), JAB-BX102 (Jacobio Pharmaceuticals), TJ004309 (Tracon Pharmaceuticals), AB680 (Arcus Biosciences), NZV930 (Novartis), BMS-986179 (Bristol Myers Squibb), INCA00186 (Incyte Corporation) and anti-CD73-T GFβ-Trap Dual Functional antibody Darofop alpha (Gilead Sciences).

Triple Negative Breast Cancer Triple negative breast cancer (TNBC) is a highly aggressive subtype of breast cancer that accounts for 15-20% of breast cancer cases and 25% of all breast cancer deaths each year. TNBC is characterized by several aggressive clinicopathologic features, including onset at a younger age; larger high-grade tumors; and a propensity for visceral metastases (Cheang et al., Basal-like breast cancer defined by five biomarkers has superior prognostic value than triple-negative phenotype . Clin Cancer Res. 2008; 14(5): 1368-76; Foulkes et al., Triple-negative breast cancer. N Engl J Med. 2010 Nov 11; 363(20): 1938-48).

Breast cancer is usually classified as TNBC based on localized ER-negative, progesterone receptor (PR)-negative, HER2-negative status, which can be assessed by histological or cytologic hormone receptor immunohistochemistry (IHC) (for estrogen and progesterone receptors). progesterone, defined as <1% nuclei staining) and by IHC [0 or 1+] or in situ hybridization [ratio <2.0] or mean gene copy number <4 signals/nuclei (non-overrepresented for HER2 negative) Assay (according to 2018 American Society of Clinical Oncology and the College of American Pathologists (ASCO CAP) guidelines).

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic triple-negative breast cancer (TNBC) in a first-line advanced/metastatic setting, wherein Trila is administered to the patient cilium, chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic triple-negative breast cancer (TNBC) in a first-line advanced/metastatic setting, wherein Trila is administered to the patient cilium, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic triple-negative breast cancer (TNBC) in a second-line advanced/metastatic setting, wherein the patient is administered Trp Rascilli, chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic triple-negative breast cancer (TNBC) in a second-line advanced/metastatic setting, wherein the patient is administered Trp Rascilli, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one embodiment, CDK4/6 inhibitors and PD-1 or PD-L1 checkpoint inhibitors and other immune checkpoint inhibitors (ICI) (selected from T cell immune receptors with Ig and ITIM domains (TIGIT ) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint inhibitor, or cluster of differentiation 73 (CD73) checkpoint inhibitor ) can be combined with many standard-of-care chemotherapy regimens such as, but not limited to, metastatic TNBC regimens, such as, but not limited to, first-line adjuvant therapy, first-line chemotherapy for metastatic TNBC, and chemotherapy for metastatic TNBC second-line chemotherapy) in combination. Non-limiting examples of first-line adjuvant therapy for metastatic TNBC include: oral administration of 1,000-1,250 mg/m ^capecitabine twice daily on days 1-14, repeated every 3 weeks; 30-minute AUC 6 IV carboplatin repeated every 3 or 4 weeks; 75 mg/ ^m IV cisplatin over 60 minutes on day 1 repeated every 3 weeks; 60-75 mg/m IV on day 1 ² Doxorubicin (repeated cycle every 3 weeks) or 20 mg/ ^m2 doxorubicin intravenously on day 1 (repeated cycle every week); 1.4 mg/ ^m2 intravenously injected on days 1 and 8 Eribulin, repeated every 3 weeks; gemcitabine 800-1,200 mg/ ^m2 IV over 30 minutes on days 1, 8, and 15, repeated every 4 weeks; liposomal 40-50 mg/ ^m2 IV on day 1 Doxorubicin, repeat cycle every 4 weeks; 175 mg/ ^m2 IV paclitaxel over 3 hours on Day 1 (repeat cycle every 3 weeks) or 80 mg/ ^m2 IV paclitaxel over 60 minutes on Day 1 (weekly 25 mg/ ^m2 vinorelbine over 5-10 minutes on day 1, repeated weekly; 260 mg/ ^m2 IV nab-paclitaxel over 30 minutes on day 1 (repeated every 3 weeks ) or 100 mg/m ² IV nab-paclitaxel over 30 minutes on days 1, 8, and 15 (repeated cycle every 4 weeks) or 125 mg/m ² IV albumin over 30 minutes on days 1, 8, and 15 Conjugated paclitaxel (repeated cycle every 4 weeks); Cyclophosphamide 50 mg PO once daily on days 1-21, repeated cycle every 4 weeks; 60-100 mg/m ² IV over 60 minutes on day 1 Docetaxel (repeated cycle every 3 weeks) or 35 mg/ ^m2 IV docetaxel over 60 minutes on Days 1, 8, 15, 22, 29, and 36; 60-90 mg IV on Day 1 Epirubicin/ ^m2 , repeated every 3 weeks; Day 1 40 mg/ ^m2 (max 88 mg) IV ixabepilone over 3 hours, repeated every 3 weeks; Days 1, 15: Day 1 and 840 mg IV atezolizumab over 60 minutes on day 15, followed by: 100 mg/m ² IV nab-paclitaxel on days 1, 8, and 15, repeated every 4 weeks.

In one embodiment, the CDK4/6 inhibitor is treracicil. In some embodiments, treracilib is administered less than 4 hours prior to administration of the first-line chemotherapy regimen. In some embodiments, treracicill is administered about one hour or less (eg, about 45 minutes, about 40 minutes, about 35 minutes, or about 30 minutes) prior to the administration of the first-line chemotherapy regimen. In some embodiments, the triaciclib is administered to the patient intravenously at about 190 mg/m ² to 280 mg/m ² . In some embodiments, treracilil is administered at about 240 mg/m ² .

In one embodiment, the immune checkpoint inhibitor is a PD-1 inhibitor. In one embodiment, the immune checkpoint inhibitor is a PD-1 inhibitor selected from the group consisting of: nivolumab (OPDIVO ^® ), pembrolizumab (KEYTRUDA ^® ), cimipril anti (LIBTAYO ^® ) and dotalimab (JEMPERLI ^® ). In one embodiment, the PD-1 inhibitor is Nivolumab. In some embodiments, nivolumab is administered at 360 mg on the same day every 3 weeks with platinum-containing chemotherapy for 3 cycles. In some embodiments, nivolumab is administered at 240 mg every 2 weeks or at 480 mg every 4 weeks. In one embodiment, the PD-1 inhibitor is pembrolizumab. In some embodiments, pembrolizumab is administered at 200 mg every 3 weeks or at 400 mg every 6 weeks. In one embodiment, the PD-1 inhibitor is simiprizumab. In some embodiments, cimiprizumab 350 mg is administered as an intravenous infusion over 30 minutes every 3 weeks. In one embodiment, the PD-1 inhibitor is dotalimab. In some embodiments, dotalimab is administered as an intravenous infusion over 30 minutes at 500 mg every 3 weeks (for doses 1-4) and then at 1,000 mg every 6 weeks.

In one embodiment, the immune checkpoint inhibitor is a PD-L1 inhibitor. In one embodiment, the PD-L1 inhibitor is selected from the group consisting of atezolizumab (TECENTRIQ ^® ), avelumab (BAVENCIO ^® ) and durvalumab (IMFINZI ^® ). In one embodiment, the PD-LI inhibitor is atezolizumab. In some embodiments, atezolizumab is administered at 840 mg every 2 weeks, 1200 mg every 3 weeks, or 1680 mg every 4 weeks for up to 1 year. In some embodiments, the PD-L1 inhibitor is avelumab. In some embodiments, avelumab 800 mg is administered as an intravenous infusion over 60 minutes every 2 weeks. In one embodiment, the PD-L1 inhibitor is durvalumab. In some embodiments, administered at 10 mg/kg every 2 weeks or at 1500 mg every 4 weeks (for patients weighing greater than 30 kg) and at 10 mg/kg every 2 weeks (for patients weighing less than 30 kg) with durvalumab.

In one embodiment, the other checkpoint inhibitor is a TIGIT checkpoint inhibitor. In one embodiment, the TIGIT checkpoint inhibitor is selected from the group consisting of vebolizumab (MK-7684; Merck), etigimumab/OMP-313 M32 (OncoMed), tisreliumab Anti-(MTIG7192A/RG-6058; Roche/Genentech), Ospolizumab (BGB-A1217; Beigene), BMS-986207 (BMS), COM902 (Compugen), M6223 (Merck KGaA), Dovanarumab (AB-154; Arcus Biosciences), AZD2936 (AstraZeneca), JS006 (Shanghai Junshi Bioscience), IBI139 (Innovent Biologics), ASP-8374 (Astellas/Potenza), BAT6021 (Bio-Thera Solutions), TAB006 (Shanghai Junshi Bioscience) , Domvanalimab (AB154; Arcus Biosciences), EOS884448 (EOS-448; iTeos), SEA-TGT (Seattle Genetics), mAb-7 (Stanwei Biotech), SHR-1708 (Hengrui Medicine), GS02 (Suzhou Zelgen/Qilu Pharma) , RXI-804 (Rxi Pharmaceuticals), NB6253 (Northern Biologics), ENUM009 (Enumreal Biomedical), CASC-674 (Cascadian Therapeutics), AJUD008 (AJUD Biopharma), AGEN1777 (Agenus, Bristol-Myers Squibb), HLX53 (Squibb) hanghai Henlius Biotech ), BAT6005 (Bio-Thera Solutions), anti-TIGIT/anti-PD-L1 bispecific antibody HLX301 (Shanghai Henlius Biotech) and anti-TIGIT/anti-PD-L1 antibody HB0036 (Shanghai Huaota Biopharmaceutical).

In one embodiment, the other checkpoint inhibitor is a TIM-3 checkpoint inhibitor. In one embodiment, the TIM-3 checkpoint inhibitor is selected from the group consisting of Cobrelimab (TSR-022; Tesaro), RG7769 (Genentech), MAS825 (Novartis), Sabatrolizumab (MBG453; Novartis), Sym023 (Symphogen), INCAGN2390 (Incyte), LY3321367 (Eli Lilly and Company), BMS-986258 (BMS), SHR-1702 (Jiangsu HengRui), AZD7789 (AstraZeneca), TQB2618 (Chia Tai T ianqing Pharmaceutical Group Co., Ltd.) and NB002 (Neologics Bioscience), BGBA425 (Beigene) and Tim-3 and PD-1 bispecific RO7121661 (Roche).

In one embodiment, the other checkpoint inhibitor is a LAG-3 checkpoint inhibitor. In one embodiment, the LAG-3 checkpoint inhibitor is selected from the group consisting of Rilarizumab (Opdualag®; BMS-986016; Bristol-Myers Squibb), GSK2831781 (GlaxoSmithKline), Effiragimo De alpha (IMP321; Prima BioMed), liramumab (LAG525; Novartis), favrizumab (MK-4280; Merck), framilizumab (REGN3767; Regeneron), TSR-033 (Tesaro/ GSK), BI754111 (Boehringer Ingelheim), Sym022 (Symphogen), LBL-007 (Nanjing Leads Biolabs), IBI110 (Innovent Biologics), IBI323 (Innovent Biologics), INCAGN02385 (Incyte Corporation), AVA021 (Avacta), M GD013 (Macrogenics) , RO7247669 (Hoffman-LaRoche), EMB-02 (Shanghai Epimab Biotherapeutics), XmAb841 (Xencor), dual PD-1 and LAG-3 inhibitor tepolizumab (MGD013; MacroGenics), CB213 (Crescendo Biologics) and SNA -03 (Microbio Group) and dual PD-L1 and LAG-3 inhibitor FS118 (F-Star).

In one embodiment, the other checkpoint inhibitor is a CD73 checkpoint inhibitor. In one embodiment, the CD73 checkpoint inhibitor is selected from the group consisting of HLX23 (Shanghai Henlius Biotech), LY3475070 (Eli Lilly and Company), IPH5301 (Innate Pharma, Astra Zeneca), AK119 (Akesobio Australia Pty Ltd. ), PT199 (Phanes Therapeutics), mupadolizumab (CPI-006; Corvus Pharmaceuticals), Sym024 (Symphogen), olevolumab (MEDI9447; Astra Zeneca), IBI325 (Innovent Biologics), ORIC-533 (Oric Pharmaceuticals), JAB-BX102 (Jacobio Pharmaceuticals), TJ004309 (Tracon Pharmaceuticals), AB680 (Arcus Biosciences), NZV930 (Novartis), BMS-986179 (Bristol Myers Squibb), INCA00186 (Incyte Corporation) and anti-CD73-T GFβ-Trap Dual Functional antibody Darofop alpha (Gilead Sciences).

Colorectal cancer (CRC) Colorectal cancer (CRC) is one of the common malignancies with high mortality worldwide. The incidence of CRC is increasing day by day, and it is estimated that the number of CRC patients will reach 2.5 million by 2035. CRC is an aggressive cancer whose development and progression involve genetic and environmental factors. The current therapeutic modality for early stage CRC is surgery, followed by radiation therapy and chemotherapy. These commonly used treatments can create several problems, such as their side effects, and often involve the development of drug resistance. To overcome these barriers, other treatment modalities and common therapies aimed at achieving better results are needed. Checkpoint inhibitors can be used in colorectal cancer cells that have tested positive for specific genetic changes, such as microsatellite instability-high (MSI-H) or a type of mismatch repair (MMR) gene change.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic and unresectable colorectal cancer in the first-line advanced/metastatic setting, wherein the patient is administered trachomatis Rascilli, chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1. In some embodiments, the patient has a tumor that is microsatellite instability-high (MSI-H) or mismatch repair-deficient (dMMR)-type as confirmed by an FDA-approved test.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic and unresectable colorectal cancer in the first-line advanced/metastatic setting, wherein the patient is administered trachomatis Rascilli, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1. In some embodiments, the patient has a tumor that is microsatellite instability-high (MSI-H) or mismatch repair-deficient (dMMR)-type as confirmed by an FDA-approved test.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic and unresectable colorectal cancer in a second-line advanced/metastatic setting, wherein the patient is administered Tracicill, chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1. In some embodiments, the patient has a tumor that is microsatellite instability-high (MSI-H) or mismatch repair-deficient (dMMR)-type as confirmed by an FDA-approved test.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic and unresectable colorectal cancer in a second-line advanced/metastatic setting, wherein the patient is administered Tracicill, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1. In some embodiments, the patient has a tumor that is microsatellite instability-high (MSI-H) or mismatch repair-deficient (dMMR)-type as confirmed by an FDA-approved test.

In one embodiment, CDK4/6 inhibitors and PD-1 or PD-L1 checkpoint inhibitors and other immune checkpoint inhibitors (ICI) (selected from T cell immune receptors with Ig and ITIM domains (TIGIT ) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint inhibitor, or cluster of differentiation 73 (CD73) checkpoint inhibitor ) can be combined with many standard-of-care chemotherapy regimens such as (but not limited to) metastatic colorectal cancer treatment regimens such as (but not limited to): postoperative adjuvant chemotherapy for colorectal cancer, colorectal cancer First-line chemotherapy and second-line chemotherapy for colorectal cancer). Non-limiting examples for first-line chemotherapy for colorectal cancer include: FOLFIRI - 180 mg/ ^m2 irinotecan on day 1, 400 mg/ ^m2 folate on day 1 over two hours 400 mg/m ² bolus fluorouracil followed by 2400 mg/m ² to 3000 mg/m ² over 46 hours, continuous infusion; repeat cycle every 2 weeks; Douillard regimen - 180 mg/m ² on day 1 Irinotecan, 200 mg/ ^m2 folate over two hours on days 1 and 2 followed by 400 mg/m2 bolus fluorouracil on days 1 and ² and then 600 mg/ ^m2 over 22 hours; Repeat cycle every 2 weeks; FOLFOX 4 - 85 mg/m ² oxaliplatin on day 1, 400 mg/m ² folate on days 1 and 2 two hours before fluorouracil, 400 on days 1 and 2 mg/ ^m2 bolus fluorouracil followed by 600 mg/ ^m2 over 22 hours; repeat cycle every two weeks; FOLFOX 6 - 100 mg/ ^m2 oxaliplatin on day 1, 400 mg/m2 over two hours on day 1 ² Formyltetrahydrofolate, 400 mg/ ^m2 bolus fluorouracil on day 1 followed by 2400 mg/ ^m2 to 3000 mg/ ^m2 over 46 hours, continuous infusion, repeat cycle every two weeks; modified FOLFOX 6-day 85 mg/m ² oxaliplatin on day 1, 350 mg total folate over two hours on day 1, 400 mg/m ² bolus fluorouracil on day 1 followed by 2400 mg/m ² over 46 hours, Continuous infusion, repeating cycle every two weeks; FOLFOX 7 - 130 mg/m ² oxaliplatin on day 1, 400 mg/m ² folate on day 1 over two hours, 400 mg/m ² on day 1 Fluorouracil bolus followed by 2400 mg/ ^m2 over 46 hours, continuous infusion, repeated every two weeks; modified FOLFOX7 - 85 mg/ ^m2 oxaliplatin on day 1, 200 mg/m2 over two hours on day 1 ² formyltetrahydrofolate, 2400 mg/ ^m2 fluorouracil over 46 hours, continuous infusion, repeated cycle every two weeks; XELOX - 130 mg/ ^m2 oxaliplatin on day 1, twice daily on days 1 to 14 Oral administration of 1000 mg/m ² capecitabine, repeated cycle every 3 weeks; FOLFOXIRI - 165 mg/m ² irinotecan on day 1, 85 mg/m ² oxaliplatin on day 1, oxaliplatin on day 1 400 mg/ ^m2 folate for two hours followed by 3200 mg/ ^m2 fluorouracil over 48 hours, repeated every two weeks.

In one embodiment, the CDK4/6 inhibitor is treracicil. In some embodiments, treracilib is administered less than 4 hours prior to administration of the first-line chemotherapy regimen. In some embodiments, treracicill is administered about one hour or less (eg, about 45 minutes, about 40 minutes, about 35 minutes, or about 30 minutes) prior to the administration of the first-line chemotherapy regimen. In some embodiments, the triracicil is administered intravenously to the patient at about 190 mg/m ² to 280 mg/m ² . In some embodiments, treracilil is administered at about 240 mg/m ² .

In one embodiment, the immune checkpoint inhibitor is a PD-1 inhibitor. In one embodiment, the immune checkpoint inhibitor is a PD-1 inhibitor selected from the group consisting of: nivolumab (OPDIVO ^® ), pembrolizumab (KEYTRUDA ^® ), cimipril anti (LIBTAYO ^® ) and dotalimab (JEMPERLI ^® ). In one embodiment, the PD-1 inhibitor is Nivolumab. In some embodiments, nivolumab is administered with platinum-containing chemotherapy at 360 mg on the same day every 3 weeks for 3 cycles. In some embodiments, nivolumab is administered at 240 mg every 2 weeks or at 480 mg every 4 weeks. In one embodiment, the PD-1 inhibitor is pembrolizumab. In some embodiments, pembrolizumab is administered at 200 mg every 3 weeks or at 400 mg every 6 weeks. In one embodiment, the PD-1 inhibitor is simiprizumab. In some embodiments, cimiprizumab 350 mg is administered as an intravenous infusion over 30 minutes every 3 weeks. In one embodiment, the PD-1 inhibitor is dotalimab. In some embodiments, dotalimab is administered as an intravenous infusion over 30 minutes at 500 mg every 3 weeks (for doses 1-4) and then at 1,000 mg every 6 weeks.

In one embodiment, the immune checkpoint inhibitor is a PD-L1 inhibitor. In one embodiment, the PD-L1 inhibitor is selected from the group consisting of atezolizumab (TECENTRIQ ^® ), avelumab (BAVENCIO ^® ) and durvalumab (IMFINZI ^® ). In one embodiment, the PD-LI inhibitor is atezolizumab. In some embodiments, atezolizumab is administered at 840 mg every 2 weeks, 1200 mg every 3 weeks, or 1680 mg every 4 weeks for up to 1 year. In some embodiments, the PD-L1 inhibitor is avelumab. In some embodiments, avelumab 800 mg is administered as an intravenous infusion over 60 minutes every 2 weeks. In one embodiment, the PD-L1 inhibitor is durvalumab. In some embodiments, administered at 10 mg/kg every 2 weeks or at 1500 mg every 4 weeks (for patients weighing greater than 30 kg) and at 10 mg/kg every 2 weeks (for patients weighing less than 30 kg) with durvalumab.

In one embodiment, the other checkpoint inhibitor is a TIGIT checkpoint inhibitor. In one embodiment, the TIGIT checkpoint inhibitor is selected from the group consisting of vebolizumab (MK-7684; Merck), etigimumab/OMP-313 M32 (OncoMed), tisreliumab Anti-(MTIG7192A/RG-6058; Roche/Genentech), Ospolizumab (BGB-A1217; Beigene), BMS-986207 (BMS), COM902 (Compugen), M6223 (Merck KGaA), Dovanarumab (AB-154; Arcus Biosciences), AZD2936 (AstraZeneca), JS006 (Shanghai Junshi Bioscience), IBI139 (Innovent Biologics), ASP-8374 (Astellas/Potenza), BAT6021 (Bio-Thera Solutions), TAB006 (Shanghai Junshi Bioscience) , Domvanalimab (AB154; Arcus Biosciences), EOS884448 (EOS-448; iTeos), SEA-TGT (Seattle Genetics), mAb-7 (Stanwei Biotech), SHR-1708 (Hengrui Medicine), GS02 (Suzhou Zelgen/Qilu Pharma) , RXI-804 (Rxi Pharmaceuticals), NB6253 (Northern Biologics), ENUM009 (Enumreal Biomedical), CASC-674 (Cascadian Therapeutics), AJUD008 (AJUD Biopharma), AGEN1777 (Agenus, Bristol-Myers Squibb), HLX53 (Squibb) hanghai Henlius Biotech ), BAT6005 (Bio-Thera Solutions), anti-TIGIT/anti-PD-L1 bispecific antibody HLX301 (Shanghai Henlius Biotech) and anti-TIGIT/anti-PD-L1 antibody HB0036 (Shanghai Huaota Biopharmaceutical).

In one embodiment, the other checkpoint inhibitor is a TIM-3 checkpoint inhibitor. In one embodiment, the TIM-3 checkpoint inhibitor is selected from the group consisting of Cobrelimab (TSR-022; Tesaro), RG7769 (Genentech), MAS825 (Novartis), Sabatrolizumab (MBG453; Novartis), Sym023 (Symphogen), INCAGN2390 (Incyte), LY3321367 (Eli Lilly and Company), BMS-986258 (BMS), SHR-1702 (Jiangsu HengRui), AZD7789 (AstraZeneca), TQB2618 (Chia Tai T ianqing Pharmaceutical Group Co., Ltd.) and NB002 (Neologics Bioscience), BGBA425 (Beigene) and Tim-3 and PD-1 bispecific RO7121661 (Roche).

In one embodiment, the other checkpoint inhibitor is a LAG-3 checkpoint inhibitor. In one embodiment, the LAG-3 checkpoint inhibitor is selected from the group consisting of Rilarimab (Opdualag®; BMS-986016; Bristol-Myers Squibb), GSK2831781 (GlaxoSmithKline), Effiragimo De alpha (IMP321; Prima BioMed), liramumab (LAG525; Novartis), favrizumab (MK-4280; Merck), framilizumab (REGN3767; Regeneron), TSR-033 (Tesaro/ GSK), BI754111 (Boehringer Ingelheim), Sym022 (Symphogen), LBL-007 (Nanjing Leads Biolabs), IBI110 (Innovent Biologics), IBI323 (Innovent Biologics), INCAGN02385 (Incyte Corporation), AVA021 (Avacta), M GD013 (Macrogenics) , RO7247669 (Hoffman-LaRoche), EMB-02 (Shanghai Epimab Biotherapeutics), XmAb841 (Xencor), dual PD-1 and LAG-3 inhibitor tepolizumab (MGD013; MacroGenics), CB213 (Crescendo Biologics) and SNA -03 (Microbio Group) and dual PD-L1 and LAG-3 inhibitor FS118 (F-Star).

In one embodiment, the other checkpoint inhibitor is a CD73 checkpoint inhibitor. In one embodiment, the CD73 checkpoint inhibitor is selected from the group consisting of HLX23 (Shanghai Henlius Biotech), LY3475070 (Eli Lilly and Company), IPH5301 (Innate Pharma, Astra Zeneca), AK119 (Akesobio Australia Pty Ltd. ), PT199 (Phanes Therapeutics), mupadolizumab (CPI-006; Corvus Pharmaceuticals), Sym024 (Symphogen), olevolumab (MEDI9447; Astra Zeneca), IBI325 (Innovent Biologics), ORIC-533 (Oric Pharmaceuticals), JAB-BX102 (Jacobio Pharmaceuticals), TJ004309 (Tracon Pharmaceuticals), AB680 (Arcus Biosciences), NZV930 (Novartis), BMS-986179 (Bristol Myers Squibb), INCA00186 (Incyte Corporation) and anti-CD73-T GFβ-Trap Dual Functional antibody Darofop alpha (Gilead Sciences).

Metastatic urothelial carcinoma Metastatic urothelial (transitional cell) carcinoma (mUC) is the major histological type of bladder cancer in the United States and Europe, which accounts for 90% of all bladder cancers. Bladder cancer is the 6th most common cancer in men and the 17th most common cancer in women worldwide. Bladder cancer is the most common malignant tumor involving the urinary system. Bladder cancer can be classified as non-muscle-invasive, muscle-invasive, or metastatic. Approximately 25% of patients will have muscle invasive disease that presents or subsequently develops metastases. Systemic chemotherapy is the standard modality for the initial treatment of patients with inoperable locally advanced or metastatic urothelial malignancies. Despite the high initial response rate, the median survival with multiagent chemotherapy is approximately 15 months.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic urothelial carcinoma (mUC) in a first-line advanced/metastatic setting, wherein the patient is administered Trp Rascilli, chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic urothelial carcinoma (mUC) in a first-line advanced/metastatic setting, wherein the patient is administered Trp Rascilli, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic urothelial carcinoma (mUC) in a second-line advanced/metastatic setting, wherein the patient is administered Tracicill, chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one aspect, the improved method of treatment described herein is administered to a patient with locally advanced or metastatic urothelial carcinoma (mUC) in a second-line advanced/metastatic setting, wherein the patient is administered Tracicill, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1.

In one embodiment, CDK4/6 inhibitors and PD-1 or PD-L1 checkpoint inhibitors and other immune checkpoint inhibitors (ICI) (selected from T cell immune receptors with Ig and ITIM domains (TIGIT ) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint inhibitor, or cluster of differentiation 73 (CD73) checkpoint inhibitor ) can be combined with many standard-of-care chemotherapy regimens, such as (but not limited to) metastatic urothelial carcinoma treatment regimens, such as (but not limited to): postoperative adjuvant intravesical chemotherapy for metastatic urothelial carcinoma , first-line chemotherapy for metastatic urothelial carcinoma and second-line chemotherapy for metastatic urothelial carcinoma) in combination. Non-limiting examples of first-line chemotherapy for metastatic urothelial carcinoma include: 1000 mg/ ^m2 gemcitabine on days 1, 8, and 15 + 70 mg/ ^m2 cisplatin on day 2, repeated every 28 days and Up to 6 cycles; 30 mg/m ² IV methotrexate on days 1, 15, and 22 + 3 mg/m ² IV vinblastine on days 2, 15, and 22 + 30 mg/m ² IV on day 2 Doxorubicin + 70 mg/m ² IV cisplatin on day 2, repeated every 28 days and a total of 6 cycles; paclitaxel (80 mg/m ² on days 1 and 8 before gemcitabine and cisplatin), gemcitabine ( 1000 mg/m ² on days 1 and 8) and cisplatin (70 mg/m ² on day 1), repeated every 21 days for a maximum of 6 cycles; and the above dose-dense regimen administered with doses of growth factor stimulators .

In one embodiment, the CDK4/6 inhibitor is treracicil. In some embodiments, treracilib is administered less than 4 hours prior to administration of the first-line chemotherapy regimen. In some embodiments, treracicill is administered about one hour or less (eg, about 45 minutes, about 40 minutes, about 35 minutes, or about 30 minutes) prior to administration of the first-line chemotherapy regimen. In some embodiments, the triracicil is administered intravenously to the patient at about 190 mg/m ² to 280 mg/m ² . In some embodiments, treracilil is administered at about 240 mg/m ² .

In one embodiment, the immune checkpoint inhibitor is a PD-1 inhibitor. In one embodiment, the immune checkpoint inhibitor is a PD-1 inhibitor selected from the group consisting of: nivolumab (OPDIVO ^® ), pembrolizumab (KEYTRUDA ^® ), cimipril anti (LIBTAYO ^® ) and dotalimab (JEMPERLI ^® ). In one embodiment, the PD-1 inhibitor is Nivolumab. In some embodiments, nivolumab is administered with platinum-containing chemotherapy at 360 mg on the same day every 3 weeks for 3 cycles. In some embodiments, nivolumab is administered at 240 mg every 2 weeks or at 480 mg every 4 weeks. In one embodiment, the PD-1 inhibitor is pembrolizumab. In some embodiments, pembrolizumab is administered at 200 mg every 3 weeks or at 400 mg every 6 weeks. In one embodiment, the PD-1 inhibitor is simiprizumab. In some embodiments, cimiprizumab 350 mg is administered as an intravenous infusion over 30 minutes every 3 weeks. In one embodiment, the PD-1 inhibitor is dotalimab. In some embodiments, dotalimab is administered as an intravenous infusion over 30 minutes at 500 mg every 3 weeks (for doses 1-4) and then at 1,000 mg every 6 weeks.

In one embodiment, the immune checkpoint inhibitor is a PD-L1 inhibitor. In one embodiment, the PD-L1 inhibitor is selected from the group consisting of atezolizumab (TECENTRIQ ^® ), avelumab (BAVENCIO ^® ) and durvalumab (IMFINZI ^® ). In one embodiment, the PD-LI inhibitor is atezolizumab. In some embodiments, atezolizumab is administered at 840 mg every 2 weeks, 1200 mg every 3 weeks, or 1680 mg every 4 weeks for up to 1 year. In some embodiments, the PD-L1 inhibitor is avelumab. In some embodiments, avelumab 800 mg is administered as an intravenous infusion over 60 minutes every 2 weeks. In one embodiment, the PD-L1 inhibitor is durvalumab. In some embodiments, administered at 10 mg/kg every 2 weeks or at 1500 mg every 4 weeks (for patients weighing greater than 30 kg) and at 10 mg/kg every 2 weeks (for patients weighing less than 30 kg) with durvalumab.

In one embodiment, the other checkpoint inhibitor is a TIGIT checkpoint inhibitor. In one embodiment, the TIGIT checkpoint inhibitor is selected from the group consisting of vebolizumab (MK-7684; Merck), etigimumab/OMP-313 M32 (OncoMed), tisreliumab Anti-(MTIG7192A/RG-6058; Roche/Genentech), Ospolizumab (BGB-A1217; Beigene), BMS-986207 (BMS), COM902 (Compugen), M6223 (Merck KGaA), Dovanarumab (AB-154; Arcus Biosciences), AZD2936 (AstraZeneca), JS006 (Shanghai Junshi Bioscience), IBI139 (Innovent Biologics), ASP-8374 (Astellas/Potenza), BAT6021 (Bio-Thera Solutions), TAB006 (Shanghai Junshi Bioscience) , Domvanalimab (AB154; Arcus Biosciences), EOS884448 (EOS-448; iTeos), SEA-TGT (Seattle Genetics), mAb-7 (Stanwei Biotech), SHR-1708 (Hengrui Medicine), GS02 (Suzhou Zelgen/Qilu Pharma) , RXI-804 (Rxi Pharmaceuticals), NB6253 (Northern Biologics), ENUM009 (Enumreal Biomedical), CASC-674 (Cascadian Therapeutics), AJUD008 (AJUD Biopharma), AGEN1777 (Agenus, Bristol-Myers Squibb), HLX53 (Squibb) hanghai Henlius Biotech ), BAT6005 (Bio-Thera Solutions), anti-TIGIT/anti-PD-L1 bispecific antibody HLX301 (Shanghai Henlius Biotech) and anti-TIGIT/anti-PD-L1 antibody HB0036 (Shanghai Huaota Biopharmaceutical).

In one embodiment, the other checkpoint inhibitor is a TIM-3 checkpoint inhibitor. In one embodiment, the TIM-3 checkpoint inhibitor is selected from the group consisting of Cobrelimab (TSR-022; Tesaro), RG7769 (Genentech), MAS825 (Novartis), Sabatrolizumab (MBG453; Novartis), Sym023 (Symphogen), INCAGN2390 (Incyte), LY3321367 (Eli Lilly and Company), BMS-986258 (BMS), SHR-1702 (Jiangsu HengRui), AZD7789 (AstraZeneca), TQB2618 (Chia Tai T ianqing Pharmaceutical Group Co., Ltd.) and NB002 (Neologics Bioscience), BGBA425 (Beigene) and Tim-3 and PD-1 bispecific RO7121661 (Roche).

In one embodiment, the other checkpoint inhibitor is a LAG-3 checkpoint inhibitor. In one embodiment, the LAG-3 checkpoint inhibitor is selected from the group consisting of Rilarizumab (Opdualag®; BMS-986016; Bristol-Myers Squibb), GSK2831781 (GlaxoSmithKline), Effiragimo De alpha (IMP321; Prima BioMed), liramumab (LAG525; Novartis), favrizumab (MK-4280; Merck), framilizumab (REGN3767; Regeneron), TSR-033 (Tesaro/ GSK), BI754111 (Boehringer Ingelheim), Sym022 (Symphogen), LBL-007 (Nanjing Leads Biolabs), IBI110 (Innovent Biologics), IBI323 (Innovent Biologics), INCAGN02385 (Incyte Corporation), AVA021 (Avacta), M GD013 (Macrogenics) , RO7247669 (Hoffman-LaRoche), EMB-02 (Shanghai Epimab Biotherapeutics), XmAb841 (Xencor), dual PD-1 and LAG-3 inhibitor tepolizumab (MGD013; MacroGenics), CB213 (Crescendo Biologics) and SNA -03 (Microbio Group) and dual PD-L1 and LAG-3 inhibitor FS118 (F-Star).

In one embodiment, the other checkpoint inhibitor is a CD73 checkpoint inhibitor. In one embodiment, the CD73 checkpoint inhibitor is selected from the group consisting of HLX23 (Shanghai Henlius Biotech), LY3475070 (Eli Lilly and Company), IPH5301 (Innate Pharma, Astra Zeneca), AK119 (Akesobio Australia Pty Ltd. ), PT199 (Phanes Therapeutics), mupadolizumab (CPI-006; Corvus Pharmaceuticals), Sym024 (Symphogen), olevolumab (MEDI9447; Astra Zeneca), IBI325 (Innovent Biologics), ORIC-533 (Oric Pharmaceuticals), JAB-BX102 (Jacobio Pharmaceuticals), TJ004309 (Tracon Pharmaceuticals), AB680 (Arcus Biosciences), NZV930 (Novartis), BMS-986179 (Bristol Myers Squibb), INCA00186 (Incyte Corporation) and anti-CD73-T GFβ-Trap Dual Functional antibody Darofop alpha (Gilead Sciences).

Solid Tumors In one aspect, the improved method of treatment described herein is administered to a patient with a locally advanced or metastatic solid tumor in the first-line advanced/metastatic setting, wherein the patient is administered treacillib, Chemotherapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the solid tumor is selected from cervical cancer, head and neck squamous cell carcinoma (SCCHN), cutaneous squamous cell carcinoma (cSCC), Merkel cell carcinoma, basal cell carcinoma, small cell lung cancer (SCLC), melanoma Malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, high microsatellite instability or mismatch repair deficient cancer, endometrial cancer, high tumor mutation burden (TMB-H) cancer, gastric cancer, gastroesophageal cancer Carcinoma of the junction, adenocarcinoma of the esophagus, or cancer of the esophagus. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test.

In one aspect, the improved methods of treatment described herein are administered to patients with locally advanced or metastatic solid tumors in the first-line advanced/metastatic setting, wherein the patients are administered treracillib, PD- 1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the solid tumor is selected from cervical cancer, head and neck squamous cell carcinoma (SCCHN), cutaneous squamous cell carcinoma (cSCC), Merkel cell carcinoma, basal cell carcinoma, small cell lung cancer (SCLC), melanoma Malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, high microsatellite instability or mismatch repair deficient cancer, endometrial cancer, high tumor mutation burden (TMB-H) cancer, gastric cancer, gastroesophageal cancer Carcinoma of the junction, adenocarcinoma of the esophagus, or cancer of the esophagus. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test.

In one aspect, the improved method of treatment described herein is administered to patients with locally advanced or metastatic solid tumors in a second-line advanced/metastatic setting, wherein the patient is administered treracilil, chemo Therapeutic agents, PD-1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the solid tumor is selected from cervical cancer, head and neck squamous cell carcinoma (SCCHN), cutaneous squamous cell carcinoma (cSCC), Merkel cell carcinoma, basal cell carcinoma, small cell lung cancer (SCLC), melanoma Malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, high microsatellite instability or mismatch repair deficient cancer, endometrial cancer, high tumor mutation burden (TMB-H) cancer, gastric cancer, gastroesophageal cancer Carcinoma of the junction, adenocarcinoma of the esophagus, or cancer of the esophagus. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test.

In one aspect, the improved methods of treatment described herein are administered to patients with locally advanced or metastatic solid tumors in a second-line advanced/metastatic setting, wherein the patients are administered triaciclib, PD -1 or PD-L1 inhibitors and other immune checkpoint inhibitors selected from TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors or CD73 inhibitors. In some embodiments, the patient has a tumor expressing PD-L1. In some embodiments, the solid tumor is selected from cervical cancer, head and neck squamous cell carcinoma (SCCHN), cutaneous squamous cell carcinoma (cSCC), Merkel cell carcinoma, basal cell carcinoma, small cell lung cancer (SCLC), melanoma Malignant pleural mesothelioma, renal cell carcinoma, hepatocellular carcinoma, high microsatellite instability or mismatch repair deficient cancer, endometrial cancer, high tumor mutation burden (TMB-H) cancer, gastric cancer, gastroesophageal cancer Carcinoma of the junction, adenocarcinoma of the esophagus, or cancer of the esophagus. In some embodiments, the patient has a tumor expressing PD-L1, as determined by an FDA approved test or a CE mark test.

In one embodiment, CDK4/6 inhibitors and PD-1 or PD-L1 checkpoint inhibitors and other immune checkpoint inhibitors (ICI) (selected from T cell immune receptors with Ig and ITIM domains (TIGIT ) checkpoint inhibitor, T cell immunoglobulin mucin-3 (TIM-3) checkpoint inhibitor, lymphocyte activation gene 3 (LAG-3) checkpoint inhibitor, or cluster of differentiation 73 (CD73) checkpoint inhibitor ) combinations can be used in conjunction with many standard-of-care chemotherapy regimens used to treat solid tumors.

In one embodiment, the CDK4/6 inhibitor is treracicil. In some embodiments, treracilib is administered less than 4 hours prior to administration of the first-line chemotherapy regimen. In some embodiments, treracicill is administered about one hour or less (eg, about 45 minutes, about 40 minutes, about 35 minutes, or about 30 minutes) prior to the administration of the first-line chemotherapy regimen. In some embodiments, the triracicil is administered intravenously to the patient at about 190 mg/m ² to 280 mg/m ² . In some embodiments, treracilil is administered at about 240 mg/m ² .

In one embodiment, the immune checkpoint inhibitor is a PD-1 inhibitor. In one embodiment, the immune checkpoint inhibitor is a PD-1 inhibitor selected from the group consisting of: nivolumab (OPDIVO ^® ), pembrolizumab (KEYTRUDA ^® ), cimipril anti (LIBTAYO ^® ) and dotalimab (JEMPERLI ^® ). In one embodiment, the PD-1 inhibitor is Nivolumab. In some embodiments, nivolumab is administered with platinum-containing chemotherapy at 360 mg on the same day every 3 weeks for 3 cycles. In some embodiments, nivolumab is administered at 240 mg every 2 weeks or at 480 mg every 4 weeks. In one embodiment, the PD-1 inhibitor is pembrolizumab. In some embodiments, pembrolizumab is administered at 200 mg every 3 weeks or at 400 mg every 6 weeks. In one embodiment, the PD-1 inhibitor is simiprizumab. In some embodiments, cimiprizumab 350 mg is administered as an intravenous infusion over 30 minutes every 3 weeks. In one embodiment, the PD-1 inhibitor is dotalimab. In some embodiments, dotalimab is administered as an intravenous infusion over 30 minutes at 500 mg every 3 weeks (for doses 1-4) and then at 1,000 mg every 6 weeks.

In one embodiment, the immune checkpoint inhibitor is a PD-L1 inhibitor. In one embodiment, the PD-L1 inhibitor is selected from the group consisting of atezolizumab (TECENTRIQ ^® ), avelumab (BAVENCIO ^® ) and durvalumab (IMFINZI ^® ). In one embodiment, the PD-LI inhibitor is atezolizumab. In some embodiments, atezolizumab is administered at 840 mg every 2 weeks, 1200 mg every 3 weeks, or 1680 mg every 4 weeks for up to 1 year. In some embodiments, the PD-L1 inhibitor is avelumab. In some embodiments, avelumab 800 mg is administered as an intravenous infusion over 60 minutes every 2 weeks. In one embodiment, the PD-L1 inhibitor is durvalumab. In some embodiments, administered at 10 mg/kg every 2 weeks or at 1500 mg every 4 weeks (for patients weighing greater than 30 kg) and at 10 mg/kg every 2 weeks (for patients weighing less than 30 kg) with durvalumab.

In one embodiment, the other checkpoint inhibitor is a TIGIT checkpoint inhibitor. In one embodiment, the TIGIT checkpoint inhibitor is selected from the group consisting of vebolizumab (MK-7684; Merck), etigimumab/OMP-313 M32 (OncoMed), tisreliumab Anti-(MTIG7192A/RG-6058; Roche/Genentech), Ospolizumab (BGB-A1217; Beigene), BMS-986207 (BMS), COM902 (Compugen), M6223 (Merck KGaA), Dovanarumab (AB-154; Arcus Biosciences), AZD2936 (AstraZeneca), JS006 (Shanghai Junshi Bioscience), IBI139 (Innovent Biologics), ASP-8374 (Astellas/Potenza), BAT6021 (Bio-Thera Solutions), TAB006 (Shanghai Junshi Bioscience) , Domvanalimab (AB154; Arcus Biosciences), EOS884448 (EOS-448; iTeos), SEA-TGT (Seattle Genetics), mAb-7 (Stanwei Biotech), SHR-1708 (Hengrui Medicine), GS02 (Suzhou Zelgen/Qilu Pharma) , RXI-804 (Rxi Pharmaceuticals), NB6253 (Northern Biologics), ENUM009 (Enumreal Biomedical), CASC-674 (Cascadian Therapeutics), AJUD008 (AJUD Biopharma), AGEN1777 (Agenus, Bristol-Myers Squibb), HLX53 (Squibb) hanghai Henlius Biotech ), BAT6005 (Bio-Thera Solutions), anti-TIGIT/anti-PD-L1 bispecific antibody HLX301 (Shanghai Henlius Biotech) and anti-TIGIT/anti-PD-L1 antibody HB0036 (Shanghai Huaota Biopharmaceutical).

In one embodiment, the other checkpoint inhibitor is a TIM-3 checkpoint inhibitor. In one embodiment, the TIM-3 checkpoint inhibitor is selected from the group consisting of Cobrelimab (TSR-022; Tesaro), RG7769 (Genentech), MAS825 (Novartis), Sabatrolizumab (MBG453; Novartis), Sym023 (Symphogen), INCAGN2390 (Incyte), LY3321367 (Eli Lilly and Company), BMS-986258 (BMS), SHR-1702 (Jiangsu HengRui), AZD7789 (AstraZeneca), TQB2618 (Chia Tai T ianqing Pharmaceutical Group Co., Ltd.) and NB002 (Neologics Bioscience), BGBA425 (Beigene) and Tim-3 and PD-1 bispecific RO7121661 (Roche).

In one embodiment, the other checkpoint inhibitor is a LAG-3 checkpoint inhibitor. In one embodiment, the LAG-3 checkpoint inhibitor is selected from the group consisting of Rilarizumab (Opdualag®; BMS-986016; Bristol-Myers Squibb), GSK2831781 (GlaxoSmithKline), Effiragimo De alpha (IMP321; Prima BioMed), liramumab (LAG525; Novartis), favrizumab (MK-4280; Merck), framilizumab (REGN3767; Regeneron), TSR-033 (Tesaro/ GSK), BI754111 (Boehringer Ingelheim), Sym022 (Symphogen), LBL-007 (Nanjing Leads Biolabs), IBI110 (Innovent Biologics), IBI323 (Innovent Biologics), INCAGN02385 (Incyte Corporation), AVA021 (Avacta), M GD013 (Macrogenics) , RO7247669 (Hoffman-LaRoche), EMB-02 (Shanghai Epimab Biotherapeutics), XmAb841 (Xencor), dual PD-1 and LAG-3 inhibitor tepolizumab (MGD013; MacroGenics), CB213 (Crescendo Biologics) and SNA -03 (Microbio Group) and dual PD-L1 and LAG-3 inhibitor FS118 (F-Star).

In one embodiment, the other checkpoint inhibitor is a CD73 checkpoint inhibitor. In one embodiment, the CD73 checkpoint inhibitor is selected from the group consisting of HLX23 (Shanghai Henlius Biotech), LY3475070 (Eli Lilly and Company), IPH5301 (Innate Pharma, Astra Zeneca), AK119 (Akesobio Australia Pty Ltd. ), PT199 (Phanes Therapeutics), mupadolizumab (CPI-006; Corvus Pharmaceuticals), Sym024 (Symphogen), olevolumab (MEDI9447; Astra Zeneca), IBI325 (Innovent Biologics), ORIC-533 (Oric Pharmaceuticals), JAB-BX102 (Jacobio Pharmaceuticals), TJ004309 (Tracon Pharmaceuticals), AB680 (Arcus Biosciences), NZV930 (Novartis), BMS-986179 (Bristol Myers Squibb), INCA00186 (Incyte Corporation) and anti-CD73-T GFβ-Trap Dual Functional antibody Darofop alpha (Gilead Sciences).

Pharmaceutical Compositions and Dosage Forms An effective amount of an active compound described herein, or a salt, isotopic analog or prodrug thereof, for use in the methods described herein can be administered to a subject using any suitable means to achieve the desired therapeutic result. The amount and timing of active compound administered will, of course, depend on the subject treated, the instructions of the supervising medical professional, the schedule of exposure, the mode of administration, the pharmacokinetic properties of the particular active compound, and the judgment of the prescribing physician. Accordingly, due to variability among hosts, the dosages given below are guidelines and the physician may adjust the dosage of the active compounds to achieve what the physician deems appropriate for the host's treatment. In considering the desired degree of treatment, a physician can balance various factors, such as the age and weight of the host, the presence or absence of pre-existing diseases, and the presence or absence of other diseases. Typical administered doses of CDK4/6 inhibitors such as Compound I have been previously described in WO 2016/126889, the entire content of which is incorporated herein.

Pharmaceutical compositions can be administered in therapeutically effective amounts by any desired mode of administration, but are usually administered by intravenous injection or infusion. In alternative embodiments, an effective amount of the compound or pharmaceutically acceptable salt is delivered with a pharmaceutically acceptable carrier for oral delivery. By way of more general non-limiting example, the pharmaceutical composition is suitable for the following modes of administration: oral (including buccal and sublingual), rectal, nasal, topical, dermal, pulmonary, vaginal or parenteral (including intramuscular, intraarterial, intrathecal, subcutaneous and intravenous), injection, inhalation or spray, intraaortic, intracranial, subdermal, intraperitoneal, subcutaneous or other pharmaceutically acceptable carriers Investment method.

The therapeutically effective dose of any active compound described herein will be determined by the healthcare practitioner having regard to the condition, size and age of the patient and the route of delivery. In one non-limiting example, dosages of about 0.1 mg/kg to about 200 mg/kg are therapeutically effective, wherein all weights are based on the weight of the active compound, including the use of salts. In some embodiments, the dosage may provide up to about 10 nM, 50 nM, 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, 600 nM, 700 nM, 800 nM, 900 nM, 1 μM, 5 μM , 10 μM, 20 μM, 30 μM, or 40 μM of the amount of compound required for a serum concentration of active compound.

In certain embodiments, dosage forms of pharmaceutical compositions contain from about 0.1 mg to about 2000 mg, from about 10 mg to about 1000 mg, from about 100 mg to about 800 mg, or from about 200 mg to about 600 mg of the active compound in a unit dosage form And optionally from about 0.1 mg to about 2000 mg, from about 10 mg to about 1000 mg, from about 100 mg to about 800 mg, or from about 200 mg to about 600 mg of the other active agent. Examples of dosage forms contain at least 0.01, 0.05, 0.1, 1, 5, 10, 15, 20, 25, 50, 100, 200, 250, 300, 400, 500, 600, 700 or 750 mg of active compound or a salt thereof. Pharmaceutical compositions may also contain the active compound and another active agent in a molar ratio that achieves the desired result.

An effective amount of a disclosed compound, or a salt thereof, can be administered based on the patient's weight, size, or age. For example, a therapeutic amount may, for example, range from about 0.01 mg/kg body weight to about 250 mg/kg body weight or from about 0.1 mg/kg to about 10 mg/kg in at least one dose. As many doses as necessary to reduce and/or alleviate and/or cure the condition in question can be administered to the patient. When desired, formulations can be prepared with enteric coatings suitable for sustained or controlled release administration of the active ingredient.

In certain embodiments, the dose is between about 0.01-500 mg/kg patient body weight, for example about 0.01 mg/kg, about 0.05 mg/kg, about 0.1 mg/kg, about 0.5 mg/kg, about 1 mg /kg, about 1.5 mg/kg, about 2 mg/kg, about 2.5 mg/kg, about 3 mg/kg, about 3.5 mg/kg, about 4 mg/kg, about 4.5 mg/kg, about 5 mg/kg , about 10 mg/kg, about 15 mg/kg, about 20 mg/kg, about 25 mg/kg, about 30 mg/kg, about 35 mg/kg, about 40 mg/kg, about 45 mg/kg, about 50 mg/kg, about 55 mg/kg, about 60 mg/kg, about 65 mg/kg, about 70 mg/kg, about 75 mg/kg, about 80 mg/kg, about 85 mg/kg, about 90 mg /kg, about 95 mg/kg, about 100 mg/kg, about 105 mg/kg, about 110 mg/kg, about 115 mg/kg, about 120 mg/kg, about 125 mg/kg, about 130 mg/kg , about 135 mg/kg, about 140 mg/kg, about 145 mg/kg, about 150 mg/kg, about 155 mg/kg, about 160 mg/kg, about 165 mg/kg, about 170 mg/kg, about 175 mg/kg, about 180 mg/kg, about 185 mg/kg, about 190 mg/kg, about 195 mg/kg, 200 mg/kg, about 205 mg/kg, about 210 mg/kg, about 215 mg/kg kg, about 220 mg/kg, about 225 mg/kg, about 230 mg/kg, about 235 mg/kg, about 240 mg/kg, about 245 mg/kg, about 250 mg/kg, about 255 mg/kg, About 260 mg/kg, About 265 mg/kg, About 270 mg/kg, About 275 mg/kg, About 280 mg/kg, About 285 mg/kg, About 290 mg/kg, About 295 mg/kg, 300 mg /kg, about 305 mg/kg, about 310 mg/kg, about 315 mg/kg, about 320 mg/kg, about 325 mg/kg, about 330 mg/kg, about 335 mg/kg, about 340 mg/kg , about 345 mg/kg, about 350 mg/kg, about 355 mg/kg, about 360 mg/kg, about 365 mg/kg, about 370 mg/kg, about 375 mg/kg, about 380 mg/kg, about 385 mg/kg, about 390 mg/kg, about 395 mg/kg, 400 mg/kg, about 405 mg/kg, about 410 mg/kg, about 415 mg/kg, about 420 mg/kg, about 425 mg/kg kg, about 430 mg/kg, about 435 mg/kg, about 440 mg/kg, about 445 mg/kg, about 450 mg/kg, about 455 mg/kg, about 460 mg/kg, about 465 mg/kg, About 470 mg/kg, about 475 mg/kg, about 480 mg/kg, about 485 mg/kg, about 490 mg/kg, about 495 mg/kg, or about 500 mg/kg.

In one embodiment, the CDK4/6 inhibitor administered is administered at a dose of about 180 mg/ ^m2 to about 280 mg/ ^m2 . In one embodiment, at about 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275 or about 280 mg/m ² cast treracilide. In one embodiment, treracilil is administered at a dose of about 200 mg/m ² . In one embodiment, treracilil is administered at a dose of about 240 mg/m ² .

Pharmaceutical formulations are preferably in unit dosage form. In such form, the preparation is subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.

In certain embodiments, the administered compound is administered as a pharmaceutically acceptable salt. Non-limiting examples of pharmaceutically acceptable salts include: acetate, adipate, alginate, ascorbate, aspartate, besylate, benzoate, bisulfate, borate , butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, lauryl sulfate, ethanesulfonate, fumarate, glucoheptose Glycerophosphate, Hemisulfate, Heptanoate, Hexanoate, Hydrobromide, Hydrochloride, Hydroiodide, 2-Hydroxy-ethanesulfonate, Lactobionate, Lactate, Lauryl salt, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, Palmitate, Pamoate, Pectinate, Persulfate, 3-Phenylpropionate, Phosphate, Picrate, Pivalate, Propionate, Stearate, Succinate salt, sulfate, tartrate, thiocyanate, tosylate, undecanoate and valerate. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium as well as non-toxic ammonium, quaternary ammonium, and amine cations including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine , trimethylamine, triethylamine and ethylamine.

The compounds disclosed herein or used as described herein may be administered orally, topically, parenterally, by inhalation or spray in dosage unit formulations containing conventional pharmaceutically acceptable carriers. , sublingually, via implant (including ocular implant), transdermally, via buccal administration, rectally, in the form of an ophthalmic solution, injection (including ocular injection), intravenous, intramuscular, Inhalation, intraaortic, intracranial, subdermal, intraperitoneal, subcutaneous, nasal, sublingual or rectal or by other means. For ocular delivery, the compound may be administered as desired, for example, via immediate or controlled release, or via an ocular device: intravitreal, intrastromal, intracameral, sub-tenon ), subretinal, retrobulbar, peribulbar, suprachoroidal, conjunctival, subconjunctival, episcleral, periocular, transscleral, retrobulbar, retroscleral, pericorneal, or lacrimal duct injection or via mucus, mucin, or mucous membranes barrier.

According to the methods disclosed herein, oral administration can be in any desired form, such as solid, gel or liquid (including solution, suspension or emulsion). In some embodiments, the compounds or salts are administered by inhalation, intravenously or intramuscularly as a liposomal suspension. When administered by inhalation, the active compound or salt may be in the form of a plurality of particles of any desired particle size, such as from about 0.01, 0.1, or 0.5 microns to about 5, 10, 20 or more microns, and optionally from about 1 micron to about 2 microns. In the form of solid particles or liquid droplets. Compounds as disclosed in the present invention have shown good pharmacokinetic and pharmacodynamic properties, eg when administered by oral or intravenous routes.

Pharmaceutical formulations may include an active compound described herein, or a pharmaceutically acceptable salt thereof, in any pharmaceutically acceptable carrier. Water is sometimes an optional carrier for water soluble compounds or salts if it is desired to use solutions. For water soluble compounds or salts, organic vehicles such as glycerol, propylene glycol, polyethylene glycol or mixtures thereof may be suitable. In the latter case, the organic vehicle can contain substantial amounts of water. The solution in any case may then be sterilized in a suitable manner known to those skilled in the art, for example by filtration through a 0.22 micron filter. Following sterilization, the solution can be dispensed into appropriate receptacles (eg, depyrogenated glass vials). Dispense by aseptic method as appropriate. The sterilized closure can then be placed on the vial and the contents of the vial can be lyophilized if desired.

Carriers comprise excipients and diluents and must be of sufficiently high purity and sufficiently low toxicity to render them suitable for administration to the patient to be treated. The carrier can be inert or it can possess medicinal benefits of its own. Combination compounds are employed in amounts of carrier sufficient to provide a practical amount of administered material per unit dose of compound.

Types of carriers include (but not limited to) binders, buffers, colorants, diluents, disintegrants, emulsifiers, flavoring agents, slippery agents, lubricants, preservatives, stabilizers, surfactants, tablets and wetting agents. Some carriers may be listed as more than one type, for example, vegetable oil may serve as a lubricant in some formulations and a diluent in others. Exemplary pharmaceutically acceptable carriers include sugar, starch, cellulose, powdered tragacanth, malt, gelatin, talc and vegetable oils. Optional active agents can be included in the pharmaceutical compositions which do not substantially interfere with the activity of the compounds of the invention.

Besides the active compounds or their salts, the pharmaceutical formulations may also contain other additives (eg pH adjusting additives). In particular, useful pH adjusting agents include acids such as hydrochloric acid, bases or buffers such as sodium lactate, sodium acetate, sodium phosphate, sodium citrate, sodium borate or sodium gluconate. Additionally, the formulations may contain antimicrobial preservatives. Useful antimicrobial preservatives include methylparaben, propylparaben and benzyl alcohol. Antimicrobial preservatives are typically used when the formulations are placed in vials designed for multiple-dose applications. The pharmaceutical formulations described herein can be lyophilized using techniques well known in the art.

For oral administration, pharmaceutical compositions can take the form of solutions, suspensions, tablets, pills, capsules, powders, and the like. Tablets containing various excipients (such as sodium citrate, calcium carbonate, and calcium phosphate) can be combined with various disintegrants (such as starches (such as potato or tapioca starch) and certain complex silicates), with binders (such as polyvinylpyrrolidone, sucrose, gelatin and acacia). In addition, lubricating agents such as magnesium stearate, sodium lauryl sulfate, and talc are often very useful for tableting purposes. Solid compositions of a similar type can be used as fillers in soft and hard-filled gelatin capsules. Materials in this context also include lactose (milk sugar) and high molecular weight polyethylene glycols. When it is desired to administer an aqueous suspension and/or elixir orally, the compound of the main substance disclosed in the present invention can be mixed with various sweeteners, flavoring agents, coloring agents, emulsifying agents and/or suspending agents and diluents (such as water, ethanol, propylene glycol, glycerin, and combinations thereof).

In yet another embodiment of the subject matter described herein, there is provided an injectable, stable, sterile formulation comprising an active compound as described herein, or a salt thereof, in unit dosage form in a hermetically sealed container. The compound or salt is provided as a lyophilizate which can be reconstituted with a suitable pharmaceutically acceptable carrier to form a liquid formulation suitable for injection into a host. When the compound or salt is substantially soluble in water, the compound or salt can be emulsified in an aqueous carrier with a sufficient amount of a physiologically acceptable emulsifier. Especially useful emulsifiers include phosphatidylcholine and lecithin.

Other embodiments provided herein comprise liposomal formulations of the active compounds disclosed herein. Techniques for forming liposome suspensions are well known in the art. When the compound is a water-soluble salt, the compound can be incorporated into lipid vesicles using conventional liposome technology. In this case, due to the water solubility of the active compound, the active compound can be substantially entrapped within the hydrophilic center or core of the liposome. The lipid layer employed may have any conventional composition and may or may not contain cholesterol. Where the active compound of interest is insoluble in water, again using conventional liposome formation techniques, the salt can be substantially entrapped within the hydrophobic lipid bilayer forming the liposome structure. In either case, the size of the liposomes produced can be reduced, eg, through the use of standard sonication and homogenization techniques. Liposome formulations including the active compounds disclosed herein can be lyophilized to produce a lyophilizate which can be reconstituted with a pharmaceutically acceptable carrier such as water to reconstitute a liposomal suspension.

Pharmaceutical formulations suitable for administration by inhalation in aerosol form are also provided. Such formulations include a solution or suspension of a desired compound or salt thereof described herein or a plurality of solid particles of the compound or salt. The desired formulation can be placed in the chamber and nebulized. Atomization can be achieved by compressed air or by ultrasonic energy to form a plurality of liquid droplets or solid particles including compounds or salts. Liquid droplets or solid particles can, for example, have a particle size in the range of about 0.5 microns to about 10 microns, and optionally about 0.5 microns to about 5 microns. In one embodiment, the solid particles provide controlled release through the use of degradable polymers. Solid particles may be obtained by treating a solid compound or a salt thereof in any suitable manner known in the art, for example by micronization. Optionally, the size of the solid particles or liquid droplets can be from about 1 micron to about 2 microns. In this case, a commercial nebulizer can be utilized for this purpose. The compounds can be administered via an aerosol suspension of respirable particles in the manner set forth in US Patent No. 5,628,984, the disclosure of which is incorporated herein by reference in its entirety.

Also provided are pharmaceutical formulations that allow for the controlled release of the compounds described herein, including through the use of degradable polymers as known in the art.

When pharmaceutical formulations suitable for aerosol administration are in liquid form, the formulations can include a water-soluble active compound in a carrier, including water. A surfactant may be present which, upon nebulization, sufficiently lowers the surface tension of the formulation to form droplets in the desired size range.

As used herein, the term "pharmaceutically acceptable salts" refers to those salts which, within the scope of sound medical judgment, are suitable for contact with a host (such as a human host) without undue toxicity, irritation, allergic reaction and and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended application; and where possible, zwitterionic forms of the compounds of the subject matter disclosed in this invention.

Accordingly, the term "salts" refers to the relatively nontoxic, inorganic and organic acid addition salts of the compounds disclosed herein. Such salts can be prepared during the final isolation and purification of the compounds or by separately reacting the purified compound in its free base form with a suitable organic or inorganic acid and isolating the salt thus formed. Basic compounds can form many different salts with various inorganic and organic acids. The acid addition salts of basic compounds are prepared by contacting the free base form with a sufficient amount of the desired acid to produce the salt in the conventional manner. The free base form can be regenerated by contacting the salt form with a base and isolating the free base in conventional manner. The free base forms and their respective salt forms can differ by certain physical properties, such as solubility in polar solvents. Pharmaceutically acceptable base addition salts can be formed with metals or amines such as alkali and alkaline earth metal hydroxides or organic amines. Examples of metals used as cations include, but are not limited to, sodium, potassium, magnesium, calcium, and the like. Examples of suitable amines include, but are not limited to, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine, and procaine. Caine (procaine). The base addition salts of acidic compounds are prepared by contacting the free acid form with a sufficient amount of the desired base to produce the salt in the conventional manner. The free acid form can be regenerated by contacting the salt form with an acid and isolating the free acid in conventional manner. Minor differences between the free acid form and its respective salt form can lie in certain physical properties, such as solubility in polar solvents.

Salts can be prepared from inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, phosphorus, and the like, such as sulfates, pyrosulfates, bisulfates, sulfurous acids Salt, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate , borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthenate, methanesulfonate, Glucoheptonate, lactobionate, laurylsulfonate and isethionate, and the like. Salts can also be prepared from organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, and the like. Representative salts include acetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate Salt, Mandelate, Benzoate, Chlorobenzoate, Methylbenzoate, Dinitrobenzoate, Phthalate, Benzenesulfonate, Toluenesulfonate, Benzene Glycolacetates, citrates, lactates, maleates, tartrates, methanesulfonates and the like. Pharmaceutically acceptable salts may include cations based on alkali and alkaline earth metals such as sodium, lithium, potassium, calcium, magnesium, and the like; as well as non-toxic ammonium, quaternary ammonium, and amine cations, including but not limited to ammonium , tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. Salts of amino acids are also contemplated, such as arginate, gluconate, galacturonate, and the like. See, eg, Berge et al., J. Pharm. Sci., 1977, 66, 1-19, which is incorporated herein by reference.

The claimed invention is further illustrated by the following non-limiting examples. Other aspects and embodiments of the present invention will become apparent to those skilled in the art from the foregoing disclosure and the following experimental illustrations, which are included by way of illustration and not limitation, and with reference to the accompanying drawings.

Example 1. Effect of Triracicill ( Alone or in Combination with LAG-3 ) on IFN- γ Production of Depleted T Cells After Restimulation . Splenocytes from MBP-chased mice were stimulated with WT-MBP or APL-MBP for 72 hours to generate non-exhausted and exhausted cells, respectively. T cells were purified and rested, then re-stimulated in the presence of increasing concentrations of treracilil or vehicle control +/- anti-LAG-3 (10 ug/mL). Cell supernatants were collected at the end of the culture to evaluate IFN-g production by ELISA. Data are presented as mean +/- SEM. The results are shown in Figure 1.

Example 2. In vivo tumor studies using chemotherapy /ICI inhibitors +/- treracilil. Nine-week-old female C57BL/6 (C57BL/6NCrl) and BALB/c mice were subcutaneously implanted with 5× ¹⁰⁵ MC3822 or CT26 American Type Culture Collection (American Type Culture Collection, ATCC) tumor cells (provided by Cell line supplied by Charles River Laboratories). Two to three weeks after tumor injection and before the start of treatment (Day 1 of the study), animals with individual tumor volumes of 80 ^mm3 to 120 ^mm3 were sorted into an appropriate number of treatment groups with an average tumor volume of 100 mm ³ . Triracillib (100 mg/kg), oxaliplatin (10 mg/kg; Fresenius Kabi USA, lot number: 8760467A01) or 5-fluorouracil (5-FU; 75 mg/kg) was administered weekly intraperitoneally (IP). , Fresenius Kabi USA, lot number: 6113613) and last for 3 weeks. Anti-PD-L1 (BioXCell, catalog number: BE0101, clonal 10F.9G2, 100 µg/animal, IP) or anti-programmed death-1 (PD-1; BioXCell, catalog number: BE0146, clonal RMP1- 14, 5mg/kg, IP). Tumors were measured twice a week using calipers. Each animal was euthanized when the tumor volume reached the endpoint of 1000 mm3 or on the last day of the study. A partial response (PR) indicates a tumor volume < 50% of its Day 1 volume in three consecutive measurements during the course of the study and > 13.5 mm ³ in at least one of the three measurements. A complete response (CR) indicates a tumor volume < 13.5 mm ³ in three consecutive measurements during the course of the study. Animals were scored for PR or CR events only once during the study, and CR was only scored when both PR and CR criteria were met. The results for MC38 treated mice are shown in Figures 2A-4B and the results for CT26 treated mice are shown in Figures 5A-5B. The results showed that the combination of treracicill and chemotherapy + anti-PD-1 enhanced survival and inhibited tumor growth. Results have been published previously (Anne Y Lai et al., J Immunother Cancer 2020; 8: e000847).

Example 3. Transient G1 arrest alters the proportion of T cell subsets within tumors to benefit effector T cell function. Tumors and spleens were harvested from OP-treated and TOP-treated MC38 mice on days 5 and 9 after the induction+maintenance (IM) treatment regimen. Mouse tumor samples were dissociated using the gentleMACS Protocol Tumor Dissociation Kit (Miltenyi Biotech; Cat#: 130-096-730) according to the manufacturer's instructions. Single cell suspensions were then stained using the LIVE/DEAD Aqua Fixable Dead Cell Staining Kit (Life Technologies) and the crystallizable fragment receptor was blocked using TruStain FcX (Biolegend), followed by staining with an antibody against a cell surface marker: CD8+ T cells (CD45+ CD3+ CD11B-CD8+ CD4-), CD4+ T cells (CD45+ CD3+ CD11B-CD8-CD4+), TREG (CD45+ CD3+ CD11B-CD4+ CD25+ Foxp3+), MMDSC (CD1 1B+ CD3-LY6C+ LY6G-), GMDSC ( CD11b+ CD3- Ly6C+ Ly6G+) and macrophages (CD11b+ CD3- Ly6C-Ly6G-). The Treg proportions of total tumor or splenic CD4+ T cells treated with vehicle, oxaliplatin/PD-1 or triaciclib/oxaliplatin/PD-1 at days 5 and 9 are shown in FIG. 6 . The ratio of CD8+ T cells to Tregs in the CD45+ population (% CD8+ T cells/% Tregs) (n=5-8 tumors analyzed/treatment group and time point) is shown in FIG. 7 . The results showed that the addition of treracilide reduced Tregs and resulted in a higher CD8:Treg ratio, and that treracilide promoted CD8 T cell activation. For FoxP3 staining, cells were permeabilized using transcription factor fixation/permeabilization buffer (eBioscience) and incubated with anti-FoxP3 antibody. The spleen was processed into a single cell suspension, lysed with ammonium chloride-potassium buffer to remove RBCs and stained with antibodies: activated CD8+ T cells (CD8+ CD4- CD69+), activated CD4+ T cells (CD4+ CD8- CD69+) and Treg ( CD4+ CD25+ FoxP3+). The proportion of activated (%CD69+) cells among CD8+ T cells is shown in FIG. 8 . Dead cells were excluded by propidium iodide staining. Antibody clones and supplier information are listed in the online Supplementary Methods. Data were collected on a FACSCanto II (BD Biosciences) and analyzed using FlowJo software (Tree Star). Results have been published previously (Anne Y Lai et al., J Immunother Cancer 2020; 8: e000847).

Example 4. CDK4/6 inhibition augments anti-tumor immunity induced by anti- PD-1 antibodies. MC38 and CT26 cells were injected subcutaneously into 6-8 wk C57BL/6 or Balb/c female mice, respectively. From the indicated time point, the vehicle control, CDK4/6 inhibitor (traciclib) was used alone or together with PD-1 antibody to treat until the end of the experiment using the intermittent dosing schedule of 3 days of taking medicine and 4 days of stopping medicine. PD-1 antibody was administered via IP injection at 200 µg/mouse 3 times a week (Monday, Wednesday and Friday). Tumor volume was monitored every 2–3 days. The tumor growth curves of MC38 treated with CDK4/6 inhibitors or PD-1 antibodies (alone or in combination) are shown in FIG. 9 . C57BL/6 mice with naive or KP tumors were sacrificed and total splenocytes were harvested. Spleen was digested with collagenase D (Roche) and DNase I (Roche) at 37°C for 30 min, followed by incubation with 1×ACS lysis buffer (Biolegend) to lyse red blood cells. Total splenocytes collected were stained with fluorescent dye-conjugated cell surface markers CD3, CD4, CD8, and CD25 to isolate different T cell subsets, including conventional T cell subpopulations, using a BD FACSAria II SORP cell sorter (BD Bioscience). Cells Tconv (CD3+CD4+CD25−), Treg (CD3+CD4+CD25+) and CD8+ (CD3+CD8+). Dead cells were excluded by staining with DAPI (4',6-diamidino-2-phenylindole). Sorted cells were cultured in 96-well plates pre-coated with CD3 antibody (eBioscience) and treated with treracilib in the presence of CD28 (eBioscience). Cells were harvested 3 days after culture and the cytokine production of IFNγ and IL-2 was determined by intracellular staining and analyzed on a BD LSR Fortessa (BD Bioscience). At the end of treatment (day 17), mice were sacrificed and TILs were isolated from tumors for interleukin analysis against IL-2 from CD4+ T cells (left panel) and IFNγ from CD8+ T cells (right panel). (*p<0.001). Results are shown in Tables 10A-10B. This example demonstrates that Tracicill works synergistically with anti-PD-1 to enhance IL-2 production by CD4+ T cells, and that Tracicill alone enhances IFN-γ production in CD8+ T cells and synergizes with anti-PD-1. Results have been published previously (Deng J, Wang ES, Jenkins RW et al., CDK4/6 Inhibition Augments Antitumor Immunity by Enhancing T-cell Activation. Cancer Discov. 2018 Feb;8(2):216-233).

Example 5. Evaluation of Triraciclib and the combination of α-PD-1 and α-TIGIT in the MMTV-PyMT triple negative breast cancer model . 5×10 ⁵ MMTV-PyMT tumor cells were subcutaneously implanted into the fourth inguinal mammary fat pad of 9-week-old female BALB/c mice. One week after tumor cell injection and before the start of treatment (Day 0 of the study), animals were divided into 8 groups according to treatment (N=8/group). Tumor size was between 80-120 ^mm3 . The treatment combination consisted of: 1. Vehicle (citrate buffer) 2. Tracicill (100 mg/kg qwk) 3. Anti-TIGIT (10 mg/kg 2x qwk) 4. Anti-PD-1 (5 mg/kg 2x qwk) 5. Anti-TIGIT (10 mg/kg 2x qwk) + anti-PD-1 (5 mg/kg 2x qwk) 6. Treracilil (100 mg/kg qwk) + anti-TIGIT (10 mg/kg 2x qwk ) 7. Treracilil (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk) 8. Treracillib (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk ) + anti-TIGIT (10 mg/kg 2x qwk) Triracillib (100 mg/kg) administered on day 1 of every week (qwk). Checkpoint inhibitors were administered twice a week (2x qwk) on Day 1 (following treracilib administration) and Day 4 at the concentrations listed above. Treatment lasted 6 weeks.

Triraciclib administered in combination with anti-PD-1 antibody and/or anti-TIGIT antibody reduced overall tumor growth ( FIG. 11A ) and fold change ( FIG. 11B ) in mice xenografted with MMTV-PyMT tumor cells. This could translate into an increase in overall survival (Fig. 11C). Individual tumor growth curves for each treatment group were also plotted separately (FIGS. 11D-11K).

Example 6. Evaluation of Triraciclib and the combination of α -PD-1 and α -TIGIT in the CT26 colorectal cancer model . 5×10 ⁵ CT26 tumor cells were subcutaneously implanted into the axilla of 9-week-old female BALB/c mice. Two separate experiments were run to compare results when dosing was initiated 7 or 10 days after tumor cell injection. Before treatment initiation (day 0 of the study), animals were divided into 8 groups (N=8/group) according to treatment. Tumor size was between 80-120 ^mm3 . The treatment combination for experiments that started treatment after 7 days included: 1. Vehicle (citrate buffer) 2. Treracilil (100 mg/kg qwk) 3. Anti-TIGIT (10 mg/kg 2x qwk) 4. Anti-PD-1 (5 mg/kg 2x qwk) 5. Anti-TIGIT (10 mg/kg 2x qwk) + Anti-PD-1 (5 mg/kg 2x qwk) 6. Treracilil (100 mg/kg qwk) + Anti-TIGIT (10 mg/kg 2x qwk) 7. Treracilil (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk) 8. Treracilil (100 mg/kg qwk) + anti-PD -1 (5 mg/kg 2x qwk) + Anti-TIGIT (10 mg/kg 2x qwk) Triracillib (100 mg/kg) was administered every week (qwk) on day 1 (day 7 after tumor cell injection). Checkpoint inhibitors were administered twice a week (2x qwk) on Day 1 (following treracilib administration) and Day 4 at the concentrations listed above. Drug treatment was discontinued on day 63.

Tracilil administered in combination with anti-PD-1 antibody and/or anti-TIGIT antibody on day 7 reduced overall tumor growth ( FIG. 12A ) and fold change ( FIG. 12B ) in mice xenografted with CT26 tumor cells, This result translated into an increase in overall survival (Figure 12C). Individual tumor growth curves for each treatment group were also plotted separately (FIGS. 12D-12K).

The experimental treatment combination for the treatment started after 10 days consisted of: 1. Vehicle (citrate buffer) 2. Triracilide (100 mg/kg qwk) 3. Anti-TIGIT (10 mg/kg 2x qwk) 4. Treracilil (100 mg/kg qwk) + Anti-TIGIT (10 mg/kg 2x qwk) 5. Treracilil (100 mg/kg qwk) + Anti-PD-1 (5 mg/kg 2x qwk) 6. Treracilil (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk) + anti-TIGIT (10 mg/kg 2x qwk) Triracilil (100 mg/kg) was administered on the first day of every week (qwk) (10th day after tumor cell injection). Checkpoint inhibitors were administered twice a week (2x qwk) on Day 1 (following treracilib administration) and Day 4 at the concentrations listed above. Treatment lasted 6 weeks.

Treracilil administered in combination with anti-PD-1 antibody and/or anti-TIGIT antibody on day 10 reduced tumor growth (fold change) in mice xenografted with CT26 tumor cells (FIG. 13A), which results could be translated is an increase in overall survival (Figure 13B). Individual tumor growth curves for each treatment group were also plotted separately (FIGS. 13C-13H).

Comparing the two experiments it is evident that the efficacy of the combination comprising anti-TIGIT therapy depends on tumor size and that delayed treatment with anti-TIGIT therapy can produce negative results (Figure 14A-C).

Example 7. Evaluation of Triraciclib and α -PD-1 in combination with α -TIM3 or α -LAG3 in the CT26 colorectal cancer model . 5×10 ⁵ CT26 tumor cells were subcutaneously implanted into the axilla of 9-week-old female BALB/c mice. One week after tumor cell injection and before treatment initiation (day 0 of the study), animals were divided into 12 groups (N=8/group) according to treatment. Tumor size ranged from 80-120 ^mm3 . The treatment combination consisted of: 1. Vehicle (citrate buffer) 2. Tracicill (100 mg/kg qwk) 3. Anti-PD-1 (5 mg/kg 2x qwk) 4. Anti-Lag3 (10 mg/kg 2x qwk) 5. Anti-Tim3 (5 mg/kg 2x qwk) 6. Anti-PD-1 (5 mg/kg 2x qwk) + anti-Lag3 (10 mg/kg 2x qwk) 7. Anti-PD-1 (5 mg/kg kg 2x qwk) + anti-Tim3 (5 mg/kg 2x qwk) 8. Treracillib (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk) 9. Treracelib (100 mg/kg qwk) + anti-Lag3 (10 mg/kg 2x qwk) 10. Treracilil (100 mg/kg qwk) + anti-Tim3 (5 mg/kg 2x qwk) 11. Treracilil (100 mg/kg qwk) + anti PD-1 (5 mg/kg 2x qwk) + anti-Lag3 (10 mg/kg 2x qwk) 12. Treracilil (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk) + anti-Tim3 (5 mg/kg 2x qwk) Triracillib (100 mg/kg) was administered on day 1 of every week (qwk). Checkpoint inhibitors were administered twice a week (2x qwk) on Day 1 (following treracilib administration) and Day 4 at the concentrations listed above. Treatment lasted 6 weeks.

Treracilil administered in combination with anti-PD-1 antibody and/or anti-Lag3 or anti-Tim3 antibody on day 7 reduced tumor growth (fold change) in mice xenografted with CT26 tumor cells (Figures 15A-15C) and increased survival (Fig. 15D-E). α-PD-1 + α-LAG3 median TTE was 55.5 days compared to >70 days with the addition of triraciclib (Fig. 15D). Median time to endpoint (TTE) for α-PD-1 + α-TIM3 was not reached with or without treracilib ( FIG. 15E ). Individual tumor growth curves for each treatment group were also plotted separately (FIGS. 15F-15Q).

Example 8. Evaluation of Triraciclib and the combination of α -PD-1 and α -TIGIT in the CT26 colorectal cancer model . 5×10 ⁵ CT26 tumor cells were subcutaneously implanted into the axilla of 9-week-old female BALB/c mice. One week after tumor cell injection and before the start of treatment (Day 0 of the study), animals were divided into 8 groups according to treatment (N=8/group). Tumor size ranged from 80-120 ^mm3 . The treatment combination consisted of: 1. Vehicle (citrate buffer) 2. Tracicill (100 mg/kg qwk) 3. Anti-TIGIT (10 mg/kg 2x qwk) 4. Anti-PD-1 (5 mg/kg 2x qwk) 5. Anti-TIGIT (10 mg/kg 2x qwk) + anti-PD-1 (5 mg/kg 2x qwk) 6. Treracilil (100 mg/kg qwk) + anti-TIGIT (10 mg/kg 2x qwk ) 7. Treracilil (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk) 8. Treracillib (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk ) + Anti-TIGIT (10 mg/kg 2x qwk) Triracilil (100 mg/kg) was administered weekly (qwk) on day 1 (day 7 after tumor cell injection). Checkpoint inhibitors were administered twice a week (2x qwk) on Day 1 (following treracilib administration) and Day 4 at the concentrations listed above. Treatment lasted 6 weeks.

Triracillib administered in combination with anti-PD-1 antibody and/or anti-TIGIT antibody on day 7 reduced tumor volume ( FIG. 16A ) and prolonged overall survival ( FIG. 16B ), where the combination of tramacil and anti-PD1 antibody The combination minimized tumor volume.

Example 9. Evaluation of Triraciclib and the combination of α -PD-1 and α -TIGIT in the AT3-OVA breast cancer model . 5×10 ⁵ AT3-OVA tumor cells were subcutaneously implanted into the fourth inguinal mammary fat pad of 9-week-old female C57BL/6 mice. One week after tumor cell injection and before the start of treatment (Day 0 of the study), animals were divided into 8 groups according to treatment (N=8/group). Tumor size was between 80-120 ^mm3 . The treatment combination consisted of: 1. Vehicle (citrate buffer) 2. Tracicill (100 mg/kg qwk) 3. Anti-TIGIT (10 mg/kg 2x qwk) 4. Anti-PD-1 (5 mg/kg 2x qwk) 5. Anti-TIGIT (10 mg/kg 2x qwk) + anti-PD-1 (5 mg/kg 2x qwk) 6. Treracilil (100 mg/kg qwk) + anti-TIGIT (10 mg/kg 2x qwk ) 7. Treracilil (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk) 8. Treracillib (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk ) + Anti-TIGIT (10 mg/kg 2x qwk) Triracilil (100 mg/kg) was administered weekly (qwk) on day 1 (day 7 after tumor cell injection). Checkpoint inhibitors were administered twice a week (2x qwk) on Day 1 (following treracilib administration) and Day 4 at the concentrations listed above. Treatment lasted 6 weeks.

Tracilil administered in combination with anti-PD-1 antibody and/or anti-TIGIT antibody on day 7 reduced tumor volume (Fig. 17A) and prolonged overall survival (Fig. 17B). The combination of antibodies minimized tumor volume. The combination of treracicill with anti-PD-1 antibody and anti-TIGIT antibody maximized overall survival.

Example 10. Evaluation of Triracilib and the combination of α -PD-1 and α -TIGIT in the S2WTP3 breast cancer model . 5×10 ⁵ S2WTP3 tumor cells were subcutaneously implanted into the fourth inguinal mammary fat pad of 9-week-old female BALB/c mice. One week after tumor cell injection and before the start of treatment (Day 0 of the study), animals were divided into 8 groups according to treatment (N=8/group). Tumor size was between 80-120 ^mm3 . The treatment combination consisted of: 1. Vehicle (citrate buffer) 2. Tracicill (100 mg/kg qwk) 3. Anti-TIGIT (10 mg/kg 2x qwk) 4. Anti-PD-1 (5 mg/kg 2x qwk) 5. Anti-TIGIT (10 mg/kg 2x qwk) + anti-PD-1 (5 mg/kg 2x qwk) 6. Treracilil (100 mg/kg qwk) + anti-TIGIT (10 mg/kg 2x qwk ) 7. Treracilil (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk) 8. Treracillib (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk ) + Anti-TIGIT (10 mg/kg 2x qwk) Triracilil (100 mg/kg) was administered weekly (qwk) on day 1 (day 7 after tumor cell injection). Checkpoint inhibitors were administered twice a week (2x qwk) on Day 1 (following treracilib administration) and Day 4 at the concentrations listed above. Treatment lasted 6 weeks.

Tracilil administered in combination with anti-PD-1 antibody and/or anti-TIGIT antibody on day 7 reduced tumor volume (Figure 18), with the combination of treracillib and anti-PD1 antibody maximally reducing tumor volume .

Example 11. Evaluation of Triraciclib and the Combination of α -PD-1 and α -CD73 in the CT-26 Colorectal Cancer Model 5×10 ⁵ CT26 tumor cells were implanted subcutaneously in 6- to 8-week-old female BALB/c in the armpits of mice. One week after tumor cell injection and before the start of treatment (Day 0 of the study), animals were divided into 8 groups according to treatment (N=8/group). Tumor size ranged from 80-120 ^mm3 . Therapeutic combination consisted of: 1. Vehicle (Citrate Buffer + rIgG2a Isotype Control) 2. Tracillib (100 mg/kg qwk) + rIgG2a Isotype Control (10 mg/kg 2x qwk) 3. Anti-CD73 (5 mg /kg 2x qwk) + vehicle + rIgG2a isotype control (10 mg/kg 2x qwk) 4. Anti-PD-1 (5 mg/kg 2x qwk) + vehicle + rIgG2a isotype control (10 mg/kg 2x qwk) 5 . Triracillib (100 mg/kg qwk) + anti-CD73 (5 mg/kg 2x qwk) + rIgG2a isotype control (10 mg/kg 2x qwk) 6. Triracillib (100 mg/kg qwk) + anti-PD- 1 (5 mg/kg 2x qwk) + rIgG2a isotype control (10 mg/kg 2x qwk) 7. Vehicle + anti-PD-1 (5 mg/kg 2x qwk) + anti-CD73 (5 mg/kg 2x qwk) 8 . Triracilib (100 mg/kg qwk) + anti-PD-1 (5 mg/kg 2x qwk) + anti-CD73 (5 mg/kg 2x qwk) every week (qwk) on day 1 (day 7 after tumor cell injection day) Triracillib (100 mg/kg) was administered. InVivoMAb anti-mouse CD73 clonal TY/23 (BioXcell) and inVivoMAb anti-mouse PD-1 were administered weekly (2x qwk) on Day 1 (following Triracilil administration) and Day 4 at the concentrations listed above Clones RMP1-14 (BioXcell) twice. Treatment lasted 6 weeks. Table 1. Summary of Tumor Growth and Survival by Treatment Groups The effect of IRI was improved by the addition of treracilide Model Group Group tumor growth survival rate CT26 αCD73 Trila + αCD73 0.72 0.39 S2WTP3 αPD-1 Trila + αPD-1 0.004 0.02 CT26 α TIGIT Trila + αTIGIT 0.21 0.04 AT3-OVA α TIGIT Trila + αTIGIT 0.007 0.002 CT26 αLAG3 Trila + αLAG3 0.03 0.13 Synergistic effect of treracilide and IRI combination Model Group Group tumor growth survival rate CT26 αCD73 + αPD-1 Trila + αCD73 + αPD-1 0.2 0.09 S2WTP3 αTIGIT + αPD-1 Trila + αTIGIT + αPD-1 0.09 0.06 CT26 αTIM3 + αPD-1 Trila + αTIM3 + αPD-1 1 0.56 CT26 αLAG3 + αPD-1 Trila + αLAG3 + αPD-1 0.006 0.08 * p-values in italics are significant (p-value < 0.05). IRI, inhibitory receptor immunotherapy.

Tracilil administered in combination with anti-PD-1 antibody and/or anti-CD73 antibody on day 7 reduced tumor growth (tumor volume) in mice xenografted with CT26 tumor cells ( FIG. 20A ), which results could be translated is an increase in overall survival (FIG. 20B).

Based on an additive model, anti-PD-1 treatment had a significant effect on tumor growth at D19 (2.56 e-11) when modulated by anti-CD73 and treracilil. Similarly, treracicill had a significant effect (0.0339) on tumor growth at D19 when modulating anti-CD73 and anti-PD-1. No significant interaction/synergy effects of any drug combination on tumor growth were identified at D17 or D19.

Based on an additive model, anti-PD-1 treatment had a significant effect (1.37e-10) on survival at D19 when modulating anti-CD73 and treracilil. Similarly, anti-CD73 had a significant effect (0.0455) on survival at D19 when adjusted for triracicill and anti-PD-1. No significant interaction/synergy effect on survival was identified for any drug combination at D17 or D19.

Figure 1 shows a line graph showing on the x-axis the concentration of the test compound (traracilil alone or tramacil+anti-LAG-3) measured in nanomolar. The y-axis shows the concentration of IFN-γ measured in pg/ml. Figure 2A shows a line graph illustrating tumor growth. Tumor growth was measured in MC38 mice treated with chemotherapy/ICI±traciclib combination therapy (n=10-15/treatment group). The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 2B shows a line graph of survival curves. The overall survival of MC38 mice treated with oxaliplatin/PD-1 ± treracilil combination therapy was measured (n=10-15/treatment group). The x-axis is days of treatment measured in days and the y-axis is percent survival. Figure 3A shows a line graph illustrating tumor growth. Tumor growth was measured in MC38 mice treated with oxaliplatin/PD-1 ± treracilil combination therapy (n=10-15/treatment group). The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 3B shows a line graph of survival curves. The overall survival of MC38 mice treated with oxaliplatin/PD-1 ± treracilil combination therapy was measured (n=10-15/treatment group). The x-axis is days of treatment measured in days and the y-axis is percent survival. Figure 4A shows a line graph illustrating tumor growth. Tumor growth was measured in MC38 mice treated with 5-FU/PD-L1±traciclib combination therapy (n=10-15/treatment group). The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . The right panel shows Figure 4B showing a line graph of survival curves. The overall survival of MC38 mice treated with 5-FU/PD-L1 ± treracilil combination therapy was measured (n=10-15/treatment group). The x-axis is days of treatment measured in days and the y-axis is percent survival. Figure 5A shows a line graph illustrating tumor growth. Tumor growth was measured in CT26 mice treated with oxaliplatin/PD-L1±traciclib combination therapy (n=10-15/treatment group). The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 5B shows a line graph of survival curves. The overall survival of CT26 mice treated with oxaliplatin/PD-L1±traciclib combination therapy was measured (n=10-15/treatment group). The x-axis is days of treatment measured in days and the y-axis is percent survival. Figure 6 shows the proportion of T _reg at days 5 and 9 of total tumors treated with vehicle, oxaliplatin/PD-L1 or treracilil/oxaliplatin/PD-L1. The x-axis is the results at days 5 and 9 and the y-axis is the % Treg of CD4+ T cells in the tumor measured as a percentage. Figure 7 shows the ratio of CD8 ⁺ T cells to T _regs in tumors (% CD8+ T cells/% T _reg ) (n=5-8 tumors analyzed/treatment group and time point). The x-axis is the results at day 5 and 9 and the y-axis is the ratio of CD8 ⁺ T cells to T _regs measured as a percentage in the tumor. Figure 8 shows the proportion of activated (% CD69+) cells among CD8 ⁺ T cells. *P<0.05. The x-axis shows the treatment groups (vehicle, oxaliplatin/PD-L1 and triaciclib/oxaliplatin/PD-L1). Percentage of activated (% CD69 ⁺ ) cells among CD8 ⁺ T cells on the y axis. Figure 9 shows tumor growth curves of MC38 treated with CDK4/6 inhibitors or PD-1 antibodies (alone or in combination). MC38 murine cancer cells were injected subcutaneously into C57BL/6 mice. Beginning on day 3, use triaciclib (100 mg/kg) intermittently (3 days on, 4 days off) with or without PD-1 antibody (200 µg/mouse, 3 times a week) as directed ) to treat mice (MC38). Tumor volume was monitored every 2-3 days. Each graph shows representative results of two independent experiments. (n=8) (*p<0.001). The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 10A is a bar graph illustrating the quantification of IL-2 cytokine production by MC38 tumor infiltrating T lymphocytes. At the end of treatment (day 17), mice were sacrificed and TILs were isolated from tumors for interleukin analysis of IL-2 in CD4+ T cells (*p<0.001). x-axis is treatment group and y-axis is percentage of IL-2 in CD4 ⁺ cells. Figure 10B is a bar graph illustrating the quantification of IFNy cytokine production by MC38 tumor infiltrating T lymphocytes. At the end of treatment (day 17), mice were sacrificed and TILs were isolated from tumors for interleukin analysis of IFNγ in CD8 ⁺ T cells (*p<0.001). x axis is treated group and y axis is percentage of IFNγ in CD8 ⁺ cells. Figure 11A shows tumors of Balb/C mice implanted with MMTV-PyMT murine tumor cells and then treated with CDK4/6 inhibitors and/or anti-PD-1 inhibitors and/or anti-TIGIT inhibitors (alone or in combination) Growth curve. MMTV-PyMT murine mammary tumor cells were injected subcutaneously into Balb/C mice. From the 7th day after tumor cell administration, use treracilil (100 mg/kg, once a week), anti-PD-1 inhibitors (5 mg/kg, twice a week) and/or anti-TIGIT as directed Mice were treated with inhibitors (10 mg/kg, twice a week) (alone or in combination). The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 1 IB shows tumors of Balb/C mice implanted with MMTV-PyMT murine tumor cells and then treated with CDK4/6 inhibitors and/or anti-PD-1 inhibitors and/or anti-TIGIT inhibitors (alone or in combination) Growth curve. MMTV-PyMT murine mammary tumor cells were injected subcutaneously into Balb/C mice. From the 7th day after tumor cell administration, use treracilil (100 mg/kg, once a week), anti-PD-1 inhibitors (5 mg/kg, twice a week) and/or anti-TIGIT as directed Mice were treated with inhibitors (10 mg/kg, twice a week) (alone or in combination). The x-axis is days of treatment measured in days and the y-axis is tumor growth (fold change). Figure 11C shows the overall survival of Balb/C mice implanted with MMTV-PyMT murine tumor cells and then treated with treracilil and/or anti-PD-1 inhibitors and/or anti-TIGIT inhibitors (alone or in combination) . MMTV-PyMT murine mammary tumor cells were injected subcutaneously into Balb/C mice. From the 7th day after tumor cell administration, CDK4/6 inhibitor (100 mg/kg treracilil, once a week) and anti-PD-1 inhibitor (5 mg/kg, twice a week) were used as directed and/or anti-TIGIT inhibitor (10 mg/kg, twice a week) (alone or in combination) to treat mice. The x-axis is the number of days of treatment measured in days and the y-axis is the probability of survival measured in percent. Figure 1 ID shows tumor growth curves of Balb/C mice (negative control) implanted with MMTV-PyMT murine tumor cells and then treated with citrate buffer. MMTV-PyMT murine mammary tumor cells were injected subcutaneously into Balb/C mice (n=8). Beginning on day 7 after tumor cell administration, mice were treated with citrate buffer as indicated (negative control). The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . FIG. 11E shows tumor growth curves of Balb/C mice implanted with MMTV-PyMT murine tumor cells and then treated with treracilib alone. MMTV-PyMT murine mammary tumor cells were injected subcutaneously into Balb/C mice (n=8). Beginning on day 7 after tumor cell administration, mice were treated with treracilil (100 mg/kg, once a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 1 IF shows tumor growth curves of Balb/C mice implanted with MMTV-PyMT murine tumor cells and then treated with anti-TIGIT inhibitors only. MMTV-PyMT murine mammary tumor cells were injected subcutaneously into Balb/C mice (n=8). From day 7 after tumor cell administration, mice were treated with anti-TIGIT inhibitor (10 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 11G shows tumor growth curves of Balb/C mice implanted with MMTV-PyMT murine tumor cells and then treated with anti-PD-1 inhibitors only. MMTV-PyMT murine mammary tumor cells were injected subcutaneously into Balb/C mice (n=8). From day 7 after tumor cell administration, mice were treated with an anti-PD-1 inhibitor (5 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 11H shows tumor growth curves of Balb/C mice implanted with MMTV-PyMT murine tumor cells and then treated with a combination of anti-PD-1 inhibitor and anti-TIGIT inhibitor. MMTV-PyMT murine mammary tumor cells were injected subcutaneously into Balb/C mice (n=8). From the 7th day after tumor cell administration, anti-PD-1 inhibitor (5 mg/kg, twice a week) and anti-TIGIT inhibitor (10 mg/kg, twice a week) were used to treat small children as directed. mouse. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . FIG. 11I shows tumor growth curves of Balb/C mice implanted with MMTV-PyMT murine tumor cells and then treated with a combination of treracilil and an anti-PD-1 inhibitor. MMTV-PyMT murine mammary tumor cells were injected subcutaneously into Balb/C mice (n=8). From the 7th day after tumor cell administration, mice were treated with treracilil (100 mg/kg, once a week) and anti-PD-1 inhibitor (5 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . FIG. 11J shows tumor growth curves of Balb/C mice implanted with MMTV-PyMT murine tumor cells and then treated with a combination of treracilil and an anti-TIGIT inhibitor. MMTV-PyMT murine mammary tumor cells were injected subcutaneously into Balb/C mice (n=8). From the 7th day after tumor cell administration, mice were treated with treracilil (100 mg/kg, once a week) and anti-TIGIT inhibitor (10 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . FIG. 11K shows tumor growth curves of Balb/C mice implanted with MMTV-PyMT murine tumor cells and then treated with a combination of treracilil, anti-PD1 inhibitor and anti-TIGIT inhibitor. MMTV-PyMT murine mammary tumor cells were injected subcutaneously into Balb/C mice (n=8). From the 7th day after tumor cell administration, use treracicil (100 mg/kg, once a week), anti-TIGIT inhibitor (10 mg/kg, twice a week) and anti-PD-1 inhibitor as directed (5 mg/kg, 2 times a week) to treat mice. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 12A shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with CDK4/6 inhibitors and/or anti-PD1 inhibitors and/or anti-TIGIT inhibitors (alone or in combination). CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 7th day after the administration of tumor cells, use treracilil (100 mg/kg, once a week) as indicated, with or without anti-PD-1 inhibitors (5 mg/kg, 2 times a week) and Mice were treated with or without anti-TIGIT inhibitors (10 mg/kg, twice a week). The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 12B shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with CDK4/6 inhibitors and/or anti-PDl inhibitors and/or anti-TIGIT inhibitors (alone or in combination). CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 7th day after tumor cell administration, use treracilil (100 mg/kg, once a week) as indicated, with or without anti-PD-1 inhibitors (5 mg/kg, twice a week) and use Mice were treated with or without anti-TIGIT inhibitor (10 mg/kg, twice a week). The x-axis is days of treatment measured in days and the y-axis is tumor growth (fold change). Figure 12C shows the overall survival of Balb/C mice implanted with CT26 murine tumor cells and then treated with treracilil and/or anti-PD1 inhibitor and/or anti-TIGIT inhibitor (alone or in combination). CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 7th day after tumor cell administration, use or not use triaciclib (100 mg/kg, once a week), use or not use anti-PD-1 inhibitors (5 mg/kg, once a week) as indicated times) and mice were treated with or without anti-TIGIT inhibitors (10 mg/kg, twice a week). Dosing was terminated on day 63. The x-axis is the number of days of treatment measured in days and the y-axis is the probability of survival measured in percent. Figure 12D shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with citrate buffer (negative control). CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). Beginning on day 7 after tumor cell administration, mice were treated with citrate buffer as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 12E shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with treracilib alone. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). Beginning on day 7 after tumor cell administration, mice were treated with treracilil (100 mg/kg, once a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 12F shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with anti-TIGIT inhibitors only. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From day 7 after tumor cell administration, mice were treated with anti-TIGIT inhibitor (10 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 12G shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with anti-PD-1 inhibitors only. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From day 7 after tumor cell administration, mice were treated with an anti-PD-1 inhibitor (5 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 12H shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of anti-PD-1 inhibitor and anti-TIGIT inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From day 7 after tumor administration, mice were treated with anti-PD-1 inhibitor (5 mg/kg, twice a week) and anti-TIGIT inhibitor (10 mg/kg, twice a week) as indicated . The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 12I shows the tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilil and an anti-PD-1 inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 7th day after tumor cell administration, mice were treated with treracilil (100 mg/kg, once a week) and anti-PD-1 inhibitor (5 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 12J shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilil and an anti-TIGIT inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 7th day after tumor cell administration, mice were treated with treracilil (100 mg/kg, once a week) and anti-TIGIT inhibitor (10 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 12K shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilib, anti-PD-1 inhibitor and anti-TIGIT inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 7th day after tumor cell administration, use treracicil (100 mg/kg, once a week), anti-TIGIT inhibitor (10 mg/kg, twice a week) and anti-PD-1 inhibitor as directed (5 mg/kg, 2 times a week) to treat mice. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . FIG. 13A shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with treracilib and/or anti-PD-1 inhibitors and/or anti-TIGIT inhibitors (alone or in combination). CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 10th day after the administration of tumor cells, use treracilil (100 mg/kg, once a week) as indicated, with or without anti-PD-1 inhibitors (5 mg/kg, 2 times a week) and Mice were treated with or without anti-TIGIT inhibitors (10 mg/kg, twice a week). The x-axis is days of treatment measured in days and the y-axis is tumor growth (fold change). Figure 13B shows the overall survival of Balb/C mice implanted with CT26 murine tumor cells and then treated with treracilil and/or anti-PD-1 inhibitors and/or anti-TIGIT inhibitors (alone or in combination). CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 10th day after the administration of tumor cells, use or not use triaciclib (100 mg/kg, once a week), use or not use anti-PD-1 inhibitors (5 mg/kg, once a week) as indicated times) and mice were treated with or without anti-TIGIT inhibitors (10 mg/kg, twice a week). Dosing was terminated on day 63. The x-axis is the number of days of treatment measured in days and the y-axis is the probability of survival measured in percent. Figure 13C shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with citrate buffer (negative control). CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). Beginning on day 10 after tumor cell administration, mice were treated with citrate buffer as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 13D shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with treracilib alone. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). Beginning on day 10 after tumor cell administration, mice were treated with treracicil (100 mg/kg, once a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 13E shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with anti-TIGIT inhibitors only. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From day 10 after tumor cell administration, mice were treated with anti-TIGIT inhibitor (10 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 13F shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilil and an anti-TIGIT inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). Starting from day 10 after tumor cell administration, mice were treated with treracilil (100 mg/kg, once a week) and anti-TIGIT inhibitor (10 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 13G shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilil and an anti-PD-1 inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From day 10 after tumor cell administration, mice were treated with treracilil (100 mg/kg, once a week) and anti-PD-1 inhibitor (5 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 13H shows the tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilib, anti-PD-1 inhibitor and anti-TIGIT inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 10th day after tumor cell administration, use treracilil (100 mg/kg, once a week), anti-TIGIT inhibitor (10 mg/kg, twice a week) and anti-PD-1 inhibitor as directed (5 mg/kg, 2 times a week) to treat mice. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 14A shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with anti-TIGIT inhibitors. Beginning on day 7 or 10, mice (n=8) were treated with anti-TIGIT inhibitors (10 mg/kg, twice a week) as indicated. The x-axis is post-treatment days measured in days and the y-axis is tumor volume measured in mm ³ . Figure 14B shows the tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilil and an anti-TIGIT inhibitor. Beginning on day 7 or 10, mice were treated (n=8 ). The x-axis is post-treatment days measured in days and the y-axis is tumor volume measured in mm ³ . Figure 14C shows the tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilil, anti-PD-1 inhibitor and anti-TIGIT inhibitor. From the 7th or 10th day, according to the instructions, use treracilil (100 mg/kg, once a week), anti-TIGIT inhibitor (10 mg/kg, twice a week) and anti-PD-1 inhibitor ( 5 mg/kg, twice a week) to treat mice (n=8). The x-axis is post-treatment days measured in days and the y-axis is tumor volume measured in mm ³ . Figure 15A shows Balb/C cells implanted with CT26 murine tumor cells and then treated with treracilil and/or anti-PD-1 inhibitors and/or anti-LAG3 inhibitors and/or anti-TIM3 inhibitors (alone or in combination) Tumor growth curves in mice. CT26 murine colon cancer cells were injected subcutaneously into Balb/c mice (n=8). From day 7 after tumor cell administration, CDK4/6 inhibitors (100 mg/kg treracilil, once a week), with or without anti-PD-1 inhibitors (5 mg/kg, once a week) were used as indicated. Mice were treated with or without anti-LAG3 inhibitor (10 mg/kg, twice a week) and with or without anti-TIM3 inhibitor (5 mg/kg, twice a week). The x-axis is days of treatment measured in days and the y-axis is tumor growth (fold change). Figure 15B shows the tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with treracilil and/or anti-PD-1 inhibitors and/or anti-LAG3 inhibitors (alone or in combination). CT26 murine colon cancer cells were injected subcutaneously into Balb/c mice and then randomized into treatment groups (n=5-8) after reaching a mean tumor volume of 40-80 mm ³ . CDK4/6 inhibitor (100 mg/kg triaciclib once weekly), with or without anti-PD-1 inhibitor (5 mg/kg twice weekly) with or without anti-LAG3 inhibitor (10 mg/kg, 2 times a week) to treat mice. The x-axis is days of treatment measured in days and the y-axis is tumor growth (fold change). Figure 15C shows the tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with treracilil and/or anti-PD-1 inhibitor and/or anti-TIM3 inhibitor (alone or in combination). CT26 murine colon cancer cells were injected subcutaneously into Balb/c mice and then randomized into treatment groups (n=5-8) after reaching a mean tumor volume of 40-80 mm ³ . CDK4/6 inhibitor (100 mg/kg treracilil once weekly), with or without anti-PD-1 inhibitor (5 mg/kg twice weekly) with or without anti-TIM3 inhibitor (10 mg/kg, 2 times a week) to treat mice. The x-axis is days of treatment measured in days and the y-axis is tumor growth (fold change). Figure 15D shows the overall survival of Balb/C mice implanted with CT26 murine tumor cells and then treated with treracilil and/or anti-PD-1 inhibitors and/or anti-LAG3 inhibitors (alone or in combination). CT26 murine colon cancer cells were injected subcutaneously into Balb/c mice (n=8). From the 7th day after tumor cell administration, CDK4/6 inhibitors (100 mg/kg treracilil, once a week), with or without anti-PD-1 inhibitors (5 mg/kg, 2 days a week) were administered. times) with or without anti-LAG3 inhibitors (10 mg/kg, twice a week) to treat mice. The x-axis is the number of days of treatment measured in days and the y-axis is the probability of survival measured in percent. Figure 15E shows the overall survival of Balb/C mice implanted with CT26 murine tumor cells and then treated with treracilil and/or anti-PD-1 inhibitor and/or anti-TIM3 inhibitor (alone or in combination). CT26 murine colon cancer cells were injected subcutaneously into Balb/c mice (n=8). From the 7th day after tumor cell administration, CDK4/6 inhibitors (100 mg/kg treracilil, once a week), with or without anti-PD-1 inhibitors (5 mg/kg, 2 days a week) were administered. times) with or without anti-TIM3 inhibitors (10 mg/kg, twice a week) to treat mice. The x-axis is the number of days of treatment measured in days and the y-axis is the probability of survival measured in percent. Figure 15F shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with citrate buffer (negative control). CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=4). Beginning on day 7 after tumor cell administration, mice were treated with citrate buffer as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 15G shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with treracilib alone. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). Beginning on day 7 after tumor cell administration, mice were treated with a CDK4/6 inhibitor (100 mg/kg treraciclib once a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 15H shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with anti-PD-1 inhibitors only. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From day 7 after tumor cell administration, mice were treated with an anti-PD-1 inhibitor (5 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 15I shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with anti-LAG3 inhibitors only. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From day 7 after tumor cell administration, mice were treated with an anti-LAG3 inhibitor (10 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 15J shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with anti-TIM3 inhibitors only. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From day 7 after tumor cell administration, mice were treated with an anti-TIM3 inhibitor (5 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 15K shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of anti-PD1 inhibitor and anti-LAG3 inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From day 7 after tumor administration, mice were treated with anti-PD1 inhibitor (5 mg/kg, twice a week) and anti-LAG3 inhibitor (10 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 15L shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of anti-PD-1 inhibitor and anti-TIM3 inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From day 7 after tumor administration, mice were treated with anti-PD-1 inhibitor (5 mg/kg, twice a week) and anti-TIM3 inhibitor (5 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 15M shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilil and an anti-PD-1 inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 7th day after tumor cell administration, mice were treated with treracilil (100 mg/kg, once a week) and anti-PD-1 inhibitor (5 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 15N shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilil and an anti-LAG3 inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). Starting from day 7 after tumor cell administration, mice were treated with treaciclib (100 mg/kg, once a week) and anti-LAG3 inhibitor (10 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 15O shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilil and an anti-TIM3 inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 7th day after tumor cell administration, mice were treated with treracilil (100 mg/kg, once a week) and an anti-TIM3 inhibitor (5 mg/kg, twice a week) as indicated. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 15P shows the tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilil, anti-PD-1 inhibitor and anti-LAG3 inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 7th day after tumor cell administration, use treracilil (100 mg/kg, once a week), anti-LAG3 inhibitors (10 mg/kg, twice a week) and anti-PD-1 inhibitors as directed (5 mg/kg, 2 times a week) to treat mice. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 15Q shows the tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with a combination of treracilil, anti-PD-1 inhibitor and anti-TIM3 inhibitor. CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice (n=8). From the 7th day after tumor cell administration, use treracilil (100 mg/kg, once a week), anti-TIM3 inhibitors (5 mg/kg, twice a week) and anti-PD-1 inhibitors as directed (5 mg/kg, 2 times a week) to treat mice. The x-axis is treatment days measured in days and the y-axis is tumor volume measured in ^mm3 . Figure 16A shows Balb/C implanted with CT26 colorectal cancer (CRC) murine tumor cells and then treated with CDK4/6 inhibitors and/or anti-PD-1 inhibitors and/or anti-TIGIT inhibitors (alone or in combination) Tumor growth curves in mice. CT-26 murine colorectal cancer cells were injected subcutaneously into Balb/c mice. From the 10th day after tumor cell implantation, use treracilil (100 mg/kg, once a week) as indicated, with or without anti-PD-1 inhibitors (5 mg/kg, twice a week) and use Mice were treated with or without anti-TIGIT inhibitor (10 mg/kg, twice a week). The x-axis is post-treatment days measured in days and the y-axis is tumor volume measured in mm ³ . Figure 16B shows Balb/C mice implanted with CT26 colorectal cancer (CRC) murine tumor cells and then treated with treracilil and/or anti-PD-1 inhibitors and/or anti-TIGIT inhibitors (alone or in combination) the overall survival period. CT-26 murine colorectal cancer cells were injected subcutaneously into Balb/C mice. From the 10th day after tumor cell administration, use treracilil (100 mg/kg, once a week), anti-PD-1 inhibitors (5 mg/kg, twice a week) and/or anti-TIGIT as directed Mice were treated with inhibitors (10 mg/kg, twice a week) (alone or in combination). The x-axis is the number of days of treatment measured in days and the y-axis is the probability of survival measured in percent. Figure 17A shows tumors of C57BL/6 mice implanted with AT3-OVA murine tumor cells and then treated with CDK4/6 inhibitors and/or anti-PD-1 inhibitors and/or anti-TIGIT inhibitors (alone or in combination) Growth curve. AT3-OVA murine breast cancer cells were injected subcutaneously into C57BL/6 mice. From the 10th day after tumor cell implantation, CDK4/6 inhibitors (100 mg/kg treracilil, once a week), with or without anti-PD-1 inhibitors (5 mg/kg, once a week) were used as indicated. Twice a week) with or without anti-TIGIT inhibitors (10 mg/kg, twice a week) to treat mice. The x-axis is post-treatment days measured in days and the y-axis is tumor volume measured in mm ³ . Figure 17B shows Balb/C mice implanted with AT3-OVA breast cancer (BC) murine tumor cells and then treated with treracilil and/or anti-PD-1 inhibitors and/or anti-TIGIT inhibitors (alone or in combination) the overall survival period. AT3-OVA murine breast cancer cells were injected subcutaneously into Balb/C mice. From the 10th day after tumor cell administration, use treracilil (100 mg/kg, once a week), anti-PD-1 inhibitors (5 mg/kg, twice a week) and/or anti-TIGIT as directed Mice were treated with inhibitors (10 mg/kg, twice a week) (alone or in combination). The x-axis is the number of days of treatment measured in days and the y-axis is the probability of survival measured in percent. Figure 18 shows tumor growth curves of Balb/C mice implanted with S2WTP3 murine tumor cells and then treated with CDK4/6 inhibitors and/or anti-PD-1 inhibitors and/or anti-TIGIT inhibitors (alone or in combination) . S2WTP3 murine breast cancer cells were injected subcutaneously into Balb/c mice. From day 7, CDK4/6 inhibitors (100 mg/kg treracilil, once a week), anti-PD-1 inhibitors (5 mg/kg, twice a week) with or without anti-PD-1 inhibitors were used as directed, and Mice were treated with or without anti-TIGIT inhibitors (10 mg/kg, twice a week). The x-axis is post-treatment days measured in days and the y-axis is tumor volume measured in mm ³ . Figure 19 shows a visual representation of the effects of adenosinergic molecules on tumors and surrounding stroma. FIG. 20A shows tumor growth curves of Balb/C mice implanted with CT26 murine tumor cells and then treated with treracilil and/or anti-PD-1 inhibitors and/or anti-CD73 inhibitors (alone or in combination). CT-26 murine colon carcinoma cells were injected subcutaneously into Balb/c mice. From the 7th day after tumor cell implantation, use treracilil (100 mg/kg, once a week) as indicated, with or without anti-PD-1 inhibitors (5 mg/kg, twice a week) and use Mice were treated with or without anti-CD73 inhibitors (5 mg/kg, twice a week). Continue treatment for 6 weeks. The x-axis is post-treatment days measured in days and the y-axis is tumor volume measured in mm ³ . Figure 20B shows the overall survival of Balb/C mice implanted with CT26 murine tumor cells and then treated with CDK4/6 inhibitors and/or anti-PD-1 inhibitors and/or anti-TIGIT inhibitors (alone or in combination) . CT-26 murine colon cancer cells were injected subcutaneously into Balb/c mice (n=8/group). From the 7th day after tumor cell implantation, CDK4/6 inhibitor (100 mg/kg treracilil, once a week) was used or not, anti-PD-1 inhibitor (5 mg/kg/week) was used or not as indicated. kg, twice a week) with or without anti-CD73 inhibitors (5 mg/kg, twice a week) to treat mice. Continue treatment for 6 weeks. The x-axis is the number of days of treatment measured in days and the y-axis is the probability of survival measured in percent.

Claims (96)

Use of a cyclin-dependent kinase 4/6 (CDK4/6) inhibitor for the manufacture of a medicament for treating a human patient with cancer, wherein the cancer is an advanced or metastatic solid cancer, and wherein the The treatment comprises a. administering to the patient an effective amount of the CDK4/6 inhibitor, wherein the CDK4/6 inhibitor has the following structure: (trilaciclib), or a pharmaceutically acceptable salt thereof; b. Administering an effective amount of programmed cell death protein-1 (PD-1) inhibitor or programmed death ligand-1 to the patient (PD-L1) inhibitors; and, c. administering to the patient an effective amount of another immune checkpoint inhibitor selected from the group consisting of T cell immune receptor inhibitors with immunoglobulin and ITIM domains (TIGIT) , T cell immunoglobulin and mucin domain 3 (TIM-3) inhibitors, lymphocyte activation gene-3 (LAG-3) inhibitors or cluster of differentiation 73 (CD73) inhibitors. The use according to claim 1, wherein the method further comprises administering an effective amount of a chemotherapeutic agent to the patient. The use according to claim 1 or 2, wherein the solid cancer is selected from small cell lung cancer, non-small cell lung cancer, triple negative breast cancer, colorectal cancer, urothelial cancer, cervical cancer, esophageal cancer or squamous cell carcinoma of the head and neck . The use according to claims 1 to 3, wherein the treatment is administered to the patient in a first-line setting. The use according to claims 1 to 3, wherein the treatment is administered to the patient in a second line setting. The use according to claim 5, wherein the patient has previously received a PD-1 or PD-L1 inhibitor and has experienced disease progression. The use according to claims 1 to 6, wherein an effective amount of a PD-1 inhibitor is administered to the patient. The use of claim 7, wherein the PD-1 inhibitor is selected from nivolumab, pembrolizumab, cemiplimab, dotalimab (dostarlimab), pidilizumab, AMP-224, AMP-514, sintilimab, sasanlimab, spartalizumab, Rifeli Retifanlimab, tislelizumab, toripalimab, camrelizumab, CS1003, zimberelimab, or JTX-4014 . The use according to claims 1 to 8, wherein an effective amount of a PD-L1 inhibitor is administered to the patient. The use of claim 9, wherein the PD-L1 inhibitor is selected from atezolizumab, durvalumab, avelumab, envolimumab ( envafolimab), BMS-936559, BMS-986189, lodapolimab, cosibelimab, sugemalimab, adebrelimab, CBT-502, AUNP12 , CA-170 or BGB-A333. The use according to claims 1 to 10, wherein the other immune checkpoint inhibitor administered is a LAG-3 inhibitor. The use of claim 11, wherein the LAG-3 inhibitor is selected from the group consisting of relatlimab, GSK2831781, eftilagimod alpha, leramilimab, French Favezelimab, fianlimab, TSR-033, BI754111, Sym022, LBL-007, IBI110, IBI323, INCAGN02385, AVA021, MGD013, RO7247669, EMB-02, AVA-0017, XmAb841 , tebotelimab (tebotelimab), FS118, CB213 or SNA-03. The use according to claims 1 to 10, wherein the other immune checkpoint inhibitor administered is a TIM-3 inhibitor. The use of claim 13, wherein the TIM-3 inhibitor is selected from cobolimab, RG7769, MAS825, sabatolimab, Sym023, INCAGN2390, LY3321367, BMS-986258, SHR -1702, AZD7789, TQB2618, NB002, BGBA425 or RO7121661. The use according to claims 1 to 10, wherein the other immune checkpoint inhibitor administered is a TIGIT inhibitor. The use of claim 15, wherein the TIGIT inhibitor is selected from BAT6005, Vibostolimab, Etigilimab, Tiragolumab, Ospelimab (ociperlimab), BMS-986207, COM902, M6223, domvanalimab, AZD2936, JS006, IBI139, ASP-8374, BAT6021, TAB006, EOS884448, SEA-TGT, mAb-7, SHR-1708, GS02 , RXI-804, NB6253, ENUM009, CASC-674, AJUD008, AGEN1777, HLX301, HLX53, M6223 or HB0036. The use according to claims 1 to 10, wherein the other immune checkpoint inhibitor administered is a CD73 inhibitor. The use of claim 17, wherein the CD73 inhibitor is selected from HLX23, LY3475070, IPH5301, AK119, PT199, mupadolimab, Sym024, oleclumab, IBI325, ORIC-533 , JAB-BX102, TJ004309, AB680, NZV930, BMS-986179, INCA00186 or dalutrafusp alfa. For the purposes of claims 1 to 18, wherein the triracilli is administered once a week. The use according to claims 1 to 19, wherein the PD-1 inhibitor or PD-L1 inhibitor and the other immune checkpoint inhibitors are administered once every two weeks. The use according to claims 1 to 19, wherein the PD-1 inhibitor or PD-L1 inhibitor and the other immune checkpoint inhibitors are administered once every three weeks. The use according to claims 1 to 19, wherein the PD-1 inhibitor or PD-L1 inhibitor and the other immune checkpoint inhibitors are administered once every four weeks. The use according to claims 1 to 19, wherein the PD-1 inhibitor or PD-L1 inhibitor and the other immune checkpoint inhibitors are administered once every six weeks. The use of claims 1 to 19, wherein the PD-1 inhibitor or PD-L1 inhibitor is administered once for the duration of the first cycle and the other immune checkpoint inhibitor is administered for the duration of the second cycle Introject once; wherein the duration of the first period is selected from two weeks, three weeks, four weeks or six weeks; wherein the duration of the second period is selected from two weeks, three weeks, four weeks or six weeks; and Wherein the duration of the first period is different from the duration of the second period. The use according to claim 1, wherein a PD-1 inhibitor is administered and a LAG-3 inhibitor is administered, and wherein the PD-1 inhibitor is Nivolumab and the LAG-3 inhibitor is Rilarizumab anti. The use according to claims 2 to 25, wherein treraxilib is administered about 8 hours or less before administering the chemotherapeutic agent. The use according to claim 26, wherein treracicil is administered about 4 hours or less before administering the chemotherapeutic agent. The purposes of claims 26 to 27, wherein the chemotherapeutic agent is selected from platinum drugs, taxanes, topoisomerase inhibitors, alkylating agents, cytotoxic antibiotics, antimetabolites or vinca organisms alkali. As the purposes of claim 26 to 27, wherein the chemotherapy agent is selected from carboplatin (carboplatin), cisplatin (cisplatin), oxaliplatin (oxaliplatin), paclitaxel (paclitaxel), docetaxel (docetaxel) , Paclitaxel nanoparticle formulation stabilized by albumin (nab-paclitaxel), topotecan, campothecin, irinotecan, belotecan, Doxorubicin, daunorubicin, epirubicin, idarubicin, etoposide, teniposide, cyclophosphamide (cyclophosphamide), vinblastine, gemcitabine, 5-fluorouracil, eribulin, pemetrexed, mitomycin, goxa Sacituzumab govitecan, valrubicin, vinorelbine tartrate, trabectedin, temozolomide, melphalan, dacarbazine ( dacarbazine, methotrexate, mitroxantrone, bleomycin, irinotecan, cabazitaxel, bortezomib, vincristine ), vindesine, diaziquone, mechlorethamine, mitomycin C, fludarabine, cytosine arabinoside , ixabepilone, enfortumab vedotin, ifosfamide, capecitabine, or any pharmaceutically acceptable salt thereof; or its combination. The use according to claims 1 to 29, wherein the solid cancer expresses PD-L1. Use of a CDK4/6 inhibitor for the manufacture of a medicament for the treatment of a human patient with cancer, wherein the cancer is an advanced or metastatic solid cancer, and wherein the treatment comprises a. Administering an effective amount of the CDK4/6 inhibitor triraciclib or a pharmaceutically acceptable salt thereof to the patient; b. Administering an effective amount of a PD-1 inhibitor or PD-L1 inhibitor to the patient; and, c. Administering an effective amount of a TIM-3 inhibitor to the patient. The use according to claim 31, wherein the method further comprises administering an effective amount of a chemotherapeutic agent to the patient. The use of claim 31 or 32, wherein the solid cancer is selected from colorectal cancer, small cell lung cancer, non-small cell lung cancer, triple negative breast cancer, urothelial cancer, cervical cancer, esophageal cancer or squamous cell carcinoma of the head and neck . The use according to claims 31 to 33, wherein the treatment is administered to the patient in a first-line setting. The use according to claims 31 to 33, wherein the treatment is administered to the patient in a second line setting. The use according to claims 31 to 35, wherein the patient has previously received a PD-1 inhibitor or a PD-L1 inhibitor and has experienced disease progression. The use according to claims 31 to 36, wherein an effective amount of a PD-1 inhibitor is administered to the patient. Such as the use of claim 37, wherein the PD-1 inhibitor is selected from nivolumab, pembrolizumab, simiprizumab, dotalimab, pilizumab, AMP- 224, AMP-514, sintilimab, sasanlimab, sparizumab, revelizumab, tislelizumab, toripalimab, camrelizumab, CS1003, Zimbelizumab, or JTX-4014. The use according to claims 31 to 36, wherein an effective amount of a PD-L1 inhibitor is administered to the patient. Such as the use of claim 39, wherein the PD-L1 inhibitor is selected from atezolizumab, durvalumab, avelumab, envolimumab, BMS-936559, BMS-986189, Luo Daclizumab, coxelimumab, sugemalimumab, adelbelimab, CBT-502, AUNP12, CA-170, or BGB-A333. The use of claim items 31 to 40, wherein the TIM-3 inhibitor is selected from the group consisting of Coblemumab, RG7769, MAS825, Sabatolizumab, Sym023, INCAGN2390, LY3321367, BMS-986258, SHR-1702, AZD7789 , TQB2618, NB002, BGBA425 or RO7121661. As the use of claims 31 to 41, wherein the triracilli is administered once a week. The use according to claims 31 to 42, wherein the PD-1 inhibitor or PD-L1 inhibitor and the TIM-3 inhibitor are administered once every two weeks. The use according to claims 31 to 42, wherein the PD-1 inhibitor or PD-L1 inhibitor and the TIM-3 inhibitor are administered once every three weeks. The use according to claims 31 to 42, wherein the PD-1 inhibitor or PD-L1 inhibitor and the TIM-3 inhibitor are administered once every four weeks. The use according to claims 31 to 42, wherein the PD-1 inhibitor or PD-L1 inhibitor and the TIM-3 inhibitor are administered once every six weeks. The use according to claims 31 to 42, wherein the PD-1 inhibitor or PD-L1 inhibitor is administered once within the duration of the first cycle and the TIM-3 inhibitor is administered within the duration of the second cycle cast once; wherein the duration of the first period is selected from two weeks, three weeks, four weeks or six weeks; wherein the duration of the second period is selected from two weeks, three weeks, four weeks or six weeks; and Wherein the duration of the first period is different from the duration of the second period. The use according to claims 31 to 47, wherein treracicil is administered about 8 hours or less before administering the chemotherapeutic agent. The use according to claim 48, wherein treracicil is administered about 4 hours or less before administering the chemotherapeutic agent. The use according to claims 48 to 49, wherein the chemotherapeutic agent is selected from platinum drugs, taxanes, topoisomerase inhibitors, alkylating agents, cytotoxic antibiotics, antimetabolites or vinca alkaloids. As the purposes of claims 48 to 49, wherein the chemotherapeutic agent is selected from carboplatin, cisplatin, oxaliplatin, paclitaxel, docetaxel, albumin-stabilized paclitaxel nanoparticle formulation (nab - paclitaxel), topotecan, camptothecin, irinotecan, belotecan, doxorubicin, daunorubicin, epirubicin, idarubicin, etoposide, teniposide , cyclophosphamide, vinblastine, gemcitabine, 5-fluorouracil, eribulin, pemetrexed, mitomycin, gosatuzumab, valrubicin, vinorelbine tartrate, trabectin Dine, temozolomide, melphalan, dacarbazine, methotrexate, mitoxantrone, bleomycin, irinotecan, cabazitaxel, bortezomib, vincristine, vindesine, decacquinone , bischloroethylmethylamine, mitomycin C, fludarabine, cytosine arabinoside, ixabepilone, envertumumab-vedotin, ifosfamide and capecita Bin or any pharmaceutically acceptable salt thereof; or a combination thereof. The use according to claims 31 to 51, wherein the solid cancer expresses PD-L1. Use of a CDK4/6 inhibitor for the manufacture of a medicament for the treatment of a human patient with cancer, wherein the cancer is an advanced or metastatic solid cancer, and wherein the treatment comprises a. Administering an effective amount of the CDK4/6 inhibitor treracilil or a pharmaceutically acceptable salt thereof to the patient; b. Administering an effective amount of a PD-1 inhibitor or PD-L1 inhibitor to the patient; and, c. Administering an effective amount of a LAG-3 inhibitor to the patient. The use according to claim 53, wherein the method further comprises administering an effective amount of a chemotherapeutic agent to the patient. The purposes of claims 53 to 54, wherein the solid cancer is selected from colorectal cancer, small cell lung cancer, non-small cell lung cancer, triple negative breast cancer, urothelial cancer, cervical cancer, esophageal cancer or squamous cell carcinoma of the head and neck . The use according to claims 53 to 55, wherein the treatment is administered to the patient in a first-line setting. The use according to claims 53 to 55, wherein the treatment is administered to the patient in a second line setting. The use according to claim 57, wherein the patient has previously received a PD-1 inhibitor or a PD-L1 inhibitor and has experienced disease progression. The use according to claims 53 to 58, wherein an effective amount of a PD-1 inhibitor is administered to the patient. Such as the use of claim 59, wherein the PD-1 inhibitor is selected from nivolumab, pembrolizumab, simiprizumab, dotalimab, pilizumab, AMP- 224, AMP-514, sintilimab, sasanlimab, sparizumab, revelizumab, tislelizumab, toripalimab, camrelizumab, CS1003, Zimbelizumab, or JTX-4014. The use according to claims 53 to 58, wherein an effective amount of a PD-L1 inhibitor is administered to the patient. Such as the use of claim 61, wherein the PD-L1 inhibitor is selected from atezolizumab, durvalumab, avelumab, envolimumab, BMS-936559, BMS-986189, Luo Daclizumab, coxelimumab, sugemalimumab, adelbelimab, CBT-502, AUNP12, CA-170, or BGB-A333. The use of claims 53 to 62, wherein the LAG-3 inhibitor is selected from the group consisting of Rilarimab, GSK2831781, Eferagimod α, Liramumab, Favizlimab, Fuanli Monoclonal antibody, TSR-033, BI754111, Sym022, LBL-007, IBI110, IBI323, INCAGN02385, AVA021, MGD013, RO7247669, EMB-02, AVA-0017, XmAb841, tepolizumab, FS118, CB213, or SNA-03 . The use as claimed in items 53 to 63, wherein the triraciclib is administered once a week. The use according to claims 53 to 63, wherein the PD-1 inhibitor or PD-L1 inhibitor and the LAG-3 inhibitor are administered once every two weeks. The use according to claims 53 to 63, wherein the PD-1 inhibitor or PD-L1 inhibitor and the LAG-3 inhibitor are administered once every three weeks. The use of claims 53 to 63, wherein the PD-1 inhibitor or PD-L1 inhibitor and the LAG-3 inhibitor are administered once every four weeks. The use according to claims 53 to 63, wherein the PD-1 inhibitor or PD-L1 inhibitor and the LAG-3 inhibitor are administered once every six weeks. The use of claims 53 to 63, wherein the PD-1 inhibitor or PD-L1 inhibitor is administered once within the duration of the first cycle and the LAG-3 inhibitor is administered within the duration of the second cycle cast once; wherein the duration of the first period is selected from two weeks, three weeks, four weeks or six weeks; wherein the duration of the second period is selected from two weeks, three weeks, four weeks or six weeks; and Wherein the duration of the first period is different from the duration of the second period. The use according to claims 54 to 69, wherein treracicil is administered about 8 hours or less before administering the chemotherapeutic agent. The use according to claim 70, wherein treracicil is administered about 4 hours or less before administering the chemotherapeutic agent. The use according to claims 70 to 71, wherein the chemotherapeutic agent is selected from platinum drugs, taxanes, topoisomerase inhibitors, alkylating agents, cytotoxic antibiotics, antimetabolites or vinca alkaloids. As the purposes of claims 70 to 71, wherein the chemotherapeutic agent is selected from carboplatin, cisplatin, oxaliplatin, paclitaxel, docetaxel, albumin-stabilized paclitaxel nanoparticle formulation (nab - paclitaxel), topotecan, camptothecin, irinotecan, belotecan, doxorubicin, daunorubicin, epirubicin, idarubicin, etoposide, teniposide , cyclophosphamide, vinblastine, gemcitabine, 5-fluorouracil, eribulin, pemetrexed, mitomycin, gosatuzumab, valrubicin, vinorelbine tartrate, trabectin Dine, temozolomide, melphalan, dacarbazine, methotrexate, mitoxantrone, bleomycin, irinotecan, cabazitaxel, bortezomib, vincristine, vindesine, decacquinone , bischloroethylmethylamine, mitomycin C, fludarabine, cytosine arabinoside, ixabepilone, envertumumab-vedotin, ifosfamide and capecita Bin or any pharmaceutically acceptable salt thereof; or a combination thereof. The use according to claims 53 to 73, wherein the solid cancer PD-L1. Use of a CDK4/6 inhibitor for the manufacture of a medicament for the treatment of a human patient with cancer, wherein the cancer is an advanced or metastatic solid cancer, and wherein the treatment comprises a. Administering an effective amount of the CDK4/6 inhibitor triraciclib or a pharmaceutically acceptable salt thereof to the patient; b. Administering an effective amount of a PD-1 inhibitor or PD-L1 inhibitor to the patient; and, c. Administering an effective amount of a CD73 inhibitor to the patient. The use according to claim 75, wherein the method further comprises administering an effective amount of a chemotherapeutic agent to the patient. The use of claim 75 or 76, wherein the solid cancer is selected from colorectal cancer, small cell lung cancer, non-small cell lung cancer, triple negative breast cancer, urothelial cancer, cervical cancer, esophageal cancer or squamous cell carcinoma of the head and neck . The use according to claims 75 to 77, wherein the treatment is administered to the patient in a first-line setting. The use according to claims 75 to 77, wherein the treatment is administered to the patient in a second line setting. The use according to claim 79, wherein the patient has previously received a PD-1 inhibitor or a PD-L1 inhibitor and has experienced disease progression. The use according to claims 75 to 80, wherein an effective amount of a PD-1 inhibitor is administered to the patient. Such as the use of claim 81, wherein the PD-1 inhibitor is selected from nivolumab, pembrolizumab, simiprizumab, dotalimab, pilizumab, AMP- 224, AMP-514, sintilimab, sasanlimab, sparizumab, revelizumab, tislelizumab, toripalimab, camrelizumab, CS1003, Zimbelizumab, or JTX-4014. The use according to claims 75 to 80, wherein an effective amount of a PD-L1 inhibitor is administered to the patient. Such as the use of claim 83, wherein the PD-L1 inhibitor is selected from atezolizumab, durvalumab, avelumab, envolimumab, BMS-936559, BMS-986189, Luo Daclizumab, coxelimumab, sugemalimumab, adelbelimab, CBT-502, AUNP12, CA-170, or BGB-A333. Such as the use of claims 75 to 84, wherein the CD73 inhibitor is selected from HLX23, LY3475070, IPH5301, AK119, PT199, mupadolimab, Sym024, Olegluzumab, IBI325, ORIC-533, JAB-BX102 , TJ004309, AB680, NZV930, BMS-986179, INCA00186 or Darovup α. As the use of claims 75 to 85, wherein the triracilli is administered once a week. The use according to claims 75 to 86, wherein the PD-1 inhibitor or PD-L1 inhibitor and the CD73 inhibitor are administered once every two weeks. The use according to claims 75 to 86, wherein the PD-1 inhibitor or PD-L1 inhibitor and the CD73 inhibitor are administered once every three weeks. The use according to claims 75 to 86, wherein the PD-1 inhibitor or PD-L1 inhibitor and the CD73 inhibitor are administered once every four weeks. The use according to claims 75 to 86, wherein the PD-1 inhibitor or PD-L1 inhibitor and the CD73 inhibitor are administered once every six weeks. The use of claims 75 to 86, wherein the PD-1 inhibitor or PD-L1 inhibitor is administered once for the duration of the first cycle and the CD73 inhibitor is administered for the duration of the second cycle once; wherein the duration of the first period is selected from two weeks, three weeks, four weeks or six weeks; wherein the duration of the second period is selected from two weeks, three weeks, four weeks or six weeks; and Wherein the duration of the first period is different from the duration of the second period. The use according to claims 76 to 91, wherein treracicil is administered about 8 hours or less before administering the chemotherapeutic agent. The use according to claim 92, wherein treraxilib is administered about 4 hours or less before administering the chemotherapeutic agent. The use of claims 92 to 93, wherein the chemotherapeutic agent is selected from platinum drugs, taxanes, topoisomerase inhibitors, alkylating agents, cytotoxic antibiotics, antimetabolites or vinca alkaloids. As the purposes of claims 92 to 93, wherein the chemotherapeutic agent is selected from carboplatin, cisplatin, oxaliplatin, paclitaxel, docetaxel, albumin-stabilized paclitaxel nanoparticle formulation (nab - paclitaxel), topotecan, camptothecin, irinotecan, belotecan, doxorubicin, daunorubicin, epirubicin, idarubicin, etoposide, teniposide , cyclophosphamide, vinblastine, gemcitabine, 5-fluorouracil, eribulin, pemetrexed, mitomycin, gosatuzumab, valrubicin, vinorelbine tartrate, trabectin Dine, temozolomide, melphalan, dacarbazine, methotrexate, mitoxantrone, bleomycin, irinotecan, cabazitaxel, bortezomib, vincristine, vindesine, decacquinone , bischloroethylmethylamine, mitomycin C, fludarabine, cytosine arabinoside, ixabepilone, envertumumab-vedotin, ifosfamide and capecita Bin or any pharmaceutically acceptable salt thereof; or a combination thereof. The use according to claims 75 to 95, wherein the solid cancer expresses PD-L1.