Patient selection for enhancing antitumor immunity in cancer patients

Abstract

1. A method for extending the progression-free survival or overall survival of a cancer patient, the method comprising the following steps: determining whether the cancer has a surrounding microenvironment that is favorable for immune modulation; determining whether the chemotherapy regimen induces immunogenic cell death; and if both are positive, administering an effective amount of a CDK4 / 6 inhibitor compound selected from I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, prior to administration of the chemotherapy or, optionally, prior to and concurrently with the chemotherapy, wherein the extension of the progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of chemotherapy alone, based either on literature or otherwise published evidence, comparisons in preclinical or clinical trials, or other means recognized by those skilled in the art.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 62 / 863,153, filed June 18, 2019; and U.S. Provisional Application No. 62 / 907,375, filed September 27, 2019, the entire contents of each of which are incorporated herein by reference for all purposes.

[0002] FIELD OF THE INVENTION The present invention relates to the field of cancer treatment and provides methods for selecting patients for beneficial cancer-directed treatment, including administering a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in combination with chemotherapy, based on patient and cancer profiles, as described in detail herein. It has been found that when a CDK4 / 6 inhibitor is administered in combination with chemotherapy to a specific subsection of cancer patients, this selected patient population exhibits a benefit in progression-free survival and / or overall survival. In some embodiments, this result can be achieved without the use of immune checkpoint inhibitors, such as antibodies anti-PD-1, anti-PD-L1, or anti-CTLA4. It has also been found that administering a CDK4 / 6 inhibitor in combination with chemotherapy to various specific subsections of cancer patients provides a myeloprotective effect, protecting immune cells and potentially resulting in higher rates of T and / or B cells compared to untreated patients, and potentially improving patient experience and quality of life, even if it does not result in improved overall survival. [Background technology]

[0003] The tumor microenvironment (TME) is composed of various cellular and noncellular components within and surrounding a tumor. The TME is known to play an important role in tumor progression. It shapes tumor development (whether the tumor regresses, acquires resistance, evades the immune system, and / or metastasizes), thereby influencing patient outcomes. Chen et al., New horizons in tumor microenvironment biology: challenges and opportunities. BMC Med. 2015 Mar 5;13:45. doi: 10.1186 / s12916-015-0278-7. A correlation has been found between levels of tumor-infiltrating immune cells, a key component of the TME, and patient prognosis. Studies in colorectal cancer have shown that high levels of tumor-infiltrating CD3+ immune cells are associated with improved disease-free survival. Galon et al., Type, density, and location of immune cells within human colorectal tumors predict clinical outcome. Science. 2006 Sep 29;313(5795):1960-4.

[0004] It has recently been recognized that the effects of chemotherapy are highly complex, affecting not only the tumor but also the patient's immune cells, which normally play a major role in protecting the body from diseased cells. Therefore, chemotherapy protocols must consider not only the effects on the tumor but also on the tumor microenvironment.

[0005] It has also been found that certain, but not all, chemotherapeutic agents can induce a pathway in tumor cells called "immunogenic cell death (ICD)" (see, generally, Locy, H., et al., Immunomodulation of the Tumor Microenvironment: Turn Foe Into Friend, Frontiers in Immunology, 2018;9:2090). ICD is a type of regulated cell death that triggers the release of tumor-associated antigens and triggers an antitumor immune response (ibid.). ICD involves the release of damage-associated molecular pathways (DAMPs), which alert the host immune system to cell damage. Six DAMPs promote cell death: calreticulin (CRT), high mobility group box 1 (HMGB1), extracellular ATP, type I interferon, cancer cell-derived nucleic acids, and ANXA1. These DAMPs determine the strength and durability of the ICD antitumor response. See also Wang, et al, Immunogenic effects of chemotherapy-induced tumor cell death, Genes & Diseases (2018) 5, 194-203.

[0006] Chemotherapeutic agents may also induce immunogenic effects by disrupting strategies used by tumors to evade immune responses. See, e.g., Emens et al., The Interplay of Immunotherapy and Chemotherapy: Harnessing Potential Synergies. Cancer Immunol Res; 3(5) May 2015. For example, chemotherapy can modulate salient immunobiological features of tumors in drug-, dose-, and schedule-dependent manners, and salient chemotherapeutic agents can modulate the intrinsic immunogenicity of tumor cells through various mechanisms (see, e.g., Chen G, Emens LA. Chemoimmunotherapy: reengineering tumor immunity. Cancer Immunol Immunother 2013;62:203-16). Chemotherapy can also promote tumor antigen presentation by upregulating the expression of tumor antigens themselves or MHC class I molecules to which the antigens bind. Alternatively, chemotherapy can upregulate costimulatory molecules (B7-1) or downregulate co-inhibitory molecules (PD-L1 / B7-H1 or B7-H4) expressed on the surface of tumor cells, enhancing the potency of effector T cell activity. Chemotherapy also sensitizes tumor cells to T cell-mediated lysis via Fas-, perforin-, and granzyme B-dependent mechanisms.

[0007] Furthermore, recent insights into the fundamental mechanisms of tumor-immune system interactions have enabled the development of tumor classification systems based on the characteristics of individual tumor microenvironments, in relation to immune effector cell populations and the presence or absence of certain immunogenic biomarkers and signals. In 2009, Camus et al. reported a study on colorectal cancer using the categories of hot, altered, and cold. The 2-year recurrence rates for these tumors were 10%, 50%, and 80%. Camus, M., et al., Coordination of intratumoral immune reaction and human colorectal cancer recurrence, Cancer Research 69, 2685-2693 (2009). They further classified altered tumors as excluded or immunosuppressed. They found that in some tumors, T cells were present at the tumor front but failed to infiltrate (i.e., altered excluded), which in itself enabled tumor protection. In other cases, the tumors had a low degree of immune infiltration, suggesting a low degree of leading-edge barrier but an immunosuppressive environment (i.e., altered immunosuppressed). This tumor categorization is now being accepted as a means of predicting progression not only in colorectal cancer but also in other cancer fields.

[0008] Galon and Bruni expanded the classification of tumors into four categories: hot, altered-excluded, altered-immunosuppressed, and cold, to stimulate research and communication. Specifically, the categorization is based on the type, density, and location of immune cells within the tumor site (see Figure 7a). The authors use a scoring system called "Immunoscore," based on the quantification of two lymphocyte populations (CD3 and CD8) in both the tumor center and the tumor's leading edge, to classify tumors according to immune infiltration rather than cancer type. Scores range from I0 (low density, absence of both cell types in both areas) to I4 (high density of immune cell types in both locations). I4 tumors are considered "hot," and I0 tumors are considered "cold." Tumor progression (T stage) and invasion (N stage) were reported to depend on this pre-existing intratumoral adaptive immunity. Researchers are now increasingly focusing on the characteristics, density, immunofunctional orientation, and distribution of immune cells within tumors. See Galon, J., and Bruni, D., “Approaches to treat immune hot, altered, and cold tumors with combination immunotherapies,” Nature Reviews Drug Discovery (18), March 2019, 197-218.

[0009] As reported by Galon, the cardinal features of immunologically hot tumors are (i) extensive infiltration of T cells and cytotoxic T cells, (ii) checkpoint activation or T cell dysfunction. Immunologically altered immune tumors (altered-immunosuppressed immune tumors) are classified by (i) insufficient but not absent infiltration of T cells and cytotoxic T cells, (ii) the presence of soluble inhibitory mediators, (iii) the presence of immunosuppressive cells, and (iv) the presence of T cell checkpoints. Immunologically excluded immune tumors (altered-excluded immune tumors) are characterized by (i) the absence of significant T cell infiltration within the tumor but accumulation of T cells at the tumor periphery, (ii) activation of oncogenic pathways, (iii) epigenetic regulation and reprogramming of the tumor microenvironment, (iv) abnormalities in the tumor vasculature and / or stroma, and (v) hypoxia. Cold tumors are characterized by (i) the absence of T cells within and at the tumor edge, and (ii) unsuccessful T cell priming (i.e., poor, little, or no antigen presentation, low tumor mutational burden, and / or intrinsic insensitivity to T cell cytotoxicity). Cold tumors may also exhibit low PD-L1 expression.

[0010] As shown in Figure 6 herein and reported in their March 2019 Nature Reviews paper on page 204 (see Figure 3 in Galon et al.), Galon et al. present a comprehensive wheel diagram of four categories of tumors, the mechanisms tumor cells use to protect themselves, and the drugs / therapies that can be used to overcome that protection.

[0011] Based on extensive immunogenomic analysis of over 10,000 tumors comprising 33 diverse cancer types, Thorsson et al. identified six immune subtypes that encompass many tumor types. See Thorsson et al., "The Immune Landscape of Cancer," Immunity 48, 812-830, 2018. The six immune subtypes are: C1 - "wound-healing," characterized by a high proliferation rate, high angiogenic gene expression, and a Th2 cell bias toward adaptive immune infiltrates; C2 - "IFN-γ-dominant," characterized by the highest M1 / M2 macrophage polarization, strong CD8 signaling, and high TCR diversity; C3 - "inflammatory," characterized by elevated Th17 and Th1 genes, low-to-moderate proliferation, low aneuploidy, and generalized somatic copy number alterations; C4 - "lymphocyte-depleted," characterized by a prominent macrophage signature with Th1 suppression and a high M2 response; C5 - "immunologically silent," characterized by a low lymphocyte response and a high macrophage response with an M2 predominance; and C6 - "TGF-β-dominant," characterized by a mixed tumor subgroup with high TGF-β and lymphocyte infiltration. Thorsson et al. found that immune subtypes were associated with overall survival (OS) and progression-free survival (PFI), with cancers falling within the C3 classification having the best prognosis, whereas cancers classified as C2 or C1 had less favorable outcomes despite having a substantial immune component, and the more mixed signature subtypes C4 and C6 had the poorest outcomes.

[0012] Ayers et al. analyzed gene expression profiles (GEPs) using RNA from pretreatment baseline tumor samples from PD-1-treated patients and identified immune-related signatures that correlated with clinical activity across nine cancer types. See Ayers et al., “IFN-γ-related mRNA profile predicts clinical response to PD-1 blockade. J Clin Invest. 2017;127(8):2930-2940.” They found that the T cell-inflammatory GEP included IFN-γ-responsive genes associated with antigen presentation, chemokine expression, cytotoxic activity, and adaptive immune tolerance, and that these signatures were necessary but not always sufficient to obtain clinical benefit from the use of checkpoint inhibitors. They identified a subset of six genes (the “IFN-γ signature”) and an additional 18 genes (the “extended immune signature”) whose expression profiles provided predictive value for determining the efficacy of treatment with monoclonal antibodies directed against PD-1 / PD-L1.

[0013] Despite our improved understanding of the effects of chemotherapy on the tumor microenvironment and advances in the field of tumor classification to further our understanding and improve patient outcomes, it is clear that further research and discovery is needed to accurately select patient populations that will benefit from cancer therapy and how they will benefit. The many complex factors involved in driving cancer therapy make this goal challenging and difficult to predict. Summary of the Invention [Problem to be solved by the invention]

[0014] One goal is to be able to select patient populations that may confer progression-free survival benefit and / or overall survival benefit.

[0015] Another goal is to select a patient population in which the therapy may improve the patient experience or quality of life, whether that be a benefit to progression-free survival or an overall benefit, and may provide a myeloprotective effect that protects immune cells. [Means for solving the problem]

[0016] The present invention addresses the problem of patient selection for specific cancer therapy outcomes when patients are administered a cyclin-dependent kinase 4 / 6 inhibitor in combination with chemotherapy.

[0017] It has been found that when a subsection of cancer patients is administered a CDK4 / 6 inhibitor in combination with chemotherapy, this selected patient population exhibits a benefit in progression-free survival and / or overall survival. In some embodiments, this result can be obtained without the use of immune checkpoint inhibitors, such as anti-PD-1, anti-PD-L1, or anti-CTLA4 drugs, e.g., antibodies. For example, if cancer patients have tumors that exhibit certain characteristics as described herein according to the Ayers interferon-γ signature, the Ayers extended immune signature, or the Thorsson et al. six-class immune signature, the patient population is more likely to experience a benefit in progression-free survival or overall survival when administered a CDK4 / 6 inhibitor in combination with chemotherapy. In one embodiment, the tumor is interferon-γ (IFN-γ) dominant according to the Thorsson six-class immune signature, or has a high IFN-γ signature or extended immune signature according to the Ayers IFN-γ signature score or extended immune signature score.

[0018] It has also been found that administering a CDK4 / 6 inhibitor in combination with chemotherapy to a subsection of various specific cancer patients can provide a myeloprotective effect that protects immune cells in select patient populations, resulting in a higher percentage of T cells and / or B cells than without the therapy. In one embodiment, the subsection of various specific cancer patients for whom a myeloprotective effect is provided has non-small cell lung cancer. This patient population includes a patient population with cancer that is particularly non-immunogenic or insensitive to immunomodulation, according to characteristics described in the Background Art or otherwise herein. In one embodiment, the cancer is poorly immunogenic, with relatively low PD-L1 expression (less than about 50%, 40%, or even 30% of normal expression). In another embodiment, the tumor has reduced expression of major histocompatibility complex class I and class II molecules, a known immune evasion mechanism reflecting a poorly immunogenic environment.

[0019] Thus, the present invention provides a means for determining therapy outcomes and, therefore, treatment protocols using appropriate selection of tumor types, chemotherapy types, and anti-cyclin-dependent kinase (CDK) therapy and administration regimen combinations to maximize anti-tumor immunity. This benefit may include enhanced general immune surveillance, as well as reversal of T cell exhaustion, enhanced immune cell activation, including T cells, the formation of immunological memory, and / or reduced immunosuppression. In some embodiments, this result can be achieved without the use of immune checkpoint inhibitors, e.g., anti-PD-1, anti-PD-L1, or anti-CTLA4 agents, e.g., antibodies. Importantly, extending progression-free survival and / or overall survival without the need for checkpoint inhibitor compounds may reduce potential side effects associated with immune checkpoint inhibitor administration, including pneumonia, hyperthyroidism, hypothyroidism, kidney infections, and immune-mediated rashes, including Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN), exfoliative dermatitis, and bullous pemphigoid.

[0020] Specifically, human clinical trials have found that cancers that are highly immunogenic, e.g., hot tumors (as defined in Galon, J., and Bruni, D., Approaches to treat immune hot, altered, and cold tumors with combination immunotherapies, supra, incorporated herein by reference and further described below), high IFN-γ expression, or other acceptable indicators of immunogenic susceptibility, can have improved progression-free survival and / or overall survival when treated with chemotherapy that elicits immune-mediated responses, including, but not limited to, immunogenic cell death and / or regulatory T cell (Treg cell) suppression, in combination with a short-acting CDK4 / 6 inhibitor administered at least prior to chemotherapy administration and / or administered both prior to and concurrently with chemotherapy. When cancer therapy includes these three components in an appropriate dosing regimen, progression-free survival and / or overall survival are enhanced by shifting the T cell environment from an immunosuppressive environment (i.e., Treg cells) to enhanced T cell activity and an increase in cytotoxic T cells (CD8+ cells). In some embodiments, the CDK4 / 6 inhibitor is further administered in a maintenance treatment regimen, where the CDK4 / 6 inhibitor is administered as a single agent without chemotherapy, at regular intervals after completion of chemotherapy treatment, for example, but not limited to, once per week, once every two weeks, once every three weeks, once per month, or once per six weeks. In some embodiments, the CDK4 / 6 inhibitor is further administered in a maintenance treatment regimen with a chemotherapy drug, where the CDK4 / 6 inhibitor is administered in conjunction with a lower dose of chemotherapy, at regular intervals after completion of the initial chemotherapy treatment regimen, for example, but not limited to, once per week, once every two weeks, once every three weeks, once per month, once per six weeks, once per two months, once per three months, once per four months, once per five months, or once per six months.

[0021] In another embodiment, cancers classified as elimination-altered or immunosuppressive-altered according to the scoring system of Galon et al. may show improved progression-free survival and / or overall survival when treated with chemotherapy that enhances immune-mediated anti-tumor responses, including but not limited to chemotherapy that induces immunogenic cell death, in combination with a short-acting CDK4 / 6 inhibitor administered at least prior to chemotherapy, or alternatively both prior to chemotherapy and chemotherapy. In some embodiments, the CDK4 / 6 inhibitor is further administered in a maintenance treatment regimen, where the CDK4 / 6 inhibitor is administered as a single agent without chemotherapy, or administered periodically after completion of chemotherapy treatment, for example, but not limited to, once weekly, once every 2 weeks, once every 3 weeks, once monthly, or once every 6 weeks. In some embodiments, the CDK4 / 6 inhibitor is further administered along with a chemotherapy agent in a maintenance treatment regimen, where the CDK4 / 6 inhibitor is administered along with a lower dose of chemotherapy on a regular basis after completion of the initial chemotherapy administration regimen, for example, but not limited to, once per week, once every two weeks, once every three weeks, once per month, once per six weeks, once per two months, once per three months, once per four months, once per five months, or once per six months.

[0022] In certain embodiments, the short-acting CDK4 / 6 inhibitor is [ka] [In the formula, R is C(H)X, NX, C(H)Y, or C(X)2; X is a linear, branched, or cyclic C1-C5 alkyl group, such as methyl, ethyl, propyl, cyclopropyl, isopropyl, butyl, sec-butyl, tert-butyl, isobutyl, cyclobutyl, pentyl, isopentyl, neopentyl, tert-pentyl, sec-pentyl, and cyclopentyl; Y is NR1R2, R1 and R2 are independently X, or R1 and R2 together are an alkyl group that forms a bridge containing one or two heteroatoms (N, O, or S); Two X groups may together form an alkyl bridge or a bridge containing one or two heteroatoms (N, S, or O) to form a spiro compound. or a pharmaceutically acceptable salt thereof.

[0023] Cytotoxic chemotherapy generally does not discriminate between replicating healthy cells and cancer cells (killing both indiscriminately), including the critical stem cells in the bone marrow that produce white blood cells, red blood cells, and platelets. This chemotherapy-induced bone marrow damage is known as myelosuppression. As white blood cells, red blood cells, and platelets become depleted, patients receiving chemotherapy face increased risk of infection, anemia, fatigue, and bleeding. Myelosuppression often necessitates the administration of rescue interventions, such as growth factors and blood or platelet transfusions, which can lead to delays and reductions in chemotherapy administration. It also results in increased hospital and physician visits, burdening both patients and the healthcare system and increasing patient risk. Myeloprotective agents are drugs that protect hematopoietic stem cells, white blood cells, red blood cells, and / or platelets in situations (e.g., chemotherapy) in which these cells would otherwise be stressed, damaged, or killed.

[0024] Compound I, also known as "trilaciclib" and developed by G1 Therapeutics, Inc., is currently being investigated in a number of human clinical trials for parenteral use as a myeloprotectant administered via intravenous infusion prior to chemotherapy with 1) gemcitabine and carboplatin in metastatic triple-negative breast cancer (mTNBC), 2) topotecan in advanced stage small cell lung cancer (SCLC), 3) carboplatin and etoposide in SCLC, and 4) carboplatin, etoposide, and the PD-L1 immune checkpoint inhibitor atezolizumab (Tecentriq®) in SCLC.

[0025] Compound III, also known as "relociclib" and developed by G1 Therapeutics, Inc., is currently being evaluated for the treatment of EGFR-mutated non-small cell lung cancer in combination with the EGFR inhibitor osimertinib (Tagrisso®), 1) and 2) for the treatment of EGFR-mutated non-small cell lung cancer. ) It is being investigated in a number of human clinical trials as an anti-cancer agent, generally via continuous administration, such as once daily (with breaks as needed at the discretion of the healthcare provider), in combination with fulvestrant to treat ER+, HER2- breast cancer.

[0026] As provided herein, the following examples and discussion are presented using trilaciclib or a pharmaceutically acceptable salt thereof as an exemplary compound. In other embodiments, other short-acting CDK4 / 6 inhibitors than those described above, including, for example, relociclib, can be used. In yet other embodiments, palbociclib or other selective CDK4 / 6 inhibitors, such as abemaciclib or ribociclib, are used. This does not imply that any of these compounds are considered to be alternative embodiments with performance or efficacy comparable to trilaciclib, but potentially with different therapeutic effects, dosages, or outcomes.

[0027] Surprisingly, human clinical trials using trilaciclib as a myeloprotective agent to protect the viability of hematopoietic progenitor and stem cells during chemotherapy have been found to actually improve overall survival in the entire patient population with statistical significance in triple-negative breast cancer (TNBC).Therefore, it is completely unexpected that this human clinical trial showed a better outcome than planned and expected.As shown in Examples 2, 3, and 4, this effect becomes even greater when the immunogenicity of individual tumors is taken into account.This unexpected immuno-oncological effect is the basis of the present invention.

[0028] In contrast, when used in combination with etoposide and carboplatin as a myeloprotective agent to treat small cell lung cancer, which is generally considered immunologically cold and therefore not favorable for inducing an immune response, trilaciclib worked as expected with statistically significant myeloprotective effects, but without statistically significant improvements in progression-free survival or overall survival in the patient population. However, review of the clinical trial data showed that within a subpopulation of responders, patients receiving trilaciclib exhibited significant immune activity, most notably the expansion of new T cell clones (Example 5, Figures 11-14). Importantly, these same patients who exhibited increased T cell clonal expansion also exhibited improved overall survival.

[0029] The nonclinical and clinical data presented herein demonstrate that the antitumor efficacy benefit of trilaciclib is an immune-mediated phenomenon, and that both chemotherapy type and tumor type are associated with outcome. Immune-mediated responses, such as chemotherapy that induces immunogenic cell death, and tumors with a microenvironment more favorable to immune modulation, support the antitumor efficacy of trilaciclib.

[0030] Furthermore, clinical data indicate that factors such as IFN-γ signaling and related biology such as T cell lytic activity, antigen presentation, and chemokine production play prominent roles in the antitumor efficacy of trilaciclib. Importantly, as described herein, factors determining the potential efficacy of CDK4 / 6 antitumor efficacy can be measured prior to the initiation of therapy, providing a valid and reproducible determination of the potential efficacy and implementation of treatment regimens that may extend overall survival and / or progression-free survival.

[0031] For example, SCLC is characterized by high genomic instability and a smoking-associated mutational profile, but SCLC tumors also exhibit significantly lower levels of both major histocompatibility complex class I and class II complexes, a known method of evading antitumor immunity (making them immunologically "cold-like") (Semenova et al., Origins, genetic landscape, and emerging therapies of small cell lung cancer. Genes Dev 2015; 29: 1447-62). Thus, in SCLC, trilaciclib acts to attenuate chemotherapy-induced myelosuppression without necessarily improving antitumor efficacy in this patient population. In contrast, TNBC is generally genomically unstable, and when treated with the potent ICD drug gemcitabine, the tumor microenvironment becomes more immunogenic or "hot-like" (see, e.g., Park et al., How shall we treat early triple-negative breast cancer (TNBC): from the current standard to upcoming immuno-molecular strategies. ESMO Open 2018; 3 (suppl 1): e000357), which may result in improved antitumor efficacy and prolonged overall survival.

[0032] Specifically, as described in the Examples below, at the initial data cutoff date of May 15, 2019, the addition of trilaciclib to a gemcitabine / carboplatin (GC) schedule (both administration schedules) for treating mTNBC established a clinically meaningful improvement in antitumor efficacy compared with GC alone. Notably, the initial data cutoff demonstrated significant improvements in median overall survival (OS) from 12.6 months with GC alone (Group 1: G / C therapy (days 1 and 8 of a 21-day cycle)) to 20.1 months with the addition of trilaciclib to GC (Group 2: G / C therapy (days 1 and 8) + trilaciclib administered intravenously on days 1 and 8 of a 21-day cycle) and 17.8 months with the addition of trilaciclib to GC (Group 3: G / C therapy (days 2 and 9) + trilaciclib administered intravenously on days 1, 2, 8, and 9 of a 21-day cycle) (Table 5; see Figure 2). At the follow-up data cutoff date of May 15, 2020, median overall survival (OS) (95% CI) was 12.6 (6.3, 15.6) months in group 1, not reached (NR) in group 2 (due to prolonged survival in this patient population) (10.2, NR) (HR=0.31, P=0.0016), and 17.8 (12.9, 32.7) months in group 3 (HR=0.40, P=0.0004). For groups 2 and 3 combined, median OS was 19.8 (14.0, NR) months (HR=0.37, P<0.0001 vs. group 1). Importantly, there were no differences in overall response rate (ORR), progression-free survival (PFS), or OS between tumors classified as CDK4 / 6 replication-independent or indeterminate.

[0033] The median overall survival with gemcitabine / carboplatin (GC) alone is consistent with the published literature for mTNBC patients treated in a similar setting (see O'Shaughnessy et al., Phase III study of iniparib plus gemcitabine and carboplatin versus gemcitabine and carboplatin in patients with metastatic triple-negative breast cancer. J Clin Oncol 2014; 32: 3840-47). In a phase III study of iniparib and GC combination versus GC alone in patients who had received 0 to 2 prior chemotherapy regimens for metastatic disease, the median overall survival of 258 patients treated with GC alone was 11.1 months (ibid.). Similarly, in a recent trial of combination chemotherapy for the first-line treatment of patients with mTNBC, the median OS was 12.1 months with GC (Yardley et al., nab-Paclitaxel plus carboplatin or gemcitabine versus gemcitabine plus carboplatin as first-line treatment of patients with triple-negative metastatic breast cancer: results from the tnAcity trial. Ann Oncol 2018; 29: 1763-70).

[0034] The combination of trilaciclib with certain tumors and chemotherapy regimens is thought to enhance immune activation and promote antitumor immunity by differentially shutting down cytotoxic and regulatory T cell subsets in tumors and then recovering cytotoxic T lymphocytes (CTLs) more rapidly than regulatory T cells (Tregs). This differential change in cell cycle rate between CTLs and Tregs increases the ratio of CTLs to Tregs, leading to enhanced T cell activation and reduced Treg-mediated immunosuppressive function. Together, these events promote CTL-mediated clearance of tumor cells. Therefore, the antitumor effects of trilaciclib are due to the temporary proliferation arrest of T cells (protecting them from chemotherapy-induced damage) and the subsequent activation of CTLs in the tumor microenvironment, where Tregs are scarce.

[0035] In addition, T cell receptor (TCR) analysis indicates that trilaciclib may play an important role in the expansion of anti-tumor T cell subsets during treatment. 5 As further described in

[2014] , patients with small cell lung cancer receiving etoposide, carboplatin, and a PD-L1 inhibitor (atezolizumab) (E / P / A) who received trilaciclib had significantly higher numbers of proliferating T cell clones after trilaciclib treatment than patients receiving E / P / A alone (P = 0.01, Figure 11). Additionally, the patient cohort who responded to trilaciclib had more T cell clonal expansion than patients who received placebo (P = 0.001) and more clonal expansion than patients who did not respond to trilaciclib (P = 0.006). Unlike placebo, trilaciclib significantly increased the number and proportion of newly proliferating clones, indicating that the addition of trilaciclib to an etoposide, carboplatin, and atezolizumab treatment regimen enhances T cell-mediated antitumor responses. These data support the induction of immune-mediated responses by trilaciclib.

[0036] Importantly, the ability to extend overall survival in certain tumor types can be predicted prior to administration. For example, as described in Example 2 below, patients receiving trilaciclib whose TNBC was classified as C2 IFN-γ dominant according to the 6-class immune signature classification system of Thorsson et al. (defined in Thorsson et al., "The Immune Landscape of Cancer," supra, which is incorporated herein by reference and further described below) showed statistically significant improvements in overall survival and progression-free survival compared to TNBC patients classified as C2 IFN-γ dominant who did not receive trilaciclib. As described in Example 3, patients receiving trilaciclib whose TNBC had a high "IFN-γ signature" and "expanded immune signature" score according to the classification system of Ayers et al. (defined in Ayers et al., "IFN-γ-related mRNA profile predicts clinical response to PD-1 blockade," supra, which is incorporated herein by reference and further described below) had a high "IFN-γ signature" and "expanded immune signature" score and received trilaciclib. Not yet Similar statistically significant improvements in overall survival and progression-free survival were observed compared to TNBC patients. Furthermore, as described in Example 4, patients with TNBC PD-L1-positive tumors receiving trilaciclib had significantly longer overall survival than patients with TNBC PD-L1-positive tumors not receiving trilaciclib.

[0037] In addition to the immune-stimulating effects of transient CDK4 / 6 inhibition, these effects were found to be independent of tumor CDK4 / 6 replication dependency (see Tables 6-8, below). For example, although mTNBC is primarily a functionally CDK4 / 6 replication-independent disease, some patients enrolled in this human clinical trial described below had CDK4 / 6 replication-dependent tumors. Based on findings from preclinical studies in which palbociclib was administered in combination with carboplatin in an Rb-competent mouse model (Roberts et al., Multiple roles of cyclin-dependent kinase 4 / 6 inhibitors in cancer therapy. J Natl Cancer Inst 2012;104:476-87), the risk of inducing G1 arrest may reduce tumor cell proliferation and negatively impact the efficacy of chemotherapy in CDK4 / 6 replication-dependent tumors. However, preclinical studies of this particular class of CDK4 / 6 inhibitors administered concomitantly with various chemotherapy agents in multiple CDK4 / 6-dependent mouse models, together with clinical data from this study using an established signature of CDK4 / 6 replication dependence (see Table 6), provide no evidence that the short-acting CDK4 / 6 inhibitors described herein negatively affect the antitumor activity of chemotherapy.

[0038] Thus, as provided herein, the inclusion of a CDK4 / 6 inhibitor as described herein in combination with a chemotherapeutic agent that enhances an immune-mediated response, such as, but not limited to, an ICD-inducing chemotherapeutic agent, can be used to treat CDK4 / 6 replication-dependent tumors, CDK4 / 6 replication-independent tumors, or heterogeneous tumors containing both CDK4 / 6-dependent and -independent cells, where the tumor is hot, or in another embodiment, immunosuppressively or excludingly altered. Similarly, inclusion of a CDK4 / 6 inhibitor as described herein in combination with a chemotherapeutic agent, e.g., an ICD-inducing chemotherapeutic agent, can be used to treat CDK4 / 6 replication-dependent tumors, CDK4 / 6 replication-independent tumors, or heterogeneous tumors containing both CDK4 / 6-dependent and -independent cells, which tumors are immunogenic, e.g., are immunogenically hot; have a high immunoscore, e.g., immunoscore I4; are C2 "IFN-γ dominant"; have a high "IFN-γ signature" or "extended immune signature" score; are PD-L1 positive; or are immunogenic as determined by any other recognizable assessment known in the art.

[0039] Thus, in certain embodiments, provided herein is a method for selecting a patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to enhance progression-free survival or overall survival, comprising: The method comprises the steps of: (i) determining whether the cancer has a surrounding microenvironment that is favorable for immune modulation; (ii) determining whether the chemotherapy regimen is capable of inducing an immune-mediated response, e.g., immunogenic cell death (ICD); If both (i) and (ii) are affirmative, then: (iii) administering an effective amount of a CDK4 / 6 inhibitor selected from Compound I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, prior to, or optionally prior to and concurrently with, the administration of said chemotherapy. comprising The methods provided herein are those in which the increase in progression-free survival and / or overall survival is compared to the expected overall survival based on administration of the chemotherapy alone, either based on literature or otherwise published evidence, comparisons in preclinical or clinical trials, or other means recognized by one of skill in the art.

[0040] In some embodiments, determining whether a cancer has a surrounding microenvironment that favors immunomodulation comprises assessing whether it has sufficiently high levels of major histocompatibility complex class I antigens available to elicit an immune effect. In some embodiments, determining whether a cancer has a surrounding microenvironment that favors immunomodulation comprises assessing whether it has sufficiently high levels of major histocompatibility complex class II antigens available to elicit an immune effect. In some embodiments, determining whether a cancer has a surrounding microenvironment that favors immunomodulation comprises assessing whether it has sufficiently high levels of major histocompatibility complex class I and class II antigens available to elicit an immune effect. In some embodiments, the patient has a cancer that is classified as immunogenic. In some embodiments, the patient has a cancer that is classified as hot, as described herein. In some embodiments, the patient has a cancer that is classified as elimination-adapted, as described herein. In some embodiments, the patient has a cancer that is classified as a C2 "IFN-γ dominant" class cancer, as described herein. In some embodiments, the patient has a cancer that is classified as a high "IFN-γ signature" or a high "extended immune signature," as described herein. In some embodiments, the patient has a cancer that is PD-L1 positive.

[0041] In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound II, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound IV, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound V, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor is administered within about 24 hours prior to administration of immune response-mediated chemotherapy, e.g., ICD-induced chemotherapy. In some embodiments, the CDK4 / 6 inhibitor is administered within about 4 hours prior to administration of immune response-mediated chemotherapy, e.g., ICD-induced chemotherapy. In some embodiments, the CDK4 / 6 inhibitor is administered within about 30 minutes prior to administration of immune response-mediated chemotherapy, e.g., ICD-induced chemotherapy. In some embodiments, the CDK4 / 6 inhibitor is first administered about 18 to 28 hours before administration of immune response-mediated chemotherapy, e.g., ICD-guided chemotherapy, and then re-administered about 4 hours before administration of immune response-mediated chemotherapy, e.g., ICD-guided chemotherapy. In some embodiments, the patient is not administered an immune checkpoint inhibitor. In some embodiments, the CDK4 / 6 inhibitor is administered one or more times after completion of chemotherapy treatment in a maintenance treatment regimen, for example, once per week, once every two weeks, once per three weeks, once per month, or once per six months. In some embodiments, the CDK4 / 6 inhibitor is administered in combination with a chemotherapeutic agent one or more times after completion of chemotherapy treatment in a chemotherapy-reduced dose maintenance treatment regimen, for example, at least once per week, at least once per two weeks, at least once per three weeks, at least once per month, at least once per six weeks, at least once per two months, at least once per three months, at least once per four months, at least once per five months, or at least once per six months.

[0042] In another embodiment, there is provided a method for selecting a patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to enhance progression-free survival or overall survival, comprising: The method comprises the steps of: (i) determining whether the cancer is immunogenically susceptible to treatment with the CDK4 / 6 inhibitor; (ii) determining whether the patient can be administered chemotherapy that induces an immune response based on the cancer, e.g., ICD-inducing chemotherapy; (iii) when it is determined that the cancer is immunogenically susceptible to CDK4 / 6 inhibitor treatment and that the chemotherapy that induces an immune response, for example, the ICD-inducing chemotherapy, is administrable, administering an effective amount of the chemotherapy in combination with an effective amount of a short-acting CDK4 / 6 inhibitor selected from Compound I, Compound II, Compound III, Compound IV, or Compound V, or a pharmaceutically acceptable salt thereof. comprising the CDK4 / 6 inhibitor is administered prior to, or prior to and concurrently with, the administration of the chemotherapy; The methods are provided wherein the improvement in progression-free survival and / or overall survival is compared to the progression-free survival and / or overall survival based on administration of chemotherapy alone, either based on literature or otherwise published evidence, comparisons in preclinical or clinical trials, or other means recognized by one of skill in the art.

[0043] In some embodiments, determining whether a cancer is immunogenically susceptible to CDK4 / 6 inhibitor treatment comprises assessing whether it has sufficiently high levels of major histocompatibility complex class I antigens available to elicit an immune effect. In some embodiments, determining whether a cancer is immunogenically susceptible to CDK4 / 6 inhibitor treatment comprises assessing whether it has sufficiently high levels of major histocompatibility complex class II antigens available to elicit an immune effect. In some embodiments, determining whether a cancer is immunogenically susceptible to CDK4 / 6 inhibitor treatment comprises assessing whether it has sufficiently high levels of major histocompatibility complex class I and class II antigens available to elicit an immune effect. In some embodiments, the patient has a cancer classified as immunogenic. In some embodiments, the patient has a cancer classified as hot, as described herein. In some embodiments, the patient has a cancer classified as elimination-adapted, as described herein. In some embodiments, the patient has a cancer classified as a C2 "IFN-γ dominant" class cancer, as described herein. In some embodiments, the patient has a cancer that is classified as a high "IFN-γ signature" or a high "extended immune signature," as described herein. In some embodiments, the patient has a cancer that is PD-L1 positive.

[0044] In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound II, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound IV, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound V, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor is administered within about 24 hours prior to administration of immune response-mediated chemotherapy, e.g., ICD-induced chemotherapy. In some embodiments, the CDK4 / 6 inhibitor is administered within about 4 hours prior to administration of immune response-mediated chemotherapy, e.g., ICD-induced chemotherapy. In some embodiments, the CDK4 / 6 inhibitor is administered within about 30 minutes prior to administration of immune response-mediated chemotherapy, e.g., ICD-induced chemotherapy. In some embodiments, the CDK4 / 6 inhibitor is first administered about 22 to 26 hours before administration of immune response-mediated chemotherapy, e.g., ICD-guided chemotherapy, and then re-administered about 4 hours before administration of immune response-mediated chemotherapy, e.g., ICD-guided chemotherapy. In some embodiments, the patient is not administered an immune checkpoint inhibitor. In some embodiments, the CDK4 / 6 inhibitor is administered one or more times after completion of chemotherapy treatment in a maintenance treatment regimen, e.g., once per week, once every two weeks, once per three weeks, once per month, or once per six months. In some embodiments, the CDK4 / 6 inhibitor is administered in combination with a chemotherapy agent one or more times after completion of chemotherapy treatment in a chemotherapy-reduced dose maintenance treatment regimen, e.g., at least once per week, at least once per two weeks, at least once per three weeks, at least once per month, at least once per two months, at least once per six weeks, at least once per three months, at least once per four months, at least once per six months, or at least once per six months.

[0045] Chemotherapeutics capable of inducing an immune-mediated response are generally known in the art and include, but are not limited to, alkylating agents such as cyclophosphamide, trabectedin, temozolomide, melphalan, dacarbazine, and oxaliplatin; antimetabolites such as methotrexate, mitroxantrone, gemcitabine, and 5-fluorouracil (5-FU); bleomycin and anthracyclines (including doxorubicin, daunorubicin, epirubicin, idarubicin, and valrubicin); cytotoxic antibiotics; taxanes such as paclitaxel, cabazitaxel, and docetaxel; topoisomerase inhibitors such as topotecan, irinotecan, and etoposide; platinum compounds such as carboplatin and cisplatin; 26S proteasome subunit inhibitor bortezomib; vinca alkaloids such as vinblastine, vincristine, vindesine, and vinorelbine; diaziquone; mechlorethamine; mitomycin C; fludarabine; cytosine arabinoside; and any combination thereof. In some embodiments, the ICD-guided chemotherapy is selected from idarubicin, epirubicin, doxorubicin, mitoxantrone, oxaliplatin, bortezomib, gemcitabine, and cyclophosphamide, and combinations thereof.

[0046] Although methods are known for determining whether a patient with a particular cancer is a candidate for chemotherapy that can induce an immune response, the effect of CDK4 / 6 inhibitors on such therapy has not been fully explored, particularly without the use of immune checkpoint inhibitors. Considerations include whether the type of cancer being treated is known to respond to a particular chemotherapy drug, whether the patient has already received a chemotherapy drug in the past, and whether the patient's cancer has developed resistance to the chemotherapy or has phenological characteristics that make it ineffective.

[0047] Suitable target cancers for treatment using the currently described methods with CDK4 / 6 inhibitors include tumors that are immunogenic or susceptible to immuno-oncological chemotherapy treatment regimens. In some embodiments, the patient being treated has an immunogenic cancer selected from the group consisting of estrogen receptor (ER)-positive breast cancer, breast cancer including triple-negative breast cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, classical Hodgkin's lymphoma (cHL), bladder cancer, primary mediastinal B-cell lymphoma (PBMCL), diffuse large B-cell lymphoma, urothelial carcinoma, microsatellite instability-high (MSI-H) solid tumors, mismatch repair-deficient (dMMR) solid tumors, gastric or gastroesophageal junction (GEJ) adenocarcinoma, esophageal squamous cell carcinoma, cervical cancer, endometrial carcinoma, cholangiocarcinoma, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, ovarian cancer, anal canal cancer, colorectal cancer, cutaneous melanoma, endometrial carcinoma, and melanoma.

[0048] Thus, the methods provided herein include: A. A method for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to enhance progression-free survival or overall survival, comprising: The method comprises the steps of: (i) determining whether the cancer has a surrounding microenvironment that is favorable for immune modulation; (ii) determining whether the chemotherapy regimen induces an immune-mediated response, e.g., immunogenic cell death; (iii) if both (i) and (ii) are positive, administering an effective amount of a CDK4 / 6 inhibitor selected from Compound I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, prior to administration of said chemotherapy, or optionally prior to and concurrently with said chemotherapy. comprising The method, wherein the prolongation of progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone, either based on literature or otherwise published evidence, comparisons in preclinical or clinical trials, or other means recognized by those skilled in the art. In some embodiments, the patient is not administered a checkpoint inhibitor during the treatment regimen.

[0049] B. A method for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to enhance progression-free survival or overall survival, comprising: The method comprises the steps of: (i) determining the immunogenic classification of said cancer; (ii) determining whether the patient can be administered chemotherapy capable of inducing an immune-mediated response based on the cancer, e.g., ICD-inducing chemotherapy; (iii) when it is determined that the cancer is amenable to the chemotherapy capable of inducing an immune-mediated response, for example, the ICD-inducing chemotherapy, administering an effective amount of the chemotherapy in combination with an effective amount of a short-acting CDK4 / 6 inhibitor selected from Compound I, Compound II, Compound III, Compound IV, or Compound V, or a pharmaceutically acceptable salt thereof; comprising said CDK4 / 6 inhibitor is administered prior to, or optionally prior to and concurrently with, the administration of said chemotherapy; The method, wherein the improvement in progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone, either based on literature or otherwise published evidence, comparisons in preclinical or clinical trials, or other means recognized by those skilled in the art. In some embodiments, the patient is not administered a checkpoint inhibitor during the treatment regimen.

[0050] C. A method for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to enhance progression-free survival or overall survival, comprising: The method comprises the steps of: (i) determining whether the cancer is immunogenically susceptible to treatment with said CDK4 / 6 inhibitor; (ii) determining whether the patient can be administered chemotherapy that induces an immune response based on the cancer, e.g., ICD-inducing chemotherapy; (iii) when it is determined that the cancer is immunogenically susceptible to CDK4 / 6 inhibitor treatment and that the chemotherapy that induces an immune response, for example, the ICD-inducing chemotherapy, is administrable, administering an effective amount of the chemotherapy in combination with an effective amount of a short-acting CDK4 / 6 inhibitor selected from Compound I, Compound II, Compound III, Compound IV, or Compound V, or a pharmaceutically acceptable salt thereof. comprising the CDK4 / 6 inhibitor is administered prior to, or prior to and concurrently with, the administration of the chemotherapy; The method, wherein the improvement in progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone, either based on literature or otherwise published evidence, comparisons in preclinical or clinical trials, or other means recognized by those skilled in the art. In some embodiments, the patient is not administered a checkpoint inhibitor during the treatment regimen.

[0051] D. A method for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to enhance progression-free survival or overall survival, comprising: The method comprises the steps of: (i) determining whether the cancer is immunogenic; (ii) determining whether the patient can be administered chemotherapy that induces an immune response based on the cancer, e.g., ICD-inducing chemotherapy; (iii) when it is determined that the cancer is immunogenic and the chemotherapy that induces an immune response, for example, the ICD-inducing chemotherapy, is administrable, administering an effective amount of the chemotherapy in combination with an effective amount of a short-acting CDK4 / 6 inhibitor selected from Compound I, Compound II, Compound III, Compound IV, or Compound V, or a pharmaceutically acceptable salt thereof. comprising the CDK4 / 6 inhibitor is administered prior to, or prior to and concurrently with, the administration of the chemotherapy; The method, wherein the improvement in progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone, either based on literature or otherwise published evidence, comparisons in preclinical or clinical trials, or other means recognized by those skilled in the art. In some embodiments, the patient is not administered a checkpoint inhibitor during the treatment regimen.

[0052] E. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy in a patient or patient population selected for extending progression-free survival or overall survival, comprising: (i) determining whether the cancer has a surrounding microenvironment that is favorable for immune modulation; (ii) determining whether the chemotherapy regimen induces an immune-mediated response, e.g., immunogenic cell death; (iii) if both (i) and (ii) are positive, administering an effective amount of a CDK4 / 6 inhibitor selected from Compound I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, prior to administration of said chemotherapy, or optionally prior to and concurrently with said chemotherapy. comprising The use, wherein the prolongation of progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone, based either on literature or otherwise published evidence, comparisons in preclinical or clinical trials, or other means recognized by those skilled in the art. In some embodiments, the patient is not administered a checkpoint inhibitor during the treatment regimen.

[0053] F. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy in a patient or patient population selected for prolonging progression-free survival or overall survival, comprising: The selection of the patient or patient population comprises the steps of: (ii) determining whether the patient can be administered chemotherapy capable of inducing an immune-mediated response based on the cancer, e.g., ICD-inducing chemotherapy; (iii) when it is determined that the cancer is amenable to the chemotherapy capable of inducing an immune-mediated response, for example, the ICD-inducing chemotherapy, administering an effective amount of the chemotherapy in combination with an effective amount of a short-acting CDK4 / 6 inhibitor selected from Compound I, Compound II, Compound III, Compound IV, or Compound V, or a pharmaceutically acceptable salt thereof; comprising said CDK4 / 6 inhibitor is administered prior to, or optionally prior to and concurrently with, the administration of said chemotherapy; The use, wherein the improvement in progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone, based either on literature or otherwise published evidence, comparisons in preclinical or clinical trials, or other means recognized by those skilled in the art. In some embodiments, the patient is not administered a checkpoint inhibitor during the treatment regimen.

[0054] G. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy in a patient or patient population selected for prolonging progression-free survival or overall survival, The selection of the patient or patient population comprises the steps of: (i) determining whether the cancer is immunogenically susceptible to treatment with said CDK4 / 6 inhibitor; (ii) determining whether the patient can be administered chemotherapy that induces an immune response based on the cancer, e.g., ICD-inducing chemotherapy; (iii) when it is determined that the cancer is immunogenically susceptible to CDK4 / 6 inhibitor treatment and that the chemotherapy that induces an immune response, for example, the ICD-inducing chemotherapy, is administrable, administering an effective amount of the chemotherapy in combination with an effective amount of a short-acting CDK4 / 6 inhibitor selected from Compound I, Compound II, Compound III, Compound IV, or Compound V, or a pharmaceutically acceptable salt thereof. comprising the CDK4 / 6 inhibitor is administered prior to, or prior to and concurrently with, the administration of the chemotherapy; The use, wherein the improvement in progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone, based either on literature or otherwise published evidence, comparisons in preclinical or clinical trials, or other means recognized by those skilled in the art. In some embodiments, the patient is not administered a checkpoint inhibitor during the treatment regimen.

[0055] H. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy in a patient or patient population selected for prolonging progression-free survival or overall survival, comprising: The selection of the patient or patient population comprises the steps of: (i) determining whether the cancer is immunogenic; (ii) determining whether the patient can be administered chemotherapy that induces an immune response based on the cancer, e.g., ICD-inducing chemotherapy; (iii) when it is determined that the cancer is immunogenic and the chemotherapy that induces an immune response, for example, the ICD-inducing chemotherapy, is administrable, administering an effective amount of the chemotherapy in combination with an effective amount of a short-acting CDK4 / 6 inhibitor selected from Compound I, Compound II, Compound III, Compound IV, or Compound V, or a pharmaceutically acceptable salt thereof. comprising the CDK4 / 6 inhibitor is administered prior to, or prior to and concurrently with, the administration of the chemotherapy; The use, wherein the improvement in progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone, based either on literature or otherwise published evidence, comparisons in preclinical or clinical trials, or other means recognized by those skilled in the art. In some embodiments, the patient is not administered a checkpoint inhibitor during the treatment regimen. [Brief explanation of the drawings]

[0056] [Figure 1]Figure 1 shows a clinical trial overview of the G1T28-04 human clinical trial evaluating the clinical benefit of trilaciclib (Compound I) in protecting the bone marrow and immune system and enhancing the antitumor efficacy of chemotherapy when administered prior to carboplatin and gemcitabine (GC therapy) in patients with metastatic triple-negative breast cancer (mTNBC). The treatment phase consisted of 21-day cycles: trilaciclib was administered intravenously at 240 mg / m² before the gemcitabine / carboplatin infusion. Gemcitabine was administered intravenously at 1000 mg / m². Carboplatin was administered intravenously at a dose calculated based on each patient's area under the curve (AUC). Peripheral blood samples were collected for flow cytometry analysis at pre-treatment and day 1 of odd-numbered cycles; at post-treatment visits; and at the first survival follow-up. Tumor assessments were performed every 9 weeks until week 39 and every 12 weeks thereafter. Where indicated, LOT = prior therapy, Trila = trilaciclib, Tox = toxicity, PD = disease progression, WD = withdrawal, DC = discontinuation, PI = investigator, ANC = absolute neutrophil count, IV = intravenous, OS = overall survival, PTV = post-procedure visit, FU = follow-up. The effect of trilaciclib on PFS and OS remained stable between two data snapshots: the data cutoff date of June 28, 2019, and nearly a year later, on May 15, 2020. [Figure 2] Figure 2 is a Kaplan-Meier plot of overall survival for human patients with triple-negative breast cancer in group 1 (gemcitabine plus carboplatin on days 1 and 8 only of a 21-day cycle), group 2 (gemcitabine plus carboplatin plus trilaciclib on days 1 and 8 of a 21-day cycle), and group 3 (gemcitabine plus carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle). The x-axis represents months from randomization and the number of patients at risk. The y-axis represents survival. Overall survival was significantly longer in group 3 compared to group 1 (hazard ratio = 0.40; nominal p-value = 0.0004) and group 2 compared to group 1 (hazard ratio = 0.31; nominal p-value = 0.0016). The data cutoff date was May 15, 2020. [Figure 3]Figure 3 is a Kaplan-Meier plot of progression-free survival for human patients with triple-negative breast cancer in group 1 (gemcitabine + carboplatin only on days 1 and 8 of a 21-day cycle), group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), and group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle). The x-axis represents months from randomization and the number of patients at risk. The y-axis represents progression-free survival. The data cutoff date was May 15, 2020. [Figure 4A] Figure 4A shows a forest plot of overall survival for human patients with triple-negative breast cancer in Group 1 (gemcitabine + carboplatin on days 1 and 8 only of a 21-day cycle), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), and Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle). Data are from the intention-to-treat population; data from Groups 2 and 3 for trilaciclib were combined for predefined subgroup analyses. Acquired triple-negative breast cancer refers to patients with confirmed metastatic triple-negative breast cancer and a prior biopsy showing positivity for estrogen and progesterone receptors or HER2. Data from Groups 2 and 3 for trilaciclib were combined for predefined subgroup analyses. Two-sided p values ​​were obtained using a stratified log-rank test. Hazard ratios between the two treatment groups (trilaciclib vs. gemcitabine / carboplatin only), along with their 95% confidence intervals (CIs), were calculated from Cox proportional hazards models in which treatment and appropriate stratification factors were included as fixed terms. Where indicated in the plots: ECOG = Eastern Cooperative Oncology Group. P values ​​were obtained from stratified log-rank tests using appropriate stratification factors as covariates. Analyses were performed using a data cutoff date of June 28, 2019. [Figure 4B]Figure 4B shows a forest plot of progression-free survival for human patients with triple-negative breast cancer in Group 1 (gemcitabine + carboplatin on days 1 and 8 only of a 21-day cycle), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), and Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle). Data are from the intention-to-treat population; data from Groups 2 and 3 for trilaciclib were combined for predefined subgroup analyses. Acquired triple-negative breast cancer refers to patients with confirmed metastatic triple-negative breast cancer whose previous biopsy showed estrogen and progesterone receptor or HER2 positivity. Data from Groups 2 and 3 for trilaciclib were combined for predefined subgroup analyses. Two-sided p values ​​were obtained using a stratified log-rank test. Hazard ratios between the two treatment groups (trilaciclib vs. gemcitabine / carboplatin only), along with their 95% confidence intervals (CIs), were calculated from Cox proportional hazards models in which treatment and appropriate stratification factors were included as fixed terms. Where indicated in the plots: ECOG = Eastern Cooperative Oncology Group. P values ​​were obtained from stratified log-rank tests using appropriate stratification factors as covariates. Analyses were performed using a data cutoff date of June 28, 2019. [Figure 4C]Figure 4C shows a forest plot of overall survival for human patients with triple-negative breast cancer in Group 1 (gemcitabine + carboplatin on days 1 and 8 only of a 21-day cycle), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), and Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle). Data are from the intention-to-treat population; data from Groups 2 and 3 for trilaciclib were combined for predefined subgroup analyses. Acquired triple-negative breast cancer refers to patients with confirmed metastatic triple-negative breast cancer whose previous biopsy showed estrogen and progesterone receptor or HER2 positivity. Data from Groups 2 and 3 for trilaciclib were combined for predefined subgroup analyses. Two-sided p values ​​were obtained using a stratified log-rank test. Hazard ratios between the two treatment groups (trilaciclib vs. gemcitabine / carboplatin only), along with their 95% confidence intervals (CIs), were calculated from Cox proportional hazards models in which treatment and appropriate stratification factors were included as fixed terms. Where indicated in the plots: ECOG = Eastern Cooperative Oncology Group. P values ​​were obtained from stratified log-rank tests using appropriate stratification factors as covariates. Analyses were performed using a data cutoff date of May 15, 2020. [Figure 4D]Figure 4D shows a forest plot of progression-free survival for human patients with triple-negative breast cancer in Group 1 (gemcitabine + carboplatin on days 1 and 8 only of a 21-day cycle), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), and Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle). Data are from the intention-to-treat population; data from Groups 2 and 3 for trilaciclib were combined for predefined subgroup analyses. Acquired triple-negative breast cancer refers to patients with confirmed metastatic triple-negative breast cancer whose previous biopsy showed estrogen and progesterone receptor or HER2 positivity. Data from Groups 2 and 3 for trilaciclib were combined for predefined subgroup analyses. Two-sided p values ​​were obtained using a stratified log-rank test. Hazard ratios (HRs) between the two treatment groups (trilaciclib vs. gemcitabine / carboplatin only), along with their 95% confidence intervals (CIs), were calculated from Cox proportional hazards models in which treatment and appropriate stratification factors were included as fixed terms. Where indicated in the plots: ECOG = Eastern Cooperative Oncology Group. P values ​​were obtained from stratified log-rank tests using appropriate stratification factors as covariates. Analyses were performed using a data cutoff date of May 15, 2020. [Figure 5]Figure 5 shows the normalized mean frequency of the interferon-gamma (IFN-γ)-positive CD8+ T cell population (IFN-γ+IL-17A-[CD3+CD8+]) after ex vivo stimulation. Only patients who received three or more cycles of gemcitabine / carboplatin (GC) were included in the analysis (n = 8–15 per cohort), and statistical outliers were excluded. Error bars represent 95% confidence intervals. Group 1 (gemcitabine + carboplatin on days 1 and 8 of a 21-day cycle only), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), and Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle). Data points represent samples taken at different time points from the same patient. Where indicated in the graph: C = cycle; D = day. [Figure 6]Figure 6 is from Galon, J., and Bruni, D., Approaches to treat immune hot, altered, and cold tumors with combination immunotherapies, Nature Reviews Drug Discovery (18), March 2019, the entire contents of which are incorporated herein by reference.This is a facsimile of Figure 3 shown on pages 197-218, illustrating an immunogram for use as a tool to guide anticancer therapy. Cancers can be classified into four major subtypes (hot, altered elimination, altered immunosuppressive, and cold) according to the presence and distribution of their associated T cells (CD3+ and CD8+). Hot cancers are defined by the simultaneous presence of immune component parameters: cell type (CD3+, CD8+, follicular helper T (TFH), T helper 1 (TH1), memory and exhausted T cells); location (tumor leading edge, tumor center, and tertiary lymphoid structures); density (immune density and quantity); and functional immune orientation (chemokines, cytokines, cytotoxic factors, adhesion, attraction, and TH1). As shown on the immunogram, DORA2A: A2A adenosine receptor, βm: β2-microglobulin, BET: bromodomain and extra-terminal motif protein, BTLA: B and T lymphocyte attenuator, CAR T cell: chimeric antigen receptor T cell, CCR: CC-chemokine receptor, CIN: chromosomal instability, CSF1R: colony-stimulating factor 1 receptor, CTLA4: cytotoxic T lymphocyte-associated antigen, CXCL: CXC-chemokine ligand, DDR: DNA damage response, ECM: extracellular matrix, EMT: epithelial-mesenchymal transition, FDA: Food and Drug Administration, FGFR3: fibroblast growth factor receptor 3, FOXP3: forkhead box P3, GITR: glucocorticoid-induced TNFR-related protein, GM-CSF: granulocyte-macrophage colony-stimulating factor, HDAC: histone deacetylase, HIF1α: hypoxia-inducible factor 1-α, HLA: human leukocyte antigen, HMA: demethylating agent, IAP: inhibitor of apoptosis family (also known as XIAP), ICAM1: intercellular adhesion molecule 1, ICD: immunogenic cell death, ICOS: inducible T-cell costimulator, ICP: immune checkpoint, IDO: indoleamine 23-dioxygenase, IFN: interferon, IL: interleukin, LAG3: lymphocyte activation gene 3, LIGHT: tumor necrosis factor superfamily member 14, MAdCAM1: mucosal addressin cell adhesion molecule 1, MCL1: inducible myeloid leukemia cell differentiation protein Mcl1, MDSC: myeloid-derived suppressor cell, MEK: mitogen-activated protein kinase kinase, MET: mesenchymal-epithelial transition, MSI: microsatellite instability, NK: natural killer, NOS1: nitric oxide synthase 1, PD-1: programmed cell death protein 1, PD-L1: PD-1 ligand, PI3Kγ: phosphoinositide 3-kinase-γ, PPARγ: peptide oxisome proliferator-activated receptor-γ, SIGLEC9: sialic acid-binding Ig-like lectin 9, STING: stimulator of interferon genes, TDO: tryptophan 2,3-dioxygenase, TGFβ: transforming growth factor-β, TIGIT: T-cell immunoglobulin and ITIM domain, TIM3: T-cell immunoglobulin and mucin domain containing 3, TKI: tyrosine kinase inhibitor, TLR: Toll-like receptor, Treg cells: regulatory T cells, VCAM1: vascular cell adhesion molecule 1, VEGF: vascular endothelial growth factor, VISTA: V-domain Ig inhibitor of T-cell activation, XCL1: lymphotactin, XCR1: chemokine XCR receptor 1. A lowercase "i" following an acronym or abbreviation indicates an inhibitor; a lowercase "a" following an acronym or abbreviation indicates an agonist. [Figure 7]Figure 7A is a reproduction of Figure 1A shown in Galon, J., and Bruni, D., Approaches to Treat Immune Hot, Altered, and Cold Tumors with Combination Immunotherapies, Nature Reviews Drug Discovery (18), March 2019, pp. 197-218, the entire contents of which are incorporated herein by reference, illustrating examples of hot, altered, and cold immune cancers. Dark (3,3'-diaminobenzidine (DAB)) staining indicates CD3+ T cells, and light (alkaline phosphatase) counterstaining indicates homogeneous tissue background staining. The level and spatial distribution of CD3+ and CD8+ T cell infiltration distinguishes four distinct solid tumor phenotypes: hot (or inflammatory); altered, which can be eliminative or immunosuppressive; and cold (or noninflammatory). These tumor phenotypes are characterized by high, intermediate, and low immunoscores, respectively. Figure 7B is a reproduction of Figure 1B shown in Galon, J., and Bruni, D., Approaches to Treat Immune Hot, Altered, and Cold Tumors with Combination Immunotherapies, Nature Reviews Drug Discovery (18), March 2019, pp. 197-218, the entire contents of which are incorporated herein by reference. It shows a schematic diagram of the four subtypes of immune tumors. Of note, in tumors with immune-suppressive transformation, CD3+ and CD8+ T cell infiltration is low in the tumor center and high in the tumor leading edge, resulting in an overall intermediate immunoscore. In contrast, tumors with immune-suppressive transformation show a more uniform pattern of CD3+ and CD8+ T cell infiltration (low). CT, tumor center; Hi, high; IM, tumor leading edge; Lo, low. [Figure 8A]Figure 8A shows the Ayers IFN-γ signature score distribution plot for pretreatment cancer samples from patients participating in the G1T28-04 clinical trial (NCT02978716). The x-axis represents the probability density curve, and the y-axis represents the calculated Ayers IFN-γ signature score. The dotted vertical line on the left side of the graph represents the first tertile, the dotted vertical line on the right side of the graph represents the second tertile, and the dotted vertical line between the first and second tertiles represents the median. [Figure 8B] Figure 8B shows the Ayers' extended immune signature score distribution plot for pretreatment cancer samples from patients participating in the G1T28-04 clinical trial (NCT02978716). The x-axis represents the probability density curve, and the y-axis represents the calculated Ayers' IFN-γ signature score. The dotted vertical line on the left side of the graph represents the first tertile, the dotted vertical line on the right side of the graph represents the second tertile, and the dotted vertical line between the first and second tertiles represents the median. [Figure 8C] Figure 8C is a Kaplan-Meier plot of overall survival for human patients with triple-negative breast cancer in Group 1 (gemcitabine + carboplatin on days 1 and 8 only of a 21-day cycle), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle), and Group 4 (Group 2 + Group 3), who were determined to have a high Ayers IFN-γ signature score in the G1T28-04 clinical trial (NCT02978716). The x-axis represents months from randomization. The y-axis represents the survival rate. Overall survival was significantly longer in Group 4 (Group 2 + 3) compared with Group 1 (p = 0.0194). Legend: dashed line (- - - -) group 1; dotted line (····) group 2; solid line (-) group 3; dash-dotted line (-·-·-) group 4. [Figure 8D]Figure 8D is a Kaplan-Meier plot of overall survival for human triple-negative breast cancer patients in Group 1 (gemcitabine + carboplatin on days 1 and 8 only of a 21-day cycle), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle), and Group 4 (Group 2 + Group 3), who were determined to have a low Ayers IFN-γ signature score in the G1T28-04 clinical trial (NCT02978716). The x-axis represents months from randomization. The y-axis represents the percentage of survival. Legend: dashed line (- - - -) group 1; dotted line (····) group 2; solid line (-) group 3; dash-dotted line (-·-·-) group 4. [Figure 8E] Figure 8E is a Kaplan-Meier plot of progression-free survival (PFS) for human triple-negative breast cancer patients in Group 1 (gemcitabine + carboplatin on days 1 and 8 of a 21-day cycle only), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle), and Group 4 (Group 2 + Group 3), who were determined to have a low Ayers IFN-γ signature score in the G1T28-04 clinical trial. The x-axis represents months from randomization. The y-axis represents survival. Legend: Dashed line (- - - -) Group 1; dotted line (····) Group 2; solid line (-) Group 3; dashed-dotted line (-·-··) Group 4. [Figure 8F]Figure 8F is a Kaplan-Meier plot of progression-free survival (PFS) for human patients with triple-negative breast cancer in the G1T28-04 clinical trial, who were assigned to Group 1 (gemcitabine + carboplatin on days 1 and 8 of a 21-day cycle only), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle), and Group 4 (Group 2 + Group 3). The x-axis represents months from randomization. The y-axis represents survival. Legend: Dashed line (- - - -) Group 1; dotted line (····) Group 2; solid line (-) Group 3; dashed-dotted line (-·-··) Group 4. [Figure 9A] Figure 9A is a Kaplan-Meier plot of overall survival for human patients with triple-negative breast cancer in the G1T28-04 clinical trial, who were determined to have a high Ayers extended immune signature score. The patients were divided into groups: Group 1 (gemcitabine + carboplatin on days 1 and 8 of a 21-day cycle only), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle), and Group 4 (Group 2 + Group 3). The x-axis represents months from randomization. The y-axis represents the survival rate. Overall survival was significantly longer in Group 4 (Group 2 + 3) compared with Group 1 (p = 0.0266). Legend: dashed line (- - - -) group 1; dotted line (····) group 2; solid line (-) group 3; dash-dotted line (-·-·-) group 4. [Figure 9B]Figure 9B is a Kaplan-Meier plot of overall survival for human patients with triple-negative breast cancer in the G1T28-04 clinical trial, who were assigned to Group 1 (gemcitabine + carboplatin on days 1 and 8 of a 21-day cycle only), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle), and Group 4 (Group 2 + Group 3). The x-axis represents months from randomization. The y-axis represents survival. Legend: Dashed line (- - - -) Group 1; dotted line (····) Group 2; solid line (-) Group 3; dashed-dotted line (-·-··) Group 4. [Figure 9C] Figure 9C is a Kaplan-Meier plot of progression-free survival (PFS) for human triple-negative breast cancer patients in Group 1 (gemcitabine + carboplatin on days 1 and 8 only of a 21-day cycle), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle), and Group 4 (Group 2 + Group 3), who were determined to have a high Ayers extended immune signature score in the G1T28-04 clinical trial. The x-axis represents months from randomization. The y-axis represents survival. Legend: Dashed line (- - - -) Group 1; dotted line (····) Group 2; solid line (-) Group 3; dashed-dotted line (-·-··) Group 4. [Figure 9D]Figure 9D is a Kaplan-Meier plot of progression-free survival (PFS) for human triple-negative breast cancer patients in the G1T28-04 clinical trial, who were assigned to Group 1 (gemcitabine + carboplatin on days 1 and 8 of a 21-day cycle only), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle), and Group 4 (Group 2 + Group 3). The x-axis represents months since randomization. The y-axis represents survival. Legend: Dashed line (- - - -) Group 1; dotted line (····) Group 2; solid line (-) Group 3; dashed-dotted line (-·-··) Group 4. [Figure 10A] Figure 10A is a Kaplan-Meier plot of overall survival for human patients with triple-negative breast cancer who were assigned to Group 1 (gemcitabine + carboplatin on days 1 and 8 only of a 21-day cycle), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle), and Group 4 (Group 2 + Group 3), who were determined to have a C2 IFN-γ-dominant 6-class immune signature in the G1T28-04 clinical trial. The x-axis represents months from randomization. The y-axis represents the survival rate. Overall survival was significantly longer in Group 4 (Group 2 + 3) compared with Group 1 (p = 0.036). Legend: dashed line (- - - -) group 1; dotted line (····) group 2; solid line (-) group 3; dash-dotted line (-·-·-) group 4. [Figure 10B]Figure 10B is a Kaplan-Meier plot of overall survival for human patients with triple-negative breast cancer in the G1T28-04 clinical trial, who were classified as C2 (not IFN-γ dominant) by the six-class immune signature score. The following groups were treated: Group 1 (gemcitabine + carboplatin on days 1 and 8 of a 21-day cycle only), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9 of a 21-day cycle), and Group 4 (Group 2 + Group 3). The x-axis represents months since randomization. The y-axis represents survival. Legend: Dashed line (- - - -) Group 1; dotted line (····) Group 2; solid line (-) Group 3; dashed-dotted line (-·-··) Group 4. [Figure 10C] Figure 10C is a Kaplan-Meier plot of progression-free survival (PFS) for human patients with triple-negative breast cancer who were assigned to Group 1 (gemcitabine + carboplatin on days 1 and 8 only of a 21-day cycle), Group 2 (gemcitabine + carboplatin + trilaciclib on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine + carboplatin on days 2 and 9 of a 21-day cycle, trilaciclib on days 1, 2, 8, and 9), and Group 4 (Group 2 + Group 3), as determined by a six-class immune signature score of C2 IFN-γ dominant in the G1T28-04 clinical trial. The x-axis represents months from randomization. The y-axis represents survival percentage. Legend: dashed line (- - - -) group 1; dotted line (····) group 2; solid line (-) group 3; dash-dotted line (-·-·-) group 4. [Figure 10D]Figure 10D is a Kaplan-Meier plot of progression-free survival (PFS) for human patients with triple-negative breast cancer who were classified as C2 non-IFN-γ dominant by the 6-class immune signature score in the G1T28-04 clinical trial. The x-axis represents months since randomization. The y-axis represents survival. Legend: dashed line (- - - -) group 1; dotted line (····) group 2; solid line (-) group 3; dash-dotted line (-·-·-) group 4. [Figure 11] Figure 11 shows the number of expanded T cell clones measured by differential abundance analysis of T cell receptor β sequences in whole blood of patients receiving trilaciclib or placebo after induction and before the start of maintenance therapy versus baseline (pre-induction). Horizontal bars indicate the median number of expanded clones for each group. [Figure 12] Figure 12 shows the number of proliferating T cell clones measured by differential abundance analysis of T cell receptor β sequences in whole blood of responders and non-responders after induction and before the start of maintenance therapy relative to baseline (pre-induction). Horizontal bars indicate the median number of proliferating clones for each group. [Figure 13] Figure 13 shows the number of newly expanded T cell clones in responders and non-responders receiving placebo or trilaciclib after induction and before the start of maintenance therapy relative to baseline (pre-induction). The horizontal bars indicate the median number of expanded clones in each group. [Figure 14] Figure 14 shows the proportion of newly expanded T cell clones in responders and non-responders receiving placebo or trilaciclib after induction and before the start of maintenance therapy relative to baseline (pre-induction). Horizontal bars indicate the median number of expanded clones in each group. DETAILED DESCRIPTION OF THE INVENTION

[0057] definition Compounds are described using standard nomenclature. Unless otherwise defined, 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.

[0058] The terms "a" and "an" do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item. The term "or" means "and / or." The recitation of ranges of values, unless otherwise stated, is merely intended to serve as a shorthand method of referring individually to each individual value falling within the range, and each individual value is incorporated herein as if it were individually recited herein. The endpoints of all ranges are included within the range and are independently combinable. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of examples or illustrative language (e.g., "etc.") is intended merely to better describe the invention and does not imply a limitation on the scope of the invention unless otherwise claimed. Unless otherwise defined, 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.

[0059] "Effective amount," as used herein, means an amount that provides a therapeutic or prophylactic benefit.

[0060] "Treating" a disease, as the term is used herein, means reducing the frequency or severity of at least one sign or symptom of the disease or disorder suffered by a patient (i.e., palliative treatment) or reducing the cause or effect of the disease or disorder (i.e., disease-modifying treatment).

[0061] As referred to herein, a "host," "subject," "patient," or "individual" treated in accordance with the methods described herein is a mammal, including a human.

[0062] In this disclosure, various aspects of the present invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and is not intended to limit the scope of the present invention. The description of a range should be considered to specifically disclose all possible subranges and individual numerical values ​​within that range. For example, the description of a range such as 1 to 6 should be considered to specifically disclose subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, and individual numerical values ​​within that range, such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This is true regardless of the width of the range.

[0063] As used herein, a "pharmaceutical composition" is a composition comprising 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 into a single dosage form or may be provided together in separate dosage forms with an explanation that the active agents are used together to treat a disorder described herein.

[0064] As used herein, "pharmaceutically acceptable salts" refer to derivatives of the disclosed compounds in which the parent compound is modified by forming its inorganic and organic non-toxic acid or base addition salts. Salts of the present compounds can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid form of these compounds with a stoichiometric amount of an appropriate base (such as hydroxide, carbonate, bicarbonate, Na, Ca, Mg, or K), or by reacting the free base form of these compounds with a stoichiometric amount of an appropriate acid. Such reactions are generally carried out in water or an organic solvent, or a mixture of both. Generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are typical, where practicable. Salts of the present compounds also include solvates of the compounds and of the compound salts.

[0065] Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids. Pharmaceutically acceptable salts include, for example, conventional non-toxic salts of the parent compound formed from non-toxic inorganic or organic acids, and quaternary ammonium salts. For example, conventional non-toxic acid salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, and nitric acid; acetic acid, 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, benzoic acid, salicylic acid, mesylic acid, esylic acid, besylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, HOOC-(CH2) nIncluded are salts made from organic acids such as —COOH, where n is 0 to 4, or with different acids that yield the same counterion. Lists of other suitable salts can be found, for example, in Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton, Pa., p. 1418 (1985).

[0066] The term "carrier" applied to pharmaceutical compositions / combinations of the invention refers to a diluent, excipient, or vehicle with which an active compound is provided.

[0067] As used herein, the term "immune checkpoint inhibitor (ICI)" refers to an inhibitory therapy that targets immune checkpoints, which are important regulators of the immune system that, when stimulated, can dampen the immune response to immune stimulation. Some cancers can protect themselves from attack by stimulating immune checkpoint targets. ICIs block inhibitory checkpoints and restore immune system function. ICIs include immune checkpoint proteins such as programmed cell death-1 protein (PD-1), PD-1 ligand-1 (PD-L1), PD-1 ligand-2 (PD-L2), CTLA-4, LAG-3, TIM-3, and V-domain Ig suppressor of T-cell activation (VISTA), B7-H3 / CD276, indoleamine 2,3-dioxygenase (IDO), killer immunoglobulin-like receptors (KIRs), carcinoembryonic antigen cell adhesion molecules (CEACAMs) such as CEACAM-1, CEACAM-3, and CEACAM-5, sialic acid-binding immunoglobulin-like lectin 15 (Siglec-15), T cell immunoreceptor with Ig and ITIM domains (TIGIT), and B and T lymphocyte attenuator (BTLA) protein. Immune checkpoint inhibitors are known in the art.

[0068] In some embodiments, the term "CDK4 / 6 replication-independent cancer" refers to cancers that do not significantly require CDK4 / 6 activity for replication. Such types of cancer are often, but not always, characterized by (e.g., have cells that exhibit) increased CDK2 activity levels or decreased or absent expression of retinoblastoma tumor suppressor proteins or retinoblastoma family member proteins, such as, but not limited to, p107 and p130. The increased CDK2 activity levels or decreased or absent expression of retinoblastoma tumor suppressor proteins or retinoblastoma family member proteins can be, for example, increased or decreased compared to normal cells. In some embodiments, the increased CDK2 activity levels can be associated with (e.g., can result from or be seen in conjunction with) MYC proto-oncogene amplification or overexpression. In some embodiments, the increased CDK2 activity levels can be associated with overexpression of cyclin E1, cyclin E2, or cyclin A.

[0069] In some embodiments, the term "CDK4 / 6 replication-dependent cancer" refers to cancer that requires CDK4 / 6 activity for replication or proliferation, or that can be inhibited by the activity of selective CDK4 / 6 inhibitors.This type of cancer and disease can be characterized by the presence of functional retinoblastoma (Rb) protein (e.g., have cells that show it).This type of cancer and disease is classified as Rb positive.

[0070] Prediction of anti-cancer / immune effects of CDK4 / 6 inhibitor therapy A number of tumor-related or immune-related biomarkers can be used as predictors of whether or not adding a CDK4 / 6 inhibitor to a chemotherapy regimen, including ICD-guided chemotherapy, will achieve anti-tumor effects. Such predictive biomarkers include tumor cell expression of immunosuppressive molecules (e.g., PD-L1); molecular profiling of the tumor microenvironment, including inflammatory gene expression; assessment of mutational landscape and neoantigen load; mismatch repair deficiency and MSI; tumor aneuploidy; immune infiltration; and immunoscore (see generally, Galon, J., and Bruni, D., Approaches to treat immune hot, altered, and cold tumors with combination immunotherapies, Nature Reviews Drug Discovery (18), March 2019, 197-218, which is incorporated herein by reference). For example, tumors with high levels of somatic copy number alterations (SCNAs) are associated with reduced expression of cytotoxic immune infiltration in melanoma patients (see Davoli et al., Tumor aneuploidy correlates with markers of immune evasion and with reduced response to immunotherapy, Science 355, eaaf8399 (2017)). Patients with tumors with high levels of SCNAs and low expression of cytotoxic immune infiltration are unlikely to achieve the antitumor effects that result in improved overall survival when a CDK4 / 6 inhibitor is added to their chemotherapy regimen.

[0071] Importantly, Galon, J., and Bruni, D., Approaches to Treat Immune Hot, Altered, and Cold Tumors with Combination Immunotherapies (ibid.), describes numerous strategies for treating tumors in a manner that induces improved immune responses, but does not describe the use of CDK4 / 6 inhibitors in combination with ICD-guided chemotherapy to accomplish this (see Figure 6). Galon's article highlights an inventive and unexpected aspect of the present invention, since, despite the comprehensiveness of the literature in prestigious scientific journals, it did not describe or suggest the careful use or appropriate selection of protocols that include CDK4 / 6 inhibitors in immunotherapy as part of an effective anticancer therapy that could extend the progression-free or overall survival of cancer patients. This clinical result is unexpected. Additional parameters, including tumor abnormalities, overall immune status, immune cell infiltration, checkpoint defects, soluble inhibitor defects, inhibitory tumor metabolic defects, and tumor immune effector sensitivity, can also be used to determine whether the anti-tumor effect resulting in extended overall survival can be achieved by adding CDK4 / 6 inhibitors to these chemotherapy regimens. Evaluation of these factors can be achieved by a combination of tumor genomics, immunoscore assays, immunohistochemistry, standard blood assays, and immune gene signatures, both pre- and post-treatment (see Blank et al., The "cancer immunogram," Science 352, 658-660 (2016)).

[0072] Tumor immunogenicity classification As mentioned above, tumors can be classified based on certain immunogenic characteristics. Importantly, it has been found that tumors can progress through various classifications over time. It has also been found that in certain cases, certain tumors can have different immunogenic characteristics in different individuals.

[0073] As demonstrated herein, immunologically hot tumors are those with (i) high T cell and cytotoxic T cell infiltration, i.e., a high immunoscore; and (ii) checkpoint activation (programmed cell death protein 1 (PD-1), cytotoxic T lymphocyte-associated antigen 4 (CTLA4), T cell immunoglobulin mucin receptor 3 (TIM3), and lymphocyte activation gene 3 (LAG3)) or otherwise impaired T cell function (e.g., extracellular potassium-driven T cell suppression). In addition to the presence of tumor-infiltrating lymphocytes (TILs) and expression of anti-programmed cell death ligand 1 (PD-L1) on tumor-associated immune cells, hot tumors characteristically exhibit underlying genomic instability and the presence of a pre-existing anti-tumor immune response. See, for example, Galon, J., and Bruni, D., Approaches to treat immune hot, altered, and cold tumors with combination immunotherapies, Nature Reviews Drug Discovery (18), March 2019, 197-218, the entire contents of which are incorporated herein by reference.

[0074] Tumors that frequently exhibit immunologically hot tumor characteristics include, but are not limited to, bladder cancer, renal cell carcinoma, liver cancer (hepatocellular carcinoma), non-small cell lung cancer, colon adenocarcinoma, breast invasive carcinoma, cholangiocarcinoma, esophageal carcinoma, Merkel cell carcinoma, HPV+ head and neck squamous cell carcinoma, advanced-stage melanoma, cutaneous melanoma, endometrial cancer, gastric cancer, and cervical cancer; Hodgkin lymphoma; diffuse large B-cell lymphoma; and tumors with microsatellite instability (MSI). An exemplary resected tumor with immunologically hot tumor characteristics is shown in Figure 7A.

[0075] Immunosuppressively transformed tumors are characterized by (i) insufficient, or even absent, T cell and cytotoxic T cell infiltration (intermediate immunoscore), (ii) the presence of soluble inhibitory mediators (transforming growth factor-β (TGFβ), interleukin-10 (IL-10), and vascular endothelial growth factor (VEGF)), (iii) the presence of immunosuppressive cells (myeloid-derived suppressor cells and regulatory T cells), and (iv) the presence of T cell checkpoints (PD-1, CTLA4, TIM3, and LAG3). Immunosuppressively transformed tumor sites exhibited low levels of immune infiltration (Figure 7A), suggesting a lack of physical barriers and an immunosuppressive environment that limits further T cell recruitment and proliferation. See, e.g., Galon, J., and Bruni, D., Approaches to treat immune hot, altered, and cold tumors with combination immunotherapies, Nature Reviews Drug Discovery (18), March 2019, 197-218, the entire contents of which are incorporated herein by reference. An exemplary resected tumor with characteristics of an immunologically altered, immunosuppressive tumor is shown in Figure 7A.

[0076] Immunologically transformed tumors are characterized by (i) the absence of T cell infiltration within the tumor bed; T cell accumulation at the tumor margin (tumor leading edge) (intermediate immunoscore), (ii) activation of oncogenic pathways, (iii) epigenetic regulation and reprogramming of the tumor microenvironment, (iii) abnormalities in the tumor vasculature and / or stroma, and (iv) hypoxia. In immunologically transformed tumors, T cells are unable to infiltrate the tumor but are found at the tumor edge (tumor leading edge). This "negative" phenotype reflects the intrinsic ability of the host immune system to efficiently elicit T cell-mediated immune responses and the tumor's ability to evade such responses by physically concealing T cell infiltration (Figure 7A). See, e.g., Galon, J., and Bruni, D., Approaches to treat immune hot, altered, and cold tumors with combination immunotherapies, Nature Reviews Drug Discovery (18), March 2019, 197-218, the entire contents of which are incorporated by reference. An exemplary resected tumor with characteristics of an immunologically altered tumor is shown in Figure 7A.

[0077] Cold tumors are characterized by (i) the absence of T cells within and at the tumor edge (low immunoscore) and (ii) unsuccessful T cell priming (low tumor mutational burden, insufficient antigen presentation, and intrinsic insensitivity to T cell cytotoxicity). Cold tumors may also exhibit low PD-L1 expression. In addition to poor invasion, cold tumors have also been described as immunologically unresponsive (expressing little PD-L1) and characterized by high proliferation accompanied by low mutational burden (low neoantigen expression) and low expression of antigen presentation markers such as major histocompatibility complex class I (MHCI). See, for example, Galon, J., and Bruni, D., Approaches to treat immune hot, altered, and cold tumors with combination immunotherapies, Nature Reviews Drug Discovery (18), March 2019, 197-218, the entire contents of which are incorporated herein by reference. An exemplary resected tumor with cold immune tumor characteristics is shown in Figure 7A.

[0078] Non-immunogenic, or "cold," tumors lack T cell infiltration, a sign that the immune response is not functioning in these tumors. The lack of T cells makes it difficult to induce an immune response with immunotherapeutic agents. The microenvironment surrounding cold tumors contains myeloid-derived suppressor cells (MDSCs) and regulatory T cells (Tregs), which are known to dampen the immune response and inhibit T cells attempting to migrate into the tumor. Other characteristics of cold tumors include the lack of tumor antigens, antigen-presenting blood vessels, the absence of T cell activation, and defective CD8+ homing to the tumor bed.

[0079] These types of tumors are often treated with conventional cancer therapies because checkpoint inhibitor and immunotherapy approaches have not been effective. Some breast, ovarian, prostate, and pancreatic cancers, neuroblastoma, small cell lung cancer, and glioblastoma are generally cold tumors.

[0080] Determination of tumor immunogenicity classification can be performed on resected tumors (primary or metastatic) (see, e.g., Figure 7A). Less invasive diagnostic methods, such as immunopositron emission tomography (PET) imaging to detect intratumoral CD8+ T cells, can also be used. For a description of immunoPET detection of intratumoral CD8+ T cells, see Rooney et al., Molecular and genetic properties of tumors associated with local immune cytolytic activity, Cell 160, 48-61 (2015), which is incorporated by reference.

[0081] Additionally, bulk gene expression profiles can be used to determine the immunogenicity classification of tumors (see Ali et al., Patterns of immune infiltration in breast cancer and their clinical implications: a gene-expression-based retrospective study, PLOS Med. 13, e1002194 (2016); Newman et al., Robust enumeration of cell subsets from tissue expression profiles, Nat. Methods 12, 453-457 (2015); Rooney et al., Molecular and genetic properties of tumors associated with local immune cytolytic activity, Cell 160, 48-61 (2015); Bindea, G. et al., Spatiotemporal dynamics of intratumoral immune cells reveal the immune landscape in human cancer. Immunity 39, 782-795 (2013), each of which is incorporated herein by reference).

[0082] Several additional tools, such as CIBERSORT (which infers the relative proportions of immune subsets within the total leukocyte population), xCell (which predicts immune cell abundance in the entire TME), TIMER (which creates a proportion-based enrichment score for 64 immune and stromal cell types), and integrative immunogenomics (which used a CIBERSORT-based approach to notably identify six immune subtypes of cancer), can be used to assess the abundance of intratumoral immune infiltrates by using deconvolution of bulk gene expression data (Newman et al., Robust enumeration of cell subsets from tissue expression profiles, Nat. Methods 12, 453-457 (2015); Gentles et al., The prognostic landscape of genes and infiltrating immune cells across human cancers, Nat. Med. 21, 938-945 (2015); Aran et al., xCell: digitally portraying the tissue cellular heterogeneity landscape, each of which is incorporated herein by reference). Genome Biol. 18, 220 (2017); Li et al., TIMER: a web server for comprehensive analysis of tumor-infiltrating immune cells, Cancer Res. 77, e108-e110 (2017); Thorsson, V. et al., The immune landscape of cancer. Immunity 48, 812-830 (2018)).

[0083] Immunoscore Immunoscore is a digital pathology immunoassay based on IHC that measures the density of CD3+ and CD8+ T cells in various tumor locations. The Immunoscore scoring system was defined in a large, international, SITC-led retrospective validation study conducted on over 2,500 patients with Stage I-III colon cancer (see Pages et al., International validation of the consensus Immunoscore for the classification of colon cancer: a prognostic and accuracy study, The Lancet, Volume 391, Issue 10135, Pages 2128-2139, May 26, 2018, incorporated herein by reference). Commercially available Immunoscore assays are available, for example, from HalioDx, Inc. (Richmond, Va.). Briefly, CD3 and CD8 immunostained formalin-fixed, paraffin-embedded (FFPE) slides are scanned, and two corresponding digital images are validated by an operator. Image analysis is performed using dedicated software (Immunoscore Analyzer, HalioDx) with automated histopathological structure detection followed by operator-guided definition of tumor, healthy tissue (submucosa, muscularis propria, serosa), and epithelium (mucosa). The operator also excludes all necrotic, abscess, and artifactual (air bubble retention, dehiscence areas, background) areas to avoid false positives. The software automatically calculates the IM over 360 μm for healthy tissue and 360 μm for tumor. In the presence of multiple FFPE blocks, the one containing the IM is selected for immunoscore evaluation.

[0084] Ayers Immunity Score An additional means for predicting the antitumor efficacy of CDK4 / 6 inhibitor therapy is to determine a tumor's IFN-γ signature score and / or an extended immune signature score, as described by Ayers et al. (Ayers M, et al., "IFN-γ-Related MRNA Profile Predicts Clinical Response to PD-1 Blockade," Journal of Clinical Investigation, vol. 127, no. 8, 2017, pp. 2930-2940., doi:10.1172 / jci91190, the entire contents of which are incorporated herein by reference). They outline an exhaustive, iterative approach to construct gene expression signatures that predict response to immune checkpoint inhibitors (e.g., pembrolizumab). Starting with melanoma data, they used a one-tailed t-test to detect genes that were differentially expressed between responders and non-responders to pembrolizumab. Notably, many of these differentially expressed genes were related to IFN-γ signaling, and Ayers et al. developed a preliminary IFN-γ signature for ICI response based on average expression within the IFN-γ pathway and its associated genes.

[0085] The robustness of this preliminary response signature was further evaluated in melanoma, HNSCC, and gastric cancer. The preliminary signature showed significant associations with BOR and PFS, but not OS. To improve predictions in non-melanoma cancers, these immune signatures were trimmed and expanded by evaluating the univariate associations between individual genes and BOR and PFS within the expansion cohort. Genes with statistically insignificant associations were removed from the signature, and an intermediate IFN-gamma signature was formed in addition to those with significant associations.

[0086] As proof of concept for the expansion of such predictive signatures to multiple cancer types, we referenced the success of the preliminary and intermediate IFN-gamma signature in discriminating between responders and non-responders to anti-PD-1 therapy and developed a final signature for multiple cancer types using wide-array samples from the KEYNOTE-012 (ClinicalTrials.gov ID: NCT01848834) and KENYNOTE-028 clinical trials (ClinicalTrials.gov ID: NCT02054806). Using penalized logistic regression for BOR, we further refined the intermediate signature to a final set of 18 PD-1 / PD-L1 response-associated genes.

[0087] IFN-γ signature analysis consisted of determining the expression profiles of six genes: IDO1, CXCL10, CXCL9, HLA-DRA; STAT1, and IFN-γ. Extended immune signature analysis consisted of determining the expression profiles of 18 genes: CCL5, CD27, CD274, CD276, CD8A, CMKLR1, CXCL9, CXCR6, HLA-DRB1, HLA-DQA1, HLA-E, IDO1, LAG3, NKG7, PDCD1LG2, PSMB10, STAT1, and TIGIT.

[0088] Ayers et al. performed sequencing quantification using a 680-gene panel on the Nanostring platform. To calculate sample scores for either multigene signature (IFN-γ signature or extended immune signature), log10 transformation and subsequent quantile normalization were performed before averaging the gene set. The area under the receiver operating characteristic curve (ROC) was used as an indicator of the discriminatory power of the signature score. The Youden index (see Youden WJ. Index for rating diagnostic tests. Cancer. 1950;3(1):32-35, incorporated herein by reference), a summary measure of the ROC curve, was used as a cross-sectional method for selecting an "optimal" cutoff, i.e., a "high" or "low" signature score, to indicate potential clinical utility. For example, a "high" IFN-γ signature or extended immune signature can be determined based on a comparison with the scores of known immunogenic samples. In some embodiments, a "high" IFN-γ signature or extended immune signature score is one that is at least greater than 2.25, 2.5, or 2.75. In some embodiments, a "high" IFN-γ signature or extended immune signature score is one that is at least greater than 2.5.

[0089] This assessment provides tumor-type-independent applicability of T cell inflammatory gene expression profiles that capture the biology of the T cell inflammatory microenvironment, and as shown in the examples below, TNBC patients with high IFN-γ signature scores and / or increased immune signature scores who receive CDK4 / 6 inhibitors exhibit statistically significant improved overall survival compared to TNBC patients who do not receive CDK4 / 6 inhibitors.

[0090] The six-class immune signature of Thorsson et al. Another measure for predicting the antitumor efficacy of CDK4 / 6 inhibitor therapy is to determine a six-class immune signature of a tumor, as described by Thorsson et al., "The Immune Landscape of Cancer," Immunity, vol. 51, no. 2, 2018, pp. 812-830 (the entire contents of which are incorporated herein by reference). Thorsson et al. conducted an extensive literature search for expression signatures characterizing various phases of the immune response. This search resulted in 160 signatures for consideration. Using weighted gene co-expression network analysis (WGCNA) across the entire TCGA (The Cancer Genome Atlas) dataset, these 160 signatures were clustered into nine distinct signature modules, or signature collections, that purport to measure consistent immune phenomena. Candidate signatures were then narrowed down to the signatures from each of these nine modules that most closely resembled the "average profile."

[0091] Further examination confirmed that four of these nine representative signatures were not robust classifiers for the TCGA data, resulting in highly variable classification depending on which samples were used in model training. This left five immune signatures for use in sample classification, which are shown in Table 1 below.

[0092] [Table 1]

[0093] Scores for each of these five signatures were calculated using single-sample gene set enrichment analysis (ssGSEA) on the TCGA dataset. These data were then clustered using an unsupervised consensus clustering approach, yielding six distinct immune response subtypes, as described in Table 2.

[0094] [Table 2]

[0095] As described in Example 3, TNBC patients with tumors classified as C2 "IFN-γ dominant" who are administered a CDK4 / 6 inhibitor show a statistically significant improvement in overall survival compared to C2 "IFN-γ dominant" patients who do not receive a CDK4 / 6 inhibitor during treatment.

[0096] PD-L1 status Another means of predicting the antitumor efficacy of CDK4 / 6 inhibitor therapy is to determine the status of tumor programmed cell death-1 ligand (PD-L1). PD-L1 is a transmembrane protein that downregulates immune responses through binding to two inhibitory receptors, programmed cell death-1 (PD-1) and B7.1. PD-1 is an inhibitory receptor expressed on T cells after T cell activation and 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): pp. 739-745). Binding of PD-L1 to PD-1 inhibits T cell proliferation, cytokine production, and cytolytic activity, resulting in functional inactivation or exhaustion of T cells. 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 found on immune cells and tumor cells.See Dong H, Zhu G, Tamada K, Chen L. B7-H1, a third member of the B7 family, costimulates 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. Abnormal expression of PD-L1 on tumor cells has been reported to impede antitumor immunity and lead to immune evasion. Therefore, disruption of the PD-L1 / PD-1 pathway is an attractive strategy to reactivate tumor-specific T cell immunity suppressed by PD-L1 expression in the tumor microenvironment. In cancer, upregulation of PD-L1 can enable cancer to evade the host immune system.

[0097] PD-L1 expression can be determined by methods known in the art. For example, PD-L1 expression can be detected using PD-L1 IHC 22C3 pharmDx, an in vitro diagnostic immunohistochemistry (IHC) test developed by Dako and Bristol-Meyers Squibb and approved by the U.S. Food and Drug Administration as a companion test for pembrolizumab treatment. This is a qualitative assay using a monoclonal mouse anti-PD-L1, clone 22C3 PD-L1, and the EnVision FLEX visualization system on an Autostainer Lin 48 to detect PD-L1 in formalin-fixed, paraffin-embedded (FFPE) human non-small cell lung cancer tissue. Expression levels can be measured using the tumor proportion score (TPS), which measures the percentage of viable tumor cells that show partial or complete membrane staining. Staining can indicate PD-L1 expression between 1% and 100%.

[0098] PD-L1 expression can also be detected using PD-L1 IHC 28-8 pharmDx, an in vitro diagnostic immunohistochemistry (IHC) test developed by Dako and Merck and approved by the U.S. Food and Drug Administration as a companion test for nivolumab treatment. This qualitative assay uses a monoclonal rabbit anti-PD-L1, clone 28-8, and an En Vision FLEX visualization system on an Autostainer Lin 48 to detect PD-L1 in formalin-fixed, paraffin-embedded (FFPE) human non-small cell lung cancer tissue.

[0099] Other commercially available tests for detecting PD-L1 include the Ventana SP263 assay (developed by Ventana in collaboration with AstraZeneca) using a monoclonal rabbit anti-PD-L1 clone SP263 and the Ventana SP142 assay (developed by Ventana in collaboration with Genentech / Roche) using a rabbit monoclonal anti-PD-L1 clone SP142. Determining PD-L1 status is indication-specific and is based on either the percentage of tumor area occupied by tumor-infiltrating immune cells expressing PD-L1 (%IC) regardless of intensity or the percentage of tumor cells expressing PD-L1 (%TC) regardless of intensity. For example, in urothelial carcinoma tissue, PD-L1 expression of ≥ 5%IC is determined, for example, by the Ventana PD-L1 (SP142) assay, while PD-L1 positivity in TNBC is considered ≥ 1%IC, and in NSCLC, ≥ 50%TC or ≥ 10%IC.

[0100] Short-acting CDK4 / 6 inhibitor Short-acting CDK4 / 6 inhibitors for use in the present invention include Compound I, Compound II, Compound III, Compound IV, and Compound V, or a pharmaceutically acceptable salt thereof.

[0101] Compound I, known as trilaciclib (2'-((5-(4-methylpiperazin-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 a highly selective CDK4 / 6 inhibitor having the following structure: [ka]

[0102] As provided herein, trilaciclib, or a pharmaceutically acceptable salt thereof, composition thereof, isotopic analog, or prodrug thereof, is administered with a suitable carrier in a chemotherapy agent that induces an immune response, including chemotherapy such as an ICD-inducing chemotherapy agent. Trilaciclib is described in U.S. Patent No. 8,598,186, the entire contents of which are incorporated herein by reference. Trilaciclib can be synthesized as described in WO2019 / 0135820, the entire contents of which are incorporated herein by reference.

[0103] Trilaciclib, in one embodiment, can be administered parenterally, e.g., intravenously, to a patient prior to administration of an immune response-inducing chemotherapy, such as ICD-guided chemotherapy. In some embodiments, trilaciclib is administered within a maximum of about 24 hours, or within a maximum of about 20, 15, 10, 5, or 4 hours, e.g., within about 30-60 minutes, prior to administration of chemotherapy. In some embodiments, trilaciclib is administered approximately 22-26 hours prior to administration of chemotherapy, and again within about 4 hours, e.g., within about 30-60 minutes, prior to administration of chemotherapy. In some embodiments, the dose of trilaciclib administered is about 180 to about 280 mg / m 2 For example, dosages may be up to about 100, 125, 150, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, or 280 mg / m, as determined by a healthcare professional. 2, or any dose between these values. In certain embodiments, the dose is about 240 mg / m 2 is.

[0104] Trilaciclib can be administered in any manner that achieves the desired outcome, including systemically, parenterally, intravenously, intramuscularly, subcutaneously, or intradermally. For injection, trilaciclib can be provided, for example, as a 300 mg / vial sterile lyophilized yellow mass providing 300 mg of trilaciclib (equivalent to 349 mg of trilaciclib dihydrochloride). This product can be supplied, for example, in a single-use 20 mL clear glass vial and does not contain preservatives. Prior to administration, 300 mg / vial of trilaciclib for injection can be reconstituted with 19.5 mL of 0.9% sodium chloride injection or 5% dextrose injection. The reconstituted solution has a trilaciclib concentration of 15 mg / mL and is then generally diluted prior to intravenous administration.

[0105] In another embodiment, compound III, known as relociclib, or a pharmaceutically acceptable salt thereof, is administered instead of trilaciclib. Relociclib (2'-((5-(4-isopropylpiperazin-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) has the following chemical structure: [ka]

[0106] Lelociclib can be administered in any manner that achieves the desired effect, including systemically, parenterally, orally, intravenously, intramuscularly, subcutaneously, or intradermally. Lelociclib can be prepared as previously described in WO2014 / 144325, which is incorporated herein by reference. In some embodiments, lelociclib is administered using the same recommended amounts and methods as those described above for trilaciclib.

[0107] In yet another embodiment, a CDK4 / 6 inhibitor having the structure: [ka] or a pharmaceutically acceptable salt thereof is administered in place of trilaciclib. In some embodiments, the compound is administered using the same recommended amounts and methods as described above for trilaciclib.

[0108] In yet another embodiment, a CDK4 / 6 inhibitor having the structure: [ka] or a pharmaceutically acceptable salt thereof is administered in place of trilaciclib. In some embodiments, the compound is administered using the same recommended amounts and methods as described above for trilaciclib.

[0109] In yet another embodiment, a CDK4 / 6 inhibitor having the structure: [ka] [In the formula, R is C(H)X, NX, C(H)Y, or C(X)2; X is a linear, branched, or cyclic C1-C5 alkyl group, such as methyl, ethyl, propyl, cyclopropyl, isopropyl, butyl, sec-butyl, tert-butyl, isobutyl, cyclobutyl, pentyl, isopentyl, neopentyl, tert-pentyl, sec-pentyl, and cyclopentyl; Y is NR1R2, R1 and R2 are independently X, or R1 and R2 together are an alkyl group that forms a bridge containing one or two heteroatoms (N, O, or S); Two X groups may together form an alkyl bridge or a bridge containing one or two heteroatoms (N, S, or O) to form a spiro compound. or a pharmaceutically acceptable salt thereof is administered in place of trilaciclib. In some embodiments, a compound selected from this formula is administered using the same recommended amounts and methods as described above for trilaciclib.

[0110] In another embodiment, CDK4 / 6 inhibitors other than those specifically listed above can be used in the present invention. Non-limiting examples include palbociclib, abemaciclib, and ribociclib.

[0111] Alternatively, the CDK4 / 6 inhibitor may be formulated in any pharmaceutically effective form, such as a pill, injection or infusion solution, capsule, tablet, syrup, transdermal patch, subcutaneous patch, subcutaneous injection, dry powder, buccal or sublingual formulation, parenteral formulation, or other suitable administration formulation.

[0112] Chemotherapy that can induce immune-mediated responses Standard cancer chemotherapy can enhance tumor immunity in two major ways: (i) inducing immunogenic cell death as part of its intended therapeutic effect; and (ii) interfering with strategies used by tumors to evade immune responses. The majority of data indicates that some chemotherapeutic agents, at their standard doses and schedules, mediate their antitumor effects, at least in part, by inducing immunogenic cell death (see, e.g., Emens et al., Chemotherapy: friend of foe to cancer vaccines? Curr Opin Mol Ther 2001;3:77-84; Vanmeerbeek et al., Trial Watch: Chemotherapy-Induced Immunogenic Cell Death in Immuni-Oncology. Oncoimmunology Vol. 9, No. 1 2020:e1703449, both of which are incorporated herein by reference).

[0113] Immunogenic cell death (ICD) is a type of cell death characterized by, for example, cell surface translocation of calreticulin (CRT), extracellular release of ATP and high-mobility group box 1 (HMBG1), and stimulation of type I interferon (IFN) responses. ICD in cancer cells can prime anti-cancer immune responses. Various chemotherapeutic agents can induce ICD, as indicated by changes in the abundance and composition of tumor-infiltrating lymphocytes (TILs).

[0114] In response to ICD-inducing chemotherapeutic drugs, tumor cells express CRT on their cell surface prior to death. , Da Image-associated molecular pattern (DAMP) analysis For example, during apoptosis, ATP or release HMGB1 during secondary necrosis. These DAMPs stimulate dendritic cell (DC) recruitment to the tumor bed, tumor antigen uptake and processing, and optimal antigen presentation to T cells. Cross-priming of CD8+ T cells is induced by mature DCs and γδ T cells in an IL-1β- and IL-17-dependent manner. Primed CTLs then mount a direct cytotoxic response, killing remaining tumor cells through the production of IFN-γ, perforin-1, and granzyme B.

[0115] ICD-guided chemotherapy for use in the present invention includes alkylating agents such as cyclophosphamide, trabectedin, temozolomide, melphalan, dacarbazine, and oxaliplatin; antimetabolites such as methotrexate, mitroxantrone, gemcitabine, and 5-fluorouracil (5-FU); cytotoxic antibiotics such as bleomycin and anthracyclines (including doxorubicin, daunorubicin, epirubicin, idarubicin, and valrubicin); taxanes such as paclitaxel, cabazitaxel, and docetaxel; topoisomerase inhibitors such as topotecan, irinotecan, and etoposide; platinum compounds such as carboplatin and cisplatin; and anti-microtubule vinca alkaloid drugs such as vinblastine, vincristine, vinorelbine, and vindesine. Other ICD-inducing chemotherapy agents include bortezomib, 26S proteasome subunit inhibitors, mechlorethamine, diaziquone, mitomycin C, fludarabine, and cytosine arabinoside. In some embodiments, the ICD-inducing chemotherapy agent is selected from idarubicin, epirubicin, doxorubicin, mitoxantrone, oxaliplatin, bortezomib, gemcitabine, and cyclophosphamide, and combinations thereof. In another embodiment, the administered chemotherapeutic agent capable of inducing an immune response can modulate tumor immunity through mechanisms other than immunogenic cell death. Various chemotherapeutic agents can modulate the activity or immunophenotype of prominent immune cell subsets in tumor cells through enhanced antigen presentation, increased expression of costimulatory molecules including B7.1 (CD80) and B7.2 (CD86), downregulation of checkpoint molecules such as programmed cell death-ligand 1 (PD-L1), or promotion of tumor cell death via the Fas, perforin, or granzyme B pathways.Chemotherapy modulating tumor immunity includes drugs that eliminate myeloid-derived suppressor cell (MDSC) activity, such as gemcitabine, 5-fluorouracil, cisplatin, and doxorubicin; drugs that eliminate Treg activity, such as cyclophosphamide and 5-fluorouracil; drugs that enhance T cell cross-priming, such as paclitaxel, cisplatin, and fludarabine; and drugs that enhance T cell cross-priming, such as gemcitabine and anthracyclines (e.g., doxorubicin, daunorubicin, epirubicin, valrubicin, and idarubicin). This can be achieved by: enhancing dendritic cell activation, such as anthracyclines, taxanes, cyclophosphamide, vinca alkaloids, methotrexate, and mitomycin C; promoting an anti-tumor CD4+ T cell phenotype, such as cyclophosphamide and paclitaxel; and promoting tumor cell recognition and lysis, such as cyclophosphamide, 5-fluorouracil, paclitaxel, doxorubicin, cisplatin, and cytosine arabinoside.

[0116] In some embodiments, a method for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immune modulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor and an effective amount of an alkylating agent such as cyclophosphamide, trabectedin, temozolomide, melphalan, dacarbazine, or oxaliplatin; methotrexate, mitroxantrone, gemcitabine, or 5-fluroxantrone. The present invention provides methods comprising administering in combination with an antimetabolite such as olauracil (5-FU); a cytotoxic antibiotic such as bleomycin or an anthracycline (e.g., doxorubicin, daunorubicin, epirubicin, idarubicin, or valrubicin); a taxane such as paclitaxel, cabazitaxel, and docetaxel; a topoisomerase inhibitor such as topotecan, irinotecan, and etoposide; a platinum compound such as carboplatin and cisplatin; an anti-microtubule vinca alkaloid drug such as vinblastine, vincristine, vinorelbine, and vindesine; bortezomib; mechlorethamine; diaziconazole; fludarabine; mitomycin C; and cytosine arabinoside. In some embodiments, the administration of a CDK4 / 6 inhibitor in combination with chemotherapy does not include the administration of an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is eliminative. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as a high "IFN-γ signature" or a high "extended immune signature."In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0117] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of cyclophosphamide. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with cyclophosphamide does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is immunorejective. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0118] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising: determining whether the cancer has an immunomodulatory-favorable microenvironment, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic; and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of trabectedin. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with trabectedin does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0119] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of temozolomide. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with temozolomide does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0120] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of melphalan. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with melphalan does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0121] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of dacarbazine. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with dacarbazine does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is immunorejective. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0122] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of oxaliplatin. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with oxaliplatin does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0123] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of methotrexate. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with methotrexate does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0124] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of 5-fluorouracil (5-FU). In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with fluorouracil (5-FU) does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is immunorepressive. In some embodiments, the patient has an immunologically altered tumor that is immunorejective. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as a high "IFN-γ signature" or a high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0125] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of gemcitabine. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with gemcitabine does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0126] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of mitoxantrone. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with mitoxantrone does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0127] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of doxorubicin. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with doxorubicin does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is immunorejective. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0128] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of daunorubicin. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with daunorubicin does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0129] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of idarubicin. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with idarubicin does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0130] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of valrubicin. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with valrubicin does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0131] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of epirubicin. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with epirubicin does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0132] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of bleomycin. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with bleomycin does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor to be immunosuppressive. In some embodiments, the patient has an immunologically altered tumor to be immunoeliminating. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as a high "IFN-γ signature" or a high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0133] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of bortezomib. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with bortezomib does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0134] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of paclitaxel. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with paclitaxel does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0135] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of docetaxel. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with docetaxel does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0136] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of cabazitaxel. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with cabazitaxel does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0137] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of topotecan. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with topotecan does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0138] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of etoposide. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with etoposide does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0139] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of irinotecan. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with irinotecan does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0140] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of cisplatin. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with cisplatin does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is immunorejective. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0141] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of carboplatin. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with carboplatin does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0142] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of vinblastine. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with vinblastine does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0143] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of vincristine. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with vincristine does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0144] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of vinorelbine. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with vinorelbine does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is immunorejective. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0145] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to enhance progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of vindesine. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with vindesine does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0146] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of diaziconazole. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with diaziconazole does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0147] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of mechlorethamine. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with mechlorethamine does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0148] In some embodiments, methods are provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to enhance progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of mitomycin C. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with mitomycin C does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0149] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising the steps of: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of fludarabine. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with fludarabine does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is negligible. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as high "IFN-γ signature" or high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0150] In some embodiments, a method is provided for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to prolong progression-free survival or overall survival, the method comprising: determining whether the cancer has a surrounding microenvironment favorable to immunomodulation, whether it is immunogenically susceptible to CDK4 / 6 inhibitor treatment, or whether it is immunogenic, and, if so, administering to the patient an effective amount of a CDK4 / 6 inhibitor in combination with an effective amount of cytosine arabinoside. In some embodiments, the administration of a short-acting CDK4 / 6 inhibitor in combination with cytosine arobinoside does not include administering an immune checkpoint inhibitor. In some embodiments, the patient has a tumor classified as immunogenic. In some embodiments, the patient has an immunologically hot tumor. In some embodiments, the patient has an immunologically altered tumor that is immunosuppressive. In some embodiments, the patient has an immunologically altered tumor that is immunorepressive. In some embodiments, the patient has an immunologically altered tumor that is immunorejective. In some embodiments, the patient has a cold tumor. In some embodiments, the patient has a tumor classified as a C2 "IFN-γ dominant" class cancer. In some embodiments, the patient has a tumor classified as a high "IFN-γ signature" or a high "extended immune signature." In some embodiments, the patient has a PD-L1 positive tumor. In some embodiments, the CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof. In some embodiments, the CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0151] In any of the above embodiments, the patient to be treated is determined to have a cancer that has a surrounding microenvironment favorable to immunomodulation, is immunogenic, or is immunogenic and susceptible to CDK4 / 6 inhibitor treatment. Thus, as long as the cancer fits into the classification as described herein, the patient may be suitable for the described treatment. In some embodiments, the cancer to be treated is selected from the group consisting of breast cancer, including but not limited to estrogen receptor (ER)-positive breast cancer and triple-negative breast cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, classical Hodgkin's lymphoma (cHL), diffuse large B-cell lymphoma, bladder cancer, primary mediastinal B-cell lymphoma (PBMCL), urothelial carcinoma, microsatellite instability-high (MSI-H) solid tumors, mismatch repair-deficient (dMMR) solid tumors, gastric or gastroesophageal junction (GEJ) adenocarcinoma, esophageal squamous cell carcinoma, cervical cancer, endometrial cancer, cholangiocarcinoma, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, ovarian cancer, anal canal cancer, colorectal cancer, cutaneous melanoma, and melanoma.

[0152] In some embodiments, the present invention provides a method for selecting a patient or patient population for triple-negative breast cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to increase progression-free survival or overall survival, the method comprising the steps of administering gemcitabine and carboplatin to the patient, and administering a CDK4 / 6 inhibitor selected from Compound I, Compound II, Compound III, Compound IV, or a pharmaceutically acceptable salt thereof, prior to administration of the gemcitabine and carboplatin, wherein the cancer is determined to be immunogenic, immunogenically susceptible to CDK4 / 6 inhibitor treatment, or have a surrounding microenvironment favorable for immunomodulation prior to initiation of treatment.

[0153] In some embodiments, a method is provided for selecting a patient or patient population for triple-negative breast cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy to enhance progression-free survival or overall survival, the method comprising the steps of administering gemcitabine and carboplatin to the patient on days 1 and 8 of a 21-day breast cancer cycle, and administering a CDK4 / 6 inhibitor selected from Compound I, Compound II, Compound III, Compound IV, or a pharmaceutically acceptable salt thereof, prior to administration of the gemcitabine and carboplatin, wherein the CDK4 / 6 inhibitor is determined to be immunogenic, immunogenically susceptible to CDK4 / 6 inhibitor treatment, or have a surrounding microenvironment favorable for immunomodulation prior to initiation of treatment.

[0154] Administration Protocol The methods described herein provide for the administration of a CDK4 / 6 inhibitor and chemotherapy capable of inducing an immune-mediated response in cancer, e.g., ICD-inducing chemotherapy, to extend the overall survival or progression-free survival of cancer patients, and include the following steps: determining whether the patient can be classified as immunogenic, has a surrounding microenvironment favorable for immune modulation, or has a cancer that is susceptible to CDK4 / 6 inhibitor treatment, or whether the cancer is immunogenic and susceptible to CDK4 / 6 inhibitor treatment, and if so, administering to the patient a chemotherapeutic agent capable of inducing an immune-mediated response in combination with a CDK4 / 6 inhibitor.

[0155] In some embodiments, the CDK4 / 6 inhibitor is administered before or simultaneously with the administration of chemotherapy.In some embodiments, the selective CDK4 / 6 inhibitor is administered to the subject within about 24 hours, about 20 hours, about 16 hours, about 12 hours, about 8 hours, about 4 hours, about 2.5 hours, about 2 hours, about 1 hour, about 1 / 2 hour or less before the treatment with chemotherapy.In certain embodiments, the selective CDK4 / 6 inhibitor is administered about 1 / 2 hour before the administration of chemotherapy.

[0156] Generally, the selective CDK4 / 6 inhibitor is administered to the subject prior to chemotherapy treatment so that the CDK4 / 6 inhibitor reaches peak serum levels before or during chemotherapy treatment, allowing it to inhibit the proliferation of immune effector cells, thus protecting them from the adverse effects of chemotherapy. In some embodiments, the CDK4 / 6 inhibitor is administered simultaneously with or adjacent to chemotherapy exposure. In one embodiment, the selective CDK4 / 6 inhibitor is Compound I, or a pharmaceutically acceptable salt thereof. In one embodiment, the selective CDK4 / 6 inhibitor is Compound III, or a pharmaceutically acceptable salt thereof.

[0157] In some embodiments, the CDK4 / 6 inhibitor is administered to the subject within about 24 hours, about 20 hours, about 16 hours, about 12 hours, about 8 hours, about 4 hours, about 2.5 hours, about 2 hours, about 1 hour, about ½ hour, or less before chemotherapy. In certain embodiments, the selective CDK4 / 6 inhibitor is administered about ½ hour before chemotherapy. Generally, the selective CDK4 / 6 inhibitor is administered to the subject before chemotherapy so that the CDK4 / 6 inhibitor reaches peak serum levels before or during chemotherapy, allowing it to inhibit the proliferation of immune effector cells, thereby protecting them from the adverse effects of chemotherapy. In one embodiment, the CDK4 / 6 inhibitor is administered simultaneously with or adjacent to chemotherapy exposure. Alternatively, the CDK4 / 6 inhibitor described herein can be administered after chemotherapy exposure, if desired, to mitigate immune effector cell damage associated with chemotherapy exposure.

[0158] In some embodiments, the CDK4 / 6 inhibitor is administered twice to the subject prior to the administration of chemotherapy. For example, in some embodiments, the CDK4 / 6 inhibitor is administered approximately 18 to 28 hours prior to the administration of chemotherapy. administered to and then another dose about 4 hours, about 2.5 hours, about 2 hours, about 1 hour, about 1 / 2 hour, or less, before treatment with the chemotherapy agent. In certain embodiments, the selective CDK4 / 6 inhibitor is administered about 22 to 26 hours before administration of the chemotherapy agent. is administered to It is re-administered approximately 1 / 2 hour before the chemotherapy dose.

[0159] In certain embodiments, the CDK4 / 6 inhibitor is administered prior to or concurrently with the administration of a chemotherapy agent, for example, on days 1-3 every 21 days; on days 1-3 every 28 days; on day 1 every 3 weeks; on days 1, 8, and 15 every 28 days, on days 1 and 8 every 28 days; on days 1 and 8 every 21 days; on days 1-5 every 21 days; on day 1 of the week for 6-8 weeks; on days 1, 22, and 43; on days 1 and 2 every week; on days 1-4 and 22-25; on days 1-4; on days 22-25 and 43-46; and similar types of chemotherapy regimens. In some embodiments, the CDK4 / 6 inhibitor is administered prior to or concurrently with at least one dose of the chemotherapy agent during the chemotherapy regimen. In some embodiments, the CDK4 / 6 inhibitor is administered prior to or concurrently with one or more administrations of the chemotherapeutic agent during a chemotherapeutic drug administration regimen. In one embodiment, the CDK4 / 6 inhibitor is administered prior to or concurrently with each administration of the chemotherapeutic agent during a chemotherapeutic drug administration regimen.

[0160] In some embodiments, the CDK4 / 6 inhibitor is administered during a standard chemotherapy protocol, such as a 21-day cycle, for example, before or simultaneously with each chemotherapy administration. After the standard chemotherapy protocol is stopped, the CDK4 / 6 inhibitor is further administered alone at a maintenance dose. In some embodiments, the CDK4 / 6 inhibitor is further administered once per week for at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 26, 52, 104 weeks, or longer. In some embodiments, the CDK4 / 6 inhibitor is administered once every 21 days after the chemotherapy protocol is stopped. In one embodiment, the selective CDK4 / 6 inhibitor is a fast-acting, short-half-life CDK4 / 6 inhibitor.

[0161] In some embodiments, the CDK4 / 6 inhibitor is administered in conjunction with a chemotherapy agent in a maintenance dosing regimen after cessation of a standard chemotherapy protocol. Maintenance therapy can comprise either continuation of a given agent as part of a first-line or previous regimen (continuation maintenance) or treatment with a new agent (switch maintenance).

[0162] In some embodiments, the CDK4 / 6 inhibitor is further administered in a maintenance treatment regimen, in which the CDK4 / 6 inhibitor is administered in combination with a reduced maintenance dose of chemotherapy at a specified administration interval, for example, but not limited to, once a week, once every two weeks, once every three weeks, once a month, once every six weeks, once every two months, once every three months, or once every six months after the end of the first chemotherapy treatment. In some embodiments, the CDK4 / 6 inhibitor is administered together with the same agent as that used in the previous chemotherapy treatment phase. In some embodiments, the CDK4 / 6 inhibitor is administered together with a different chemotherapeutic agent than that used in the previous chemotherapy treatment phase.

[0163] In some embodiments, the patient is not administered a checkpoint inhibitor.

[0164] Embodiment The following embodiments are provided herein:

[0165] Embodiment 1 1. A method for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy, to enhance progression-free survival or overall survival, comprising: The method comprises the steps of: (i) determining whether the patient's cancer has a surrounding microenvironment that is favorable for immune modulation; (ii) determining whether the chemotherapy regimen induces an immune-mediated response, e.g., immunogenic cell death; If both (i) and (ii) are affirmative, then: (iii) Compound I, II, III, IV, or V: [ka] [In the formula, R is C(H)X, NX, C(H)Y, or C(X)2; X is a linear, branched, or cyclic C1-C5 alkyl group, such as methyl, ethyl, propyl, cyclopropyl, isopropyl, butyl, sec-butyl, tert-butyl, isobutyl, cyclobutyl, pentyl, isopentyl, neopentyl, tert-pentyl, sec-pentyl, and cyclopentyl; Y is NR1R2, R1 and R2 are independently X, or R1 and R2 together are an alkyl group that forms a bridge containing one or two heteroatoms (N, O, or S); Two X groups may together form an alkyl bridge or a bridge containing one or two heteroatoms (N, S, or O) to form a spiro compound. or a pharmaceutically acceptable salt thereof. comprising said CDK4 / 6 inhibitor is administered prior to, or optionally prior to and concurrently with, the administration of said chemotherapy; The method, wherein the prolongation of progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone.

[0166] Embodiment 2 2. The method of embodiment 1, wherein determining whether the cancer has a surrounding microenvironment that is favorable for immune modulation comprises comparing the cancer tissue sample with the cancer tissue sample characterized in FIG.

[0167] Embodiment 3 2. The method of embodiment 1, wherein determining whether said cancer has a surrounding microenvironment that is favorable for immunomodulation comprises evaluating said cancer according to FIG.

[0168] Embodiment 4 2. The method of embodiment 1, wherein determining whether the cancer has a surrounding microenvironment favorable to immunomodulation is according to the Galon immunoscore system.

[0169] Embodiment 5 2. The method of embodiment 1, wherein determining whether the cancer has a surrounding microenvironment that is favorable for immunomodulation comprises assessing whether the cancer has sufficiently high levels of major histocompatibility complex class I antigens available to elicit an effective immune response.

[0170] Embodiment 6 2. The method of embodiment 1, wherein determining whether the cancer has a surrounding microenvironment that is favorable for immunomodulation comprises assessing whether the cancer has sufficiently high levels of major histocompatibility complex class II antigens available to elicit an effective immune response.

[0171] Embodiment 7 2. The method of embodiment 1, wherein determining whether the cancer has a surrounding microenvironment that is favorable for immunomodulation comprises assessing whether the cancer has sufficiently high levels of major histocompatibility complex class I and class II antigens available to elicit an effective immune response.

[0172] Embodiment 8 The method of any of embodiments 1-7, wherein said patient has a cancer that is immunogenically classified as an immunologically hot tumor.

[0173] Embodiment 9 The method of any of embodiments 1-7, wherein said patient has a cancer that is immunogenically classified as an immunosuppressively altered tumor.

[0174] Embodiment 10 8. The method of any of embodiments 1-7, wherein determining whether the cancer has a surrounding microenvironment that is favorable for immune modulation comprises assessing whether the cancer is of the IFN-γ dominant class of cancer, whether it has a cancer microenvironment with a high IFN-γ signature, or whether it has a high extended immune signature, PD-L1 positivity, or a combination thereof.

[0175] Embodiment 11 The method of any of embodiments 1-7, wherein said CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof.

[0176] Embodiment 12 The method of any of embodiments 1-11, wherein said CDK4 / 6 inhibitor administered is Compound II, or a pharmaceutically acceptable salt thereof.

[0177] Embodiment 13 The method of any of embodiments 1-11, wherein said CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0178] Embodiment 14 The method of any of embodiments 1-11, wherein said CDK4 / 6 inhibitor administered is Compound IV, or a pharmaceutically acceptable salt thereof.

[0179] Embodiment 15 The method of any of embodiments 1-11, wherein said CDK4 / 6 inhibitor administered is Compound V, or a pharmaceutically acceptable salt thereof.

[0180] Embodiment 16 16. The method of any of embodiments 1-15, wherein said CDK4 / 6 inhibitor is administered within about 24 hours prior to administration of said chemotherapy.

[0181] Embodiment 17 16. The method of any of embodiments 1-15, wherein said CDK4 / 6 inhibitor is administered within about 4 hours prior to administration of said chemotherapy.

[0182] Embodiment 18 16. The method of any of embodiments 1-15, wherein said CDK4 / 6 inhibitor is administered within about 30 minutes prior to administration of said chemotherapy.

[0183] Embodiment 19 19. The method of any of embodiments 1-18, wherein said chemotherapy is selected from the group consisting of cyclophosphamide, trabectedin, temozolomide, melphalan, dacarbazine, oxaliplatin, methotrexate, mitroxatrone, gemcitabine, 5-fluorouracil (5-FU), bleomycin, doxorubicin, daunorubicin, epirubicin, idarubicin, valrubicin, paclitaxel, cabazitaxel, docetaxel, topotecan, irinotecan, etoposide, carboplatin, cisplatin, bortezomib, vinblastine, vincristine, vindesine, vinorelbine, diaziquone, mechlorethamine, mitomycin C, fludarabine, cytosine arabinoside, and combinations thereof.

[0184] Embodiment 20 1. A method for selecting a patient or patient population for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy, to enhance progression-free survival or overall survival, comprising: The method comprises the steps of: (i) determining whether the cancer is immunogenic; (ii) determining whether the patient is amenable to the cancer-based ICD-guided chemotherapy; (iii) when it is determined that the cancer is immunogenic and that the ICD-guided chemotherapy is administrable, administering an effective amount of the ICD-guided chemotherapy in combination with an effective amount of a short-acting CDK4 / 6 inhibitor selected from Compound I, Compound II, Compound III, Compound IV, or Compound V, or a pharmaceutically acceptable salt thereof. comprising The method, wherein the CDK4 / 6 inhibitor is administered prior to administration of the ICD-guided chemotherapy, or optionally prior to and concurrently with the ICD-guided chemotherapy.

[0185] Embodiment 21 21. The method of embodiment 20, wherein the cancer is immunogenic if the cancer has a surrounding microenvironment that is favorable to immune modulation, and determining which comprises comparing the cancer tissue sample with the cancer tissue sample characterized in FIG.

[0186] Embodiment 22 21. The method of embodiment 20, wherein the cancer is immunogenic if the cancer has a surrounding microenvironment that is favorable to immune modulation, and determining which comprises evaluating the cancer according to FIG. 6.

[0187] Embodiment 23 21. The method of embodiment 20, wherein the cancer is immunogenic if it has a surrounding microenvironment that is favorable for immune modulation according to the Galon immunoscore system.

[0188] Embodiment 24 21. The method of embodiment 20, wherein said cancer is immunogenic if said cancer is immunogenically classified as an immunologically hot tumor.

[0189] Embodiment 25 21. The method of embodiment 20, wherein said cancer is immunogenic if said cancer is immunogenically classified as an immunologically altered immunosuppressive tumor.

[0190] Embodiment 26 The method of embodiment 20, wherein said patient has a cancer that is immunogenically classified as elimination-altered.

[0191] Embodiment 27 21. The method of embodiment 20, wherein the cancer is immunogenic if it is classified as an IFN-γ dominant class of cancer, if it has a cancer microenvironment with a high IFN-γ signature, or if it has a high extended immune signature, is PD-L1 positive, or a combination thereof.

[0192] Embodiment 28 The method of any of embodiments 20-27, wherein said CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof.

[0193] Embodiment 29 The method of any of embodiments 20-27, wherein said CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0194] Embodiment 30 The method of any of embodiments 20-29, wherein said CDK4 / 6 inhibitor is administered within about 24 hours prior to administration of said ICD-inducing chemotherapy.

[0195] Embodiment 31 The method of any of embodiments 20-29, wherein said CDK4 / 6 inhibitor is administered within about 4 hours prior to administration of said ICD-inducing chemotherapy.

[0196] Embodiment 32 The method of any of embodiments 20-29, wherein said CDK4 / 6 inhibitor is administered within about 30 minutes prior to administration of said ICD-guided chemotherapy.

[0197] Embodiment 33 The method of any of embodiments 20-29, wherein the CDK4 / 6 inhibitor is first administered about 22-26 hours before administration of the ICD-guided chemotherapy and re-administered within about 4 hours before administration of the ICD-guided chemotherapy.

[0198] Embodiment 34 34. The method of any of embodiments 20-33, wherein said ICD-guided chemotherapy is selected from the group consisting of cyclophosphamide, trabectedin, temozolomide, melphalan, dacarbazine, oxaliplatin, methotrexate, mitroxatrone, gemcitabine, 5-fluorouracil (5-FU), bleomycin, doxorubicin, daunorubicin, epirubicin, idarubicin, valrubicin, paclitaxel, cabazitaxel, docetaxel, topotecan, irinotecan, etoposide, carboplatin, cisplatin, bortezomib, vinblastine, vincristine, vindesine, vinorelbine, diaziquone, mechlorethamine, mitomycin C, fludarabine, cytosine arabinoside, and combinations thereof.

[0199] Embodiment 35 The method of any of embodiments 1-34, wherein said patient is not administered an immune checkpoint inhibitor concurrently with administration of said CDK4 / 6 inhibitor.

[0200] Embodiment 36 1. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy in a patient or patient population selected for extending progression-free survival or overall survival, comprising: The patient or the selection of the patient comprises the steps of: (i) selecting the patient or patient population based on determining whether the cancer has a surrounding microenvironment that is favorable for immune modulation and determining whether the chemotherapy regimen is capable of inducing an immune-mediated response; (ii) administering said CDK4 / 6 inhibitor prior to administration of said chemotherapy, or optionally prior to and concurrently with said chemotherapy. comprising The use, wherein the improvement in overall survival or progression-free survival is compared to the overall survival or progression-free survival based on administration of the chemotherapy alone.

[0201] Embodiment 37 The use of embodiment 36, wherein determining whether the cancer has a surrounding microenvironment favorable to immunomodulation comprises comparing the cancer tissue sample with the cancer tissue sample characterized in Figure 7.

[0202] Embodiment 38 The use of embodiment 36, wherein determining whether said cancer has a surrounding microenvironment favorable to immunomodulation comprises assessing said cancer according to Figure 6.

[0203] Embodiment 39 The use of embodiment 36, wherein determining whether the cancer has a surrounding microenvironment favorable to immunomodulation is according to the Galon immunoscore system.

[0204] Embodiment 40 The use of embodiment 36, wherein determining whether the cancer has a surrounding microenvironment that is favorable for immunomodulation comprises assessing whether the cancer's microenvironment has sufficiently high levels of major histocompatibility complex class I antigens available to elicit an effective immune response.

[0205] Embodiment 41 The use of embodiment 36, wherein determining whether the cancer has a surrounding microenvironment that is favorable for immunomodulation comprises assessing whether the cancer's microenvironment has sufficiently high levels of major histocompatibility complex class II antigens available to elicit an effective immune response.

[0206] Embodiment 42 The use of embodiment 36, wherein determining whether the cancer has a surrounding microenvironment that is favorable for immunomodulation comprises assessing whether the cancer's microenvironment has sufficiently high levels of major histocompatibility complex class I and class II antigens available to elicit an effective immune response.

[0207] Embodiment 43 The use of embodiment 36, wherein said patient has a cancer that is immunogenically classified as an immunologically hot tumor.

[0208] EMBODIMENT 44 The use of embodiment 36, wherein said patient has a cancer immunogenically classified as an immunosuppressively altered tumor.

[0209] Embodiment 45 The use of embodiment 36, wherein said patient has a cancer that is a C2 IFN-γ dominant class of cancer, a cancer microenvironment with a high IFN-γ signature or an extended immune signature, or a cancer that is PD-L1 positive.

[0210] Embodiment 46 The use of embodiment 36, wherein determining whether the cancer has a surrounding microenvironment that is favorable for immunomodulation comprises assessing whether the cancer's microenvironment has a sufficiently high degree of T cell and cytotoxic T cell infiltration.

[0211] Embodiment 47 The use of any of embodiments 36-46, wherein said CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof.

[0212] Embodiment 48 The use of any of embodiments 36-46, wherein said CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0213] Embodiment 49 The use of any of embodiments 36-46, wherein said CDK4 / 6 inhibitor is administered within about 24 hours prior to administration of said chemotherapy.

[0214] Embodiment 50 The use of any of embodiments 36-49, wherein the ICD-guided chemotherapy is selected from the group consisting of cyclophosphamide, trabectedin, temozolomide, melphalan, dacarbazine, oxaliplatin, methotrexate, mitroxatrone, gemcitabine, 5-fluorouracil (5-FU), bleomycin, doxorubicin, daunorubicin, epirubicin, idarubicin, valrubicin, paclitaxel, cabazitaxel, docetaxel, topotecan, irinotecan, etoposide, carboplatin, cisplatin, bortezomib, vinblastine, vincristine, vindesine, vinorelbine, diaziquone, mechlorethamine, mitomycin C, fludarabine, cytosine arabinoside, and combinations thereof.

[0215] Embodiment 51 The use of any of embodiments 36-49, wherein said chemotherapy is immunogenic cell death (ICD)-inducing chemotherapy.

[0216] Embodiment 52 The use of any of embodiments 36-51, wherein said cancer is selected from the group consisting of triple-negative breast cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, classical Hodgkin's lymphoma (cHL), bladder cancer, primary mediastinal B-cell lymphoma (PBMCL), urothelial carcinoma, microsatellite instability-high (MSI-H) solid tumors, mismatch repair-deficient (dMMR) solid tumors, gastric or gastroesophageal junction (GEJ) adenocarcinoma, esophageal squamous cell carcinoma, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, ovarian cancer, anal canal cancer, colorectal cancer, and melanoma.

[0217] Embodiment 53 The use of any of embodiments 36-52, wherein said patient is not administered an immune checkpoint inhibitor concurrently with administration of said CDK4 / 6 inhibitor.

[0218] EMBODIMENT 54 1. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy in a patient or patient population selected for extending progression-free survival or overall survival, comprising: The patient or the selection of the patient comprises the steps of: (i) determining whether the cancer is immunogenically susceptible to CDK4 / 6 inhibitor treatment; (ii) determining whether the patient can be administered chemotherapy capable of inducing an immune-mediated response; (iii) if both (i) and (ii) are positive, administering an effective amount of said CDK4 / 6 inhibitor before administration of said chemotherapy, or optionally before and simultaneously with said chemotherapy. comprising The use, wherein the improvement in overall survival or progression-free survival is compared to the overall survival or progression-free survival based on administration of the chemotherapy alone.

[0219] Embodiment 55 The use of embodiment 54, wherein said cancer is immunogenically sensitive to said CDK4 / 6 inhibitor treatment if said cancer has a surrounding microenvironment favorable to immunomodulation as assessed according to Figure 6.

[0220] Embodiment 56 The use of embodiment 54, wherein said cancer is immunogenically sensitive to said CDK4 / 6 inhibitor treatment if said cancer has a surrounding microenvironment that is favorable for immune modulation as assessed according to the Galon Immunoscore system.

[0221] Embodiment 57 The use of embodiment 54, wherein said cancer is immunogenically susceptible to said CDK4 / 6 inhibitor treatment when the microenvironment of said cancer has sufficiently high levels of major histocompatibility complex class I antigens available to elicit an effective immune response.

[0222] Embodiment 58 The use of embodiment 54, wherein said cancer is immunogenically susceptible to said CDK4 / 6 inhibitor treatment when the microenvironment of said cancer has sufficiently high levels of major histocompatibility complex class II antigens available to elicit an effective immune response.

[0223] Embodiment 59 The use of embodiment 54, wherein said cancer is immunogenically susceptible to said CDK4 / 6 inhibitor treatment when the microenvironment of said cancer has sufficiently high levels of major histocompatibility complex class I and class II antigens available to elicit an effective immune response.

[0224] Embodiment 60 The use of any of embodiments 54-59, wherein said patient has a cancer that is immunogenically classified as an immunologically hot tumor.

[0225] Embodiment 61 The use of any of embodiments 54-59, wherein said patient has a cancer that is immunogenically classified as an immunosuppressively altered tumor.

[0226] Embodiment 62 The use of embodiment 54, wherein said patient has a cancer microenvironment that is a C2 IFN-γ dominant class of cancer, or has a cancer microenvironment with a high IFN-γ signature or a high extended immune signature, or has a cancer that is PD-L1 positive.

[0227] Embodiment 63 The use of embodiment 54, wherein said cancer is immunogenically susceptible to said CDK4 / 6 inhibitor treatment when the microenvironment of said cancer has a sufficiently high degree of T cell and cytotoxic T cell infiltration.

[0228] EMBODIMENT 64 The use of any of embodiments 54-63, wherein said CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof.

[0229] Embodiment 65 The use of any of embodiments 54-63, wherein said CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0230] Embodiment 66 The use of any of embodiments 54-65, wherein said CDK4 / 6 inhibitor is administered within about 24 hours prior to administration of said chemotherapy.

[0231] Embodiment 67 The use of any of embodiments 54-66, wherein said CDK4 / 6 inhibitor is administered within about 4 hours before administration of said chemotherapy.

[0232] Embodiment 68 The use of any of embodiments 54-66, wherein said CDK4 / 6 inhibitor is administered within about 30 minutes prior to administration of said chemotherapy.

[0233] Embodiment 69 The use of any of embodiments 54-69, wherein the ICD-guided chemotherapy is selected from the group consisting of cyclophosphamide, trabectedin, temozolomide, melphalan, dacarbazine, oxaliplatin, methotrexate, mitroxatrone, gemcitabine, 5-fluorouracil (5-FU), bleomycin, doxorubicin, daunorubicin, epirubicin, idarubicin, valrubicin, paclitaxel, cabazitaxel, docetaxel, topotecan, irinotecan, etoposide, carboplatin, cisplatin, bortezomib, vinblastine, vincristine, vindesine, vinorelbine, diaziquone, mechlorethamine, mitomycin C, fludarabine, cytosine arabinoside, and combinations thereof.

[0234] Embodiment 70 The use of any of embodiments 54-69, wherein said chemotherapy is immunogenic cell death (ICD)-inducing chemotherapy.

[0235] Embodiment 71 The use of any of embodiments 54-70, wherein said cancer is selected from the group consisting of triple-negative breast cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, classical Hodgkin's lymphoma (cHL), bladder cancer, primary mediastinal B-cell lymphoma (PBMCL), urothelial carcinoma, microsatellite instability-high (MSI-H) solid tumors, mismatch repair-deficient (dMMR) solid tumors, gastric or gastroesophageal junction (GEJ) adenocarcinoma, esophageal squamous cell carcinoma, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, ovarian cancer, anal canal cancer, colorectal cancer, and melanoma.

[0236] Embodiment 72 The use of any of embodiments 54-71, wherein said patient is not administered an immune checkpoint inhibitor concurrently with administration of said CDK4 / 6 inhibitor.

[0237] Embodiment 73 The use of any of embodiments 38-72, wherein said patient is administered about 22-26 hours before the first dose of said chemotherapy and re-administered within about 4 hours before the first dose of said chemotherapy.

[0238] EMBODIMENT 74 1. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy in a patient or patient population selected for extending progression-free survival or overall survival, comprising: The patient or the selection of the patient comprises the steps of: (i) determining whether the cancer is immunogenically susceptible to CDK4 / 6 inhibitor treatment; (ii) determining whether the patient can be administered chemotherapy that induces an immune response based on the cancer, e.g., ICD-inducing chemotherapy; (iii) if it is determined that the cancer is immunogenically susceptible to treatment with the CDK4 / 6 inhibitor and that the chemotherapy that induces an immune response is administrable, administering an effective amount of the chemotherapy in combination with an effective amount of the CDK4 / 6 inhibitor. comprising the CDK4 / 6 inhibitor is administered prior to, or prior to and concurrently with, the administration of the chemotherapy; The use, wherein the improved progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone.

[0239] Embodiment 75 The use of embodiment 74, wherein determining whether said cancer is immunogenically susceptible to said CDK4 / 6 inhibitor treatment comprises comparing a cancer tissue sample with the cancer tissue sample characterized in Figure 7.

[0240] Embodiment 76 The use of embodiment 74, wherein determining whether said cancer is immunogenically susceptible to said CDK4 / 6 inhibitor treatment comprises assessing the cancer according to Figure 6.

[0241] Embodiment 77 The use of embodiment 74, wherein determining whether said cancer is immunogenically sensitive to said CDK4 / 6 inhibitor treatment is according to the Galon immunoscore system.

[0242] Embodiment 78 The use of embodiment 74, wherein determining whether the cancer is immunogenically susceptible to the CDK4 / 6 inhibitor treatment comprises assessing whether the cancer's microenvironment has sufficiently high levels of major histocompatibility complex class I antigens available to elicit an effective immune response.

[0243] Embodiment 79 The use of embodiment 74, wherein determining whether the cancer is immunogenically susceptible to the CDK4 / 6 inhibitor treatment comprises assessing whether the cancer's microenvironment has sufficiently high levels of major histocompatibility complex class II antigens available to elicit an effective immune response.

[0244] Embodiment 80 The use of embodiment 74, wherein determining whether the cancer is immunogenically sensitive to the CDK4 / 6 inhibitor comprises assessing whether the cancer's microenvironment has sufficiently high levels of major histocompatibility complex class I and class II antigens available to elicit an effective immune response.

[0245] Embodiment 81 The use of any of embodiments 74-79, wherein said patient has a cancer that is immunogenically classified as an immunologically hot tumor.

[0246] Embodiment 82 The use of any of embodiments 74-79, wherein said patient has a cancer that is immunogenically classified as an immunosuppressively altered tumor.

[0247] Embodiment 83 The use of any of embodiments 74-79, wherein the patient has a cancer that is a C2 IFN-γ dominant class of cancer, has a cancer microenvironment with a high IFN-γ signature or a high extended immune signature, or has a cancer that is PD-L1 positive.

[0248] Embodiment 84 The use of embodiment 74, wherein determining whether the cancer is immunogenically sensitive to the CDK4 / 6 inhibitor treatment comprises assessing whether the cancer microenvironment has a sufficiently high degree of T cell and cytotoxic T cell infiltration.

[0249] Embodiment 85 The use of embodiment 74, wherein determining whether the cancer is immunogenically susceptible to the CDK4 / 6 inhibitor treatment comprises assessing whether the cancer has genomic instability and whether the microenvironment indicates the presence of a pre-existing anti-tumor immune response.

[0250] Embodiment 86 The use of any of embodiments 74-85, wherein said CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof.

[0251] Embodiment 87 The use of any of embodiments 74-85, wherein said CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0252] Embodiment 88 The use of any of embodiments 74-85, wherein said CDK4 / 6 inhibitor is administered within about 24 hours prior to administration of said chemotherapy.

[0253] Embodiment 89 The use of any of embodiments 74-85, wherein said CDK4 / 6 inhibitor is administered within about 4 hours before administration of said chemotherapy.

[0254] Embodiment 90 The use of any of embodiments 74-85, wherein said CDK4 / 6 inhibitor is administered within about 30 minutes prior to administration of said chemotherapy.

[0255] Embodiment 91 The use of any of embodiments 74-90, wherein said ICD-guided chemotherapy is selected from the group consisting of cyclophosphamide, trabectedin, temozolomide, melphalan, dacarbazine, oxaliplatin, methotrexate, mitroxatrone, gemcitabine, 5-fluorouracil (5-FU), bleomycin, doxorubicin, daunorubicin, epirubicin, idarubicin, valrubicin, paclitaxel, cabazitaxel, docetaxel, topotecan, irinotecan, etoposide, carboplatin, cisplatin, bortezomib, vinblastine, vincristine, vindesine, vinorelbine, diaziquone, mechlorethamine, mitomycin C, fludarabine, cytosine arabinoside, and combinations thereof.

[0256] Embodiment 92 The use of any of embodiments 74 to 90, wherein said chemotherapy is immunogenic cell death (ICD)-inducing chemotherapy.

[0257] Embodiment 93 The use of any of embodiments 74-92, wherein said cancer is selected from the group consisting of triple-negative breast cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, classical Hodgkin's lymphoma (cHL), bladder cancer, primary mediastinal B-cell lymphoma (PBMCL), urothelial carcinoma, microsatellite instability-high (MSI-H) solid tumors, mismatch repair-deficient (dMMR) solid tumors, gastric or gastroesophageal junction (GEJ) adenocarcinoma, esophageal squamous cell carcinoma, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, ovarian cancer, anal canal cancer, colorectal cancer, and melanoma.

[0258] Embodiment 94 The use of any of embodiments 74-93, wherein said patient is not administered an immune checkpoint inhibitor concurrently with administration of said CDK4 / 6 inhibitor.

[0259] Embodiment 95 The use of any of embodiments 74-93, wherein said patient is administered about 22-26 hours before the first administration of said chemotherapy and re-administered within about 4 hours before the first administration of said chemotherapy.

[0260] Embodiment 96 The use of any of embodiments 74-95, wherein said patient is not administered an immune checkpoint inhibitor concurrently with administration of said CDK4 / 6 inhibitor.

[0261] Embodiment 97 1. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy in a patient or patient population selected for extending progression-free survival or overall survival, comprising: The patient or the selection of the patient comprises the steps of: (i) determining whether the cancer is IFN-γ dominant; (ii) determining whether the patient can be administered chemotherapy that induces an immune response; (iii) when it is determined that the cancer is IFN-γ dominant and that the chemotherapy that induces an immune response is administrable, administering an effective amount of the chemotherapy in combination with an effective amount of a CDK4 / 6 inhibitor. comprising the CDK4 / 6 inhibitor is administered prior to, or prior to and concurrently with, the administration of the chemotherapy; The use, wherein the improved progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone.

[0262] Embodiment 98 The use of embodiment 97, wherein determining whether the cancer is IFN-γ dominant is based on a cancer microenvironment with high M1 / M2 polarization, strong CD8+ T cell staining, and high T cell receptor diversity.

[0263] Embodiment 99 The use of embodiment 97, wherein determining whether the cancer is IFN-γ dominant is based on the classification of the 6-class immune signature score of Thorsson et al.

[0264] Embodiment 100 1. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy in a patient or patient population selected for extending progression-free survival or overall survival, comprising: The patient or the selection of the patient comprises the steps of: (i) determining whether the cancer has an elevated IFN-γ signature; (ii) determining whether the patient can be administered chemotherapy that induces an immune response; (iii) when it is determined that the cancer has a high IFN-γ signature and that the chemotherapy that induces an immune response is administrable, administering an effective amount of the chemotherapy in combination with an effective amount of a CDK4 / 6 inhibitor. comprising the CDK4 / 6 inhibitor is administered prior to, or prior to and concurrently with, the administration of the chemotherapy; The use, wherein the improved progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone.

[0265] Embodiment 101 The use of embodiment 100, wherein determining whether the cancer has a high IFN-γ signature is based on the expression levels of the genes IDO1, CXCL10, CSCL9, HLA-DRA, STAT1, and IFN-γ in the tumor microenvironment.

[0266] Embodiment 102 The use of embodiment 100, wherein determining whether the cancer has a high IFN-γ signature is based on a high Ayers et al. IFN-γ signature score.

[0267] Embodiment 103 1. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy in a patient or patient population selected for extending progression-free survival or overall survival, comprising: The patient or the selection of the patient comprises the steps of: (i) determining whether the cancer has an elevated extended immunological signature; (ii) determining whether the patient can be administered chemotherapy that induces an immune response; (iii) if it is determined that the cancer has a highly expanded immunological signature and that the chemotherapy that induces an immune response is administerable, administering an effective amount of the chemotherapy in combination with an effective amount of a CDK4 / 6 inhibitor. comprising the CDK4 / 6 inhibitor is administered prior to, or prior to and concurrently with, the administration of the chemotherapy; The use, wherein the improved progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone.

[0268] Embodiment 104 The use of embodiment 103, wherein determining whether the cancer has an elevated extended immunological signature is based on the expression levels of the genes CCL5, CD27, CD274, CD276, CD8A, CMKLR1, CXCL9, CXCR6, HLA-DRB1, HLA-DQA1, HLA-E, IDO1, LAG3, NKG7, PDCD1LG2, PSMB10, STAT1, and TIGIT in the tumor microenvironment.

[0269] Embodiment 105 The use of embodiment 103, wherein determining whether the cancer has a high extended immunological signature is based on the Ayers et al. extended immune signature score.

[0270] Embodiment 106 1. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy in a patient or patient population selected for extending progression-free survival or overall survival, comprising: The patient or the selection of the patient comprises the steps of: (i) determining whether the cancer is a hot tumor; (ii) determining whether the patient is amenable to chemotherapy that induces an immune response; (iii) when it is determined that the cancer is a hot tumor and the chemotherapy that induces an immune response is administrable, administering an effective amount of the chemotherapy in combination with an effective amount of a CDK4 / 6 inhibitor. comprising the CDK4 / 6 inhibitor is administered prior to, or prior to and concurrently with, the administration of the chemotherapy; The use, wherein the improved progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone.

[0271] Embodiment 107 The use of embodiment 106, wherein determining whether the cancer is a hot tumor comprises comparing the cancer tissue sample with the cancer tissue sample characterized in Figure 7.

[0272] Embodiment 108 The use of embodiment 106, wherein determining whether the cancer is a hot tumor comprises assessing the cancer according to Figure 6.

[0273] Embodiment 109 The use of embodiment 106, wherein determining whether the cancer is a hot tumor is according to the Galon immunoscore system.

[0274] Embodiment 110 The use of embodiment 106, wherein determining whether the cancer is a hot tumor comprises assessing whether the microenvironment of the cancer has sufficiently high levels of major histocompatibility complex class I antigens available to elicit an effective immune response.

[0275] Embodiment 111 The use of embodiment 106, wherein determining whether the cancer is a hot tumor comprises assessing whether the microenvironment of the cancer has sufficiently high levels of major histocompatibility complex class II antigens available to elicit an effective immune response.

[0276] Embodiment 112 The use of embodiment 106, wherein determining whether the cancer is a hot tumor comprises assessing whether the cancer's microenvironment has sufficiently high levels of major histocompatibility complex class I and class II antigens available to elicit an effective immune response.

[0277] Embodiment 113 The use of embodiment 106, wherein determining whether the cancer is a hot tumor comprises assessing whether the microenvironment of the cancer has a sufficiently high degree of T cell and cytotoxic T cell infiltration.

[0278] Embodiment 114 The use of embodiment 106, wherein determining whether the cancer is a hot tumor comprises assessing whether the microenvironment of the cancer has immune checkpoint activation selected from programmed cell death protein 1 (PD-1) expression and cytotoxic T-lymphocyte-associated antigen 4 (CTLA4) expression.

[0279] Embodiment 115 The use of embodiment 106, wherein determining whether the cancer is a hot tumor comprises assessing whether the microenvironment of the cancer has T-cell immunoglobulin mucin receptor 3 (TIM3) expression and lymphocyte activation gene 3 (LAG3) expression.

[0280] Embodiment 116 The use of embodiment 106, wherein determining whether the cancer is a hot tumor comprises assessing whether the microenvironment of the cancer has impaired T cell function.

[0281] Embodiment 117 The use of embodiment 106, wherein determining whether the cancer is a hot tumor comprises assessing whether the cancer has genomic instability and whether the microenvironment indicates the presence of a pre-existing anti-tumor immune response.

[0282] Embodiment 118 1. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy in a patient or patient population selected for extending progression-free survival or overall survival, comprising: The patient or the selection of the patient comprises the steps of: (i) determining whether the cancer is PD-L1 positive; (ii) determining whether the patient can be administered chemotherapy that induces an immune response; (iii) when the cancer is PD-L1 positive and it is determined that the chemotherapy that induces an immune response can be administered, administering an effective amount of the chemotherapy in combination with an effective amount of a CDK4 / 6 inhibitor. comprising the CDK4 / 6 inhibitor is administered prior to, or prior to and concurrently with, the administration of the chemotherapy; The use, wherein the improved progression-free survival or overall survival is compared to the progression-free survival or overall survival based on administration of the chemotherapy alone.

[0283] Embodiment 119 The use of any of embodiments 97-118, wherein said CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof.

[0284] Embodiment 120 The use of any of embodiments 97-118, wherein said CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0285] Embodiment 121 The use of any of embodiments 97-118, wherein said CDK4 / 6 inhibitor is administered within about 24 hours prior to administration of said chemotherapy.

[0286] Embodiment 122 The use of any of embodiments 97-118, wherein said CDK4 / 6 inhibitor is administered within about 4 hours prior to administration of said chemotherapy.

[0287] Embodiment 123 The use of any of embodiments 97-118, wherein said CDK4 / 6 inhibitor is administered within about 30 minutes prior to administration of said chemotherapy.

[0288] Embodiment 124 The use of any of embodiments 97-123, wherein the ICD-guided chemotherapy is selected from the group consisting of cyclophosphamide, trabectedin, temozolomide, melphalan, dacarbazine, oxaliplatin, methotrexate, mitroxatrone, gemcitabine, 5-fluorouracil (5-FU), bleomycin, doxorubicin, daunorubicin, epirubicin, idarubicin, valrubicin, paclitaxel, cabazitaxel, docetaxel, topotecan, irinotecan, etoposide, carboplatin, cisplatin, bortezomib, vinblastine, vincristine, vindesine, vinorelbine, diaziquone, mechlorethamine, mitomycin C, fludarabine, cytosine arabinoside, and combinations thereof.

[0289] Embodiment 125 The use of any of embodiments 97-124, wherein said chemotherapy is immunogenic cell death (ICD)-inducing chemotherapy.

[0290] Embodiment 126 The use of any of embodiments 97-125, wherein said cancer is selected from the group consisting of triple-negative breast cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, classical Hodgkin's lymphoma (cHL), bladder cancer, primary mediastinal B-cell lymphoma (PBMCL), urothelial carcinoma, microsatellite instability-high (MSI-H) solid tumors, mismatch repair-deficient (dMMR) solid tumors, gastric or gastroesophageal junction (GEJ) adenocarcinoma, esophageal squamous cell carcinoma, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, ovarian cancer, anal canal cancer, colorectal cancer, and melanoma.

[0291] Embodiment 127 The use of any of embodiments 97-126, wherein said patient is not administered an immune checkpoint inhibitor concurrently with administration of said CDK4 / 6 inhibitor.

[0292] Embodiment 128 1. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the therapy of triple-negative breast cancer in a patient or patient population selected for extending progression-free survival or overall survival, comprising: The patient or the selection of the patient comprises the steps of: (i) determining whether the triple-negative breast cancer has a surrounding microenvironment that is favorable for immune modulation; (ii) determining whether the chemotherapy regimen is capable of inducing an immune-mediated response; (iii) if both (i) and (ii) are positive, administering an effective amount of a CDK4 / 6 inhibitor prior to administration of said chemotherapy, or optionally prior to said chemotherapy and concurrently with said chemotherapy. comprising The use, wherein the improvement in overall survival or progression-free survival is compared to the overall survival or progression-free survival based on administration of the chemotherapy alone.

[0293] Embodiment 129 The use of embodiment 128, wherein determining whether the cancer has a surrounding microenvironment favorable to immunomodulation comprises comparing the cancer tissue sample with the cancer tissue sample characterized in Figure 7.

[0294] Embodiment 130 The use of embodiment 128, wherein determining whether said cancer has a surrounding microenvironment favorable to immunomodulation comprises evaluating said cancer according to Figure 6.

[0295] Embodiment 131 The use of embodiment 128, wherein determining whether the cancer has a surrounding microenvironment favorable to immunomodulation is according to the Galon immunoscore system.

[0296] Embodiment 132 The use of embodiment 128, wherein determining whether the cancer has a surrounding microenvironment that is favorable for immunomodulation comprises assessing whether the cancer's microenvironment has sufficiently high levels of major histocompatibility complex class I antigens available to elicit an effective immune response.

[0297] Embodiment 133 The use of embodiment 128, wherein determining whether the cancer has a surrounding microenvironment that is favorable for immunomodulation comprises assessing whether the cancer's microenvironment has sufficiently high levels of major histocompatibility complex class II antigens available to elicit an effective immune response.

[0298] Embodiment 134 The use of embodiment 128, wherein determining whether the microenvironment of the cancer has a surrounding microenvironment favorable to immunomodulation comprises assessing whether the cancer has sufficiently high levels of major histocompatibility complex class I and class II antigens available to elicit an effective immune response.

[0299] Embodiment 135 The use of any of embodiments 128, wherein said patient has a cancer that is immunogenically classified as an immunologically hot tumor.

[0300] Embodiment 136 The use of any of embodiments 128, wherein said patient has a cancer that is immunogenically classified as an immunosuppressively altered tumor.

[0301] Embodiment 137 The use of embodiment 128, wherein the patient has a cancer that is a C2 IFN-γ dominant class of cancer, has a cancer with a high IFN-γ signature or a high extended immune signature, or has a cancer that is PD-L1 positive, or a combination thereof.

[0302] Embodiment 138 The use of embodiment 128, wherein determining whether the cancer has a surrounding microenvironment favorable to immunomodulation comprises assessing whether the cancer's microenvironment has a sufficiently high degree of T cell and cytotoxic T cell infiltration.

[0303] Embodiment 139 The use of embodiment 128, wherein determining whether the cancer has a surrounding microenvironment favorable to immune modulation comprises assessing whether the cancer's microenvironment has immune checkpoint activation selected from programmed cell death protein 1 (PD-1) expression and cytotoxic T-lymphocyte-associated antigen 4 (CTLA4) expression.

[0304] Embodiment 140 The use of embodiment 128, wherein determining whether the cancer has a surrounding microenvironment that is favorable for immunomodulation comprises assessing whether the cancer's microenvironment has T-cell immunoglobulin mucin receptor 3 (TIM3) expression and lymphocyte activation gene 3 (LAG3) expression.

[0305] Embodiment 141 The use of embodiment 128, wherein determining whether the cancer has a surrounding microenvironment favorable to immunomodulation comprises assessing whether the microenvironment of the cancer has impaired T cell function.

[0306] Embodiment 142 The use of embodiment 128, wherein determining whether the cancer has a surrounding microenvironment favorable to immune modulation comprises assessing whether the cancer has genomic instability and whether the microenvironment indicates the presence of a pre-existing anti-tumor immune response.

[0307] Embodiment 143 The use of any of embodiments 128-142, wherein said CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof.

[0308] Embodiment 144 The use of any of embodiments 128-142, wherein said CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0309] Embodiment 145 The use of any of embodiments 128-142, wherein said CDK4 / 6 inhibitor is administered within about 24 hours prior to administration of said chemotherapy.

[0310] Embodiment 146 The use of any of embodiments 128-142, wherein said CDK4 / 6 inhibitor is administered within about 4 hours before administration of said chemotherapy.

[0311] Embodiment 147 The use of any of embodiments 128-143, wherein said CDK4 / 6 inhibitor is administered within about 30 minutes prior to administration of said chemotherapy.

[0312] Embodiment 148 The use of any of embodiments 36-147, wherein said chemotherapy is gemcitabine and carboplatin.

[0313] Embodiment 149 1. Use of a compound selected from Compound I, Compound II, Compound III, Compound IV, and Compound V, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for cancer therapy comprising administration of a CDK4 / 6 inhibitor and chemotherapy for a selected patient or patient population in which bone marrow suppression is reduced in human patients undergoing chemotherapy, comprising: The patient or the selection of the patient comprises the steps of: (i) determining whether the cancer has a surrounding microenvironment that is unresponsive to immunomodulation; (ii) determining whether the chemotherapy regimen induces chemotherapy-induced myelosuppression; (iii) if both (i) and (ii) are positive, administering an effective amount of said CDK4 / 6 inhibitor before administration of said chemotherapy, or optionally before and simultaneously with said chemotherapy. comprising The above-mentioned use, wherein the reduction in bone marrow suppression is compared with the bone marrow suppression caused by the administration of the chemotherapy alone.

[0314] Embodiment 150 The use of embodiment 149, wherein determining whether the cancer has a surrounding microenvironment that is not favorable for immune modulation comprises comparing the cancer tissue sample with the cancer tissue sample characterized in Figure 7.

[0315] Embodiment 151 The use of embodiment 149, wherein determining whether said cancer has a surrounding microenvironment that is not favorable for immunomodulation comprises evaluating said cancer according to Figure 6.

[0316] Embodiment 152 The use of embodiment 149, wherein determining whether the cancer has a surrounding microenvironment favorable to immunomodulation comprises assessing the cancer according to the Galon Immunoscore system.

[0317] Embodiment 153 The use of embodiment 149, wherein determining whether the cancer has a surrounding microenvironment that is not favorable for immunomodulation comprises assessing whether the microenvironment of the cancer has low levels of major histocompatibility complex class I antigens.

[0318] Embodiment 154 The use of embodiment 149, wherein determining whether the cancer has a surrounding microenvironment that is not favorable for immunomodulation comprises assessing whether the microenvironment of the cancer has low levels of major histocompatibility complex class II antigens.

[0319] Embodiment 155 The use of embodiment 149, wherein determining whether the cancer has a surrounding microenvironment that is not favorable for immunomodulation comprises assessing whether the cancer's microenvironment has low levels of major histocompatibility complex class I and class II antigens.

[0320] Embodiment 156 The use of embodiment 149, wherein said patient has a cancer that is immunogenically classified as an immunologically cold tumor.

[0321] Embodiment 157 The use of embodiment 149, wherein the patient has a cancer with low IFN-γ expression in the tumor microenvironment, is not of the IFN-γ dominant class of cancer, has a cancer with a low IFN-γ signature or a low extended immune signature, or is PD-L1 negative.

[0322] Embodiment 158 The use of embodiment 149, wherein determining whether the cancer has a surrounding microenvironment that is not favorable for immunomodulation comprises assessing whether the cancer's microenvironment has a low T cell and cytotoxic T cell infiltration.

[0323] Embodiment 159 The use of embodiment 149, wherein determining whether the cancer has a surrounding microenvironment that is not favorable for immunomodulation comprises assessing whether the cancer's microenvironment has low programmed cell death protein 1 (PD-1) expression and low cytotoxic T-lymphocyte-associated antigen 4 (CTLA4) expression.

[0324] Embodiment 160 The use of embodiment 149, wherein determining whether the cancer has a surrounding microenvironment that is not favorable for immune modulation comprises assessing whether the cancer's microenvironment has low expression of T-cell immunoglobulin mucin receptor 3 (TIM3) and low expression of lymphocyte activation gene 3 (LAG3).

[0325] Embodiment 161 The use of any of embodiments 149-160, wherein said CDK4 / 6 inhibitor administered is Compound I, or a pharmaceutically acceptable salt thereof.

[0326] Embodiment 162 The use of any of embodiments 149-160, wherein said CDK4 / 6 inhibitor administered is Compound III, or a pharmaceutically acceptable salt thereof.

[0327] Embodiment 163 The use of any of embodiments 149-160, wherein said CDK4 / 6 inhibitor is administered within about 24 hours prior to administration of said chemotherapy.

[0328] Embodiment 164 The use of any of embodiments 149-160, wherein said CDK4 / 6 inhibitor is administered within about 4 hours before administration of said chemotherapy.

[0329] Embodiment 165 The use of any of embodiments 149-160, wherein said CDK4 / 6 inhibitor is administered within about 30 minutes prior to administration of said chemotherapy.

[0330] Embodiment 166 The use of any of embodiments 149-165, wherein said chemotherapy is selected from cyclophosphamide, trabectedin, temozolomide, melphalan, dacarbazine, oxaliplatin, methotrexate, mitroxatrone, gemcitabine, 5-fluorouracil (5-FU), bleomycin, doxorubicin, daunorubicin, epirubicin, idarubicin, valrubicin, paclitaxel, cabazitaxel, docetaxel, topotecan, irinotecan, etoposide, carboplatin, cisplatin, bortezomib, vinblastine, vincristine, vindesine, vinorelbine, diaziquone, mechlorethamine, mitomycin C, fludarabine, cytosine arabinoside, and combinations thereof.

[0331] Embodiment 167 The use of any of embodiments 149-166, wherein said cancer is selected from the group consisting of triple-negative breast cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, classical Hodgkin's lymphoma (cHL), bladder cancer, primary mediastinal B-cell lymphoma (PBMCL), urothelial carcinoma, microsatellite instability-high (MSI-H) solid tumors, mismatch repair-deficient (dMMR) solid tumors, gastric or gastroesophageal junction (GEJ) adenocarcinoma, esophageal squamous cell carcinoma, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, ovarian cancer, anal canal cancer, colorectal cancer, and melanoma.

[0332] Embodiment 168 The use of any of embodiments 149-166, wherein the cancer is non-small cell lung cancer.

[0333] Embodiment 169 The use of any of embodiments 36-147, wherein said CDK4 / 6 inhibitor is administered one or more times after completion of said chemotherapy treatment in a maintenance treatment regimen, and said chemotherapy is not administered at the time of administration of said CDK4 / 6 inhibitor.

[0334] Embodiment 170 The use of embodiment 169, wherein said CDK4 / 6 inhibitor is administered at a dosing schedule selected from the group consisting of at least once a week, at least once every two weeks, at least once every three weeks, at least once a month, and at least once every six months.

[0335] Embodiment 171 The use of any of embodiments 36-147, wherein said CDK4 / 6 inhibitor is administered one or more times in combination with a chemotherapy agent after completion of standard treatment in a chemotherapy dose-reduced maintenance treatment regimen, wherein said chemotherapy is administered at a dose lower than that administered during standard treatment.

[0336] Embodiment 172 The use of embodiment 171, wherein said CDK4 / 6 inhibitor and said chemotherapy are administered at a dosing schedule selected from the group consisting of at least once per week, at least once every two weeks, at least once every three weeks, at least once per month, at least once every six weeks, at least once every two months, at least once every three months, at least once every four months, at least once every five months, or at least once every six months. [Example]

[0337] Example 1: Trilaciclib improves overall survival and progression-free survival in human metastatic triple-negative breast cancer patients receiving gemcitabine and carboplatin

[0338] Study design Trilaciclib (IV, 240 mg / m) in combination with gemcitabine (IV, 1000 mg / m) plus carboplatin (IV, AUC-2) (G / C) therapy for patients with metastatic TNBC 2 A multicenter, randomized, open-label, phase 2 study (G1T28-04) was designed to investigate the safety, tolerability, efficacy, and PK of once-daily dosing. Patients were randomized (1:1:1) to one of three groups. Group 1: G / C therapy (days 1 and 8 of a 21-day cycle); Arm 2: G / C regimen (days 1 and 8) plus trilaciclib administered IV on days 1 and 8 of a 21-day cycle; Arm 3: G / C regimen (days 2 and 9) plus trilaciclib administered IV on days 1, 2, 8, and 9 of a 21-day cycle Trilaciclib was administered intravenously before GC infusion.

[0339] An overview of the study is shown in Figure 1.

[0340] Tumors were evaluated by immunohistochemistry Price More estrogen and progesterone receptor negative (defined as <10% nuclear staining)Adult patients (≥18 years of age) with evaluable, biopsy-confirmed, locally recurrent or metastatic TNBC (mTNBC) who were HER2-negative according to the American Society of Clinical Oncology / College of American Pathologists practice guidelines (i.e., no overexpression by local immunohistochemistry [0 or 1+] or fluorescence in situ hybridization [HER2 / CEP17 ratio <2·0], or mean HER2 gene copy number <4 signals / nucleus by local evaluation) were eligible for enrollment. Availability of a diagnostic tumor tissue sample confirming TNBC was a prerequisite for enrollment. Hemoglobin levels were ≥9.0 g / dL without red blood cell (RBC) transfusions within 14 days prior to the first dose of trilaciclib, and absolute neutrophil count (ANC) ≥1.5 × 10 9 / L and total platelet count ≥ 100 × 10 9 / L. Patients were ineligible if they had received more than two prior cytotoxic chemotherapy regimens for locally recurrent TNBC or mTNBC. Chemotherapy administered in the neoadjuvant / adjuvant setting was considered a single line of therapy if there was less than 12 months between the last treatment and disease recurrence. Patients had to have Eastern Cooperative Oncology Group performance status of 0 or 1 and adequate renal and hepatic function as determined by laboratory tests of serum creatinine (≤1.5 mg / dL or creatinine clearance ≥60 mL / min), total bilirubin ≤1.5 × upper limit of normal (ULN), and aspartate transaminase and alanine transaminase ≤2.5 × ULN (or ≤5 × ULN if liver metastases were present). Except for alopecia, no significant changes from previous treatment were noted. Non Hematologic toxicity grade ≤1 recovery Patients were required to have had a malignancy other than TNBC within 3 years prior to randomization, central nervous system metastasis / leptomeningeal disease requiring immediate treatment, or uncontrolled Ischemic Patients were ineligible for inclusion if they had cardiac disease or symptomatic congestive heart failure, a known history of stroke or cerebrovascular accident within 6 months before the first dose of trilaciclib, a known severe active infection, or any other uncontrolled severe chronic illness or psychiatric condition that would affect patient safety, compliance, or follow-up. Before participating in the test,Prior radiation therapy or cytotoxic chemotherapy required a washout period of 2 or 3 weeks, respectively.

[0341] The study was designed and conducted in accordance with the principles of the Declaration of Helsinki and the International Conference on Harmonization's Good Clinical Practice guidelines. The study protocol and all study-related materials were approved by the Institutional Review Board or independent ethics committee at each study site. Written informed consent was obtained from each patient before the start of study procedures.

[0342] Patients were randomly assigned to the following given treatments in 21-day cycles: Group 1 received gemcitabine and carboplatin on days 1 and 8 (chemotherapy alone); Group 2 received trilaciclib before gemcitabine and carboplatin on days 1 and 8 (trilaciclib + chemotherapy on days 1 and 8); and Group 3 received trilaciclib only on days 1 and 8 and trilaciclib before gemcitabine and carboplatin on days 2 and 9 (trilaciclib on days 1 and 8, trilaciclib + chemotherapy on days 2 and 9). Group 3 was included to test the hypothesis that a second dose of trilaciclib before chemotherapy could increase the proportion of blood stem and progenitor cells that are temporarily in a suspended state during chemotherapy administration, thereby improving myelosuppressive outcomes. Gemcitabine was administered at 1000 mg / m 2 Both were administered intravenously at a carboplatin concentration-time area under the curve (AUC) of 2 μg × h / mL. 2 Trilaciclib was administered intravenously 30 minutes (tolerance range 25-35 minutes) before gemcitabine and carboplatin treatment. No dose modifications of trilaciclib were permitted.

[0343] Treatment cycles were performed consecutively without interruption unless necessary for toxicity control. If dose reductions were required for chemotherapy, they were performed in the following order: first, gemcitabine dose was reduced to 1000 mg / m 2 to 800 mg / m 2Second, the carboplatin dose was reduced from AUC 2 μg × h / mL to AUC 1.5 μg × h / mL; third, either carboplatin or gemcitabine was discontinued while the other drug was continued at a reduced dose; and finally, all study drugs were permanently discontinued. Dose reductions were allowed only once per cycle and were permanent.

[0344] Trilaciclib was administered only in combination with GC therapy, and was maintained or discontinued if chemotherapy was maintained or discontinued. Study drug treatment continued until disease progression, unacceptable toxicity, consent withdrawal, or investigator discontinuation, whichever occurred first.

[0345] Per protocol, samples for hematological laboratory tests were collected on days 1, 8, and 15 of each 21-day cycle, regardless of treatment group. If the start of the next cycle was delayed, laboratory tests were performed weekly (e.g., on days 22, 29, 36, etc.) until the patient was able to begin the next cycle or chemotherapy was permanently discontinued. Unscheduled laboratory tests were permitted if clinically indicated. The use of prophylactic growth factors, including granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF), was not permitted during cycle 1. Otherwise, supportive care, including transfusions, was permitted throughout treatment as needed. Platelets were transfused at thresholds of 10,000 / μL or less or a total platelet count of 50,000 / μL (100,000 / μL in the case of central nervous system or ocular hemorrhage). Patients with hemoglobin levels below 8.0 g / dL or symptomatic anemia were treated with red blood cell transfusions at the investigator's discretion. The percentage of patients receiving red blood cell transfusions and the number of red blood cell transfusions received over time were analyzed from day 1 of the study as part of a sensitivity analysis beyond week 5.

[0346] Antitumor response was assessed by the investigator according to Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1. For tumor assessment, computed tomography or magnetic resonance imaging was performed at screening and at protocol-specified intervals (every 9 weeks for the first 6 months, then every 12 weeks) until disease progression, withdrawal of consent, or receipt of subsequent anticancer treatment. Bone and brain scans were required at screening. Follow-up evaluation of bone lesions could use a different imaging method; brain scans were repeated as part of tumor assessment only if brain metastases were present.

[0347] To assess the cytokine-producing capacity of T cells, whole blood was stimulated overnight (15–18 h) with 5 μg / mL staphylococcal endotoxin B in the presence of brefeldin A. Cells were processed and labeled with fluorophore-conjugated antibodies against IFN-γ, IL-17A, CD3, and CD8, and evaluated by flow cytometry (BD FACSCalibur and FACSCanto II clinical cell analyzers; BD Biosciences, Franklin Lakes, NJ, USA) by Covance Central Laboratory Services (Indianapolis, IN, USA and Geneva, Switzerland). Flow cytometry data were analyzed by Fios Genomics.

[0348] Two established signatures (Prosigna Breast Cancer Prognostic Gene Signature Assay [PAM50] and Lehmann type 1-4 triple-negative breast cancer) were used to classify patient tumors as CDK4 / 6-independent, dependent, or indeterminate. Because triple-negative breast cancer is primarily a functionally CDK4 / 6-dependent disease, despite a genomic retinoblastoma inactivation rate of only 20%, these signatures were selected to provide a more comprehensive analysis of CDK4 / 6 sensitivity. Using the PAM50 signature, CDK4 / 6 independence correlates with basal-like tumors. Because of unclear or heterogeneous dependence on the CDK4 / 6 pathway for growth, the remaining PAM50 signature groups (including HER2, normal-like, luminal A, and luminal B) are classified as CDK4 / 6-indeterminate. Conversely, using the Lehmann signature, CDK4 / 6 dependence correlates closely with luminal androgen receptor tumors, while the remaining Lehmann signature group (including basal-like and mesenchymal) is classified as CDK4 / 6 indeterminate for the same reasons outlined for the PAM50 signature.

[0349] Safety was monitored continuously from the time of informed consent until 30 days after the last dose of study treatment. Safety assessments included analysis of treatment duration and dose modifications, evaluation of treatment-emergent adverse events and treatment-emergent serious adverse events, infusion reactions, clinical safety assessments, vital signs, physical examinations, and electrocardiograms. Treatment-emergent adverse events were summarized according to grade (Common Terminology Criteria for Adverse Events [CTCAE] version 4.03) and relationship to study drug. A serious adverse event was defined as any untoward medical occurrence, regardless of dose, that resulted in death, a life-threatening event (i.e., the patient was at risk of death at the time of the event), hospitalization or prolongation of current hospitalization, persistent or significant disability or impairment, or congenital anomaly or birth defect.

[0350] Because the primary toxicity of chemotherapy is myelosuppression (which trilaciclib is hypothesized to mitigate), several hematologic parameters were assessed across multiple hematopoietic lineages, including the incidence and severity of hematologic adverse events, laboratory values ​​(absolute neutrophil count, hemoglobin concentration, and platelet count), supportive care interventions (red blood cell and platelet transfusions, G-CSF use), and dose intensity and incidence of gemcitabine and carboplatin dose reductions. A complete description of all parameters is further described below.

[0351] Outcome The primary objective was to evaluate the safety and tolerability of trilaciclib administered with chemotherapy; endpoints of particular interest were detailed in the statistical analysis plan, which defined the primary endpoint as severe neutropenia in cycle 1 (severe neutropenia defined as Common Terminology Criteria for Adverse Events grade 4, absolute neutrophil count <0.5 × 10 9 Duration of severe neutropenia in Cycle 1 was defined as an initial absolute neutrophil count of 0.5 × 10 cells / L or greater and the presence of severe neutropenia during treatment. 9 From days of < 10 cells / L, initial absolute neutrophil count 0.5 × 10 9 cells / L or more until the end of the cycle (0.5 × 10 9 Duration of severe neutropenia was defined as the number of days without an absolute neutrophil count of less than 0.5 × 10 cells / L during treatment. Patients who did not experience severe neutropenia in cycle 1 were assigned a duration of severe neutropenia of 0.5 × 10 cells / L. 9 The endpoint was binary, defined as one or more measurements of < 0.5 × 10 cells / L. Both scheduled and unscheduled blood test results were included in the analyses for both primary endpoints. For the primary analysis, which was based on the clinical association between severe neutropenia and increased risk of infection and morbidity and mortality, a clinically meaningful 0.5 × 10 9 The level of cells / L was selected.

[0352] Key secondary endpoints included the occurrence of red blood cell transfusions after week 5, G-CSF administration, platelet transfusions, and overall survival. Red blood cell transfusions before week 5 were excluded based on the half-life of red blood cells (approximately 8 to 9 weeks) to ensure that analyses of potential benefit were not confounded by the effects of residual prior treatment. The occurrence of red blood cell and platelet transfusions was a binary endpoint (yes or no), and total transfusions was a count endpoint (transfusions with a unique start date). Overall survival was calculated as the time (in months) from the date of randomization to the date of death, regardless of course.

[0353] Supporting secondary antitumor activity endpoints were the proportion of patients achieving an objective response (defined as a confirmed complete or partial response), duration of response, and progression-free survival. Clinical benefit rates were calculated using data from patients who had a complete or partial response at any time posttreatment or stable disease for at least 24 weeks. Patients without a complete or partial response and an uncertain duration of stable disease were considered unevaluable. Progression-free survival was defined as the time (in months) from the date of randomization to radiologically confirmed disease progression or death from any cause, whichever occurred first.

[0354] statistical analysis Consistent with the trilaciclib development program, specific endpoints prespecified in the statistical analysis plan were used to demonstrate superiority of Arm 3 over Arm 1 for at least one primary endpoint (either duration of severe neutropenia or occurrence of severe neutropenia in Cycle 1) with 90% power. Using the equally weighted Bonferroni method to maintain an overall two-sided type I error rate of 0.05, 64 patients (32 per arm) were required to detect a 3-day reduction in the duration of severe neutropenia in Cycle 1, calculated as a common SD of 2.5 days or a 41-percentage-point absolute reduction in the proportion of patients with severe neutropenia (i.e., 45% in Arm 1 and 4% in Arm 3). Assuming a 5% discontinuation rate, a total of 102 patients (34 per arm) were required.

[0355] Nonparametric analysis of covariance was used to assess treatment group differences in the duration of severe neutropenia in cycle 1, with baseline absolute neutrophil count as a covariate and stratification factors and treatment as fixed effects. Treatment effects were assessed for the occurrence of severe neutropenia, G-CSF administration, red blood cell transfusions after week 5, and platelet transfusions using a modified Poisson regression model. This model included the same fixed items used for the duration of severe neutropenia and used baseline absolute neutrophil count as a covariate for the severe neutropenia and G-CSF administration analyses, baseline hemoglobin concentration as a covariate for the red blood cell transfusion analyses, and baseline platelet count as a covariate for the platelet transfusion analyses. The model adjusted for treatment duration (weeks). The percentage of patients receiving red blood cell transfusions and the number of red blood cell transfusions over time from study day 1 were analyzed as part of sensitivity analyses. Treatment effects were assessed for the number of red blood cell transfusions and platelet transfusions after week 5 using a negative binomial regression model. The model included the same fixed items used for the duration of severe neutropenia, with baseline hemoglobin as a covariate for the red blood cell transfusion analysis and baseline platelets as a covariate for the platelet transfusion analysis. The model adjusted for treatment duration (weeks). The number of dose reductions for any reason was analyzed using a negative binomial regression model that included only stratification factors and treatment as fixed effects and adjusted for the number of cycles. A Hochberg-based gatekeeping method was used to control for a family-wise error rate of 0.025 (one-sided) for the primary endpoint of myelosuppression and key secondary endpoints. Model-based point estimates are reported with 95% CIs.

[0356] Treatment group differences in objective response were analyzed using the exact Cochran-Mantel-Haenszel test, which accounts for stratification factors. The 95% CI for the proportion of patients achieving an objective response was calculated using the exact Clopper-Pearson method. For time-to-event variables, such as duration of response, time to progression, and overall survival, the median duration and its 95% CI were estimated using the Kaplan-Meier method. Treatment group differences were tested using the stratified log-rank test to account for stratification factors. Hazard ratios (HRs) and their associated 95% CIs were calculated from Cox proportional hazards models, with treatment and stratification factors as fixed effects.

[0357] The statistical analysis plan prespecified the primary statistical comparison for the primary and key secondary endpoints as between arm 3 and arm 1, and the secondary comparisons as between arm 2 and arm 1 and between the combined trilaciclib arms and arm 1.

[0358] Activity analyses were performed using the intention-to-treat (ITT) population based on assigned treatment for myelosuppression and antitumor activity endpoints, except for tumor response endpoints (objective response and clinical benefit), which were analyzed in patients who received at least one dose of study drug, had measurable target lesions at baseline tumor assessment, and either underwent at least one post-treatment tumor assessment (or had no post-treatment tumor assessment but clinical progression documented by the investigator), or died of disease progression before the first post-treatment tumor scan (response-evaluable population). Survival follow-up was calculated from the date of randomization to the date of death or last contact at the designated activity data cutoff throughout. Safety analyses included all patients who received at least one dose of study drug. Stratification factors (number of prior systemic therapies and hepatic involvement) were adjusted for in the statistical models.

[0359] Prespecified subgroup analyses were performed for progression-free survival and overall survival to assess the consistency of treatment effects (i.e., age group, race, liver involvement, country, ECOG performance status, number of prior therapies, BRCA classification, and histological triple-negative breast cancer classification).

[0360] To assess the theoretical risk that trilaciclib may reduce antitumor activity in patients with CDK4 / 6-dependent tumors by shutting down CDK4 / 6-dependent tumor cells during chemotherapy, we performed other prespecified subgroup analyses of antitumor activity endpoints (objective response, progression-free survival, and overall survival) using two established signatures (PAM50 and Lehmann triple-negative breast cancer types 1–4) to classify patient tumors as CDK4 / 6-independent, dependent, or indeterminate.

[0361] Post-hoc analyses of antitumor activity endpoints (objective response, progression-free survival, and overall survival) were performed based on the overall median number of cycles patients received during the study. Analyses were performed for patients categorized by whether they received 1 to 7 cycles or ≥ 7 cycles.

[0362] result One hundred forty-two patients were screened, and 102 eligible patients were randomly assigned to chemotherapy alone (Group 1; n = 34), trilaciclib plus chemotherapy (Group 2; n = 33), or trilaciclib (days 1 and 8) or trilaciclib plus chemotherapy (days 2 and 9) (Group 3; n = 35; intention-to-treat population). Of these, 98 (96%) patients received at least one dose of study drug (safety analysis population). Baseline demographic characteristics were comparable between treatment groups and are disclosed in Table 3. Of the 102 patients, 38 (37%) had received one or two prior chemotherapy treatments, and 26 (25%) had liver metastases.

[0363] [Table 3]

[0364] The addition of trilaciclib to gemcitabine and carboplatin did not result in a significant improvement in the prespecified primary myelosuppression endpoint. During cycle 1, the mean duration of severe neutropenia was 1 day (SD 2.4) in group 1, 2 days (3.5) in group 2, and 1.0 day (2.6) in group 3 (p=0.70). Severe neutropenia occurred in 9 of 34 patients (26%) in group 1, 12 of 33 patients (36%) in group 2, and 8 of 35 patients (23%) in group 3 (p=0.70; Table 2). Over 100 weeks, the number of red blood cell transfusions after week 5 was reduced in both trilaciclib groups (4.6 in group 1 vs. 1.9 in group 2 and 1.6 in group 3; p=0.020). Red blood cell transfusion data collected on study day 1 (sensitivity analysis) were similar to those observed when data prior to week 5 were excluded. No significant differences were observed in the number of patients receiving G-CSF or platelet transfusions.

[0365] [Table 4]

[0366] At the last assessment of drug exposure (data cut June 28, 2019), the number of patients who had at least one carboplatin dose reduction was 10 (33%) in group 1, 13 (39%) in group 2, and 15 (43%) in group 3. For gemcitabine, 13 (43%) in group 1, 20 (61%) in group 2, and 17 (49%) in group 3 had at least one dose reduction. The addition of trilaciclib to gemcitabine and carboplatin increased the duration and cumulative dose of gemcitabine and carboplatin exposure compared with patients treated with gemcitabine and carboplatin alone. The median treatment duration was 101 days (IQR 63-203 [median after 4 cycles]) in group 1, 161 days (77-287 [median after 7 cycles]) in group 2, and 168 days (91-217 [median after 8 cycles]) in group 3. The median cumulative carboplatin dose was AUC 15 μg × h / mL (IQR 8-28) in group 1, AUC 24 μg × h / mL (IQR 10-40) in group 2, and AUC 22 μg × h / mL (IQR 15-34) in group 3. For gemcitabine, the median cumulative dose was 7306.2 mg / m in group 1. 2 (IQR 4020.1-15138.9) and 12000.0 mg / m in Group 2. 2 (IQR 5029.4-21882.7), and 11800.1 mg / m in group 3. 2 Despite the longer duration of gemcitabine and carboplatin in patients receiving trilaciclib, the incidence of treatment-emergent hematologic adverse events in the trilaciclib group was equal to or less than that in the chemotherapy-only group.

[0367] At the time of the last safety data assessment (data cut May 15, 2020), all patients except one (group 3) had one or more treatment-emergent adverse events. In most patients, treatment-emergent adverse events were considered drug-related. The most common treatment-emergent adverse events were anemia (22 of 34 [73%]), neutropenia (21 [70%]), and thrombocytopenia (18 [60%]) in group 1; neutropenia (27 of 33 [82%]), thrombocytopenia (19 [58%]), and anemia (17 [52%]) in group 2; and neutropenia (23 of 35 [66%]), thrombocytopenia (22 [63%]), and nausea (17 [49%]) in group 3. Febrile neutropenia occurred in one patient in group 1 and one patient in group 2. Treatment-emergent serious adverse events were reported in 10 (33%) patients in Group 1, 11 (33%) in Group 2, and 4 (11%) in Group 3. Fewer than two patients experienced treatment-emergent serious adverse events. A total of 58 deaths occurred: 25 in Group 1 (disease progression [n = 21], treatment-emergent adverse event [n = 1; right ventricular failure unrelated to treatment], other [n = 3]), 13 in Group 2 (disease progression [n = 11], other [n = 2]), and 20 in Group 3 (disease progression [n = 19], other [n = 1]). Overall, the number of patients reporting treatment-emergent adverse events leading to discontinuation of study drug was similar in each group: 10 (33%) in Group 1, 14 (42%) in Group 2, and 11 (31%) in Group 3.

[0368] At the time of the last assessment of antitumor efficacy (data cut May 15, 2020), the median follow-up was 8.4 months (IQR 3.8-15.6) in Arm 1, 14 months (5.5-26.8) in Arm 2, and 15.3 months (6.7-23.7) in Arm 3. Among patients evaluable for response, the proportion of patients who achieved an objective response was 29.2% (7 of 24) in Arm 1, 50% (15 of 30) in Arm 2, and 15.3% (6 of 23) in Arm 3. 38.7%The proportion of patients who achieved clinical benefit, including stable disease for ≥24 weeks, was 38% (9 of 24) in group 1, 57% (17 of 30) in group 2, and 43% (13 of 30) in group 3. The median progression-free survival was 9.4 months (IQR 5.3-13.0) in group 2, 7.3 months (IQR 6.2-13.9) in group 3, and 5.7 months (IQR 2.2-9.9) in group 1. The HRs for both groups, analyzed relative to group 1, were 0.62 (95% CI 0.32-1.20; p=0.21) in group 2 and 0.63 (95% CI 0.42-1.20; p=0.21) in group 3. CI Patients enrolled in both trilaciclib groups showed a significant improvement in overall survival compared with patients enrolled in group 1 (median not reached [IQR 9.4 to not reached] and HR 0.31, 95% CI 0.15 to 0.63; p = 0.001 in group 2). 6; In group 3, mean survival was 17.8 months [IQR 8.8-32.7] and HR 0.40, 95% CI 0.22-0.74; p=0.0004] compared with 12.6 months [IQR 5.8-17.8] in group 1 (Table 5; Figure 2). Results of pooled subgroup analyses indicated that, at the data cutoff of June 28, 2019, the observed benefits in progression-free survival (Figure 4B) and overall survival (Figure 4A) were consistent across subgroups. Updated analyses between groups 1 and 3 are shown in Figure 4C and Figure 4D.

[0369] Prespecified evaluation of objective response, progression-free survival, and overall survival between and within groups 1, 2, and 3 for tumors classified as CDK4 / 6-independent, -dependent, or indeterminate revealed no consistent trend in favor of one tumor subtype over another.

[0370] [Table 5]

[0371] The addition of trilaciclib to gemcitabine and carboplatin did not protect lymphocyte counts or enhance T-cell activation, but in a prespecified analysis, patients treated with gemcitabine and carboplatin plus trilaciclib had a higher T-cell count than patients in arm 1. ex vivo A high frequency of CD8+ T cells producing IFN-γ was observed after stimulation (Fig. 5).

[0372] Archival tumor tissue from TNBC diagnoses was analyzed for two distinct published RNA signatures: 1) CDK4 / 6-independent and variable CDK4 / 6-dependent buckets defined by PAM50 (see Prat et al., Response and survival of breast cancer intrinsic subtypes following multi-agent neoadjuvant chemotherapy. BMC Med. 2015;13:303. doi: 10.1186 / s12916-015-0540-z, incorporated herein by reference); and 2) Lehmann (see Lehmann et al., Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies. J Clin Invest. 2011;121:2750-67. doi: 10.1186 / s12916-015-0540-z, incorporated herein by reference). The CDK4 / 6-dependent and variable CDK4 / 6-dependent buckets defined by the NIH standard deviation (see NIH standard deviation).

[0373] [Table 6]

[0374] The selection of trilaciclib in chemotherapy regimens did not antagonize chemotherapy efficacy in TNBC patients with variable or CDK4 / 6-dependent tumors, i.e., tumors classified as PAM50 Other (Her2, normal-like, LumA, LumB) or Lehmann TNBC type-4 LAR (Table 7). There was no difference in ORR / PFS / OS in known independent (basal-like) patients between treatments.

[0375] [Table 7]

[0376] The inclusion of trilaciclib in chemotherapy regimens did not antagonize chemotherapy efficacy in TNBC patients with variable and CDK4 / 6-independent tumors, i.e., tumors classified as PAM50 Basal-Like or Lehmann TNBC type-4 Other (BL1, BL2, M) (Table 8).

[0377] [Table 8]

[0378] Results from a post-hoc analysis of antitumor efficacy according to median number of cycles (1-7 vs. >7 cycles) are shown in Table 9. In all three treatment groups, the proportion of patients who achieved an objective response was higher in patients who received more than 7 cycles compared with patients who received 7 or fewer cycles.

[0379] [Table 9]

[0380] Example 2: Ayers Immunoscore Analysis Tumor samples from patients participating in the clinical trial described in Example 1 were analyzed by Q 2The assays were performed by Bioscience Solutions (Morrisville, NC), and their Ayers immune scores were determined according to Ayers et al., IFN-γ-related mRNA Profile Predicts Clinical Response to PD-1 Blockade, J Clin Invest. 2017127(8)2930-2940. Data were processed using RNA Access, and FPKM normalized, followed by log10 transformation, and averaged.

[0381] 89 samples were analyzed. The calculated signature scores for both the IFN-γ signature and the extended immune signature were unimodal, and the median score was used to define the "high" and "low" categories. Figure 8A shows the distribution of the Ayers' IFN-γ signature for the 89 samples tested. Figure 8B shows the distribution of the Ayers' extended immune signature for the 89 samples tested.

[0382] Survival and response rates between treatment groups within predefined immune response groups were determined using a series of pairwise two-group tests based on data derived from the G1T28-04 clinical trial described in Example 1. Additionally, analyses were performed to examine differences in survival and response rates between different immune response categories within a single treatment group. The results are shown in Tables 10 and 11.

[0383] [Table 10]

[0384] [Table 11]

[0385] Kaplan-Meier curves were generated to visualize overall survival and progression-free survival (PFS) between groups (Figures 8C–8F and 9A–9D). The groups were divided as follows: Group 1 (gemcitabine plus carboplatin administered only on days 1 and 8 of a 21-day cycle), Group 2 (gemcitabine plus carboplatin plus trilaciclib administered on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine plus carboplatin administered on days 2 and 9 of a 21-day cycle, trilaciclib administered on days 1, 2, 8, and 9 of a 21-day cycle), and Group 4 (Group 2 plus Group 3) for the Kaplan-Meier curves.

[0386] As shown, patients with TNBC who received trilaciclib as part of their dosing regimen and had a "high" IFN-γ signature score and / or a "high" expanded immune signature score had significantly improved overall survival compared to patients with TNBC who did not receive trilaciclib as part of their dosing regimen and had a "high" IFN-γ signature score and / or a "high" expanded immune signature score (p=0.0194; p=0.036, respectively).

[0387] Example 3: Six-class immunoclassification analysis of Thorsson et al. Tumor samples from patients participating in the clinical trial described in Example 1 (Clinical trials.gov ID NCT02978716) were analyzed by Q 2 The cells were assayed by BioLegend Solutions (Morrisville, NC) and their 6-class immunoclassification determined as described in Thorsson V, et al. "The Immune Landscape of Cancer." Immunity, vol. 51, no. 2, 2018, pp. 812-830. doi: 10.1016 / j.immuni.2018.03.023 (the entire contents of which are incorporated herein by reference).

[0388] Briefly, a six-step method was implemented to apply the Thorsson et al. classification to 89 pretreatment triple-negative breast cancer samples from the clinical trial described in Example 1. After RNAseq data acquisition, data cleaning was performed and homogenized by reconciling gene and sample names from the data source. Batch correction was performed to equate the data generated by the clinical trial and the TCGA data to ensure valid classification. Briefly, samples in the resulting TCGA expression data were randomly downsampled to more closely represent the abundance of PAM50 classes in the clinical trial data (previously derived, see Example 1, Table 7). To estimate batch effects in the clinical trial data, linear regression modeling of each gene on log2-transformed upper quartile-normalized representations was used. These estimated batch effects were then removed by subtraction from the representations of the clinical trial samples, yielding a mean shift toward the TCGA samples.

[0389] A PCA plot was used to examine the validity of this approach in reconciling the dataset. Before correction, the clinical trial samples and TCGA samples exhibited clear batch effects due to separation. The correction procedure described above eliminated the separation between the two sample groups and made the two sets of expression data comparable. After batch correction, the derived data was input into the Gibbs immune clustering software package (available at https: / / github.com / CRI-iAtlas / ImmuneSubtypeClassifier) ​​to classify the clinical trial samples with respect to Thorsson's six-class immune response scheme. The script and its requirements were downloaded, installed, and run on the batch-corrected clinical trial data.

[0390] The distribution of classes seen in the clinical trial samples is shown in Table 12.

[0391] [Table 12]

[0392] No patients were classified as immunologically silent (C5). This is not surprising, since none of the training samples used by Thorsson et al. for breast cancer were originally classified as immunologically silent. Furthermore, C1 and C2 other than The low prevalence of these classes led to the exclusion of tests for C3 vs. non-C3, C4 vs. non-C4, and C6 vs. non-C6.

[0393] Survival and response rates between treatment groups within predefined immune response groups were determined using a series of pairwise two-group tests based on data derived from the G1T28-04 clinical trial described in Example 1. Additionally, analyses were performed to examine differences in survival and response rates between different immune response categories within a single treatment group. The results are shown in Table 13.

[0394] [Table 13]

[0395] Kaplan-Meier curves were generated to visualize overall survival and progression-free survival between groups (see Figures 10A–10D). The groups were divided as follows: Group 1 (gemcitabine + carboplatin administered only on days 1 and 8 of a 21-day cycle), Group 2 (gemcitabine + carboplatin + trilaciclib administered on days 1 and 8 of a 21-day cycle), Group 3 (gemcitabine + carboplatin administered on days 2 and 9 of a 21-day cycle, trilaciclib administered on days 1, 2, 8, and 9 of a 21-day cycle), and Group 4 (Group 2 + Group 3) for the Kaplan-Meier curves.

[0396] As shown, patients with TNBC classified as C2 IFN-γ dominant who received trilaciclib as part of their dosing regimen had significantly improved overall survival compared to patients with TNBC classified as C2 IFN-γ dominant who did not receive trilaciclib as part of their dosing regimen (p=0.036).

[0397] Example 4: PD-L1 Tumor Status Patient tumors from the G1T28-04 clinical trial described in Example 1 were characterized based on their PD-L1 expression score as negative or positive if <1% or ≥1% of the total tumor area contained PD-L1-labeled immune cells, respectively, using the Ventana SP142 assay. The association between PD-L1 expression and antitumor efficacy was assessed using proportional hazards regression. The groups were divided as follows: Group 1 (gemcitabine + carboplatin administered only on days 1 and 8 of a 21-day cycle), Group 2 (gemcitabine + carboplatin + trilaciclib administered on days 1 and 8 of a 21-day cycle), and Group 3 (gemcitabine + carboplatin administered on days 2 and 9 of a 21-day cycle, and trilaciclib administered on days 1, 2, 8, and 9 of a 21-day cycle). The results are shown in Table 14.

[0398] [Table 14]

[0399] As shown above, patients with PD-L1-positive TNBC tumors who received trilaciclib had a statistically significant overall survival (p=0.005) compared with patients with PD-L1-positive TNBC tumors who did not receive trilaciclib.

[0400] Example 5: Trilaciclib administration enhances T cell proliferation A phase 2 trial of carboplatin, etoposide, and atezolizumab with or without trilaciclib has begun in patients with previously untreated extensive-stage small cell lung cancer (Clinicaltrials.gov ID NCT03041311). The carboplatin trial included a 21-day induction phase and a 21-day maintenance phase. Four induction phase cycles were completed before the start of the maintenance phase cycle.

[0401] During the induction phase, patients received trilaciclib (240 mg / m² diluted in 250 mL D5W or 0.9% sodium chloride solution) or placebo (250 mL D5W or 0.9% sodium chloride solution) administered intravenously once daily on days 1–3 of each 21-day cycle of etoposide / carboplatin / atezolizumab (E / P / A) therapy (maximum of four cycles total). The carboplatin dose was calculated using the Calvert formula [total carboplatin dose (mg) = (AUC target) × (GFR + 25)], where AUC target = 5 (maximum 750 mg) IV over 30 minutes on day 1 of each 21-day cycle, and 100 mg / m² etoposide administered intravenously over 60 minutes once daily on days 1, 2, and 3. Atezolizumab (1200 mg) in 250 mL of 0.9% sodium chloride solution was administered as an IV infusion on day 1 of each 21-day cycle during both the induction and maintenance phases. Atezolizumab was infused over 60 minutes for the first dose, and if tolerated, all subsequent infusions were delivered over 30 minutes. Atezolizumab was administered after completion of Compound I or placebo, etoposide, and carboplatin.

[0402] To identify biomarkers of immune response and immunomodulatory activity between the trilaciclib and placebo arms, we analyzed the TCRB locus in blood samples. All samples were sequenced using the 1-rxn TCRB assay. The number of expanded T cell clones was determined by differential abundance analysis of T cell receptor β sequences in whole blood obtained from patients after induction therapy and before the start of maintenance therapy relative to baseline. Trilaciclib responders had more clonal expansion than placebo responders (p=0.01) and more clonal expansion than trilaciclib non-responders (p=0.006), suggesting that increased clonal expansion is a biomarker for both trilaciclib MOA and clinical response (Figures 11 and 12).

[0403] Furthermore, trilaciclib-treated responders also generated more de novo proliferative clones (p = 0.001, Figure 13) and, significantly, significantly increased the ratio of de novo proliferative clones to total T-cell clones compared with treatment-treated patients who did not receive trilaciclib (p = 0.001, Figure 14). Stratification of patients below and above the median ratio of de novo proliferative clones to total T-cell clones revealed a nonsignificant trend toward longer overall survival (OS) in patients with higher levels of peripheral T-cell clonal expansion (HR [95% CI] = 0.56, P = 0.10). Subgroup analysis (trilaciclib vs. placebo) revealed that patients above the median ratio of de novo proliferative clones had significantly longer OS when treated with trilaciclib (HR = 0.34; p = 0.04), with a similar nonsignificant trend toward longer PFS. Similar benefits were observed for median clonal expansion and median de novo proliferative clone stratification. Unlike placebo, trilaciclib significantly increased the number and proportion of newly proliferating clones, indicating that adding trilaciclib to an etoposide, carboplatin, and atezolizumab regimen enhances T cell-mediated antitumor responses.

[0404] The specification has been described with reference to embodiments of the present invention. However, those skilled in the art will recognize that various modifications and changes can be made without departing from the scope of the invention as set forth in the claims below. The specification is therefore to be regarded in an illustrative rather than a limiting sense, and all such modifications are intended to be included within the scope of the present invention.

Claims

1. 2. A method for cancer therapy for a patient or patient population selected for extended progression free survival or overall survival comprising administering to said patient a compound having the following structure: 【Chemistry 1】 or a pharmaceutically acceptable salt thereof, The method comprises the steps of: (i) determining whether the patient's cancer has a surrounding microenvironment that is favorable for immune modulation; (ii) determining whether the patient is amenable to an immunogenic cell death (ICD)-inducing chemotherapy regimen; (iii) if it is determined that the patient's cancer benefits from immunomodulation and that the patient can be administered the ICD-guided chemotherapy regimen; (a) administering to said patient an effective amount of said CDK4 / 6 inhibitor; (b) administering to said patient an effective amount of said ICD-guided chemotherapy regimen. comprising the CDK4 / 6 inhibitor is administered prior to administration of the ICD-inducing chemotherapy; The pharmaceutical composition, wherein administration of the CDK4 / 6 inhibitor and the ICD-inducing chemotherapy regimen to the patient results in an extension of progression-free survival or overall survival compared to a patient with cancer who has been administered ICD-inducing chemotherapy alone without a CDK4 / 6 inhibitor.

2. The pharmaceutical composition of claim 1, wherein the determination of whether the cancer has a surrounding microenvironment favorable for immunomodulation is according to the Galon immunoscore system.

3. 2. The pharmaceutical composition of claim 1, wherein determining whether the cancer has a surrounding microenvironment favorable to immunomodulation comprises assessing whether the cancer has sufficiently high levels of major histocompatibility complex class I antigens available to elicit an effective immune response.

4. 2. The pharmaceutical composition of claim 1, wherein determining whether the cancer has a surrounding microenvironment favorable to immunomodulation comprises assessing whether the cancer has sufficiently high levels of major histocompatibility complex class II antigens available to elicit an effective immune response.

5. 2. The pharmaceutical composition of claim 1, wherein determining whether the cancer has a surrounding microenvironment favorable to immunomodulation comprises assessing whether the cancer has sufficiently high levels of major histocompatibility complex class I and class II antigens available to elicit an effective immune response.

6. The pharmaceutical composition of any one of claims 1 to 5, wherein the patient has a cancer that is immunogenically classified as an immunologically hot tumor.

7. The pharmaceutical composition of any one of claims 1 to 5, wherein the patient has a cancer that is immunogenically classified as an immunosuppressively altered tumor.

8. 6. The pharmaceutical composition of any one of claims 1 to 5, wherein determining whether the cancer has a surrounding microenvironment favorable to immune modulation comprises assessing whether the cancer is of an IFN-γ dominant class of cancer, whether it has a cancer microenvironment with a high IFN-γ signature, or whether it has a cancer microenvironment with a high extended immune signature, PD-L1 positivity, or a combination thereof.

9. The pharmaceutical composition of any one of claims 1 to 8, wherein the CDK4 / 6 inhibitor is administered within about 24 hours prior to administration of the ICD-inducing chemotherapy.

10. The pharmaceutical composition of any one of claims 1 to 8, wherein the CDK4 / 6 inhibitor is administered within about 4 hours before administration of the ICD-inducing chemotherapy.

11. 9. The pharmaceutical composition of any one of claims 1 to 8, wherein the CDK4 / 6 inhibitor is administered within about 30 minutes before administration of the ICD-inducing chemotherapy.

12. 12. The pharmaceutical composition of any one of claims 1 to 11, wherein the ICD-guided chemotherapy is selected from the group consisting of cyclophosphamide, trabectedin, temozolomide, melphalan, dacarbazine, oxaliplatin, methotrexate, mitroxatrone, gemcitabine, 5-fluorouracil (5-FU), bleomycin, doxorubicin, daunorubicin, epirubicin, idarubicin, valrubicin, paclitaxel, cabazitaxel, docetaxel, topotecan, irinotecan, etoposide, carboplatin, cisplatin, bortezomib, vinblastine, vincristine, vindesine, vinorelbine, diaziquone, mechlorethamine, mitomycin C, fludarabine, cytosine arabinoside, and combinations thereof.

13. 13. The pharmaceutical composition of any one of claims 1 to 12, wherein the cancer is selected from the group consisting of triple-negative breast cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, classical Hodgkin's lymphoma (cHL), bladder cancer, primary mediastinal B-cell lymphoma (PBMCL), urothelial carcinoma, microsatellite instability-high (MSI-H) solid tumor, mismatch repair-deficient (dMMR) solid tumor, gastric or gastroesophageal junction (GEJ) adenocarcinoma, esophageal squamous cell carcinoma, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, ovarian cancer, anal canal cancer, colorectal cancer, and melanoma.

14. 14. The pharmaceutical composition of claim 13, wherein the cancer is triple-negative breast cancer and the ICD-guided chemotherapy comprises gemcitabine and carboplatin.

15. 1. For use in a method for triple negative breast cancer therapy for a patient or patient population selected for extended progression free survival or overall survival, comprising administering to a subject the compound of claim 1, wherein the compound has the following structure: 【Chemistry 2】 or a pharmaceutically acceptable salt thereof, The method comprises the steps of: (i) determining whether the patient's triple-negative breast cancer has a surrounding microenvironment that is favorable for immune modulation; (ii) determining whether the patient is amenable to an immunogenic cell death (ICD)-inducing chemotherapy regimen comprising gemcitabine and carboplatin; (iii) if it is determined that the patient's triple-negative breast cancer is favorable to immunomodulation and the patient can be administered the ICD-guided chemotherapy regimen; (a) administering to said patient an effective amount of said CDK4 / 6 inhibitor; (b) administering to said patient an effective amount of gemcitabine, and (c) administering to said patient an effective amount of carboplatin. comprising the CDK4 / 6 inhibitor is administered prior to administration of gemcitabine and / or carboplatin; The pharmaceutical composition, wherein administration of the CDK4 / 6 inhibitor and the ICD-inducing chemotherapy regimen to the patient results in an extension of progression-free survival or overall survival compared to a patient with triple-negative breast cancer who receives ICD-inducing chemotherapy alone without a CDK4 / 6 inhibitor.

16. The pharmaceutical composition according to claim 15, wherein the triple-negative breast cancer is PD-L1 positive.

17. 17. The pharmaceutical composition of claim 15 or 16, wherein the CDK4 / 6 inhibitor is administered within about 24 hours prior to administration of gemcitabine and / or carboplatin.

18. 17. The pharmaceutical composition of claim 15 or 16, wherein the CDK4 / 6 inhibitor is administered within about 4 hours before administration of gemcitabine and / or carboplatin.

19. 17. The pharmaceutical composition of claim 15 or 16, wherein the CDK4 / 6 inhibitor is administered within about 30 minutes before administration of gemcitabine and / or carboplatin.

20. 2. A method for cancer therapy for a patient or patient population selected for extended progression free survival or overall survival comprising administering to said patient a compound having the following structure: 【Transformation 3】 or a pharmaceutically acceptable salt thereof, The method comprises the steps of: (i) determining whether the cancer has an elevated IFN-γ signature; (ii) determining whether the patient is amenable to an immunogenic cell death (ICD)-inducing chemotherapy regimen; (iii) When the patient's cancer has a high IFN-γ signature and the patient is determined to be able to administer the ICD-guided chemotherapy regimen, (a) administering an effective amount of said CDK4 / 6 inhibitor; (b) administering an effective amount of said ICD-guided chemotherapy regimen. comprising the CDK4 / 6 inhibitor is administered prior to administration of the ICD-inducing chemotherapy regimen; The pharmaceutical composition, wherein administration of the CDK4 / 6 inhibitor and the ICD-inducing chemotherapy regimen to the patient results in an extension of progression-free survival or overall survival compared to a patient with cancer who has been administered ICD-inducing chemotherapy alone without a CDK4 / 6 inhibitor.

21. 21. The pharmaceutical composition of claim 20, wherein the determination of whether the cancer has a high IFN-γ signature is based on the expression levels of the genes IDO1, CXCL10, CSCL9, HLA-DRA, STAT1, and IFN-γ in the tumor microenvironment.

22. 21. The pharmaceutical composition of claim 20, wherein the determination of whether the cancer has a high IFN-γ signature is based on a high Ayers et al. IFN-γ signature score.

Images