TWI835909B - Inhibitors of cyclin-dependent kinase 7 (cdk7) - Google Patents

Abstract

The present invention provides various compositions, including compounds of Formula (I) or (Ia), or a species thereof, and pharmaceutically acceptable salts, solvates (e.g. , hydrates), stereoisomer, tautomers, and isotopic forms thereof. Also provided are methods (or uses) and kits involving the compounds or pharmaceutically acceptable formulations for treating or preventing a disease (e.g. , a proliferative disease) in a subject. Administration of a compound or pharmaceutical composition described herein may inhibit the aberrant activity of cyclin-dependent kinase 7 (CDK7), and therefore, induce cellular apoptosis in the subject.

Description

Cyclin-dependent kinase 7 (CDK7) inhibitors

Members of the cyclin-dependent kinase (CDK) family are believed to play an important role in regulating cell proliferation. The primary target of the inhibitors described herein, CDK7, exists as a heterotrimeric complex in the cytosol and also forms the kinase core of the RNA polymerase (RNAP) II general transcription factor complex in the nucleus. Within this complex, CDK7 phosphorylates the C-terminal domain (CTD) of RNAP II, an essential step for initiating gene transcription. Disruptive RNAP II CTD phosphorylation has been shown to preferentially affect proteins with short half-lives, including proteins of the anti-apoptotic BCL-2 family. Since cancer cells have been shown to have the ability to circumvent cell death by upregulating BCL-2 family members, there is a mechanistic rationale for inhibiting cell proliferation by inhibiting CDK7. However, developing inhibitors selective for CDK7 is difficult, in part because the kinase domains of CDK family members share similar sequences and structural motifs.

The present invention provides selective CDK7 inhibitors, which are compounds having the formula disclosed herein (e.g., formula (I) or its subgenus or species) and in various embodiments, pharmaceutically acceptable salts, solvates (e.g., hydrates), stereoisomers or mixtures thereof, tautomers and isotopic forms (e.g., deuterated forms), wherein the various components of formula (I) (e.g., elements R ¹ , R ² , R ³ and R ⁴ of formula (I) and their subvariants) are as described herein. The compounds of the present invention exhibit surprising and unexpected superiority over other comparative compounds (in terms of selectivity for CDK7 over each of CDK2, CDK9 and CDK12); affinity for CDK7/cyclin H complex; and antiproliferative activity in a cell line model of triple-negative breast cancer. In addition, the compounds of the invention exhibit good bioavailability in a rat model. Thus, in a first embodiment, the invention provides a compound of a formula described herein (e.g., formula (I) or (Ia)) or a species thereof; in a second embodiment, the invention provides a salt thereof; in a third embodiment, the invention provides a solvate thereof; in a fourth embodiment, the invention provides a stereoisomer thereof; in a fifth embodiment, the invention provides a tautomer thereof; and in a sixth embodiment, the invention provides an isotopic form thereof. Salts, solvates, stereoisomers, tautomers or isotopic forms may be pharmaceutically acceptable and/or contained in pharmaceutically acceptable compositions, and in case of doubt, a given compound and its specified form may be a salt. For example, the present invention covers salts of the compound itself as well as salts of stereoisomers, tautomers, and isotopic forms of the compound. Similarly and in case of doubt, a given compound and a specified form thereof may be a solvate or an isotopic form. For example, the present invention covers solvates of the compound itself as well as solvates of salts, stereoisomers, tautomers, or isotopic forms thereof; the present invention covers isotopic forms of the compound itself as well as isotopic forms of salts, solvates, stereoisomers, or tautomers thereof. Such compositions (e.g., compounds, salts of stereoisomers, tautomers, or isotopic forms; compounds, solvates of stereoisomers, tautomers, or isotopic forms thereof; salts, solvates, stereoisomers, or isotopic forms of isomeric forms of compounds, etc.) constitute compositions of the present invention suitable for use as described herein.

In one embodiment, the invention features a compound of formula (I): , or a pharmaceutically acceptable salt thereof, wherein R ¹ is methyl or ethyl; R ² is methyl or ethyl; R ³ is 5-methylpiperidin-3-yl, 5,5-dimethyl piperidin-3-yl, 6-methylpiperidin-3-yl or 6,6-dimethylpiperidin-3-yl, wherein one or more hydrogen atoms in R ³ are optionally replaced by deuterium; and R ⁴ is -CF ₃ or chlorine. In certain embodiments, the compound has structural formula (Ia): , or a pharmaceutically acceptable salt thereof, where R ³ is . For example, the compound can be: , or may be a pharmaceutically acceptable salt of any of the foregoing compounds. In one embodiment, the compound is , or its pharmaceutically acceptable salt. In various embodiments, the compound is as just described; the compound is a pharmaceutically acceptable salt of the compound as described; the compound is as described except for one or more hydrogen atoms (e.g., one of ^R or more hydrogen atoms) substituted with deuterium; or the compound is a pharmaceutically acceptable salt of the illustrated compound in which one or more hydrogen atoms (eg, one or more hydrogen atoms in R ³ ) are substituted with deuterium.

In another embodiment, the invention features solvates (eg, hydrates) of compounds of formula (I). In other embodiments, the invention features solvates of pharmaceutically acceptable salts (e.g., hydrates) of compounds of Formula (I); solvates of tautomers (e.g., hydrates) of compounds of Formula (I) Hydrates); and solvates (such as hydrates) of isomeric forms of compounds of formula (I).

In another aspect, the invention features a pharmaceutical composition, which we may also refer to as a pharmaceutical formulation, comprising a compound as described above or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable salt thereof. Acceptable carrier. In other embodiments, pharmaceutical formulations include compounds of Formula (I) or (Ia), or solvates (eg, hydrates), stereoisomers, tautomers or isomeric forms of species thereof. The pharmaceutical compositions of the present invention may be formulated for oral administration and/or formulated into unit dosage forms.

In another aspect, the invention features methods or "uses" of the pharmaceutical compositions described herein for treating or preventing a disease, wherein the disease is a proliferative disease, an inflammatory disease, an autoinflammatory disease, an autoimmune disease, or an infectious disease in an individual in need thereof. The method of treatment may include the step of administering the pharmaceutical composition and the "uses" of the compositions of the invention may be used to prepare a medicament. In one embodiment, the disease is a proliferative disease (e.g., cancer, benign tumor, or pathological angiogenesis). In one embodiment, the cancer is a blood cancer. In one embodiment, the cancer is characterized by the presence of a solid tumor. In one embodiment, the various methods and uses described herein (e.g., for treating or preventing a proliferative disease, an inflammatory disease, an autoinflammatory disease, an autoimmune disease, or an infectious disease) are applied to an individual who has been determined to have one or more of the following (e.g., as determined in a biological sample of diseased cells obtained from the individual): elevated CDK7 expression or activity; high-grade cancer; a cellular phenotype in which steroid receptors are present and/or overexpressed; triple-negative breast cancer; and/or resistance to previously administered chemotherapeutic agents (e.g., CDK inhibitors such as palbociclib, or steroid receptor degraders such as palbociclib).

In another aspect, the invention features a kit comprising a pharmaceutical composition/formulation as described herein and instructions for use, and optionally a second agent selected from an antiproliferative agent, an anticancer agent, an immunosuppressant, and an analgesic. The kit may comprise a container having a compound described herein or a pharmaceutically acceptable salt, solvate, stereoisomer, tautomer, or isotopic form thereof, or a composition (e.g., a pharmaceutically acceptable composition or formulation as described herein) comprising a compound, a salt, solvate, stereoisomer, tautomer, or isotopic form thereof. In any embodiment of the kit, instructions for use may be included.

The pharmaceutically acceptable compositions of the present invention include compositions of the present invention (e.g., compounds described herein or pharmaceutically acceptable salts, solvates (e.g., hydrates), stereoisomers, tautomers, or isotopic forms thereof) and pharmaceutically acceptable carriers. In certain embodiments, the pharmaceutical composition includes a therapeutically or prophylactically effective amount of a compound of Formula I or a subgenus or species thereof, or a pharmaceutically acceptable salt, solvate, stereoisomer, tautomer, or isotopic form thereof. The pharmaceutical composition may be suitable for treating and/or preventing proliferative or infectious diseases, as further described below. The present invention also provides methods for making and using the compounds and other compositions described herein as therapeutic agents for preventing and/or treating diseases associated with overexpression and/or abnormal activity of CDK7. Diseases which may be mentioned include proliferative diseases (for example cancer (eg leukemia, melanoma, multiple myeloma), benign tumors and pathological angiogenesis), inflammatory diseases, autoinflammatory diseases, autoimmune diseases and infectious diseases.

Details of various embodiments of the invention are set forth herein. Other features, objects and advantages of the present invention will be apparent to those skilled in the art based on the detailed description, drawings, examples and patent scope of the present invention.

Cross-references to related applications

This application claims the benefit of U.S. Provisional Application No. 62/754,398, filed on November 1, 2018; U.S. Provisional Application No. 62/877,189, filed on July 22, 2019; and U.S. Provisional Application No. 62/915,983, filed on October 16, 2019. The contents of each of these prior applications are hereby incorporated by reference in their entirety.

Unless the context clearly indicates otherwise, the following definitions apply to the compositions and methods described herein. The definitions provided below apply to the language and grammatical variants of the defined terms (e.g., singular and plural forms of the terms), and some language variants are specifically mentioned below (e.g., "administration" vs. "administering" and "biologically active" vs. "biological activity"). Chemical elements are specified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th edition. In addition, the general principles of organic chemistry are well established, and those generally skilled in the art can consult, if necessary, Thomas Sorrell, Organic Chemistry , University Science Books, Sausalito, 1999; Smith and March, March 's Advanced Organic Chemistry , 5th edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations , VCH Publishers, Inc., New York, 1989; and Carruthers, Some Modern Methods of Organic Synthesis , 3rd edition, Cambridge University Press, Cambridge, 1987.

The term "about," when used in reference to a value, means any value or range of values within plus or minus 10% of the stated value (e.g., within plus or minus 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% of the stated value). For example, a dose of about 10 mg means any dose as low as 10% of 10 mg (9 mg) that is less than 10%, as high as 10% of 10 mg (11 mg) that is greater than 10%, and any dose or range of doses therebetween (e.g., 9-11 mg; 9.1-10.9 mg; 9.2-10.8 mg, etc.). In the case where the stated value (e.g., 100%) cannot be exceeded, "about" means any value or range of values up to and including 10% lower than the stated value (e.g., about 100% purity means 90%-100% pure (e.g., 95%-100% pure, 96%-100% pure, 97%-100% pure, etc.)). In the case where the instrument or technique for measuring the value has an error margin greater than 10%, the given value will be approximately the same as the stated value, with the proviso that the given value and the stated value are both within the error margin for the instrument or technique.

The term "administration" and variations thereof, such as "administering", generally refers to administering a compound described herein, a pharmaceutically acceptable salt, solvate, stereoisomer, tautomer or isotopic form thereof, or a composition (e.g., a pharmaceutical composition) containing one or more of the compound or its specified form to a subject (e.g., a human patient) or a system (e.g., a cell- or tissue-based system maintained in vitro); as a result of administration, the compound, its specified form, or a composition (e.g., a pharmaceutically acceptable salt) containing the compound or the specified form is introduced into the subject or system. In addition to the compositions of the invention, items may be "administered" that serve as positive controls, negative controls, placebos, or "second" agents used in combination therapy with the compositions of the invention, any of which may also be compounds. One of ordinary skill in the art will appreciate the variety of routes that may be used for administration to an individual or system, as appropriate. For example, the route of administration may be oral (i.e., by swallowing the pharmaceutical composition) or may be parenteral. More specifically, the route of administration can be bronchial (e.g., by bronchial instillation), oral (i.e., peroral), dermally (which can be or include topical application to the skin or intradermal, interdermal, or transdermal administration), intragastricly or enterally (i.e., directly applied to the stomach or intestines, respectively), intramedullary, intramuscularly, intranasally, intraperitoneally, intrathecally, intratumorally, intravenously (or intraarterially), intraventricularly, by application to or injection into a specific organ (e.g., intrahepatic), transmucosal (e.g., intrabuccal, rectal, sublingual, or vaginally), subcutaneously, tracheally (e.g., by intratracheal instillation), or ocular (e.g., topically, subconjunctivally, or intravitreally). Administration may involve intermittent dosing (i.e., by spacing doses at different times) and/or periodic dosing (i.e., by spacing doses at a common time period (e.g., hourly, daily, weekly, twice weekly, etc.)). In other embodiments, administration may involve continuous dosing (e.g., perfusion) over a selected time period (e.g., 1-2 hours).

The term "angiogenesis" refers to the formation and growth of new blood vessels. Normal angiogenesis occurs in healthy individuals during development and in the context of wound healing. However, patients with many different disease conditions, including cancer, diabetes (particularly the development of blindness associated with diabetes), age-related macular degeneration, rheumatoid arthritis, and psoriasis, experience excessive and harmful angiogenesis. Angiogenesis is undesirable when it creates blood vessels that support diseased cells (e.g., tumor cells), damage normal tissue (e.g., tissue within the eye), or promote tumor metastasis. We may refer to angiogenesis that accompanies and/or promotes disease conditions as "pathological angiogenesis."

Two events, two entities, or an event and an entity are "associated" with one another if one or more characteristics of the first (e.g., its presence, extent, and/or form) are associated with characteristics of the second. For example, a first entity (e.g., an enzyme (e.g., CDK7)), gene expression profile, gene signature (i.e., the gene expression pattern of a single set or combination of sets of genes in a cell has unique characteristics), metabolite, or event (e.g., bone marrow infiltration)) is associated with an event (e.g., the onset or development of a particular disease) if its presence, extent, and/or form is associated with the incidence, severity, and/or susceptibility of the disease (cancer disclosed herein). Correlation is often assessed in related groups. Two or more entities are physically "associated" with each other if they interact directly or indirectly such that they are in physical proximity to each other and/or remain in physical proximity to each other in a given situation (e.g., within a cell maintained under physiological conditions (e.g., within a cell culture) or within a pharmaceutical composition). Entities that are physically associated with each other may be covalently linked to each other or non-covalently associated by, for example, hydrogen bonds, van der Waals forces, hydrophobic interactions, magnetism, or a combination thereof. A compound described herein, or a salt, solvate, stereoisomer, tautomer, or isotopic form thereof, may non-covalently associate with CDK7.

The term "autoimmune disease" refers to a disease caused by an inappropriate immune response to substances and tissues normally found in the body. More simply put, an individual's immune system mistakes parts of the body for pathogens and attacks its own cells. Attack may be limited to certain organs (eg, in autoimmune thyroiditis) or may involve specific tissues in different locations (eg, Goodpasture's disease affects the basement membrane in both the lungs and kidneys). Autoimmune diseases include, but are not limited to, ANCA-associated vasculitis (e.g., Wegener's granulomatosis, microscopic polyangiitis), ankylosing spondylitis, antiphospholipid antibody syndrome, autoimmune thyroiditis, cardiomyopathy , Crohn's disease, dermatomyositis or polymyositis, glomerulonephritis, Cubast disease, Guillain-Barré disease, Hashimoto's thyroiditis, Lyme Lyme arthritis, lymphadenitis, necrotizing vasculitis, nodular periarteritis, psoriasis, pemphigus vulgaris, psoriatic arthritis, Reiter's syndrome, rheumatoid arthritis, scleroderma disease, Sjogren's syndrome, systemic lupus erythematosus (SLE), systemic sclerosis, ulcerative colitis and uveitis. Patients with some autoimmune diseases also experience significant inflammation (eg, rheumatoid arthritis and SLE), and therefore some diseases may be appropriately termed autoimmune or inflammatory diseases.

"Autoinflammatory diseases," also called "periodic fever syndromes," are diseases characterized by recurrent (intermittent) fevers and signs of systemic inflammation on blood tests (see Ciccarelli et al., Curr. Med. Chem. 21 (3): 261-269, 2013). The inflammatory episodes can be severe and accompanied by rashes or joint swelling, and patients are at risk for amyloidosis, a potentially fatal accumulation of blood proteins in vital organs. These diseases are distinct from autoimmune diseases and do not involve autoreactive T-lymphocytes or autoantibodies. These include, but are not limited to, Behcet's disease, Blau's syndrome, chronic recurrent multifocal osteomyelitis (CRMO), SAPHO syndrome, cryopyrin-associated periodic syndromes (CAPS), interleukin-1 receptor antagonist deficiency (DIRA), familial Mediterranean fever (FMF), NLRP12-related autoinflammatory disorders, neonatal-onset multisystem inflammatory disease (NOMID), Majeed syndrome, and Syndrome, periodic fever associated with mevalonate kinase deficiency (hyperimmunoglobulin D syndrome), periodic fever, aphthous stomatitis, pharyngitis, and adenopathy syndrome (PFAPA), purulent arthritis, pyoderma gangrenosum, acne (PAPA) syndrome, Schnitzer's syndrome, Sweet's syndrome, systemic juvenile idiopathic arthritis (sJIA) or Still's disease and adult-onset Still's disease (AOSD), and tumor necrosis factor (TNF) receptor-associated periodic syndrome (TRAPS).

The term "binding" and its variants (such as "bound" and "bind(s)"), particularly when used to refer to two or more entities, refers to the covalent or non-covalent association of the entities (e.g., a compound and an agent within a pharmaceutical composition or a compound and its target (e.g., CDK7) within a cell). "Direct" binding occurs when the two entities are in physical contact with each other (e.g., via a covalent or non-covalent chemical bond), while indirect binding occurs when at least one of the entities is in physical contact with an intervening entity that brings the entities into physical proximity with each other (e.g., within a complex). Binding can be assessed in a variety of contexts, such as in assays of completely or partially isolated entities or in more complex naturally occurring or model systems, such as in vivo or ex vivo tissues, organs, or cells. Analysis of binding can assess biological activity (eg, the ability of a compound described herein to inhibit the biological activity of a target (eg, CDK7)).

The term "biological sample" refers to a sample obtained from or derived from a biological source of interest (eg, tissue or organism (eg, animal or human patient) or cell culture). Biological samples may contain biological cells, tissues, or body fluids, or any combination thereof. For example, a biological sample may be or include ascites; blood; blood cells; body fluids, any of which may or may not include cells; bone marrow or components thereof (eg, hematopoietic cells, bone marrow adipose tissue, or stromal cells); brain Spinal fluid (CSF); feces; flexible fluid; free floating nucleic acids; gynecological fluids; immune infiltrates; lymph; peritoneal fluid; plasma; saliva; sputum; surgically obtained samples; samples from the skin or mucous membranes (e.g., in the nose, mouth, or vaginal) scraped or smeared tissue; tissue or fine-needle biopsy samples; urine; wash or laser, such as catheter lavage or bronchoalveolar lavage; or other body fluids, secretions, tissue and/or excreta. Biological samples may include cancer cells or immune cells, such as NK cells and/or macrophages, found in many tissues and organs, including the spleen and lymph nodes. The cells (eg, cancer cells) in the sample may have been obtained from the individual for whom treatment is intended. A sample used in the form in which it was obtained may be referred to as a "primary" sample, and a sample that has been further manipulated (for example, by adding or removing one or more components of the sample) may be referred to as "secondary" or "processed" Process the sample. Such processed samples may contain or be enriched for specific cell types (e.g., cells expressing CDK7, such as tumor cells), cellular components (e.g., membrane fractions), or cellular material (e.g., one that may encode CDK7 and may be amplified) or a variety of cellular proteins, including CDK7, DNA, or RNA (e.g., mRNA)).

The term "biologically active" describes an agent (e.g., described herein) that produces an observable biological effect or produces a biological system or model thereof (e.g., in humans, other animals, or systems maintained in cell/tissue culture or in vitro). compound). The "biological activity" of such an agent may be manifested by binding of the agent to a target (e.g., a cyclin-dependent kinase (e.g., CDK7)) and causing a biological pathway, event, or state (e.g., a disease condition) regulation (e.g., induction, enhancement, or inhibition). For example, the agent may modulate cellular activity (e.g., stimulate an immune response or inhibit homologous recombination repair), the time spent in a cell cycle phase (which may alter the rate of cell proliferation), or initiate an alternative pathway leading to cell death. Apoptosis or activation (which can cause tumor regression). Biological activity, and optionally its extent, can be assessed using known methods to detect the activity of any given intermediate or downstream product or any activity-related event (eg, inhibition of cell growth or tumor regression).

The term "cancer" refers to a disease in which biological cells exhibit an abnormal growth phenotype, characterized by uncontrolled cell proliferation to a degree that is detrimental to patients suffering from the disease. Cancers can be classified by the type of tissue in which they originate (histological type) and/or by the main location in the body where the cancer first appears. Cancers are generally grouped into six major categories based on histologic type: carcinoma; sarcoma; myeloma; leukemia; lymphoma; and mixed types. Cancer treated as described herein can be of any of these types and can include precancerous (eg, benign), malignant, pre-metastatic, metastatic, and/or non-metastatic cells. Patients with malignant tumors or malignant lesions have cancer. The present invention specifically identifies certain cancers to which its teachings may be particularly relevant, and one or more of these cancers may be characterized by solid tumors or by hematological tumors, which may also be referred to as hematological cancers (e.g., as described herein). type described). Although not all cancers manifest as solid tumors, we use the terms "cancer cell" and "tumor cell" interchangeably to refer to any malignant cell.

The term "carrier" refers to a diluent, adjuvant, excipient or other vehicle with which an active pharmaceutical agent (e.g., a compound of the present invention or a pharmaceutically acceptable salt, solvate, stereoisomer, tautomer or isotopic form thereof) is formulated for administration. The carrier in the amount and manner incorporated into the pharmaceutical composition will be non-toxic to the subject and will not destroy the biological activity of the active ingredient (e.g., compound or other specified form of the compound) formulated with it. The carrier can be a sterile or sterilizable liquid, such as water (e.g., water for injection) or a natural or synthetic oil (e.g., petroleum-based oil or mineral oil, animal oil or vegetable oil (e.g., peanut oil, soybean oil, sesame oil or rapeseed oil)). The carrier may also be a solid; a liquid comprising one or more solid components (eg, a salt, such as "normal saline"); a solid mixture; or a liquid mixture.

The term "comparable" means that two or more items (e.g., agents, entities, situations, setting conditions, etc.) are not consistent with each other, but are sufficiently similar to enable comparison between them so that a person of ordinary skill in the art will understand Yes, conclusions can be reasonably drawn based on observed differences or similarities. In some embodiments, comparable groups of conditions, situations, individuals, or groups are characterized by a plurality of substantially identical characteristics and one or a few different characteristics. One of ordinary skill in the art will understand what degree of consistency is required for two or more items to be considered comparable, within the context, and in any given situation. For example, two projects are comparable to each other when they share a sufficient number and type of substantially consistent characteristics to warrant the reasonable conclusion that any differences in the results obtained or observed phenomena from the projects are due to their different Caused or indicated by differences in characteristics. In some embodiments, comparable items serve as "controls." For example, a "control individual/population" may be an untreated (or placebo-untreated) individual/population suffering from the same disease as the treated individual/population.

The term "combination therapy" refers to situations in which an individual is exposed to two or more treatment regimens (eg, two or more therapeutic agents) to treat a single disease (eg, cancer). Two or more regimens may be administered simultaneously or sequentially (eg, all administrations of a first regimen are administered before any dose of a second regimen is administered by the same or different route of administration). For clarity, combination therapy does not require that the individual agents be administered together (or even necessarily simultaneously) in a single composition, but in some embodiments, two or more agents (e.g., compounds described herein and/or their designated forms) may be administered within the same time period (e.g., within the same day, week, or month), and may not be administered in combination, but in single compositions or even as combined compounds (e.g., associated or Combine to form a single chemical complex or entity (e.g., a covalent entity)).

The term "compound" means a chemical compound (eg, a compound represented by a structural formula depicted herein, a subgenus thereof, or a species thereof). Any given compound described herein may be biologically active (e.g., as a CDK7 inhibitor) and may be used for the purposes described herein, including therapeutic and prophylactic uses (e.g., when administered in a therapeutically or prophylactically effective amount when included in a pharmaceutical composition), used in a biological assay (eg, a diagnostic assay), administered to a patient, incorporated into a medicament or kit, or otherwise used as described herein). Two compounds with the same molecular formula but different arrangements of their atoms in space are called "stereoisomers". Stereoisomers of any structure mentioned or depicted may be enantiomers that are non-superimposable mirror images of each other, or are not mirror images of each other (e.g., cis / trans isomers and conformational isomers) of diastereomers. Such isomers include the R and S configurations of each asymmetric center, the Z and E double bond isomers, and the Z and E conformational isomers. Compositions containing individual types of stereochemical isomers as well as enantiomeric, diastereomeric and geometric (or conformational) mixtures of the compounds of the invention are within the scope of the invention. Enantiomers may be characterized by the absolute configuration of their asymmetric centers and described by the R-ordering and S-ordering rules of Cahn and Prelog, or by the rotation of the molecule about the plane of polarized light and designated as Dextral or levorotatory (i.e., the (+) or (-) isomer, respectively). Chiral compounds may exist as individual enantiomers or as mixtures thereof. A mixture containing equal proportions of enantiomers is called a "racemic mixture".

The terms "dosage form", "formulation", and "preparation" refer to a composition containing at least one compound of the invention, its salt, solvate, stereoisomer, tautomer or isotopic form, or other biologically or therapeutically active ingredients as described herein (e.g., a second agent suitable for combination therapy with a composition of the invention (i.e., the "first" agent/ingredient). The term "unit dosage form" refers to physically discrete units of a compound, its salt, solvate, stereoisomer, tautomer or isotopic form, any of which may be pharmaceutically acceptable, or formulated for administration to another biologically or therapeutically active ingredient in a subject. Typically, each such unit contains a predetermined amount of active ingredient, which may be an amount specified for a single dose (i.e., an amount expected to correlate with a desired outcome when administered as part of a therapeutic or preventive regimen) or a fraction thereof. One of ordinary skill in the art will appreciate that the total amount of a composition or medicament to be administered to a particular individual is determined by one or more attending physicians and may involve administration of multiple unit dosage forms (e.g., as described herein).

The term "dosage regimen" refers to one or more unit dosage forms administered to or prescribed for an individual, and typically includes more than one dose separated by a time period (e.g., as described herein). The multiple dosage forms administered within a dosing regimen can have the same unit dosage or different amounts. For example, a dosing regimen can include a first dose of a first dosage, followed by one or more additional doses of a second dosage that is the same or different than the first dosage.

An "effective amount" refers to an amount of an agent (eg, a compound described herein, whether or not the "second" agent of the invention) that is administered to produce the desired effect. In some embodiments, the term refers to an amount sufficient to treat a disease when administered according to a therapeutic dosing regimen to a population suffering from or susceptible to the disease, in which case the effective amount may also be referred to as a "therapeutically effective amount." Those skilled in the art should understand that a therapeutically effective amount may not achieve successful treatment in any particular individual. Rather, a therapeutically effective amount provides the desired pharmacological response in a substantial number or number of individuals when administered to a patient population in need of such treatment. Where an agent is administered prophylactically, a "prophylactically effective amount" provides the desired result (i.e., delays the onset of one or more signs or symptoms of the disease) in a significant or number of individuals in a population that does not exhibit Signs and/or symptoms of disease. References to an effective amount may refer to the dose of the drug administered or as measured in one or more specific tissues (e.g., tissue affected by disease) or fluid (e.g., blood, saliva, urine, etc.) following administration Quantity mentioned.

The term "hydrate" refers to a compound or a specified form thereof combined with water; as understood in the art, a hydrate is a solvent complex in which the solvent is water. The amount of water contained in a hydrate can be expressed as the ratio of the number of water molecules to the number of compound molecules. Thus, a hydrate of a compound can be represented by a general formula such as R· _xH2O , where R is the compound and x is a number greater than 0. For example, when x is 1, the hydrate is a monohydrate; when x is 0.5, the hydrate is a hemihydrate; when x is 2, the hydrate is a dihydrate; and when x is 6, the hydrate is a hexahydrate.

"Improve", "increase" or "reduce(s)/(decrease(s)" (and obvious variants such as "improved" or "improving") are terms used to describe the way in which a value changes or has changed relative to a reference value. For example, a measurement obtained from a patient (or a biological sample obtained therefrom) before treatment may increase or decrease/decrease relative to the measurement obtained in the same patient, a control patient, an average patient population, or a biological sample obtained therefrom during or after treatment. The value may be improved in either case, depending on whether the increase or decrease is associated with a positive treatment outcome.

As used herein, the term "inflammatory disease" refers to a disease caused by, resulting from, or causing inflammation, or a dysregulated inflammatory response that elicits an amplified response by macrophages, granulocytes, and/or T lymphocytes that Dysregulated inflammatory responses cause abnormal tissue damage and/or cell death. Inflammatory diseases may be acute or chronic and may arise from infectious agents or non-infectious causes. Inflammatory diseases include, but are not limited to, acute anaphylaxis, adult respiratory distress syndrome (ARDS), allergies, allograft rejection, ankylosing spondylitis, appendicitis, arteriosclerosis, arteritis (e.g., giant cell arteritis), asbestosis disease, atherosclerosis, asthma, arthritis (such as gouty arthritis, degenerative arthritis, inflammatory arthritis and rheumatoid arthritis), osteoarthritis, autoimmune diseases, beryllium toxicity, blepharitis, Bronchiectasis, bronchiolitis, bronchitis (e.g., chronic bronchitis), bursitis, interstitial pneumonia, cellulitis, cervicitis, cholangitis, chorioamnionitis, conjunctivitis, chronic cholecystitis, Crohn's disease disease, cystic fibrosis, cystitis, dacryoadenitis, delayed allergic reactions (e.g., poison ivy dermatitis), dermatomyositis, exfoliative interstitial pneumonia, diabetes (e.g., type I), encephalitis, Endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, adhesions, allergic alveolitis, fasciitis, fibrositis, gastritis, gastroenteritis, giant cell interstitium Pneumonia, gingivitis, glomerulonephritis, Cubase disease, Graves' disease, Hashimoto's thyroiditis, hay fever, hepatitis, host versus graft rejection, ileitis, immediate anaphylaxis , inflammatory bowel disease (IBD), inflammatory skin disease, iritis, ischemia (ischemic injury), laryngitis, lymphoid interstitial pneumonia, mixed connective tissue disease, multiple sclerosis, Myasthenia gravis, osteomyelitis, myocarditis, necrotizing enterocolitis, nephritis, umbilical corditis, oophoritis, testicularitis, osteitis, osteomyelitis, optic neuritis, otitis media, pancreatitis, mumps, pemphigoid, pemphigus , pericarditis, pernicious anemia, pharyngitis, phlebitis, pleurisy, pneumoconiosis, pneumonia, local pneumonia, polymyalgia rheumatica (PMR), polymyositis, proctitis, progressive systemic sclerosing cholangitis, prostate inflammation, psoriasis, pyelonephritis, reperfusion injury, respiratory tract inflammation, rheumatic fever, rhinitis, salpingitis, sarcoidosis, scleroderma, silicosis, sinusitis, Sjogren's syndrome, oral synovitis, systemic lupus erythematosus (SLE), talcopulmonary disease, temporal arteritis, tendonitis, testicularitis, tonsillitis, transverse myelitis, necrotizing fasciitis, ulcerative colitis , urethritis, cystitis, common interstitial pneumonia (UIP), uveitis, vaginitis, vasculitis, vulvitis, labial vaginitis, and Wegener's granulomatosis and related forms of vasculitis (temporal arteritis and polyarteritis nodosa).

The term "inhibitor" refers to an agent, including a compound described herein or a specified form thereof, whose presence (e.g., at a certain level or in a certain form) is associated with a reduction in the expression or activity of another agent (i.e., the inhibitory agent or target) or a reduction in the occurrence of an event (e.g., cell proliferation, tumor development or metastasis, inflammation, infection, or autoimmunity). In some embodiments, the inhibitor exerts its effect on the target by binding directly or indirectly to the target, via covalent bonds or non-covalent associations. Inhibition can be assessed in vitro (e.g., in cells, tissues, or tissue cultures or systems) or in vivo (e.g., in patients or animal models) via computer simulations.

The term "isotopic form" is used to describe compounds containing at least one isotope substitution; the replacement of an isotope of an atom with another isotope of that atom. For example, the substitution may be ^2H (deuterium) or ^3H (tritium) instead of ^1H . Accordingly, we may use the term " ^1H ", "H", or "hydrogen atom" to refer to the naturally occurring form of hydrogen having a single proton in its nucleus. Other substitutions in the form of isotopes include ¹¹ C, ¹³ C or ¹⁴ C for ¹² C; ¹³ N or ¹⁵ N for ¹⁴ N; ¹⁷ O or ¹⁸ O for ¹⁶ O; ³⁶ Cl for ³⁵ C; ¹⁸ F for ¹⁹ F; ¹³¹ I replaces ¹²⁷ I; etc. Such compounds have been used, for example, as analytical tools, as probes in biological assays (eg, diagnostic assays), and/or as therapeutic or prophylactic agents for use according to the invention. Specifically, substitution of hydrogen with deuterium ( ^2H ) isotopes can slow metabolism, shift metabolism to other sites on the compound, slow racemization, and/or have a detrimental effect on the pharmacokinetics of the compound that may be beneficial (e.g., therapeutically beneficial). Other functions.

The terms "tumor" or "neoplasm" are used interchangeably herein and refer to an abnormal mass of tissue in which the growth of the mass exceeds and is uncoordinated with normal tissue growth. A tumor or neoplasm may be "benign" or "malignant," depending on the following characteristics: degree of cell differentiation (including morphology and function), growth rate, local invasion, and metastasis. "Benign tumors" are usually well differentiated, have a slower growth rate than malignant tumors, and remain localized to the primary site (i.e., do not have the ability to infiltrate, invade, or metastasize to distant sites). Benign tumors include, but are not limited to, verrucae, adenomas, chondromas, intraepithelial tumors, lentigos, adenomas, sebaceous hyperplasia, seborrheic keratosis, and senile hemangiomas. Benign tumors may also be tuberous sclerosis or tuberous sclerosis complex (TSC) or epiloia (from "epilepsy, low intelligence, sebaceous adenoma"). Benign tumors may subsequently give rise to malignant tumors (thought to occur due to genetic changes in a subset of the tumor's malignant cells), and such tumors are called "premalignant tumors." An exemplary premalignant tumor is a teratoma. In contrast, "malignant tumors" are generally poorly differentiated (regressive) and grow rapidly with progressive infiltration, invasion, and destruction of surrounding tissues. Malignant tumors also generally have the ability to metastasize to distant sites.

The terms "patient" and "subject" refer to any organism to which a compound described herein or a specified form thereof is administered according to the present invention, e.g., for experimental, diagnostic, prophylactic and/or therapeutic purposes. Typical subjects include animals (e.g., mammals, such as mice, rats, rabbits, non-human primates and humans; insects; worms, etc.). In some embodiments, the subject suffers from a disease described herein (e.g., a proliferative disease, such as cancer).

A "pharmaceutical composition" or "pharmaceutically acceptable composition" (we may also refer to it as a "pharmaceutical formulation" or "pharmaceutically acceptable formulation") is a composition/formulation in which an active agent (e.g., an active pharmaceutical ingredient (e.g., a compound, salt, solvate, stereoisomer, tautomer or isotopic form thereof)) is formulated with one or more pharmaceutically acceptable carriers. The active agent/ingredient may be present in an amount suitable for a unit dose for administration in a therapeutic regimen that exhibits a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. Pharmaceutical compositions may be specifically formulated for administration in solid or liquid form, including forms prepared for oral or parenteral administration. For oral administration, the pharmaceutical composition can be formulated, for example, as an aqueous or non-aqueous solution or suspension or tablet or capsule. For systemic absorption through the oral cavity, the composition can be formulated for buccal administration, sublingual administration, or in a paste for application to the tongue. For parenteral administration, the composition can be formulated, for example, as a sterile solution or suspension for subcutaneous, intramuscular, intravenous, intraarterial, intraperitoneal, intratumoral, or epidural injection. The pharmaceutical composition comprising the active agent/ingredient (e.g., a compound described herein or a specified form thereof) can also be formulated as a sustained release formulation or as a cream, ointment, controlled release patch, or spray for topical application. Creams, ointments, foams, gels and plasters may also be applied to the mucous membranes of the nose, mouth, vagina and rectum. Any of the compounds described herein and any pharmaceutical compositions containing such compounds may also be referred to as "agents."

The term "pharmaceutically acceptable" when applied to a carrier used to formulate a composition disclosed herein (e.g., a pharmaceutical composition) means that the carrier is compatible with the other ingredients of the composition and is not deleterious to the patient (e.g., the carrier is nontoxic in the amounts required and/or administered (e.g., in a unit dosage form)).

The term "pharmaceutically acceptable", when applied to salts, solvates, stereoisomers, tautomers or isotopic forms of the compounds described herein, means, within the scope of sound medical judgment, suitable for Contact with tissues of humans (e.g., patients) and lower animals (including, but not limited to, mice and rats used in laboratory research) without unacceptable toxicity, irritation, allergic reactions and similar effects, and can be used with Use salts, solvates, stereoisomers, tautomers or isotopic forms in a manner that is reasonably proportional to the advantages and disadvantages. Many pharmaceutically acceptable salts are well known in the art ( see, eg, Berge et al., J. Pharm. Sci. 66 :1-19, 1977). Pharmaceutically unacceptable salts, solvates, stereoisomers, tautomers or isotopic forms of the compounds of the invention are also within the scope of the invention and are useful, for example, in chemical processes and synthesis and in vitro experiments. utility. In pharmaceutically unacceptable compositions, a compound, its salt, solvate, stereoisomer, tautomer or isotopic form may be present in an overly concentrated or overly diluted amount for administration to a patient.

A "polypeptide" is a polymer of amino acid residues, regardless of length, origin, or post-translational modifications; it includes, but is not limited to, full-length naturally occurring proteins. We may refer to a polypeptide as the "target" of a composition when it binds (e.g., specifically binds) or otherwise interacts with a composition described herein. When used in connection with the occurrence of a disease (e.g. a proliferative disease such as cancer), the terms "prevent(s)", "prevention", "prophylaxis/prophylactic" and the like mean Reduce the risk of contracting a disease and/or delay the onset of one or more of its signs or symptoms. Prevention can be considered complete when the onset has been delayed for a predetermined period of time.

"Proliferative diseases" are diseases characterized by excessive proliferation of cells. Proliferative diseases are associated with: (1) pathological proliferation of normally quiescent or normally proliferating cells; (2) pathological migration of cells from their normal location (e.g., neoplastic cell carcinoma metastasis); (3) proteolytic enzymes ( Pathological manifestations such as matrix metalloproteinases (e.g., collagenases, gelatinases, and elastases), which can cause unwanted turnover of cellular matrix; and/or (4) pathological angiogenesis, as in proliferative retinopathy and occurs in tumor metastasis. Exemplary proliferative diseases include cancer, benign neoplasms, and angiogenesis that accompany and contribute to the disease condition (defined above as pathological angiogenesis).

The term "reference" describes a standard or control to which a comparison is made. For example, an agent, animal, cell, individual, population, sample (e.g., biological sample), sequence, or value of interest is compared to a reference or control agent, animal, cell, individual, population, sample, sequence, or value. In some embodiments, a reference or control is physically tested while the agent of interest is tested and/or assayed. In other embodiments, a reference or control is a historical reference or control implemented in a tangible medium, as appropriate. Generally, as one of ordinary skill in the art would understand, reference or comparators are identified or characterized under comparable conditions to those in the evaluation, and one of ordinary skill in the art would understand when sufficient similarity exists to justify reliance on a particular possible reference or comparator and/or comparison results.

The term "response" with respect to treatment can refer to any beneficial change in a condition in an individual that occurs as a result of or is associated with treatment. Such changes can include stabilization of the condition (e.g., prevention of worsening that would occur in the absence of treatment), improvement in symptoms of the condition, and/or improvement in the prospects for cure of the condition, etc. The response can be a cellular response (e.g., a tumor response) and can be measured using a variety of criteria known in the art, including clinical criteria and target criteria. Techniques used to assess response include, but are not limited to, analytical assessment, clinical testing, positron emission tomography, X-ray, CT scan, MRI, ultrasound, endoscopy, laparoscopy, assessment of the presence or amount of tumor markers in a sample obtained from an individual, cytology and/or histology. Methods and guidelines for measuring tumor response, for assessing response to treatment are discussed in Therasse et al. ( J. Natl. Cancer Inst. , 92(3) :205-216, 2000). The exact response criteria may be selected in any appropriate manner by one of ordinary skill in the art, with the proviso that when comparing groups of cancers and/or patients, the groups being compared are evaluated based on the same or comparable criteria used to determine response rate.

The term "solvolyte" refers to a compound formed by the combination of molecules of a solute and molecules of a solvent. Solvents that can be used to form a solvate include water, methanol, ethanol, acetic acid, DMSO (dimethyl sulfoxide), THF (tetrahydrofuran), diethyl ether and similar solvents. Solvolytes in which the solvent is water are called hydrates. The compound can be prepared in the form of a liquid or solid solvate, such as a crystalline solvate. The solvate can be pharmaceutically acceptable and can be a stoichiometric or non-stoichiometric solvate. In certain circumstances, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate will be able to separate. "Solvent" encompasses both solution phase and isolatable solvents, and representative solvents include hydrates, ethanolates and methanolates.

As used herein with reference to an agent (e.g., a compound) having an activity (e.g., inhibiting a target), the term "specificity" means that the agent discriminates between potential target entities or conditions. For example, an agent "specifically" binds to its intended target (or otherwise specifically inhibits its target) if it better binds or otherwise inhibits the expression or activity of that target in the presence of one or more alternative targets. Although the invention is not limited in this regard, specific and direct interactions may depend on the recognition of specific structural features of the target (e.g., antigenic determinants, gaps, or binding sites). Specificity need not be absolute; the degree of specificity need only be sufficient to produce effective treatment without unacceptable side effects. The specificity of an agent can be assessed by comparing the effect of the agent on an intended target or condition relative to its effect on a different target or condition. The effects on the intended and different targets can be determined separately or the effect on the intended target can be determined and compared to a reference standard developed at an earlier time (e.g., a reference specific binder or a reference non-specific binder). In some embodiments, an agent undetectably binds to a competing alternative target under conditions where it detectably (and preferably significantly) binds to its intended target, and/or undetectably inhibits the expression or activity of a competing target under conditions where it detectably (and preferably significantly) inhibits the expression or activity of its intended target. The compounds of the invention or their salts, solvates, stereoisomers, tautomers or isotopic forms may exhibit a higher on-rate, lower off-rate, increased affinity, decreased dissociation rate and/or increased stability relative to their target compared to a competing surrogate target, and any of these parameters may be evaluated in the methods of the invention.

The term "substantially" refers to the qualitative condition of exhibiting the characteristic or characteristic of interest to an overall or nearly overall degree or level. Those skilled in the art will understand that biological and chemical phenomena rarely, if ever, proceed to completion and/or proceed to completeness or rarely achieve or avoid absolute results. The term "substantially" is therefore used herein to capture the underlying lack of completeness inherent in many biological and chemical phenomena. For example, a chemical reaction can be characterized as substantially complete, even if the yield is well below 100%. When certain characteristics are approximately the same and/or exhibit approximately the same activity, those characteristics may also be considered "substantially the same". For example, two nearly identical compounds that produce approximately the same effect on an event (eg, cell proliferation) may be described as substantially similar. With respect to the purity of a compound or composition, it is defined below as "substantially pure."

Individuals who are "vulnerable" to a disease (such as cancer) have a higher than average risk of developing the disease. In some embodiments, such individuals do not exhibit any disease symptoms. In some embodiments, such individuals have not yet been diagnosed with the disease. In some embodiments, such individuals have been exposed to conditions associated with developing a disease (eg, exposure to carcinogens). In some embodiments, the risk of developing a disease is a population-based risk.

"Reduction of symptoms or symptoms" occurs when the magnitude (e.g., intensity, severity, etc.) and/or frequency of one or more objective signs or subjective symptoms of a specific disease decreases. Delaying the onset of a specific sign or symptom is a form of reducing the frequency of that sign or symptom. Alleviation of signs or symptoms may be achieved, for example, by "therapeutically active" compounds.

When used to refer to a compound or designated form thereof described herein, the term "substantially pure" means that a preparation of the compound or designated form thereof is greater than about 85% (w/w) of the compound or designated form thereof (e.g. Greater than about 90%, 95%, 97%, 98%, 99% or 99.9% of the compound or salt, solvate, stereoisomer, tautomer or isomer).

"Tautomers" are interchangeable forms of a specific compound's structure that change by substitution of hydrogen atoms and electrons. Therefore, two chemical structures can be kept in equilibrium through the movement of pi electrons and atoms (usually H). For example, enols and ketones are tautomers because they can be rapidly converted into each other by treatment with acids or bases. Another example of tautomerism is the acidified and nitro forms of phenylnitromethane, also formed by acid or base treatment. Tautomeric forms can be used to optimize chemical reactivity and biological activity of compounds of interest.

"Treatment regimen" means a dosage regimen that is associated with a desired therapeutic outcome when administered in a relevant population.

The term "treatment" and its linguistic variations, such as "treat(s)" and "treating", refers to any use of a pharmaceutical composition or administration of a therapy that partially or substantially completely relieves, ameliorates, alleviates, inhibits, reduces the severity of, and/or reduces the incidence of, one or more signs or symptoms of a particular disease (e.g., a proliferative disease, such as cancer). The individual being treated (or identified as a candidate for treatment) may only display early signs or symptoms of the disease, or may display one or more established or advanced signs or symptoms of the disease. "Treatment" is distinguished from "prevention", which involves delaying the onset of one or more signs or symptoms of a disease. In that case, the individual will not exhibit signs and/or symptoms of the disease and/or may be known to have one or more susceptibility factors that are statistically associated with an increased risk of developing the relevant disease. However, once a patient exhibits signs or symptoms of the disease and has been treated, the treatment may continue to delay the progression of the disease (e.g., in the case of localized cancer, treatment may delay tumor development (i.e., growth) or cancer metastasis) or to delay or prevent recurrence (e.g., tumor recurrence). The applicants describe herein the compounds of the invention having structural formula (I): , or a pharmaceutically acceptable salt, solvent (e.g., hydrate), stereoisomer, tautomer or isotope thereof. In formula (I), R ¹ is methyl or ethyl; R ² is methyl or ethyl; R ³ is 5-methylpiperidin-3-yl, 5,5-dimethylpiperidin-3-yl, 6-methylpiperidin-3-yl or 6,6-dimethylpiperidin-3-yl; and R ⁴ is -CF ₃ or chlorine. In some embodiments, in a compound of formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer or tautomer thereof, one or more atoms (e.g., one or more carbon atoms and/or hydrogen atoms in, for example, ring structures R ¹ , R ² , R ³ and/or R ⁴ ) are isotopically replaced with the atoms originally present (e.g., ¹² C originally present is replaced with ¹³ C or ¹⁴ C and/or ¹ H originally present is replaced with ² H or ³ H). In some embodiments, the compound has formula (Ia): , or a pharmaceutically acceptable salt, solvate (e.g., hydrate), stereoisomer, tautomer or isotopic form (e.g., a pharmaceutically acceptable salt thereof). R ¹ , R ² and R ⁴ are as defined for formula (I), and R ³ is As indicated above with respect to formula (I), any compound of formula (Ia) or any salt, solvate, stereoisomer or tautomer thereof may be an isotopic variant (e.g., one or more carbon and/or hydrogen atoms in the ring structures R ¹ , R ² , R ³ and/or R ⁴ are replaced by their isotopes (e.g., deuterium replaces ¹ H)).

In some embodiments of the formulae disclosed herein (e.g., Formula (I) or (Ia)), each of ^R1 and ^R2 is independently methyl, _{-CD3 ,} ethyl, _{-CD2CD3 , -CH2CD3 ,} or _-CH2CD3 , wherein "D" represents _deuterium .

In some embodiments of structural formula (I) or formula (Ia), the compound is , or a pharmaceutically acceptable salt, solvate (eg, hydrate), stereoisomer or tautomer thereof, or an isomeric form of any of the foregoing. In some embodiments, the compound is: , or a pharmaceutically acceptable salt, solvate (e.g., hydrate), stereoisomer or tautomer (e.g., a pharmaceutically acceptable salt thereof) of an isotopic variant of any of the foregoing. ). Isotopic variants may be as otherwise described above (e.g., one or more hydrogen atoms (e.g., in a substituent (medium) deuterium replacement as appropriate). In some embodiments of structural formula (I) or formula (Ia), R ³ is (S)-5-methylpiperidin-3-yl, (S)-5,5-dimethylpiperidin-3 -yl, (S)-6-methylpiperidin-3-yl, 6,6-dimethylpiperidin-3-yl, (S)-5-trideuteromethylpiperidin-3-yl, 5,5-trideuteromethylpiperidin-3-yl, (S)-6-trideuteromethylpiperidin-3-yl or 6,6-di-trideuteromethylpiperidin-3-yl base.

In various independent embodiments of structural formula (I), (Ia) or species thereof: R ¹ is methyl or ethyl; R ² is methyl or ethyl; R ¹ and R ² are both methyl; or R ¹ and R ² are both ethyl groups. In various embodiments, ^R3 is (S)-5-methylpiperidin-3-yl, 5,5-dimethylpiperidin-3-yl, (S)-6-methylpiperidin-3 -yl or 6,6-dimethylpiperidin-3-yl; R ³ is (S)-5-methylpiperidin-3-yl; R ³ is 5,5-dimethylpiperidin-3- base; R ³ is (S)-6-methylpiperidin-3-yl; or R ³ is 6,6-dimethylpiperidin-3-yl.

In some embodiments of Structural Formula (I) or Formula (Ia), R ⁴ is -CF ₃ .

Pharmaceutically acceptable salts or solvates, stereoisomers, tautomers or isotopic forms of the compounds described herein include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable acid addition salts are amines with inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid) or organic acids (such as acetic acid, oxalic acid, maleic acid, tartaric acid) , citric acid, succinic acid or malonic acid) or salts formed by using other methods known in the art (such as ion exchange). Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, besylate, and hydrogen sulfate. Salt, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentane propionate, digluconate, lauryl sulfate, ethane sulfonate, formic acid Salt, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, enanthate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate , lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methane sulfonate, 2-naphthalene sulfonate, nicotinate, Nitrate, oleate, oxalate, palmitate, pamoate, pectate, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, Stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate and similar salts.

Salts of any of the compounds described herein can also be derived from appropriate bases, including alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium and the like. Where appropriate, other pharmaceutically acceptable salts include ammonium, quaternary ammonium and use of counter ions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and Aryl sulfonate) formed amine cation.

The compounds of the invention, or pharmaceutically acceptable salts, solvates, stereoisomers, tautomers or isotopic forms thereof, may have one or more of the following properties: (1) With respect to K _i , at The specificity for CDK7 in an enzymatic assay is at least or about 25 times greater (e.g., at least or about 50 times, 100 times, 200 times, 300 times, or 400 times) than each of CDK2, CDK9, and CDK12; (2) Specificity for CDK7 in an enzymatic assay is at least or about 200-fold greater (eg, at least or about 300-fold, 400-fold, or 500-fold) than for each of CDK2, CDK9, and CDK12 with respect to _IC50 ; (3 ) binding to the CDK7/cyclin H complex with a K _d of less than 150 pM (e.g., less than 120 pM, 110 pM, or 100 pM) as measured by surface plasmon resonance (SPR); and (4) in the presence of Use HCC70 cells with an _EC50 of less than 10 nM (e.g., less than 5 nM, 4 nM, 3 nM, 2 nM, or 1 nM) in proliferation assays. These properties make the compounds or their designated forms particularly suitable for therapies requiring strong and specific inhibition of CDK7 without concomitant inhibition of other CDKs, especially CDK2, CDK9 and CDK12.

Pharmaceutical compositions and kits: The present invention provides pharmaceutical compositions, which include compounds of formula I, subgenus or species thereof, or pharmaceutically acceptable salts, solvates (such as hydrates), stereoisomers thereof , tautomer or isotopic forms, and pharmaceutically acceptable carriers selected as appropriate. In certain embodiments, a pharmaceutical composition includes: a compound of Formula (I) or (Ia) or a species thereof or a pharmaceutically acceptable salt thereof; Formula (I) in the form of a solvate (e.g., hydrate) or (Ia) a compound or a species thereof; a compound of formula (I) or (Ia) or a species thereof or a mixture thereof in stereoisomeric form (for example, where the stereoisomers are enantiomers or racemic mixtures thereof ); a compound of formula (I) or (Ia) or a species thereof in tautomeric form; or any of the foregoing in isotopic form. As indicated, the pharmaceutical composition may include one or more pharmaceutically acceptable carriers, in which the active agent/ingredient may be provided in an effective amount (eg, a therapeutically effective amount or a prophylactically effective amount).

The pharmaceutical compositions described herein may be prepared by any method known in the pharmacological art. Generally, such preparation methods include the steps of bringing into contact a compound described herein, or a pharmaceutically acceptable salt, solvate, stereoisomer, tautomer or isotopic form thereof, with a carrier and/or One or more other active ingredients (e.g., a second agent as described herein) and/or accessory ingredients are associated and the product is subsequently shaped and/or packaged into the desired single- or multi-dose units as necessary and/or desired ( such as oral administration). Pharmaceutical compositions may be packaged in a variety of ways, including in bulk containers and as single unit doses (containing, for example, discrete predetermined amounts of an active agent) or plural forms thereof, and any such packaged or divided dosage forms are within the scope of the present invention. . The amount of active ingredient may be equal to that amount constituting a unit dose or a suitable fraction of the dose, such as one-half or one-third of the dose.

The relative amounts of the active agent/ingredient, one or more pharmaceutically acceptable carriers and/or any additional ingredients in the pharmaceutical compositions of the present invention may vary depending on the identity, size and/or condition of the individual being treated and further vary depending on the route of administration of the composition and the disease to be treated. By way of example, the composition may contain between about 0.1% and 99.9% (w/w or w/v) of the active agent/ingredient.

Pharmaceutically acceptable carriers suitable for use in the manufacture of pharmaceutical compositions described herein are well known in the pharmaceutical formulation art and include inert diluents, dispersing and/or granulating agents, surfactants and/or emulsifiers , disintegrants, binders, preservatives, buffers, lubricants and/or oils. Pharmaceutically acceptable carriers suitable for use in the manufacture of pharmaceutical compositions described herein include, but are not limited to, ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as phosphoric acid, glycine, sorbic acid, potassium sorbate), saturated vegetable fatty acids, water, salts or partial glyceride mixtures of electrolytes (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride , zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxy Propylene - block polymer, polyethylene glycol and lanolin.

The compositions of the present invention may be administered orally and oral formulations are within the scope of the present invention. Such orally acceptable dosage forms include, but are not limited to, capsules, tablets, suspensions and solutions (e.g., aqueous suspensions and solutions). In the case of tablets, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate may also be included. In the case of capsules, suitable diluents include lactose and dried corn starch. When preparing aqueous suspensions, the active agent/ingredient may be combined with an emulsifier and a suspending agent. In any oral formulation, sweeteners, flavorings or coloring agents may also be added. In any of the various embodiments described herein, oral formulations can be formulated for immediate release or sustained/delayed release. Provided compositions can also be in microencapsulated form.

Compositions suitable for buccal or sublingual administration include lozenges, buccal lozenges and tablets. Formulations may also be prepared for use in subcutaneous, intravenous, intramuscular, intraocular, intravitreal, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intraperitoneal intralesional and transcranial injection or infusion techniques. Preferably, the composition is administered orally, subcutaneously, intraperitoneally or intravenously. Sterile injectable forms of the compositions of the invention may be aqueous or oleaginous suspensions. Such suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents which may be used are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils are routinely used as solvents or suspending media.

Although the descriptions of pharmaceutical compositions provided herein are generally directed to pharmaceutical compositions suitable for administration to humans, those skilled in the art will understand that such compositions are generally suitable for administration to all types of animals. It is well understood that pharmaceutical compositions suitable for administration to humans may be modified to render the compositions suitable for administration to various animals, and that such modifications may only be designed and/or carried out by a skilled veterinary pharmacologist.

For ease of administration and uniformity of dosage, the compounds described herein are typically formulated in unit dosage forms (e.g., single unit dosage forms). However, it is understood that the total daily dosage of the compositions of the present invention will be determined by the attending physician within the scope of sound medical judgment. The specific therapeutically or prophylactically effective amount for any particular individual or organism will depend on a variety of factors, including the severity of the disease and condition being treated; the activity of the specific active ingredient employed; the specific composition employed; the age, weight, general health, sex, and diet of the individual; the time of administration, route of administration, and rate of excretion of the specific active ingredient employed; the duration of treatment; drugs used in combination or concomitantly with the specific active ingredient employed; and similar factors well known in the medical field.

The exact amount of compound required to achieve an effective amount may vary from individual to individual, depending, for example, on the species, age and general condition of the individual, the severity of the side effects, the disease to be treated, and the specific species or species to be administered. The identity of the compound, mode of administration and similar factors. The required dose can be delivered three times a day, twice a day, once a day, every other day, every three days, weekly, every two weeks, every three weeks, or every four weeks. In certain embodiments, the desired dose may be delivered using multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, Twelve, thirteen, fourteen or more throws).

In certain embodiments, an effective amount of the compound for administration to a 70 kg adult one or more times per day may comprise about 1-100 mg, about 1-50 mg, about 2-20 mg, or about 3-15 mg . And here, whenever a range is mentioned, the endpoints are included.

In certain embodiments, the compositions of the invention may be administered once daily. The dosage of the compound of formula I or its subgenus or species or a specified form thereof (e.g., a salt thereof) may be about 1-100 mg, about 1-50 mg, about 1-25 mg, about 2-20 mg, about 5-15 mg, about 10-15 mg, or about 13-14 mg.

In certain embodiments, compositions of the present invention may be administered twice daily. In some embodiments, the compound of Formula I, or a subgenus or species thereof, is used in a dosage of about 0.5 mg to about 50 mg, about 0.5 mg to about 25 mg, about 0.5 mg to about 1 mg, about 1 mg to about 10 mg, about 1 mg to about 5 mg, about 3 mg to about 5 mg, or about 4 mg to about 5 mg.

The dosage ranges described herein provide guidance for administering the provided pharmaceutical compositions to adults. The amount administered to, for example, children or adolescents can be determined by one of ordinary skill in the art and may be lower than or the same as that administered to adults.

The compounds or other compositions described herein may be administered in combination with one or more additional agents, or after administration of one or more "second" agents to patients who have developed resistance thereto (as further discussed below). The compounds may be administered in combination with additional agents that increase their bioavailability, reduce and/or alter their metabolism, inhibit their excretion, and/or alter their distribution in the body. The therapies employed may achieve the desired effect for the same condition, and/or they may achieve different effects.

Treatment methods and uses: The compounds and/or other compositions described herein (such as compounds in specified forms and pharmaceutical compositions containing compounds or specified forms thereof, including those compounds and pharmaceutical compositions formulated in unit dosage forms) have Various uses, including in research and/or clinical settings (e.g., in methods of providing diagnosis or prognosis, in selecting patients for treatment, and in preventive or therapeutic methods). For example, in a diagnostic method, a composition of the invention can be applied to a biological sample obtained from a patient (e.g., a sample containing tumor cells), and the contacted sample can be analyzed to determine whether the composition inhibits the activity of CDK7 , thus indicating that the patient suffers from cancer (or other proliferative diseases) caused by excessive activity of CDK7.

In some embodiments, the compounds and other compositions described herein (e.g., pharmaceutical compositions and kits) are used to prevent or treat proliferative diseases (e.g., cancer, benign tumors, or pathological angiogenesis) in patients in need thereof (e.g., patients having cells (e.g., tumor cells) whose proliferation or metastasis can be inhibited by inhibiting CDK7). In various embodiments of these methods, steps of obtaining a biological sample from a patient and/or detecting the presence and/or measuring the amount or activity of CDK7 in a provided biological sample may be performed (i.e., such analysis steps are optional and/or may be performed or may have been performed prior to administering the composition of the invention). Therefore, evaluating a patient as described herein may be performed or may have been performed in a method of selecting a patient for treatment with a composition of the invention. And similarly, in any embodiment of the methods of the invention, diagnostic information can be obtained, for example, by performing or obtaining test results that are suitable for diagnosing whether a patient has a proliferative disease (e.g., cancer) and/or describing the disease (e.g., its type, grade, susceptibility to treatment, etc.) (i.e., diagnostic testing or obtaining diagnostic information is an optional step that can be performed prior to or during treatment of any of the diseases described herein and/or as part of the patient selection process). For example, the methods and uses described herein can be applied to patients who have been determined to have "high-grade" cancer (e.g., high-grade serous ovarian cancer); determined to have tumor cells that exhibit a certain phenotype (e.g., breast cancer cells that are estrogen receptor-positive (ER+) or "triple negative"); and/or determined to have developed resistance to treatment with a previously administered treatment agent (e.g., a chemotherapeutic agent such as another CDK inhibitor (e.g., palbociclib) or a receptor degrader (e.g., fulvestrant)). The methods and uses described herein can include the step of making the determination just mentioned; the step of determining whether the patient has high-grade cancer, tumor cells of a specified phenotype, or has developed resistance to a previously administered treatment agent. The treatment methods entail administering to a patient in need thereof a therapeutically effective amount of a compound described herein (e.g., a compound having a structure depicted in Formula I or a subgenus or species thereof in a pharmaceutically acceptable composition in a form specified herein (e.g., as a salt or mixture of enantiomers) to reduce a sign or symptom of the disease. An analyte (e.g., CDK7 (e.g., its expression or activity), hormone receptor status, or other parameters described above) can be assessed, for example, in a biological sample obtained from a patient at any time point before or during a diagnostic, treatment, or prevention regimen.

Methods of employing the compounds or other compositions described herein and involving the administration of the compounds or compositions to a patient (eg, methods of diagnosis, treatment, or prophylaxis) may also be referred to by the term "use" and vice versa. For example, the present invention encompasses: the use of a compound or composition described herein for the treatment of a disease described herein (e.g., a proliferative disease, such as cancer); for diagnosing a patient, selecting a patient for treatment, and/or treating a disease (e.g., a proliferative disease, such as cancer, or an inflammatory disease, an autoimmune disease, or an autoinflammatory disease); and for use in the treatment of a disease described herein or for use in the preparation of a drug for the treatment of a disease described herein Use of compounds or compositions as agents for diseases such as proliferative diseases such as cancer or autoimmunity.

Methods of the invention involving diagnosis/selection of patients and/or treatment of diseases described herein (e.g., proliferative diseases, such as cancer) may specifically exclude any one or more of the disease types described herein (e.g., any one or more of the cancer types). For example, the invention features a method of treating cancer by administering a compound described herein (e.g., a compound of Formula I or a subgenus or species thereof), with the proviso that the cancer is not breast cancer; with the proviso that the cancer is not breast cancer or leukemia; with the proviso that the cancer is not breast cancer, leukemia, or ovarian cancer, etc., and excluding any one of the disease/cancer types listed herein, and having the same concept of variables as excluding the listed elements associated with other aspects and embodiments of the invention (e.g., chemical substitutions of the compounds or components of the kits and pharmaceutical compositions described herein).

In various embodiments, the individual treated is: a mammal; a human; a domestic or companion animal, such as a dog, cat, cow, pig, horse, sheep, or goat; a zoo animal; or a research animal, such as a rodent, dog, Non-human primates (eg, cynomolgus macaques or rhesus monkeys), or non-human transgenic animals, such as transgenic mice or transgenic pigs. Where the patient is a human, the human may be a male, female, or transgender individual of any age group (eg, a pediatric patient (eg, infant, child, or adolescent) or an adult patient (eg, young adult, middle-aged, or elderly)). Similarly, where the patient is a non-human animal (eg, a mammal), it can be a male or female of any age or stage of development. Birds, especially those of commercial value, are also suitable patients.

The proliferative diseases to be treated or prevented using a compound of formula I or a subgenus or species thereof may be associated with aberrant activity of CDK7. Abnormal activity of CDK7 may be elevated and/or inappropriate (eg, abnormal) activity of CDK7. In certain embodiments, CDK7 is not overexpressed and the activity of CDK7 is elevated and/or inappropriate (e.g., CDK7 has increased activity, additional undesirable activity, resistance to native activity modulation compared to wild-type CDK7) mutated CDK7 that confers resistance to degradation, resistance to degradation, etc.). In certain other embodiments, CDK7 is overexpressed (at the mRNA and/or protein levels), and the activity of CDK7 is elevated and/or inappropriate. Compounds of formula (I) or subgenus or species thereof and pharmaceutically acceptable salts, solvates, stereoisomers, tautomers, isotopic forms and compositions as described herein (i.e., containing A composition of one or more of the foregoing) can inhibit CDK7 activity and is suitable for the treatment and/or prevention of proliferative diseases.

Proliferative diseases may also be associated with the inhibition of apoptosis of cells in a biological sample or individual. Although the present invention is not limited by any potential mechanism of action, it is expected that inhibition of CDK7 activity causes cytotoxicity by inducing apoptosis. The compositions of the present invention can induce apoptosis and are therefore suitable for the treatment and/or prevention of proliferative diseases, particularly proliferative diseases in which CDK7 is overexpressed or overactive.

In certain embodiments, the proliferative disease to be treated or prevented using the compositions of the present invention is cancer. All types of cancers disclosed herein or known in the art are within the scope of the present invention, but in particular those cancers known to be associated with CDK7 activity (e.g., overactivity, overexpression, or misexpression). In various embodiments, the proliferative disease is a cancer associated with: dependence on BCL-2 type anti-apoptotic proteins (e.g., MCL-1 and/or XIAP); overexpression of MYC; overexpression of CDK18; overexpression of CDK19; overexpression of FGFR1; overexpression of CDK6; overexpression of CCND2; or overexpression of CDKN2A. In certain embodiments, the proliferative disease is a cancer associated with the absence or suppression of the wild-type RB1 gene. Thus, and as indicated above, the present invention encompasses methods and uses (eg, methods of treating a patient and/or selecting a patient for treatment) of assessing one or more of the analytes just described.

In certain embodiments, the proliferative disease is a hematological cancer, which may also be referred to as a hematopoietic or blood cancer or malignant disease. More specifically, and in various embodiments, the blood cancer may be a leukemia, such as acute lymphoblastic leukemia (ALL; such as B-cell ALL or T-cell ALL), acute myeloid leukemia (AML; such as B-cell AML or T-cell ALL) AML), chronic myeloid leukemia (CML; such as B-cell CML or T-cell CML), chronic lymphocytic leukemia (CLL; such as B-cell CLL (such as hairy cell leukemia) or T-cell CLL), chronic neutrophilic leukemia Leukemia (CNL) or chronic myelomonocytic leukemia (CMML). Blood cancers can also be lymphomas, such as Hodgkin lymphoma (HL; such as B-cell HL or T-cell HL), non-Hodgkin's lymphoma (NHL; such as B-cell NHL or T-cell NHL) , follicular lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphoma (such as splenic marginal zone B-cell lymphoma), Primary mediastinal B-cell lymphoma (such as splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (also known as Waldenster Waldenstrom's macroglobulinemia), immunoblastic large cell lymphoma, precursor B lymphoblastic lymphoma, or primary central nervous system (CNS) lymphoma. B-cell NHL can be diffuse large cell lymphoma (DLCL; eg, diffuse large B-cell lymphoma), and T-cell NHL can be precursor T lymphoblastic lymphoma or peripheral T-cell lymphoma (PTCL). In turn, PTCL can be cutaneous T-cell lymphoma (CTCL), such as mycosis fungoides or Sezary syndrome, angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathies Type T cell lymphoma, subcutaneous anniculitis-like T cell lymphoma, or pleomorphic large cell lymphoma. Although the present invention is not limited to the treatment or prevention of hematological cancers of any particular cause or manifestation, stem cells within the bone marrow can proliferate, thereby becoming the predominant cell type within the bone marrow and targeted by the compounds described herein. Leukemia cells can accumulate in the blood and infiltrate organs such as lymph nodes, spleen, liver and kidneys. In some embodiments, compounds of the invention are useful in the treatment or prevention of leukemia or lymphoma.

In other embodiments, the proliferative disease is characterized by a solid tumor, either as a primary site or a metastatic site. For example, in various embodiments, the cancer or tumor diagnosed, treated, or prevented as described herein is an acoustic neuroma; a malignant adenoma; an adrenal cancer; anal cancer; angiosarcoma (e.g., lymphangiosarcoma, lymphoendothelial sarcoma, angiosarcoma); coccyx cancer; benign monoclonal globulinemia (also known as monoclonal globulinemia of undetermined significance (MGUS); bile duct cancer (e.g., cholangiocarcinoma); bladder cancer; breast cancer (e.g., breast cancer, papillary carcinoma, breast cancer, medullary breast cancer; any of which may be present in an individual with a specific type, such as HR+ (ER+ or PR+), HR- (no estrogen and progesterone receptors), triple negative breast cancer (TNBC; ER-/PR-/HER2-) or triple positive breast cancer); Brain cancer (e.g., meningioma, neuroglioma, neuroglioma (e.g., astrocytoma, oligodendroglioma, medulloblastoma); bronchial cancer; carcinoid tumors, which may be benign; cervical cancer (e.g., cervical gland carcinoma); choriocarcinoma; chordoma; craniopharyngioma; colorectal cancer (e.g., colon cancer, rectal cancer, or colorectal adenocarcinoma); connective tissue cancer; epithelial cancer; ependymoma; endothelial sarcoma (e.g., Kaposi's sarcoma or multiple idiopathic hemorrhagic sarcomas); endometrial cancer (e.g., uterine cancer, uterine sarcoma); esophageal cancer (e.g., esophageal adenocarcinoma, Barrett's adenocarcinoma); Ewing's sarcoma (or other pediatric sarcomas such as embryonal rhabdomyosarcoma or alveolar rhabdomyosarcoma); eye cancer (e.g., intraocular melanoma, retinoblastoma); common hypereosinophilic leukemia; gallbladder cancer; stomach cancer (e.g., gastric adenocarcinoma); gastrointestinal stromal tumors (GIST); germ cell cancer; head and neck cancer (e.g., head and neck squamous cell carcinoma, oral cancer (e.g., oral squamous cell carcinoma), pharyngeal cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer)); hypopharyngeal cancer; inflammatory myofibroblastoma; immunocytic amyloid degeneration; kidney cancer (e.g., nephroblastoma, also known as Wilms' tumor) tumor), kidney cell cancer); liver cancer (e.g., hepatocellular carcinoma (HCC), malignant hepatocellular carcinoma); lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), lung adenocarcinoma); leiomyosarcoma (LMS); mastocytosis (e.g., systemic mastocytosis); muscle cancer; myelodysplastic syndrome (MDS); mesothelioma; myeloproliferative disorders (MPDs) (e.g., polycythemia vera (PV), essential thrombocythemia (ET), myelofibrosis of unknown cause (AMM) also known as myelofibrosis (MF), chronic idiopathic myelofibrosis, hypereosinophilic syndrome (HES)); neuroblastoma; neurofibroma (e.g., neurofibroma (NF) type 1 or 2, schwannomatosis); neuroendocrine cancer (e.g., gastrointestinal pancreatic neuroendocrine tumor (GEP-NET), carcinoid tumor); osteosarcoma (e.g., bone cancer); ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma; HGSOC; epithelial ovarian cancer (e.g., ovarian clear cell carcinoma); papillary adenocarcinoma; pancreatic cancer (e.g., pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), islet cell tumor); penile cancer (e.g., Paget's disease of the penis and scrotum); pineal tumor; primary neuroectodermal tumor (PNT); plasma cell tumor; preneoplastic syndrome; prostate cancer (e.g., prostate adenocarcinoma); rhabdomyosarcoma; salivary gland cancer; skin cancer (eg, squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)); small bowel cancer/small intestine cancer); soft tissue sarcoma (e.g., malignant fibrohistomas (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma); sebaceous gland carcinoma; sweat gland carcinoma; synovioma; testicular cancer (e.g., seminoma, testicular embryonal carcinoma); thyroid cancer (e.g., papillary thyroid carcinoma, papillary thyroid carcinoma (PTC), medullary thyroid carcinoma); urethral cancer; vaginal cancer; and vulvar cancer (e.g., Paget's disease of the vulva). In some embodiments, the proliferative disease is associated with pathological angiogenesis, and the methods of the invention encompass inhibiting pathological angiogenesis in the context of cancer treatment.

In some embodiments, the proliferative disease is an inflammatory disease. It is contemplated that all types of inflammatory diseases disclosed herein or known in the art are within the scope of the present invention. In some embodiments, the inflammatory disease is rheumatoid arthritis. In some embodiments, the proliferative disease is an autoinflammatory disease. It is contemplated that all types of autoinflammatory diseases disclosed herein or known in the art are within the scope of the present invention. In some embodiments, the proliferative disease is an autoimmune disease. It is contemplated that all types of autoimmune diseases disclosed herein or known in the art are within the scope of the present invention.

Biological cells can be evaluated (e.g., in the context of diagnostic and/or patient selection methods) or treated in vitro or in vivo. In various embodiments, the cells are: proliferating cells; blood cells; lymphocytes; cancer cells; leukemia cells; CLL cells; melanoma cells; multiple myeloma cells; benign proliferative cells; endothelial cells; and/or immune cells.

In another aspect, the present invention provides a method of down-regulating CDK7 expression in a biological sample or subject by administering to the sample or subject an effective amount of a compound described herein or a salt, solvate, stereoisomer, tautomer or isotopic variant thereof. The compound or a specified form thereof (e.g., a salt of the compound or a stereoisomer or isotopic variant of the compound or its salt) can be in a pharmaceutical composition (e.g., as further described herein).

Treatment or prevention methods and "uses" described herein may include combining one or more additional therapeutically active agents (i.e., a "second" agent that is different from a compound or other composition of the invention) with a composition of the invention (e.g., , a compound of formula I, its pharmaceutically acceptable salts, solvates, stereoisomers, tautomers or isotopic forms, or containing such compounds or their designated forms (e.g., their pharmaceutically acceptable Salt) composition in combination with the step of administering. We may refer to such methods and uses as "combination therapy." The second agent includes, but is not limited to, antiproliferative agents, anticancer agents, antidiabetic agents, anti-inflammatory agents, immune Inhibitors and analgesics. The second agent may, but does not necessarily, synergistically enhance the inhibition of CDK7 induced by a compound or composition of the invention (i.e., the "first" agent) in a biological sample or individual. Accordingly, the first and second agents The combination of two agents may be suitable for use of the second agent in the absence of the first agent to treat a proliferative disease that is resistant to treatment. In this case, and as discussed above, the patient may be administered after it has been established that he/she is resistant to the previous administration. The composition of the present invention is administered after resistance to the therapeutic agent has developed. CDK7 overexpression is associated with hormone receptor-positive breast cancer (HR ⁺ breast cancer), triple-negative breast cancer (TNBC), acute myeloid leukemia (AML), and small cell lung cancer (SCLC). ), esophageal squamous cell carcinoma, neuroblastoma, high-grade glioma, ovarian cancer, solid tumors and other hematological malignancies.

In the combination therapy of the present invention, the compound of formula (I), (Ia) or species thereof or pharmaceutically acceptable salts, solvates, stereoisomers, tautomers or isotopic forms thereof may be administered simultaneously with, before or after one or more additional (i.e., different) therapeutic agents. Each additional therapeutic agent may be administered in a dose and/or according to a dosing regimen determined for that particular agent. The additional therapeutic agents may also be administered together with each other and/or with the compounds or compositions described herein in a single dose, or separately in different doses. The specific combination employed in the regimen will take into account the compatibility of the compounds of the present invention with the additional therapeutic agents and/or the desired therapeutic and/or preventive effect to be achieved. Generally speaking, it is contemplated that the additional therapeutic agent used in combination is used in an amount not exceeding the amount used alone. In some embodiments, the amount used in the combination will be lower than the amount used alone.

The second agent may be, but is not limited to, an antiproliferative agent, an anticancer agent, an antidiabetic agent, an anti-inflammatory agent, an immunosuppressant, or an analgesic agent. Such therapeutic agents include small organic molecules such as drug compounds (e.g., compounds approved by the U.S. Food and Drug Administration or the European Medicines Agency), polypeptides (including nucleoproteins, mucins, glycoproteins, lipoproteins, and antibodies, and whether the polypeptide is synthetic or naturally occurring), carbohydrates (e.g., mono-, oligo-, and polysaccharides), small molecules (linked to proteins, steroids, nucleotides, nucleosides, and nucleic acids (e.g., DNA and RNA, including any RNA for RNAi configurations, regardless of length)).

In certain embodiments, the additional therapeutic agent is a Bcl-2 inhibitor, such as APG-1252, APG-2575, BP1002, S55746, or venetoclax; a CDK9 inhibitor, such as alvocidib, AT7519, AZD5576, BAY1251152, BAY1143572, CYC065, nanoflavopiridol, NVP2, seliciclib (CYC202), TG02, TP-1287, VS2-370, or voruciclib (formerly P1446A-05). Hormone receptor (e.g., estrogen receptor) degraders, such as Faslodex® (fulvestrant); Flt3 (FMS-like tyrosine kinase 3) inhibitors, such as CDX-301, CG'806, CT053PTSA; crenolanib besylate, ENMD-2076, FF-10101-01, FLYSYN, gilteritinib (ASP2215), HM43239, Iclusig® (ponatinib), NMS-088, Nexavar® (sorafenib), pacritinib, pexidartinib/PLX3397, quizartinib, Rydapt® (midostaurin), SEL24, SKI-G-801, or SKLB1028; PARP inhibitors, such as Lynparza® (olaparib), Rubraca® (rucaparib), Talzenna® (talazoparib), veliparib (ABT-888), or Zejula® (niraparib); BET inhibitors, such as ABBV-075, BAY-299, BAY-1238097, BMS-986158, CPI-0610, CPI-203, FT-1101, GS-5829, GSK-2820151, GSK-525762, I-BET151, I-BET762, INCB054 329, JQ1, MS436, OTX015, PFI-1, PLX51107, RVX2135, TEN-010, ZEN-3694, or compounds disclosed in U.S. Application No. 12/810,564 (now U.S. Patent No. 8,476,260), which is incorporated herein by reference in its entirety; platinum-based therapeutic agents, such as cis-platinum, Eloxatin® (oxaliplatin), nedaplatin, Paraplatin® (carboplatin), phenanthriplatin, picoplatin, satraplatin (JM216), or triplatinum tetranitrate; or a CDK4/6 inhibitor such as BPI-1178, G1T38, Ibrance® (palbohib), Kisqali® (ribociclib), ON 123300, trilaciclib, or Verzenio® (abemaciclib).

In one embodiment, a method of treatment includes administering to a patient suffering from a sarcoma (e.g., osteosarcoma, rhabdomyosarcoma, or Ewing's sarcoma) a therapeutically effective amount of a compound described herein (e.g., a compound of Formula I (e.g., Compounds 100, 101 or 102)) and therapeutically effective amounts of PARP inhibitors (e.g., Lynparza® (olaparib), Rubraca® (rucapanib), Talzenna® (razopanib), veliparib (ABT-888) ) or Zejula® (niraparib).

Kits (eg medical kits) are also encompassed by the present invention. The kit may be used to prevent and/or treat any of the diseases described herein. Kits provided may include a pharmaceutical composition or compound of the invention; and a container (such as a vial, ampoule, bottle, syringe and/or dispenser package or other) for storage, reconstitution and/or administration of the compound or composition. suitable for the container). In some embodiments, the provided kit optionally further includes a second container containing a pharmaceutical excipient for diluting or suspending the pharmaceutical composition or compound of the invention. In some embodiments, the pharmaceutical composition or compound provided in the container and the second container are combined to form a unit dosage form. In some embodiments, the provided kit optionally further includes a second or third container containing additional therapeutic agents to be administered in combination with the pharmaceutical compositions or compounds of the invention.

In some embodiments, the kit is suitable for preventing and/or treating a proliferative disease in an individual. In some embodiments, the kit further comprises instructions for administering the compound or its pharmaceutically acceptable salt, solvate, hydrate, tautomer, isotope and labeled derivative or pharmaceutical composition thereof to an individual for preventing and/or treating a proliferative disease.

In another embodiment, the present invention provides compounds of Formula I or Formula Ia and pharmaceutically acceptable salts, solvates and hydrates thereof for treating a proliferative disease in an individual. In certain embodiments, the present invention provides compounds described herein and pharmaceutically acceptable salts and compositions thereof for treating a proliferative disease in an individual. In certain embodiments, the present invention provides compounds described herein and pharmaceutically acceptable salts and compositions thereof for inhibiting cell growth. In certain embodiments, the present invention provides compounds described herein and pharmaceutically acceptable salts and compositions thereof for inducing apoptosis in a cell. In certain embodiments, the present invention provides compounds described herein and pharmaceutically acceptable salts and compositions thereof for inhibiting transcription. Examples

The compounds described herein can be prepared from readily available starting materials and according to the synthetic schemes described below. Alternatively, one of ordinary skill in the art can readily modify the disclosed schemes. For example, it should be understood that where process conditions (e.g., reaction temperature, reaction time, molar ratio of reactants, solvents, pressure, etc.) are given, other process conditions may also be used.

In addition, and as will be apparent to one of ordinary skill in the art, protecting groups can be used to prevent certain functional groups from undergoing undesired reactions. The selection of suitable protecting groups for particular functional groups and suitable conditions for protection and deprotection is well known in the art. For example, Greene et al. disclose many protecting groups and guidance for their introduction and removal ( Protecting Groups in Organic Synthesis , 2nd edition, Wiley, New York, 1991, and references cited therein).

Example 1 : Synthesis of (2R,5R)-5- amino - 2- methyl - piperidine -1- carboxylic acid benzyl ester and (2S,5S)-5- amino -2- methyl - piperidine- Benzyl 1- carboxylate Step 1 : Benzyl 5-( tert-butoxycarbonylamino )-2- methyl - piperidine -1 -carboxylate

We added a solution containing commercially available racemic trans-tert-butyl N-(6-methyl-3-piperidinyl)carbamate (5 g, 23.33 mmol, 1 equiv) and NaHCO ₃ ( To a solution of 13.72 g, 163.32 mmol, 7 equiv) in tetrahydrofuran (THF; 50 mL) and H ₂ O (50 mL) was added dropwise CbzCl (5.97 g, 35.00 mmol, 4.98 mL, 1.5 equiv). The mixture was stirred at 15°C for 2 hours, then poured into water (50 mL) and extracted with ethyl acetate (EtOAc; 50 mL×3). The combined organic layers were washed with brine (50 mL× ₃ ), dried over _Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by medium pressure liquid chromatography (MPLC; SiO ₂ , PE:EtOAc=5:1 to 1:1) to obtain the title compound as a yellow solid (9.7 g, 18.04 mmol, 77.32% yield, 64.8% purity). Step 2 : (2R,5R)-5- amino -2- methyl - piperidine -1- carboxylic acid benzyl ester and (2S,5S)-5- amino -2- methyl - piperidine -1 -Benzyl formate :

We racemicized trans benzyl 5-(tert-butoxycarbonylamino)-2-methyl-piperidine-1-carboxylate (9.7 g, 27.84 mmol, 1 equiv) in EtOAc (100 mL) To the mixture was added HCl/EtOAc (15 mL, 4 M), and the mixture was stirred at 15 °C for 1 h. We then filtered the mixture and collected the filter cake. The solid was dissolved in methanol (MeOH; 15 mL) and the pH was adjusted to 9 using AMBERLYST® A21 before the mixture was filtered and the filtrate was concentrated. The residue was subjected to supercritical fluid chromatography (SFC; column: sold by Daicel as CHIRALCEL® (chemicals used in chromatography) ODH (250 mm × 30 mm, 5 µm); mobile phase: [0.1% NH _3. H ₂ O MeOH]; B%: 28%-28%, 16 min) was separated to obtain the title compound 1 (1.9 g, SFC: Rt=2.264 min, 93.2% ee, peak 1) as a light yellow solid. ) and the title compound 2 (1.9 g, SFC: Rt=2.593 min, 98.6% ee, peak 2). Peak 1 is structure 3 . Peak 2 is structure 4 .

Example 2 : Synthesis of 7- dimethylphosphonyl -3-[2-[[(3S,6S)-6- methyl - 3- piperidinyl ] amino ]-5-( trifluoromethyl ) pyrimidine -4- yl ]-1H- indole -6- carbonitrile ( Compound 100) Step 1 : (2S,5S)-5-[[4-(7- chloro -6- cyano -1H- indole -3 -yl )-5-( trifluoromethyl ) pyrimidin -2 -yl ] amino ]-2- methyl - piperidine - 1- carboxylic acid benzyl ester

A mixture of 7-chloro-3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-1H-indole-6-carbonitrile (0.81 g, 2.27 mmol, 1 eq.), (2S,5S)-5-amino-2-methyl-piperidine-1-carboxylic acid benzyl ester (732.20 mg, 2.95 mmol, 1.3 eq.) and N,N-diisopropylethylamine (DIEA or DIPEA; 879.41 mg, 6.80 mmol, 1.19 mL, 3 eq.) in N-methyl-2-pyrrolidone (NMP; 8 mL) was stirred at 140° C. for 1 hour. The reaction mixture was diluted with H ₂ O (100 mL) and extracted with EtOAc (50 mL×2). The combined organic layers were washed with brine (100 mL x 2), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 10:1 to 4:1) to give the title compound (1.1 g) as a yellow solid.

Step 2 : (2S,5S)-5-[[4-(6- cyano -7 -dimethylphosphatidyl -1H- indol -3- yl )-5-( trifluoromethyl ) pyrimidin -2- yl ] amino ]-2- methyl - piperidine -1-carboxylic acid benzyl ester

We prepared (2S,5S)-5-[[4-(7-chloro-6-cyano-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidine- 2-yl]Amino]-2-methyl-piperidine-1-carboxylic acid benzyl ester (1.05 g, 1.85 mmol, 1 equivalent), methylphosphonylmethane (720.17 mg, 9.23 mmol, 5 equivalents), K ₃ PO ₄ (783.45 mg, 3.69 mmol, 2 equiv), Pd(OAc) ₂ (41.43 mg, 184.54 µmol, 0.1 equiv), Xantphos (C ₃₉ H ₃₂ OP ₂ ; 106.78 mg, 184.54 µmol, 0.1 equiv) and A mixture of dimethylformamide (DMF; 10 mL) was degassed and flushed with N ₂ (×3). The mixture was then stirred in the microwave at 150°C for 1 hour. The reaction mixture was diluted with _H2O (100 mL) and extracted with ethyl acetate (EtOAc; 50 mL×3). The combined organic layers were washed with brine (150 mL×2), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain a residue, which was analyzed by column chromatography (SiO ₂ , petroleum ether/ethyl acetate). Ester = 10:1 to 1:1) to obtain the title compound (490 mg) as a yellow oil.

Step 3 : 7- Dimethylphosphonyl -3-[2-[[(3S,6S)-6- methyl - 3- piperidinyl ] amino ]-5-( trifluoromethyl ) pyrimidine- 4- yl ]-1H- indole -6- carbonitrile

(2S,5S)-5-[[4-(6-cyano-7-dimethylphosphono-1H-indol-3-yl)-5-(trifluoromethyl) under _N To a solution of pyrimidin-2-yl]amino]-2-methyl-piperidine-1-carboxylic acid benzyl ester (440 mg, 720.64 µmol, 1 equiv) in EtOAc (5 mL) was added Pd/C (200 mg, 10% purity). We degassed the suspension under vacuum, purged it several times with _H2 , then stirred the mixture under _H2 (15 psi) at 20°C for 3 hours before filtering it. The filtrate was concentrated to obtain a residue, which was purified by prep-HPLC (high performance liquid chromatography; neutral conditions) to obtain the title compound (142.2 mg) as a white solid.

The reaction was combined with another reaction at 50 mg scale for purification by liquid chromatography mass spectrometry (LCMS). LCMS: ET6034-1492-P1C: (M+H ⁺ ): 477.1 @2.572 (10-80% ACN (acetonitrile) in H ₂ O, 4.5 min). ¹ H NMR (400 MHz, DMSO (dimethylsulfoxide)-d6) δ 8.74 (br d, J = 7.89 Hz, 1H), 8.65-8.44 (m, 2H), 8.17 (br d, J = 15.35 Hz, 1H), 7.84 (br t, J = 8.11 Hz, 1H), 7.67 (br t, J = 7.02 Hz, 1H), 3.81 (br s, 1H), 3.10 (br d, J =11.40 Hz, 1H), 2.45-2.38 (m, 1H), 2.02 (d, J = 13.59 Hz, 8H), 1.64 (br d, J = 11.40 Hz, 1H), 1.49-1.34 (m, 1H), 1.11 (br d, J = 10.96 Hz, 1H), 0.97 (br d, J = 5.70 Hz, 3H).

Example 3 : Synthesis of (S)-6,6 -dimethylpiperidin - 3- amine

We dissolved ( S )-tert-butyl (6-oxopiperidin-3-yl)carbamate (1.00 g, 4.67 mmol) ( Tetrahedron Letters , 36 :8205, 1995) in THF (47 mL) and cooled the solution to -10°C. Zirconium (IV) chloride (2.61 g, 11.22 mmol) was added, and the mixture was stirred at this temperature for 30 minutes. Methylmagnesium bromide solution (3 M in diethyl ether, 20.25 mL, 60.75 mmol) was added, and the reaction mixture was slowly warmed to room temperature and stirred at room temperature overnight. The solution was quenched with 30% aqueous NaOH, diluted with EtOAc, filtered and then extracted 3 times with EtOAc. The organic phases were combined, dried over sodium sulfate, filtered and concentrated in vacuo to give the crude product as a yellow oil, which was used without purification. The oil was dissolved in dichloromethane (DCM; 47 mL) and trifluoroacetic acid (TFA; 3.58 mL, 46.73 mmol) was added. The reaction mixture was stirred at room temperature for 16 hours, concentrated in vacuo and co-evaporated with DCM several times to give the crude title compound as a brown oil, which was used in the next step without further purification.

Example 4 : Synthesis of (S)-7-( dimethylphosphinoyl )-3-(2-((6,6 -dimethylpiperidin -3 -yl ) amino )-5-( trifluoromethyl ) pyrimidin -4- yl )-1H- indole -6 - carbonitrile ( Compound 101) . Step 1 : 7- Bromo -1H- indole -6- carboxylic acid

We stirred a solution of magnesium vinyl bromide (1.0 M in THF (159 mL, 159 mmol) at -78 °C and added a solution of 2-bromo-3-nitrobenzoic acid (10.0 g, 39.8 mmol) in THF (159 mL) dropwise over a period of 1 hour. The reaction mixture was allowed to reach room temperature and stirred at that temperature overnight. The reaction mixture was then poured onto saturated aqueous ammonium chloride solution (150 mL) and acidified to pH 2 using 1 M aqueous HCl. The crude product was extracted with EtOAc (3 x 200 mL), the extract was dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was then washed with DCM (100 mL). The product was wet-triturated with 10% ethyl acetate (20 mL) and dried overnight with air flow to obtain the title compound (7.58 g, 31.58 mmol, 79% yield) as a light brown solid. Step 2 : 7- bromo -1H- indole -6- carboxamide

We stirred a solution of 7-bromo-1H-indole-6-carboxylic acid (6.58 g, 27.4 mmol) in DMF (54.8 mL) at 0°C, and added 1,1'-carbonyldiimidazole (CDI; 8.89 g, 54.8 mmol) portionwise thereto. The mixture was stirred for 5 minutes, and an intermediate was observed by LCMS. We then added NH4OH (39.5 mL, 274 mmol) at 0°C, and stirred the solution for 5 minutes. The reaction was quenched with saturated aqueous ammonium chloride (25 mL) and saturated aqueous sodium chloride (25 mL), and then diluted with 2-methyltetrahydrofuran (MeTHF; 50 mL). We separated the phases and washed the organic layer again with saturated aqueous ammonium chloride (25 mL) and saturated aqueous sodium chloride (25 mL). The organic layer was then dried over sodium sulfate, filtered and concentrated in vacuo to give the title compound, which was carried forward to the next step in assuming quantitative yield. Step 3 : 7- Bromo -1H- indole -6- carbonitrile

To a suspension of 7-bromo-1H-indole-6-carboxamide (7.53 g, 31.5 mmol) in DCM (315 mL) at 0°C, we added _Et3N (triethylamine; 44.1 mL, 315 mmol), and stirred the resulting orange solution at that temperature until we obtained a homogeneous solution. MsCl (12.2 mL, 157 mmol) was then added dropwise, and the solution was stirred at 0°C for 5 minutes. We diluted the mixture with ethyl acetate and washed with saturated aqueous sodium bicarbonate solution, then extracted the aqueous layer two more times with ethyl acetate. The organic layers were combined, washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by filtering it through a pad of silica (eluting with ethyl acetate) to give the title compound as a brown solid (5.80 g, 26.24 mmol, 83% yield). Step 4 : 7- Bromo -3-(2- chloro -5-( trifluoromethyl ) pyrimidin -4- yl )-1H- indole -6- carbonitrile

We added AlCl ₃ (1.83 g, 13.6 mmol) to a solution of 2,4-dichloro-5-trifluoromethylpyrimidine (3.66 mL, 27.2 mmol) in 1,2-dichloroethane (DCE; 36.2 mL) and stirred the resulting suspension at 80° C. for 30 minutes. We added 7-bromo-1H-indole-6-carbonitrile (2.00 g, 9.05 mmol) to the mixture and stirred the resulting red solution at 80° C. until complete conversion (4 hours). The reaction mixture was then diluted with MeTHF (100 mL) and washed with water (100 mL). The aqueous layer was extracted with 2-MeTHF (100 mL), and the organic extracts were combined, dried over sodium sulfate, filtered and concentrated in vacuo. The formation of two possible regioisomers was observed in a ratio of 3:1 (desired/undesired). The residue was purified by reverse phase chromatography on C18 (MeCN(acetonitrile)/water, 15-80% gradient) to afford the title compound (1.51 g, 3.76 mmol, 42% yield) as a beige solid. ¹ H NMR (500 MHz, DMSO) δ 13.00 (brs, 1H), 9.17 (s, 1H), 8.35 (d, J = 8.4 Hz, 1H), 8.16 (d, J = 2.6 Hz, 1H), 7.71 (d, J = 8.4 Hz, 1H). Step 5 : (S)-7- Bromo -3-(2-((6,6 -dimethylpiperidin -3- yl ) amino )-5-( trifluoromethyl ) pyrimidin -4- yl )-1H- indole -6- carbonitrile

We combined 7-bromo-3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carbonitrile (200 mg, 0.498 mmol), ( S )- 6,6-Dimethylpiperidin-3-amine (95.8 mg, 0.747 mmol) and DIPEA (174 µL, 0.996 mmol) were dissolved in NMP (4 mL), and the reaction mixture was stirred in an oil bath at 130 °C. until complete conversion (3 hours). The mixture was cooled to room temperature, loaded directly onto a C18 column, and purified by reverse phase chromatography (MeCN with 0.1% FA (formic acid) in water also containing 0.1% FA, 0 to 100% gradient). The title compound was obtained as a beige solid (245 mg, 0.497 mmol, quantitative yield). Step 6 : (S)-7-( dimethylphosphonyl )-3-(2-((6,6 -dimethylpiperidin -3 -yl ) amino )-5-( trifluoromethyl ) -pyrimidin -4- yl )-1H- indole -6- carbonitrile

We prepared ( S )-7-bromo-3-(2-((6,6-dimethylpiperidin-3-yl)amino)-5-(trifluoro) in a 2.5 mL microwave vial under nitrogen. Methyl)-pyrimidin-4-yl)-1H-indole-6-carbonitrile (180.0 mg, 0.365 mmol), Xantphos (21.5 mg, 36.5 µmol), palladium(II) acetate (4.14 mg, 18.2 µmol) and K ₃ PO ₄ (85.2 mg, 0.401 mmol) combination. Dimethylphosphine oxide (73 mg, 0.912 mmol) was dissolved in anhydrous DMF (1 mL) and the solution was degassed before being combined with the other reactants in a microwave vial. The sealed vial with the reaction mixture was then heated in a microwave reactor at 150°C for 45 minutes. The reaction mixture was cooled to room temperature, loaded directly onto a C18 column, and purified by reverse phase chromatography (MeCN/10 mM aqueous ammonium formate, pH 3.8, 15 to 35% gradient). The title compound was obtained as an off-white solid (76 mg, 0.155 mmol, 42% yield).

Example 5 : Synthesis of (3S)-1- phenylmethyl -5,5- dimethyl - piperidin -3- amine Step 1 : (2S)-5- Pendantoxypyrrolidine -2- carboxylic acid methyl ester

_SOCl2 (215.62 g, 1.81 mol, 131.47 mL, 2 eq) was added to a solution of (2S)-5-oxopyrrolidine-2-carboxylic acid (117 g, 906.18 mmol, 1 eq) in MeOH (500 mL) at 0 °C. The mixture was stirred at 18 °C for 1 hour, then the reaction mixture was concentrated. The residue was diluted with EtOAc (1000 mL) and TEA (triethylamine; 150 mL) and the solid formed was filtered. The filtrate was evaporated to give the title compound (147 g, crude) as a light yellow oil, which was used directly in the next step without any further purification. Step 2 : (S)-5- oxopyrrolidine -1,2- dicarboxylic acid 1- tert-butyl ester 2- methyl ester

To a solution of methyl (2S)-5-oxopyrrolidine-2-carboxylate (147 g, 1.03 mol, 1 eq.), DMAP (4-dimethylaminopyridine; 15.06 g, 123.24 mmol, 0.12 eq.) and TEA (259.80 g, 2.57 mol, 357.35 mL, 2.5 eq.) in EtOAc (500 mL) was added tert-butyl tert-butoxycarbonyl carbonate (291.37 g, 1.34 mol, 306.71 mL, 1.3 eq.) dropwise at 0° C. The mixture was then stirred at 20° C. for 16 hours. The reaction mixture was then washed with HCl (0.5 M, 1000 mL), saturated NaHCO ₃ (1000 mL), brine (1500 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, which was then purified by recrystallization from methyl tert-butyl ether (MTBE; 250 mL). The reaction mixture was filtered and evaporated to give the title compound as a white solid (obtained in 2 batches; batch 1: 108 g, 100% HPLC purity; batch 2: 53 g, 90% ¹ H NMR purity). Step 3 : (S)-4,4 -dimethyl -5- oxopyrrolidine -1,2- dicarboxylic acid 1- tert-butyl ester 2- methyl ester

To (S)-5-Pendant oxypyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (20 g, 82.22 mmol, 1 equiv) at -78 °C under _N2 atmosphere To a solution in THF (500 mL) was added LiHMDS (lithium bis(trimethylsilane)amide; 1 M, 172.66 mL, 2.1 equiv) dropwise. After the addition, the mixture was stirred at this temperature for 30 minutes, followed by the dropwise addition of _CH3I (35.01 g, 246.65 mmol, 15.36 mL, 3 equiv) at -78°C under _N2 atmosphere. The resulting mixture was stirred at 20°C for 2.5 hours. The reaction mixture was diluted with saturated aqueous NH ₄ Cl solution (1000 mL) and extracted with EtOAc (300 mL×3). The combined organic layers were washed with brine (500 mL), _dried over _Na2SO4 , filtered and concentrated under reduced pressure to give a residue, which was analyzed by MPLC ( _SiO2 , PE:EtOAc=4:1-3:1 ) was purified to obtain the title compound (8 g, 25.95 mmol, 31.56% yield, 88% purity) as a light yellow solid. Step 4 : N-[(1S)-4- Hydroxy -1-( hydroxymethyl )-3,3- dimethyl - butyl ] carbamic acid tert-butyl ester

To (S)-4,4-dimethyl-5-pendantoxypyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (4.3 g, _15.85 To a solution of NaBH ₄ (1.80 g, 47.55 mmol, 3 equiv) in THF (35 mL) was added portionwise. After the addition, EtOH (ethanol; 8.25 g, 179.09 mmol, 10.47 mL, 11.3 equiv) was added dropwise at 0°C. The resulting mixture was stirred at 20°C for 16 hours, then poured into saturated aqueous NH ₄ Cl solution (250 mL) and extracted with EtOAc (100 mL×3). The combined organic layers were washed with brine (250 mL), dried over _{Na2SO4 ,} filtered and concentrated under reduced pressure to give the title compound as a colorless oil (3.67 g, crude) which was obtained without any further purification. Use it directly in the next step. Step 5 : Methanesulfonate [(2S)-2-( tert-butoxycarbonylamino )-4,4- dimethyl -5 - methylsulfonyloxy - pentyl ] ester

To tert-butyl N-[(1S)-4-hydroxy-1-(hydroxymethyl)-3,3-dimethyl-butyl]carbamate (3.67 g, 14.84 mmol, 1 To a solution of methanesulfonyl chloride (5.10 g, 44.52 mmol, 3.45 mL, 3 equiv) in EtOAc (25 mL) and TEA (6.01 g, 59.35 mmol, 8.26 mL, 4 equiv) in EtOAc (25 mL) was added dropwise. The resulting mixture was stirred at 20°C for 12 hours and then poured into _H2O (200 mL). The product was extracted using EtOAc (50 mL×3). The organic layer was washed with brine (30 mL), dried _{over Na2SO4} , filtered and evaporated to give the title compound as a colorless oil (6.06 g crude product) which was used directly in the next step without any further purification. . Step 6 : N-[(3S)-1- benzyl -5,5- dimethyl -3- piperidyl ] carbamic acid tert-butyl ester

A flask was charged with methanesulfonic acid [(2S)-2-(tert-butoxycarbonylamino)-4,4-dimethyl-5-methyl-sulfonyloxypentyl] ester (6.06 g, 15.02 mmol, 1 eq.), phenylmethylamine (5.15 g, 48.06 mmol, 5.24 mL, 3.2 eq.) and dimethoxyethane (DME; 50 mL). The reaction mixture was heated to 70 °C for 16 h and then poured into H ₂ O (40 mL). The product was extracted with DCM (40 mL×3). The organic layer was washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered and evaporated to give a crude product, which was purified twice by MPLC (SiO ₂ , PE:EtOAc=20:1-10:1) to give the title compound (580 mg, 1.49 mmol, 9.91% yield, 81.7% purity) as a colorless oil. Step 7 : (3S)-1- Benzyl -5,5- dimethyl - piperidin -3- amine

The flask was charged with tert-butyl N-[(3S)-1-benzyl-5,5-dimethyl-3-piperidinyl]carbamate (300 mg, 942.05 µmol, 1 eq) in HCl/EtOAc (15 mL). The mixture was stirred at 25 °C for 1 h, after which some white precipitate formed. The mixture was filtered and the filter cake was washed with EtOAc (5 mL), collected and dried under vacuum to give the title compound as a white solid (220 mg, 738.23 µmol, 78.36% yield, 85.5% purity, HCl) which was used directly in the next step.

Example 6 : Synthesis of (S)-7-( dimethylphosphonyl )-3-(2-((5,5 -dimethylpiperidin -3 -yl ) amino )-5-( trifluoromethyl ) pyrimidin -4- yl )-1H- indole -6- carbonitrile ( Compound 102) Step 1 : (S)-3-(2-((1- benzyl -5,5- dimethylpiperidin -3- yl ) amino )-5-( trifluoromethyl ) pyrimidin -4- yl )-7 - bromo -1H- indole -6- carbonitrile We dissolved 7-bromo-3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carbonitrile (168 mg, 0.418 mmol), ( S )-1-benzyl-5,5-dimethylpiperidin-3-amine (128 mg, 0.585 mmol) and DIPEA (221 µL, 1.26 mmol) in NMP (2 mL). We stirred the reaction mixture in an oil bath at 130 °C until complete conversion (4 h). The mixture was cooled to room temperature, diluted with EtOAc and washed with saturated aqueous LiCl solution. The organic layer was separated, dried over sodium sulfate, filtered and concentrated in vacuo to give the crude title compound (240 mg, 0.411 mmol, quantitative yield) which was used in the next step without further purification. Step 2 : (S)-3-(2-((1- benzyl -5,5- dimethylpiperidin -3 -yl ) amino )-5-( trifluoromethyl ) pyrimidin -4- yl )-7-( dimethylphosphatyl )-1H- indole -6- carbonitrile

We combined ( S )-3-(2-((1-benzyl-5,5-dimethylpiperidin-3-yl)amino)-5-(trifluoro-methyl)pyrimidin-4-yl)-7-bromo-1H-indole-6-carbonitrile (240 mg, 0.411 mmol), Xantphos (24.3 mg, 41.1 µmol), Palladium(II) acetate (4.66 mg, 20.6 µmol) and _K3PO4 (96.0 mg, 0.452 mmol) in a 2.5 mL microwave vial under nitrogen. _{Dimethylphosphine} oxide (39.2 mg, 0.494 mmol) was dissolved in anhydrous DMF (1 mL) and the solution was degassed before combining with the other reactants in a microwave vial. The sealed vial with the reaction mixture was then heated in a microwave reactor at 145 °C for 45 minutes. The reaction mixture was then cooled to room temperature, diluted with 2-MeTHF and washed with saturated aqueous NaHCO ₃ and brine. The organic layer was separated, dried over sodium sulfate, filtered and concentrated in vacuo, and the residue was then purified by flash chromatography on C18 (MeCN/10 mM aqueous ammonium formate, pH 3.8, 0 to 100% gradient). The title compound was obtained as a light brown oil (58.0 mg, 0.10 mmol, 24% yield). Step 3 : (S)-7-( Dimethylphosphatyl )-3-(2-((5,5 -dimethylpiperidin -3 -yl ) amino )-5-( trifluoromethyl ) pyrimidin -4- yl )-1H- indole -6 -carbonitrile

To a stirred solution of ( S )-3-(2-((1-benzyl-5,5-dimethylpiperidin-3-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphonyl)-1H-indole-6-carbonitrile (58.0 mg, 0.10 mmol) in EtOH (12.5 mL) under nitrogen atmosphere, we added Pd/C 10% w/w (1.1 mg, 0.01 mmol) and _Boc2O (di-tert-butyl dicarbonate; 65.5 mg, 0.30 mmol). The reaction mixture was evacuated and backfilled with nitrogen (×3), followed by hydrogen. The reaction mixture was then stirred at room temperature under hydrogen atmosphere overnight. After 16 hours, we observed incomplete conversion and therefore the reaction mixture was filtered through a CELITE® pad and concentrated under reduced pressure. The reaction was then repeated with the residue as described above. After almost complete consumption of the starting material (48 hours), the reaction mixture was filtered through a CELITE® pad and concentrated in vacuo to give the crude product, which was carried on to the next step . Therefore, the resulting oil was redissolved in DCM (5 mL) and TFA (0.23 mL, 3.0 mmol) was added. The reaction mixture was stirred at room temperature overnight. The mixture was then concentrated in vacuo and the residue was purified by reverse phase chromatography over C18 (MeCN/10 mM aqueous ammonium formate, pH 3.8, 0-100% gradient) to give the title compound (11.11 mg, 0.023 mmol, 23% yield over two steps) as a white solid.

Example 7 : Inhibition of CDK kinase activity We analyzed several compounds for inhibition of CDK7, CDK9, CDK12, and CDK2 activity at Biortus Biosciences (Jiangyin, Jiangsu, China) using a kinase assay for each CDK cultured using a Caliper/LabChip EZ Reader (Perkin Elmer, Waltham, MA). These assays measure the amount of phosphorylated peptide substrate produced as a fraction of total peptide after an incubation period at 27°C with the following components: test compound (variable concentration in a series of 3-fold serial dilutions from 10 µM down to 0.508 nM), active CDK protein (with the indicated cyclin, listed below for each CDK), ATP ( K _m concentration listed below for each CDK/cyclin or 2 mM ATP), and substrate peptide (listed below) in the following buffer: 2-(N-morpholino)ethane sulfonate (MES buffer, 20 mM), pH 6.75, 0.01% (v/v) Tween 20 detergent, 0.05 mg/mL bovine serum albumin (BSA), and 2% DMSO.

Specifically, the CDK7 inhibition assay _was performed using the CDK7/cyclin H/MAT1 complex (6 nM) and the "5-FAM-CDK7tide" peptide substrate ( 2 µM, a synthetic fluorophore-labeled peptide with the sequence 5-FAM-YSPTSPSYSPTSPSYSPTSPSKKKK (SEQ ID NO:1), where "5-FAM" is 5-carboxyluciferin), which under these conditions targets CDK7/ The apparent ATP K _m of cyclin H/MAT1 is 50 µM. CDK9 inhibition assay was performed by buffer composition containing ₁₀ mM MgCl as listed above, using CDK9/cyclin T1 complex (8 nM) and "5-FAM-CDK9tide" peptide substrate (2 µM, with the following sequence Synthetic fluorophore-labeled peptide: 5-FAM-GSRTPMY-NH ₂ (SEQ ID NO: 2), where 5-FAM is as defined above and NH ₂ represents a C-terminal amide. CDK12 inhibition assay was performed by the above-mentioned method containing 2 Buffer composition of mM MgCl ₂ using CDK12 (aa686-1082)/cyclin K complex (50 nM) and "5-FAM-CDK9tide" (2 µM) as defined above. CDK2 inhibition assay was performed as above The listed buffer compositions containing 2 mM MgCl ₂ use CDK2/cyclin E1 complex (0.5 nM) and "5-FAM-CDK7tide" as defined above (2 µM).

The incubation period for each CDK inhibition assay at 27°C was chosen so that the fraction of phosphorylated peptide product generated in each assay relative to the total peptide concentration was approximately 20% (±5%) for the uninhibited kinase (35 minutes for CDK7, 35 minutes for CDK2, 3 hours for CDK12, and 15 minutes for CDK9). In the case where a compound titration was tested and resulted in inhibition of peptide product formation, these data were fitted to generate the best-fit _IC50 value. _The K value or _{apparent affinity of each inhibitor for each CDK/cyclin was calculated from the kinase activity inhibition assay using the best-fit IC50 value at KmATP for} each CDK/cyclin except CDK7/cyclin H/MAT1 (Cheng and Prusoff, Biochem. Pharmacol. , 22(23)3099-3108, 1973), corrected for inhibitor consumption due to enzyme concentration (Copeland, "Evaluation of Enzyme Inhibitors in Drug Disclover: A Guide for Medicinal Chemists and Pharmacologists", 2nd edition, March 2013; ISBN: 978-1-118-48813-3):

Due to the limitations of tight-binding inhibition and CDK7/cyclin H/MAT1 analyses, surface plasmon resonance (SPR) measurements as described below were used to measure K _or direct compound binding affinities rather than calculating the apparent value of each inhibitor. K _i value.

Example 8 : CDK7/ Cyclin H surface plasmon resonance (SPR) Analytical method. We measured the binding kinetics and affinity of selected compounds to CDK7/cyclin H dimer using a Biacore T200 surface plasmon resonance (SPR) instrument (GE Healthcare). The dimer was amine-coupled to a CM5 sensor chip at a concentration of 12.5 µg/mL in 10 mM MES buffer at pH 6.5 at a flow rate of 10 µL/min. The target protein was immobilized on two flow cells for 12-16 seconds to obtain an immobilized protein content of 200-400 reaction units.

Compounds were titrated from 10 mM HEPES buffer, pH 7.5, with 150 mM NaCl, 0.05% surfactant P20 and 0.0002% DMSO, in 9 steps of 2-fold serial dilutions from 0.08-20 nM. Each compound concentration cycle was performed at 100 µL/min with 10 mM glycine, pH 9.5, with a 70 s contact time, 300 s dissociation time, 60 s regeneration time and 400 s stabilization time. For each compound, 0 nM flow cell-bound compound control and reference were subtracted to remove background and normalize data. Compound titrations were fitted globally using kinetic mode by Biacore T200 evaluation software (GE Healthcare). The best fitting values for compound association rates ( k _on ) and dissociation rates ( k _off ) for CDK7/cyclin H were determined and used to calculate compound affinity ( K _d ) for CDK7/cyclin H using the following equation:

The selectivity of the compound for CDK7 over CDK2, CDK9 or CDK12 was based on the expression of the target CDKK _i Values relative to direct compound binding to CDK7K _d , measured by SPR according to the following equation: The inhibition and dissociation constants and selectivities of the indicated compounds (three compounds of the invention and four comparators) against CDK2, CDK7, CDK9 and CDK12 are shown in the table of Figure 1 . As can be seen, each of the compounds of the invention is at least 3-fold and up to 3,000-fold more specific for CDK7 than the other CDKs tested.

Examples 9 ：Inhibition of cell proliferation ( compound 100-102) . HCC70 cell lines are derived from human TNBC, and we tested the ability of representative compounds of the invention to inhibit the proliferation of these cells at varying concentrations (from 4 pF M to 126.4 pM; 0.5 log serial dilutions). More specifically, we tested the selectivity against CDK7 of the same compounds tested above (whose structures are shown in Figure 1), and we used the known CDK inhibitor dinaciclib (or N-( (1S,3R)-3-((5-chloro-4-(1H-indol-3-yl)pyrimidin-2-yl)amino)cyclohexyl)-5-((E)-4-(di Methylamino)but-2-enamide)picolinamide) and triptolide were used as positive controls. Cells were incubated at 37°C in 5% CO₂ The cells were grown in ATCC formulated RPMI-1640 medium (ATCC 30-2001) supplemented with 10% fetal bovine serum (FBS) in a humidified chamber in the presence of 5% paraformaldehyde. Proliferation assays were performed over a 72 hour period using the CyQUANT® Direct Cell Proliferation Assay (Life Technologies, Chicago, IL USA) according to the manufacturer's instructions and using reagents provided with the kit. The results of the assay are shown in the table below. compound HCC70 EC ₅₀ (nM) Compound 100 0.975726 Compound 101 5.451 Compound 102 2.06403 Comparator 1 0.531255 Comparator 2 255.677 Comparator 3 23.8169 Comparator 4 108.274

Example 10 : Tumor growth inhibition in patient-derived xenograft (PDX) model. To assess tumor growth inhibition in selected in vivo PDX models of estrogen receptor-positive breast cancer (ER+BC) that are resistant to or resistant to the CDK4/6 inhibitor palboheib (ST1799, n=1) Both palboceib and fulvestrant (ST941, n=1) are resistant. Start dosing when the tumor is 100-200 ^mm3 . Use Compound 101, QD (6 mg/kg once daily, orally); Fulvestrant, SC (2.5 mg/kg, once weekly, by subcutaneous injection); Palboheib, QD ( 50 mpk, once daily, orally), or with a combination of Compound 101 (6 mg/kg, once daily, orally) and fulvestrant (2.5 mg/kg, once weekly, by subcutaneous injection) Mice were treated for 28 days, followed by observation for 21 days. The tumor growth inhibition (TGI) was calculated using the following formula on the last day of administration: TGI=(V _c1 -V _t1 )/(V _c0 -V _t0 ), where V _c1 and V _t1 are the control and treatment groups at the time of tumor extraction The average volume of V _c0 and V _t0 is the same group at the beginning of dosing.

In the palbociclib-resistant ER+BC PDX (ST1799) model, the combination of Compound 101 and Fulvestrant induced a significant TGI (89%) with no apparent tumor regrowth up to 21 days after dosing was stopped, which differentiated the observed effects from those of Compound 101 (83%), Fulvestrant (60%), or palbociclib (21%) when administered as single agents. In addition, the combination of Compound 101 and Fulvestrant was superior to the standard of care combination of palbociclib and Fulvestrant (75%). In the palbociclib and Fulvestrant dual-resistant ER+BC PDX model (ST941), Compound 101 administered as a single agent caused a 33% TGI, and Fulvestrant and palbociclib were not active as single agents or in combination with Fulvestrant and palbociclib. In contrast, the combination of Compound 101 and Fulvestrant exhibited a significant TGI (68%; p<0.0001 compared to Fulvestrant as a single agent), indicating resensitization to Fulvestrant. Figure 2 illustrates TGI results from the palbociclib-resistant HR+BC PDX model ST1799, and Figure 3 illustrates TGI results from the palbociclib- and Fulvestrant-resistant HR+BC PDX model ST941. We also observed TGI in four additional PDX models; BR5010 (modeling TNBC), LU5178 (modeling small cell lung cancer (SCLC)), OV15398 (modeling high-grade serous ovarian cancer (HGSOC)), and ST390 (modeling pancreatic ductal adenocarcinoma (PDAC)). In the TNBC model, compound 101 was orally administered to tumor-bearing NOD/SCID mice at 10 mg/kg QD or 5 mg/kg BID for 21 days. In the SCLC and HGSOC models, compound 101 was orally administered to tumor-bearing NOD/SCID mice at 3 mg/kg BID for 21 days. In the PDAC model, compound 101 was orally administered to tumor-bearing NOD/SCID mice at 6 mg/kg QD. In the TNBC, SCLC, and HGSOC models, tumor volume was measured during the treatment period and for an additional 21 days after treatment cessation. The % TGI observed at the end of treatment (Day 21) was calculated as: 1-[(Average TV Compound 101@EOT-Average TV Compound 101@Day 0)/(Average TV Veh@EOT-Average TV Veh@Day 0)]×100. The % Regression was calculated as: (Average TV Compound 101@EOT)/(Average TV Compound 101@Day 0)×100. The same calculation was used for the end of the study (Day 42). The results are shown in Figure 4. These results demonstrate deep and sustained TGI, including regressions, at well-tolerated doses in multiple tumor types. Dose-dependent transcriptional responses in xenograft tissues were observed within 4 hours of dosing and persisted for 24 hours. Similar TGIs were seen when the same total dose was administered QD or BID in the TNBC PDX model, suggesting that the effect was AUC or C _min driven. Furthermore, the TGI observed in SCLC (LU5178 PDX model) was not observed in previous studies using covalent CDK7 inhibitors (data not shown). Regarding the PDAC model, we found that Compound 101 was able to induce 100% TGI over the time examined (approximately 28 days) at a dose maximum below the MTD: on day 21, tumor volume was approximately 1,250 mm ³ in vehicle-treated mice, but only approximately 250 mm ³ in Compound 1-treated mice (6 mg/kg QD, PO). While compound 101 (a non-covalent CDK7 inhibitor) could achieve 100% TGI at sub-MTD doses in the PDAC PDX tumors tested, the covalent CDK7 inhibitor achieved only modest TGI at its MTD (40 mg/kg BIW, by IV administration, with significant weight loss (8.4%) and necrosis at the injection site; data not shown).

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments described herein. The embodiments described in detail herein are not intended to limit the scope of the invention. Those skilled in the art will appreciate that various changes and modifications can be made to the described embodiments without departing from the spirit or scope of the invention as defined in the following claims.

Figure 1 is a table depicting the inhibition and dissociation constants and selectivities of the indicated compounds (three compounds of the invention and four comparators) against CDK2, CDK7, CDK9 and CDK12.

Figure 2 is a line graph depicting tumor volume (mm ³ ) over time (days) in the Palboceb anti-HR+BC PDX model ST1799 (as further described in the Examples below).

Figure 3 is a line graph depicting tumor volume ( ^mm3 ) as a function of time (days) in the pabboceb and fulvestrant anti-HR+BC PDX model ST941 (as further described in the Examples below).

Figure 4 is a line graph showing tumor volume (mm ³ ) as a function of time (days) in three PDX models depicting TNBC (BR5010; top), small cell lung cancer (LU5178; middle), and ovarian cancer (OV15398; bottom). picture. Animals were treated with compound 101 as described in Example 10. Data obtained from vehicle-treated (control) animals are represented by filled circles (upper trace in each figure). Data from animal models of TNBC given 10 mg/kg Compound 101 QD are represented by solid squares in the top panel; the dose of 5 mg/kg BID is represented by triangles. The triangles in the middle and bottom panels also represent data obtained from animal models of SCLC and ovarian cancer treated with Compound 101.

Claims (20)

A compound or a pharmaceutically acceptable salt, hydrate or stereoisomer thereof, wherein the compound has the structural formula (I):

wherein: R ¹ is methyl or ethyl; R ² is methyl or ethyl; R ³ is 5-methylpiperidin-3-yl, 5,5-dimethylpiperidin-3-yl, 6-methylpiperidin-3-yl or 6,6-dimethylpiperidin-3-yl, wherein one or more hydrogen atoms in R ³ are replaced by deuterium as appropriate; and R ⁴ is -CF ₃ or chlorine. For example, the compound of claim 1 or its pharmaceutically acceptable salt, hydrate or stereoisomer, the compound has structural formula (Ia):

, where R ³ is

The compound of claim 2 or its pharmaceutically acceptable salt, hydrate or stereoisomer, wherein the compound is:

The compound of claim 3 or its pharmaceutically acceptable salt, hydrate or stereoisomer, wherein the compound is

The compound of claim 1 or a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt thereof, a hydrate or a stereoisomer thereof, wherein one or more hydrogen atoms in R ³ are replaced by deuterium. For example, the compound of claim 1 or its pharmaceutically acceptable salt or its pharmaceutically acceptable salt, hydrate or stereoisomer, wherein none of the hydrogen atoms in ^R3 is replaced by deuterium. For example, the compound of claim 1 or its pharmaceutically acceptable salt, hydrate or stereoisomer, wherein the pharmaceutically acceptable salt is an acid addition salt. The compound of claim 7 or its pharmaceutically acceptable salt, hydrate or stereoisomer, wherein the acid addition salt is formed with lactic acid, hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, perchloric acid, acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid. A pharmaceutical composition comprising a compound as claimed in any one of claims 1 to 8 or a pharmaceutically acceptable salt, hydrate or stereoisomer thereof, and a pharmaceutically acceptable carrier. The pharmaceutical composition of claim 9, wherein the composition is formulated for oral administration. The pharmaceutical composition of claim 9, wherein the composition is formulated in unit dosage form. A use of a pharmaceutical composition as claimed in claim 9, which is used to prepare a drug for treating a disease, wherein the disease is a proliferative disease, an inflammatory disease, an autoinflammatory disease, an autoimmune disease or an infectious disease. The use of the pharmaceutical composition as claimed in claim 12, wherein the disease is a proliferative disease. The use of the pharmaceutical composition of claim 12, wherein the proliferative disease is cancer, benign neoplasm or pathological angiogenesis. The use of the pharmaceutical composition as claimed in claim 14, wherein the cancer is a blood cancer. Use of a pharmaceutical composition as claimed in claim 14, wherein the cancer is characterized by the presence of a solid tumor. A kit comprising the pharmaceutical composition of claim 9, instructions for use and optionally a second agent selected from the group consisting of anti-proliferative agents, anti-cancer agents, immunosuppressants and analgesics. A use of a pharmaceutical composition as claimed in claim 9 for preparing a medicament for treating a disease, wherein the disease is a proliferative disease, an inflammatory disease, an autoinflammatory disease, an autoimmune disease or an infectious disease in an individual who has been determined to have one or more of the following: elevated CDK7 expression or activity; high-grade cancer; a cellular phenotype with overexpression of steroid receptors; triple-negative breast cancer; and resistance to previously administered chemotherapy. The use as claimed in claim 18, wherein the previously administered chemotherapeutic agent is a CDK inhibitor or a steroid receptor degrader. The use of claim 19, wherein the CDK inhibitor is palbociclib and the steroid receptor degrader is fulvestrant.