RU2513636C2 - Some chemical structures, compositions and methods - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new compounds of structural formula I that possess inhibitory activity on phosphatidyl inositol-3-kinase (PI3-kinase). In formula

I: B represents group of formula

II:, wherein W_c represents 6-10-merous aryl, q is equal to 0, 1, 2, 3 or 4; X is absent or represents -(CH(R⁹))_z-, z is equal to 1; Y represents -N(R⁹)-; W_d represents

R¹ and R² represents C_1-6alkyl or halogeno; R³ represents hydrogen or C_1-6alkyl; and in each case R⁹ independently represents hydrogen or C_1-6alkyl.

EFFECT: invention refers to a pharmaceutical composition containing the above compounds, and to a method for P3I-kinase inhibition in a subject, wherein the subject suffers a disease representing cancer, a bone disorder, an inflammatory disease, an immune disease, a nervous system disease, a metabolic disease, a respiratory disease, thrombosis or a cardiovascular disease.

24 cl, 11 dwg, 6 tbl, 43 ex

Description

In this application, priority advantage is claimed in respect of provisional applications of the United States with serial No. 61/009971, filed January 4, 2008; 61/194294, filed September 26, 2008, and 61/201146, filed December 5, 2008, and at the same time pending application under the number 35280-714.602 in the register of patent attorneys, filed at the same time, each of which is thereby included in its entirety completeness by reference.

BACKGROUND OF THE INVENTION

Cell activity can be regulated by external signals that stimulate or inhibit intracellular events. The process by which stimulatory or inhibitory signals are transmitted to and within the cell to elicit an intracellular response is called signal transduction. Over the past decades, cascades of events occurring during signal transduction have been identified and found to play a central role in a variety of biological responses. Disorders in various components of signal transduction pathways have been found to cause a large number of diseases, including numerous forms of cancer, inflammatory disorders, metabolic disorders, vascular and nervous diseases (Gaestel et al. Current Medicinal Chemistry (2007) 14: 2214-2234).

Kinases are a class of important signaling molecules. Kinases in general can be subdivided into protein kinases and lipid kinases, and some kinases exhibit double specificity. Protein kinases are enzymes that phosphorylate other proteins and / or themselves (i.e., autophosphorylation). Protein kinases in general can be divided into three main groups based on their use of the substrate: tyrosine kinases, which mainly phosphorylate substrates on tyrosine residues (e.g. erb2, PDGF receptor (platelet growth factor), EGF receptor (epidermal growth factor), VEGF receptor (factor vascular endothelial growth), src, abl), serine / threonine kinases, which preferably phosphorylate substrates at serine and / or threonine residues (e.g. mTorC1 (Rapamycin target complex 1 in mammals), mTorC2, ATM (mutated ataxia-body gene ngiektazii), ATR (Ataxia-modified telangiectasias and related Rad3), DNA-PK (DNA-activated protein kinase), Akt (protein kinase B)) and dual specificity kinases which phosphorylate substrates on tyrosine residues, serine and / or threonine.

Lipid kinases are enzymes that catalyze the phosphorylation of lipids. These enzymes and the resulting phosphorylated lipids and biologically active organic molecules derived from lipids play a role in many diverse physiological processes, including cell proliferation, migration, adhesion and differentiation. Some lipid kinases are membrane bound and catalyze the phosphorylation of lipids contained in cell membranes or bound to cell membranes. Examples of such enzymes include phosphoinositide kinases (such as PI3 (phosphatidylinositol-3) kinases, PI4 kinases), diacylglycerol kinases and sphingosine kinases.

The signaling pathway involving phosphoinositide-3-kinases (PI3K) is one of the systems that are most susceptible to mutations in various types of cancer in humans. Signal transmission through PI3K is also a key factor in many other human diseases. The PI3K signaling process is involved in many painful conditions, including allergic contact dermatitis, rheumatoid arthritis, osteoarthritis, inflammatory bowel disease, chronic obstructive pulmonary disorder, psoriasis, multiple sclerosis, asthma, disorders associated with diabetes complications and inflammatory complications of the cardiovascular system such as acute coronary syndrome.

PI3K are members of a unique and conservative family of intracellular lipid kinases that phosphorylate the 3'-OH group of phosphatidylinositol or phosphoinositide. The PI3K family contains 15 kinases with various substrate specificities, expression patterns, and regulatory methods (Katso et al., 2001). Class I PI3K (p110α, p110β, p110δ and p110γ) are usually activated by tyrosine kinases or G-protein-coupled receptors to produce PIP3 (phosphatidylinositol triphosphate), which involves downstream effectors such as effectors in the Akt / PDK1 pathway ( Akt / phosphoinositide-dependent kinase-1), mTOR, kinases of the Tec family and GTPases of the Rho family. Class II and III PI3 kinases play a key role in intracellular signal transmission through the synthesis of PI (3) P (phosphatidylinositol-3-phosphate) and PI (3,4) P2 (phosphatidylinositol-3,4-bisphosphate). PIKKs (phosphatidylinositol kinases) are protein kinases that regulate cell growth (mTORCI) or control genome integrity (ATM, ATR, DNA-PK and hSmg-1).

The isoform delta (δ) PI3K class I is involved, in particular, in a number of diseases and biological processes. PI3Kδ is expressed mainly in hematopoietic cells, including leukocytes such as T cells, dendritic cells, neutrophils, mast cells, B cells and macrophages. PI3Kδ is involved as an integral part of mammalian immune system functions such as T-cell function, B-cell activation, mast cell activation, dendritic cell function and neutrophil activity. Due to its inherent role in the functioning of the immune system, PI3Kδ is also involved in a number of diseases associated with an undesirable immune response, such as allergic reactions, inflammatory diseases, inflammatory mediated angiogenesis, rheumatoid arthritis, autoimmune diseases such as lupus erythematosus, asthma, emphysema and others respiratory diseases. Another Class I PI3K involved in the functioning of the immune system includes PI3Kγ, which plays a role in leukocyte signaling and is involved in inflammation, rheumatoid arthritis, and autoimmune diseases such as lupus erythematosus.

Mediators located downstream of the PI3K signal transduction pathway include Akt and mammalian rapamycin target (mTOR). Akt contains a plextrin homologous (PH) domain that binds to PIP3, which causes activation of Akt kinase. Akt phosphorylates many substrates and is the central downstream PI3K effector to produce a variety of cellular responses. One of the important functions of Akt is to increase the activity of mTOR by phosphorylation of TPPs (tuberous sclerosis complex) 2 and other mechanisms. mTOR is a serine-threonine kinase related to the lipid kinases of the PI3K family. mTOR is involved in a wide range of biological processes, including cell growth, cell proliferation, motility and cell survival. Dysregulation of the mTOR pathway has been reported in various types of cancer. mTOR is a multifunctional kinase that integrates signals from growth factors and nutrients to regulate protein translation, nutrient intake, autophagy and mitochondrial function.

In essence, kinases, in particular PI3K, are critical targets for drug development. There remains a need for PI3K inhibitors suitable for drug development. The present invention addresses this need and provides related benefits, including by offering new classes of kinase inhibitors.

SUMMARY OF THE INVENTION

In one aspect of the present invention, the following compounds of formula I or their pharmaceutically acceptable salts are provided, wherein

W _d represents heterocycloalkyl, aryl or heteroaryl;

B is a grouping of formula II:

where W _c represents aryl, heteroaryl, heterocycloalkyl or cycloalkyl, and q is an integer of 0, 1, 2, 3 or 4;

X is absent or represents - (CH (R ⁹ )) _z -, and z is an integer of 1;

Y is absent or is —N (R ⁹ ) -;

R ¹ represents hydrogen, alkyl, alkenyl, alkynyl, alkoxy, amido, alkoxycarbonyl, sulfonamido, halo, cyano or nitro;

R ² represents alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, heteroarylalkyl, alkoxy, amino, halo, cyano, hydroxy or nitro;

R ³ represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy, amido, amino, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy or nitro;

R ⁵ , R ⁶ , R ⁷ and R ⁸ independently represent hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy, amido, amino, acyl, acyloxy, sulfonamido, halo, cyano, hydroxy or nitro; and

in each case, R ⁹ independently represents hydrogen, alkyl, cycloalkyl or heterocycloalkyl.

In another aspect of the present invention, there are provided compounds comprising compounds of formula I or pharmaceutically acceptable salts thereof, wherein

W _d represents heterocycloalkyl, aryl or heteroaryl;

B represents an alkyl, amino, heteroalkyl or moiety of formula II:

,

,

where W _c represents aryl, heteroaryl, heterocycloalkyl or cycloalkyl, and q is an integer of 0, 1, 2, 3 or 4;

X is absent or represents - (CH (R ⁹ )) _z - and z independently represents an integer of 1, 2, 3 or 4;

Y is absent, represents -O-, -S-, -S (= O) -, -S (= O) ₂ -, -N (R ⁹ ) -, -C (= O) - (CHR ⁹ ) ₂ - -C (= O) -, -N (R ⁹ ) (C = O) -, -N (R ⁹ ) (C = O) NH - or -N (R ⁹ ) C (R ⁹ ) ₂ -;

R ¹ represents hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, hydroxy urea or carbonate;

R ² represents alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro, hydroxy, nitro, nitro, hydroxy, nitro carbonate;

R ³ represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, aryl, heteroaryl or nitro;

R ⁵ , R ⁶ , R ⁷ and R ⁸ independently represent hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₅ alkenyl, C ₂ -C ₅ alkynyl, C ₃ -C ₅ cycloalkyl, C ₁ -C ₄ heteroalkyl C ₁ -C ₄ alkoxy, C ₁ -C ₄ amido, amino, acyl, C ₁ -C ₄ acyloxy, alkoxycarbonyl, C _1- C ₄ sulfonamido, halo, cyano, hydroxy or nitro; and

in each case, R ⁹ independently represents hydrogen, C ₁ -C ₁₀ alkyl, C ₃ -C ₇ cycloalkyl, heterocycloalkyl or C ₂ -C ₁₀ heteroalkyl.

In some of the embodiments, X is —CH ₂ -, —CH (CH ₂ CH ₃ ), or —CH (CH ₃ ) -.

In some embodiments, XY is —CH ₂ —N (CH ₃ ), —CH ₂ —N (CH ₂ CH ₃ ), —CH (CH ₂ CH ₃ ) —NH—, or —CH (CH ₃ ) —NH—.

In some embodiments, W _d is pyrazolopyrimidine of formula III:

,

,

where R ¹¹ represents H, alkyl, halo, amino, amido, hydroxy or alkoxy, and R ¹² represents H, alkyl, alkynyl, alkenyl, halogen, aryl, heteroaryl, heterocycloalkyl or cycloalkyl. In some embodiments, W _d is a pyrazolopyrimidine of formula III, wherein R ¹¹ is H, alkyl, halo, amino, amido, hydroxy or alkoxy, and R ¹² is cyano, amino, carboxylic acid, or amido.

In some embodiments, the compound of formula I has the structure of formula IV:

,

,

where R ¹¹ represents H, alkyl, halo, amino, amido, hydroxy or alkoxy, and R ¹² represents H, alkyl, alkynyl, alkenyl, halogen, aryl, heteroaryl, heterocycloalkyl or cycloalkyl. In some embodiments, the compound of formula I has the structure of formula IV, wherein R ¹¹ is H, alkyl, halo, amino, amido, hydroxy or alkoxy, and R ¹² is cyano, amino, carboxylic acid, or amido.

In some embodiments of a compound of formula I, IR ¹¹ is amino. In some embodiments of a compound of formula I, IR ¹² is alkyl, alkenyl, alkynyl, heteroaryl, aryl, or heterocycloalkyl. In some embodiments of the compounds of formula IR ¹² is cyano, amino, carboxylic acid or amido. In some embodiments of the compounds of formula IR ¹² is monocyclic heteroaryl. In some embodiments of the compounds of formula IR ¹² is a bicyclic heteroaryl.

In some embodiments of a compound of formula I, this compound has the structure of formula V:

.

.

In some of the embodiments of formula I, XY represents N (CH ₂ CH ₃ ) CH ₂ - or N (CH ₃ ) CH ₂ -. In some of the embodiments of formula I, the compound has the structure of formula VI:

In some of the embodiments of the formula, IR ³ is —H, —CH ₃ ) —Cl, or —F.

In some of the embodiments of the formula, IR ⁵ , R ⁶ , R ⁷ and R ⁸ are hydrogen.

In some of the embodiments of formula I, B is a grouping of formula II:

,

,

where W _c represents aryl, heteroaryl, heterocycloalkyl or cycloalkyl, and q is an integer of 0, 1, 2, 3 or 4.

In another aspect of the invention, there is provided a compound having the structure of formula I, and pharmaceutically acceptable salts thereof, wherein B is a moiety of formula II,

where W _c represents aryl, heteroaryl, heterocycloalkyl or cycloalkyl; and q is an integer of 0, 1, 2, 3, or 4;

X is absent or represents - (CH (R ⁹ )) _z -; and z is independently an integer of 1;

Y is absent or is —N (R ⁹ ) -;

, или когда Y присутствует, W_d представляет собой:

;when Y is absent, W _d represents:

or when Y is present, W _d is:

;

R ¹ represents hydrogen, alkyl, alkenyl, alkynyl, alkoxy, amido, alkoxycarbonyl, sulfonamido, halo, cyano or nitro;

R ² represents alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, heteroarylalkyl, alkoxy, amino, halo, cyano, hydroxy or nitro;

R ³ represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy, amido, amino, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy or nitro;

in each case, R ⁹ independently represents hydrogen, C ₁ -C ₁₀ alkyl, cycloalkyl or heterocycloalkyl; and R ¹² represents H, alkyl, alkynyl, alkenyl, halo, aryl, heteroaryl, heterocycloalkyl or cycloalkyl.

In another aspect of the invention, there is provided a compound having the structure of Formula I, and pharmaceutically acceptable salts thereof, wherein:

B represents an alkyl, amino, heteroalkyl or moiety of formula II,

where W _c represents aryl, heteroaryl, heterocycloalkyl or cycloalkyl; and q is an integer of 0, 1, 2, 3, or 4;

X is absent or represents - (CH (R ⁹ )) _z -; and z independently represents an integer of 1, 2, 3 or 4;

Y is absent, represents —N (R ⁹ ) - or —N (R ⁹ ) —CH (R ⁹ ) -;

или

;W _d represents:

or

;

R ¹ represents hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, hydroxy urea or carbonate;

R ² represents alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro;

R ³ represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro, aryl or heteroaryl;

in each case, R ⁹ independently represents hydrogen, C ₁ -C ₁₀ alkyl, C ₃ -C ₇ cycloalkyl, heterocycloalkyl or C ₂ -C ₁₀ heteroalkyl; and

R ¹² represents H, alkyl, alkynyl, alkenyl, halo, aryl, heteroaryl, heterocycloalkyl, cycloalkyl, cyano, amino, carboxylic acid, alkoxycarbonyl or amido.

In some embodiments, R ³ is —H, —CH ₃ , —CH ₂ CH ₃ , —CF ₃ , —Cl, or —F. In other embodiments, R ³ is —CH ₃ , —Cl, or —F.

In some embodiments, B is a moiety of formula II, wherein W _c is aryl, heteroaryl, heterocycloalkyl or cycloalkyl; and q is an integer of 0, 1, 2, 3, or 4. In some embodiments, B is heterocycloalkyl. In some embodiments, B is monocyclic heteroaryl. In some embodiments, B is substituted phenyl. In some embodiments, B is substituted alkyl.

In some embodiments, B is - (CH ₂ ) ₂ —NR ^a R ^a , where each R ^a independently is hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl, or —NR ^a R ^{a are} joined together to form a cyclic moiety.

In some embodiments, R ¹ is H, —F, —Cl, —CN, —CH ₃ , isopropyl, —CF ₃ , —OCH ₃ , nitro, or phosphate. In some embodiments, R ¹ is phosphate.

In some embodiments, R ² represents halogen, hydroxy, cyano or nitro; and q is 1. In some embodiments, R ² is phosphate; and q is 1. In some embodiments, R ² is alkyl or halogen; and q is 1 or 2.

In some embodiments, X is —CH ₂ -, —CH (CH ₂ CH ₃ ), or —CH (CH ₃ ) -. In some embodiments, XY is —CH ₂ —N (CH ₃ ), —CH ₂ —N (CH ₂ CH ₃ ), —CH (CH ₂ CH ₃ ) —NH—, or —CH (CH ₃ ) —NH—. In some embodiments, is N (CH ₂ CH ₃ ) CH ₂ - or N (CH ₃ ) CH ₂ -.

In some embodiments, R ¹² is monocyclic heteroaryl. In some embodiments, R ¹² is a bicyclic heteroaryl. In some embodiments, R ¹² is heterocycloalkyl.

In some embodiments, the compound of formula I has the structure of formula IV-A:

.

.

In some embodiments, the compound of formula I has the structure of formula V-A:

.

.

In some embodiments, the compound of formula I has the structure of formula IV-A or formula V-A.

In some embodiments, the compound of formula I has the structure of formula V-B:

.

.

In some embodiments, the compound of formula I has the structure of formula VI-A:

.

.

In another aspect of the invention, there is provided a composition comprising a pharmaceutically acceptable excipient and one or more compounds of formula I, formulas IV, IV-A, V, V-A, V-B, VI and VI-A. In some embodiments, the composition is in liquid, solid, semi-solid form, gel form, or aerosol form.

In another aspect of the invention, there is provided a method of inhibiting phosphatidylinositol-3-kinase (PI3 kinase), comprising contacting the PI3 kinase with an effective amount of one or more of the compounds described herein. For example, the contacting step involves the use of one or more compounds of the formula I, IV, IV-A, V, VA, VB, VI and VI-A. In some embodiments, the contacting step comprises contacting with a cell that contains said PI3 kinase. In some embodiments of the method, inhibition occurs in a subject suffering from a disorder associated with malfunctioning of one or more types of PI3 kinase. Some typical diseases involving the malfunctioning of one or more types of PI3 kinase are selected from the group consisting of autoimmune diseases, rheumatoid arthritis, respiratory disease, allergic reactions, and various types of cancer. If desired, the compound used in this method has the structure of formula IV, where R ¹¹ is amino, R ¹² is substituted phenyl.

In some embodiments of this method, the inhibition occurs in a subject suffering from rheumatoid arthritis or a respiratory disease, and where the compound has the structure of formula IV, where R ¹¹ is amino, R ¹² is bicyclic heteroaryl.

In some embodiments, the method comprises administering to the subject a second therapeutic agent.

In yet another aspect of the present invention, a method for treating a disease exhibiting an undesired immune response is provided. The method includes the step of administering to a subject in need of one or more compounds set forth in this application, including compounds of formula I, IV, IV-A, V, VA, VB, VI and / or VI-A in an amount that is effective in improving said unwanted immune response. In some embodiments, one or more compounds inhibit T cell-independent B cell activation, as evidenced by a decrease of at least about five-fold in the production of anti-TMP (trinitrophenyl) IgG3 when administered to the test animal at a dose of 2 times per day (BID ) in an amount of less than about 30 mg / kg.

In some embodiments, the disease to be treated is associated with swelling of a joint or joint pain in a subject. This method can be effective in improving one or more symptoms of rheumatoid arthritis, as evidenced by a decrease in the average diameter of the joint by at least about 10% after 17 days and / or by reducing the diameter of the ankle by at least 5-10% or more after treatment within a few days to several weeks, including, for example, a decrease in the diameter of the ankle joint by at least 5% after 7 days of treatment. In another embodiment, an unwanted immune response is confirmed by enhanced production of antibodies against type II collagen, and the use of one or more compounds of the invention reduces the level of antibodies against type II collagen in the blood serum with an ED50 of less than about 10 mg / kg.

Link inclusion

All publications, patents, and patent applications mentioned in this application are incorporated into this application by reference to the same extent as if it were specifically and individually indicated that each individual publication, patent, or patent application is incorporated by reference.

A brief description of the graphic materials

The accompanying claims set forth in detail the new features of the invention. A more complete understanding of the features and advantages of the present invention will be achieved by reference to the following detailed description, which sets forth illustrative embodiments, which use the principles of the invention, and in the accompanying graphic materials of which:

figure 1 shows a typical protocol for measuring T-cell independent production of TNP-specific antibodies in vivo (BDA1 - type A1 brachidactyly; ELISA - enzyme-linked immunosorbent assay);

figure 2 shows a multiple decrease in the magnitude of the TNP-specific IgGS response to antigens (Ag), due to compounds 7 and 53 of formula IV, compared with the solvent as a control when administered orally (p / o);

figure 3 shows the dose-dependent effect of the oral administration of compound 53 of formula IV twice a day to reduce the increase in the diameter of the ankle joint (Ank) over time in a model of developing collagen-induced arthritis in rats. The results are also presented for control rats without arthritis, control rats with arthritis, which were injected with a solvent as a negative control, and control rats with arthritis, which were administered methotrexate (MTX) twice a day (IK-1 - induced by collagen 1);

figure 4 shows the dose-dependent effect of compounds 7 and 53 of formula IV on the improvement of the histopathological picture of the ankle joint when introduced in the model of developing collagen-induced arthritis in rats. The results are also presented for control rats with arthritis, which were injected with a solvent as a negative control or methotrexate;

figure 5 shows the dose-dependent effect of compounds 7 and 53 of formula IV on improving the histopathological picture of the knee joint when rats are introduced into the model of developing collagen-induced arthritis. The results are also presented for control rats with arthritis, which were injected with a solvent as a negative control or methotrexate as a positive control;

figure 6 shows the dose-dependent effect of compounds 7 and 53 of formula IV on the decrease in the level of antibodies against type II collagen in vivo when introduced into the model of developing collagen-induced arthritis in rats. Also presented are the results for rats with arthritis that were given a solvent as a negative control or methotrexate (MTX);

7 shows the dose-dependent effect of compound 7 of formula IV on improving the histopathological picture of the ankle joint when rats are introduced into the model of developing collagen-induced arthritis. The results are also presented for control rats with arthritis that were injected with a solvent, and rats with arthritis that were injected with methotrexate;

on Fig depicts the dose-dependent effect of compound 53 of the formula IV, administered once a day, on the histopathology of the ankle joint in rats with a model of collagen-induced arthritis. The results are also presented for control rats with arthritis injected with a solvent, and rats with arthritis injected with enbrel;

figure 9 shows the dose-dependent effect of compound 53 of the formula IV, administered twice a day, on the histopathology of the ankle joint in a rat model of collagen-induced arthritis. The results are also presented for control rats with arthritis injected with a solvent, and rats with arthritis injected with enbrel;

figure 10 shows the dose-dependent effect of compound 53 of formula IV on the increase in average paw volume in a model of adjuvant-induced arthritis;

11 depicts the time effect of compound 53 of formula IV on the average rat weight in a rat adjuvant-induced arthritis model.

DETAILED DESCRIPTION OF THE INVENTION

Although preferred embodiments of the present invention are shown and set forth herein, it will be apparent to those skilled in the art that such embodiments are given by way of example only. Specialists in the art will immediately have the ideas of numerous variations, changes and substitutions without deviating from the invention. It should be understood that in the practical application of the invention, various alternatives to the embodiments of the invention set forth in this application may be used. It is intended that the appended claims define the scope of the invention and thereby cover methods and structures falling within the scope of this claims and their equivalents.

Unless otherwise indicated, all technical and scientific terms used in this application have the same meanings as they are commonly understood by a person skilled in the technical field to which this invention belongs. All patents and publications referred to in this application are incorporated by reference.

As used in the description and claims, the singular includes references to the plural, unless the context clearly dictates otherwise.

As used herein, “agent” or “biologically active agent” refers to a biological, pharmaceutical or chemical compound or other grouping. Non-limiting examples include a simple or complex organic or inorganic molecule, peptide, protein, oligonucleotide, antibody, antibody derivative, antibody fragment, vitamin derivative, carbohydrate, toxin, or chemotherapeutic compound. Various compounds can be synthesized, for example, small molecules and oligomers (e.g., oligopeptides and oligonucleotides) and synthetic organic compounds based on various core structures. Additionally, the compounds for screening can be obtained from various natural sources, such as, for example, extracts from plants or animals and the like. One skilled in the art can readily recognize that there is no limitation on the structural nature of the agents of the present invention.

The term “agonist,” as used herein, refers to a compound having the ability to initiate or enhance the biological function of a target protein, whether by inhibiting the activity or expression of a target protein. Accordingly, the term “agonist” is defined in the context of the biological role of the target polypeptide. Although the preferred agonists provided herein specifically interact with (e.g. bind to) the target, compounds that initiate or enhance the biological activity of the target polypeptide by interacting with other members of the signal transduction pathway, of which this polypeptide is a member The target is also specifically included within the scope of this definition.

The terms “antagonist” and “inhibitor” are used interchangeably and refer to a compound having the ability to inhibit the biological function of a target protein, whether by inhibiting the activity or expression of a target protein. Accordingly, the terms “antagonist” and “inhibitors” are defined in the context of the biological role of the target protein. Despite the fact that the preferred antagonists given in this application specifically interact with (for example, bind to) a target, compounds that inhibit the biological activity of a target protein by interacting with other members of the signal transduction pathway, one of which is this target protein are also specifically included in the scope of this definition. A preferred biological activity inhibited by an antagonist is associated with the development, growth or spread of a tumor, or an undesired immune response that manifests itself as an autoimmune disease.

An “anticancer agent”, “an antitumor agent” or a “chemotherapeutic agent” refers to any agent useful in the treatment of a neoplastic condition. One class of anticancer agents contains chemotherapeutic agents. "Chemotherapy" means the administration of one or more chemotherapeutic drugs and / or other agents to a patient suffering from cancer, in various ways, including intravenous, oral, intramuscular, intraperitoneal (ip), intravesical, subcutaneous (s / c), transdermal, buccal or inhaled or in the form of a suppository.

The term "cell proliferation" refers to a phenomenon due to which the number of cells changes as a result of division. This term also encompasses cell growth, as a result of which the morphology of cells changes (for example, they increase in size), which is consistent with a proliferative signal.

The term "co-administration", "administered in combination with" and their grammatical equivalents, as used herein, encompasses the administration of two or more agents to an animal so that both agents and / or their metabolites are present in the animal in the same time. Co-administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in compositions in which both agents are present.

The term "effective amount" or "therapeutically effective amount" refers to such an amount of the compound set forth in this application that is sufficient to obtain the effect of the intended use, including, but not limited to, the treatment of the diseases described below. The therapeutically effective amount may vary depending on the intended use (in vitro or in vivo) or the subject and the disease state that is treated, for example, the weight and age of the subject, the severity of the disease state, route of administration and the like, which can easily be determined by a person of ordinary skill in the art. The term also applies to a dose that will induce a specific response in target cells, for example, a decrease in platelet adhesion and / or cell migration. The specific dose will vary depending on the particular compound selected, the administration regimen to be followed, whether it is administered in combination with other compounds, timing of administration, the tissue into which it is administered, and the physical delivery system into which it is administered.

As used herein, the terms “treatment” or “treatment process” or “relief” or “decrease in intensity” are used interchangeably in this application. These terms refer to an approach for obtaining useful or desired results, including, but not limited to, therapeutic benefits and / or preventive benefits. The therapeutic benefit is understood to mean eliminating or reducing the intensity of the underlying disorder that is being treated. Also, therapeutic benefits are achieved by eliminating or decreasing the intensity of one or more physiological symptoms associated with the underlying disorder, so that the patient has an improvement, although the patient may still be affected by the underlying disorder. For prophylactic benefit, the compositions may be administered to a patient at risk of developing a particular disease, or to a patient with confirmed one or more physiological symptoms of the disease, possibly even if the diagnosis of this disease may not be made.

A “therapeutic effect”, as the term is used herein, encompasses a therapeutic benefit and / or a prophylactic benefit as described above. A preventive effect includes delaying or eliminating the occurrence of a disease or condition, delaying or eliminating an attack of symptoms of a disease or condition, slowing, stopping or reversing the development of a disease or condition, or any combination thereof.

The term “pharmaceutically acceptable salt” refers to salts derived from a number of organic and inorganic counterions well known in the art. Pharmaceutically acceptable acid addition salts may be formed with inorganic acids and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamon acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like. Pharmaceutically acceptable base addition salts may be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum and the like bases. Organic bases from which salts can be derived include, for example, primary, secondary and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, basic ion exchange resins and the like, specifically such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salt is selected from ammonium, potassium, sodium, calcium, and magnesium salts.

A "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes any and all solvents, dispersing media, coating agents, antibacterial and antifungal agents, isotonic and delaying absorption agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Unless any conventional medium or agent is incompatible with the active ingredient, their use in the therapeutic compositions of the invention is contemplated. Additional active ingredients may also be included in the composition.

"Signal transduction" is a process in which stimulating or inhibitory signals are transmitted to and within the cell in order to elicit an intracellular response. A signal transduction path modulator refers to a compound that modulates the activity of one or more cellular proteins mapped to the same specific signal transduction path. The modulator can increase (agonist) or suppress (antagonist) the activity of the signal molecule.

The term "selective inhibition" or "selectively inhibits", as applied to a biologically active agent, refers to the ability of an agent to selectively reduce a signal transmission target activity compared to a non-target signal transmission activity by directly or indirectly interacting with the target.

The term "B-ALL", which is used in this application, refers to B-cell acute lymphoblastic leukemia.

“Subject” refers to an animal, such as a mammal, such as a human. The methods described in this application may be useful both for human therapy and for applications in veterinary medicine. In some embodiments, the patient is a mammal, and in some embodiments, the patient is a human.

“Radiation therapy” means treating a patient using generally accepted methods and compositions known to a medical practitioner with radioactive emitters, such as alpha-emitting radionuclides (for example, actinium and thorium radionuclides), low linear energy transfer (LET) emitters (that there are beta emitters), conversion electron emitters (e.g. strontium-89 and samarium-153-EDTMP (samarium-153-ethylenediaminetetramethylene phosphonate)), or high-energy radiation, including, without limitation, x-rays , gamma rays and neutrons.

"Prodrug" is intended to indicate a compound that can be converted under physiological conditions or as a result of solvolysis into a biologically active compound described in this application. Thus, the term "prodrug" refers to a precursor of a biologically active compound that is pharmaceutically acceptable. A prodrug may be inactive when administered to a subject, but may be converted in vivo to an active compound, for example, by hydrolysis. A prodrug compound often offers advantages in solubility, tissue compatibility, or sustained release in a mammal (see, e.g., Bundgard, N., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam )). A discussion of prodrugs is provided in Higuchi, T., et al., "Pro-drugs as Novel Delivery Systems", A.C.S. Symposium Series, Vol. 14 and Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987; both are fully incorporated into this application by reference. The term "prodrug" is also intended to include any covalently linked carriers from which the active compound is released in vivo when such a prodrug is administered to a mammalian subject. Prodrugs of the active compound as described herein can be prepared by modifying the functional groups present in the active compound so that these modified compounds are cleaved either by standard manipulation or in vivo to give the starting active compound. Prodrugs include compounds where a hydroxy, amino or mercapto group is linked to any group that, when this prodrug of the active compound is administered to a mammal, is cleaved to form a free hydroxy group, a free amino group or a free mercapto group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of alcohol or acetamide, formamide and benzamide derivatives of an amine functional group in the active compound, and the like.

The term "in vivo" refers to an event that occurs in the body of a subject.

The term "in vitro" refers to an event that occurs outside the body of the subject. For example, an in vitro assay covers any assay conducted outside the subject. In vitro assays encompass cell assays using live or dead cells. In vitro assays also encompass cell-free assays in which no intact cells are used.

Also, unless otherwise indicated, it is understood that the structures described in this application include compounds that differ only in the presence of one or more isotope enriched atoms. For example, compounds having the presented structures, with the exception of replacing hydrogen with deuterium or tritium, or replacing carbon with ¹³ C or ¹⁴ C enriched carbon, are included in the scope of this invention.

Compounds of the present invention may also contain isotopes of atoms for one or more atoms that are part of such compounds in unnatural proportions. For example, compounds can be radioactively labeled with radioactive isotopes, such as, for example, tritium ( ³ N), iodine-125 ( ¹²⁵ I) or carbon-14 ( ¹⁴ C). All isotopic variations of the compounds of the present invention, both radioactive and non-radioactive, fall within the scope of the present invention.

In the case where ranges are used in this application to describe physical properties, such as molecular weight, or chemical properties, such as chemical formulas, it is understood that all combinations and subcombinations of ranges and the specific corresponding embodiments are included. The term “about” when it refers to a quantity or range of numerical values means that the quantity or range of numerical values referred to is an approximation within the range of experimental data (or within the statistical error of the experiment), and therefore the number or range of numerical values may vary for example, 1% -15% of the set quantity or range of numerical values. The term “comprising” (and related terms such as “comprising” or “comprising” or “having” or “including”) includes these embodiments, for example, an embodiment of any composition of an object, composition, method or process or the like, which “consist of "or" essentially consist of the "described features.

The following abbreviations and terms have the indicated meanings throughout this application:

PI3-K - phosphoinositide-3-kinase; PI - phosphatidylinositol; PDK - phosphoinositide-dependent kinase; DNA-PK - deoxyribonucleic acid dependent protein kinase; PTEN - deletion of phosphatase and tensin homolog on chromosome ten; PIKK - a kinase similar to phosphoinositide kinase; AIDS - acquired immunodeficiency syndrome; HIV - human immunodeficiency virus; Mel is methyl iodide; POCI3 - phosphorus oxychloride; KCNS - potassium isothiocyanate; TLC - thin layer chromatography; MeOH - methanol; and CHCl ₃ — chloroform.

The abbreviations used in this application have their traditional meaning in the field of chemistry and biology.

“Alkyl” refers to a straight or branched chain hydrocarbon radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, and having one to ten carbon atoms (eg, C ₁ -C ₁₀ alkyl). In all cases, when it appears in this application, a range of numerical values, such as "1-10", refers to each integer in a given range; for example, “1-10 carbon atoms” means that the alkyl group may consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms, although the present definition also covers a variant of the term “ alkyl "when no range of numerical values is indicated. In some embodiments, it is a C ₁ -C ₄ alkyl group. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, w-butyl, isobutyl, tertiary butyl, pentyl, isopentyl, neopentyl, hexyl, septyl, octyl, nonyl, decyl etc. Alkyl is attached to the remainder of the molecule via a single bond, for example methyl (Me), ethyl (Et), n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (mpem-butyl), 3-methylhexyl, 2-methylhexyl and the like. Unless specifically indicated otherwise in this description, the alkyl group is optionally substituted with one or more substituents, which independently are: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halogen, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR ^a , -SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -OC (O) N (R ^a ) ₂ , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N (R ^a ) C (O) N (R ^a ) ₂ , N (R ^a ) C (NR ^a ) N (R ^a ) ₂ , -N (R ^a ) S (O), R ^a (where t is 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a ) ₂ , where each R ^a independently represents hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Alkylaryl” refers to a - (alkyl) aryl radical, wherein aryl and alkyl are as disclosed herein and which are optionally substituted with one or more of one of the substituents described as suitable substituents for aryl and alkyl, respectively.

“Alkyl-vegetarian” refers to a - (alkyl) hetaryl radical, wherein hetaryl and alkyl are as described herein and which are optionally substituted with one or more of one of the substituents described as suitable substituents for aryl and alkyl, respectively.

“Alkylheterocycloalkyl” refers to a - (alkyl) heterocycle alkyl radical, wherein alkyl and heterocycloalkyl are as defined herein and which are optionally substituted with one or more of one of the substituents described as suitable substituents for heterocycloalkyl and alkyl, respectively.

An “alkene” group refers to a group consisting of at least two carbon atoms and at least one carbon-carbon double bond, and an “alkyne” group refers to a group consisting of at least two carbon atoms and at least one carbon -carbon triple bond. An alkyl group, whether it is saturated or unsaturated, may be a branched, straight chain or be cyclic.

"Alkenyl" refers to a straight or branched chain hydrocarbon radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond and having from two to ten carbon atoms (i.e., C ₂ -C ₁₀ alkenyl). In all cases, when it appears in this application, a range of numerical values such as "2-10" refers to each integer in a given range; for example, “2-10 carbon atoms” means that an alkenyl group may consist of 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms. In some embodiments, alkenyl contains from two to eight carbon atoms. In other embodiments, alkenyl contains from two to five carbon atoms (e.g., C ₂ -C ₅ alkenyl). Alkenyl is attached to the remainder of the molecule via a single bond, for example, ethenyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl, penta-1,4-dienyl, and the like. . Unless specifically indicated otherwise in this description, the alkenyl group is optionally substituted with one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl , trifluoromethoxy, nitro, trimethylsilanyl, -OR ^a , -SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -OC (O) N (R ^a ) ₂ , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N (R ^a ) C (O) N (R ^a ) ₂ , N (R ^a ) C (NR ^a ) N (R ^a ) ₂ , -N (R ^a ) S (O), R ^a (where t is 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a ) ₂ where each R ^a independently represents hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

"Alkenyl-cycloalkyl" refers to a - (alkenyl) cycloalkyl radical, wherein alkenyl and cycloalkyl are as described herein and which are optionally substituted with one or more of the substituents described as suitable substituents for alkenyl and cycloalkyl, respectively .

“Alkynyl” refers to a straight or branched chain hydrocarbon radical group consisting solely of carbon and hydrogen atoms, containing at least one triple bond, having from two to ten carbon atoms (i.e., C ₂ -C ₁₀ alkynyl). In all cases, when it appears in this application, a range of numerical values such as "2-10" refers to each integer in a given range; for example, “2-10 carbon atoms” means that an alkynyl group may consist of 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms. In some embodiments, alkynyl contains from two to eight carbon atoms. In other embodiments, alkynyl has from two to five carbon atoms (e.g., C ₂ -C ₅ alkynyl). Alkynyl is attached to the remainder of the molecule via a single bond, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl and the like. Unless specifically indicated otherwise in this description, the alkynyl group is optionally substituted with one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl , trifluoromethoxy, nitro, trimethylsilanyl, -OR ^a , -SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -OC (O) N (R ^a ) ₂ , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N (R ^a ) C (O) N (R ^a ) ₂ , N (R ^a ) C (NR ^a ) N (R ^a ) ₂ , -N (R ^a ) S (O) _t R ^a (where t is 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a ) ₂ , where each R ^a independently represents hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Alkynyl-cycloalkyl” refers to a - (alkynyl) cycloalkyl radical, wherein alkynyl and cycloalkyl are as described herein and which are optionally substituted with one or more of one of the substituents described as suitable substituents for alkynyl and cycloalkyl, respectively .

"Carboxaldehyde" refers to the radical - (C = O) H.

"Carboxyl" refers to the radical - (C = O) OH.

“Cyano” refers to the —CN radical.

“Cycloalkyl” refers to a monocyclic or polycyclic radical that contains only carbon and hydrogen and may be saturated or partially unsaturated. Cycloalkyl groups include groups having 3-10 ring atoms (i.e., C ₃ -C ₁₀ cycloalkyl). In all cases, when it appears in this application, a range of numerical values such as "3-10" refers to each integer in a given range; for example, “3-10 carbon atoms” means that a cycloalkyl group may consist of 3 carbon atoms, etc., up to and including 10 carbon atoms. In some embodiments, it is a C ₃ -C ₈ cycloalkyl radical. In some embodiments, it is a C ₃ -C ₅ cycloalkyl radical. Illustrative examples of cycloalkyl groups include, but are not limited to, the following groups: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl and the like. Unless specifically indicated otherwise in this description, the cycloalkyl group is optionally substituted with one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl , trifluoromethoxy, nitro, trimethylsilanyl, -OR ^a , -SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -OC (O) N (R ^a ) ₂ , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N (R ^a ) C (O) N (R ^a ) ₂ , N (R ^a ) C (NR ^a ) N (R ^a ) ₂ , -N (R ^a ) S (O) _t R ^a (where t is 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a ) ₂ , where each R ^a independently represents hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Cycloalkyl-alkenyl” refers to a - (cycloalkyl) alkenyl radical, wherein cycloalkyl and heterocycloalkyl are as described herein and which are optionally substituted with one or more of one of the substituents described as suitable substituents for heterocycloalkyl and cycloalkyl, respectively .

“Cycloalkyl-heterocycloalkyl” refers to a - (cycloalkyl) heterocyclyl radical, wherein cycloalkyl and heterocycloalkyl are as described herein and which are optionally substituted with one or more of the substituents described as suitable substituents for heterocycloalkyl and cycloalkyl, respectively.

“Cycloalkyl-heteroaryl” refers to a - (cycloalkyl) heteroaryl radical, wherein cycloalkyl and heterocycloalkyl are as described herein and which are optionally substituted with one or more of one of the substituents described as suitable substituents for heterocycloalkyl and cycloalkyl, respectively .

The term “alkoxy” refers to an —O-alkyl group comprising 1-8 carbon atoms in a straight, branched, cyclic configuration, and combinations thereof, attached to the parent structure through an oxygen atom. Examples include methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclohexyloxy and the like. "Lower alkoxy" refers to alkoxy groups containing from one to six carbon atoms. In some embodiments, C ₁ -C ₄ alkyl is an alkyl group that encompasses straight and branched chain alkyls of 1-4 carbon atoms.

The term "substituted alkoxy" refers to an alkoxy group wherein the alkyl moiety is substituted (i.e., -O- (substituted alkyl)). Unless specifically indicated otherwise in this description, the alkyl group of the alkoxy group is optionally substituted with one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR ^a , SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -OC (O) N (R ^a ) ₂ , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N (R ^a ) C (O) N (R ^a ) ₂ , N (R ^a ) C (NR ^a ) N (R ^a ) ₂ , -N (R ^a ) S (O), R ^a (where t is 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a ) ₂ , where each R ^a independently represents hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

The term “alkoxycarbonyl” refers to a group of the formula (alkoxy) (C = O) - attached via carbonyl carbon, wherein the alkoxy group has the indicated number of carbon atoms. Thus, a C ₁ -C ₆ alkoxycarbonyl group is an alkoxy group having 1-6 carbon atoms attached via their oxygen to the carbonyl linker. “Lower alkoxycarbonyl" refers to an alkoxycarbonyl group, wherein the alkoxy group is a lower alkoxy group. In some embodiments, C ₁ -C ₄ alkoxy is an alkoxy group which encompasses straight and branched chain alkoxy groups of 1-4 carbon atoms.

The term “substituted alkoxycarbonyl” refers to a group (substituted alkyl) —O — C (O) -, where this group is attached to the parent structure through a carbonyl functional group. Unless specifically indicated otherwise in this description, the alkyl group of the alkoxycarbonyl group is optionally substituted with one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR ^a , SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -OC (O) N (R ^a ) ₂ , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N (R ^a ) C (O) N (R ^a ) ₂ , N (R ^a ) C (NR ^a ) N (R ^a ) ₂ , -N (R ^a ) S (O), R ^a (where t is 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ ( where t is 1 or 2) or PO ₃ (R ^a ) ₂ , where each R ^a independently represents hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

"Acyl" refers to the groups (alkyl) -C (O) -, (aryl) -C (O) -, (heteroaryl) -C (O) -, (heteroalkyl) -C (O) - and (heterocycloalkyl) - C (O) -, where this group is attached to the original structure through a carbonyl functional group. In some embodiments, it is a C ₁ -C ₁₀ acyl radical, which means the total number of atoms in the chain or ring of the alkyl, aryl, heteroaryl or heterocycloalkyl part of the acyloxy group plus the acyl carbonyl carbon, i.e. the other three atoms in the ring or chain plus carbonyl. If the radical R is heteroaryl or heterocycloalkyl, then heteroatoms in the ring or atoms in the chain contribute to the total number of atoms in the chain or in the ring. Unless specifically indicated otherwise in this description, the “R” acyloxy group is optionally substituted with one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano , trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR ^a , SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , - OC (O) N (R ^a ) ₂ , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , - N (R ^a ) C (O) N (R ^a ) ₂ , N (R ^a ) C (NR ^a ) N (R ^a ) ₂ , -N (R ^a ) S (O), R ^a (where t is 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a ) ₂ , g e each R ^a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Acyloxy” refers to the radical R (C = O) O—, where “R” is alkyl, aryl, heteroaryl, heteroalkyl or heterocycloalkyl, as described herein. In some embodiments, it is a C ₁ -C ₄ acyloxy radical, which means the total number of atoms in the chain or in the ring of the alkyl, aryl, heteroaryl or heterocycloalkyl part of the acyloxy group plus carbon of the acyl carbonyl group, i.e. the other three atoms in the ring or chain plus carbonyl. If the radical R is heteroaryl or heterocycloalkyl, then heteroatoms in the ring or atoms in the chain contribute to the total number of atoms in the chain or in the ring. Unless specifically indicated otherwise in this description, the “R” acyloxy group is optionally substituted with one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano , trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR ^a , -SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -OC (O) N (R ^a ) ₂ , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N (R ^a ) C (O) N (R ^a ) ₂ , N (R ^a ) C (NR ^a ) N (R ^a ) ₂ , -N (R ^a ) S (O) _t R ^a (where t 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a ) ₂ where each R ^a independently represents hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Amino” or “amine” refers to the radical group —N (R ^a ) ₂ , where each R ^a independently represents hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl, if not specifically stated otherwise in this description. If the group —N (R ^a ) ₂ has two non-hydrogen R ^a , then they can be combined with a nitrogen atom to form a 4-, 5-, 6- or 7-membered ring. For example, it is understood that —N (R ^a ) ₂ includes, but is not limited to, 1-pyrrolidinyl and 4-morpholinyl. Unless specifically indicated otherwise in this description, the amino group is optionally substituted with one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR ^a , -SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -OC ( O) N (R ^a ) ₂ , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N ( R ^a ) C (O) N (R ^a ) ₂ , N (R ^a ) C (NR ^a ) N (R ^a ) ₂ , -N (R ^a ) S (O), R ^a (where t is 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a ) ₂ , where to zhdy R ^a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl, and each of these groups may optionally be substituted as defined herein.

The term “substituted amino” also refers to the N-oxides of each of the —NHR ^d and NR ^d R ^d groups as described above. N-oxides can be obtained by treating the corresponding amino group, for example, hydrogen peroxide or m-chloroperoxybenzoic acid. The person skilled in the art knows the reaction conditions for conducting N-oxidation.

“Amide” or “amido” refers to a chemical moiety having the formula —C (O) N (R) ₂ or —NHC (O) R, wherein R is selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl ( bonded via ring carbon) and a heteroalicyclic radical (bonded via ring carbon), and each group itself may possibly be substituted. In some embodiments, it is a C ₁ -C ₄ amido radical or an amide radical, and the carbonyl amide carbon atom is included in the total number of carbon atoms in this radical. R ² in the —N (R) ₂ amide may optionally be taken together with the nitrogen to which it is attached to form a 4-, 5-, 6- or 7-membered ring. Unless specifically indicated otherwise in this description, the amido group is optionally independently substituted with one or more of the substituents described in this application for alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl. The amide may be an amino acid or peptide molecule attached to a compound of formula (I), resulting in a prodrug. Any amine, hydroxy or carboxyl-containing side chain in the compounds set forth herein may be amidated. Techniques and specific groups for preparing such amides are known to those skilled in the art and can be easily found in reference materials such as Greene and Wuts, Protective Groups in Organic Synthesis, 3rd ed., John Wiley & Sons, New York, NY, 1999, which is incorporated herein by reference in its entirety.

The term “aromatic” or “aryl” refers to an aromatic radical containing from six to ten ring atoms (for example, a C ₆ -C ₁₀ aromatic radical or a C ₆ -C ₁₀ aryl) that has at least one ring containing a conjugated system pi-electrons, which is carbocyclic (e.g. phenyl, fluorenyl and naphthyl). Bivalent radicals formed from substituted benzene derivatives and having free valencies on ring atoms are called substituted phenylene radicals. Bivalent radicals formed from monovalent polycyclic hydrocarbon radicals, the names of which end with "-yl", as a result of the removal of one hydrogen atom from a carbon atom with a free valency, are called by adding an "-iden" to the name of the corresponding monovalent radical, for example, a naphthyl group with two points of attachment is called naphthylidene. In all cases, when it appears in this application, a range of numerical values such as "6-10" refers to each integer in a given range; for example, “6-10 ring atoms” means that an aryl group may consist of 6 ring atoms, 7 ring atoms, etc., up to and including 10 ring atoms. The term includes monocyclic or polycyclic formed from fused rings (i.e. rings that share common adjacent pairs of ring atoms), groups. Unless specifically indicated otherwise in this description, the aryl moiety is optionally substituted with one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl , trifluoromethoxy, nitro, trimethylsilanyl, -OR ^a , -SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -OC (O) N (R ^a ) ₂ , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N (R ^a ) C (O) N (R ^a ) ₂ , N (R ^a ) C (NR ^a ) N (R ^a ) ₂ , -N (R ^a ) S (O), R ^a (where t is 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a ) ₂ , where each R ^a independently represents hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Aralkyl” or “arylalkyl” refers to an (aryl) alkyl radical, wherein aryl and alkyl are as set forth herein and which are optionally substituted with one or more of one of the substituents described as suitable substituents for aryl and alkyl, respectively.

“Ester” refers to a chemical radical of the formula —COOR, where R is selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded via ring carbon) and a heteroalicyclic radical (bonded through ring carbon). Any amine, hydroxy, or carboxyl-containing side chain in the compounds described herein may be esterified. Techniques and specific groups for preparing such esters are known to those skilled in the art and can be readily found in reference materials such as Greene and Wuts, Protective Groups in Organic Synthesis, 3rd ed., John Wiley & Sons, New York, NY, 1999, which is incorporated herein by reference in its entirety. Unless specifically indicated otherwise in this description, the ester group is optionally substituted with one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl , trifluoromethoxy, nitro, trimethylsilanyl, -OR ^a , -SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -OC (O) N (R ^a ) ₂ , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N (R ^a ) C (O) N (R ^a ) ₂ , N (R ^a ) C (NR ^a ) N (R ^a ) ₂ , -N (R ^a ) S (O) _t R ^a (where t is 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a ) ₂ , where each R ^a independently represents hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Fluoroalkyl” refers to an alkyl radical as defined above that is substituted with one or more fluoro radicals as defined above, for example trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl and the like. The alkyl portion of the fluoroalkyl radical may optionally be substituted as described above for the alkyl group.

"Halo", "halogen" or, alternatively, "halogen" means fluoro, chloro, bromo or iodo. The terms “haloalkyl”, “haloalkenyl”, “haloalkynyl” and “haloalkoxy” include alkyl, alkenyl, alkynyl and alkoxy structures substituted with one or more halogen groups or combinations thereof. For example, the terms “fluoroalkyl” and “fluoroalkoxy” include haloalkyl and haloalkoxy groups, respectively, in which halogen is fluoro.

“Heteroalkyl”, “heteroalkenyl” and “heteroalkynyl” include optionally substituted alkyl, alkenyl and alkynyl radicals that have one or more skeletal atoms in a chain selected from a non-carbon atom, for example oxygen, nitrogen, sulfur, phosphorus, or combinations thereof . A range of numerical values may be given, for example C ₁ -C ₄ heteroalkyl, referring to the length of the chain as a whole, the length of which in this example is 4 atoms. For example, the —CH ₂ OCH ₂ CH ₃ radical is referred to as “C ₄ ” heteroalkyl, which contains a heteroatom center in a atomic description of the chain length. Attachment to the remainder of the molecule can be carried out either through a heteroatom or carbon in a heteroalkyl chain. The heteroalkyl group may be substituted by one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo, trimethylsilany ^a , -SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N (R ^a ) S (O), R ^a (where t is 1 or 2), - S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a ) ₂ , where each R ^{a is} independently represents hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalki l, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Heteroalkylaryl” refers to a - (heteroalkyl) aryl radical, wherein heteroalkyl and aryl are as described herein and which are optionally substituted with one or more of one of the substituents described as suitable substituents for heteroalkyl and aryl, respectively.

“Heteroalkylheteroaryl” refers to a - (heteroalkyl) heteroaryl radical, wherein heteroalkyl and heteroaryl are as described herein and which are optionally substituted with one or more of one of the substituents described as suitable substituents for heteroalkyl and heteroaryl, respectively.

“Heteroalkylheterocycloalkyl” refers to a - (heteroalkyl) heterocycloalkyl radical, wherein heteroalkyl and heteroaryl are as described herein and which are optionally substituted with one or more of the substituents described as suitable substituents for heteroalkyl and heterocycloalkyl, respectively .

“Heteroalkylcycloalkyl” refers to a - (heteroalkyl) cycloalkyl radical, wherein heteroalkyl and cycloalkyl are as described herein and which are optionally substituted with one or more of the substituents described as suitable substituents for heteroalkyl and cycloalkyl, respectively.

“Heteroaryl” or, alternatively, “heteroaromatic” refers to a 5-18 membered aromatic radical (eg, C ₅ -C ₁₃ heteroaryl) that includes one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur, and which may be monocyclic, bicyclic, tricyclic or tetracyclic ring system. In all cases, when it appears in this application, a range of numerical values, such as "5-18", refers to each integer in a given range; for example, “5-18 ring atoms” means that a heteroaryl group may consist of 5 ring atoms, 6 ring atoms, etc., up to and including 18 ring atoms. Bivalent radicals formed from monovalent heteroaryl radicals whose names end in "-yl" by removing one hydrogen atom from a free valency atom are called by adding "-ideas" to the name of the corresponding monovalent radical, for example, a pyridyl group with two points addition is pyridylidene. An N-containing “heteroaromatic” or “heteroaryl” moiety refers to an aromatic group in which at least one of the skeletal ring atoms is a nitrogen atom. The polycyclic heteroaryl group may be fused or non fused. The heteroatom (s) in the heteroaryl radical are possibly oxidized (s). One or more nitrogen atoms, if present, are possibly quaternized. Heteroaryl is attached to the remainder of the molecule through any atom of the ring (ec). Examples of heteroaryl include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzooxazolyl, benzo [d] thiazolyl, benzothiadiazolyl, benzo [b] [1,4] dioxepinyl, benzo [b] [1,4] oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzoxazolyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzofurazanyl, benzothiazhenyl, 2-benzothiophenyl, 2-benzothiophenyl) benzotriazolyl, benzo [4,6] imidazo [1,2-a] pyridinyl, carbazoles , cinnolinyl, cyclopenta [d] pyrimidinyl, 6,7-dihydro-5H-cyclopenta [4,5] thieno [2,3-d] pyrimidinyl, 5,6-dihydrobenzo [h] quinazolinyl, 5,6-dihydrobenzo [h ] cinnolinyl, 6,7-dihydro-5H-benzo [6,7] cyclohepta [1,2-c] pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furazanyl, furanonyl, furo [3,2-c] pyridinyl, 5.6 7,8,9,10-hexahydrocyclo-octa [d] pyrimidinyl, 5,6,7,8,9,10-hexahydrocyclooct [d] pyridazinyl, 5,6,7,8,9,10-hexahydrocyclooct [d ] pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolisinyl, isoxazolyl, 5.8-methano -5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 5,6,6a, 7,8,9,10,10a-octahydrobenzo [h] quinazolinyl , 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrrolyl, pyrazolyl, pyrazolo [3,4-d] pyrimidinyl, pyridinyl, pyrido [3,2-d] pyrimidinyl, pyrido [3,4-d] pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo ] thieno [2,3-d] piri idinyl, 6,7,8,9-tetrahydro-5H-cyclohepta [4,5] thieno [2,3-d] pyrimidinyl, 5,6,7,8-tetrahydropyrido [4,5-c] pyridazinyl, thiazolyl, thiadiazolyl, thiopyranyl, triazolyl, tetrazolyl, triazinyl, thieno [2,3-d] pyrimidinyl, thieno [3,2-d] pyrimidinyl, thieno [2,3-c] pyridinyl and thiophenyl (i.e. thienyl). Unless specifically indicated otherwise in this description, the heteroaryl moiety is optionally substituted with one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, -OR ^a , -SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -C ( O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N (R ^a ) S (O) _t R ^a (where t is 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a ) ₂ , where each R ^a independently represents hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

Substituted heteroaryl also includes ring systems substituted with one or more oxide (-O-) substituents, such as pyridinyl-N-oxides.

“Heteroarylalkyl” refers to a moiety having an aryl moiety as described herein, coupled to an alkylene moiety as described herein, wherein such attachment to the remainder of the molecule is via an alkylene group.

“Heterocycloalkyl” refers to a stable 3-18 membered non-aromatic ring radical containing from two to twelve carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur. In all cases, when it appears in this application, a range of numerical values such as "3-18" refers to each integer in a given range; for example, “3-18 ring atoms” means that a heterocycloalkyl group may consist of 3 ring atoms, 4 ring atoms, etc., up to and including 18 ring atoms. In some embodiments, it is C ₅ -C ₁₀ heterocycloalkyl. In some embodiments, it is C ₄ -C ₁₀ heterocycloalkyl. In some embodiments, it is C ₃ -C ₁₀ heterocycloalkyl. Unless otherwise specifically indicated herein, a heterocycloalkyl radical is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems. The heteroatoms in the heterocycloalkyl radical may possibly be oxidized. One or more nitrogen atoms, if present, are possibly quaternized. The heterocycloalkyl radical is partially or fully saturated. Heterocycloalkyl can be attached to the remainder of the molecule through any atom of the ring (ec). Examples of such heterocycloalkyl radicals include, but are not limited to, dioxolanyl, thienyl [1,3] dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, oxohydrodinopyrhydro-2-dihydro-2-dihydrodinol , piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, tritianil, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl-1,1-di Inyl. Unless specifically indicated otherwise in this description, the heterocycloalkyl group is optionally substituted with one or more substituents independently representing alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro , oxo, thioxo, trimethylsilanyl, -OR ^a , -SR ^a , -OC (O) -R ^a , -N (R ^a ) ₂ , -C (O) R ^a , -C (O) OR ^a , -C (O) N (R ^a ) ₂ , -N (R ^a ) C (O) OR ^a , -N (R ^a ) C (O) R ^a , -N (R ^a ) S (O) _t R ^a ( where t is 1 or 2), -S (O) _t OR ^a (where t is 1 or 2), -S (O) _t N (R ^a ) ₂ (where t is 1 or 2) or PO ₃ (R ^a) ₂ where each R ^a is independently is hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heteroaryl or heteroarylalkyl.

"Heterocycloalkyl" also includes bicyclic ring systems, where one non-aromatic ring, usually consisting of 3-7 ring atoms, contains at least 2 carbon atoms in addition to 1-3 heteroatoms independently selected from oxygen, sulfur and nitrogen, as well as combinations containing at least one of the aforementioned heteroatoms; and the other ring, usually consisting of 3-7 ring atoms, possibly contains 1-3 heteroatoms independently selected from oxygen, sulfur and nitrogen, and is not aromatic.

"Isomers" are different compounds having the same molecular formula. "Stereoisomers" are isomers that differ only in the arrangement of these atoms in space. "Enantiomers" are a pair of stereoisomers that are non-overlapping mirror reflections of each other. A 1: 1 mixture of enantiomer pairs is a "racemic" mixture. The term "(±)" is used to indicate a racemic mixture, where appropriate. "Diastereoisomers" refers to stereoisomers having at least two asymmetric atoms, but which are not mirror images of each other. Absolute stereochemistry is determined by the Kahn-Ingold-Prelog R-S system. In the case where the compound is a pure enantiomer, the stereochemistry for each chiral carbon atom can be defined either as R or S. Separated compounds, the absolute configuration of which is unknown, can be designated as (+) or (-) depending on the direction (right- or levorotatory) in which they rotate plane-polarized light at the wavelength of the sodium D-line. Some of the compounds described in this application contain one or more asymmetric centers and therefore can be a source of enantiomers, diastereomers and other stereoisomeric forms, which can be defined in terms of absolute stereochemistry as (R) -or (S) -. It is implied that the proposed chemical structures, pharmaceutical compositions and methods include all such possible isomers, including racemic mixtures, optically pure forms and intermediate mixtures. Optically active (R) and (S) isomers can be prepared using chiral synthons or chiral reagents, or separated using conventional techniques. If the compounds described in this application contain olefinic double bonds or other centers of geometric asymmetry, and unless otherwise indicated, it is understood that the compounds include both E and Z geometric isomers.

"Grouping" refers to a particular segment or functional group of a molecule. Chemical groups are often understood to mean chemical structures embedded in or attached to a molecule.

“Nitro” refers to the radical —NO ₂ .

“Oxa” refers to the radical —O—.

"Oxo" refers to the radical = O.

"Tautomers" are structurally different isomers that interconvert as a result of tautomerization. "Tautomerization" is a form of isomerization and includes prototropic tautomerization or proton transfer tautomerization, which is considered a subgroup of the chemistry of acid-base interactions. It is understood that "prototropic tautomerization" or "proton transfer tautomerization" includes proton migration, which is accompanied by changes in the coupling ratio, often with the interconversion of a single bond and an adjacent double bond. If there is the possibility of tautomerization (for example, in solution), then the chemical equilibrium of the tautomers can be achieved. An example of tautomerization is keto-enol tautomerization. A specific example of keto-enol tautomerization is the interconversion of the tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one. Another example of tautomerization is phenol-keto-tautomerization. A specific example of phenol-keto-tautomerization is the interconversion of the tautomers of pyridin-4-ol and pyridin-4 (1H) -one.

Compounds of the present invention may also contain isotopes of atoms for one or more of the atoms that make up such compounds in unnatural proportions. For example, compounds can be radioactively labeled with radioactive isotopes, such as, for example, tritium ( ³ N), iodine-125 ( ¹²⁵ I) or carbon-14 ( ¹⁴ C). All isotopic variations of the compounds of the present invention, whether radioactive or not, fall within the scope of the present invention.

A “leaving group or atom” is any group or any atom which, under the reaction conditions, will be cleaved from the starting material, thereby facilitating the passage of the reaction to a particular site. Suitable examples of such groups, unless otherwise indicated, are halogen atoms, mesyloxy, p-nitrobenzenesulfonyloxy and tosyloxy groups.

The term “protecting group” has the meaning with which it is traditionally associated in organic synthesis, that is, it is a group that selectively blocks one or more reactive sites in a multifunctional compound, such that a chemical reaction can proceed selectively at another unprotected reactive site, and so that this group can be easily removed after the completion of the selective reaction. A number of protective groups are described, for example, in T.N. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, third edition, John Wiley & Sons, New York (1999). For example, a hydroxy-protected form is one wherein at least one of the hydroxy groups present in the compound is protected by a hydroxy-protecting group. Similarly, amines and other reactive groups can be protected.

"Solvate" refers to a compound (for example, a compound selected from a compound of formula I or a pharmaceutically acceptable salt thereof) in physical association with one or more pharmaceutically acceptable solvent molecules. Obviously, a “compound of formula I” embraces a compound of formula I and solvates of this compound, as well as mixtures thereof.

"Substituted" means that the group may be substituted with one or more additional groups (s), individually and independently selected from acyl, alkyl, alkylaryl, cycloalkyl, aralkyl, aryl, carbohydrate, carbonate, heteroaryl, heterocycloalkyl, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halogen, carbonyl, ester, thiocarbonyl, isocyanato, thiocyanato, isothiocyanato, nitro, oxo, perhaloalkyl, perfluoroalkyl, phosphate, silyl, sulfinyl, sulfonyl sulfonyl, phonate, urea and amino, including mono- and di-substituted amino groups and their protected derivatives. Di-substituted amino groups encompass those groups which form a ring together with the nitrogen of the amino group, such as, for example, morpholino. The substituents themselves may be substituted, for example, the cycloalkyl substituent may have a halide substituted at one or more than one carbon atom in the ring, and the like. The protecting groups that the protected derivatives of the above substituents can form are known to those skilled in the art and can be found in references such as Greene and Wuts mentioned above.

"Sulfanyl" refers to the groups: -S- (possibly substituted alkyl), -S- (possibly substituted aryl), -S- (possibly substituted heteroaryl) and -S- (possibly substituted heterocycloalkyl).

"Sulfinyl" refers to the groups: -S (O) -H, -S (O) - (possibly substituted alkyl), -S (O) - (possibly substituted amino), -S (O) - (possibly substituted aryl) , -S (O) - (possibly substituted heteroaryl) and -S (O) - (possibly substituted heterocycloalkyl).

“Sulfonyl” refers to the groups: —S (O ₂ ) —H, —S (O ₂ ) - (optionally substituted alkyl), —S (O ₂ ) - (optionally substituted amino), —B (O ₂ ) - ( optionally substituted aryl), —S (O ₂ ) - (optionally substituted heteroaryl) and —S (O ₂ ) - (optionally substituted heterocycloalkyl).

“Sulfonamidyl” or “sulfonamido” refers to the radical —S (= O) ₂ —NRR, where each R is independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl (bonded via ring carbon) and a heteroalicyclic radical (bonded through ring carbon). The R groups on the —NRR of the —S (= O) ₂ —NRR radical can be taken together with the nitrogen to which they are attached to form a 4-, 5-, 6- or 7-membered ring. In some embodiments, it is C ₁ -C ₁₀ sulfonamido, where each R in sulfonamido contains a total of 1 carbon, 2 carbon, 3 carbon, or 4 carbon. The sulfonamido group is optionally substituted with one or more of the substituents described for alkyl, cycloalkyl, aryl, heteroaryl, respectively.

“Sulfoxyl” refers to the radical —S (= O) ₂ OH.

“Sulfonate” refers to the radical —S (= O) ₂ —OR, where R is selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded via ring carbon) and a heteroalicyclic radical (bonded via ring carbon). The sulfonate group is optionally substituted at R with one or more of the substituents described for alkyl, cycloalkyl, aryl, heteroaryl, respectively.

If substituent groups are defined by their traditional chemical formulas, written from left to right, then they equally encompass chemically identical substituents that would result from writing the structure from right to left, for example, -CH ₂ O- is the equivalent of -OSH ₂ -.

The compounds of the present invention also include crystalline and amorphous forms of such compounds, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs and amorphous forms of compounds, as well as mixtures thereof. The "crystalline form", "polymorph" and "new form" can be used interchangeably in this application, and it is understood that they include all crystalline and amorphous forms of the compound, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs ( including anhydrates), conformational polymorphs and amorphous forms, as well as mixtures thereof, unless a particular crystalline or amorphous form is mentioned.

Chemical structures include, but are not limited to, compounds of formula I and all pharmaceutically acceptable forms thereof. Pharmaceutically acceptable forms of the compounds provided herein include pharmaceutically acceptable salts, chelates, non-covalent complexes, prodrugs, and mixtures thereof. In some embodiments, the compounds described herein are in the form of pharmaceutically acceptable salts. Thus, the terms “chemical structure” and “chemical structures” also encompass pharmaceutically acceptable salts, chelates, non-covalent complexes, prodrugs, and mixtures.

In addition, if the compound of formula I is obtained as an acid addition salt, the free base can be obtained by alkalizing the acid salt solution. Conversely, if the product is a free base, then the addition salt, in particular a pharmaceutically acceptable addition salt, can be obtained by dissolving the free base in a suitable organic solvent and treating the solution with an acid in accordance with traditional methods for preparing acid addition salts from basic compounds. Those skilled in the art will recognize various synthesis methodologies that can be used to produce non-toxic pharmaceutically acceptable addition salts.

In one aspect of the present invention, there is provided a compound of formula I.

or its pharmaceutically acceptable salt, where

W _d represents heterocycloalkyl, aryl or heteroaryl;

B represents an alkyl, amino, heteroalkyl or moiety of formula II:

,

,

where W _c represents aryl, heteroaryl, heterocycloalkyl or cycloalkyl, and

q is an integer of 0, 1, 2, 3 or 4;

X is absent or represents - (CH (R ⁹ )) _z -; and z is independently an integer of 1, 2, 3, or 4;

Y is absent, represents -O-, -S-, -S (= O) -, -S (= O) ₂ -, -N (R ⁹ ) -, -C (= O) - (CHR ⁹ ) _z -, -C (= O) -, -N (R ⁹ ) -C (= O) - or -N (R ⁹ ) -C (= O) NH-, -N (R ⁹ ) C (R ⁹ ) ₂ - or -C (= O) - (CHR ⁹ ) _z -;

R ¹ represents hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, hydroxy urea or carbonate;

R ² represents alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro, hydroxy, nitro, nitro, hydroxy, nitro carbonate;

R ³ represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro, aryl or heteroaryl;

R ⁵ , R ⁶ , R ⁷ and R ⁸ independently represent hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₅ alkenyl, C ₂ -C ₅ alkynyl, C ₃ -C ₅ cycloalkyl, C ₁ -C ₄ heteroalkyl , C ₁ -C ₄ alkoxy, C ₁ -C ₄ amido, amino, acyl, C ₁ -C ₄ acyloxy, C ₁ -C ₄ sulfonamido, halo, cyano, hydroxy or nitro; and

in each case, R ⁹ independently represents hydrogen, d-Syalkyl, C ₃ -C ₇ cycloalkyl, heterocycloalkyl or C ₂ -C ₁₀ heteroalkyl.

In some embodiments, B is unsubstituted or substituted alkyl, including, but not limited to, - (CH ₂ ) ₂ —NR ^a R ^a , where each R ^{a is} independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl , heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl, or NR ^a R ^{a are} joined together to form a cyclic moiety which includes, but is not limited to, piperidinyl, piperazinyl and morpholinyl. In some embodiments, B is unsubstituted or substituted amino. In some embodiments, B is unsubstituted or substituted heteroalkyl.

.

.

In some embodiments, B is a grouping of formula II, where W _c is a member selected from the group consisting of unsubstituted or substituted aryl, substituted phenyl, unsubstituted or substituted heteroaryl, including, but not limited to, pyridin-2-yl, pyridine -3-yl, pyridin-4-yl, pyrimidin-4-yl, pyrimidin-2-yl, pyrimidin-5-yl or pyrazin-2-yl, unsubstituted or substituted monocyclic heteroaryl, unsubstituted or substituted bicyclic heteroaryl, heteroaryl containing two heteroatoms as co atom atoms, unsubstituted or substituted heteroaryl containing a ring nitrogen atom, heteroaryl containing two ring nitrogen atoms, heteroaryl containing nitrogen and sulfur as ring atoms, unsubstituted or substituted heterocycloalkyl, including, but not limited to, morpholinyl, tetrahydropyranyl, and piper piperidinyl, unsubstituted or substituted cycloalkyl, including, but not limited to cyclopentyl and cyclohexyl.

In some embodiments, B represents one of the following groups:

-CH ₃ -CH ₂ CH ₃ -CH (CH ₃ ) ₂

.

.

In some embodiments, B is substituted by one or more than one of alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy or nitro, when each alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amido, amino, acyl, acyloxy or sulfonamido itself may be substituted.

In some embodiments, R ¹ is a member selected from the group consisting of hydrogen, unsubstituted or substituted alkyl, unsubstituted or substituted heteroalkyl, unsubstituted or substituted alkenyl, unsubstituted or substituted alkynyl, unsubstituted or substituted cycloalkyl, or unsubstituted or substituted heterocycloalkyl. In some embodiments, R ¹ is unsubstituted or substituted aryl, unsubstituted or substituted arylalkyl, unsubstituted or substituted heteroaryl, or unsubstituted or substituted heteroarylalkyl. In some embodiments, R ¹ is unsubstituted or substituted alkoxy, unsubstituted or substituted amido, unsubstituted or substituted amino. In some embodiments, R ¹ is unsubstituted or substituted acyl, unsubstituted or substituted acyloxy, unsubstituted or substituted alkoxycarbonyl, or unsubstituted or substituted sulfonamido. In some embodiments, R ¹ is halogen, which includes —Cl, —F, —I, and —Br. In some embodiments, R ^{1 is} selected from the group consisting of cyano, hydroxy, nitro, unsubstituted or substituted phosphate, unsubstituted or substituted urea and carbonate.

In some embodiments, when R ¹ is alkyl, R ¹ is methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, w-butyl, pentyl, hexyl or heptyl.

In some embodiments, when R ¹ is alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, or hydroxy, R ¹ is substituted with phosphate, or unsubstituted urea, or substituted urea, or carboxylic acid or carbonate.

In some embodiments, when R ¹ is alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl or sulfonamido, more than one is substituted by R ¹ one of alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy nitro or , but kenil, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, or sulfonamido may itself be substituted.

In some embodiments, R ² is a member selected from the group consisting of unsubstituted or substituted alkyl, unsubstituted or substituted heteroalkyl, unsubstituted or substituted alkenyl, unsubstituted or substituted alkynyl, unsubstituted or substituted cycloalkyl, and unsubstituted or substituted heterocycloalkyl. In some embodiments, R ² is unsubstituted or substituted aryl, unsubstituted or substituted arylalkyl, unsubstituted or substituted heteroaryl, or unsubstituted or substituted heteroarylalkyl. In some embodiments, R ² is unsubstituted or substituted alkoxy, unsubstituted or substituted amido, unsubstituted or substituted amino. In some embodiments, R ² is unsubstituted or substituted acyl, unsubstituted or substituted acyloxy, unsubstituted or substituted alkoxycarbonyl, or unsubstituted or substituted sulfonamido. In some embodiments, R ² is halogen, which is —I, —F, —Cl, or —Br. In some embodiments, R ^{2 is} selected from the group consisting of cyano, hydroxy, nitro, carboxylic acid, and carbonate. In some embodiments, R ² is unsubstituted or substituted phosphate. In some embodiments, R ² is unsubstituted or substituted urea. In some embodiments, when R ² is alkyl, R ² is methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, sec-butyl, pentyl, hexyl or heptyl.

In some embodiments, when R ² is alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido or hydroxy, it is substituted urea or substituted by carbonate.

In some embodiments, when R ² is alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl or sulfonamido, it is substituted by more than one or from alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy or nitro, with each alkyl, heteroalkyl, alkenyl cycloalkyl terocycloalkyl, aryl, heteroaryl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl or sulfonamido may itself be substituted.

In some embodiments, q is an integer 0. In some embodiments, q is an integer 1. In some embodiments, q is an integer 2. In some embodiments, q is an integer 3. In some embodiments, q is an integer 4.

In some embodiments of a compound of formula I, IR ³ is a member selected from the group consisting of hydrogen, unsubstituted or substituted alkyl, unsubstituted or substituted alkenyl, and unsubstituted or substituted alkynyl. In some embodiments, R ³ is unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, unsubstituted or substituted cycloalkyl, or unsubstituted or substituted heterocycloalkyl. In some embodiments, R ³ is unsubstituted or substituted alkoxy, unsubstituted or substituted amido, unsubstituted or substituted amino. In some embodiments, R ³ is unsubstituted or substituted acyl, unsubstituted or substituted acyloxy, unsubstituted or substituted alkoxycarbonyl, or unsubstituted or substituted sulfonamido. In some embodiments, R ³ is halogen, which is —I, —F, —Cl, or —Br.

In some embodiments, R ^{3 is} selected from the group consisting of cyano, hydroxy and nitro. In some embodiments, when R ³ is alkyl, R ³ is methyl, ethyl, propyl, isopropyl, w-butyl, tert-butyl, sec-butyl, pentyl, hexyl or heptyl. In some embodiments, R ³ is —CF ₃ .

In some embodiments, when R ³ is alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl or sulfonamido, it is substituted with one or more of alkyl, heteroalkyl, alkenyl , alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy or nitro, each alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclo alkoxy, Medo, amino, acyl, acyloxy, alkoxycarbonyl, or sulfonamido may itself be substituted.

In some embodiments of a compound of formula I, IR ⁵ is hydrogen, unsubstituted or substituted alkyl (including, but not limited to, unsubstituted or substituted C ₁ -C ₄ alkyl). In some embodiments, R ⁵ is unsubstituted or substituted alkenyl, including, but not limited to, unsubstituted or substituted C ₂ -C ₆ alkenyl. In some embodiments, R ⁵ is unsubstituted or substituted alkynyl, including, but not limited to, unsubstituted or substituted C ₂ -C ₆ alkynyl. In some embodiments, R ⁵ is unsubstituted or substituted cycloalkyl, including, but not limited to, unsubstituted or substituted C ₃ -C ₅ cycloalkyl. In some embodiments, R ⁵ is unsubstituted or substituted heterocycloalkyl. In some embodiments, R ⁵ is unsubstituted or substituted heteroalkyl, including, but not limited to, unsubstituted or substituted C ₁ -C ₄ heteroalkyl. In some embodiments, R ⁵ is unsubstituted or substituted alkoxy, including, but not limited to, unsubstituted or substituted C ₁ -C ₄ alkoxy. In some embodiments, R ⁵ is an unsubstituted or substituted amido, including, but not limited to, unsubstituted or substituted C ₁ -C ₄ amido. In some embodiments, R ⁵ is unsubstituted or substituted amino. In some embodiments, R ⁵ is unsubstituted or substituted acyl, unsubstituted or substituted acyloxy, unsubstituted or substituted C ₁ -C ₄ acyloxy, unsubstituted or substituted alkoxycarbonyl, unsubstituted or substituted sulfonamido or unsubstituted or substituted C ₁ -C ₄ sulfonamido. In some embodiments, R ⁵ is halogen, which is —I, —F, —Cl, or —Br. In some embodiments, R ^{5 is} selected from the group consisting of cyano, hydroxy and nitro. In some other embodiments, R ⁵ is —CH ₃ , —CH ₂ CH ₃ , n-propyl, isopropyl, —OCH ₃ , —OCH ₂ CH _3, or —CF ₃ .

In some embodiments, when R ⁵ is alkyl, alkenyl, alkynyl, cycloalkyl, heteroalkyl, acyl, alkoxy, amido, amino, acyloxy, alkoxycarbonyl or sulfonamido, R ^{5 is} optionally substituted with one or more of alkyl, heteroalkyl, alkenyl, alkynyl , cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy or nitro, each alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, heterocycloalkyl, , amido, amino, and yl, acyloxy, alkoxycarbonyl, or sulfonamido may itself be substituted.

In some embodiments of a compound of formula I, IR ⁶ is hydrogen, unsubstituted or substituted alkyl (including, but not limited to, unsubstituted or substituted C ₁ -C ₄ alkyl). In some embodiments, R ⁶ is unsubstituted or substituted alkenyl, including, but not limited to, unsubstituted or substituted C ₂ -C ₅ alkenyl. In some embodiments, R ⁶ is unsubstituted or substituted alkynyl, including, but not limited to, unsubstituted or substituted C ₂ -C ₅ alkynyl. In some embodiments, R ⁶ is unsubstituted or substituted cycloalkyl, including, but not limited to, unsubstituted or substituted C ₃ -C ₅ cycloalkyl. In some embodiments, R ⁶ is unsubstituted or substituted heterocycloalkyl. In some embodiments, R ⁶ is unsubstituted or substituted heteroalkyl, including, but not limited to, unsubstituted or substituted C ₁ -C ₄ heteroalkyl. In some embodiments, R ⁶ is unsubstituted or substituted alkoxy, including, but not limited to, unsubstituted or substituted C ₁ -C ₄ alkoxy. In some embodiments, R ⁶ is an unsubstituted or substituted amido, including, but not limited to, unsubstituted or substituted C ₁ -C ₄ amido. In some embodiments, R ⁶ is unsubstituted or substituted amino. In some embodiments, R ⁶ is unsubstituted or substituted acyl, unsubstituted or substituted acyloxy, unsubstituted or substituted C ₁ -C ₄ acyloxy, unsubstituted or substituted alkoxycarbonyl, unsubstituted or substituted sulfonamido, or unsubstituted or substituted C ₁ -C ₄ sulfonamido. In some embodiments, R ⁶ is halogen, which is —I, —F, —Cl, or —Br. In some embodiments, R ^{6 is} selected from the group consisting of cyano, hydroxy and nitro. In some other embodiments, R ⁶ is —CH ₃ , —CH ₂ CH ₃ , n-propyl, isopropyl, —OCH ₃ , —OCH ₂ CH _3, or —CF ₃ .

In some embodiments, when R ⁶ is alkyl, alkenyl, alkynyl, cycloalkyl, heteroalkyl, acyl, alkoxy, amido, amino, acyloxy, alkoxycarbonyl or sulfonamido, R ^{6 is} optionally substituted with one or more of alkyl, heteroalkyl, alkenyl, alkynyl , cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy or nitro, each alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, heterocycloalkyl, , amido, amino, and yl, acyloxy, alkoxycarbonyl, or sulfonamido may itself be substituted.

In some embodiments of a compound of formula I, IR ⁷ is hydrogen, unsubstituted or substituted alkyl (including, but not limited to, unsubstituted or substituted C ₁ -C ₄ alkyl). In some embodiments, R ⁷ is unsubstituted or substituted alkenyl, including, but not limited to, unsubstituted or substituted C ₂ -C ₅ alkenyl. In some embodiments, R ⁷ is unsubstituted or substituted alkynyl, including, but not limited to, unsubstituted or substituted C ₂ -C ₅ alkynyl. In some embodiments, R ⁷ is unsubstituted or substituted cycloalkyl, including, but not limited to, unsubstituted or substituted C ₃ -C ₅ cycloalkyl. In some embodiments, R ⁷ is unsubstituted or substituted heterocycloalkyl. In some embodiments, R ⁷ is unsubstituted or substituted heteroalkyl, including, but not limited to, unsubstituted or substituted C ₁ -C ₄ heteroalkyl. in some embodiments, R ⁷ is unsubstituted or substituted alkoxy, including, but not limited to, unsubstituted or substituted C ₁ -C ₄ alkoxy. In some embodiments, R ⁷ is unsubstituted or substituted amido, including, but not limited to, unsubstituted or substituted C ₁ -C ₄ amido. In some embodiments, R ⁷ is unsubstituted or substituted amino. In some embodiments, R ⁷ is unsubstituted or substituted acyl, unsubstituted or substituted acyloxy, unsubstituted or substituted C ₁ -C ₄ acyloxy, unsubstituted or substituted alkoxycarbonyl, unsubstituted or substituted sulfonamido, or unsubstituted or substituted C ₁ -C ₄ sulfonamido. In some embodiments, R ⁷ is halogen, which is —I, —F, —Cl, or —Br. In some embodiments, R ^{7 is} selected from the group consisting of cyano, hydroxy and nitro. In some other embodiments, R ⁷ is —CH ₃ , —CH ₂ CH ₃ , n-propyl, isopropyl, —OCH ₃ , —OCH ₂ CH _3, or —CF ₃ .

In some embodiments, when R ⁷ is alkyl, alkenyl, alkynyl, cycloalkyl, heteroalkyl, acyl, alkoxy, amido, amino, acyloxy, alkoxycarbonyl or sulfonamido, R ^{7 is} optionally substituted with one or more of alkyl, heteroalkyl, alkenyl, alkynyl , cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy or nitro, each alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, heterocycloalkyl, , amido, amino, and yl, acyloxy, alkoxycarbonyl, or sulfonamido may itself be substituted.

In some embodiments of a compound of formula I, IR ⁸ is hydrogen, unsubstituted or substituted alkyl (including, but not limited to, unsubstituted or substituted C ₁ -C ₄ alkyl). In some embodiments, R ⁸ is unsubstituted or substituted alkenyl, including, but not limited to, unsubstituted or substituted C ₂ -C ₅ alkenyl. In some embodiments, R ⁸ is unsubstituted or substituted alkynyl, including, but not limited to, unsubstituted or substituted C ₂ -C ₅ alkynyl. In some embodiments, R ⁸ is unsubstituted or substituted cycloalkyl, including, but not limited to, unsubstituted or substituted C ₃ -C ₅ cycloalkyl. In some embodiments, R ⁸ is unsubstituted or substituted heterocycloalkyl. In some embodiments, R ⁸ is unsubstituted or substituted heteroalkyl, including, but not limited to, unsubstituted or substituted C ₁ -C ₄ heteroalkyl. In some embodiments, R ⁸ is unsubstituted or substituted alkoxy, including, but not limited to, unsubstituted or substituted C ₁ -C ₄ alkoxy. In some embodiments, R ⁸ is an unsubstituted or substituted amido, including, but not limited to, unsubstituted or substituted C ₁ -C ₄ amido. In some embodiments, R ⁸ is unsubstituted or substituted amino. In some embodiments, R ⁸ is unsubstituted or substituted acyl, unsubstituted or substituted acyloxy, unsubstituted or substituted C ₁ -C ₄ acyloxy, unsubstituted or substituted alkoxycarbonyl, unsubstituted or substituted sulfonamido, or unsubstituted or substituted C ₁ -C ₄ sulfonamido. In some embodiments, R ⁸ is halogen, which is —I, —F, —Cl, or —Br. In some embodiments, R ^{8 is} selected from the group consisting of cyano, hydroxy and nitro. In some other embodiments, R ⁸ is —CH ₃ , —CH ₂ CH ₃ , n-propyl, isopropyl, —OCH ₃ , —OCH ₂ CH _3, or —CF ₃ .

In some embodiments, when R ⁸ is alkyl, alkenyl, alkynyl, cycloalkyl, heteroalkyl, acyl, alkoxy, amido, amino, acyloxy, alkoxycarbonyl or sulfonamido, R ^{8 is} optionally substituted with one or more of alkyl, heteroalkyl, alkenyl, alkynyl , cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy or nitro, each alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, heterocycloalkyl, , amido, amino, and yl, acyloxy, alkoxycarbonyl, or sulfonamido may itself be substituted.

In some embodiments of the compounds of formula IR ⁵ , R ⁶ , R ⁷ and R ⁸ represent N.

In some embodiments, the compounds of formula IX are absent. In some embodiments, X is - (CH (R ⁹ )) _z ; and z is independently an integer of 1, 2, 3, or 4.

In some embodiments, R ⁹ is unsubstituted or substituted alkyl, including, but not limited to, unsubstituted or substituted C ₁ -C ₁₀ alkyl. In some embodiments, R ⁹ is unsubstituted or substituted cycloalkyl, including, but not limited to, unsubstituted or substituted C ₃ -C ₇ cycloalkyl. In some embodiments, R ⁹ is methyl or hydrogen. In some embodiments, R ⁹ is unsubstituted or substituted heterocycloalkyl, including, but not limited to, unsubstituted or substituted C ₂ -C ₁₀ heteroalkyl. In some embodiments, R ⁹ is unsubstituted or substituted heteroalkyl, including, but not limited to, unsubstituted or substituted C ₂ -C ₁₀ heteroalkyl.

When R ⁹ is any of the above, in some embodiments X is —CH ₂ -, —CH ₂ CH ₂ -, —CH ₂ CH ₂ CH ₂ -, —CH (CH ₃ ) —or —CH (CH ₂ CH ₃ ) -. In some embodiments, when X is —CH (CH ₃ ) -, —CH (CH ₃ ) - is in the (S) - or (R) stereochemical configuration.

In some embodiments, the compounds of formula IY are absent. In some embodiments, Y is —O—, —S—, —S (= 0) -, —S (= O) ₂ -, —C (= O) -, —N (R ⁹ ) (C = O) -, -N (R ⁹ ) (C = O) NH-, -N (R ⁹ ) C (R ⁹ ) ₂ - (such as, for example, -N (R ⁹ ) CH ₂ -, specifically -N (CH ₃ ) CH ₂ -, N (CH (CH ₃ ) ₂ ) CH ₂ - or N (CH ₂ CH ₃ ) CH ₂ -), -N (R ⁹ ) -, -N (CH ₃ ) -, -N (CH ₂ CH ₃ ) - or -N (CH (CH ₃ ) ₂ ) -. In some embodiments, Y represents —C (═O) - (CHR ⁹ ) z—; and z is independently an integer of 1, 2, 3, or 4.

In some embodiments, XY is —CH ₂ -, —CH ₂ —N (CH ₃ ), —CH (CH ₃ ) —NH—, (S) —CH (GH ₃ ) —NH—, or (R) —CH ( CH ₃ ) —NH—. In some embodiments, XY is —N (CH ₃ ) —CH ₂ -, N (CH ₂ CH ₃ ) CH ₂ -, —N (CH (CH ₃ ) ₂ ) CH ₂ -, or —NHCH ₂ -.

In some embodiments of the compounds of formula IW, _d is a member selected from the group consisting of unsubstituted or substituted heterocycloalkyl, unsubstituted or substituted aryl, and unsubstituted or substituted heteroaryl. In some embodiments, W _d is unsubstituted or substituted monocyclic heteroaryl or unsubstituted or substituted bicyclic heteroaryl. In some embodiments, W _d is a bicyclic heteroaryl having at least one heteroatom, for example, a bicyclic heteroaryl having at least one ring nitrogen atom. In some embodiments, W _d is a bicyclic heteroaryl having at least two heteroatoms, for example, a bicyclic heteroaryl having at least two ring nitrogen atoms. In some embodiments, W _d is a bicyclic heteroaryl having two heteroatoms in the ring that is attached to XY. In some embodiments, W _d is a bicyclic heteroaryl having two ring nitrogen atoms in the ring to which XY is attached. In some embodiments, W _d is a bicyclic heteroaryl having four heteroatoms, for example, a bicyclic heteroaryl having four ring nitrogen atoms. In some embodiments, W _d is unsubstituted or substituted 4-amino-1H-pyrazolo [3,4-d] pyrimidin-1-yl, unsubstituted or substituted 7-amino-2-methyl-2H-pyrazolo [4,3-d ] pyrimidin-3-yl, unsubstituted or substituted 6-methylene-9H-purin-6-yl or unsubstituted or substituted 6-amino-9H-purin-9-yl.

In some embodiments, W _d is one of the following:

,

,

where R ^a represents hydrogen, halogen, phosphate, urea, carbonate, alkyl, alkenyl, alkynyl, cycloalkyl, heteroalkyl or heterocycloalkyl;

R ¹¹ represents H, alkyl, halogen, amino, amido, hydroxy or alkoxy, and

R ¹² represents H, alkyl, cyano, alkynyl, alkenyl, halo, aryl, heteroaryl, heterocycloalkyl, cycloalkyl, amino, carboxylic acid, alkoxycarbonyl or amido.

In some embodiments, W _d represents:

.

.

In some embodiments, W _d represents:

.

.

In some embodiments, W _d represents:

.

.

In some embodiments, W _d represents:

.

.

In some embodiments, W _d R ^a is a member selected from the group consisting of hydrogen, halogen, phosphate, urea, carbonate, unsubstituted or substituted alkyl, unsubstituted or substituted alkenyl, unsubstituted or substituted alkynyl, unsubstituted or substituted cycloalkyl, unsubstituted or substituted heteroalkyl; and unsubstituted or substituted heterocycloalkyl.

In some embodiments of W _d , when R ^a is alkyl, alkynyl, cycloalkyl, heteroalkyl or heterocycloalkyl, it is substituted with phosphate, urea or carbonate.

In some embodiments of a compound of formula I, IR ¹¹ is a member of the group consisting of hydrogen, unsubstituted or substituted alkyl, and halo, which includes —I, —F, —Cl, or —Br. In some embodiments, R ¹¹ is unsubstituted or substituted amino, unsubstituted or substituted amido, hydroxy, or unsubstituted or substituted alkoxy. In some embodiments, R ¹¹ is phosphate, unsubstituted or substituted urea or carbonate.

In some embodiments, when R ¹¹ is alkyl, amino, amido, hydroxy, or alkoxy, it is substituted with phosphate, urea, or carbonate.

In some embodiments of the compounds of formula I, -XYW _d represents one of the following groups:

In some embodiments of a compound of formula I, IR ¹² is a member of the group consisting of hydrogen, cyano, halo, unsubstituted or substituted alkyl, and unsubstituted or substituted alkynyl, unsubstituted or substituted alkenyl. In some embodiments, R ¹² is unsubstituted or substituted aryl. In some embodiments, R ¹² is an unsubstituted or substituted heteroaryl, which includes, but is not limited to, heteroaryl having a 5 membered ring, heteroaryl having a six membered ring, heteroaryl with at least one ring nitrogen atom, heteroaryl with two ring nitrogen atoms monocyclic heteroaryl and bicyclic heteroaryl. In some embodiments, R ¹² is an unsubstituted or substituted heterocycloalkyl that includes, but is not limited to, a single ring nitrogen atom, a heterocycloalkyl with one ring oxygen atom, R ¹² represents a single ring heterocycloalkyl group, 5 membered heterocycloalkyl, 6 membered heterocycloalkyl, saturated heterocycloalkyl, unsaturated heterocycloalkyl, heterocycloalkyl having an unsaturated moiety attached to a heterocycloalkyl ring, heterocyclo alkyl substituted with oxo; and heterocycloalkyl substituted with two oxo. In some embodiments, R ¹² is unsubstituted or substituted cycloalkyl, including, but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloalkyl, substituted with one oxo, cycloalkyl having an unsaturated moiety attached to a cycloalkyl ring. In some embodiments, R ¹² is unsubstituted or substituted amido, carboxylic acid, unsubstituted or substituted acyloxy, or unsubstituted or substituted alkoxycarbonyl.

In some embodiments, when R ¹² is alkyl, alkynyl, alkenyl, aryl, heteroaryl, heterocycloalkyl or cycloalkyl, it is substituted with phosphate. In some embodiments, when R ¹² is alkyl, alkynyl, alkenyl, aryl, heteroaryl, heterocycloalkyl or cycloalkyl, it is substituted with urea. In some embodiments, when R ¹² is alkyl, alkynyl, alkenyl, aryl, heteroaryl, heterocycloalkyl or cycloalkyl, it is substituted with carbonate.

In some embodiments, when R ¹² is alkyl, alkynyl, alkenyl, aryl, heteroaryl, heterocycloalkyl, cycloalkyl, alkoxycarbonyl, amido or acyloxy, it is substituted with one or more of alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl , heteroaryl, alkoxy, amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy or nitro, each alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amido, , acyloxy, alkox carbonyl or sulfonamido itself may be substituted.

In some embodiments, R ¹² in W _d represents one of the following groups:

-CN, -Br, -Cl, -I, -H, -Me, -Et, -iso-Pr,

In some embodiments, W _d is pyrazolopyrimidine of formula III:

where R ¹¹ represents H, alkyl, halogen, amino, amido, hydroxy or alkoxy; and R ¹² represents H, alkyl, alkynyl, alkenyl, halo, aryl, heteroaryl, heterocycloalkyl or cycloalkyl. In some embodiments, R ¹¹ is amino; and R ¹² represents H, alkyl, alkynyl, alkenyl, halo, aryl, heteroaryl, heterocycloalkyl or cycloalkyl. In some embodiments, R ¹¹ is amino; and R ¹² represents alkyl, halo, aryl, heteroaryl, heterocycloalkyl or cycloalkyl. In some embodiments, R ¹¹ is amino; and R ¹² is monocyclic heteroaryl. In some embodiments, R ¹¹ is amino; and R ¹² is a bicyclic heteroaryl. In some embodiments, R ¹¹ is amino; and R ¹² is cyano, amino, carboxylic acid, acyloxy, alkoxycarbonyl or amido.

In some embodiments of the invention, the compound of formula I is a compound having the structure of formula IV:

.

.

In some embodiments of the compounds of formula IV, R ¹¹ is H, alkyl, halo, amino, amido, hydroxy or alkoxy; and R ¹² represents H, alkyl, alkynyl, alkenyl, halo, aryl, heteroaryl, heterocycloalkyl or cycloalkyl. In another embodiment, R ¹¹ is amino; and R ¹² represents alkyl, alkenyl, heteroaryl, aryl or heterocycloalkyl. In some embodiments, R ¹¹ is amino; and R ¹² represents cyano, amino, carboxylic acid, alkoxycarbonyl or amido.

In some embodiments, the compound of formula IV is a compound of formula IV-A:

.

.

The invention also provides compounds of formula I having the structure of any of formulas V, V-A, V-B, VI or VI-A:

Any of the described elements and their substituents for compounds of formula I can be used in any combination.

In one aspect, R ³ is H, CH ₃ , CF ₃ , Cl, or F; B is a grouping of formula II:

,

,

where W _c represents aryl, heteroaryl, heterocycloalkyl or cycloalkyl; R ¹ represents H, —F, —Cl, —CN, —CH ₃ , isopropyl, —CF ₃ , —OCH ₃ , nitro or phosphate; R ² represents halogen, hydroxy, cyano or nitro; q is an integer of 0, 1, 2, 3 or 4; R ⁵ , R ⁶ , R ⁷ and R ⁸ are H; X is absent or represents (CH ₂ ) _z ; z is 1; Y is absent or is —N (R ⁹ ) -; R ⁹ represents hydrogen, C ₁ -C ₁₀ alkyl, C ₃ -C ₇ cycloalkyl or C ₂ -C ₁₀ heteroalkyl; and W _d is pyrazolopyrimidine or purine.

или

R¹¹ представляет собой амино; и R¹² представляет собой Н, алкил, алкинил, алкенил, галогено, арил, гетероарил, гетероциклоалкил или циклоалкил.In another aspect, R ³ is H, CH ₃ , CF ₃ , Cl, or F; B is a moiety of formula II, which is aryl, heteroaryl, heterocycloalkyl or cycloalkyl, R ¹ is H, —F, —Cl, —CN, —CH ₃ , isopropyl, —CF ₃ , —OCH ₃ , nitro or phosphate ; R ² represents halogen, hydroxy, cyano or nitro; q is 0, 1 or 2; R ⁵ , R ⁶ , R ⁷ and R ⁸ are H; X is absent or represents (CH ₂ ) _z ; z is 1; Y is absent or is —N (R ⁹ ) -; R ⁹ represents hydrogen, methyl or ethyl; W _d represents:

or

R ¹¹ represents amino; and R ¹² represents H, alkyl, alkynyl, alkenyl, halo, aryl, heteroaryl, heterocycloalkyl or cycloalkyl.

; R¹¹ представляет собой амино; и R¹² представляет собой Н, алкил, алкинил, алкенил, галогено, арил, гетероарил, гетероциклоалкил или циклоалкил.In another aspect, R ³ is H, CH ₃ , CF ₃ , Cl, or F; B is a moiety of formula II, which is aryl, heteroaryl, heterocycloalkyl or cycloalkyl, R ¹ is H, —F, —Cl, —CN, —CH ₃ , isopropyl, —CF ₃ , —OCH ₃ , nitro or phosphate ; R ² represents halogen, hydroxy, cyano or nitro; q is 0, 1 or 2; X represents (CH ₂ ) _z ; z is 1; R ⁵ , R ⁶ , R ⁷ and R ⁸ are H; Y is absent; and W _d represents:

; R ¹¹ represents amino; and R ¹² represents H, alkyl, alkynyl, alkenyl, halo, aryl, heteroaryl, heterocycloalkyl or cycloalkyl.

;In another aspect, R ³ is H, CH ₃ , CF ₃ , Cl, or F; B represents aryl, heteroaryl, heterocycloalkyl or cycloalkyl, R ¹ represents H, —F, —Cl, —CN, —CH ₃ , isopropyl, —CF ₃ , —OCH ₃ , nitro or phosphate; R ² represents halogen, hydroxy, cyano or nitro; q is O, 1 or 2; R ⁵ , R ⁶ , R ⁷ and R ⁸ are H; X represents (CH ₂ ) _z ; z is 1; X represents (CH ₂ ) _z ; z is 1; Y represents —N (R ⁹ ) -; R ⁹ represents hydrogen, methyl or ethyl; and W _d represents

;

R ¹¹ represents amino; and R ¹² represents H, alkyl, alkynyl, alkenyl, halo, aryl, heteroaryl, heterocycloalkyl or cycloalkyl.

In another aspect, R ³ is aryl, heteroaryl, H, CH ₃ , CF ₃ , Cl or F; B is an alkyl or group of formula II:

,

,

where W _c represents aryl, heteroaryl, heterocycloalkyl or cycloalkyl; and q is an integer of 0, 1, 2, 3, or 4; R ¹ represents H, —F, —Cl, —CN, —CH ₃ , isopropyl, —CF ₃ , —OCH ₃ , nitro or phosphate; R ² represents halogen, hydroxy, cyano, nitro or phosphate; q is 0, 1 or 2; R ⁵ , R ⁶ , R ⁷ and R ⁸ are H; X is absent or represents (CH (R ⁹ )) _z ; z is an integer of 1, 2, 3 or 4; Y is absent, represents —N (R ⁹ ) - or —N (R ⁹ ) CH (R ⁹ ) -; R ⁹ represents hydrogen, alkyl, cycloalkyl or heteroalkyl; and W _d is pyrazolopyrimidine or purine.

или

; R¹¹ представляет собой амино; и R¹² представляет собой Н, алкил, алкинил, алкенил, галогено, арил, гетероарил, гетероциклоалкил, циклоалкил, циано, амино, карбоновую кислоту, алкоксикарбонил или амидо.In another aspect, R ³ is aryl, heteroaryl, H, CH ₃ , CF ₃ , Cl or F; B is an alkyl group or a group of formula II, which is aryl, heteroaryl, heterocycloalkyl or cycloalkyl; R ¹ represents H, —F, —Cl, —CN, —CH ₃ , isopropyl, —CF ₃ , —OCH ₃ , nitro or phosphate; R ² represents halogen, hydroxy, cyano, nitro or phosphate; q is 0, 1 or 2; R ⁵ , R ⁶ , R ⁷ and R ⁸ are H; X is absent or represents (CH (R ⁹ )) _z ; z is an integer of 1, 2, 3 or 4; Y is absent, represents —N (R ⁹ ) - or —N (R ⁹ ) CH (R ⁹ ) -; R ⁹ represents hydrogen, methyl or ethyl; W _d represents:

or

; R ¹¹ represents amino; and R ¹² represents H, alkyl, alkynyl, alkenyl, halo, aryl, heteroaryl, heterocycloalkyl, cycloalkyl, cyano, amino, carboxylic acid, alkoxycarbonyl or amido.

; R¹¹ представляет собой амино; и R¹² представляет собой Н, алкил, алкинил, алкенил, галогено, арил, гетероарил, гетероциклоалкил, циклоалкил, циано, амино, карбоновую кислоту, алкоксикарбонил или амидо.In another aspect, R ³ is H, CH ₃ , CF ₃ , Cl, or F; B is an alkyl group or a group of formula II, which is aryl, heteroaryl, heterocycloalkyl or cycloalkyl; R ¹ represents H, —F, —Cl, —CN, —CH ₃ , isopropyl, —CF ₃ , —OCH ₃ , nitro or phosphate; R ² represents halogen, hydroxy, cyano, nitro or phosphate; q is 0, 1 or 2; R ⁵ , R ⁶ , R ⁷ and R ⁸ are H; X represents (CH (R ⁹ )) _z ; z is an integer of 1; Y is absent or represents —N (R ⁹ ) CH (R ⁹ ) -; R ⁹ represents hydrogen, methyl or ethyl; W _d represents:

; R ¹¹ represents amino; and R ¹² represents H, alkyl, alkynyl, alkenyl, halo, aryl, heteroaryl, heterocycloalkyl, cycloalkyl, cyano, amino, carboxylic acid, alkoxycarbonyl or amido.

.In another aspect, R ³ is aryl, heteroaryl, H, CH ₃ , CF ₃ , Cl or F; B is a moiety of formula II, which is aryl, heteroaryl, heterocycloalkyl or cycloalkyl; R ¹ represents H, —F, —Cl, —CN, —CH ₃ , isopropyl, —CF ₃ , —OCH ₃ , nitro or phosphate; R ² represents halogen, hydroxy, cyano, nitro or phosphate; q is 0, 1 or 2; R ⁵ , R ⁶ , R ⁷ and R ⁸ are H; X is absent or represents (CH (R ⁹ )) _z ; z is an integer of 1, 2, 3 or 4; Y is absent, represents —N (R ⁹ ) - or —N (R ⁹ ) CH (R ⁹ ) -; R ⁹ represents hydrogen, methyl or ethyl; and W _d represents:

.

.In another aspect, R ³ is aryl, heteroaryl, H, CH ₃ , CF ₃ , Cl or F; B is a moiety of formula II, which is aryl, heteroaryl, heterocycloalkyl or cycloalkyl; R ¹ represents H, —F, —Cl, —CN, —CH ₃ , isopropyl, —CF ₃ , —OCH ₃ , nitro or phosphate; R ² represents halogen, hydroxy, cyano, nitro or phosphate; q is 0, 1 or 2; R ⁵ , R ⁶ , R ⁷ and R ⁸ are H; X is absent; Y represents —N (R ⁹ ) CH (R ⁹ ) -; R ⁹ represents hydrogen, methyl or ethyl; and W _d represents:

.

Additional representative compounds of the present invention having a substructure of formula IV-A are described.

.

.

Some illustrative compounds of the present invention having the structure of formula IV-A include compounds in which R ³ is —H, —Cl, —F, or —CH ₃ , in combination with any moiety B described in Table 1, and any R ¹² , as described in Table 2. The compound of formula IV-A includes any combination of R ³ , B and R ¹² . Additional representative compounds of formula IV-A are illustrated in Table 4.

Table 1 Illustrative Grouping B in the Compounds of Formula I No. under
class AT No. under
class AT No. under
class AT IN 1

IN 2

IN 3 -CH (CH ₃ ) ₂ AT 4

AT 5

AT 6

AT 7

AT 8

AT 9

AT 10

AT 11

AT 12

B-13

B-14

B-15

B-16

B-17

B-18

B-19

IN 20

AT 21

B-22

B-23

B-24

B-25

B-26

B-27

B-28

B-29

B-30

B-31

B-32

B-33

B-34

B-35

B-36

B-37

B-38

B-39

B-40

B-41

B-42

B-43

B-44

B-45

B-46

B-47

B-48

B-49

B-50

B-51

B-52

B-53

B-54

B-55

B-56

B-57

B-58

B-59

B-60

B-61

B-62

B-63

B-64

B-65

B-66

B-67

B-68

B-69

B-70

B-71

B-72

B-73

B-74

B-75

B-76

B-77

B-78

B-79

B-80

B-81

B-82

B-83

B-84

B-85

B-86

B-87 -CH ₃ B-88 -CH ₂ CH ₃ B-89

B-90

B-91

B-92

B-93

B-94

B-95

B-96

B-97

B-98

B-99

B-100

B-101

B-102

table 2 Illustrative Examples of R ¹² in Compounds of Formula I No.
class R ¹² No.
class R ¹² No.
class R ¹² 12-1 -CN 12-2 -Br 12-3 -Cl 12-4 -CH ₂ CH ₃ 12-5 -CH ₃ 12-6 -CH (CH ₃ ) ₂ 12-7

12-8

12-9

12-10

12-11

12-12

12-13

12-14

12-15

12-16

12-17

12-18

12-19

12-20

12-21

12-22

12-23

12-24

12-25

12-26

12-27

12-28

12-29

12-30

12-31

12-32

12-33

12-34

12-35 -H 12-36

12-37

12-38

12-39

12-40

12-41

12-42

12-43

12-44

12-45

12-46

2-47

2-48

12-49

12-50

12-51

12-52

12-53

12-54

12-55

12-56

12-57

12-58

12-59

12-60

12-61 -I 12-62

12-63

12-64

12-65

12-66

12-67

12-68

12-69

12-70

12-71

12-72

12-73

12-74

12-75

12-76

12-77

12-78

12-79

12-80

12-81

12-82

12-83

12-84

12-85

12-86

12-87

12-88

12-89

12-90

12-91

12-92

12-93

12-94

12-95

12-96

Other exemplary compounds of the present invention have the structure of formula VA wherein B is a moiety described in Table 2 in combination with R ³ which is —H, —Cl, —F, or —CH ₃ ; and R ⁹ , which is —H, —CH ₃ or —CH ₂ CH ₃ .

.

.

Other illustrative compounds of the present invention have the structure of formula VB in which B is a moiety described in Table 2 in combination with R ³ which is —H, —Cl, —F, or —CH ₃ ; and R ⁹ , which is —H, —CH ₃ or —CH ₂ CH ₃ .

Other illustrative compounds of the present invention have the structure of formula VI-A, in which B is a moiety described in Table 2, in combination with R ³ , which is —H, —Cl, —F, or —CH ₃ ; and R ⁹ , which is —H, —CH ₃ or —CH ₂ CH ₃ .

Additional typical compounds of the present invention include, but are not limited to, the following:

.

.

The chemical structures set forth in this application can be synthesized in accordance with one or more illustrative schemes in this application and / or methods well known in the art.

Unless otherwise indicated, the reactions described in this application proceed at atmospheric pressure, in the General case in the temperature range from -10 ° C to 200 ° C. Moreover, unless otherwise indicated, it is understood that the duration of the reactions and conditions are indicated approximately, for example, they occur at about atmospheric pressure in the temperature range from about -10 ° C to about 110 ° C for a period of time from about 1 up to about 24 hours; the duration of the reactions left overnight is on average about 16 hours.

The terms “solvent”, “organic solvent” or “inert solvent” each mean a solvent inert under the conditions of the reaction described herein, including, for example, benzene, toluene, acetonitrile, tetrahydrofuran (“THF”), dimethylformamide (“DMF”), chloroform, methylene chloride (or dichloromethane (DCM)), diethyl ether, methanol, N-methylpyrrolidone ("NMP"), pyridine and the like. Unless otherwise indicated, the solvents used in the reactions set forth in this application are inert organic solvents. Unless otherwise indicated, for each gram of limiting reagent, the equivalent volume is one cm ³ (ss) (or ml) of solvent.

If desired, the isolation and purification of the chemical structures and intermediates set forth in this application can be carried out using any suitable separation or purification technique, such as, for example, filtration, extraction, crystallization, column chromatography, thin layer chromatography or thick layer chromatography or a combination these techniques. Specific illustrations of suitable separation and isolation techniques can be obtained by reference to the examples below. However, other equivalent separation or isolation techniques may also be used.

If desired, the (R) and (S) -isomers of the compounds of the present invention, if present, can be separated by methods known to those skilled in the art, for example, by forming diastereoisomeric salts or complexes that can be separated, for example crystallization; through the formation of diastereoisomeric derivatives, which can be separated, for example, by crystallization, gas-liquid or liquid chromatography; by selectively reacting one enantiomer with an enantiomer-specific reagent, for example, enzymatic oxidation or reduction, followed by separation of the modified and unmodified enantiomers; or by gas-liquid or liquid chromatography in a chiral environment, for example, on a chiral support, such as silica with an attached chiral ligand, or in the presence of a chiral solvent. Alternatively, a particular enantiomer can be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer to another by asymmetric transformation.

The compounds described herein may possibly be contacted with a pharmaceutically acceptable acid to form the corresponding acid addition salts.

Most of the possibly substituted starting compounds and other reagents are commercially available, for example from Aldrich Chemical Company (Milwaukee, WI), or can be readily prepared by those skilled in the art using a commonly used synthesis methodology.

In general, the compounds of the invention can be synthesized using an appropriate combination of common well-known synthetic methods. Techniques useful in the synthesis of these chemical structures are both readily understood and available to those skilled in the relevant field of technology in view of the present description.

The compounds of the invention can be synthesized using an appropriate combination of synthetic methods known in the art. The following discussion is provided to illustrate some of the various methods available for use in the preparation of compounds of the invention, and is not intended to limit the scope of reactions or reaction sequences that can be used in the preparation of compounds of the present invention.

Reaction scheme 1:

According to Scheme 1, step 1, a compound of formula 101, where X is N or CR ⁷ , is converted to a compound of formula 103, for example, through a two-step process for combining Heck with a compound of formula 102 followed by acid catalyzed cyclization in methanol. The product, a compound of formula 103, is isolated. According to Scheme 1, step 2, a compound of formula 103 is converted to a compound of formula 104, for example, by reaction with a suitably substituted aniline. The product, a compound of formula 104, is isolated. According to Scheme 1, step 3, a compound of formula 104 is converted to a compound of formula 105, for example, by reduction using lithium aluminum hydride. The product, a compound of formula 105, is isolated. According to Scheme 1, step 4, a compound of formula 105 is converted to a compound of formula 106, for example, by reaction with thionyl chloride. The product, a compound of formula 106, is isolated. According to Scheme 1, step 5, a compound of formula 106 is converted to a compound of formula 107, for example, by alkylation with pyrazolopyrimidine using a base such as potassium carbonate. The product, a compound of formula 107, is isolated. According to Scheme 1, step 6, a compound of formula 107 is converted to a compound of formula 108, for example, by means of a Suzuki reaction. The product, a compound of formula 108, is isolated and further purified.

Reaction Scheme 2:

According to Scheme 2, step 1, a compound of formula 201, where X is N or CR ⁷ , is converted to a compound of formula 202, for example, using a reagent suitable for the administration of an acid chloride, for example, oxalyl chloride. The product, a compound of formula 202, is possibly isolated. According to Scheme 2, stage 2, the compound of formula 202 is converted into a compound of formula 203, for example, by reaction, for example, with arylamine. The product, a compound of formula 203, is isolated. According to Scheme 2, step 3, a compound of formula 203 is converted into a compound of formula 204, for example, by means of a Stille combination using the appropriate vinyl stannane. The product, a compound of formula 204, is isolated. According to Scheme 2, step 4, a compound of formula 204 is converted to a tertiary amide, a compound of formula 205, by reaction with chloroethyl acetate and base with sodium hydride. The compound of formula 205 is isolated. According to Scheme 2, step 5, the compound of formula 205 is oxidized to an aldehyde using, for example, osmium tetroxide and sodium periodinate. The product, a compound of formula 206, is isolated. According to Scheme 2, step 6, a compound of formula 206 is converted to a compound of formula 104, for example, by aldol reaction in ethanol in the presence of a base such as cesium carbonate. The product, a compound of formula 104, is isolated. According to Scheme 2, step 7, a compound of formula 104 is reduced to a primary alcohol by reduction using, for example, lithium aluminum hydride, to give a compound of formula 105 that is isolated. According to Scheme 2, step 8, a compound of formula 105 is converted to a compound of formula 207 by reaction with carbon tetrabromide and triphenylphosphine. The compound of formula 207 is isolated. This compound may be a central intermediate in the synthesis of compounds of the invention.

Reaction Scheme 3:

According to Scheme 3, stage 9, a compound of formula 207, where X is N or CR ⁷ , synthesized as described in Reaction scheme 2, and converted into a compound of formula 107 by combination with a compound of formula 208 in the presence of a base, for example, tert-butylate potassium. The compound of formula 107 is isolated. According to Scheme 3, step 10, a compound of formula 107 is converted to a compound of formula 108 by coupling, for example, with arylboronic acid in the presence of coupling and base reaction catalysts, for example palladium acetate, triphenylphosphine and sodium carbonate, for example. The compound of formula 108 is isolated.

Reaction scheme 4:

According to Reaction Scheme 4, Step 1, an iodine ester 401 is reacted with an alkyn in the presence of a palladium catalyst, copper iodide and triethylamine (TEA) to attach this alkyn to the aryl core of compound 401 to give a compound of formula 402. A compound of formula 402 may be isolated . According to Reaction Scheme 4, Step 2, a compound of formula 402 is treated with a base with potassium hydroxide to give a carboxylic acid, a compound of formula 403 if the reaction product is acidified, or a salt thereof. The compound of formula 403 is possibly isolated. According to Reaction Scheme 4, Step 3, the compound of formula 403 is treated with bis (acetonitrile) dichloropalladium (H) and TEA to effect an intramolecular ring closure to obtain a compound of formula 404. The compound of formula 404 is isolated. According to Reaction Scheme 4, Step 4, a compound of formula 404 is reacted with a primary amine to give a compound of formula 405. A compound of formula 405 is optionally isolated. According to Reaction Scheme 4, Step 5, the compound of formula 405 is treated with hydrochloric acid, removing the protecting group on nitrogen and obtaining the compound of formula 406. The compound of formula 406 is possibly isolated. According to Reaction Scheme 4, Step 6, a compound of formula 406 is reacted with a compound of formula 407 to give a compound of formula 408. The compound of formula 408 is isolated.

Reaction scheme 5:

According to Reaction Scheme 5, Step 1, the iodine ester 401 is reacted with alkyn 501 in the presence of a palladium catalyst of the coupling reaction, copper iodide and TEA to give a compound of formula 502. A compound of formula 502 may be isolated. According to Reaction Scheme 5, Step 2, the compound of Formula 502 is treated with a base with potassium hydroxide to give the carboxylate or free acid of the compound of Formula 503. According to Reaction Scheme 5, Step 3, the compound of Formula 503 is treated with bis (acetonitrile) -dichloropalladium (II) and TEA to effect intramolecular ring closure to obtain a compound of formula 504. A compound of formula 504 may be isolated. According to Reaction Scheme 5, Step 4, a compound of formula 504 is treated with a primary amine to give a compound of formula 505. A compound of formula 505 is isolated.

Reaction scheme 6:

According to Reaction Scheme 6, Step 1, an iodine ester 401 is reacted with Alkine 601 in the presence of a palladium catalyst of the coupling reaction, copper iodide and TEA to give a compound of formula 602. A compound of formula 602 is optionally isolated. According to Reaction Scheme 6, step 2, a compound of formula 602 is treated with a base with potassium hydroxide to give the carboxylate or free acid of a compound of formula 603. According to Reaction Scheme 6, step 3, a compound of formula 603 is treated with bis (acetonitrile) -dichloropalladium (II) and TEA to effect intramolecular ring closure to obtain a compound of formula 604. A compound of formula 604 may be isolated. According to Reaction Scheme 6, Step 4, a compound of formula 604 is treated with a primary amine to give a compound of formula 605. A compound of formula 605 is isolated. According to Reaction Scheme 6, Step 5, the compound of formula 605 is treated with acid to remove the THP protecting group to give the compound of formula 606. The compound of formula 606 is isolated.

Reaction Scheme 7:

According to Reaction Scheme 7, Step 1, a compound of Formula 701 is synthesized by various synthesis methods, including those of Schemes 1 or 2, where, for example, benzylamine is used in the step of converting a compound of Formula 103 to a compound of Formula 104. The benzyl amine protecting group may be removed in accordance with standard chemical deprotection reactions to give compound 701. According to another example of the conversion of a compound of formula 103 to a compound of formula 701, a compound of formula 701 is obtained by treating the compound of formula 103 with ammonia. The compound of formula 701 is converted to the compound of formula 702 by alkylation of amide nitrogen using a series of 2-carbon synthons which can be subjected to deprotection, oxidation and re-protection of the corresponding ketal to the compound of formula 702. According to Reaction Scheme 7, Step 2- 1, a compound of formula 702 is transformed, for example, by reductive amination of an ester moiety to introduce a purinyl group of a compound of formula 703 or, alternatively, alkylated for the purpose of introducing a purinyl group and preparing a compound of formula 703. According to Reaction Scheme 7, Step 3-1, a compound of formula 703 is treated with an acid to remove a ketal protecting group to give a compound of formula 704. A compound of formula 704 is isolated. According to Reaction Scheme 7, Step 4-1, a compound of formula 704 is subjected to reductive amination using an amine to give a compound of formula 705. A compound of formula 705 is isolated. According to Reaction Scheme 7, Steps 2-2, the compound of Formula 702 is converted in accordance with Steps 7 and 8 of Scheme 2 and Step 9 of Scheme 3 to introduce a pyrazolopyrimidine moiety of the compound of Formula 706. The compound of Formula 706 is isolated. According to Reaction Scheme 7, Steps 3-2, a compound of formula 706 is treated with an acid to remove a ketal protecting group to give a compound of formula 707. The compound of formula 707 is isolated. According to Reaction Scheme 7, Steps 4-2, a compound of formula 707 is subjected to reductive amination using an amine to give a compound of formula 708. The compound of formula 708 is isolated.

Reaction Scheme 8:

According to Reaction Scheme 8, step 1, the compound of formula 701 is synthesized as described in Scheme 7, or by any other well-known chemical reactions. The compound of formula 701 is converted by alkylation of amide nitrogen using a series of 2-carbon-containing synthons that can be subjected to deprotection and converted to an alkoxy protected species, as shown for the compound of formula 801, which can be isolated. According to Reaction Scheme 8, Step 2, the compound of Formula 801 is converted using the chemical reactions described in Step 2-1 of Scheme 7 to introduce a purinyl moiety, and such a resulting compound is converted by deprotection, activation and amination using an amine to give a compound of the formula 802, which is allocated.

According to Reaction Scheme 8, Step 3, a compound of Formula 801 is converted using the chemical reactions described in Step 2-2 of Scheme 7 to introduce a pyrazolopyrimidine moiety, and such a resulting compound is converted by deprotection, activation and amination using an amine to give a compound of formula 803, which is isolated.

Reaction scheme 9:

According to Reaction Scheme 9, Step 1, a compound of formula 901 is treated with an amine to give a compound of formula 902. A compound of formula 902 is isolated. According to Reaction Scheme 9, Step 2, a compound of Formula 902 is treated with phosphorus oxychloride to form a compound of Formula 903. A compound of Formula 903 is isolated. According to Reaction Scheme 9, Step 3, a compound of formula 903 is reacted with aminopurine of formula 904 to give a compound of formula 905. A compound of formula 905 is isolated. According to Reaction Scheme 9, step 4, the compound of formula 905 is treated with hydrochloric acid to remove the nitrogen protecting group of the purine moiety to give a compound of formula 906. The compound of formula 906 is isolated.

The reaction scheme 10:

According to Reaction Scheme 10, Step 1, a compound of formula 1001 is treated with vinyl ester 1002 using, for example, a Heck reaction followed by cyclization to give a compound of formula 1003. A compound of formula 1003 is isolated. According to Reaction Scheme 10, Step 2, a compound of formula 1003 is reacted with 4-amino-N-Boc-piperidine to give a compound of formula 1004. A compound of formula 1004 is isolated. The compound of formula 1004 can be used as an intermediate in the synthesis of compounds of the invention.

The reaction scheme 11:

According to Reaction Scheme 11, step 1, the compound of formula 1101 is treated with alkynyl alcohol, for example, of formula 1102, in the presence of copper iodide and a palladium-carbon catalyst to give a compound of formula 1103. The compound of formula 1103 is isolated. According to Reaction Scheme 11, Step 1, a compound of formula 1102 is reacted with 4-amino-N-Boc-piperidine to give a compound of formula 1103. A compound of formula 1103 is isolated. The compound of formula 1103 can be used as an intermediate in the synthesis of compounds of the invention.

The compounds of formula I can be synthesized using the reaction schemes described in this application, or variants of these methods.

These chemical structures can be synthesized using an appropriate combination of common well-known synthetic methods.

In some embodiments, the compounds of the present invention exhibit one or more of the functional characteristics disclosed herein. For example, one or more compounds of the invention specifically bind to PI3 kinase. In some embodiments, the IC50 for the compound of the invention with respect to p110α, p110β, p110γ or p110δ is less than about 1 μM, less than about 100 nM, less than about 50 nM, less than about 10 nM, less than about 1 nM less than about 0.5 nM, less than about 100 pM, or less than about 50 pM.

In some embodiments, one or more of the compounds of the invention can selectively inhibit one or more members of type I or class I phosphatidylinositol-3-kinases (PI3 kinases) with an IC50 value of about 100 nM, 50 nM, 10 nM, 5 nM, 100 pM, 10 pM or 1 pM or less, as measured in an in vitro kinase assay.

In some embodiments, one or more of the compounds of the invention can selectively inhibit one or two kinases from phosphatidylinositol-3-kinases (PI3 kinases) of type I or class I, consisting of PI3 kinase α, PI3 kinase β , PI3 kinases γ and PI3 kinases δ. In some aspects, some of the compounds of the invention selectively inhibit PI3 kinase 5 compared to all other Type I PI3 kinases. In other aspects, some of the compounds of the invention selectively inhibit PI3 kinase δ and PI3 kinase γ compared to other Type I PI3 kinases. In yet other aspects, some of the compounds of the invention selectively inhibit PI3 kinase α and PI3 kinase δ compared to other Type I PI3 kinases. In yet some other other aspects, some of the compounds of the invention selectively inhibit PI3 kinase 6 and PI3 kinase α as compared to other Type I PI3 kinases. In still some other aspects, some of the compounds of the invention selectively inhibit PI3 kinase 6 and PI3 kinase β compared to other Type I kinases, or selectively inhibit PI3 kinase δ and PI3 kinase α compared to the remaining PI3 kinases of type I either selectively inhibit the PI3 kinase α and PI3 kinase γ compared to the remaining PI3 kinases of I or selectively inhibit the PI3 kinase γ and PI3 kinase β compared to other PI3 kinases of the type .

In yet another aspect, an inhibitor that selectively inhibits one or more kinases from Type I PI3 kinases, or an inhibitor that selectively inhibits one or more of Type I PI3 kinases that mediates signal transduction pathways, can alternatively be understood as referring to a compound which exhibits a 50% inhibition concentration (IC50) in relation to a given Type I PI3 kinase that is at least 10 times lower, at least 20 times, at least 50 times, at least 100 times at least 1000 times at least Leray in 10100 times or below the IC50 for the inhibitor for the remaining other PI3 kinase I type.

Pharmaceutical Compositions

The present invention provides pharmaceutical compositions comprising one or more compounds of the present invention.

In some embodiments, the present invention provides pharmaceutical compositions for treating diseases or conditions associated with an undesirable, overactive, harmful or damaging immune response in a mammal. Such an undesirable immune response may be associated with or may cause, for example, asthma, emphysema, bronchitis, psoriasis, allergies, anaphylaxis, autoimmune diseases, rheumatoid arthritis, graft versus host disease and lupus erythematosus. The pharmaceutical compositions of the present invention can be used to treat other respiratory diseases, including, but not limited to, diseases affecting the lobes of the lung, pleural cavity, bronchioles, trachea, upper respiratory tract or nerves and muscles involved in the act of breathing.

In some embodiments, the invention provides pharmaceutical compositions for treating disorders such as hyperproliferative disorder, including, but not limited to, cancer, such as acute myeloid leukemia, cancer of the thymus, brain, lung, squamous cell, skin, eye, retinoblastoma, intraocular melanoma, cancer of the oral cavity and oropharynx, bladder, gastrointestinal tract, stomach, pancreas, bladder, breast, cervix, head, neck, renal sphere, kidney, liver, ovary, prostate, colon, esophagus, testicle, gynecological sphere, thyroid gland, central nervous system (CNS), PNS (peripheral nervous system), AIDS-associated (e.g. Kaposi's lymphoma and Kaposi's sarcoma) or virus-induced cancer. In some embodiments, said pharmaceutical composition is intended for the treatment of benign hyperproliferative disorder, such as benign skin hyperplasia (e.g., psoriasis), repeated stenosis or benign hyperproliferative disorder of the prostate (e.g., benign prostatic hypertrophy (BPH)).

The invention also provides compositions for treating liver diseases (including diabetes), pancreatitis or kidney disease (including proliferative glomerulonephritis and diabetes-induced kidney disease) or pain in a mammal.

Moreover, the invention provides a composition for preventing implantation of blastocytes in a mammal.

The invention also relates to a composition for treating a disease related to vasculogenesis or angiogenesis in a mammal, which can manifest itself as tumor angiogenesis, chronic inflammatory disease such as rheumatoid arthritis, inflammatory bowel disease, atherosclerosis, skin diseases such as psoriasis, eczema and scleroderma diabetes, diabetic retinopathy, premature retinopathy, age-related macular degeneration, hemangioma, glioma, melanoma, Kaposi’s sarcoma and ovarian cancer , breast, lungs, pancreas, prostate, colon and epidermoid cancer.

The pharmaceutical compositions of the invention are typically formulated containing a therapeutically effective amount of a compound of the present invention as an active ingredient or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof. Where desired, the pharmaceutical compositions contain a pharmaceutically acceptable salt and / or its coordination complex and one or more pharmaceutically acceptable excipients, carriers, including inert solid diluents and excipients, diluents, including sterile aqueous solution and various organic solvents, substances that increase permeability, solubilizers and adjuvants.

The pharmaceutical compositions of the invention may be administered alone or in combination with one or more other agents, which are usually administered in the form of pharmaceutical compositions. Where desired, the compounds of the invention and the other agent (s) can be mixed in one preparation or both components can be prepared as separate preparations for use in combination individually or at the same time.

In some embodiments, the concentration of one or more compounds proposed in the pharmaceutical compositions of the present invention is less than 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18 %, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0, 05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002% or 0.0001% mass / mass, mass / about. or vol./about.

In some embodiments, the concentration of one or more compounds of the present invention is more than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19.75%, 19.50%, 19.25 % 19%, 18.75%, 18.50%, 18.25% 18%, 17.75%, 17.50%, 17.25% 17%, 16.75%, 16.50%, 16, 25% 16%, 15.75%, 15.50%, 15.25% 15%, 14.75%, 14.50%, 14.25% 14%, 13.75%, 13.50%, 13 , 25% 13%, 12.75%, 12.50%, 12.25% 12%, 11.75%, 11.50%, 11.25% 11%, 10.75%, 10.50%, 10.25% 10%, 9.75%, 9.50%, 9.25% 9%, 8.75%, 8.50%, 8.25% 8%, 7.75%, 7.50% , 7.25% 7%, 6.75%, 6.50%, 6.25% 6%, 5.75%, 5.50%, 5.25% 5%, 4.75%, 4.50 %, 4.25%, 4%, 3.75%, 3.50%, 3.25%, 3%, 2.75%, 2.50%, 2.25%, 2%, 1.75% , 1.50%, 125%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07 %, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002% or 0 , 0001% wt. / W, w / v. or vol./about.

In some embodiments, the concentration of one or more compounds of the present invention is in the range from about 0.0001% to about 50%, from about 0.001% to about 40%, from about 0.01% to about 30%, from about 0.02 % to about 29%, from about 0.03% to about 28%, from about 0.04% to about 27%, from about 0.05% to about 26%, from about 0.06% to about 25%, from about 0.07% to about 24%, from about 0.08% to about 23%, from about 0.09% to about 22%, from about 0.1% to about 21%, from about 0.2% to about 20%, from about 0.3% to about 19%, from about 0 , 4% to about 18%, from about 0.5% to about 17%, from about 0.6% to about 16%, from about 0.7% to about 15%, from about 0.8% to about 14 %, from about 0.9% to about 12%, from about 1% to about 10% w / w, w / v. or vol./about. about / about

In some embodiments, the concentration of one or more compounds of the present invention is in the range from about 0.001% to about 10%, from about 0.01% to about 5%, from about 0.02% to about 4.5%, from about 0 , 03% to about 4%, from about 0.04% to about 3.5%, from about 0.05% to about 3%, from about 0.06% to about 2.5%, from about 0.07 % to about 2%, from about 0.08% to about 1.5%, from about 0.09% to about 1% , from about 0.1% to about 0.9% mass / mass, mass / vol. or vol./about.

In some embodiments, the amount of one or more compounds of the present invention is equal to or less than 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0.3 g, 0.25 g, 0.2 g, 0.15 g, 0.1 g, 0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g, 0.03 g, 0.02 g, 0.01 g, 0.009 g, 0.008 g, 0.007 g , 0.006 g, 0.005 g, 0.004 g, 0.003 g, 0.002 g, 0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006 g, 0.0005 g, 0.0004 g, 0 , 0003 g, 0.0002 g or 0.0001 g.

In some embodiments, the amount of one or more compounds of the present invention is more than 0.0001 g, 0.0002 g, 0.0003 g, .0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g, 0 , 0008 g, 0.0009 g, 0.001 g, 0.0015 g, 0.002 g, 0.0025 g, 0.003 g, 0.0035 g, 0.004 g, 0.0045 g, 0.005 g, 0.0055 g, 0.006 g, 0.0065 g, 0.007 g, 0.0075 g, 0.008 g, 0.0085 g, 0.009 g, 0.0095 g, 0.01 g, 0.015 g, 0.02 g, 0.025 g, 0.03 g, 0.035 g, 0.04 g, 0.045 g, 0.05 g, 0.055 g, 0.06 g, 0.065 g, 0.07 g, 0.075 g, 0.08 g, 0.085 g, 0.09 g, 0.095 g, 0.1 g, 0.15 g, 0.2 g, 0.25 g, 0.3 g, 0.35 g, 0.4 g, 0.45 g, 0.5 g, 0, 55 g, 0.6 g, 0.65 g, 0.7 g, 0.75 g, 0.8 g, 0.85 g, 0.9 g, 0.95 g, 1 g, 1.5 g 2 g, 2.5, 3 g, 3.5, 4 g, 4.5 g, 5 g, 5.5 g, 6 g, 6.5 g, 7 g, 7.5 g, 8 g, 8.5 g, 9 g, 9.5 g or 10 g.

In some embodiments, the amount of one or more compounds of the present invention is in the range of 0.0001-10 g, 0.0005-9 g, 0.001-8 g, 0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g, 0.5-4 g or 1-3 g.

The compounds of the invention are effective over a wide range of doses. For example, in the treatment of an adult, examples of dosages that can be used are dosages of 0.01-1000 mg; 0.5-100 mg; 1-50 mg per day and 5-40 mg per day. A typical dosage is 10-30 mg per day. The exact dosage will depend on the route of administration, the form in which the compound is administered, the subject being treated, the body weight of the subject being treated, and the preference and experience of the attending physician.

Non-limiting typical pharmaceutical compositions and methods for their manufacture are described below.

Pharmaceutical compositions for oral administration. In some embodiments of the present invention, there is provided a pharmaceutical composition for oral administration comprising a compound of the present invention and a pharmaceutical excipient suitable for oral administration.

In certain embodiments of the invention, there is provided a solid pharmaceutical composition for oral administration comprising: (1) an effective amount of a compound of the present invention; possibly (2) an effective amount of a second agent; and (3) a pharmaceutical excipient suitable for oral administration. In some embodiments, the composition further comprises: (4) an effective amount of a third agent.

In some embodiments, the pharmaceutical composition may be a liquid pharmaceutical composition suitable for oral administration. The pharmaceutical compositions of the invention suitable for oral administration can be presented in the form of discrete dosage forms, such as capsules, sachets or tablets, or liquids or aerosol sprays, each of which contains a predetermined amount of the active ingredient in powder form or in granules, solution or suspensions in an aqueous or anhydrous liquid, an oil-in-water emulsion or a water-in-oil liquid emulsion. Such dosage forms can be manufactured by any of the methods known in the pharmaceutical art, however, all methods include the step of bringing the active ingredient into association with a carrier, which is one or more than one necessary ingredient. In general, compositions are made by continuously and uniformly mixing the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product to the desired appearance. For example, a tablet may be made by compression or molding, possibly with one or more than one auxiliary ingredient. Compressed tablets can be made by compressing in a suitable machine the active ingredient in a free-flowing form, such as a powder or granules, possibly mixed with an excipient such as a binder, lubricant, inert diluent and / or surfactant or dispersant, but not limited to this. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

Moreover, the present invention encompasses anhydrous pharmaceutical compositions and dosage forms containing the active ingredient, since water may contribute to the destruction of certain compounds. For example, in the pharmaceutical field, water can be added (for example, 5%) as a tool for modeling long-term storage in order to determine characteristics such as shelf life or stability of compositions over time. Anhydrous pharmaceutical compositions and dosage forms of the invention can be formulated using anhydrous or low moisture ingredients and low moisture or low humidity conditions. The pharmaceutical compositions and dosage forms of the invention that contain lactose can be made anhydrous if substantial contact with moisture and / or moisture is expected during manufacturing, packaging and / or storage. An anhydrous pharmaceutical composition can be prepared and stored in such a way as to maintain its anhydrous state. Accordingly, anhydrous compositions can be packaged using materials known for protecting against ingress of water, such as may be included in suitable kits defined in the pharmacological guide. Examples of suitable packaging include, but are not limited to, foil, plastic, or the like for hermetic closure, disposable containers, blister packs, and tape-type packagings.

The active ingredient can be combined in a homogeneous mixture with a pharmaceutical carrier in accordance with traditional methods of preparing pharmaceutical mixtures. The carrier may take a wide variety of forms, depending on the form of the preparation desired for administration. In the manufacture of oral dosage unit compositions, any conventional pharmaceutical media may be used as carriers, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like in the case of oral liquid preparations (such as suspensions , solutions and elixirs) or aerosols; or carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders and disintegrants, can be used in the case of oral solid preparations, in some embodiments without the use of lactose. For example, suitable carriers include powders, capsules, and tablets containing solid oral preparations. If necessary, tablets may be coated using standard aqueous or anhydrous techniques.

Binders suitable for use in pharmaceutical compositions and dosage forms include, but are not limited to, corn starch, potato starch or other starches, gelatin, natural and synthetic resins such as gum arabic, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g. ethyl cellulose, cellulose acetate, carboxymethyl cellulose calcium salt, carboxymethyl cellulose sodium salt), polyvinylpyrro lidone (PVP), methyl cellulose, pregelatinized starch, hydroxypropyl methyl cellulose, microcrystalline cellulose, and mixtures thereof.

Examples of suitable excipients for use in the pharmaceutical compositions and dosage forms set forth herein include, but are not limited to, talc, calcium carbonate (e.g., in granules or powder), microcrystalline cellulose, powdered cellulose, dextrans, kaolin, mannitol, silicon acid, sorbitol, starch, pregelatinized starch and mixtures thereof.

In the compositions of the invention, disintegrants may be used to prepare tablets which disintegrate when placed in an aqueous environment. If there is too much baking powder, tablets that can disintegrate in a bottle can be made during manufacture. Too small an amount may not be sufficient for degradation to occur, and thus may alter the rate and extent of release of the active ingredient (s) from the dosage form. Thus, for the formation of dosage forms of the compounds described in this application, a sufficient amount of disintegrant can be used, that is, not too small and not too large so as not to adversely affect the release of the active ingredient (s). The amount of disintegrant used may vary depending on the type of composition and route of administration, and can be readily estimated by one of ordinary skill in the art. From the pharmaceutical composition may be used from about 0.5 to about 15 weight percent of baking powder or from about 1 to about 5 weight percent of baking powder. Baking powder that can be used to form the pharmaceutical compositions and dosage forms of the invention include, but are not limited to, agar agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, potassium polacryline, sodium starch glycolate, starch from potatoes or tapioca, other starches, pregelatinized starch, other starches, clays, other algins, other celluloses, resins, or mixtures thereof.

Lubricants that can be used to form pharmaceutical compositions and dosage forms of the invention include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol (PEG), other glycols , stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g. peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil and soybean oil), ste rat zinc, ethyl oleate, ethyl laurate, agar, or mixtures thereof. Additional lubricants include, for example, silica gel siloid, coagulated aerosol synthetic silicon dioxide, or mixtures thereof. A lubricant may optionally be added in an amount of less than about 1 weight percent of the pharmaceutical composition.

If aqueous suspensions and / or elixirs for oral administration are desired, then the main active ingredient for them may be combined with various sweeteners or flavoring agents, coloring agent or colorants and, if necessary, emulsifying and / or suspending agents, together with such diluents, like water, ethanol, propylene glycol, glycerin, and various combinations thereof.

The tablets may be uncoated or coated using known methods to slow disintegration and absorption in the gastrointestinal tract and thereby provide continuous action over a longer period of time. For example, you can use such a time-delaying substance as glyceryl monostearate or glyceryl distearate. Compositions for oral administration can also be presented in the form of hard gelatin capsules, where the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or in the form of soft gelatin capsules, where the active ingredient is mixed with an aqueous or oily medium, for example, peanut butter, liquid paraffin, or olive oil.

Surfactants that can be used to form the pharmaceutical compositions and dosage forms of the invention include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof. That is, a mixture of hydrophilic surfactants can be used, a mixture of lipophilic surfactants can be used, or a mixture of at least one hydrophilic surfactant and at least one lipophilic surfactant can be used.

A suitable hydrophilic surfactant can usually have an HLB of at least 10, while suitable lipophilic surfactants can usually have an HLB of equal to or less than about 10. The empirical parameter used to characterize the relative hydrophilicity and hydrophobicity of nonionic amphiphilic compounds is the hydrophilic-lipophilic balance ("HLB" value). Surfactants with lower HLBs are more lipophilic or hydrophobic and have a higher solubility in oils, while surfactants with higher HLBs are more hydrophilic and have a higher solubility in aqueous solutions. In general, it is believed that hydrophilic surfactants are those compounds that have an HLB value greater than about 10, as well as anionic, cationic or zwitterionic compounds for which the HLB scale is not commonly used. Similarly, lipophilic (i.e. hydrophobic) surfactants are compounds having an HLB value equal to or less than about 10. However, the HLB value of a surfactant is only an approximate guideline commonly used in the manufacture of pharmaceutical and cosmetic emulsions in industry.

Hydrophilic surfactants can be either ionic or non-ionic. Suitable ionic surfactants include, but are not limited to, alkylammonium salts; fusidic acid salts; derivatives of fatty acids and amino acids, oligopeptides and polypeptides; derivatives of glycerides and amino acids, oligopeptides and polypeptides; lecithins and hydrogenated lecithins; lysolecithins and hydrogenated lysolecithins; phospholipids and their derivatives; lysophospholipids and their derivatives; salts of esters of fatty acids and carnitine; alkyl sulfate salts; fatty acid salts; docusate sodium; acyllactylates; mono- and diacetylated esters of tartaric acid and mono- and diglycerides; succinylated mono- and diglycerides; esters of citric acid and mono- and diglycerides; and mixtures thereof.

Among the aforementioned group, ionic surfactants include, by way of example: lecithins, lysolecithin, phospholipids, lysophospholipids and their derivatives; salts of esters of fatty acids and carnitine; alkyl sulfate salts; fatty acid salts; docusate sodium; acyllactylates; mono- and diacetylated esters of tartaric acid and mono- and diglycerides; succinylated mono- and diglycerides; esters of citric acid and mono- and diglycerides; and mixtures thereof.

Ionic surfactants can be ionized forms of lecithin, lysolecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidyl ethanolamine-l-phosphonyl-phosphonylamine-lysophosphate lysophosphate lysophosphate lysophosphate lysophosphate lysophosphate acids, stearoyl-2-lactylate, stearoyl lactylate, succinylated monoglycerides,

mono / diacetylated esters of tartaric acid and mono / diglycerides, esters of citric acid and mono / diglycerides, cholylsarcosine, caproate, caprylate, caprate, laurate, myristate, palmitate, oleate, ricinoleate, linoleate, linolenate, stearalate, teuris lauryl docusate, lauroyl-carnitines, palmitoyl-carnitines, myristoyl-carnitines and their salts and mixtures.

Hydrophilic nonionic surfactants may include, but are not limited to, alkyl glucosides; alkyl maltosides; alkylthioglucosides; lauryl macrogolglycerides; polyoxyalkylene alkyl ethers such as polyethylene glycol alkyl ethers; polyoxyalkylene alkyl phenols such as polyethylene glycol alkyl phenols; polyoxyalkylene fatty acid and alkyl phenol esters such as fatty acid and polyethylene glycol monoesters and fatty acid and polyethylene glycol diesters; fatty acid esters of polyethylene glycol glycerol; fatty acid esters of polyglycerol; fatty acid esters of polyoxyalkylene sorbitan, such as fatty acid esters of polyethylene glycol sorbitan; hydrophilic products of transesterification of a polyol with at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols; polyoxyethylene sterols, their derivatives and analogues; polyoxyethylated vitamins and their derivatives; block copolymers of polyoxyethylene and polyoxypropylene and mixtures thereof; fatty acid esters of polyethylene glycol sorbitan; and hydrophilic transesterification products of a polyol with at least one member of the group consisting of triglycerides, vegetable oils, and hydrogenated vegetable oils. The polyol may be glycerol, ethylene glycol, polyethylene glycol, sorbitol, propylene glycol, pentaerythritol or saccharide.

Other hydrophilic non-ionic surfactants include, but are not limited to, PEG-10 Laurate, PEG-12 Laurate, PEG-20 Laurate, PEG-32 Laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32 distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25 glyceryl trioleate, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30 glyceryl laurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate, PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryl pearl, oil 50 hydr genetized castor oil, PEG-40 castor oil, PEG-35 castor oil, PEG-60 castor oil, PEG-40 hydrogenated castor oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6 caprate / caprylate glycerides PEG-8 caprate / caprylate glycerides, polyglyceryl-10-laurate, PEG-30 cholesterol, PEG-25 phytosterol, PEG-30 soy sterol, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitan laurate, polysorbate 20, Polysorbate 80, POE-9 lauryl ether, POE-23 lauryl ether, POE-10 oleyl ether, POE-20 pro toy oleyl ether, POE-20 stearyl ether, tocopheryl-PEG-100-succinate, PEG-24-cholesterol, polyglyceryl-10-oleate, Tween 40, Tween 60, sucrose monostearate, sucrose monolaurate, sucrose monopalmitate, PEG 10- series 100 nonylphenols, a series of PEG 15-100 octylphenols and poloxamers.

Suitable lipophilic surfactants include, by way of example only: fatty alcohols; fatty acid esters of glycerol; acetylated fatty acid esters of glycerol; esters of fatty acids and lower alcohols; fatty acid esters of propylene glycol; fatty acid esters of sorbitan; fatty acid esters of polyethylene glycol sorbitan; sterols and derivatives of sterols; polyoxyethylene sterols and sterol derivatives; alkyl ethers of polyethylene glycols; sugar esters; sugar ethers; derivatives of lactic acid and mono- and diglycerides; hydrophobic polyester transesterification products with at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols; oil soluble vitamins / vitamin derivatives; and mixtures thereof. Among this group, preferred lipophilic surfactants include fatty acid esters of glycerol, fatty acid esters of propylene glycol and mixtures thereof, or they are hydrophobic products of transesterification of a polyol with at least one member of the group consisting of vegetable oils, hydrogenated vegetable oils and triglycerides.

In one embodiment, the composition may include a solubilizer to provide good solubilization and / or dissolution of the compounds of the present invention and to minimize precipitation of the compounds of the present invention. This may be especially important for compositions intended for non-oral administration, for example, compositions for injection. The solubilizer can also be added to increase the solubility of the hydrophilic drug and / or other components, such as surfactants, or to maintain the composition in the form of a stable or homogeneous solution or dispersion.

Examples of suitable solubilizers include, but are not limited to, the following: alcohols and polyols such as ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediols and their isomers, glycerol, pentaerythritol, sorbitol, mannitol, transcutol, dimethyl isorb polyethylene glycol, polypropylene glycol, polyvinyl alcohol, hydroxypropyl methyl cellulose and other cellulose derivatives, cyclodextrins and cyclodextrins derivatives; ethers of polyethylene glycols having an average molecular weight of from about 200 to about 6000, such as ether of PEG and tetrahydrofurfuryl alcohol (glycofurol) or methoxy-PEG; amides and other nitrogen-containing compounds such as 2-pyrrolidone, 2-piperidone, epsilon-caprolactam, N-alkylpyrrolidone, N-hydroxyalkylpyrrolidone, N-alkylpiperidone, N-alkylcaprolactam, dimethylacetamide and polyvinylpyrrolidone; esters such as ethyl propionate, tributyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, triethyl citrate, ethyl oleate, ethyl caprylate, ethyl butyrate, triacetin, propylene glycol monoacetate, δ-isomerol, isomerolone and other solubilizers known in the art, such as dimethylacetamide, dimethyl isosorbide, N-methylpyrrolidones, monooctanoin, diethylene glycol monoethyl ether and water.

In addition, mixtures of solubilizers can be used. Examples include, but are not limited to, triacetin, triethyl citrate, ethyl oleate, ethyl caprylate, dimethylacetamide, N-methylpyrrolidone, N-hydroxyethyl pyrrolidone, polyvinylpyrrolidone, hydroxypropylmethyl cellulose, hydroxypropyl cyclodextrins, ethanol glycol, ethanol, glycol 200, glycol, ethanol, glycol 200, and dimethyl isosorbide. Particularly preferred solubilizers include sorbitol, glycerin, triacetin, ethyl alcohol, PEG-400, glycofurol and propylene glycol.

The amount of solubilizer that can be included is not particularly limited. The amount of a given solubilizer can be limited by a biologically acceptable amount, which can be easily determined by a person skilled in the art. In some cases, it may be advantageous to include an amount of solubilizers that is much higher than biologically acceptable amounts, for example, to maximize the concentration of the drug, while the excess of solubilizer is removed before the composition is administered to the patient using traditional techniques such as distillation or evaporation. So, if present, the solubilizer can be in a mass ratio of 10%, 25%, 50%, 100%, or up to about 200% by weight inclusive, based on the combined weight of the drug and other excipients. If necessary, very small amounts of solubilizer, such as 5%, 2%, 1% or even less, can also be used. Typically, the solubilizer may be present in an amount of from about 1% to about 100%, more typically from about 5% to about 25% by weight.

In addition, the composition may include one or more than one of the pharmaceutically acceptable excipients and excipients. Such excipients and excipients include, but are not limited to, tackifiers, anti-foaming agents, buffering agents, polymers, antioxidants, preservatives, chelating agents, viscosity modifying agents, tonicity modifying agents, flavoring agents, coloring agents, perfumes, mufflers, suspending agents agents, binders, fillers, emollients, lubricants and mixtures thereof.

In addition, an acid or base may be included in the composition to facilitate manufacturing techniques, to enhance stability, or for other reasons. Examples of pharmaceutically acceptable bases include amino acids, amino acid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide, sodium hydrogen carbonate, aluminum hydroxide, calcium carbonate, magnesium hydroxide, magnesium aluminum silicate, synthetic aluminum silicate, synthetic hydrocalcite, magnesium aluminum hydroxide, diisopropylethylamine, ethanolamine, ethylene diamine , triethanolamine, triethylamine, triisopropanolamine, trimethylamine, tris (hydroxymethyl) aminomethane (TRIS) and the like. Bases which are salts of a pharmaceutically acceptable acid such as acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carboxylic acid, citric acid, fatty acids are also suitable. , formic acid, fumaric acid, gluconic acid, hydroquinonsulfonic acid, isoascorbic acid, lactic acid, maleic acid, oxalic acid, lara-bromophenyl sulfonic I acid, propionic acid, l-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid and the like. Polyprotic acid salts such as sodium phosphate, disodium hydrogen phosphate and sodium dihydrogen phosphate can also be used. If the base is a salt, then the cation may be any convenient and pharmaceutically acceptable cation, such as ammonium, cations of alkali metals, alkaline earth metals and the like. An example may include, but is not limited to, sodium, potassium, lithium, magnesium, calcium, and ammonium.

Suitable acids are pharmaceutically acceptable organic or inorganic acids. Examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, boric acid, phosphoric acid and the like. Examples of suitable organic acids include acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acids, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carboxylic acid, citric acid, fatty acids, formic acid, fumaric acid, acid, hydroquinonsulfonic acid, isoascorbic acid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid, para-bromophenyl sulfonic acid, propionic acid acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid and the like.

Pharmaceutical compositions for injection. In some embodiments of the present invention, there is provided a pharmaceutical composition for injection comprising a compound of the present invention and a pharmaceutical excipient suitable for injection. Components and amounts of agents in such compositions are as described herein.

Forms into which the new compositions of the present invention for injection can be included include aqueous or oily suspensions or emulsions with sesame oil, corn oil, cottonseed oil or peanut oil, as well as elixirs, mannitol, dextrose or a sterile aqueous solution, and similar pharmaceutical diluents.

Aqueous solutions in saline are also traditionally used for injection. Ethanol, glycerin, propylene glycol, liquid polyethylene glycol and the like (and their suitable mixtures), cyclodextrin derivatives and vegetable oils may also be used. Proper fluidity can be maintained, for example, by using a coating agent, such as lecithin, to maintain the desired particle size in the case of dispersion and by using surfactants. Prevention of the action of microorganisms can be carried out by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.

Sterile injectable solutions are prepared by introducing the compound of the present invention in the required amount into an appropriate solvent along with various other ingredients, which are listed above, as necessary, followed by sterilization by filtration. Typically, dispersions are prepared by introducing various sterile active ingredients into a sterile diluent that contains a basic dispersing medium and the necessary other ingredients from those listed above. In the case of sterile powders for the preparation of sterile injectable solutions, some desired manufacturing methods are vacuum drying and lyophilization drying methods, which allow the preparation of the active ingredient powder plus any additional desired ingredient from a solution subjected to pre-sterilization by filtration.

Pharmaceutical compositions for topical (eg, transdermal) delivery. In some embodiments of the present invention, there is provided a pharmaceutical composition for transdermal delivery comprising a compound of the present invention and a pharmaceutical excipient suitable for transdermal delivery.

The compositions of the present invention can be made in the form of preparations in solid, semi-solid or liquid forms suitable for local or local administration, such as gels, water-soluble jellies, creams, lotions, suspensions, foams, powders, suspensions, ointments, solutions, oils, pastes, suppositories, sprays, emulsions, saline solutions, solutions based on dimethyl sulfoxide (DMSO). As a rule, the use of carriers having higher density values makes it possible to obtain a region with prolonged action of the active ingredients. In contrast, a composition in solution may provide a more rapid action of the active ingredient on a selected area.

The pharmaceutical compositions may also contain suitable carriers or excipients in the solid or gel phase, which are compounds that increase penetration or facilitate the delivery of therapeutic agents through the permeability barrier defined by the stratum corneum of the skin. There are many such penetration enhancing molecules known to those skilled in the art of topical compositions. Examples of such carriers and excipients include, but are not limited to, humectants (e.g., urea), glycols (e.g., propylene glycol), alcohols (e.g., ethanol), fatty acids (e.g., oleic acid), surfactants (e.g., isopropyl myristate and sodium lauryl sulfate), pyrrolidones, glycerol monolaurate, sulfoxides, terpenes (e.g. menthol), amines, amides, alkanes, alkanols, water, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin and polymers such as polyethylene glycol .

In another typical composition intended for use in the methods of the present invention, transdermal delivery devices (“patches”) are used. Such transdermal patches can be used to provide continuous or intermittent infusion of the compounds of the present invention in controlled amounts, either in the presence or absence of another agent.

The design and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. See, for example, US Patent Nos. 5023252, 4992445 and 5001139. Such patches can be designed for continuous, pulsating delivery or delivery of pharmaceutical agents as needed.

Pharmaceutical compositions for inhalation. Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. Preferably, the compositions are administered by the oral or nasal route through the respiratory tract to provide local or systemic effects. Compositions, preferably in pharmaceutically acceptable solvents, can be nebulized using inert gases. Spray solutions can be inhaled directly from the spray device or the spray device can be attached to a respirator, or using a breathing apparatus operating under intermittent positive pressure. Compositions in the form of a solution, suspension or powder can be administered, preferably orally or nasally, from devices by which the composition is delivered in an appropriate manner.

Other pharmaceutical compositions. Pharmaceutical compositions can also be made from the compositions set forth herein and one or more pharmaceutically acceptable excipients suitable for sublingual, buccal, rectal, intraosseous, intraocular, intranasal, epidural or intraspinal administration. The manufacture of such pharmaceutical compositions is well known in the art. See, for example, Anderson, Philip O .; Knoben, James E .; Troutman, William G, eds., Handbook of Clinical Drug Data, tenth edition, McGraw-Hill, 2002; Pratt and Taylor, eds., Principles of Drug Action, third edition, Churchill Livingston, New York, 1990; Katzung, ed., Basic and Clinical Pharmacology, ninth edition, McGraw Hill, 20037ybg; Goodman and Gilman, eds., The Pharmacological Basis of Therapeutics, tenth edition, McGraw Hill, 2001; Remingtons Pharmaceutical Sciences, 20th Edition, Lippincott Williams & Wilkins., 2000; Martindale, The Extra Pharmacopoeia, thirty-second edition (The Pharmaceutical Press, London, 1999); all of them are included in this description in its entirety by reference.

The administration of the compounds or pharmaceutical composition of the present invention can be carried out by any method that delivers the compounds to the site of action. These methods include oral methods, intraduodenal methods, parenteral injection (including intravenous, intraarterial, subcutaneous, intramuscular, intravascular, intraperitoneal or infusion), topical (e.g. transdermal applications), rectal administration, via local delivery using a catheter or stent or by inhalation . The compounds can also be administered orally or intrathecally.

The amount of compound administered will depend on the mammal being treated, the severity of the disorder or condition, the rate of administration, the pharmacokinetics of the compound, and the choice of the attending physician. However, an effective dosage is in the range of from about 0.001 to about 100 mg per kg of body weight per day, preferably from about 1 to about 35 mg / kg / day in single or divided doses. For a person weighing 70 kg, this amount will be from about 0.05 to 7 g / day, preferably from about 0.05 to about 2.5 g / day. In some cases, dosage levels below the lower limit of the above range may be more adequate, while in other cases substantially higher doses may be used, which will not cause any dangerous side effect, for example, by splitting such large doses into several small doses for administration during the day.

In some embodiments, the compound of the invention is administered in a single dose. Typically, such administration will be by injection, for example, by intravenous injection, in order to rapidly administer the agent. However, other methods that may be appropriate may be used. A single dose of a compound of the invention can also be used to treat an acute condition.

In some embodiments, the compound of the invention is administered in multiple doses. The introduction can be performed approximately once, twice, three times, four times, five times, six times or more than six times a day. Administration can be performed approximately once a month, once every two weeks, once a week, or once every subsequent day. In another embodiment, the compound of the invention and another agent are administered together from about once a day to about 6 times a day. In another embodiment, the administration of the compound of the invention and the agent is continued for less than about 7 days. In yet another embodiment, administration is continued for more than about 6, 10, 14, 28 days, two months, six months, or one year. In some cases, they are administered continuously and maintained for as long as necessary.

The administration of the agents of the invention may continue as long as necessary. In some embodiments, the agent of the invention is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, the agent of the invention is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, the agent of the invention is administered continuously, for example, for treating chronic effects.

An effective amount of a compound of the invention can be administered either in a single dose or in multiple doses by any of the acceptable routes of administration of agents having similar uses, including rectal, buccal, intranasal and transdermal methods, by intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly , subcutaneously, orally, topically, or as an inhalant.

The compositions of the invention can also be delivered using an impregnated or coated device, such as a stent, for example, or a cylindrical-shaped polymer material introduced into the artery. Such a route of administration may help, for example, to prevent or reduce restenosis after procedures such as balloon angioplasty. Without being bound by theory, it is believed that the compounds of the invention can slow down or inhibit the migration and proliferation of smooth muscle cells in the artery wall, which contribute to restenosis. The compound of the invention may be administered, for example, by local delivery from stent strata, from a stent transplant, from transplants, or from a coating or sheath of a stent. In some embodiments, the compound of the invention is mixed with a matrix. Such a matrix may be a polymer matrix and may serve to bind the compound to the stent. Polymer matrices suitable for this use include, for example, polyesters or lactone-based polyesters such as polylactide, polycaprolactone glycolide, polyorthoesters, polyanhydrides, polyamino acids, polysaccharides, polyphosphazenes, poly - [(ether) copolymers ( ether)] (for example, PEO-PLLA (copolymer based on polyethylene oxide and polylactic acid)); polydimethylsilokeane, poly (ethylene-vinyl acetate), acrylate-based polymers or copolymers (e.g. polyhydroxyethyl methyl methacrylate, polyvinyl pyrrolidinone), fluorinated polymers such as polytetrafluoroethylene and cellulose esters. Suitable matrices may be non-degradable or may decompose over time, releasing the compound or compounds. The compounds of the invention can be applied to the surface of the stent in various ways, such as immersion / centrifugal coating, spray coating, immersion coating and / or coating with a brush device. The compound can be applied in a solvent and the solvent can be left to evaporate, resulting in a layer of the compound on the stent. Alternatively, the connection can be localized in the stent or graft body, for example in microchannels or micropores. After implantation, the compound diffuses from the stent body to contact the artery wall. Such stents can be made by immersing a stent made to contain such micropores or microchannels in a solution of the compound of the invention in a suitable solvent, followed by evaporation of the solvent. Excess drug on the surface of the stent can be removed by additional short-term washing with a solvent. In further other embodiments, the compounds of the invention may be covalently linked to a stent or graft. You can use the covalent linker, which is destroyed in vivo, leading to the release of the compounds of the invention. For this purpose, any biologically labile bond, such as, for example, ester, amide or anhydride bonds, can be used. The compounds of the invention may be additionally administered intravascularly from a balloon used in the process of angioplasty. To reduce restenosis, extravascular administration of the compounds can also be accomplished by introducing the compositions of the invention through the pericardium or adventitia membrane.

A variety of stent devices that can be used as described are disclosed, for example, in the following references, all of which are hereby incorporated by reference: US Pat. No. 5,451,233; US patent No. 5040548; US patent No. 5061273; U.S. Patent No. 5,496,346; U.S. Patent No. 5,292,331; U.S. Patent No. 5,674,278; U.S. Patent No. 3657744; U.S. Patent No. 4,739,762; U.S. Patent No. 5,195,984; U.S. Patent No. 5,292,331; U.S. Patent No. 5,674,278; U.S. Patent No. 5,879,382; US patent No. 6344053.

The compounds of the invention may be administered in dosage. It is known in the art that due to interpersonal variability in the pharmacokinetics of compounds, individualization of the administration regimen is a necessary feature for optimal therapy. The dosage regimen for the compound of the invention can be determined based on routine experimentation in light of the present description.

In the case where the compound of the invention is incorporated into a composition that contains one or more agents and this agent has a shorter half-life than the compound of the invention, the unit dosage forms of the agent and the compounds of the invention can be adjusted accordingly.

The pharmaceutical composition of the invention may, for example, be in a form suitable for oral administration in the form of a tablet, capsule, pill, powder, continuous release composition, solution, suspension, for parenteral injection in the form of a sterile solution, suspension or emulsion, for topical administration as an ointment or cream, or for rectal administration as a suppository. Such a pharmaceutical composition may be in unit dosage forms suitable for single administration of exact dosages. The pharmaceutical composition will include a conventional pharmaceutical carrier or excipient and a compound of the invention as an active ingredient. In addition to this, it may include other medical or pharmaceutical agents, carriers, adjuvants, etc.

Typical forms for parenteral administration include solutions or suspensions of the active compound in sterile aqueous solutions, for example, aqueous propylene glycol or dextrose solutions. Such dosage forms may be suitably buffered, if necessary.

The activity of the compounds of the present invention can be determined according to the following procedure, as well as the procedure described below in the Examples section. The kinase activity is evaluated by measuring, in the presence of a kinase, the incorporation of γ- ³³ P-phosphate from γ- ³³ P-ATP into an N-terminal His-tag substrate that is expressed in E. coli and which is purified by conventional methods. This assay is performed in a 96-well polypropylene plate. The incubated mixture (100 μl) contains 25 mM Hepes (N-2-hydroxyethyl-piperazine-N-2-ethanesulfonic acid), pH 7.4, 10 mM MgCl ₂ , 5 mM β-glycerophosphate, 100 μM Na-orthovanadate, 5 mM DTT (dithiothreitol), 5 nM kinase and 1 μM substrate. Inhibitors are suspended in DMSO and all reactions, including controls, are performed at a final concentration of DMSO of 1%. Reactions are initiated by the addition of 10 μM ATP (at a dose of 0.5 μCi γ- ³³ P-ATP / well) and incubated at ambient temperature for 45 minutes. An equal volume of 25% TCA (trichloroacetic acid) is added to stop the reaction and precipitate the proteins. The precipitated proteins are captured on filter plates with a fiberglass B filter and the excess labeled ATP is washed using a Tomtec MACH III harvester. The plates were allowed to air dry, after which 30 μl Packard Microscint 20 was added to each well and the plates were counted using Packard TopCount.

Kits are also provided according to the invention. Kits include a compound or compounds of the present invention, as set forth herein, in suitable packaging, and a written document, which may include instructions for use, discussion of a clinical trial, list of side effects, and the like. Such kits may also include information such as links from the scientific literature, materials related to the instructions for use of the drug, the results of clinical trials and / or a brief report on them and the like, which indicate or establish the activities and / or advantages of this composition and / or in which dosages, routes of administration, side effects, drug interactions, or other information that is useful to health care providers and specialists are described. Such information may be based on the results of various studies, for example studies using experimental animals participating in in vivo models and studies based on clinical trials in humans. This kit may further comprise another agent. In some embodiments, the compound of the present invention and the agent are provided in a kit as separate compositions in separate containers. In some embodiments, the compound of the present invention and the agent are provided in a kit as a single composition in a container. Suitable packaging and additional products for use (for example, a measuring cup for liquid preparations, foil wrapping to minimize exposure to air, and the like) are known in the art and can be included in a kit. The kits described in this application can be offered, put up for sale and / or advertised to institutions and specialists providing medical services, including doctors, nurses, pharmacists, officials responsible for compiling the formulations of drugs, and the like. In addition, kits, in some embodiments, can be sold directly to the consumer.

Ways

In addition, the invention provides methods of using the compounds or pharmaceutical compositions of the present invention for the treatment of disease states, including, but not limited to, diseases associated with the malfunctioning of one or more types of PI3 kinase. A detailed description of the conditions and disorders mediated by p110δ-kinase activity is given in the application of Sadu et al., WO 01/81346, which is incorporated herein in its entirety by reference for all purposes.

The treatment methods provided herein include administering to a subject a therapeutically effective amount of a compound of the invention. In one embodiment of the present invention, there is provided a method of treating an inflammatory disorder, including autoimmune diseases, in a mammal. The method comprises administering to said mammal a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof. Examples of autoimmune diseases include, but are not limited to, acute disseminating encephalomyelitis (ADEM), Addison’s disease, antiphospholipid syndrome (APS), hypoplastic anemia, autoimmune hepatitis, gluten enteropathy, Crohn’s disease, diabetes mellitus, type 1 diabetes, Good's syndrome , Guillain-Barré Syndrome (GBS), Hashimoto’s disease, lupus erythematosus, multiple sclerosis, severe pseudoparalytic myasthenia gravis, Opsoclonus myoclonus syndrome (OMS), retrobulbar neuritis, Horde thyroiditis, pemphigus, polyarthritis, primary biliary cirrhosis, psoriasis, rheumatoid arthritis, Reiter’s syndrome, Takayasu arteritis, temporal arteritis (also known as “giant cell arteritis”), autoimmune hemolytic anemia with thermal agglutinin syndrome, Wegener's granulomatosis, universal alopecia, Chagas disease, chronic fatigue syndrome, , endometriosis, purulent hydradenitis, interstitial cystitis, neuromyotonia, sarcoidosis, scleroderma, ulcerative colitis, vitiligo and vulvodynia. Other disorders include bone resorption disorders and thrombosis.

In some embodiments, a method of treating inflammatory or autoimmune diseases comprises administering to a subject (eg, a mammal) a therapeutically effective amount of one or more compounds of the present invention that selectively inhibit PI3K-δ and / or PI3K-γ compared to all other type I PI3 kinases . Such selective inhibition of PI3K-δ and / or PI3K-γ may be advantageous in the treatment of any of the diseases or conditions described herein. For example, selective inhibition of PI3K-δ can inhibit inflammatory responses associated with inflammatory diseases, autoimmune disease, or diseases associated with an undesirable immune response, including, but not limited to, asthma, emphysema, allergies, dermatitis, rheumatoid arthritis, psoriasis, lupus erythematosus or graft versus host reaction. In addition, selective inhibition of PI3K-δ can provide a decrease in the inflammatory or undesirable immune response without a concomitant decrease in the ability to attenuate bacterial, viral and / or fungal infections. Selective inhibition of both PI3K-δ and PI3K-γ may have a greater advantage in inhibiting the inflammatory response in a subject than would be obtained using inhibitors that selectively inhibit only PI3K-δ or PI3K-γ. In one aspect, one or more of the methods of the invention is effective in reducing the production of antigen-specific antibodies in vivo by about 2 times, 3 times, 4 times, 5 times, 7.5 times, 10 times, 25 times, 50 times, 100 times, 250 times, 500 times, 750 times, or about 1000 times or more. In another aspect, one or more of the methods of the invention is effective in reducing the production of antigen-specific IgG3 and / or IgGM in vivo by about 2 times, 3 times, 4 times, 5 times, 7.5 times, 10 times, 25 times, 50 times, 100 times, 250 times, 500 times, 750 times, or about 1000 times or more.

In one aspect, one or more of the methods of the invention is effective in reducing the intensity of symptoms associated with rheumatoid arthritis, including, but not limited to, reducing joint swelling, reducing serum anti-collagen antibodies and / or reducing joint pathology, such as bone resorption, cartilage damage, pannus and / or inflammation. In another aspect, the methods of the invention are effective in reducing ankle inflammation by at least about 2%, 5%, 10%, 15%, 20%, 25%, 30%, 50%, 60%, or about 75% -90%. In another aspect, the methods of the invention are effective in reducing knee joint inflammation by at least about 2%, 5%, 10%, 15%, 20%, 25%, 30%, 50%, 60%, or about 75% -90% or more. In yet another aspect, the methods of the invention are effective in decreasing serum levels of antibodies against type II collagen by at least about 10%, 12%, 15%, 20%, 24%, 25%, 30%, 35 %, 50%, 60%, 75%, 80%, 86%, 87%, or about 90% or more. In another aspect, the methods of the invention are effective in decreasing the histopathological scale for the ankle joint by about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 75% , 80%, 90% or more. In yet another aspect, the methods of the invention are effective in reducing histopathological score for the knee joint by about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 75%, 80%, 90% or more.

In other embodiments of the present invention, methods of using the compounds or pharmaceutical compositions for the treatment of respiratory diseases, including, but not limited to, diseases affecting the lung lobes, pleural cavity, bronchioles, trachea, upper respiratory tract or nerves and muscles involved in the act of breathing, are provided. For example, methods for treating obstructive pulmonary disease are provided. Chronic obstructive pulmonary disease (COPD) is a generic term for a group of respiratory tract diseases that are characterized by obstruction or airway restriction. The conditions included in this generalizing term are: chronic bronchitis, emphysema and bronchiectasis.

In another embodiment, the compounds described herein are used to treat asthma. In addition, the compounds or pharmaceutical compositions described herein can be used to treat endotoxemia and sepsis. In one embodiment, the compounds or pharmaceutical compositions set forth herein are used to treat rheumatoid arthritis (RA). In yet another embodiment, the compounds or pharmaceutical compositions described herein are used to treat contact or atopic dermatitis. Contact dermatitis includes irritating dermatitis, phototoxic dermatitis, allergic dermatitis, photoallergic dermatitis, contact urticaria, contact type systemic dermatitis and the like. Irritating dermatitis can occur when too much substance is applied to the skin or when the skin is sensitive to a particular substance. Atopic dermatitis, sometimes called eczema, is a type of dermatitis - atopic skin disease.

The invention also relates to a method for treating a hyperproliferative disorder in a mammal, comprising administering to said mammal a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof. In some embodiments, the method relates to the treatment of cancer, such as acute myeloid leukemia, cancer of the thymus, brain, lung, squamous cell, skin, eye, retinoblastoma, intraocular melanoma, cancer of the oral cavity and oropharynx, bladder, gastrointestinal tract, stomach, pancreas, bladder, breast, cervix, head, neck, renal sphere, kidney, liver, ovaries, prostate, colorectal cancer, cancer of the esophagus, testis, gynecological sphere, thyroid gland, central nervous system, PNS, AIDS-related (e.g., lymphoma and Kaloshi sarcoma) or virus-induced cancer. In some embodiments, said method relates to the treatment of benign hyperproliferative disorder, such as benign skin hyperplasia (e.g., psoriasis), restenosis, or benign hyperproliferative disorder of the prostate (e.g., benign prostatic hypertrophy (BPH)).

The invention also relates to a method for treating a disease associated with vasculogenesis or angiogenesis in a mammal, comprising administering to said mammal a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof. In some embodiments, the method relates to the treatment of a disease selected from the group consisting of tumor angiogenesis, a chronic inflammatory disease such as rheumatoid arthritis, atherosclerosis, inflammatory bowel disease, skin diseases such as psoriasis, eczema and scleroderma, diabetes, diabetic retinopathy, retinopathy of premature, age-related macular degeneration, hemangioma, glioma, melanoma, Kaposi’s sarcoma and cancer of the ovaries, breast, lung, pancreas, and Yelnia prostate, colon and epidermoid cancer.

Patients who may be treated with the compounds of the present invention or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative of these compounds in accordance with the methods of this invention include, for example, patients who have been diagnosed with psoriasis; restenosis; atherosclerosis; BPH; breast cancer, such as ductal carcinoma of the breast duct tissue, types of medullary cancer, types of colloidal cancer, tubular carcinomas, and inflammatory breast cancer; ovarian cancer, including ovarian epithelial tumors such as adenocarcinoma in the ovary and adenocarcinoma that migrates from the ovary to the abdominal cavity; uterine cancer; cervical cancer, such as adenocarcinoma in the cervical epithelium, including squamous cell carcinoma and adenocarcinomas; prostate cancer, for example, prostate cancer selected from the following: adenocarcinomas or adenocarcinomas that migrate to the bone; pancreatic cancer, such as epithelioid carcinoma in the tissue of the pancreatic ducts and adenocarcinoma in the pancreatic duct; bladder cancer, such as transitional cell carcinoma in the bladder, urothelial carcinomas (transitional cell carcinomas), tumors in urothelial cells lining the bladder, squamous cell carcinoma, adenocarcinoma and small cell carcinoma; leukemia, such as acute myeloid leukemia (AML), acute lymphocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, myelodysplasia, myeloproliferative disorders, acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), chronic myelogen lymphocytic leukemia (CML), (CLL), multiple myeloma (MM) and myelodysplastic syndrome (MDS); bone cancer lung cancer, such as non-small cell lung cancer (NSCLC), which is divided into squamous cell carcinoma, adenocarcinomas and large cell non-differentiable carcinomas, and small cell lung cancer; skin cancer, such as basal cell carcinoma, melanoma, squamous cell carcinoma and actinic keratosis, which is a skin condition that sometimes develops into squamous cell carcinoma; ophthalmic retinoblastoma; cutaneous or intraocular melanoma (eye melanoma); primary liver cancer (cancer that begins in the liver); kidney cancer thyroid cancer, such as papillary, follicular, medullary and anaplastic; AIDS-associated lymphoma, such as diffuse B-large cell lymphoma, B-cell immunoblastic lymphoma and small cell lymphoma from undigested cells; Kaposi's sarcoma; types of cancer induced by viruses, including cancer induced by hepatitis B virus (HBV), hepatitis C virus (HCV), and hepatocellular carcinoma; cancer induced by human T-type Lymphotropic virus type 1 (HTLV-1), and adult T-cell leukemia / lymphoma; and human papilloma virus (HPV) induced cancer and cervical cancer; types of cancer of the central nervous system (CNS), such as a primary brain tumor that includes gliomas (astrocytoma, anaplastic astrocytoma or glioblastoma multiforme), oligodendroglioma, ependymoma, meningioma, lymphoma, schwannomas and medulloblastoma; types of cancer of the peripheral nervous system (PNS), such as acoustic neuroma and malignant neoplasms of the peripheral nerves (MPNST), including neurofibromas and schwannomas, malignant fibrotic cytoma, malignant fibrotic histiocytoma, malignant meningioma, malignant mesomelioma; cancer of the oral cavity and oropharynx, such as cancer of the submucosal part of the pharynx, cancer of the larynx, nasopharyngeal cancer and oropharyngeal cancer; gastric cancer, such as lymphomas, stromal tumors and carcinoid tumors of the gastrointestinal tract; testicular cancer, such as embryonic cell tumors (GCT), which include seminomas and non-seminomic tumors, and gonadal stromal tumors, which include tumors from Leydig cells and tumors from Sertoli cells; thymus cancer, such as thymomas, thymic carcinomas, Hodgkin's disease, non-Hodgkin lymphomas, carcinoids or carcinoid tumors; rectal cancer; and colon cancer.

The invention also relates to a method for treating diabetes in a mammal, comprising administering to said mammal a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof.

In addition, the compounds described in this application can be used to treat acne.

In addition, the compounds described in this application can be used to treat arteriosclerosis, including atherosclerosis. Arteriosclerosis is a general term that describes any sclerosis of the medium or large arteries. Atherosclerosis is arterial sclerosis, mainly due to atherosclerotic plaque.

Other compounds set forth in this application can be used to treat glomerulonephritis. Glomerulonephritis is a primary or secondary autoimmune kidney disease characterized by glomerular inflammation. It may be asymptomatic or occur with hematuria and / or proteinuria. There are many generally recognized types, which are divided into acute, subacute or chronic glomerulonephritis. The causes are infectious (bacterial, viral or parasitic pathogens), autoimmune or paraneoplastic.

In addition, the compounds described in this application can be used to treat bursitis, lupus, acute disseminating encephalomyelitis (ADEM), Addison’s disease, antiphospholipid syndrome (APS), hypoplastic anemia, autoimmune hepatitis, gluten enteropathy, Crohn’s disease, (Type 1), Goodpast syndrome, Graves disease, Guillain-Barré syndrome (GBS), Hashimoto’s disease, inflammatory bowel disease, lupus erythematosus, severe pseudoparalytic myasthenia gravis, Opsoclonus myoclonus syndrome (OMS), retrobulb neuritis, thyroiditis, Horde, osteoarthritis, uveoretinitis, pemphigus, polyarthritis, primary biliary cirrhosis, Reiter's syndrome, Takayasu arteritis, temporal arteritis, autoimmune hemolytic anemia with heat agglutinins syndrome, Wegener's granulomatosis, universal dysonopathy, alopecia areata , endometriosis, purulent hydradenitis, interstitial cystitis, neuromyotonia, sarcoidosis, scleroderma, ulcerative colitis, vitiligo, vulvodynia, appendicitis, arteritis, and trita, blepharitis, bronchiolitis, bronchitis, cervicitis, cholangitis, cholecystitis, chorioamnionitis, colitis, conjunctivitis, cystitis, dacryoadenitis, dermatomyositis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, gastritis, gastrititis, epididitis, gastritis, gastrititis, epididitis, gastritis, gastrititis hydradenitis, ileitis, iritis, laryngitis, mastitis, meningitis, myelitis, myocarditis, myositis, nephritis, omphalitis, oophoritis, orchitis, osteitis, otitis media, pancreatitis, mumps, pericarditis, peritonitis, pharyngitis, pleurisy, phlebitis, prostatitis, phlebitis, and, pyelonephritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis, tendonitis, tonsillitis, uveitis, vaginitis, vulvitis, or vasculitis.

The invention also relates to a method for treating cardiovascular disease in a mammal, comprising administering to said mammal a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof. Examples of cardiovascular conditions include, but are not limited to, atherosclerosis, restenosis, vascular occlusion, and obstructive carotid artery disease.

In another aspect of the present invention, methods for dysfunction of white blood cells or dysfunction of osteoclasts are provided. The method involves bringing leukocytes or osteoclasts into contact with a function-disrupting amount of a compound of the invention.

In another aspect of the present invention, methods are provided for treating an ocular disease by administering one or more than one compound or one or more of the pharmaceutical compositions of the invention into the eye of a subject.

In addition, methods of administering the compounds of the present invention by eye drops, intraocular injection, intravitreal injection, topically or by using a drug eluting device, microcapsules, implant or microjet device are provided. In some cases, the compounds of the present invention are administered together with a carrier or excipient that improves the penetration of the compound into the eye, such as an oil and water emulsion with colloidal particles having an oil core surrounded by a film at the phase boundary.

In some cases, colloidal particles include at least one cationic agent and at least one non-ionic surfactant, such as poloxamer, tyloxapol, polysorbate, a polyoxyethylene and castor oil derivative, a sorbitan ester or polyoxy stearate. In some cases, the cationic agent is an alkylamine, a tertiary alkylamine, a quaternary ammonium compound, a cationic lipid, an amino alcohol, a biguanide salt, a cationic compound, or a mixture thereof. In some instances, the cationic agent is a biguanide salt such as chlorhexidine, polyaminopropyl biguanide, phenformin, alkyl biguanide, or a mixture thereof. In some cases, the quaternary ammonium compound is benzalkonium halide, lauralconium halide, cetrimide, hexadecyl trimethylammonium halide, tetradecyl trimethyl ammonium halide, dodecyl trimethyl ammonium halide, benzene halide, cetethyldimonium halide (cetethyldimonium halide), cetyl pyridinium halide, benzododecinium halide, chlorallyl-methenamine-halide, myristylalkonium halide, stearalconium halide or a mixture of two or olee than two of them. In some cases, the cationic agent is benzalkonium chloride, lauralconium chloride, benzododecinium bromide, benzethonium chloride, hexadecyl trimethyl ammonium bromide, tetradecyl trimethyl ammonium bromide, dodecyl trimethyl ammonium bromide, or a mixture of two or more thereof. In some cases, the oil phase is mineral oil and light mineral oil, medium chain triglycerides (MCTs), coconut oil; hydrogenated oils, including hydrogenated cottonseed oil, hydrogenated palm oil, hydrogenated castor oil, or hydrogenated soybean oil; derivatives of polyoxyethylene and hydrogenated castor oil, including polyoxyl-40 and hydrogenated castor oil, polyoxyl-60 and hydrogenated castor oil or polyoxyl-100 and hydrogenated castor oil.

In addition, the invention provides methods for modulating kinase activity by contacting a kinase with an amount of a compound of the invention sufficient to modulate kinase activity. Modulation may be inhibition or activation of kinase activity. In some embodiments, the invention provides methods for inhibiting kinase activity by contacting a kinase with an amount of a compound of the invention sufficient to inhibit kinase activity. In some embodiments, the invention provides methods of inhibiting kinase activity in a solution by contacting said solution with an amount of a compound of the invention sufficient to inhibit kinase activity in said solution. In some embodiments, the invention provides methods of inhibiting kinase activity in a cell by contacting said cell with an amount of a compound of the invention sufficient to inhibit kinase activity in said cell. In some embodiments, the invention provides methods of inhibiting kinase activity in a tissue by contacting said tissue with an amount of a compound of the invention sufficient to inhibit kinase activity in said tissue. In some embodiments, the invention provides methods of inhibiting kinase activity in an organism by contacting said organism with an amount of a compound of the invention sufficient to inhibit kinase activity in said organism. In some embodiments, the invention provides methods of inhibiting kinase activity in an animal by contacting said animal with an amount of a compound of the invention sufficient to inhibit kinase activity in said animal. In some embodiments, the invention provides methods of inhibiting kinase activity in a mammal by contacting said mammal with an amount of a compound of the invention sufficient to inhibit kinase activity in said mammal. In some embodiments, the invention provides methods of inhibiting kinase activity in humans by bringing said person into contact with an amount of a compound of the invention sufficient to inhibit kinase activity in said person. In some embodiments, the% kinase activity after contacting the kinase with a compound of the invention is less than 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or 99% of the kinase activity, measured in the absence of this step bringing in contact.

In some embodiments, the kinase is a lipid kinase or protein kinase. In some embodiments, the kinase is selected from the group consisting of PI3 kinase, including various isoforms, such as PI3 kinase α, PI3 kinase β, PI3 kinase γ, PI3 kinase δ; DNA-PK; mTor; Ab, VEGFR, B4 Efrin Receptor (EphB4); TEK receptor tyrosine kinase (TIE2); Fms-like tyrosine kinase 3 (FLT-3); platelet growth factor receptor (PDGFR); RET; ATM ATR; hSmg-1; Hck; Src; epidermal growth factor receptor (EGFR); KIT (stem cell growth factor receptor); insulin receptor (IR) and IGFR (insulin-like growth factor receptor).

In addition, the invention provides methods for modulating PI3 kinase activity by contacting a PI3 kinase with an amount of a compound of the invention sufficient to modulate PI3 kinase activity. Modulation may be inhibition or activation of PI3 kinase activity. In some embodiments, the invention provides methods of inhibiting PI3 kinase activity by contacting a PI3 kinase with an amount of a compound of the invention sufficient to inhibit PI3 kinase activity. In some embodiments, the invention provides methods of inhibiting PI3 kinase activity. Such inhibition can occur in solution, in a cell expressing one or more PI3 kinases, in tissue containing a cell expressing one or more PI3 kinases, or in an organism expressing one or more PI3 kinases. In some embodiments, the invention provides methods of inhibiting PI3 kinase activity in an animal (including mammals, for example humans) by contacting said animal with an amount of a compound of the invention sufficient to inhibit PI3 kinase activity in said animal.

Combination treatment

The present invention also provides combination therapy methods in which an agent known to modulate other pathways or other components of the same pathway or even overlapping sets of target enzymes is used in combination with a compound of the present invention or a pharmaceutically acceptable salt thereof, complex ether, prodrug, solvate, hydrate or derivative. In one aspect, such therapy includes, but is not limited to, combining the subject compound with chemotherapeutic agents, therapeutic antibodies, and radiation therapy to provide a synergistic or additive therapeutic effect.

In one aspect, the compounds or pharmaceutical compositions of the present invention may exhibit synergistic or additive efficacy when administered in combination with agents that inhibit the production or activity of IgE. Such a combination may reduce the undesirable effect of a high level of IgE associated with the use of one or more PI3Kδ inhibitors, if such an effect occurs. This can be especially useful in the treatment of autoimmune and inflammatory disorders (AND), such as rheumatoid arthritis. In addition, the administration of the PI3Kδ or PI3Kδ / γ inhibitors of the present invention in combination with mTOR inhibitors can also lead to a synergistic effect through enhanced inhibition of the PI3K pathway.

In a separate but related aspect of the present invention, there is provided a combination treatment of a disease associated with PI3K6, comprising administering a PI3K6 inhibitor and an agent that inhibits the production or activity of IgE. Other typical PI3Kδ inhibitors are applicable to this combination and are described, for example, in US Pat. No. 6,800,620. Such combination treatment is particularly useful for, but is not limited to, treatment of autoimmune and inflammatory diseases (AND), including rheumatoid arthritis.

Agents that inhibit the production of IgE are known in the art and include, but are not limited to, one or more of TEI-9874, 2- (4- (6-cyclohexyloxy-2-naphthyloxy) phenylacetamide) benzoic acid, rapamycin, rapamycin analogs (i.e., rapalogs), TORC1 inhibitors, TORC2 inhibitors, and any other compounds that inhibit mTORC1 and mTORC2. Agents that inhibit IgE activity include, for example, anti-IgE antibodies, such as, for example, omalizumab and TNX-901.

For the treatment of autoimmune diseases, the compounds or pharmaceutical compositions of the invention can be used in combination with commonly prescribed drugs, including but not limited to Enbrel®, Remicade®, Humira®, Avonex® and Rebif®. For the treatment of respiratory diseases, the compounds or pharmaceutical compositions of the invention can be administered in combination with commonly prescribed drugs, including but not limited to Xolair®, Advair®, Singulair® and Spiriva®.

The compounds of the invention may be formulated or administered together with other agents that act to alleviate the symptoms of inflammatory conditions, such as encephalomyelitis, asthma, and other diseases set forth in this application. These agents include non-steroidal anti-inflammatory drugs (NSAIDs), for example, acetylsalicylic acid; ibuprofen; naproxen; indomethacin; nabumeton; tolmetin, etc. Corticosteroids are used to reduce inflammation and suppress the activity of the immune system. The most commonly prescribed drug of this type is prednisone. Chloroquin (aralen) or hydroxychloroquin (plaquenil) can also be very useful for some individuals diagnosed with lupus. They are most often prescribed for skin and joint symptoms of lupus. Azathioprine (imuran) and cyclophosphamide (cytoxan) inhibit inflammation and tend to suppress the immune system. Other agents, such as methotrexate and cyclosporine, are used to regulate lupus symptoms. Anticoagulants are used to prevent rapid blood clotting. The range of anticoagulants varies from very low dose aspirin, which prevents platelet adhesion, to heparin / coumadin.

In one other aspect, the invention also relates to a pharmaceutical composition for inhibiting abnormal cell growth in a mammal, which comprises an amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof in combination with an amount of an anti-cancer agent ( for example, a chemotherapeutic agent). Many chemotherapeutic agents are currently known in the art and can be used in combination with the compounds of the invention.

In some embodiments, the chemotherapeutic agent is selected from the group consisting of mitosis inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antihormones, angiogenesis inhibitors and antiandrogens. Non-limiting examples are chemotherapeutic agents, cytotoxic agents, and small non-peptide molecules such as glivec (imatinib mesylate), velcad (bortezomib), casodex (bicalutamide), press (gefitinib), and adriamycin, as well as many chemotherapeutic agents. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (CYTOXAN ™); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines, such as benzodopa, carboxwon, meturedopa and uredopa; ethyleneimines and methylamelamines, including altretamine, triethylene melamine, triethylene phosphoramide, triethylene thiophosphoramide and trimethylolomelamine; nitrogen mustard analogs such as chlorambucil, chlorofazine, holophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembikhine, phenesterol, prednimustine, trophosphamide, uracil mustard; nitrosoureas such as carmustine, chlorzotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics, such as aclacinomisins, actinomycin, autramycin, azaserin, bleomycin, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, Casodex ™, chromomycin, dactinomycin, daunorubicin, detorubicin-dociborin-diboricin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobisin-diobidein , esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin, quelamycin, rhodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimerexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, karmofur, cytarabine, dideoxyuridine, doxyfluuridine, enocytabine, phloxuridine; androgens such as calusterone, dromostanolone propionate, epithiostanol, mepitiostan, testolactone; antiadrenal agents such as aminoglutethimide, mitotan, trilostane; a folic acid replenisher such as folinic acid; Aceglaton; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisanthrene; edatraxate; defofamine; demecolcin; diaziquon; elfomitin; elliptinium acetate; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitocasone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pyrarubicin; podophyllic acid; 2-ethyl hydrazide; procarbazine; PSK.R ™; razoxane; sisofiran; spiro germanium; tenuazonic acid; triaziquon; 2,2 ', 2' '- trichlorotriethylamine; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacitosin; arabinoside (“Ara-C ''); cyclophosphamide; thiotepa; taxanes, e.g. paclitaxel (TAXOL ™, Bristol-Myers Squibb Oncology, Princeton, NJ) and docetaxel (TAXOTERE ™, Rhone-Poulenc Rorer, Antony, France); retinoic acid; esperamycins; capecitabine; and pharmaceutically acceptable salts, acids, or derivatives of any of the above.Anti-hormonal agents that act by regulating or inhibiting the hormonal activity of tumors, such as antiestrogens, are also included as substances suitable for improving the condition of cells during chemotherapy. including, for example, tamoxifen (Nolvadex ), raloxifene, aromatase inhibitors of 4 (5) -imidazoles, 4-hydroxy tamoxifen, trioxifene, keoxifen, LY 117018, onapriston and toremifene (fareston); and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide gemielecilebide goselimerbimer ilecelide; 6-thioguanine; mercaptopurine; methotrexate; platinum analogues such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; Navelbin; novantron; teniposide; daunomycin; aminopterin; xeloda; ibandronate; Captothecin-11 (SRT-11); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO). Where desired, the compounds or pharmaceutical composition of the present invention can be used in combination with commonly prescribed anti-cancer drugs such as Herceptin®, Avastin®, Erbitux®, Rituxan®, Taxol®, Arimidex®, Taxotere® and Velcade® .

The invention also relates to a method of using the compounds or pharmaceutical composition in combination with radiation therapy to inhibit abnormal cell growth, or to a method of treating a hyperproliferative disorder in a mammal. Techniques for applying radiation therapy are known in the art, and these techniques can be used in the combination therapy set forth in this application. The administration of a compound of the invention in such combination therapy can be determined as set forth in this application.

Radiation therapy can be carried out using one of several methods or a combination of methods, including, without limitation, external remote radiation therapy, internal radiation therapy, implant radiation, stereotactic radiosurgery, systemic radiation therapy, radiotherapy, and long or short-term interstitial brachytherapy. The term "brachytherapy" as used in this application refers to radiation therapy carried out by means of a spatially limited radioactive substance introduced into or near the tumor site or to another place of a proliferative tissue disease. The term is intended to include, without limitation, exposure to radioactive isotopes (e.g., At-211, I-131, 1-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32 and radioactive isotopes Lu). Suitable radiation sources for use as the cell-improving substance of the present invention include both solids and liquids. By way of non-limiting example, the radiation source may be a radionuclide, such as 1-125, 1-131, Yb-169, lr-192 as a solid source, 1-125 as a solid source, or other radionuclides that emit protons, beta particles, gamma radiation or other types of radiation used in therapy. In addition, the radioactive substance may be a liquid prepared from any solution of radionuclide (s), for example, a solution of 1-125 or 1-131, or a radioactive liquid can be prepared using a suspension of a suitable liquid containing small particles of solid radionuclides, such as Au- 198, Y-90. In addition, the radionuclide (s) may be included in the gel or be part of the radioactive microspheres.

Without limiting itself to any theory, it can be said that the compounds of the present invention can make abnormal cells more sensitive to radiation treatment in order to destroy and / or inhibit the growth of such cells. Accordingly, this invention also relates to a method for sensitizing abnormal cells in a mammal to radiation treatment, comprising administering to the mammal an amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof that is effective for sensitizing abnormal cells to treatment irradiation. The amount of compound, salt or solvate in this method can be determined in accordance with methods for determining effective amounts of such compounds set forth in this application.

The compounds or pharmaceutical compositions of the present invention can be used in combination with an amount of one or more substances selected from antiangiogenetic agents, signal transduction inhibitors, and antiproliferative agents.

Antiangiogenetic agents such as MMP-2 inhibitors (matrix metalloproteinases 2), MMP-9 inhibitors (matrix metalloproteinases 9) and COX-11 inhibitors (cyclooxygenase 11) can be used together with the compound of the present invention and the pharmaceutical compositions described herein application. Examples of useful COX-P inhibitors include CELEBREX ™ (alecoxib), valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in WO 96/33172 (published October 24, 1996), WO 96/27583 (published March 7, 1996), European patent application No. 97304971.1 (filed July 8

1997), European Patent Application No. 99308617.2 (filed October 29, 1999), WO 98/07697 (published February 26, 1998), WO 98/03516 (published January 29, 1998), WO 98/34918 (published August 13, 1998), WO 98/34915 (published August 13, 1998), WO 98/33768 (published August 6, 1998), WO 98/30566 (published July 16, 1998), European publication patent 606046 (published July 13, 1994), publication of European patent 931788 (published July 28, 1999), WO 90/05719 (published May 31, 1990), WO 99/52910 (published October 21, 1999), WO 99/52889 (published October 21, 1999), WO 99/2966 7 (published June 17, 1999), PCT International Application No.PCT / IB98 / 01113 (filed July 21, 1998), European Patent Application No. 99302232.1 (filed March 25, 1999), UK Patent Application No. 9912961.1 ( filed June 3, 1999), provisional application US No. 60/148464 (filed August 12, 1999), US patent 5863949 (published January 26, 1999), US patent 5861510 (published January 19, 1999) and European publication Patent 780386 (published June 25, 1997), all of which are incorporated herein by reference in their entireties. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity with respect to inhibition of MMP-1. More preferred are those that selectively inhibit MMP-2 and / or AMP-9 compared to other matrix metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12 and MMP-13). Some specific examples of MMP inhibitors useful in the present invention are AG-3340, RO 32-3555 and RS 13-0830.

The invention also relates to a method of treatment and to a pharmaceutical composition for treating a cardiovascular disease in a mammal, comprising an amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof or an isotope-labeled derivative thereof, and an amount of one or more therapeutic agents used to treat cardiovascular disease.

Examples for use in relation to cardiovascular diseases are antithrombotic agents, for example, prostacyclin and salicylates, thrombolytic agents, for example, streptokinase, urokinase, tissue plasminogen activator (TPA), and an anisoylated activator complex of plasminogen and streptokinase (APSAC), antithrombotic, acetylsalicylic acid (ASA) and clopidrogel, vasodilators, e.g. nitrates, calcium channel blocking drugs, antiproliferative agents, e.g. olchicine and alkylating agents, intercalating agents, growth modulating factors such as interleukins, transforming growth factor beta and platelet related growth factor compounds, anti-growth factor monoclonal antibodies, anti-inflammatory agents, both steroidal and non-steroidal agents, and other agents that can modulate vascular tone, function, arteriosclerosis, and the healing response to damage to blood vessels or organs after exposure. According to the invention, antibiotics may also be included in the composition of the combinations or substances for coating. Moreover, the coating substance can be used to effect therapeutic delivery to the focus in the vessel wall. By incorporating the active agent into the swellable polymer, the active agent will be released as the polymer swells.

The compounds described in this application can be manufactured and introduced together with substances representing liquid or solid tissue barriers, also known as lubricants. Examples of tissue barriers include, but are not limited to, polysaccharides, politicans, seprafilm, interseed, and hyaluronic acid.

Medicines that can be administered together with the compounds set forth in this application include any suitable medications that are useful by inhalation, for example, analgesics, for example codeine, dihydromorphine, ergotamine, fentanyl or morphine; anginal preparations, for example, diltiazem; antiallergic agents, for example, cromoglycate, ketotifen or nedocromil; anti-infective agents, for example, cephalosporins, penicillins, streptomycin, sulfonamides, tetracyclines or pentamidine; antihistamines, for example, metapyrene; anti-inflammatory drugs, for example, beclomethasone, flunisolide, budesonide, tipredan, triamcinolone acetonide or fluticasone; antitussive agents, for example, noscapine; bronchodilators, for example, ephedrine, adrenaline, fenoterol, formoterol, isoprenaline, metaproterenol, phenylephrine, phenylpropanolamine, pirbuterol, reproterol, rimeterol, salbutamol, salmeterol, terbutaline, isoetarin, orulocyrenol-4-aminoterolene dichloro-α - [[[6- [2- (2-pyridinyl) ethoxy] hexyl] amino] methyl] benzenemethanol; diuretics, for example amiloride; anticholinergics, for example, ipratropium, atropine or oxytropium; hormones, for example, cortisone, hydrocortisone or prednisone; xanthines, for example, aminophylline, choline theophyllinate, lysine theophyllinate or theophylline; and therapeutic proteins and peptides, for example, insulin or glucagon. It will be apparent to those skilled in the art that, where appropriate, drugs can be used in the form of salts (for example, as alkali metal or amine salts or as acid addition salts) or as esters (e.g. lower alkyl esters) or as solvates (e.g. hydrates) to optimize the activity and / or stability of the drug.

Other typical therapeutic agents useful for combination therapy include, but are not limited to, the agents described above, radiation therapy, hormone antagonists, hormones and their releasing factors, thyroid and antithyroid drugs, estrogens and progestins, androgens, adrenocorticotropic hormone;

adrenocorticosteroids and their synthetic analogues; inhibitors of the synthesis and action of hormones of the adrenal cortex, insulin, oral hypoglycemic agents and pharmacology of the endocrine pancreas; agents for calcification and bone remodeling: calcium, phosphate, parathyroid hormone, vitamin D, calcitonin, vitamins such as water-soluble vitamins, vitamin B complex, ascorbic acid, fat-soluble vitamins, vitamins A, K and E, growth factors, cytokines chemokines, agonists and antagonists of muscarinic receptors; anticholinesterase agents; agents acting on the neuromuscular synapse and / or autonomic ganglion; catecholamines, sympathomimetic drugs and adrenergic receptor agonists or antagonists; and 5-hydroxytryptamine receptor agonists and antagonists (5-HT, serotonin).

Therapeutic agents may also include pain and inflammation agents such as histamine and histamine antagonists, bradykinin and bradykinin antagonists, 5-hydroxytryptamine (serotonin), lipid substances created by biotransformation of products of selective hydrolysis of membrane phospholipids, eicosanoids, prostaglandins, prostaglandins, prostaglandins aspirin, non-steroidal anti-inflammatory agents, analgesics, antipyretic drugs, agents inhibiting the synthesis of prostaglandins and thromboxanes, selective inhibitors of cibel cyclooxygenase, selective inhibitors of inducible cyclooxygenase-2, autacoids, paracrine hormones, somatostatin, gastrin, cytokines, mediating interactions involved in the humoral and cellular immune responses, lipid autacoids, eicosanoids, P-adrenergic agonists, methyl antioxidants, and sodium channels, opioid receptor agonists, calcium channel blockers, membrane stabilizers and leukotriene inhibitors.

Additional therapeutic agents contemplated herein include diuretics, vasopressin, kidney water retention agents, rennin, angiotensin, agents useful in the treatment of myocardial ischemia, antihypertensive agents, angiotensin converting enzyme inhibitors, β-adrenergic receptor antagonists, for the treatment of hypercholesterolemia and agents for the treatment of dyslipidemia.

Other contemplated therapeutic agents include drugs used to regulate the acidity of the gastrointestinal tract, drugs for treating peptic ulcers, drugs for treating gastroesophageal reflux disease, prokinetic agents, antiemetics, drugs used for irritable bowel syndrome, drugs used for diarrhea, remedies for constipation, remedies for inflammatory bowel disease, remedies for bile duct disease pathways, drugs used for pancreatic disease. Therapeutic agents used to treat protozoal infections, drugs used to treat malaria, amebiasis, giardiasis, trichomoniasis, trypanosomiasis and / or leishmaniasis, and / or drugs used in chemotherapy for helminthiasis. Other therapeutic agents include antimicrobial agents, sulfonamides, trimethoprim-sulfamethoxazole quinolones and agents for urinary tract infections, penicillins, cephalosporins and others, β-lactam antibiotics, an agent containing aminoglycoside, protein synthesis inhibitors, drugs used in chemotherapy for tuberculosis caused by Mycobacterium avium, and leprosy, antifungal agents, antiviral agents, including non-retroviral agents and antiretroviral agents.

Examples of therapeutic antibodies that can be combined with the compound of the invention include, but are not limited to, antibodies against receptor tyrosine kinases (cetuximab, panitumumab, trastuzumab), antibodies against CD20 (differentiation cluster 20) (rituximab, tositumomab) and other antibodies such as alemtuzumab, bevacizumab and gemtuzumab.

In addition, the methods described in this application provide therapeutic agents used for immunomodulation, such as immunomodulators, immunosuppressive agents, tolerogenes and immunostimulants. In addition, therapeutic agents affecting the blood and blood forming organs, hematopoietic agents, growth factors, minerals and vitamins, anticoagulant, thrombolytic and antiplatelet drugs.

Additional therapeutic agents that can be combined with the compound of the invention can be found in Goodman and Gilman's book "The Pharmacological Basis of Therapeutics", tenth edition, edited by Hardman, Limbird and Gilman, or in the Physician's Desk Reference ); both are included in this description in its entirety by reference.

The compounds of this application can be used in combination with the agents described in this application or other suitable agents, depending on the condition being treated. In other words, in some embodiments, the compounds of the invention will be administered together with other agents as described above. When used in combination therapy, the compounds described in this application can be administered together with the second agent simultaneously or separately. Such administration in combination may include simultaneous administration of two agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, the compound set forth in this application and any of the agents described above can be formulated together in the same dosage form and administered simultaneously. Alternatively, the compound of the present invention and any of the agents described above can be administered simultaneously, both agents being present in separate compositions. In another alternative, any of the agents described above can be administered immediately after the compound of the present invention, or vice versa. According to the separate administration protocol, the compound of the present invention and any of the agents described above can be administered with an interval of several minutes, or with an interval of several hours, or with an interval of several days.

The examples and preparatory examples presented below further illustrate the compounds of the present invention and methods for producing such compounds and are given as an example. It should be understood that the scope of the present invention is in no way limited by the scope of the following examples and preparatory examples. In the following examples, molecules with a single chiral center, unless otherwise indicated, exist as a racemic mixture. Such molecules with two or more chiral centers, unless otherwise indicated, exist as a racemic mixture of diastereomers. Single enantiomers / diastereomers can be obtained by methods known to specialists in this field of technology.

EXAMPLES

Example 1. Synthesis of 3 - ((4-amino-3- (3-hydroxyphenyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (Compound 1613) (method A).

Scheme 12. The synthesis of 3 - ((4-amino-3- (3-hydroxyphenyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinoline- 1 (2H) -one (Compound 1613).

A solution of 2-amino-6-methylbenzoic acid (1601) (106.5 g; 705 mmol) in H ₂ O (200 ml) was cooled to 0-5 ° C, concentrated HCl (250 ml) was slowly added. The solution was stirred for 15 min at 0-5 ° C. A solution of sodium nitrite (58.4 g; 6.85 mol) in H ₂ O (120 ml) was added dropwise at 0-5 ° C and the resulting mixture was stirred for 30 minutes. Then, the above solution was added to a solution of KI (351 g; 2.11 mol) in H ₂ O (200 ml) and the resulting mixture was stirred at RT for 16 hours. The solution was poured into ice water (2000 ml) and extracted with ethyl acetate (3 × 1000 ml). The combined organic layer was washed with aqueous NaOH (15%, 3 × 200 ml). The aqueous layer was acidified to pH 1 and extracted with ethyl acetate (3 × 1000 ml). The combined organic layers were dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo, which afforded the desired product 2-iodo-6-methylbenzoic acid (1602) (145 g; yield 79%) as a yellow solid.

To a stirred mixture of 2-iodo-6-methylbenzoic acid (1602) (105 g; 400 mmol), Pd (OAc) 2 (27 g; 120 mmol) and PPh ₃ (63 g; 240 mol) in THF (1000 ml) at CT, tributyl (vinyl) tin (152 g; 480 mmol) was added. The resulting mixture was heated to reflux overnight. The mixture was allowed to cool to RT, filtered through silica gel (10 g) and then concentrated in vacuo. The residue was poured into ice water (1000 ml) and extracted with ethyl acetate (3 × 1000 ml). The combined organic layer was washed with aqueous NaOH (15%, 5 × 200 ml). The combined aqueous layer was acidified to pH 1, extracted with ethyl acetate (3 × 1000 ml). The combined organic layer was dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo, which afforded the desired product 2-methyl-6-vinylbenzoic acid (1603) (61 g; 95% yield) as a yellow solid.

A mixture of 2-methyl-6-vinylbenzoic acid (1603) (56 g; 350 mmol) and thionyl chloride (208 g; 1750 mmol) in toluene (400 ml) was stirred at reflux for 2 hours. The mixture was concentrated in vacuo, which allowed to obtain the desired product 2-methyl-6-vinylbenzoyl chloride (1604) (63 g; 95% yield) as a yellow oil. The resulting product was used directly in the next step without purification.

A mixture of o-toluidine (45 g; 420 mmol) and triethylamine (71 g; 70 mmol) in CH ₂ Cl ₂ (300 ml) was stirred for 10 minutes at RT. To this mixture was added 2-methyl-6-vinylbenzoyl chloride (1604) (63 g; 35 mmol) and the resulting mixture was stirred at RT for 30 minutes. The solution was poured into water (300 ml), extracted with CH ₂ Cl ₂ (3 × 200 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo, which allowed to obtain a crude product. The crude product was suspended in IPE (isopropyl ether) (300 ml), stirred at reflux for 30 minutes and then cooled to 0-5 ° C. The precipitate was collected by filtration and then dried in vacuo, which afforded the desired product 2-methyl-N-o-tolyl-6-vinylbenzamide (1605) (81 g; yield 80%) as a yellow solid.

To a solution of 2-methyl-N-o-tolyl-6-vinylbenzamide (1605) (80 g; 320 mmol) in DMF (250 ml) at RT, NaH (60% in mineral oil; 25.6 g; was slowly added). 640 mmol) and the resulting mixture was stirred at RT for 30 minutes. Ethyl chloroacetate (78 g; 640 mmol) was added to this mixture and the resulting mixture was stirred at RT for 2 hours. The solution was poured into water (500 ml), extracted with ethyl acetate (3 × 200 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo. The crude product was suspended in MeOH (160 ml), stirred at reflux for 10 minutes and then cooled to 0-5 ° C. The precipitate was collected by filtration and then dried in vacuo, which afforded the desired product ethyl 2- (2-methyl-N-o-tolyl-6-vinylbenzamido) acetate (1606) (67 g; yield 62%) as a white solid .

To a stirred mixture of ethyl 2- (2-methyl-N-o-tolyl-6-vinylbenzamido) acetate (1606) (67 g; 200 mmol) in 1,4-dioxane (300 ml) and H ₂ O (100 ml ) osmium tetroxide (20 mg) was added at RT and stirred at RT for 30 min. Sodium periodate (86 g; 400 mmol) was added to this mixture and the resulting mixture was stirred at RT for 16 hours. The reaction mixture was filtered through silica gel (10 g), the filtrate was extracted with ethyl acetate (3 × 200 ml). The combined organic layers were washed with brine (100 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo and the residue was then dried in vacuo, which afforded the desired product ethyl 2- (2-formyl-6-methyl-N-o-tolylbenzamido) acetate (1607) (38 g); yield 57%) as yellow solid matter.

To a stirred solution of ethyl 2- (2-formyl-6-methyl-N-o-tolylbenzamido) acetate (1607) (38 g; 112 mmol) in EtOH (200 ml) and ethyl acetate (100 ml), cesium carbonate was added at RT (22 g; 112 mmol). The resulting mixture was degassed, filled again with argon three times and then stirred at 50 ° C for 5 hours. The mixture was allowed to cool to RT, filtered through silica gel (10 g), and the filtrate was concentrated in vacuo. The residue was poured into H ₂ O (200 ml), extracted with ethyl acetate (3 × 200 ml). The combined organic layer was washed with brine (50 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo. The crude product was suspended in IPE (120 ml), heated to reflux for 10 minutes, and then cooled to 0-5 ° C. The precipitate was collected by filtration and then dried in vacuum, which allowed to obtain the desired product ethyl-8-methyl-1-oxo-2-o-tolyl-1,2-dihydroisoquinoline-3-carboxylate (1608) (28 g; yield 77%) in the form of a white solid.

To a stirred solution of lithium aluminum hydride (8.28 g; 218 mol) in anhydrous THF (500 ml) at -78 ° C under nitrogen atmosphere, ethyl 8-methyl-1-oxo-2-o-tolyl was slowly added over 10 minutes -1,2-dihydroisoquinoline-3-carboxylate (1608) (28 g; 87 mmol). The resulting mixture was allowed to warm to −30 ° C., stirred for 30 minutes, and TLC showed that the reaction was complete. The mixture was then cooled to -78 ° C and water (50 ml) was slowly added. The mixture was allowed to warm to RT, filtered through silica gel (10 g) and the filtrate was concentrated in vacuo. The crude product was poured into H ₂ O (200 ml) and extracted with ethyl acetate (3 × 200 ml). The combined organic layer was washed with brine (50 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo. The crude product was suspended in ethyl acetate (30 ml) and stirred for 10 minutes. The solid was collected by filtration and then dried in vacuo, which afforded the desired product 3- (hydroxymethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (1609) (22 g; yield 92%) as white solid.

PBr ₃ (25.6 g; 95 mmol) was slowly added to a stirred solution of DMF (11.5 g; 158 mol) in acetonitrile (200 ml) at 0 ° C and the resulting mixture was stirred at 0 ° C for 30 minutes. 3- (hydroxymethyl) -8-methyl-2-o-tolylisoquinolin-1- (2H) -one (1609) (22 g; 78.8 mmol) was slowly added. Then the reaction mixture was allowed to warm to RT and stirred for 30 minutes. Saturated aqueous NaHCO ₃ solution (50 ml) was slowly added and extracted with ethyl acetate (3 × 200 ml). The combined organic layer was washed with brine, dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo. The crude product was suspended in IPE (50 ml) and then stirred for 10 minutes. The precipitate was collected by filtration and then dried in vacuo, which afforded the desired product 3- (bromomethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (1610) (21 g; yield 80%) as white solid matter.

3-Iodo-1H-pyrazolo [3,4-d] pyrimidin-4-amine (108) (10.8 g; 41.4 mmol) and potassium tert-butylate (4.4 g; 40 mmol) were dissolved in anhydrous DMF (150 ml) and stirred at RT for 30 minutes. 3- (Bromomethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (1610) (13.7 g; 40 mmol) was added. The resulting mixture was stirred at RT for 30 min, poured into ice water (300 ml) and then extracted with ethyl acetate (3 × 200 ml). The combined organic layer was washed with brine (50 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated to about 100 ml in vacuo, the precipitate was collected by filtration, which allowed to obtain the first batch of the desired product 3 - ((4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (1611) (12 g; 60% yield) as a white solid. The filtrate was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (2-20% MeOH / DCM), which allowed to obtain a second batch of the desired product 3 - ((4-amino-3-iodo-1H-pyrazolo [3,4- d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (1611) (6 g; yield 30%) as a white solid.

3 - ((4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (1611) (13 g; 24.9 mmol) and 3- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenol (1612) (6.6 g; 30 mmol) were dissolved in mixtures of DMF-EtOH-H ₂ O (120 ml; 40 ml; 40 ml). Pd (OAc) 2 (1.684 g; 7.5 mmol), PPh ₃ (3.935 g; 15 mmol) and Na ₂ CO ₃ (13.25 g; 125 mmol) were successively added. The resulting mixture was degassed, filled again with argon three times, and then stirred at 100 ° C for 1 h. The mixture was allowed to cool to RT, filtered through silica gel (10 g) and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (2-20% MeOH / DCM), which allowed to obtain the product (1613) (9 g; yield 76%) as a light yellow solid. Then, the above product was suspended in EtOH (100 ml) and heated to reflux for 30 minutes. The mixture was allowed to cool to RT and the solid was collected by filtration. The solid was then suspended in EA (ethyl acetate) (100 ml) and stirred overnight. The precipitate was collected by filtration, then dried in vacuo, which afforded the desired product 3 - ((4-amino-3- (3-hydroxyphenyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8- methyl-2-o-tolylisoquinolin-1 (2H) -one (1613) (8.4 g; yield 69%) as a white solid.

Example 2. Synthesis of 3 - ((4-amino-3- (3-hydroxyphenyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (Compound 1613) (method B).

Scheme 13. The synthesis of 3 - ((4-amino-3- (3-hydroxyphenyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinoline- 1 (2H) -one (Compound 1613) by Method B.

3- (3-Methoxyphenyl) -1H-pyrazolo [3,4-d] pyrimidin-4-amine (1701) (964 mg; 4 mmol) and potassium tert-butylate (0.44 g; 4 mmol) were dissolved in anhydrous DMF (150 ml) and stirred at RT for 30 minutes. 3- (Bromomethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (1610) (1.37 g; 4.0 mmol) was added. The resulting mixture was stirred at RT for 30 min, poured into ice water (30 ml) and then extracted with ethyl acetate (3 × 50 ml). The combined organic layer was washed with brine (25 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (2-20% MeOH / DCM), which allowed us to obtain the desired product 3 - ((4-amino-3- (3-methoxyphenyl) -1H-pyrazolo [3, 4-d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (1702) (1.4 g; yield 70%) as a white solid.

To a solution of 3 - ((4-amino-3- (3-methoxyphenyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H ) -one (1702) (100 mg; 0.2 mmol) in CH ₂ Cl ₂ (20 ml) at -78 ° C under nitrogen atmosphere was added BBr ₃ (1 ml) and the resulting mixture was stirred at -78 ° C for 3 h. The mixture was allowed to warm to RT, poured into ice-water (200 ml) and extracted with ethyl acetate (3 × 50 ml). The combined organic layer was washed with brine (20 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (10-50% MeOH / CH ₂ Cl ₂ ), which allowed to obtain the desired product 3 - ((4-amino-3- (3-hydroxyphenyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (1613) (87 mg; yield 91%) as a white solid.

Example 3. Synthesis of (R) -3 - ((4-amino-3- (3-hydroxybut-1-ynyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8-methyl -2-o-tolylisoquinolin-1 (2H) -one (Compound 1802).

Scheme 14. The synthesis of (R) -3 - ((4-amino-3- (3-hydroxybut-1-ynyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8- methyl-2-o-tolylisoquinolin-1 (2H) -one (Compound 1802).

3 - ((4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (1611) (522 mg; 1 mmol) and (R) -but-3-yn-2-ol (84 mg; 1.2 mmol) were dissolved in anhydrous THF (40 ml). The mixture was degassed and filled again with nitrogen three times. Pd (PPh ₃ ) ₂ Cl ₂ (12 mg; 0.1 mmol), CuI (47 mg; 0.25 mmol) and (iso-Pr) ₂ NH (505 mg; 5 mmol) were successively added. The resulting mixture was degassed, filled again with argon three times and then stirred at reflux for 4 hours. The mixture was allowed to cool to RT, filtered through silica gel (10 g) and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (2-20% MeOH / DCM), which allowed to obtain the product 3 (R) -3 - ((4-amino-3- (3-hydroxybut-1-ynyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (1802) (324 mg; 70% yield) as a light yellow solid .

Example 4. Synthesis of 3 - ((6-amino-9H-purin-9-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (Compound 1902).

Scheme 15. The synthesis of 3 - ((6-amino-9H-purin-9-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one is described (Compound 1902).

9H-Purin-6-amine (1901) (540 mg; 4.0 mmol) was dissolved in anhydrous DMF (20 ml). NaH (60% in mineral oil; 160 mg; 4.0 mmol) was added and the resulting mixture was stirred at RT for 30 minutes. 3- (Bromomethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (1610) (1.37 g; 4.0 mmol) was added. The reaction mixture was stirred at RT for 30 min, poured into ice-water (30 ml) and then extracted with ethyl acetate (3 × 50 ml). The combined organic layer was washed with brine (25 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (2-20% MeOH / DCM), which afforded the desired product 3 - ((6-amino-9H-purin-9-yl) methyl) -8-methyl -2-o-tolylisoquinolin-1 (2H) -one (1902) (1.1 g; 70% yield) as a white solid.

Example 5. Synthesis of 3 - ((4-amino-3- (3-hydroxyphenyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -2-isopropyl-8-methylisoquinolin-1 (2H ) -one (Compound 2013).

Scheme 16. The synthesis of 3 - ((4-amino-3- (3-hydroxyphenyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -2-isopropyl-8-methylisoquinolin-1 ( 2H) -one (Compound 2010).

To a stirred mixture of 2-iodo-6-methylbenzoic acid (1602) (105 g; 400 mmol), Pd (OAc) 2 (27 g; 120 mmol) and PPh ₃ (63 g; 240 mol) in THF (1000 ml) at CT, tributyl (vinyl) tin (152 g; 480 mmol) was added. The resulting mixture was heated to reflux overnight. The mixture was allowed to cool to RT, filtered through silica gel (10 g) and then concentrated in vacuo. The residue was poured into ice water (1000 ml) and extracted with ethyl acetate (3 × 1000 ml). The combined organic layer was washed with aqueous NaOH (15%; 5 × 200 ml). The combined aqueous layer was acidified to pH 1, extracted with ethyl acetate (3 × 1000 ml). The combined organic layer was dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo, which afforded the desired product 2-methyl-6-vinylbenzoic acid (1603) (61 g; 95% yield) as a yellow solid.

A mixture of 2-methyl-6-vinylbenzoic acid (1603) (56 g; 350 mmol) and thionyl chloride (208 g; 1750 mmol) in toluene (400 ml) was stirred at reflux for 2 hours. The mixture was concentrated in vacuo, which allowed to obtain the desired product 2-methyl-6-vinylbenzoyl chloride (1604) (63 g; 95% yield) as a yellow oil. The resulting product was used directly in the next step without purification.

Propan-2-amine (2001) (59 g; 1.0 mol) and ethyl chloroacetate (122 g; 1.0 mol) were dissolved in toluene (200 ml) and the mixture was stirred at reflux for 2 hours. The reaction mixture was allowed to cool to CT, poured into ice-water (500 ml) and extracted with ethyl acetate (3 × 250 ml). The combined organic layer was washed with brine (50 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (10-50% EA / PE), which afforded the product ethyl 2- (isopropylamino) acetate (2002) (70 g; 51% yield) as an oil.

Ethyl 2- (isopropylamino) acetate (2002) (14.5 g; 100 mmol) and triethylamine (200 g; 200 mmol) were dissolved in CH ₂ Cl ₂ (300 ml) and the mixture was stirred for 10 minutes at RT. 2-Methyl-6-vinylbenzoyl chloride (1604) (18 g; 100 mmol) was added and the resulting mixture was stirred at RT for 30 minutes. The reaction mixture was poured into water (300 ml) and extracted with CH ₂ Cl ₂ (3 × 200 ml). The combined organic layer was washed with brine (50 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo, which allowed to obtain a crude product. The crude product was suspended in IPE (isopropyl ether) (300 ml), stirred at reflux for 30 minutes and then cooled to 0-5 ° C. The precipitate was collected by filtration and then dried in vacuo, which afforded the desired product ethyl 2- (N-isopropyl-2-methyl-6-vinylbenzamido) acetate (2003) (14.5 g; yield 50%) as a yellow solid substances.

To a stirred solution of ethyl 2- (N-isopropyl-2-methyl-6-vinylbenzamido) acetate (2003) (14.0 g; 48.0 mmol) in 1,4-dioxane (100 ml) and H ₂ O ( 30 ml) osmium tetroxide (20 mg) was added and the resulting mixture was stirred at RT for 30 minutes. Sodium periodate (22 g; 100 mmol) was added to this mixture and then stirred at RT for 16 hours. The reaction mixture was filtered through silica gel (10 g), the filtrate was extracted with ethyl acetate (3 × 200 ml). The combined organic layer was washed with brine (50 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo and the residue was then dried in vacuo, which afforded the desired product ethyl 2- (2-formyl-N-isopropyl-6-methylbenzamido) acetate (2004) (8.33 g; yield 57%) as yellow solid matter.

To a stirred solution of ethyl 2- (2-formyl-N-isopropyl-6-methylbenzamido) acetate (2004) (8.3 g; 28.0 mmol) in EtOH (100 ml) and ethyl acetate (50 ml) at RT was added cesium carbonate (5.9 g; 30 mmol). The resulting mixture was degassed, filled again with argon three times and then stirred at 50 ° C for 5 hours. The mixture was allowed to cool to RT, filtered through silica gel (10 g), and the filtrate was concentrated in vacuo. The residue was poured into H ₂ O (200 ml), extracted with ethyl acetate (3 × 200 ml). The combined organic layer was washed with brine (50 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo. The crude product was suspended in IPE (120 ml), stirred at reflux for 10 minutes and then cooled to 0-5 ° C. The precipitate was collected by filtration and then dried in vacuo, which afforded the desired product ethyl-2-isopropyl-8-methyl-1-oxo-1,2-dihydroisoquinoline-3-carboxylate (2005) (5.35 g; 70% yield) in the form of a white solid.

To a stirred solution of lithium aluminum hydride (2.88 g; 76 mol) in anhydrous THF (200 ml) at -78 ° C, ethyl 2-isopropyl-8-methyl-1-oxo-1 was added slowly over a period of 10 minutes under a nitrogen atmosphere. , 2-dihydroisoquinoline-3-carboxylate (2005) (5.2 g; 19 mmol). The resulting mixture was allowed to warm to −30 ° C., stirred for 30 minutes, and TLC showed that the reaction was complete. Then the mixture was cooled to −78 ° C. Water (50 ml) was slowly added. The mixture was allowed to warm to RT, filtered through silica gel (10 g) and the filtrate was concentrated in vacuo. The crude product was poured into H ₂ O (200 ml) and extracted with ethyl acetate (3 × 200 ml). The combined organic layer was washed with brine (50 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo. The crude product was suspended in ethyl acetate (30 ml) and stirred for 10 minutes. The solid was collected by filtration, then dried in vacuo, which afforded the desired product 3- (hydroxymethyl) -2-isopropyl-8-methylisoquinolin-1 (2H) -one (2006) (3.51 g; yield 80%) as white solid.

To a solution of 3- (hydroxymethyl) -2-isopropyl-8-methylisoquinolin-1 (2H) -one (2006) (1.61 g; 7.0 mmol) in CH ₂ Cl _{2 was} added PPh ₃ (3.67 g; 14.0 mmol) and the mixture was stirred at RT for 30 minutes. The mixture was cooled to 0 ° C and CBr ₄ (4.64 g; 14.0 mmol) was added portionwise. The resulting mixture was stirred at a temperature of from 0 ° C to CT for 30 minutes and then concentrated in vacuo. The crude product was purified by flash column chromatography on silica gel (30-50% EA / PE), which allowed us to obtain the desired product 3- (bromomethyl) -2-isopropyl-8-methylisoquinolin-1 (2H) -one (2007) (1, 65 g; 80% yield) as a white solid.

A mixture of 3-iodo-1H-pyrazolo [3,4-d] pyrimidin-4-amine (108) (1.3 g; 5 mmol) and potassium tert-butylate (0.55 g; 5 mmol) in anhydrous DMF ( 20 ml) was stirred at RT for 30 min and then 3- (bromomethyl) -2-isopropyl-8-methylisoquinolin-1 (2H) -one (2007) (1.47 g; 5 mmol) was added. The resulting mixture was stirred at RT for 30 min, poured into ice-water (30 ml) and then extracted with ethyl acetate (3 × 50 ml). The combined organic layer was washed with brine (25 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (2-20% MeOH / DCM), which afforded the desired product 3 - ((4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -2-isopropyl-8-methylisoquinolin-1 (2H) -one (2008) (1.66 g; yield 70%) as a white solid.

To a stirred mixture of 3 - ((4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -2-isopropyl-8-methylisoquinolin-1 (2H) -one (2008 ) (95 mg; 0.2 mmol) and 3- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenol (66 mg; 0.3 mmol) in a mixture of DMF- EtOH-H ₂ O (3: 1: 1; 20 ml) was sequentially added Pd (OAc) 2 (16 mg; 0.075 mmol), PPh ₃ (39.3 mg; 0.15 mmol) and Na ₂ CO ₃ (132 mg; 1.25 mmol). The resulting mixture was degassed, filled again with argon three times, and then stirred at 100 ° C for 1 h. The mixture was allowed to cool to RT, filtered through silica gel (10 g) and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (2-20% MeOH / DCM), which afforded the product 3 - ((4-amino-3- (3-hydroxyphenyl) -1H-pyrazolo [3,4-d] pyrimidine- 1-yl) methyl) -2-isopropyl-8-methylisoquinolin-1 (2H) -one (2009) (53 mg; 61% yield) as a pale yellow solid.

Example 6. Synthesis of 8-methyl-3 - ((methyl (9H-purin-6-yl) amino) methyl) -2-o-tolylisoquinolin-1 (2H) -one.

Scheme 17. The synthesis of 8-methyl-3 - ((methyl (9H-purin-6-yl) amino) methyl) -2-o-tolylisoquinolin-1 (2H) -one (Compound 4004) is described.

3- (Bromomethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (342 mg; 1.0 mmol) (1610) was dissolved in methylamine solution (100 ml) and stirred for 2 hours. The mixture poured into a mixture of ice in water (200 ml) and extracted with ethyl acetate (3 × 50 ml). The combined organic layer was washed with brine (20 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo, which afforded the desired product 8-methyl-3 - ((methylamino) methyl) -2-o-tolylisoquinolin-1 (2H) -one (4001) (250 mg; 86% yield) as a yellow solid substances. The resulting product was used directly in the next step without purification.

8-Methyl-3 - ((methylamino) methyl) -2-o-tolylisoquinolin-1 (2H) -one (233 mg; 0.8 mmol) (4001) and 6-chloro-9- (tetrahydro-2H-pyran -2-yl) -9H-purine (4002) (238 mg; 1.0 mmol) was dissolved in EtOH (50 ml) and the resulting mixture was stirred at reflux for 2 hours. The mixture was allowed to cool to RT and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (2-20% MeOH / DCM), which afforded the product 8-methyl-3 - ((methyl (9- (tetrahydro-2H-pyran-2-yl) -9H-purine - 6-yl) amino) methyl) -2-o-tolylisoquinolin-1 (2H) -one (4003) (200 mg; 51% yield) as a pale yellow solid.

8-Methyl-3 - ((methyl (9- (tetrahydro-2H-pyran-2-yl) -9H-purin-6-yl) amino) methyl) -2-o-tolylisoquinolin-1 (2H) -one ( 4003) (180 mg; 0.36 mmol) was dissolved in MeOH (HCl) (50 ml) and the mixture was stirred at RT for 2 hours. An aqueous NaHCO ₃ solution was added to the reaction mixture and the pH was adjusted to 9. The mixture was filtered and the filtrate was concentrated in vacuum, which allowed to obtain the desired product 8-methyl-3 - ((methyl (9H-purin-6-yl) amino) methyl) -2-o-tolylisoquinolin-1 (2H) -one (4004) (80 mg; yield 54%) as a yellow solid.

Example 7. Synthesis of 3- (1- (9H-purin-6-ylamino) ethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one.

Scheme 18. The synthesis of 3- (1- (9H-purin-6-ylamino) ethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one (Compound 4106) is described.

To a stirred solution of 3- (hydroxymethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one 1609 (2.79 g; 10 mmol) in CH ₂ Cl ₂ (200 ml) was added MnO ₂ (5 g ) and the resulting mixture was stirred at reflux for 3 hours. The mixture was allowed to cool to RT and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (10-50% EA / PE), which afforded the product 8-methyl-1-oxo-2-o-tolyl-1,2-dihydroisoquinoline-3-carbaldehyde 4101 (2.5 g; 90% yield) as a white solid.

8-Methyl-1-oxo-2-o-tolyl-1,2-dihydroisoquinoline-3-carbaldehyde 4101 (2.4 g; 8.6 mmol) was dissolved in anhydrous THF (280 ml) and cooled to -78 ° C. in an atmosphere of nitrogen. Methyl MgBr (2 M; 5 ml; 10 mmol) was slowly added and the resulting mixture was stirred at -78 ° C for 2 hours. H ₂ O (5 ml) was added and then the solution was poured into ice-water (200 ml) and extracted with ethyl acetate (3 × 50 ml). The combined organic layer was washed with brine, dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo and the remaining product was purified by flash column chromatography on silica gel (10-50% EA / PE), which afforded the product 3- (1-hydroxyethyl) -8-methyl-2-o-tolylisoquinoline-1 (2H) -one 4102 (1.8 g; 71% yield) as a white solid.

To a solution of 3- (1-hydroxyethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one 4102 (1.6 g; 5.5 mmol) in CH ₂ Cl _{2 was} added PPh ₃ (2.88 g; 11.0 mmol) and the resulting mixture was stirred at RT for 30 minutes. Then CBr ₄ (3.64 g; 11.0 mmol) was added in portions at 0 ° C. The resulting mixture was allowed to warm to RT, stirred for 30 minutes and concentrated in vacuo. The crude product was purified by flash column chromatography on silica gel (30-50% EA / PE), which afforded the desired product 3- (1-bromoethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one 4103 ( 1.8 g; yield 91%) as a white solid.

To a stirred solution of 9- (tetrahydro-2H-pyran-2-yl) -9H-purin-6-amine 4103 (436 mg; 2 mmol) in anhydrous DMF (10 ml) was added NaH (60% in mineral oil; 77 mg; 2 mmol) and the mixture was stirred for 30 minutes. 3- (1-bromoethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one 4104 (700 mg; 2 mmol) was added. The mixture was stirred for 2 hours, poured into ice-water (200 ml) and extracted with ethyl acetate (3 × 50 ml). The combined organic layer was washed with brine (20 ml), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (10-50% MeOH / DCM), which afforded the product 8-methyl-3- (1- (9- (tetrahydro-2H-pyran-2-yl) -9H-purin-6-ylamino) ethyl) -2-o-tolylisoquinolin-1 (2H) -one 4105 (500 mg; 51% yield) as a white solid.

8-Methyl-3- (1- (9- (tetrahydro-2H-pyran-2-yl) -9H-purin-6-ylamino) ethyl) -2-o-tolylisoquinolin-1 (2H) -one 4105 (180 mg; 0.36 mmol) was dissolved in MeOH (HCl) (50 ml) and stirred for 2 hours. An aqueous solution of NaHCO _{3 was} added to the reaction mixture and the pH was adjusted to 9. The mixture was then filtered and the filtrate was concentrated in vacuo, which allowed to obtain the desired product is 3- (1- (9H-purin-6-ylamino) ethyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one 4106 (80 mg; 54% yield) as a yellow solid.

Example 8. Synthesis of 3- (4-amino-1 - ((8-methyl-1-oxo-2-o-tolyl-1,2-dihydroisoquinolin-3-yl) methyl) -1H-pyrazolo [3,4- d] pyrimidin-3-yl) -5-fluorophenyl dihydrogen phosphate.

Scheme 19. The synthesis of 3- (4-amino-1 - ((8-methyl-1-oxo-2-o-tolyl-1,2-dihydroisoquinolin-3-yl) methyl) -1H-pyrazolo is described [3,4 -d] pyrimidin-3-yl) -5-fluorophenyl dihydrogen phosphate (Compound 4303).

3 - ((4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d] pyrimidin-1-yl) methyl) -8-methyl-2-o-tolylisoquinolin-1 (2H) -one 4301 (250 mg; 0.5 mmol) was dissolved in anhydrous THF (15 ml) in the dark in a round bottom flask (coated with aluminum foil) and cooled to 0 ° C in argon atmosphere. CBr ₄ (498 mg; 1.5 mmol) was added, followed by diethyl phosphite (129 μl; 1.0 mmol) and triethylamine (417 μl; 1.5 mmol). The resulting mixture was stirred in the dark at a temperature of 0 ° C to CT for 16 hours. The mixture was then partitioned between ethyl acetate and brine. The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography, eluting with methanol and dichloromethane, which afforded the desired product 3- (4-amino-1 - ((8-methyl-1-oxo-2-o-tolyl-1,2-dihydroisoquinolin-3- il) methyl) -1H-pyrazolo [3,4-d] pyrimidin-3-yl) -5-fluorophenyl diethylphosphate 4302 (200 mg; yield 62%) as an off-white solid.

3- (4-amino-1 - ((8-methyl-1-oxo-2-o-tolyl-1,2-dihydroisoquinolin-3-yl) methyl) -1H-pyrazolo [3,4-d] pyrimidine- 3-yl) -5-fluorophenyl diethyl phosphate 4302 (170 mg; 0.26 mmol) was dissolved in anhydrous CH ₃ CN (5 ml) and cooled to 0 ° C under argon. TMSBr (trimethylsilyl-Br) (0.34 ml; 2.64 mmol) was slowly added via syringe and the resulting mixture was stirred at 0 ° C to CT for 16 hours. LC-MS (liquid chromatography / mass spectrometry) data ) showed that a small amount of the starting material was consumed, therefore, additional amounts of TMSBr (0.1 ml) were added and stirred at RT for 5 hours. LC-MS data showed complete conversion of the starting material. The mixture was concentrated in vacuo and the residue was dissolved in Et ₂ O (10 ml) and H ₂ O (0.5 ml) and stirred for 30 minutes. The mixture was concentrated in vacuo to afford the desired product 3- (4-amino-1 - ((8-methyl-1-oxo-2-o-tolyl-1,2-dihydroisoquinolin-3-yl) methyl) -1H- pyrazolo [3,4-d] pyrimidin-3-yl) -5-fluorophenyl dihydrogen phosphate 4303 (140 mg; 91% yield).

Example 9. IC ₅₀ Values for Individual Compounds

Table 3 In vitro IC ₅₀ data for individual compounds IC ₅₀ (nM) + (greater than 10 μM) ++ (less than 10 μm) +++ (less than 1 μM) ++++ (less than 100 nM) PI3K δ Compound No. Compound No. Compound No. Compound No. 1, 5, 22, 27, 38, 39, 40, 41, 46, 92, 117, 118, 120, 129, 132, 164, 165, 172, 188, 186, 193, 194 4, 14, 15, 17, 18, 21, 26, 29, 31, 32, 34, 35, 36, 42, 43, 44, 45, 47, 49, 57, 69, 71, 85, 87, 94, 106, 107, 143, 175, 179, 181, 182, 183, 187, 189, 192 2, 3, 6, 7, 8, 9, 10, 11, 12, 13, 16, 19, 20, 23, 24, 25, 28, 30, 33, 37, 48, 50, 51, 52, 53, 54, 55, 56, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 70, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 86, 88, 89, 90, 91, 93, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 108, 109, 110, 111, 112, 113, 114, 115, 119, 123, 124, 125, 126, 128, 134, 135, 136, 137, 138, 139, 141, 142, 144, 145, 146, 147, 148, 149, 150, 151. 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 166, 167, 168, 169, 170, 171, 173, 174, 176, 177 178, 180, 185, 188, 190, 191 PI3K γ Compound No. Compound No. Compound No. Compound No. 1, 4, 5, 18, 38, 43, 60, 69, 169, 172, 192, 193, 194 17, 34, 35, 37, 38, 40, 42, 57, 61, 65, 91, 92, 94, 105, 107, 140, 164, 170, 175, 179, 184, 186, 187, 189 2, 8, 9, 10, 11, 14, 15, 20, 22, 27, 28, 39, 41, 46, 47, 49, 51, 55, 58, 66, 70, 71, 73, 76, 78, 80, 93, 98, 99, 100, 103, 104, 106, 108, 109, 161, 162, 163, 165, 166, 180, 188 3, 6, 7, 12, 13, 16, 19, 21, 23, 24, 25, 26, 29, 30, 31, 33, 36, 44, 45, 48, 50, 52, 53, 54, 56, 59, 62, 63, 64, 67, 68, 72, 74, 75, 77, 79, 81, 82, 83, 84, 86, 87, 88, 89, 90, 95, 96, 97, 101, 102, 142, 145,
146, 147, 148, 149, 150, 151, 152, 160, 167, 168, 171, 173, 174, 176, 177, 178, 182, 185, 190, 191 PI3K α Compound No. Compound No. Compound No. Compound No. 6, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 54, 55, 56, 57, 58, 59, 60, 61, 62, 64, 65, 67, 68, 69, 70, 71, 72, 73, 74, 79, 80, 81, 82, 83, 85, 87, 88, 91, 93, 96, 98, 99, 100, 103, 104, 105, 106, 107, 109, 110, 111, 112, 114, 140, 146, 160, 161, 162, 163, 164, 165, 166, 167, 169, 170, 172, 174, 175, 179, 180, 183, 184, 188, 190, 191, 192, 193, 194 3, 7, 63, 66, 84, 86, 89, 90, 97, 108, 113, 115, 152, 168, 171, 173, 185, 190 53, 95, 101, 102, 145, 147, 149, 151, 177 142, 148, 150, 153, 154, 155, 156, 157, 158, 159, 176 PI3K β Compound No.
8, 9, 10, 11, 14, 21, 22, 24, 26, 27, 28, 29, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 46, 52, 54, 56, 57, 59, 60, 64, 68, 69, 70, 73, 76, 78, 79, 80, 87, 88, 91, 93, 98, 103, 104, 105, 107, 109, 112, 140, 146, 152, 162, 163, 164, 165, 166, 169, 170, 172, 175, 179, 180, 183, 184, 188, 190, 191, 192, 193, 194 Compound No.
3, 12, 13, 23, 25, 53, 55, 58, 61, 63, 65, 67, 71, 72, 74, 75, 77, 81, 82, 83, 84, 85, 86, 96, 99, 106, 108, 110, 111, 113, 114, 115, 145, 147, 149, 151, 154, 158, 160, 161, 167, 168, 171, 173, 174, 177, 178,190, 191 Compound No.
7, 62, 66, 82, 89, 90, 95, 97, 100, 102, 150, 153, 159, 176, 185 Compound No.
101, 142, 155, 156, 157 B cell proliferation, EC ₅₀ (nM) Compound No. Compound No. Compound No. Compound No. 38, 162 1, 2, 5, 22, 26, 27, 39, 40, 43, 49, 57, 71, 87, 112 4, 8, 9, 10, 11, 14, 15, 18, 19, 20, 21, 24, 25, 28, 29, 30, 31, 32, 34, 35, 36, 41, 42, 45, 46, 47, 50, 51, 61, 69, 70, 76, 77, 78, 79, 80, 85, 86, 91, 98, 100, 103, 104, 105, 106, 107, 110, 111, 114, 119, 124, 133, 135, 145, 152, 161, 162, 163, 169 3, 6, 7, 12, 13, 16, 17, 23, 33, 37, 44, 48, 53, 54, 55, 62, 63, 66, 67, 68, 72, 73, 74, 75, 81, 82, 83, 84, 88, 89, 90, 93, 95, 96, 97, 99, 101, 102, 108, 109, 113, 115, 123, 125, 126, 128, 134, 136, 137, 138, 139, 141, 142, 144, 146, 147, 148, 149, 150, 151, 153, 154, 155, 156, 157, 158, 159,
160, 166, 167, 168, 170, 171, 173, 174, 176, 177, 178, 180, 187, 185, 188, 190, 191

Table 4. The structures of the compounds giving the IC ₅₀ are shown in Table 3

Structure

Compound 1 Compound 2 Compound 3 Compound 4 Compound 5

Compound 6 Compound 7 Compound 8 Compound 9 Compound 10

Compound 11 Compound 12 Compound 13 Compound 14 Compound 15

Structure

Compound 16 Compound 17 Compound 18 Compound 19 Compound 20

Compound 21 Compound 22 Compound 23 Compound 24 Compound 25

Compound 26 Compound 27 Compound 28 Compound 29 Compound 30

Compound 31 Compound 32 Compound 33 Compound 34 Compound 35

Compound 36 Compound 37 Compound 38 Compound 39 Compound 40

Structure

Compound 41 Compound 42 Compound 43 Compound 44 Compound 45

Compound 46 Compound 47 Compound 48 Compound 49 Compound 50

Compound 51 Compound 52 Compound 53 Compound 54 Compound 55

Compound 56 Compound 57 Compound 58 Compound 59 Compound 60

Compound 61 Compound 62 Compound 63 Compound 64 Compound 65

Structure

Compound 66 Compound 67 Compound 68 Compound 69 Compound 70

Compound 71 Compound 72 Compound 73 Compound 74 Compound 75

Compound 76 Compound 77 Compound 78 Compound 79 Compound 80

Compound 81 Compound 82 Compound 83 Compound 84 Compound 85

Compound 86 Compound 87 Compound 88 Compound 89 Compound 90

Structure

Compound 91 Compound 92 Compound 93 Compound 94 Compound 95

Compound 96 Compound 97 Compound 98 Compound 99 Compound 100

Compound 101 Compound 102 Compound 103 Compound 104 Compound 105

Compound 106 Compound 107 Compound 108 Compound 109 Compound 110

Compound 111 Compound 112 Compound 113 Compound 114 Compound 115

Structure

Compound 116 Compound 117 Compound 118 Compound 119 Compound 120

Compound 121 Compound 122 Compound 123 Compound 124 Compound 125

Compound 126 Compound 127 Compound 128 Compound 129 Compound 130

Compound 131 Compound 132 Compound 133 Compound 134 Compound 135

Compound 136 Compound 137 Compound 138 Compound 139 Compound 140

Structure

Compound 141 Compound 142 Compound 143 Compound 144 Compound 145

Compound 146 Compound 147 Compound 148 Compound 149 Compound 150

Compound 151 Compound 152 Compound 153 Compound 154 Compound 155

Compound 156 Compound 157 Compound 158 Compound 159 Compound 160

Structure

Compound 161 Compound 162 Compound 163 Compound 164 Compound 165

Compound 166 Compound 167 Compound 168 Compound 169 Compound 170

Compound 171 Compound 172 Compound 173 Compound 174 Compound 175

Compound 176 Compound 177 Compound 178 Compound 179 Compound 180

Compound 181 Compound 182 Compound 183 Compound 184 Compound 185

Structure

Compound 186 Compound 187 Compound 188 Compound 189 Compound 190

Compound 191 Compound 192 Compound 193 Compound 194

Example 10. Analysis of expression and inhibition of p110a / p85a, p110β / p85α, p110δ / p85α and p110γ.

Class I PI3-K can either be acquired (p110α / p85α, p110β / p85α, p110δ / p85α from Upstate and p110γ from Sigma), or expressed as previously described (Knight et al., 2004). IC50 values are measured using either a standard TLC assay for lipid kinase activity (described below) or a high throughput membrane capture assay. Kinase reactions are carried out by preparing a reaction mixture containing a kinase, an inhibitor (final concentration of DMSO 2%), a buffer (25 mM HEPES; pH 7.4; 10 mM MgCl ₂ ) and freshly prepared, sonicated phosphatidylinositol (100 μg / ml). Reactions are initiated by adding an ATP solution containing 10 μCi γ- ³² P-ATP to a final concentration of 10 or 100 μM and allowed to proceed for 5 minutes at room temperature. Next, to conduct TLC analysis of the reaction, stop adding 105 μl of 1 N. HCl, then 160 μl of a mixture of CHCl ₃ : MeOH (1: 1). The biphasic mixture is vortexed, centrifuged quickly and the organic phase is transferred to a new tube using a pipette tip for application onto a gel precoated with CHCl ₃ . This extract is spotted on TLC plates and chromatographed for 3-4 hours in a solution of 65:35 n-propanol: 1 M acetic acid. Then TLC plates are dried, placed on the screen of the phospho-imager (Storm, Amersham) and quantified. The kinase activity for each compound is measured for 10-12 inhibitor concentrations, representing two-fold dilutions from the highest concentration tested (usually 200 μM). For compounds exhibiting significant activity, IC50 determinations are repeated two to four times, and the values presented are the average of these independent measurements.

Other commercially available PI3-K activity assay kits or systems are available. Commercially available kits or systems can be used to screen PI3-K inhibitors and / or agonists, including, but not limited to, PI3 kinases α, β, δ, and γ. A typical system is Upstate's PI3 kinase (human) assay system. This analysis can be carried out in accordance with the methods proposed by the manufacturer. Briefly, this analysis is a time-resolved fluorescence resonance energy transfer study (TR-FRET) in which the PIP3 product resulting from the action of PI3-K is indirectly measured. The kinase reaction is carried out in a microtiter plate (for example, a 384-well microtiter plate). The total volume of the reaction mixture is approximately 20 μl per well. In the first stage, 2 μl of a solution of the test compound in 20% dimethyl sulfoxide is placed in each well, which results in a final concentration of DMSO of 2%. Then, approximately 14.5 μl of a kinase / PIP2 (phosphatidylinositol diphosphate) mixture (diluted in 1X reaction buffer) was added to each well to obtain a final kinase concentration of 0.25-0.3 μg / ml and PIP2 10 μM. The tablet is sealed and incubated for 15 minutes at room temperature. To start the reaction, 3.5 μl of ATP solution (diluted in 1X reaction buffer) was added to each well until a final ATP concentration of 10 μM was obtained. The tablet is sealed and incubated for 1 hour at room temperature. The reaction is stopped by adding 5 μl of the solution to stop the reaction per well and then 5 μl of the detection mixture per well. The tablet is sealed, incubated for 1 hour at room temperature and then read on an appropriate tablet reader. Data is analyzed and IC50 values are obtained using GraphPad Prism 5.

Example 11. Analysis of expression and inhibition of Al

Cross activity or lack thereof for one or more of the compounds of the present invention with respect to Al kinase can be measured in accordance with any method known in the art or with the methods set forth below. The compounds set forth in this application can be assayed in triplicate for the recombinant full length Al or Al (T315I) (Upstate) in an assay containing 25 mM HEPES, pH 7.4; 10 mM MgCl ₂ ; 200 μM ATP (2.5 μCi γ- ³² P-ATP) and 0.5 mg / ml BSA (bovine serum albumin). The optimized peptide substrate for Al EAIYAAPFAKKK was used as a phospho acceptor (200 μM). The reactions are stopped by applying the reaction mixture in the form of spots on phosphocellulose plates, which are washed with 0.5% phosphoric acid (approximately 6 times, each time for 5-10 minutes). The plates are dried and the transferred radioactive compound is quantified by imaging using phosphorimaging radionuclide.

Example 12. Analysis of the expression and inhibition of Hc

Cross-activity or lack thereof for one or more compounds of the present invention with respect to the Hck kinase can be measured in accordance with any method known in the art or with the methods set forth below. The compounds set forth in this application can be assayed in triplicate for a recombinant full-length HSC in an assay containing 25 mM HEPES, pH 7.4; 10 mM MgCl ₂ ; 200 μM ATP (2.5 μCi γ- ³² P-ATP) and 0.5 mg / ml BSA. The optimized peptide substrate for the EIYGEFKKK Src family kinases is used as a phospho acceptor (200 μM). The reactions are stopped by applying the reaction mixture in the form of spots on phosphocellulose plates, which are washed with 0.5% phosphoric acid (approximately 6 times, each time for 5-10 minutes). The plates are dried and the radioactive compound is quantified by imaging using a phosphorus radionuclide.

Example 13. Analysis of expression and inhibition of the insulin receptor (IR)

Cross activity or lack thereof for one or more compounds of the present invention with respect to the IR receptor kinase can be measured in accordance with any method known in the art or with the methods set forth below. The compounds described herein can be assayed in triplicate for the recombinant kinase domain of the insulin receptor (Upstate) in an assay containing 25 mM HEPES, pH 7.4; 10 mM MgCl ₂ ; 10 mM MnCl ₂ ; 200 μM ATP (2.5 μCi γ- ³² P-ATP) and 0.5 mg / ml BSA. Poly-EY (a copolymer of glutamic acid and tyrosine) (Sigma; 2 mg / ml) is used as a substrate. The reaction is stopped by applying the reaction mixture in the form of spots on nitrocellulose, which is washed with a mixture of 1 M MaCl / 1% phosphoric acid (approximately 6 times, each time for 5-10 minutes). The plates are dried and the radioactive compound is quantified by imaging using a phosphorus radionuclide.

Example 14. Src expression and inhibition assays

Cross-activity or lack thereof for one or more compounds of the present invention in relation to Src kinase can be measured in accordance with any method known in the art, or with the methods described below. The compounds described herein can be assayed in triplicate for recombinant full length Src or Src (T338I) in an assay containing 25 mM HEPES, pH 7.4; 10 mM MgCl ₂ ; 200 μM ATP (2.5 μCi γ- ³² P-ATP) and 0.5 mg / ml BSA. The optimized peptide substrate for the EIYGEFKKK Src family kinases is used as a phospho acceptor (200 μM). The reactions are stopped by applying the reaction mixture in the form of spots on phosphocellulose plates, which are washed with 0.5% phosphoric acid (approximately 6 times, each time for 5-10 minutes). The plates are dried and the radioactive compound is quantified by imaging using a phosphorus radionuclide.

Example 15. DNA-PK Expression and Inhibition Assays (DNAK)

Cross-activity or lack thereof for one or more compounds of the present invention with respect to DNAK kinase can be measured in accordance with any technique known in the art. DNA-PK can be purchased from Promega and analyzed using the DNA-PK analysis system (Promega) in accordance with the manufacturer's instructions.

Example 16. Analysis of expression and inhibition of mTOR

Cross-activity or lack thereof for one or more compounds of the present invention with respect to mTOR can be measured in accordance with any method known in the art or with the methods described below. Compounds described in this application can be tested for recombinant mTOR (Invitrogen) in an assay containing 50 mM HEPES, pH 7.5; 1 mM EGTA (ethylene glycol tetraacetic acid); 10 mM MgCl ₂ ; 2.5 mM, 0.01% Tween; 10 μM ATP (2.5 μCi μ- ³² P-ATP) and 3 μg / ml BSA. Recombinant PHAS-1 / 4EBP1 (phosphorylated heat and acid stable protein-1 / 4E-binding protein-1 (phosphorylated heat and acid stable protein-1 / 4E binding protein-1)) rats (Calbiochem, 2 mg / ml). The reaction is stopped by applying the reaction mixture in the form of spots on nitrocellulose, which is washed with a mixture of 1 M MaCl / 1% phosphoric acid (approximately 6 times, each time for 5-10 minutes). The plates are dried and the radioactive compound is quantified by imaging using a phosphorus radionuclide.

Other kits or systems for mTOR activity analysis are commercially available. For example, you can use the LanthaScreen ™ kinase assay kit (Invitrogen) to test the mTOR inhibitors described in this application. This analysis is based on the use of a time-resolved FRET platform where the 4EBP1 phosphorylation labeled with GFP (green fluorescent protein) is measured by mTOR kinase. The kinase reaction is carried out in a white 384-well microtiter plate. The total reaction volume is 20 μl per well, and the reaction buffer has the following composition: 50 mM HEPES, pH 7.5; 0.01% polysorbate 20; 1 mM EGTA; 10 mM MnCl ₂ and 2 mM DTT. In the first stage, 2 μl of a solution of the test compound in 20% dimethyl sulfoxide is placed in each well, which results in a final concentration of DMSO of 2%. Then add 8 μl per well of mTOR diluted in the reaction buffer to a final concentration of 60 ng / ml. To start the reaction, 10 μl of an ATP / GFP-4EBP1 mixture (diluted in reaction buffer) was added to each well to obtain a final ATP concentration of 10 μM and 0.5 μM GFP-4EBP1. The tablet is sealed and incubated for 1 hour at room temperature. The reaction is stopped by adding 10 μl per well of a mixture of Tb-anti-pT46-4EBP-antibody / EOTA (ethylenediaminetetraacetic acid) (diluted in TR-FRET buffer) to obtain a final antibody concentration of 1.3 nM and 6.7 mM EDTA . The plate is sealed, incubated for 1 hour at room temperature and then read on a tablet reader with the settings for TR-FRET LanthaScreen ™. Data is analyzed and IC50 values are obtained using GraphPad Prism 5.

Example 17. Analyzes of expression and inhibition of vascular endothelial growth factor receptor.

Cross-activity or lack thereof for one or more compounds of the present invention with respect to the VEGF receptor can be measured in accordance with any method known in the art, or with the methods set forth below. The compounds described herein can be tested against the kinase domain of the recombinant KDR (Invitrogen kinase insertion receptor kinase) receptor in an assay containing 25 mM HEPES, pH 7.4; 10 mM MgCl ₂ ; 0.1% BME (2 -mercaptoethanol); 10 μM ATP (2.5 μCi γ- ³² P-ATP) and 3 μg / ml BSA. Poly-EY (Sigma; 2 mg / ml) was used as a substrate. The reactions were stopped by applying the reaction mixture as spots on nitrocellulose, which is washed with a mixture of 1 M NaCl / 1% phosphoric acid (approximately 6 times, each time for 5-10 minutes). active compound quantitatively measured by visualization using phosphorus radionuclide.

Example 18. Analyzes of the expression and inhibition of the E4 receptor B4 (EpnB4)

Cross-activity or lack thereof for one or more compounds of the present invention with respect to EphB4 can be measured in accordance with any method known in the art, or with the methods set forth below. The compounds described in this application can be tested against the kinase domain of the recombinant E4 receptor B4 (Invitrogen) in an assay containing 25 mM HEPES, pH 7.4; 10 mM MgCl ₂ ; 0.1% BME; 10 μM ATP (2.5 μCi μ- ³² P-ATP) and 3 μg / ml BSA. Poly-EY (Sigma; 2 mg / ml) was used as a substrate. The reactions are stopped by applying the reaction mixture in the form of spots on nitrocellulose, which is washed with a mixture of 1 M NaCl / 1% phosphoric acid (approximately 6 times, each time for 5-10 minutes). The plates are dried and the radioactive compound is quantified by imaging using a phosphorus radionuclide.

Example 19. Analysis of expression and inhibition of the epidermal growth factor receptor (EGFR)

Cross-activity or lack thereof for one or more of the compounds of the present invention with respect to EGFR kinase can be measured in accordance with any method known in the art or with the methods set forth below. The compounds described in this application can be tested against the kinase domain of the recombinant EGF receptor (Invitrogen) in an assay containing 25 mM HEPES, pH 7.4; 10 mM MgCl ₂ ; 0.1% BME; 10 μM ATP (2.5 μCi μ- ³² P-ATP) and 3 μg / ml BSA. Poly-EY (Sigma; 2 mg / ml) was used as a substrate. The reactions are stopped by applying the reaction mixture in the form of spots on nitrocellulose, which is washed with a mixture of 1 M NaCl / 1% phosphoric acid (approximately 6 times, each time for 5-10 minutes). The plates are dried and the radioactive compound is quantified by imaging using a phosphorus radionuclide.

Example 20. KIT expression and inhibition assays

Cross-activity or lack thereof for one or more compounds of the present invention with respect to KIT kinase can be measured in accordance with any method known in the art or with the methods set forth below. The compounds described in this application can be tested against the kinase domain of the recombinant KIT (Invitrogen) in an assay containing 25 mM HEPES, pH 7.4; 10 mM MgCl ₂ ; 1 mM DTT; 10 mM MnCl ₂ ; 10 μM ATP (2.5 μCi μ- ³² P-ATP) and 3 μg / ml BSA. Poly-EY (Sigma; 2 mg / ml) was used as a substrate. The reactions are stopped by applying the reaction mixture in the form of spots on nitrocellulose, which is washed with a mixture of 1 M NaCl / 1% phosphoric acid (approximately 6 times, each time for 5-10 minutes). The plates are dried and the radioactive compound is quantified by imaging using a phosphorus radionuclide.

Example 21. Analysis of expression and inhibition of RET

Cross activity or lack thereof for one or more of the compounds of the present invention with respect to RET kinase can be measured in accordance with any method known in the art or with the methods set forth below. The compounds described in this application can be tested against the kinase domain of recombinant RET (Invitrogen) in an assay containing 25 mM HEPES, pH 7.4; 10 mM MgCl ₂ ; 2.5 mM DTT; 10 μM ATP (2.5 μCi μ- ³² P-ATP) and 3 μg / ml BSA. The optimized peptide substrate for Abl EAIYAAPFAKKK is used as a phospho acceptor (200 μM). The reactions are stopped by applying the reaction mixture in the form of spots on phosphocellulose plates, which are washed with 0.5% phosphoric acid (approximately 6 times, each time for 5-10 minutes). The plates are dried and the radioactive compound is quantified by imaging using a phosphorus radionuclide.

Example 22. Analysis of expression and inhibition of platelet growth factor receptor (PDGFR)

Cross-activity or lack thereof for one or more of the compounds of the present invention with respect to PDGFR kinase can be measured in accordance with any method known in the art or with the methods set forth below. The compounds described in this application can be tested against the kinase domain of the recombinant PDGF receptor (Invitrogen) in an assay containing 25 mM HEPES, pH 7.4; 10 mM MgCl ₂ ; 2.5 mM DTT; 10 μM ATP (2.5 μCi μ- ³² P-ATP) and 3 μg / ml BSA. The optimized peptide substrate for Abl EAIYAAPFAKKK is used as a phospho acceptor (200 μM). The reactions are stopped by applying the reaction mixture in the form of spots on phosphocellulose plates, which are washed with 0.5% phosphoric acid (approximately 6 times, each time for 5-10 minutes). The plates are dried and the radioactive compound is quantified by imaging using a phosphorus radionuclide.

Example 23. Analysis of the expression and inhibition of Fms-like tyrosine kinase 3 (FLT-3)

Cross-activity or lack thereof for one or more compounds of the present invention with respect to FLT-3 kinase can be measured in accordance with any method known in the art or with the methods set forth below. The compounds described in this application can be tested against the kinase domain of recombinant FLT-3 (Invitrogen) in an assay containing 25 mM HEPES, pH 7.4; 10 mM MgCl ₂ ; 2.5 mM DTT; 10 μM ATP (2.5 μCi μ- ³² P-ATP) and 3 μg / ml BSA. The optimized peptide substrate for Abl EAIYAAPFAKKK is used as a phospho acceptor (200 μM). The reactions are stopped by applying the reaction mixture in the form of spots on phosphocellulose plates, which are washed with 0.5% phosphoric acid (approximately 6 times, each time for 5-10 minutes). The plates are dried and the transferred radioactive compound is quantified by imaging using a phosphorus radionuclide.

Example 24. Analysis of expression and inhibition of receptor tyrosine kinase TEK (TIE2)

Cross-activity or lack thereof for one or more compounds of the present invention with respect to TIE2 kinase can be measured in accordance with any method known in the art or with the methods set forth below. The compounds described in this application can be tested against the kinase domain of recombinant TIE2 (Invitrogen) in an assay containing 25 mM HEPES, pH 7.4; 10 mM MgCl ₂ ; 2 mM DTT; 10 mM MnCl ₂ ; 10 μM ATP (2.5 μCi μ- ³² P-ATP) and 3 μg / ml BSA. Poly-EY (Sigma; 2 mg / ml) was used as a substrate. The reactions are stopped by applying the reaction mixture in the form of spots on nitrocellulose, which is washed with a mixture of 1 M NaCl / 1% phosphoric acid (approximately 6 times, each time for 5-10 minutes). The plates are dried and the transferred radioactive compound is quantified by imaging using a phosphorus radionuclide.

Example 25. Analysis of the activation and proliferation of b-cells

The ability of one or more compounds of the invention to inhibit the activation and proliferation of B cells is determined in accordance with standard techniques known in the art. For example, an in vitro method of cell proliferation analysis has been established in which the metabolic activity of living cells is measured. The analysis was carried out in a 96-well microtiter plate using Alamar Blue dye reduction. Balb / c spleen B cells were purified using a Ficoll-Paque ™ PLUS gradient, followed by magnetic cell separation using a MACS B cell isolation kit (Miletenyi). Cells are seeded onto plates in a volume of 90 μl in an amount of 50,000 cells / well in B-cell medium (RPMI medium (Roswell Park Memorial Institute) + 10% FBS (fetal bovine serum) + penicillin / streptomycin + 50 μM bME) + 5 mM HEPES). The compound set forth in this application is diluted in B cell medium and added in a volume of 10 μl. The plates are incubated for 30 min at 37 ° C and 5% CO ₂ (final concentration of DMSO 0.2%). Then, 50 μl of a mixture for stimulating B cells containing either LPS (lipopolysaccharide) (10 μg / ml) or F (ab ') ₂ donkey antibodies against mouse IgM (5 μg / ml) plus recombinant IL4 (interleukin-4) is added ) mice (2 ng / ml) in B cell medium. The plates are incubated for 72 hours at 37 ° C and 5% CO ₂ . Alamar Blue reagent was added to each well in a volume of 15 μl and the plates were incubated for 5 hours at 37 ° C and 5% CO ₂ . Alamar Blue fluorescence is read at 560Ex / 590Et (excitation at 560 nm, emission at 590 nm) and IC50 or EC50 values are calculated using GraphPad Prism 5.

Example 26. Analysis of tumor cell line proliferation

The ability of one or more compounds of the invention to inhibit the proliferation of a tumor cell line is determined in accordance with standard techniques known in the art. For example, an in vitro cell proliferation assay can be performed in which the metabolic activity of living cells is measured. This assay is performed in a 96-well microtiter plate using Alamar Blue reduction. Human tumor cell lines are obtained from ATCC (American Type Culture Collection) (for example, MCF7, U-87 MG; MDA-MB-468, PC-3), grown to confluency in T75 bottles, trypsinization using 0.25% trypsin, washed once with tumor cell medium (DMEM (Dulbecco's modified Eagle's medium) + 10% FBS) and plated on plates in a volume of 90 μl in an amount of 5000 cells / well in the tumor cell medium. The compound described in this application is diluted in a medium for tumor cells and added in a volume of 10 μl. The plates are incubated for 72 hours at 37 ° C and 5% CO ₂ . Alamar Blue reagent was added to each well in a volume of 15 μl and the plates were incubated for 3 hours at 37 ° C and 5% CO ₂ . Alamar Blue fluorescence is read at 560Ex / 590 Em and IC50 values are calculated using GraphPad Prism 5.

Example 27. Antitumor activity in vivo

You can evaluate the compounds described in this application, in a number of models of human and mouse tumors.

Models of tumors resistant to paclitaxel.

1. The model of ovarian cancer obtained on clinical material.

This tumor model was created as a result of a biopsy of a tumor of a patient with ovarian cancer. Tumor biopsy taken from the patient.

The compounds described in this application are administered to athymic mice bearing tumors with an established stage of development using the scheme: every 2 days × 5.

2. A2780Tax human ovarian cancer xenograft (with mutant tubulin).

A2780Tax is a model of human ovarian cancer resistant to paclitaxel. It is obtained from the sensitive A2780 baseline as a result of co-incubation with paclitaxel and verapamil, an agent that eliminates MDR (multidrug resistance). It was shown that the mechanism of resistance to it is not related to the MDR mechanism and is explained by the presence of a mutation in the gene encoding the beta-tubulin protein.

The compounds described in this application can be administered to mice bearing tumors with an established stage of development, according to the scheme: every 2 days × 5.

3. HCT116 / VM46 human colon cancer xenograft (multidrug resistant)

HCT116 / VM46 is an MDR-resistant colon cancer from the sensitive HCT116 baseline. With in vivo growth in athymic mice, HCT116 / VM46 invariably shows high resistance to paclitaxel.

The compounds described in this application can be administered to mice bearing tumors with an established stage of development, according to the scheme: every 2 days × 5.

4. Model of mouse sarcoma M5076

M5076 is a mouse fibrosarcoma that is naturally resistant to paclitaxel in vivo.

The compounds described in this application can be administered to mice bearing tumors with an established stage of development, according to the scheme: every 2 days × 5.

One or more of the compounds of the invention can be used in combination with other in vivo therapeutic agents for HCT / VM46 multidrug-resistant human colon cancer xenografts or for any other model known in the art, including those described herein.

Example 28. Stability analysis in the presence of microsomes

The stability of one or more of the compounds of the invention is determined in accordance with standard procedures known in the art. For example, the stability of one or more compounds of the invention is determined in an in vitro assay. In particular, a stability analysis can be performed in the presence of microsomes in vitro, in which the stability of one or more compounds of the invention is measured when interacting with microsomes from mouse, rat, or human liver. The interaction of microsomes with compounds is carried out in an Eppendorf tube with a capacity of 1.5 ml. Each tube contains 0.1 μl of NADPH (nicotinamide adenine dinucleotide reduced) (10.0 mg / ml); 75 μl of microsomes from the liver of a mouse, rat, or human (20.0 mg / ml); 0.4 μl of 0.2 M phosphate buffer and 425 μl of ddH ₂ O (twice distilled water). A negative control tube (without NADPH) contains 75 μl of microsomes from mouse, rat, or human liver (20.0 mg / ml); 0.4 μl of 0.2 M phosphate buffer and 525 μl of ddH ₂ O. The reaction is initiated by adding 1.0 μl of 10.0 mm test compound. Test tubes with the reaction mixture are incubated at 37 ° C. 100 μl of sample is taken into a new Eppendorf tube containing 300 μl of cold methanol, after 0, 5, 10, 15, 30 and 60 minutes from the start of the reaction. Samples were centrifuged at 15,000 rpm to remove protein. The supernatant of the centrifuged sample is transferred to a new tube. The concentration of the stable compound in the supernatant after interaction with microsomes was measured by liquid chromatography / mass spectrometry (LC-MS).

Example 29. Analysis of stability in blood plasma

The stability of one or more compounds of the invention in blood plasma is determined in accordance with standard techniques known in the art. See, for example, Rapid Commun. Mass Spectrom., 10: 1019-1026. The following procedure is an HPLC-MS / MS analysis using human blood plasma; other types of plasma are also possible, including monkey, dog, rat and mouse. Frozen heparinized human blood plasma is thawed in a cold water bath and centrifuged for 10 minutes at 2000 rpm at 4 ° C before use. The compound of the invention is added from a 400 μM concentrated solution to an aliquot of preheated plasma to obtain a final volume for analysis of 400 μl (or 800 μl to determine the half-life) containing 5 μM of test compound and 0.5% DMSO. The reaction mixtures are incubated with shaking for 0 minutes and 60 minutes at 37 ° C or for 0, 15, 30, 45 and 60 minutes at 37 ° C to determine the half-life. The reactions are stopped by transferring 50 μl of the incubation mixture to 200 μl of ice-cold acetonitrile and stirring by shaking for 5 minutes. Samples were centrifuged at 6000 xg for 15 minutes at 4 ° C and 120 μl of the supernatant were taken into clean tubes. The samples were then evaporated to dryness and analyzed using HPLC-MS / MS.

Where desired, at the same time as the test compounds, one or more control compounds or comparison compounds (5 μM) are tested: one compound, propoxicaine, with low plasma stability, and another compound, propantheline, with intermediate plasma stability.

Samples are re-diluted in acetonitrile / methanol / water (1/1/2, v / v / v) and analyzed using reverse phase (RP) HPLC-MS / MS using a selective reaction monitoring method (SRM) . HPLC conditions include a dual LC pump with auto-dispenser, combined operation, C12, 2 × 20 mm column and gradient program. Peak areas corresponding to analytes are recorded using HPLC-MS / MS. The ratio of the starting compound remaining after 60 minutes and the amount at time zero, expressed as a percentage, is taken as plasma stability. In the case of determining the half-life, it is determined from the slope of the initial linear portion of the logarithmic curve of the dependence of the remaining compound (%) on time, assuming the presence of first-order kinetics.

Example 30. Chemical stability

The chemical stability of one or more of the compounds of the invention is determined in accordance with standard procedures known in the art. The following are details of a typical method for determining the chemical stability of a compound of the invention. The standard buffer used for chemical stability analysis is phosphate buffered saline (PBS) with a pH of 7.4; other suitable buffers may be used. The compound of the invention is added from a 100 μM concentrated solution to an aliquot of PBS (in duplicate) to obtain a final volume for analysis of 400 μl containing 5 μM test compound and 1% DMSO (a sample with a total volume of 700 μl is prepared to determine the half-life ) The reaction mixtures were incubated with shaking for 0 minutes and 24 hours at 37 ° C; to determine the half-life, the samples are incubated for 0, 2, 4, 6 and 24 hours. Reactions are stopped by immediately adding 100 μl of the incubation mixture to 100 μl of acetonitrile and vortexing for 5 minutes. Samples are further stored at −20 ° C. until HPLC-MS / MS analysis is performed. Where desired, a control or reference compound, such as chlorambucil (5 μM), is tested simultaneously with the compound of interest of the invention since this compound is substantially hydrolyzed within 24 hours. Samples were analyzed using (RP) HPLC-MS / MS using a selective reaction monitoring (SRM) method. HPLC conditions include a dual LC pump with auto-dispenser, combined operation, C12, 2 × 20 mm column and gradient program. Peak areas corresponding to analytes are recorded using HPLC-MS / MS. The ratio of the starting compound remaining after 24 hours and the amount at time zero, expressed as a percentage, is taken as chemical stability. In the case of determining the half-life, it is determined from the slope of the initial linear portion of the logarithmic curve of the dependence of the remaining compound (%) on time, assuming the presence of first-order kinetics.

Example 31. Analysis of Akt kinase

Cells containing components of the Akt / mTOR pathway, including, but not limited to, L6 myoblasts, B-ALL cells, B cells, T cells, leukemia cells, bone marrow cells, p190 transduced cells, cells with a Philadelphia positive chromosome ( Ph +) and mouse embryonic fibroblasts are usually grown in cell growth media such as DMEM supplemented with fetal bovine serum and / or antibiotics and grown to a confluent state.

In order to compare the effect of one or more of the compounds described in this application on Akt activation, these cells are depleted in serum overnight and incubated with one or more of the compounds described in this application, or with about 0.1% DMSO in for about 1 minute to about 1 hour, after which they are stimulated with insulin (e.g. 100 nM) for about 1 minute to about 1 hour. Cells are lysed by scraping them into ice-cold lysis buffer containing detergents such as sodium dodecyl sulfate and protease inhibitors (e.g. PMSF (phenylmethyl sulfonyl fluoride)). After bringing the cells into contact with the lysis buffer, the solution is briefly sonicated, clarified by centrifugation, separated using SDS-PAGE (polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate), transferred to nitrocellulose or PVDF (polyvinylidene fluoride) and subjected to immunoblotting phospho-Akt (pAkt) S473, phospho-Akt T308, Akt and B-actin (Cell Signaling Technologies).

As a result, it was demonstrated that one or more of the compounds of the present description inhibit insulin stimulated Akt phosphorylation at position S473. Alternatively, certain compounds of this application further inhibit insulin stimulated Akt phosphorylation at position T308. This class of compounds can inhibit Akt more efficiently than rapamycin, and can be classified as mTORC2 inhibitors or upstream kinase inhibitors such as PI3K or Akt.

Example 32. Signaling in the blood through kinases

PI3K / Akt / mTor-mediated signal transmission is measured in blood cells using the phosflow method (Methods Enzymol. 2007, 434: 131-54). The advantage of this method is that, by its nature, it is a single cell analysis method, so it makes it possible to detect cellular heterogeneity, rather than population averages. This allows the simultaneous recognition of the state of signaling pathways in various populations identified by other markers. In addition, the phosflow method is a quantitative method with high sensitivity. To test the effect of one or more of the compounds provided herein, unfractionated splenocytes or peripheral blood mononuclear cells are stimulated with anti-CD3 antibodies to initiate signal transduction involving T cell receptors. After that, the cells are fixed and stained for surface markers and intracellular phosphoproteins. The inhibitors of this application are expected to inhibit the anti-CD3 mediated phosphorylation of Akt-S473 and S6, while rapamycin inhibits S6 phosphorylation and enhances Akt phosphorylation under test conditions.

Similarly, aliquots of whole blood are incubated for 15 minutes with a solvent (e.g. 0.1% DMSO) or kinase inhibitors at various concentrations, followed by the addition of stimulants to cross-crosslink the T cell receptor (TCR) (anti-CD3 with secondary antibody) or a B-cell receptor (BCR) using antibodies against the Kappa light chain (Fab ′ ₂ fragments). After about 5 and 15 minutes, the samples were fixed (for example, cold 4% paraformaldehyde) and used for the phosflow method. Surface staining is used to distinguish between T and B cells using antibodies to cell surface markers that are known in the art. The phosphorylation level of kinase substrates, such as Akt and S6, is then measured by incubating fixed cells with labeled antibodies specific for the phosphorylated isoforms of these proteins. The cell population is then analyzed using flow cytometry.

Example 33. Analysis of the formation of colonies

Mouse bone marrow cells immediately before that transformed with the p190-containing retrovirus BCR-Al (referred to in this application as p190-transduced cells) are seeded in the presence of various drug combinations for about 7 days on M3630 methylcellulose media with recombinant human IL-7 of approximately in 30% serum and count the number of colonies formed by visual observation with a microscope.

Alternatively, human peripheral blood mononuclear cells are obtained from patients with a positive Philadelphia chromosome (Ph +) and a negative Philadelphia chromosome (Ph-) according to an initial diagnosis or with a relapse of the disease. Living cells are isolated and enriched in CD19 + CD34 + B progenitor cells. After being left overnight in liquid culture, cells were plated on methocult GF + H4435 medium (Stem Cell Technologies) supplemented with cytokines (IL-3, IL-6, IL-7, G-CSF (granulocyte colony stimulating factor), GM-CSF (granulocyte -macrophage colony stimulating factor), CF, ligand for FIt3 and erythropoietin) and known chemotherapeutic agents in various concentrations in combination with any of the compounds of the present description. Colonies are counted after 12-14 days using a microscope. This method can be used to test evidence of cumulative or synergistic activity.

Example 34. The effect of kinase inhibitors on leukemia cells in vivo

Female recipient mice are irradiated with a lethal dose using a gamma radiation source in two separate doses with an interval of about 4 hours, about 5 Gy each. Approximately 1 hour after the second dose of radiation, mice are administered iv (intravenously) by injection of approximately 1 × 10 ⁶ leukemia cells (eg, human or mouse Ph + cells or p190-transduced bone marrow cells). These cells are administered together with an anti-radiation dose equal to about 5 × 10 ⁶ normal bone marrow cells from donor mice aged 3-5 weeks. Recipients are given antibiotics in water and monitored daily. Mice infected after approximately 14 days are euthanized and lymphoid organs removed for analysis. Treatment with kinase inhibitors begins about 10 days after the introduction of leukemia cells and continues daily until the mice become ill, or up to approximately 35 days after the introduction of the graft. Inhibitors are given using oral lavage.

Peripheral blood cells were harvested on approximately the 10th day (before treatment) and after euthanasia (after treatment), brought into contact with labeled anti-hCD4 antibodies (human CD4) and counted using flow cytometry. This method can be used to demonstrate that due to the synergistic effect of one or more of the compounds described in this application, in combination with known chemotherapeutic agents, the number of leukemia blood cells is significantly reduced compared to treatment with only known chemotherapeutic agents (e.g., Glivec) in testing conditions.

Example 35. Treatment of mice that are models of lupus disease

In mice lacking an inhibitory FcγRIIb receptor that inhibits PI3K-mediated signaling in B cells, lupus is highly likely to develop. FcγRIIb knockout mice (R2KO, Jackson Labs) are considered a suitable model for this disease in humans, as some patients with a diagnosis of lupus exhibit reduced expression or function of FcγRIIb (S. Bolland and J.V. Ravtech, 2000. Immunity 12: 277-285).

R2KO mice develop lupus-like disease with antinuclear antibodies, glomerulonephritis, and proteinuria at the age of about 4-6 months. For these experiments, the rapamycin analog RAD001 (supplied by LC Laboratories) was used as a reference compound and was administered orally. This compound has been shown to alleviate the symptoms of lupus in model B6.Sle1z.Sle3z (T. Wu et al. J. Clin. Invest. 117: 2186-2196).

Lupus model mice, such as R2KO, BXSB, or MLR / Ipr, are approximately 2 months old and are treated for approximately two months. Doses are given to mice: solvent, RAD001 (about 10 mg / kg) or the compounds described in this application (from about 1 mg / kg to about 500 mg / kg). For approximately the entire testing period, blood and urine samples are taken and analyzed for antinuclear antibodies (in dilutions in serum) or protein concentration (in urine). Serum is also tested for antibodies against ssDNA (single-stranded DNA) and against dsDNA (double-stranded DNA) by ELISA (enzyme-linked immunosorbent assay). Animals are euthanized on the 60th day and tissues are removed to determine the weight of the spleen and kidney disease. Glomerulonephritis is evaluated by staining sections of the kidneys with H&E (hematoxylin and eosin). Other animals are examined approximately two months after discontinuation of treatment using the same endpoints.

This model, recognized in the art, can be used to demonstrate that the kinase inhibitors set forth in this application can inhibit or delay the onset of lupus symptoms in mice that are models of lupus disease.

Example 36. Analysis of bone marrow transplant in mice

Female recipient mice are irradiated with a lethal dose using a γ-ray source. About 1 hour after the radiation dose, approximately 1 × 10 ⁶ leukemia cells from the early passages of p190 transduced cultures were injected into mice (for example, as described in Cancer Genet. Cytogenet. 2005 Aug, 161 (1): 51-6). These cells are administered together with an anti-radiation dose equal to about 5 × 10 ⁶ normal bone marrow cells from donor mice aged 3-5 weeks. Recipients are given antibiotics in water and monitored daily. Mice infected after approximately 14 days are euthanized and lymphoid organs are removed for flow cytometry and / or magnetic enrichment. Treatment begins on the 10th day and continues daily until the mice become ill, or up to approximately 35 days after transplant administration. Medicines are administered through a gastric tube (n / a). In a trial experiment, a dose of a chemotherapeutic agent is identified that is not therapeutic, but delays the onset of leukemia by about one week or less; as controls, treatment with a solvent or treatment with a chemotherapeutic agent is used, for which it was shown earlier in this model that it delays the onset of development but does not cure leukemogenesis (for example, imatinib at a dose of about 70 mg / kg twice a day). In the first step, p190-containing cells expressing eGFP (enhanced GFP) are used, and post-mortem analysis is limited to calculating the percentage of leukemia cells in the bone marrow, spleen, and lymph nodes (LN) using flow cytometry. In the second step, p190-bearing cells expressing the “tailless” form of human CD4 are used, and the post-mortem analysis involves magnetic sorting of hCD4 + cells from the spleen, followed by immunoblot analysis of key signaling endpoints: pAkt-T308 and -S473; pS6 and p4EVP-1. As controls for detection by immunoblotting, assorted cells are incubated before lysis in the presence or absence of the kinase inhibitors of the present description. If desired, the phosflow method is used to detect pAkt-S473 and pS6-S235 / 236 in hCD4-gated cells without prior sorting. Such signal transmission studies are especially useful if, for example, mice receiving drug therapy do not develop clinical leukemia at a point in time of 35 days. Kaplan-Meyer survival curves are plotted and statistical analysis is performed using methods known in the art. Results for p190-containing cells are analyzed individually as well as cumulatively.

Once a week, peripheral blood samples (100-200 μl) were taken from all mice, starting from 10 days immediately before treatment. Blood plasma is used to measure drug concentrations and cells are analyzed for leukemia markers (eGFP or hCD4) and signal transmission biomarkers as set forth in this application.

This general analysis, known in the art, can be used to demonstrate that effective therapeutic doses of the compounds of this application can be used to inhibit the proliferation of leukemia cells.

Example 37. Analysis of T-cell independent B cell activation by TNP-ficoll (TNP-F)

To test the effect of the compounds of the present invention on the suppression of T-cell independent antibody production, B-cell activation analysis by TNP-ficoll was used as set forth in this application. The compounds of the present invention were dissolved in an appropriate solvent (for example, containing 5% 1-methyl-2-pyrrolidinone, 85% polyethylene glycol 400, 10% solutor). Compounds were administered orally to mice 4-10 weeks old approximately 1 hour before treatment with TNP-ficoll. To study the effect of these compounds on B-cell activation, one batch of mice was grouped according to the following table:

Group number No. of mice / group Compound administered for treatment Group Antigen Injection on Day 1 The introduction of the compound, starting from 1 day to 7 days TNP-F Way (mg / kg) Way Mode one four Solvent Antigen only 0 2 8 - Antigen only 0 2 times a day for 7 days 3 8 Compound No. 7 Reference 200 μl (0.5 mg / ml) in / b thirty by four 8 one 5 8 Compound No. 53 Antigen + Comp. 3 6 8 10 7 8 thirty 8 8 60

Four animals in group 1 and eight animals in groups 2-7 were euthanized in CO ₂ 2 hours after the last administration of the compounds on the 7th day. Immediate blood sampling was performed by heart puncture and the blood was stored at 37 ° C for 1 h so that it coagulated, after which it was incubated overnight at 4 ° C to reduce the thrombus. The next day, serum was collected by decantation and centrifugation at 3000 rpm for 10 minutes. The collected serum was then frozen at -80 ° C for further analysis.

Serum samples were analyzed by ELISA for anti-TNP antibody titers as set forth herein. Wells of a Nunc Maxisorb microtiter plate were coated with 100 μl / well TNP-BSA at a concentration of 10 μg / ml in phosphate buffered saline (PBS). The Maxisorb plate was incubated for 1.5 hours at room temperature and the solution was removed. 200 μl / well of blocking buffer (for example, 1% BSA in PBS) was added to each well and incubated for 1 h at room temperature. The plate was washed once with 200 μl / well of 0.05% Tween-20 in PBS (wash buffer). Diluted 1: 2 serum in blocking buffer from each mouse was added to each well of the first column (1) of the microtiter plate. The serum in each well of column 1 was then diluted 3 times in blocking buffer and added to column 2. The serum in each well of column 2 was diluted 3 times in blocking buffer and added to column 3. This procedure was repeated in all twelve columns of the microtiter plate. The microtiter plate was incubated for 1 h at room temperature. The serum was removed from the plate and the plate was washed three times with wash buffer. 100 μl / well HRP (horseradish peroxidase) conjugated goat anti-mouse IgG3 antibodies diluted 1: 250 in blocking buffer were added to each well and incubated for 1 h at room temperature. HRP-conjugated anti-IgG3 mouse antibodies were removed from the microtiter plate and the plate was washed six times with wash buffer. HRP substrate (200 μl ABTS (2,2'-azino-bis- (3-zylbenzothiazoline-6-sulfonic acid) + 30% H ₂ O ₂ + 10 ml citrate buffer) was added to each well in 100 μl / well were incubated for 2-20 minutes in the dark and the amount of anti-TNP IgG3 was determined spectrophotometrically at 405 nm Similarly, the amount of anti-TNP IgM and the total amount of anti-TNP Al (antibodies) were determined using HRP conjugated anti-mouse IgM antibodies and conjugated with HRP antibodies against mouse Ig, respectively.

The results shown in Figure 2 also show that under testing conditions, compounds No. 7 and No. 53 show a 3.4- and 6.5-fold decrease, respectively, in IgG3 levels in mice compared to the solvent as a control, at a level doses of 30 mg / kg. Figure 2 also shows that under testing conditions, compound No. 53 shows a 29.9-fold decrease in IgG3 levels in mice compared to the solvent as a control, at a dose level of 60 mg / kg.

Example 38. Analysis of developing collagen-induced type II arthritis in rats

In order to study the effect of the compounds of the present invention on autoimmune arthritis disease, a model of developing collagen-induced arthritis was used. Female Lewis rats were injected with collagen on day 0. Bovine type II collagen was prepared as a solution at a concentration of 4 mg / ml in 0.01 N. acetic acid. Equal volumes of a solution of collagen and Freud's incomplete adjuvant were emulsified, mixing by hand until a drop of emulsifiable substance began to retain its shape in water. Each rodent was injected with a subcutaneous injection of 300 μl of the mixture, at each time the injection was administered, distributing it in three areas on the back.

Oral administration of the compounds began on day 0 and continued on day 16 inclusive, introducing a solvent (5% NMP, 85% PEG 400, 10% Solutol) or the compounds of the present invention in a solvent or control (e.g. methotrexate) from 12 at hourly intervals once a day. Rats were weighed on days 0, 3, 6, 9-17, and ankle joints were measured using a caliper on days 9-17. The final measurement of body weight was carried out and then the animals were euthanized on the 17th day. After euthanasia, blood was taken and the hind legs and knee joints were separated. Next, the blood was processed for pharmacokinetic experiments, as well as ELISA analysis for antibodies against type II collagen. The hind legs were weighed and then, together with the knee joints, were stored in 10% formalin. The paws and knee joints were further processed for microscopy. The liver, spleen, and thymus were also weighed. The sciatic nerves were prepared for histopathological examination.

The knee and ankle joints were fixed for 1-2 days and decalcified for 4-5 days. Ankle joints were cut into two equal parts in the longitudinal direction, knee joints were cut into two equal parts along the frontal plane. Then the joints were processed, fixed, sections were made and stained with toluidine blue. Joints were evaluated in accordance with the following criteria.

Inflammation of the knee and ankle joints

0 - normal.

1 - minimal infiltration of inflammatory cells into the synovial / periarticular tissue.

2 - easy infiltration.

3 - moderate infiltration with moderate edema.

4 - marked infiltration with marked swelling.

5 - severe infiltration with severe edema.

Ankle Pannus

0 - normal.

1 - minimal infiltration of the pannus into the cartilage and the cartilage bone.

2 - easy infiltration (less than 1/4 of the tibia or tarsus in the marginal zones).

3 - moderate infiltration (1 / 4-1 / 3 of the tibia or small bones of the tarsus in the marginal zones).

4 - marked infiltration (1 / 2-3 / 4 of the tibia or tarsus in the marginal zones).

5 - severe infiltration (more than 3/4 of the tibia or tarsus in the marginal zones, severe violation of the general structure).

Pannus of the knee joint

0 - normal.

1 - minimal infiltration of the pannus into the cartilage and the cartilage bone.

2 - mild infiltration (extends up to 1/4 of the surface or the cartilage of the tibia or femur).

3 - moderate infiltration (spreads in the range from more than 1/4, but less than 1/2 of the surface or the cartilage of the tibia or femur).

4 - marked infiltration (spreads in the range from 1/2 to 3/4 of the surface of the tibia or femur).

5 - severe infiltration (covers more than 3/4 of the surface).

Cartilage affection (ankle joint, with emphasis on small

tarsal bones)

0 - normal.

1 - minimal - weakening of the color with toluidine blue from minimal to mild without any visible decrease in the number of chondrocytes or collagen destruction.

2 - mild - mild degree of weakening of the color with toluidine blue with a light (superficial), focal character, a decrease in the number of chondrocytes and / or collagen destruction.

3 - moderate - moderate degree of weakening of the color with toluidine blue with moderate (to a thickness to the middle of the zone), multi-focal nature, a decrease in the number of chondrocytes and / or collagen destruction, with damage to small bones of the tarsus to 1 / 2-3 / 4 depths.

4 - noticeable - noticeable weakening of the color with toluidine blue with a noticeable (to a thickness to a deep zone), multi-focal nature, a decrease in the number of chondrocytes and / or collagen destruction, with the loss of cartilage throughout the thickness of 1 or more small bones of the tarsus.

5 - severe - significant diffuse weakening of the color with toluidine blue with a significant (by thickness to the demarcation line), multi-focal nature, a decrease in the number of chondrocytes and / or collagen destruction.

Cartilage affection (knee joint, with emphasis on the condyles of the femur).

0 - normal.

1 - minimal - weakening of the color with toluidine blue from minimal to mild without any visible decrease in the number of chondrocytes or collagen destruction.

2 - mild - mild degree of weakening of the color with toluidine blue with a light (superficial), focal character, a decrease in the number of chondrocytes and / or collagen destruction.

3 - moderate - moderate degree of weakening of the color with toluidine blue with moderate (by thickness to the middle of the zone), from multi-focal to diffuse, with a decrease in the number of chondrocytes and / or collagen destruction.

4 - noticeable - noticeable weakening of the color with toluidine blue with a noticeable (by thickness to a deep zone), from multi-focal to diffuse nature, a decrease in the number of chondrocytes and / or destruction of collagen on the surface of one femur with a complete loss or loss close to full.

5 - severe - significant diffuse weakening of the color with toluidine blue with a significant (by thickness to the demarcation line), multi-focal nature, a decrease in the number of chondrocytes and / or collagen destruction on both thigh bones and / or tibia.

Bone resorption (ankle joint)

0 - normal.

1 - minimal - small areas of resorption, hardly distinguishable at low magnification, rare osteoclasts.

2 - easy - more numerous areas of resorption, hardly distinguishable at low magnification, osteoclasts are more numerous, less than 1/4 of the tibia or tarsal bones in the marginal zones are resorbed.

3 - moderate - clearly noticeable resorption of the medullary trabecular or cortical bone without deep, to the entire thickness, defects of the surface layer, loss of some medullary trabeculae, the lesion is distinguishable at low magnification, osteoclasts are more numerous, from 1/4 to 1/3 of the tibia or bones of the tarsus in the marginal zones.

4 - noticeable - deep, over the entire thickness, defects of the cortical bone, often with deformation of the profile of the rest of the cortical surface, a noticeable loss of the medullary part of the bone, numerous osteoclasts, affected 1 / 2-3 / 4 of the tibia or tarsal bones in the marginal zones.

5 - strong - deep, over the entire thickness, defects of the cortical bone, often with deformation of the profile of the rest of the cortical surface, a noticeable loss of the medullary part of the bone, numerous osteoclasts, more than 3/4 of the tibia or tarsal bones in the marginal zones, severe deformation of the general structure .

Bone resorption (knee joint)

0 - normal.

1 - minimal - small areas of resorption, hardly distinguishable at low magnification, rare osteoclasts.

2 - mild - more numerous areas of resorption, some loss of the subchondral bone, including 1/4 of the surface of the tibia or femur (medial or lateral).

3 - moderate - clearly noticeable resorption of the cartilage, including more than 1/4, but less than 1/2 of the surface of the tibia or femur (medial or lateral).

4 - noticeable - clearly noticeable resorption of the cartilage, including at least 1/2, but less than 3/4 of the surface of the tibia or femur (medial or lateral).

5 - severe - deformation of the entire joint due to destruction involving more than 3/4 of the surface of the tibia or femur (medial or lateral).

Statistical analysis of body weight / paws, AUC parameters (area under the curve) for paws and histopathological parameters were evaluated using Student's t-test or other relevant criteria (ANOVA ANOVA analysis with post-experimental test), with a significance level of 5%. The percent inhibition of paw mass and AUC was calculated using the following formula:

% inhibition = (AB / A) X100, where

And represents the average value for the control objects of the disease minus the average value for the norm;

B represents the average value for the treated objects minus the average value for the norm.

The results shown in FIG. 3 demonstrate the effect of compound No. 53 at doses of 10, 30 and 60 mg / kg at 12-hour intervals under conditions of testing on the average diameter of the ankle joint in time in a model of developing collagen-induced type II arthritis in rats. Compared to the control in the form of a solvent alone or the control in the form of methotrexate, the compounds of the present invention showed a significant decrease in arthritis-induced increase in the diameter of the ankle joint over time.

The results shown in Figure 4 demonstrate the effect of compounds No. 7 and No. 53 on histopathology of the ankle joint in the categories: inflammation, pannus, cartilage damage and bone resorption, as described previously in the test conditions. The results indicate a significant decrease in one or more categories under the influence of one of the compounds of the present invention (i.e., compound No. 53) under test conditions. Figure 4 also shows that at a dose of 60 mg / kg under test conditions, there is a statistically significant decrease in all categories of histopathology of the ankle joint detected for one of the compounds of the present invention (i.e., compound No. 53). This is evidence that one or more of the compounds of the present invention may be useful in treating and alleviating symptoms of arthritis.

The results shown in FIG. 5 demonstrate the effect of compounds No. 7 and No. 53 under testing conditions on histopathology of the knee joint. The results demonstrate a dose-dependent decrease in knee histopathology. This is evidence that one or more of the compounds of the present invention may be useful in treating and alleviating symptoms of arthritis.

The results shown in FIG. 6 demonstrate the effect of compounds No. 7 and No. 53 under test conditions on serum levels of antibodies against type II collagen. The results also indicate a significant decrease in serum antibodies against type II collagen in blood serum at dose levels of compound No. 53 equal to 10, 20 and 60 mg / kg, which confirms that one or more compounds of the present invention can be useful not only to treat and alleviate the symptoms of arthritis, but may also be useful in inhibiting the autoimmune reaction itself.

The results shown in Fig.7 demonstrate the effect of compound No. 7 at doses of 10, 30 and 60 mg / kg at 12-hour intervals under conditions of testing on the average diameter of the ankle joint in time. Compared to the control in the form of a solvent alone or the control in the form of methotrexate, the compound under test conditions showed a significant decrease in arthritis-induced increase in the diameter of the ankle joint over time.

Example 39. Analysis of established collagen-induced type II arthritis in rats

In order to study the dose-dependent efficacy of the compounds of the present invention in inhibiting inflammation, cartilage destruction and bone resorption with arthritis type II-induced collagen established on day 7, the compounds were orally administered once or twice a day for 6 days in rats.

Female Lewis rats were anesthetized and injected with collagen prepared and administered as described previously on day 0. On the 6th day, the animals were anesthetized and a second injection of collagen was made. Measurements with a caliper of normal (before the disease) of the right and left ankle joints were performed on the 9th day. Typically, arthritis appeared on days 10-11, and rats were randomized to treatment groups. Randomization was performed after an ankle swelling was clearly established and had good evidence of a bilateral disease.

After selection of animals for participation in this study, treatment was initiated using the oral route of administration. Animals received a solvent, control (enbrel), or doses of the compounds twice a day or once a day (2x / day or 1x / day, respectively). The introduction was carried out on days 1-6, using a volume of 2.5 ml / kg (2x / day) or 5 ml / kg (1x / day) for oral solutions. Rats were weighed on days 1-7 after the establishment of arthritis and every day ankle joints were measured using a caliper. The final values of body weight were determined on the 7th day and the animals were euthanized.

The results shown in Fig. 8 show a significant decrease in the increase in the average diameter of the ankle joint in time for compound No. 53 when it is administered once a day under testing conditions. The results shown in FIG. 9 also show a significant decrease in the increase in the average diameter of the ankle joint over time for compound No. 53 when administered twice a day under test conditions. This confirms that the compounds of the present invention may be useful in the treatment of autoimmune diseases such as arthritis.

Example 40. Analysis of Arthritis Induced by Adjuvant

Rat Intrathecal Catheterization

An anesthetized isoflurane rat Lewis rats (200-250 g) were implanted intrathecal (and / t) catheter. After a recovery period of 6 days, all animals were used for experiments, with the exception of those that showed anomalies in sensitivity or motor activity (less than 5% of the total). To carry out and / t administration, 10 μl of the drug or physiological saline was injected through the catheter, then 10 μl of isotonic saline.

Adjuvant Induced Arthritis and Drug Therapy

Lewis rats were immunized at the base of the tail using 0.1 ml of complete Freund's adjuvant (CFA), on day 0 several days after catheter implantation (n is 6 / group). Drug therapy (for example, with one or more than one compound of the present invention or with a solvent) was usually started on the 8th day and continued daily up to 20 days. Clinical signs of arthritis usually appear on the 10th day, and paw swelling was determined every second day using water displacement plethysmometry.

The results shown in Fig. 10, by means of an average change in the paw volume for the indicated administration schemes, show that under test conditions, compound No. 53 shows a dose-dependent decrease in the increase in the average paw volume according to the measurement results in this model system of adjuvant-induced arthritis. These results confirm that one or more of the compounds of the present invention may be useful in treating one or more of the diseases or conditions described herein.

The results shown in Fig. 11 show that compound No. 53 does not exhibit toxicity or other adverse reactions under test conditions, which is estimated by the absence of weight reduction.

Example 41. Pharmacokinetic analysis in rodents

In order to study the pharmacokinetics of the compounds of the present invention from a batch of mice aged 4-10 weeks, the groups are composed in accordance with the following table:

Group No. The number of mice / group Compound administered for treatment Administration of the compound starting from day 1 to day 7 (mg / kg) Way Mode one 3 one by 2 times a day for 7 days 2 3 3 3 3 10 four 3 thirty 5 3 60

The compounds of the present invention are dissolved in an appropriate carrier (for example, containing 5% 1-methyl-2-pyrrolidinone, 85% polyethylene glycol 400, 10% Solutor) and administered orally at intervals of 12 hours once a day. All animals were euthanized in CO ₂ 2 hours after the final administration of the compound. Immediate blood sampling is performed and stored on ice to isolate plasma. Plasma is isolated by centrifugation at 5,000 rpm for 10 minutes. The collected plasma is frozen for pharmacokinetic determination.

The results are expected to demonstrate pharmacokinetic parameters, such as absorption, distribution, metabolism, excretion and toxicity parameters for the compounds of the present invention.

Example 42. Bazotest (Vasotest) analysis (analysis using the test of activation of basophils by allergens)

The Basotest assay is performed using Orpegen Pharma's Basotest reagent kit. Whole heparinized blood is pre-incubated with the test compound or solvent at 37 ° C for 20 minutes. Next, the blood is incubated with a stimulating buffer from the assay kit (to prime cells for an answer), then with an allergen (dust extract containing dust mite, or plant extract) for 20 minutes. The degranulation process is stopped by incubating blood samples on ice. After that, a label is introduced into the cells using anti-IgE-PE (phycoerythrin) to detect basophilic granulocytes, and anti-gp53-FITC to detect gp53 (glycoprotein expressed on activated basophils). After staining, red blood cells are lysed by adding a lysis solution. Cells were washed and analyzed using flow cytometry. Compounds 7 and 53, when tested in this assay, inhibit allergen-induced activation of basophilic granulocytes in the range of submicromolar concentrations.

Example 43. Combined Use of PI3Kδ Inhibitors and Agents That Inhibit IgE Production or Activity

The compounds of the present invention may exhibit synergistic or additive efficacy when administered in combination with agents that inhibit the production or activity of IgE.

Agents that inhibit the production of IgE include, for example, one or more of TEI-9874, 2- (4- (6-cyclohexyloxy-2-naphthyloxy) phenylacetamide) benzoic acid, rapamycin, rapamycin analogs (i.e., rapalogs), TORC1 inhibitors, TORC2 inhibitors, and any other compounds that inhibit mTORC1 and mTORC2. Agents that inhibit IgE activity include, for example, anti-IgE antibodies, such as, for example, omalizumab and TNX-901.

One or more compounds of the invention capable of inhibiting PI3Kδ are effective in the treatment of autoimmune and inflammatory disorders (AMD), for example, rheumatoid arthritis. If any of the compounds causes an undesired level of IgE production, it can be selected for administration in combination with an agent that inhibits IgE production or IgE activity. In addition, the administration of the PI3K5 or PI3Kb / y inhibitors of the present invention in combination with mTOR inhibitors can also lead to synergism through enhanced inhibition of the PI3K pathway. Various in vivo and in vitro models can be used to establish the effect of such combination treatment on AMD, including, but not limited to, (a) analysis of antibody production in vitro by B cells, (b) in vivo TNP analysis, and (c) collagen-induced arthritis model in rodents.

(a) Analysis using b-cells

Mice are euthanized, spleens are removed and ground through a nylon mesh to obtain a suspension of individual cells. Splenocytes are washed (after removal of red blood cells by osmotic shock) and incubated with microbeads conjugated with anti-CD43 and anti-Mac-1 antibodies (Miltenyi Biotec). Cells bound to the granules are separated from unbound cells using a magnetic cell sorter. Unnecessary cells are held in a magnetized column, and the remaining B cells are collected in a flowing stream. The purified B cells are stimulated with a lipopolysaccharide or anti-CD40 antibody and interleukin 4. The stimulated B cells are treated only with a solvent or solvent together with the PI3Kδ inhibitors of the present invention, such as compound 53, with mTOR inhibitors such as rapamycin, rapalogi or mTORC1 / inhibitors C2, or without them. Based on these results, it is expected that, in the presence of only mTOR inhibitors (e.g. rapamycin), the IgG and IgE response will be in the range from small to substantially zero. However, in the presence of PI3Kδ and mTOR inhibitors, B cells will exhibit a weakened IgG response compared to solvent-treated B cells, and B cells are expected to exhibit a weakened IgE response compared to the response from B cells treated only with PI3Kδ inhibitors.

(b) TNP analysis

Mice were immunized with TNP-ficoll or TNP-KHL (fisurelia mollusk hemocyanin) and were treated with: a solvent, a PI3K6 inhibitor, for example, compound 53 of the present invention, an mTOR inhibitor, for example rapamycin, or a PI3Kδ inhibitor in combination with an mTOR inhibitor such as rapamycin. Serum levels of antigen-specific IgE are measured by ELISA using TNP-BSA coated plates and isotype-specific labeled antibodies. Mice treated only with the mTOR inhibitor are expected to show little or no effect on the antigen-specific IgG3 response and no statistically significant increase in the IgE response compared to the solvent as a control. It is also expected that mice treated with PI3Kδ inhibitors and mTOR inhibitors will show a decrease in antigen-specific IgG3 response compared to solvent-treated mice. In addition, mice treated with both PI3Kδ inhibitors and mTOR inhibitors will show an increase in IgE response compared to mice treated with PI3Kδ inhibitor only.

(c) Collagen-Induced Arthritis Model in Rats

Female Lewis rats are anesthetized and collagen injections are made on day 0, which are prepared and administered as described previously. On the 6th day, the animals are anesthetized and a second injection of collagen is given. Measurements with a caliper of normal (before the disease) of the right and left ankle joints are carried out on the 9th day. Typically, arthritis appears on days 10-11 and rats are randomized to treatment groups. Randomization is carried out after an ankle swelling is clearly established and there is good evidence of a bilateral disease.

After selecting animals to participate in this study, treatment is started. Animals receive a solvent, PI3Kδ inhibitor, or PI3Kδ inhibitor in combination with rapamycin. The introduction is carried out on the 1-6th day. Rats are weighed on days 1-7 after the establishment of arthritis and every day ankle joints are measured using a caliper. The final values of body weight are determined on the 7th day and the animals are euthanized.

Combined treatment with a PI3Kδ inhibitor and rapamycin is expected to be more effective than treatment with a PI3Kδ inhibitor alone.

Although preferred embodiments of the present invention are shown and set forth herein, it will be apparent to those skilled in the art that such embodiments are given by way of example only. Specialists in the art will immediately have the ideas of numerous variations, changes and substitutions without deviating from the invention. It should be understood that in the practical application of the invention, various alternatives to the embodiments of the invention set forth in this application may be used. It is intended that the appended claims define the scope of the invention and thereby cover methods and structures falling within the scope of this claims and their equivalents.

Claims (24)

1. The compound of formula I:

or its pharmaceutically acceptable salt, where
B is a grouping of formula II:

where W _c represents a 6-10 membered aryl; and
q is an integer of 0, 1, 2, 3 or 4;
X is absent or represents - (CH (R ⁹ )) _z -, and z is an integer of 1;
Y represents —N (R ⁹ ) -;
W _d represents

R ¹ represents C _1-6 alkyl or halogen;
R ² represents C _1-6 alkyl or halogen;
R ³ represents hydrogen or C _1-6 alkyl; and
in each case, R ⁹ independently represents hydrogen or C _1-6 alkyl. 2. The compound according to claim 1, where q is 0. 3. The compound according to claim 2, where R ¹ represents halogen. 4. The compound according to claim 2, where R ¹ represents C _1-6 alkyl. 5. The compound according to claim 4, where R ¹ represents methyl. 6. The compound according to claim 1, where R ³ represents C _1-6 alkyl. 7. The compound according to claim 6, where R ³ represents methyl. 8. The compound according to claim 1, where R ⁹ represents hydrogen. 9. The compound according to claim 1, where R ⁹ represents methyl. 10. The compound according to claim 1, where R ⁹ represents ethyl. 11. The compound according to claim 1 or its pharmaceutically acceptable salt, where the compound is a

12. A pharmaceutical composition having phosphatidylinositol-3-kinase (PI3 kinase) inhibitory activity, comprising a pharmaceutically acceptable excipient and a therapeutically effective amount of a compound according to any one of claims 1 to 11. 13. The composition according to p. 12, in liquid, solid, semi-solid form, gel form or aerosol form. 14. A method of inhibiting PI3 kinase in a subject, comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 11. 15. The method according to 14, where the subject is suffering from a disease, which is a cancer, bone disorder, inflammatory disease, immune disease, nervous system disease, metabolic disease, respiratory disease, thrombosis or cardiovascular disease. 16. The method according to clause 15, where the subject is suffering from cancer. 17. The method according to clause 16, where the cancer is breast cancer, ovarian cancer, uterine cancer, cervical cancer, prostate cancer, pancreatic cancer, bladder cancer, bone cancer, lung cancer, skin cancer, ocular retinoblastoma, primary liver cancer, kidney cancer, thyroid cancer, virus-induced cancer, central nervous system cancer, peripheral nervous system cancer, oral and oropharyngeal cancer, stomach cancer, testicular cancer, thymus cancer, colon cancer or colon cancer. 18. The method of claim 17, wherein the lung cancer is non-small cell lung cancer (NSCLC) or small cell lung cancer. 19. The method according to clause 16, where the cancer is lymphoma or leukemia. 20. The method according to claim 19, where the lymphoma is a diffuse b-large cell lymphoma, b-cell immunoblastic lymphoma and small cell lymphoma from undigested cells. 21. The method according to claim 19, where the leukemia is acute myeloid leukemia (AML), acute lymphocytic leukemia, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia, hairy cell leukemia, myelodysplasia, myeloproliferative disorder, mastocytosis, multiple myeloma ( myelodysplastic syndrome (MDS). 22. The method according to clause 16, where the cancer is cutaneous or intraocular melanoma (eye melanoma), Kaposi’s sarcoma, adult T-cell leukemia or adult T-cell lymphoma. 23. The method of claim 14, wherein a second therapeutic agent is administered in combination with the drug. 24. The method according to item 23, where the second agent is an agent that inhibits the production of IgE.

Images