AKT3 Modulator and How to Use It

Abstract

This invention provides methods for treating and preventing various diseases using Akt3 activators and inhibitors. [Solution] A method for treating a disease in a subject requiring treatment, comprising administering to the subject a composition containing, for example, an Akt3 modulator represented by the following formula IV, in an amount effective to modulate Akt3 signaling and treat or delay the progression of the disease. JPEG2026143396000030.jpg4668

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims the benefit and priority of U.S. Provisional Application 63 / 021,797 filed 8 May 2020, and incorporates the contents of this application in whole by reference.

[0002] Inclusion by citation Any patents, patent publications, publications, or other documents cited herein are expressly incorporated herein by their entirety.

[0003] Field of Invention The present invention relates to a method for treating and preventing diseases by generally regulating Akt3 signaling. [Background technology]

[0004] Background of the Invention Chronic diseases and illnesses are persistent conditions that require ongoing treatment and generally negatively impact a patient's quality of life. Chronic diseases are the leading causes of disability and death in the United States. Common chronic diseases include, but are not limited to, inflammatory diseases, neurodegenerative diseases, pathogenic infections, immunodeficiency disorders, weight loss disorders, hormonal imbalances, tuberous sclerosis, retinitis pigmentosa, and congestive heart failure. It is estimated that roughly 6 out of 10 adults in the United States have a chronic disease, and 4 out of 10 have two or more chronic diseases. Chronic diseases are also a leading driver of $3.3 trillion in annual healthcare spending in the United States (see National Center for Chronic Disease Prevention and Health Promotion). These astonishing statistics highlight the need for new and improved treatments and preventive interventions for chronic diseases and illnesses.

[0005] Neurodegenerative diseases are incurable, debilitating conditions characterized by the progressive degeneration and cell death of nerve cells, also known as neurons. Neurons are components of the nervous system and, normally, do not regenerate or replace themselves when damaged or dead. Neuronal loss or dysfunction in patients with neurodegenerative diseases can affect body movement and brain function. Common neurodegenerative diseases include, but are not limited to, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, Parkinson's disease, multiple sclerosis, prion diseases, motor neuron disorders, spinocerebellar ataxia, and spinal muscular atrophy. Symptoms of advanced neurodegenerative diseases can be devastating, causing patients to lose memory, motor control, and personality. Currently, neurodegenerative diseases are incurable, and treatments focused on managing symptoms typically result in negative side effects that impair the patient's quality of life.

[0006] A serious complication of chronic diseases such as neurodegenerative diseases is cachexia or wasting syndrome. Cachexia is defined as a weight loss of more than 5% of body weight within 12 months in the presence of a chronic disease. Other symptoms of cachexia include muscle atrophy, fatigue, weakness, and often loss of appetite. The weight loss associated with cachexia is due to a decrease in muscle mass as well as fat. Patients with cachexia often lose weight even if they are still eating a normal diet. There is currently no effective treatment for cachexia, which leads to many chronic disease-related deaths.

[0007] The need for more effective and tolerable treatments and preventive interventions for chronic diseases and their associated complications continues to grow. [Overview of the initiative] [Problems that the invention aims to solve]

[0008] Therefore, the object of the present invention is to provide methods for treating and preventing chronic diseases.

[0009] Another objective of the present invention is to provide methods for treating and preventing complications of chronic diseases such as cachexia. [Means for solving the problem]

[0010] Summary of the Invention Compounds and pharmaceutical compositions that selectively modulate Akt3 are disclosed herein. Methods of using the compounds to treat or prevent various diseases and disorders are also disclosed. Non-limiting examples of diseases include inflammatory diseases, neurodegenerative diseases, pathogenic infections, immunodeficiency disorders, weight loss disorders, hormonal imbalances, tuberous sclerosis, retinitis pigmentosa, congestive heart failure, and combinations thereof.

[0011] Akt3 is highly expressed in the brain, and its dysregulation is associated with several neurodegenerative diseases. Therefore, the disclosed Akt3 modulator may be useful in treating neurodegenerative diseases.

[0012] Methods for treating neurodegenerative diseases in subjects requiring treatment are disclosed. In one embodiment, the method involves administering to a subject an Akt3 activator in a dose effective for neuroprotective signaling downstream of Akt3, thereby activating Akt3 in brain tissue. In another embodiment, the method involves administering to a subject an Akt3 inhibitor in a dose effective for neuroinflammatory signaling downstream of Akt3, thereby inhibiting Akt3 in brain tissue.

[0013] In one embodiment, the neurodegenerative disease to be treated is an acute neurodegenerative disease selected from the group consisting of epilepsy, transient ischemia of the spinal cord, or cerebral ischemia. The neurodegenerative disease may also be a chronic neurodegenerative disease selected from the group consisting of Huntington's disease, Alzheimer's disease, Parkinson's disease, multiple sclerosis, spinal muscular atrophy, or amyotrophic lateral sclerosis.

[0014] Akt3 is also highly expressed in adipose tissue and adipocytes. Akt3 signaling is associated with adipogenesis. Therefore, the disclosed Akt3 modulator may be useful in treating diseases and disorders characterized by excessive weight loss.

[0015] A method for treating excessive weight loss and eating disorders associated with conditions such as cachexia is disclosed. The method involves targeting an Akt3 inhibitor at a dose effective in inhibiting Akt3 signaling in adipocytes and promoting adipogenesis.

[0016] In one embodiment, the subject has excessive weight loss due to cachexy. Cachexy may be associated with chronic diseases such as acquired immunodeficiency syndrome (AIDS), celiac disease, chronic obstructive pulmonary disease, multiple sclerosis, rheumatoid arthritis, congestive heart failure, tuberculosis, familial amyloid polyneuropathy, Crohn's disease, untreated and severe type 1 diabetes mellitus, anorexia nervosa, hyperthyroidism, and hormone deficiencies.

[0017] The disclosed methods may include administering a second treatment agent to the subject. For neurodegenerative diseases, the second treatment agent may be an antispasmodic, muscle relaxant, analgesic, antidepressant, antipsychotic, anticonvulsant, anticholinergic, or anxiolytic. For subjects with excessive weight loss, the second treatment agent may be an appetite stimulant, nutritional supplement, 5-HT3 antagonist, or Cox-2 inhibitor.

[0018] In one embodiment, a method for treating a disease in a subject requiring treatment is described, comprising administering to the subject a composition comprising an Akt3 modulator in an amount effective to modulate Akt3 signaling and treat or delay the progression of the disease.

[0019] In any of the embodiments described herein, the disease is selected from the group consisting of neurodegenerative diseases, cachexia, eating disorders, obesity complications, inflammatory diseases, virus-induced inflammatory responses, Gulf War syndrome, tuberous sclerosis, retinitis pigmentosa, graft rejection, cancer, ischemic tissue injury, traumatic tissue injury, and combinations thereof.

[0020] In any of the embodiments described herein, the disease is a neurodegenerative disease.

[0021] In any of the embodiments described herein, the neurodegenerative disease is selected from the group consisting of Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, motor neuron disorders, Huntington's disease, HIV-induced neurodegeneration, Lewy body disease, spinal muscular atrophy, prion disease, spinocerebellar ataxia, familial amyloid polyneuropathy, and combinations thereof.

[0022] In any of the embodiments described herein, the disorder is cachexia or an eating disorder.

[0023] In any of the embodiments described herein, the disease is an obesity complication.

[0024] In any of the embodiments described herein, obesity complications are selected from the group consisting of glucose intolerance, fatty liver, dyslipidemia, and combinations thereof.

[0025] In any of the embodiments described herein, the disease is an inflammatory disease.

[0026] In any of the embodiments described herein, the inflammatory disease is selected from the group consisting of atopic dermatitis, allergies, asthma, and combinations thereof.

[0027] In any of the embodiments described herein, the disease is a virus-induced inflammatory response.

[0028] In any of the embodiments described herein, the virus-induced inflammatory response is SARS-induced inflammatory pneumonia, coronavirus disease 2019, or a combination thereof.

[0029] In any of the embodiments described herein, the disease is Gulf War Syndrome or tuberous sclerosis.

[0030] In any of the embodiments described herein, the disease is retinitis pigmentosa or graft rejection.

[0031] In any of the embodiments described herein, the disease is ischemic tissue injury or traumatic tissue injury.

[0032] In any of the embodiments described herein, the disease is cancer.

[0033] In any of the embodiments described herein, cancer is selected from the group consisting of adult T-cell leukemia / lymphoma, bladder, brain, breast, cervix, colorectal, esophagus, kidney, liver, lung, nasopharynx, pancreas, prostate, skin, stomach, uterus, ovaries, and testes.

[0034] In any of the embodiments described herein, cancer is leukemia.

[0035] In any of the embodiments described herein, leukemia is adult T-cell leukemia / lymphoma.

[0036] In any of the embodiments described herein, adult T-cell leukemia / lymphoma is caused by a human T-cell lymphotropic virus.

[0037] In any of the embodiments described herein, Akt3 is regulated by immune cells.

[0038] In any of the embodiments described herein, the immune cells are selected from the group consisting of T cells, B cells, macrophages, and glial cells.

[0039] In any of the embodiments described herein, the glial cell is an astrocyte, microglia, or oligodendrocyte.

[0040] In any of the embodiments described herein, the T cell is a T regulatory cell.

[0041] In any of the embodiments described herein, the Akt3 modulator activates Akt3 signaling.

[0042] In any of the embodiments described herein, the Akt3 modulator inhibits Akt3 signal transduction.

[0043] In any of the embodiments described herein, the Akt3 modulator increases T regulatory cell activity or production.

[0044] In any of the embodiments described herein, the Akt3 modulator decreases T regulatory cell activity or production.

[0045] In any of the embodiments described herein, the modulator of Akt3 is a compound of Formula I:

化

[0046] In any of the embodiments described herein, the Akt3 modulator is Equation II: [ka] a compound of or a pharmaceutically acceptable enantiomer, salt or solvate thereof, wherein: R1 is optionally -(C1-C 12 )-alkyl, -(C3-C 12 )-cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -O-(C1-C 12 )-alkyl, -O-(C1-C 12 )-alkyl-(C6-C 20 )-aryl, -O-(C3-C 12 )-cycloalkyl, -S-(C1-C 12 )-alkyl, -S-(C3-C 12 )-cycloalkyl, -COO-(C1-C 12 )-alkyl, -COO-(C3-C 12 )-cycloalkyl, -CONH-(C1-C 12 )-alkyl, -CONH-(C3-C 12 )-cycloalkyl, -CO-(C1-C 12 )-alkyl, -CO-(C3-C 12 )-cycloalkyl, -N-[(C1-C 12 )-alkyl]2, -(C6-C 20 )-aryl, -(C6-C 20 )-aryl-(C1-C 12 )-alkyl, -(C6-C 20 )-aryl-O-(C1-C 12 )-alkyl, -(C3-C 20 )-heteroaryl, -(C3-C 20 )-heteroaryl-(C1-C 12 )-alkyl, -(C3-C 20 )-heteroaryl-O-(C1-C 12 )-alkyl, -(C1-C substituted with one or more substituents selected from -COOH, -OH, -SH, -SO3H, -CN, -NH2 or halogen 30 )-alkyl, -(C3-C 12 )-cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -(C6-C 20 )-aryl or -(C3-C 20)-heteroaryl group; X, Y and Z are each independently -O, -NH, -S, -N-(C1-C 30 )-alkyl or -(C1-C 30 )-aryl; [Chemical Structure] is selected from the group consisting of -CH((C1-C 30 )-alkyl))-, -(C=O)-, -CH(OH)-, -SO2-, -SO- and -CH(SOCH3)-; and R3 is optionally substituted -(C1-C 12 )-alkyl, -(C3-C 12 )-cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -O-(C1-C 12 )-alkyl, -O-(C1-C 12 )-alkyl-(C6-C 20 )-aryl, -O-(C3-C 12 )-cycloalkyl, -S-(C1-C 12 )-alkyl, -S-(C3-C 12 )-cycloalkyl, -COO-(C1-C 12 )-alkyl, -COO-(C3-C 12 )-cycloalkyl, -CONH-(C1-C 12 )-alkyl, -CONH-(C3-C 12 )-cycloalkyl, -CO-(C1-C 12 )-alkyl, -CO-(C3-C 12 )-cycloalkyl, -N-[(C1-C 12 )-alkyl]2, -(C6-C 20 )-aryl, -(C6-C 20 )-aryl-(C1-C 12 )-alkyl, -(C6-C 20 )-aryl-O-(C1-C 12 )-alkyl, -(C3-C 20 )-heteroaryl, -(C3-C 20 )-heteroaryl-(C1-C 12 )-alkyl, -(C3-C 20)-heteroaryl-O-(C1-C 12 -(C1-C) substituted with one or more substituents selected from )-alkyl, -COOH, -OH, -SH, -SO3H, -CN, -NH2 or halogens 30 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -(C6-C 20 )-aryl or-(C3-C 20 It is a heteroaryl group.

[0047] In any of the embodiments described herein, the Akt3 modulator is Equation III: [ka] A compound of or a pharmaceutically acceptable enantiomer, salt, or solvate thereof, where: R1 may be -(C1-C 12 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -O-(C1-C 12 )-alkyl,-O-(C1-C 12 )-alkyl-(C6-C 20 )-aryl, -O-(C3-C 12 )-Cycloalkyl, -S-(C1-C 12 )-alkyl,-S-(C3-C 12 )-Cycloalkyl, -COO-(C1-C 12 )-alkyl,-COO-(C3-C 12 )-Cycloalkyl, -CONH-(C1-C 12 )-alkyl,-CONH-(C3-C 12 )-Cycloalkyl, -CO-(C1-C 12 )-alkyl,-CO-(C3-C 12 )-Cycloalkyl, -N-[(C1-C 12 )-alkyl]2,-(C6-C 20 )-aryl,-(C6-C 20 )-aryl-(C1-C 12)-alkyl,-(C6-C 20 )-aryl-O-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl, -(C3-C 20 )-heteroaryl-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl-O-(C1-C 12 -(C1-C) substituted with one or more substituents selected from )-alkyl, -COOH, -OH, -SH, -SO3H, -CN, -NH2 or halogens 30 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -(C6-C 20 )-aryl or-(C3-C 20 )-heteroaryl group; X, Y, and Z are independently -O, -NH, -S, -N-(C1-C 30 )-alkyl or -(C1-C 30 )-Aryl; [ka] is -CH((C1-C 30 It is -alkyl))-, -(C=O)-, -CH(OH), -SO2-, -SO- or -CH(SOCH3)-; and R4 is -(C1-C 12 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -O-(C1-C 12 )-alkyl,-O-(C1-C 12 )-alkyl-(C6-C 20 )-aryl, -O-(C3-C 12 )-Cycloalkyl, -S-(C1-C 12 )-alkyl,-S-(C3-C 12 )-Cycloalkyl, -COO-(C1-C 12 )-alkyl,-COO-(C3-C 12 )-Cycloalkyl, -CONH-(C1-C12 )-alkyl,-CONH-(C3-C 12 )-Cycloalkyl, -CO-(C1-C 12 )-alkyl,-CO-(C3-C 12 )-Cycloalkyl, -N-[(C1-C 12 )-alkyl]2,-(C6-C 20 )-aryl,-(C6-C 20 )-aryl-(C1-C 12 )-alkyl,-(C6-C 20 )-aryl-O-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl, -(C3-C 20 )-heteroaryl-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl-O-(C1-C 12 It is either -alkyl, -COOH, -OH, -SH, -SO3H, -CN, -NH2, or halogen.

[0048] In any of the embodiments described herein, the Akt3 modulator is given by Equation IV: [ka] It is a compound of or a pharmaceutically acceptable enantiomer, salt, or solvate of the compound.

[0049] In any of the embodiments described herein, the method further includes administering a second therapeutic agent to a subject.

[0050] In any of the embodiments described herein, the second therapeutic agent is selected from the group consisting of nutritional supplements, chemotherapeutic agents, anti-inflammatory agents, immunosuppressants, cholinesterase inhibitors, antidepressants, anxiolytics, antipsychotics, riluzole, edaravone, dopamine agonists, MAO B inhibitors, catechol O-methyltransferase inhibitors, anticholinergics, anticonvulsants, tetrabenazine, carbidopa-levodopa, antispasmodics, antibodies, fusion proteins, enzymes, nucleic acids, ribonucleic acids, antiproliferative agents, cytotoxic agents, appetite stimulants, 5-HT3 antagonists, Cox-2 inhibitors, and combinations thereof.

[0051] In another embodiment, a method for treating cachexy in a subject requiring treatment is described, comprising administering to the subject a composition comprising a selective inhibitor of Akt3 in an effective amount that inhibits Akt3 signaling and activates adipogenesis in adipocytes.

[0052] In any of the embodiments described herein, the method further includes administering a second therapeutic agent to a subject.

[0053] In any of the embodiments described herein, the second therapeutic agent is selected from the group consisting of appetite stimulants, nutritional supplements, 5-HT3 antagonists, Cox-2 inhibitors, chemotherapeutic agents, anti-inflammatory agents, immunosuppressants, cholinesterase inhibitors, antidepressants, anxiolytics, antipsychotics, riluzole, edaravone, dopamine agonists, MAO B inhibitors, catechol O-methyltransferase inhibitors, anticholinergics, anticonvulsants, tetrabenazine, carbidopa-levodopa, antispasmodics, antibodies, fusion proteins, enzymes, nucleic acids, ribonucleic acids, antiproliferative agents, cytotoxic agents, and combinations thereof.

[0054] In any of the embodiments described herein, the second therapeutic agent is an appetite stimulant, a nutritional supplement, a 5-HT3 antagonist, or a Cox-2 inhibitor.

[0055] In any of the embodiments described herein, the subject is a neurodegenerative disease, cachexia, eating disorders, obesity complications, inflammatory diseases, virus-induced inflammatory responses, Gulf War syndrome, tuberous sclerosis, retinitis pigmentosa, graft rejection, cancer, or a combination thereof.

[0056] In any of the embodiments described herein, the Akt3 inhibitor is a compound selected from the group consisting of the following: [ka] [ka]

[0057] Detailed description of the invention I. Definition It should be recognized that the present invention is not limited to the compositions and methods and experimental conditions described herein, and that they may be modified. Since the scope of the present invention is limited only by the appended claims, it should also be understood that the terms used herein are for illustrative purposes only and are not intended to limit any particular embodiment.

[0058] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention pertains. Any composition, method, and substance similar to or equivalent to those described herein may be used in carrying out or testing this invention. All publications mentioned herein are incorporated herein by reference in their entirety.

[0059] The use of singular expressions in the description of the claimed invention herein (particularly in the claims) should be interpreted as encompassing both singular and plural forms unless otherwise specifically stated herein or unless it is clearly contrary to the context.

[0060] Unless otherwise specified herein, the ranges of values ​​described herein are intended as an abbreviation to refer simply to each individual value that falls within that range, and each separate value is incorporated herein to the same extent as if it were mentioned individually.

[0061] The use of the term "about" is intended to mean values ​​within a range of approximately ±10% above or below the stated value; in other embodiments, the value may be within a range of approximately ±5% above or below the stated value; in other embodiments, the value may be within a range of approximately ±2% above or below the stated value; in other embodiments, the value may be within a range of approximately ±1% above or below the stated value. The aforementioned ranges are intended to be apparent from the context and do not imply further limitation. All methods described herein may be performed in any appropriate order unless otherwise specifically stated or unless it is clearly contrary to the context. Any and all examples or illustrative terms provided herein (e.g., "such as") are intended solely to facilitate the understanding of the invention and do not impose limitations on the scope of the invention unless otherwise specifically stated. No term used herein should be construed as meaning that any element not described in any of the claims is essential for the practice of the invention.

[0062] As used herein, the terms “cancer” and, similarly, “tumor” refer to a condition in which abnormally replicating cells of host origin are present in a detectable amount in a subject. Cancer can be malignant or non-malignant. Cancer or tumors include, but are not limited to, adult T-cell leukemia / lymphoma (including those caused by human T-cell lymphotropic virus (HTLV-1)), cholangiocarcinoma; brain cancer; breast cancer; cervical cancer; choriocarcinoma; colon cancer; endometrial cancer; esophageal cancer; gastric (body) cancer; neoplasm in situ; leukemia; lymphoma; liver cancer; lung cancer (e.g., small cell and non-small cell); melanoma; neuroblastoma; oral cancer; ovarian cancer; pancreatic cancer; prostate cancer; rectal cancer; kidney cancer; sarcoma; skin cancer; testicular cancer; thyroid cancer; and other carcinomas and sarcomas. As used herein, the term “lymphoma” refers to cancers of the lymphatic system or blood cancers that develop from lymphocytes. Cancer can be primary or metastatic. Non-cancerous diseases may be associated with mutational alterations of elements of the Ras signaling pathway, and the compounds disclosed herein may be used to treat these non-cancerous diseases. Such non-cancerous diseases include neurofibromatosis; Leopard syndrome; Noonan syndrome; Regius syndrome; Costello syndrome; cardiac-facial-cutaneous syndromes; hereditary gingival fibromatosis type 1; autoimmune lymphoproliferative syndromes; and capillary malformations-arteriovenous malformations.

[0063] The term "stimulate expression" means, for example, inducing expression or activity, or influencing expression to induce increased / greater expression or activity compared to a normal, healthy control.

[0064] The terms “immunoactivating response,” “activated immune response,” and “immunostimulated response” refer to responses that initiate, induce, enhance, increase, or improve the efficiency of innate or adaptive immunity. Such immune responses include, for example, the development of beneficial humoral (antibody-mediated) and / or cellular (antigen-specific T cell or secretion product-mediated) responses to peptides in a recipient patient. Such responses may be active responses induced by immunogen administration or passive responses induced by antibody or sensitized T cell administration. Cellular immune responses are triggered by the presentation of polypeptide epitopes conjugated to class I or class II major histocompatibility complex ("MHC") molecules to activate antigen-specific CD4+ T helper cells and / or CD8+ cytotoxic T cells. Responses may also include the activation of monocytes, macrophages, natural killer (NK) cells, basophils, dendritic cells, astrocytes, microglia, eosinophils, neutrophils, or other components of innate immunity. The presence of a cell-mediated immunological response can be determined by a proliferation assay (CD4+ T cells) or a cytotoxic T lymphocyte ("CTL") assay. The relative contributions of humoral and cellular responses to the protective or therapeutic effect of an immunogen can be distinguished by separately isolating antibodies and T cells from immunized syngenes and measuring their protective or therapeutic effects in a second subject.

[0065] The terms "suppressive immune response" and "immunosuppressive response" refer to responses that reduce or prevent the activation or efficiency of innate or adaptive immunity.

[0066] The term "immune tolerance" as used herein refers to any mechanism that prevents, suppresses, or shifts a potentially harmful immune response to a non-harmful immune response (as collectively referred to here by Bach, et al., N. Eng. J. Med., 347:911-920 (2002)).

[0067] The term "tolerance vaccine" as used here refers to an antigen-specific treatment typically used to attenuate the autoreactive T and / or B-cell response while leaving overall immune function intact.

[0068] An "immunogenic agent" or "immunogen," when administered to mammals, can induce an immunological response, optionally in combination with an adjuvant.

[0069] The term "immune cells" as used herein refers to cells of the innate and adaptive immune systems, including lymphocytes such as neutrophils, eosinophils, basophils, monocytes, macrophages, dendritic cells, B cells, T cells, and NK cells.

[0070] The term "conventional T cells" as used herein refers to T lymphocytes that express the αβ T cell receptor (TCR) and its co-receptor, CD4 or CD8. Conventional T cells are found in peripheral blood, lymph nodes, and tissues. See Roberts and Girardi, “Conventional and Unconventional T Cells”, Clinical and Basic Immunodermatology, pp. 85-104, (Gaspari and Tyring (ed.)), Springer London (2008), which is incorporated herein by reference in its entirety.

[0071] The "atypical T cells" used here are lymphocytes that express the γδ TCR and can generally reside in epithelial environments such as the skin, gastrointestinal tract, or urogenital tract. Another subset of atypical T cells is invariant natural killer T (NKT) cells, which possess the phenotype and functional capabilities of typical T cells as well as the characteristics of natural killer cells (e.g., cytolytic activity). See the same literature.

[0072] The term "Treg" used here refers to one or more regulatory T cells. Regulatory T cells are a subpopulation of T cells that modulate the immune system, maintain tolerance to autoantigens, and suppress the immune stimulation or activation response of other cells. Regulatory T cells exist in many forms, but the best understood are those that express CD4, CD25, and Foxp3.

[0073] The terms "natural Treg" or "nTreg" used here refer to one or more regulatory T cells that develop in the thymus.

[0074] The terms "inducible Treg" or "iTreg" used here refer to one or more regulatory T cells that develop from mature CD4+ normal T cells outside the thymus.

[0075] The "biological activity" of Akt3 refers to the biological function of the Akt3 polypeptide. Biological activity can be increased or decreased by changes in the basal level activity of the polypeptide, changes in the basal level avidity of the polypeptide, the amount of polypeptide, the ratio of Akt3 to one or more other Akt isoforms (e.g., Akt1 or Akt2) proteins, changes in the polypeptide expression level (including changes in Akt3 mRNA expression), or a combination thereof. For example, a bioavailable Akt3 polypeptide is one that has kinase activity, can bind to an Akt3 substrate, and can be phosphorylated. A non-biavailable Akt3 polypeptide is one that is mislocalized or cannot bind to an Akt substrate and cannot be phosphorylated.

[0076] The terms "specifically binds" or "exhibits specific binding" to a target used herein refer to a binding reaction that determines the presence of the molecule in the presence of a heterogeneous population of other biologics.

[0077] Under specified immunoassay conditions, certain molecules preferentially bind to specific targets and do not bind in significant amounts to other biofactors present in the sample. Specific binding of an antibody to a target under such conditions requires the antibody to be selected for its specificity to that target. Various immunoassay formats can be used to select antibodies that are specifically immunoreactive with a particular protein. For example, enzyme-linked immunosorbent assay (ELISA) is routinely used to select monoclonal antibodies that specifically react with a certain protein. For example, see Harlow and Lane (1988), Antibodies, A Laboratory Manual, Cold Spring Harbor Publications, New York (included herein in whole by reference) for a description of immunoassay formats and conditions that may be used to determine specific immunoreactivity.

[0078] The terms “oligonucleotide” and “polynucleotide” generally refer to any polyribonucleotide or polydeoxyribonucleotide that may be unmodified RNA or DNA, or modified RNA or DNA. Therefore, as used herein, for example, “polynucleotide” refers, in particular, to single-stranded and double-stranded DNA, DNA which is a mixture of single-stranded and double-stranded regions, single-stranded and double-stranded RNA and RNA which is a mixture of single-stranded and double-stranded regions, and hybrid molecules of DNA and RNA which may be single-stranded or, more typically, double-stranded or a mixture of single-stranded and double-stranded regions. The terms “nucleic acid” or “nucleic acid sequence” also include the polynucleotides defined above.

[0079] Furthermore, the term "polynucleotide" as used here refers to a triple-stranded region containing RNA, DNA, or both. The strands in such a region may be from the same molecule or different molecules. A region may contain all of one or more molecules, but more typically it contains only the regions of some molecules. One of the molecules in a triple-helix region is often an oligonucleotide.

[0080] As used herein, the term "polynucleotide" includes the DNA or RNA described above that contains one or more modified bases. Therefore, DNA or RNA having a backbone modified for stability or other reasons is a "polynucleotide" as intended herein. Furthermore, DNA or RNA containing abnormal bases such as inosine or modified bases such as tritylated bases, as merely two examples, is a polynucleotide as used herein.

[0081] The term "polypeptide" as used herein refers to a chain of amino acids of any length, regardless of modifications (e.g., phosphorylation or glucosylation). The term "polypeptide" includes proteins and their fragments. Polypeptides can be "exogenous," meaning they are foreign to the host cell from which they are utilized, such as human polypeptides produced by bacterial cells. Polypeptides are described here as amino acid residue sequences. These sequences are written from left to right, from the amino terminus to the carboxyl terminus. According to standard nomenclature, amino acid residue sequences are represented by three-letter or one-letter abbreviations as follows: alanine (Ala, A), arginine (Arg, R), asparagine (Asn, N), aspartic acid (Asp, D), cysteine ​​(Cys, C), glutamine (Gln, Q), glutamic acid (Glu, E), glycine (Gly, G), histidine (His, H), isoleucine (Ile, I), leucine (Leu, L), lysine (Lys, K), methionine (Met, M), phenylalanine (Phe, F), proline (Pro, P), serine (Ser, S), threonine (Thr, T), tryptophan (Trp, W), tyrosine (Tyr, Y), and valine (Val, V).

[0082] A "variant" is a polypeptide or polynucleotide that differs from a control polypeptide or polynucleotide but retains essential properties. Typical polypeptide variants differ in amino acid sequence from other control polypeptides. Generally, the differences are limited such that the sequences of the control polypeptide and the variant are closely similar as a whole and identical in many regions. Variants and control polypeptides may differ in amino acid sequence due to one or more modifications (e.g., substitution, addition, and / or deletion). Substitutable or inserted amino acid residues may or may not be encoded by the genetic code. Polypeptide variants may be naturally occurring, such as allele variants, or they may be variants not known to exist naturally.

[0083] Modifications and alterations can be made to the structure of the polypeptide of the present invention, and a molecule with similar properties as a polypeptide can still be obtained (e.g., conservative amino acid substitution). For example, one amino acid can be substituted for another amino acid in the sequence without any noticeable loss of activity. Since the biological functional activity of a polypeptide is determined by its interactivity and properties, it is possible to make an amino acid sequence substitution in the polypeptide sequence and still obtain a polypeptide with similar properties.

[0084] To achieve this transformation, the hydrophobicity and hydrophilicity indices of amino acids can be considered. The importance of hydrophobicity and hydrophilicity amino acid indices in conferring interactive biological functions to polypeptides is generally understood in this field. It is known that by substituting one amino acid with another amino acid having a similar hydrophobicity or hydrophilicity index or score, polypeptides with similar biological activity can still be obtained. Each amino acid is assigned a hydrophobicity and hydrophilicity index based on its hydrophobicity and charge properties. These indicators are isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine / cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (-0.7); serine (-0.8); tryptophan (-0.9); tyrosine (-1.3); proline (-1.6); histidine (-3.2); glutamate (-3.5); glutamine (-3.5); aspartate (-3.5); asparagine (-3.5); lysine (-3.9); and arginine (-4.5).

[0085] The relative hydrophobic and hydrophilic index characteristics of amino acids are thought to determine the secondary structure of the resulting polypeptide, which in turn determines the interaction between the polypeptide and other molecules such as enzymes, substrates, receptors, antibodies, antigens, and cofactors. It is known in the art that functionally equivalent polypeptides can be obtained by substituting amino acids with other amino acids having similar hydrophobic and hydrophilic index characteristics. In such modifications, it is preferable that the amino acid substitutions result in hydrophobic and hydrophilic index characteristics within ±2, particularly preferably within ±1, and even more preferably within ±0.5.

[0086] Substitutions of similar amino acids may also be made based on hydrophilicity, particularly when the resulting biologically functional equivalent polypeptide or peptide is intended for use in immunological embodiments. The following hydrophilicity values ​​are assigned to amino acid residues: arginine (+3.0); lysine (+3.0); aspartate (+3.0±1); glutamate (+3.0±1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); proline (-0.5±1); threonine (-0.4); alanine (-0.5); histidine (-0.5); cysteine ​​(-1.0); methionine (-1.3); valine (-1.5); leucine (-1.8); isoleucine (-1.8); tyrosine (-2.3); phenylalanine (-2.5); and tryptophan (-3.4). It is understood that by substituting amino acids with others having similar hydrophilic values, it is possible to obtain a biologically equivalent, and in particular immunologically equivalent, polypeptide. In such modifications, preferably, the amino acid substitutions have hydrophilic values ​​within ±2, particularly preferably within ±1, and even more preferably within ±0.5.

[0087] As outlined above, amino acid substitutions are generally based on the relative similarity of amino acid side chain substituents, such as hydrophobicity, hydrophilicity, charge, and size. Examples of substitutions that take various of the above characteristics into account are well known to those skilled in the art and include (original residue: exemplary substitution): (Ala:Gly, Ser), (Arg:Lys), (Asn:Gln, His), (Asp:Glu,Cys, Ser), (Gln:Asn), (Glu:Asp), (Gly:Ala), (His:Asn, Gln), (Ile:Leu, Val), (Leu:Ile, Val), (Lys:Arg), (Met:Leu, Tyr), (Ser:Thr), (Thr:Ser), (Trp:Tyr), (Tyr:Trp, Phe), and (Val:Ile, Leu). The embodiments of the present invention are therefore intended to be functional or biological equivalents of the polypeptide described above. In particular, the embodiments of the polypeptide may include variants having about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with the polypeptide of interest.

[0088] The term “percent (%) sequence identity” is defined as the percentage of nucleotides or amino acids in a candidate sequence that are identical to a nucleotide or amino acid in a control nucleic acid sequence, after the sequences have been aligned and gaps introduced, if necessary, to achieve maximum percent sequence identity. Alignment for the purpose of determining percent sequence identity can be achieved by various methods within the scope of the art of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2, or Megalign (DNASTAR). Appropriate parameters for measuring alignment, including any algorithm necessary to achieve maximum alignment over the full length of the sequences being compared, can be determined by known methods.

[0089] For the purposes of this text, the % sequence identity of a nucleotide or amino acid sequence C to a nucleic acid sequence D (or, to put it another way, a sequence C that has or contains a certain % sequence identity to a sequence D) is calculated as follows: 100 x fraction W / Z (In the formula, W is the number of nucleotides or amino acids scored as identical matches in the alignment of C and D by the sequence alignment program, and Z is the total number of nucleotides or amino acids in D). When the length of sequence C is not equal to the length of sequence D, it is recognized that the % sequence identity of C to D is not equal to the % sequence identity of D to C.

[0090] The term "carrier" refers to a natural or synthetic, organic or inorganic component that is combined with an active ingredient to facilitate its application.

[0091] The term "pharmaceutically acceptable" refers to non-toxic materials that do not interfere with the effectiveness of the biological activity of the active ingredient.

[0092] The term "pharmaceutically acceptable carrier" means one or more suitable solid or liquid fillers, diluents, or encapsulants suitable for administration to humans or other vertebrates.

[0093] The term “effective dose” or “therapeutic effective dose” refers to a dosage sufficient to treat the disorder, disease, or condition being treated, or to otherwise provide the desired pharmacological and / or physiological effect. The exact dosage varies depending on a variety of factors, including subject-dependent variables (e.g., age, immune system health), the disease, and the treatment being performed.

[0094] The terms “individual,” “subject,” and “patient” are used interchangeably here and refer to mammals, including but not limited to humans, rodents such as mice and rats, and other laboratory animals.

[0095] The term "motor nerve" as used herein refers to a neuron whose cell body is located in the motor cortex, brainstem, or spinal cord, and whose axon projects into or outside the spinal cord to directly or indirectly control effector organs, primarily muscles and glands.

[0096] II. Methods for treating and preventing diseases by regulating Akt3 signaling Methods for treating and preventing diseases by modulating Akt3 signaling are disclosed herein. Non-exclusive examples of diseases include neurodegenerative diseases, cachexia, eating disorders, obesity complications, inflammatory diseases, virus-induced inflammatory responses, Gulf War Syndrome, tuberous sclerosis, retinitis pigmentosa, graft rejection, cancer, ischemic tissue injury, traumatic tissue injury, and combinations thereof. In some embodiments, the compounds disclosed herein modulate Akt3 signaling and may be used to treat various other diseases and disorders suspected to be caused by PI3K / Akt signaling dysfunction.

[0097] In one embodiment, the disclosed Akt3 inhibitor may be administered to subjects diagnosed with an eating disorder in an effective dose that promotes lipogenesis and reverses excessive weight loss.

[0098] Akt3 regulation for the treatment of neurodegenerative diseases One embodiment provides a method for treating or preventing neurodegenerative diseases in subjects requiring treatment, comprising administering to a subject a composition comprising an Akt3 modulator in an amount effective to modulate Akt3 signaling and treat or delay the progression of the disease. Non-limited examples of neurodegenerative diseases include Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, motor neuron disorders, Huntington's disease, HIV-induced neurodegeneration, Lewy body disease, spinal muscular atrophy, prion diseases, spinocerebellar ataxia, familial amyloid polyneuropathy, and combinations thereof.

[0099] Neurodegenerative diseases occur when nerve cells in the brain or peripheral nervous system lose function over time and eventually die. In many neurodegenerative diseases, chronic neuroinflammation contributes to disease progression. Current treatments help alleviate some of the physical or mental symptoms associated with neurodegenerative diseases, but there is currently no way to slow disease progression, and no cure is known.

[0100] While the mechanisms driving neurodegenerative processes remain largely unknown, growing evidence suggests a crucial role for immunity and the immune system in the pathogenesis of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, multiple sclerosis, spinal muscular atrophy, and amyotrophic lateral sclerosis (ALS). Regulatory T cells (Tregs) are a subset of CD4+ T cells that suppress the immune response and are essential mediators of self-tolerance and immune homeostasis (Sakaguchi, et al., Cell, 133, 775-787 (2008); incorporated herein by reference as a whole). Several pieces of evidence suggest that Tregs play a vital role in the progression of neurodegenerative diseases. Akt3 modulates the inhibitory function of native Tregs and the polarization of induced Tregs, and therefore, Akt3 regulation in immune cells has been shown to modulate the immune response. More specifically, Akt3 in activated immune cells induces an immunosuppressive response, while Akt3 inhibition in immune cells induces an immunosuppressive response. Although not bound by either theory, regulating Akt3 signaling in immune cells is thought to be useful for the treatment and prevention of neurodegenerative diseases.

[0101] One embodiment provides a method for treating or prophylacticizing Akt3 activators in subjects requiring treatment, by administering Akt3 activators to subjects in amounts effective in inducing an immunosuppressive response and treating or delaying the progression of a disease. In one embodiment, Akt3 activators modulate the immune response by increasing the suppressive function of immunosuppressive cells. In one embodiment, Akt3 is selectively activated in immune cells. Examples of immune cells include, but are not limited to, T cells, B cells, macrophages, and glial cells, such as astrocytes, microglia, and oligodendrocytes. In a preferred embodiment, Akt3 is activated in Tregs. Akt3 activators may also be used to increase or promote Treg activity or production, increase the production of cytokines such as IL-10 from Tregs, increase Treg differentiation, increase Treg number, or increase Treg survival.

[0102] Other embodiments provide methods for treating or preventing neurodegenerative diseases in subjects requiring treatment by inhibiting immunosuppressive responses and administering Akt3 inhibitors to subjects in amounts effective to treat or prevent disease progression. In some embodiments, Akt3 inhibitors modulate the immune response by reducing immunosuppressive responses or increasing immunostimulatory responses. In some embodiments, Akt3 is selectively inhibited in immune cells. Examples of immune cells include, but are not limited to, T cells, B cells, macrophages, and glial cells, such as astrocytes, microglia, and oligodendrocytes. In preferred embodiments, Akt3 is inhibited in Tregs.

[0103] 1. Subject to treatment a. Amyotrophic lateral sclerosis (ALS) In one embodiment, the disclosed Akt3 modulator can treat or prevent ALS. ALS, also known as Lou Gehrig's disease, is a progressive neurodegenerative disease affecting motor neurons in the brain and spinal cord. Symptoms of ALS include, but are not limited to, difficulty speaking, swallowing, walking, moving, and breathing. AL typically affects men and women between the ages of 40 and 70. There are two distinct types of ALS: sporadic and familial. Sporadic ALS, the most common form of the disease in the United States, accounts for 90–95 percent of all cases. Familial ALS is associated with mutations in Cu / Zn superoxide dismutase (SOD1). Oxidative stress, mitochondrial dysfunction, excitotoxicity, protein aggregation, endoplasmic reticulum stress, axonal transport dysfunction, dysregulation of neuron-glial interactions, and apoptosis have all been shown to contribute to motor neuron damage in the presence of mutant OD1.

[0104] While not bound by any single theory, it is believed that Treg dysfunction plays a role in the progression of ALS, and that Akt3 modulators may be able to treat or prevent the progression of ALS. Some subjects with rapidly progressing ALS have been found to have a deficiency in the Treg master transcription factor FOXP3, which leads to dysfunction of Treg inhibitory function. One embodiment provides a method for treating ALS in subjects in need, comprising administering an Akt3 activator to a subject in need in an effective amount that activates Akt3 in immune cells and induces an immunosuppressive response. In a preferred embodiment, Akt3 is activated in Tregs.

[0105] In one embodiment, administration of an Akt3 activator to a subject with ALS slows disease progression and prolongs the subject's survival.

[0106] Other motor neurological disorders that may be treated or prevented using the disclosed Akt3 activator include, but are not limited to, progressive bulbar palsy, pseudobulbar palsy, primary lateral sclerosis, spinal muscular atrophy, and post-polio syndrome.

[0107] b. Parkinson's disease Parkinson's disease is a neurodegenerative disorder that primarily affects dopamine-producing neurons in a specific area of ​​the brain called the substantia nigra. Parkinson's disease is a progressive disorder that worsens over time due to the increasing number of damaged or dying neurons. The cause of neuronal cell death in Parkinson's disease is unknown. Symptoms of Parkinson's disease include, but are not limited to, tremors of the hands, arms, legs, jaw, or head; stiffness of the limbs and torso; bradykinesia (slowness of movement); and impairment of balance and coordination.

[0108] One embodiment provides a method for treating Parkinson's disease, comprising administering an Akt3 modulator to a subject in need in an effective amount that activates or inhibits Akt3 in immune cells and induces an immunosuppressive response. In one embodiment, administration of the Akt3 activator to a subject with Parkinson's disease slows or halts disease progression to unaffected areas of the brain.

[0109] In one embodiment, the disclosed Akt3 activator may be administered prophylactically to a subject if the subject has a family history of Parkinson's disease or other neurodegenerative disease. The Akt3 activator protects neurons from disease induction or slows disease induction.

[0110] c. Huntington's disease Huntington's disease is a progressive neurodegenerative disease characterized by the progressive decay of nerve cells in the brain. Symptoms of Huntington's disease include, but are not limited to, involuntary motor problems and dysfunctions in voluntary movements, e.g., involuntary spasms, muscle rigidity, oculomotor hypertrichosis or abnormalities, gait, posture and balance disorders, difficulty with the physical output of speech or swallowing; cognitive impairments, e.g., difficulty organizing, prioritizing or consolidating tasks, lack of flexibility or tendency to stick to thoughts, actions or activities, lack of impulse control, lack of awareness of one's own actions and abilities, slowness of thinking or word-finding and difficulty learning new information; and mental disorders, e.g., depression. In one embodiment, the disclosed Akt3 modulator reduces or slows the progression of Huntington's disease.

[0111] One embodiment provides a method for treating Huntington's disease in a subject requiring it, by administering an Akt3 modulator in an effective amount that activates or inhibits Akt3 in immune cells and induces an immunosuppressive response. In one embodiment, the Akt3 modulator slows or halts the progression of disease symptoms in a subject with Huntington's disease. In another embodiment, the Akt3 modulator can alter the Treg / Th17 equilibrium.

[0112] Huntington's disease is largely hereditary; all children of parents with Huntington's disease have a 50 / 50 chance of inheriting the disease. In one embodiment, subjects with a family history of Huntington's disease may be prophylactically administered the disclosed Akt3 modulator before the onset of disease symptoms to prevent or slow the manifestation of disease symptoms.

[0113] d. Alzheimer's disease Alzheimer's disease is a progressive disorder that causes degeneration and ultimately cell death of brain cells. It is the leading cause of dementia—a continuous decline in thinking, behavior, and social skills that impairs a person's ability to function independently. Symptoms of Alzheimer's disease include, but are not limited to, memory loss, impaired thinking and reasoning abilities, difficulty in making judgments and decisions, and changes in personality and behavior. While the exact cause of Alzheimer's disease is not fully understood, the core problem is thought to be dysfunction in brain proteins that interfere with neuronal function, unleashing a series of toxic events. Damage most often begins in areas of the brain that control memory, but this process begins several years before the first symptoms appear. Neuronal loss spreads to other areas of the brain in a somewhat predictable pattern. In the later stages of the disease, the brain undergoes significant atrophy. Beta-amyloid plaques and tau protein concentrates most frequently contribute to the majority of neuronal damage and dysfunction in Alzheimer's disease.

[0114] One embodiment provides a method for treating Alzheimer's disease in a subject by administering an effective dose of an Akt3 activator to the subject, which hyperpolarizes Akt3 in Tregs and activates downstream neuroprotective pathways in the brain. In another embodiment, the subject is administered an effective dose of an Akt3 activator that reduces or eliminates the symptoms of Alzheimer's disease or slows disease progression.

[0115] Other embodiments provide a method for treating or preventing the progression of Alzheimer's disease in a subject by administering an Akt3 inhibitor to the subject in an effective dose that inhibits Akt3 in Tregs and induces an immune response or reduces an immunosuppressive response. In one embodiment, inhibition of Akt3 in Tregs leads to beta-amyloid plaque clearance, mitigation of neuroinflammatory responses, and recovery of cognitive decline.

[0116] In one embodiment, prophylactic administration of an Akt3 modulator to subjects with a family history of Alzheimer's disease can prevent or slow the onset of Alzheimer's disease.

[0117] e. Spinal muscular atrophy Spinal muscular atrophy ("SMA") is a group of chronic neuromuscular diseases characterized by progressive loss of motor nerves and muscle wasting. SMA is generally classified into four types, differing in severity and the life stage at which the disease manifests. These types are: SMA1 or Werdnig-Hoffmann disease ("pediatric" SMA) that manifests between 0 and 6 months of age; SMA2 or Dubovitz disease ("intermediate" SMA) that manifests between 6 and 18 months of age; SMA3 or Kugelberg-Wellander disease ("juvenile" SMA) that manifests after the age of one; and SMA4, which develops during adulthood ("adult-onset" SMA). The most severe form of SMA1 is sometimes referred to as SMA0 ("severe childhood" SMA). Signs and symptoms of SMA vary by type, but most commonly include, but are not limited to, body instability or a tendency to fall, difficulty sitting, standing, or walking, loss of strength in respiratory muscles, spasms, and difficulty eating and swallowing. All types of SMA are associated with exon deletions and / or point mutations in the SMN1 gene, which block the expression of the SMN protein. Depending on the type, SMA can be treated with various gene therapies, nutrition and respiratory support, orthopedics, and combinations thereof. Neuroprotective drugs are promising as a way to stabilize motor nerve loss, but currently available candidates have not yet achieved full progress in clinical trials. Therefore, further candidate neuroprotective drugs are needed for the treatment of SMA.

[0118] One embodiment provides a method for treating SMA in a subject by administering an effective amount of Akt3 activator to the subject that allows for motor neuron survival. In another embodiment, the subject is administered an effective amount of Akt3 modulator that reduces or eliminates the symptoms of SMA or slows disease progression.

[0119] Akt3 inhibition for the treatment of excessive weight loss Methods for treating or preventing excessive weight loss associated with diseases and disorders such as cachexia and eating disorders are disclosed herein. Exemplary methods include inhibiting Akt3 in subjects requiring it. While not bound by any one theory, Akt3 is thought to play a crucial role in adipogenesis. White adipogenesis requires activation of a transcriptional cascade involving the sequential induction of several transcription factors, including but not limited to FOXO1, several members of the C / EBP family, and PPARγ. FOXO1 is an essential negative regulator of adipogenesis and is primarily regulated by multi-residue phosphorylation / acetylation by enzymes including Akt. FOXO1 can also be regulated by the serine / threonine protein kinase SGK1. SGK1 is downstream of PI3K and can inhibit FOXO1 by phosphorylation. SGK1 is regulated by the serine / threonine protein kinase WNK1, which can also be regulated by Akt and SGK1. Akt3 inhibits adipogenesis through WNK1 phosphorylation, leading to downregulation of SGK1 activity and SGK-1-mediated inhibition of FOXO1. In one embodiment, inhibition of Akt3 in Tregs can promote adipogenesis and reverse disease derivative weight loss.

[0120] 1. Subject to treatment a. cachexy Cachexy, or wasting syndrome, is a multifactorial syndrome characterized by progressive skeletal muscle loss that cannot be fully recovered with conventional nutritional support and leads to progressive dysfunction. Cachexy is destructive to the extent that the body utilizes other energy sources, namely skeletal muscle and adipose tissue, when it senses nutritional deficiency. It is associated with the ability to fight infection, treatment tolerance, response to treatment, quality of life, and life expectancy.

[0121] In one embodiment, cachexy is caused by chronic diseases such as, but not limited to, AIDS, celiac disease, chronic obstructive pulmonary disease, multiple sclerosis, rheumatoid arthritis, congestive heart failure, tuberculosis, familial amyloid polyneuropathy, Crohn's disease, untreated and severe type 1 diabetes, anorexia nervosa, hyperthyroidism, and hormone deficiencies.

[0122] One embodiment provides a method for treating cachexia in a subject requiring treatment by administering an Akt3 inhibitor in an amount effective in reducing the symptoms of cachexia. Another embodiment provides a method for promoting weight gain in a subject requiring treatment by administering an Akt3 inhibitor in an amount effective in promoting adipogenesis in the subject. In one embodiment, the compounds disclosed herein are used to treat cachexia by modulating Akt3 rather than by modulating T regulatory cells.

[0123] In one embodiment, an Akt3 inhibitor may be prophylactically administered to subjects suspected of being susceptible to cachexia (for example, subjects diagnosed with other diseases) to prevent or slow the onset of cachexia syndrome.

[0124] b. Eating disorders Anorexia nervosa is an eating disorder characterized by weight loss or, in growing children, failure to gain weight, difficulty maintaining a weight appropriate for height, age, and stature, and often a distorted body image. One of the primary goals of treating eating disorders is the restoration of normal weight. In some embodiments, the compounds disclosed herein by formula I inhibit Akt3, which is hyperactivated by elevated estradiol levels in subjects with eating disorders. In some embodiments, the compounds disclosed herein by formula I may be used in the treatment of eating disorders.

[0125] Akt3 regulation for the treatment of obesity and its complications In one embodiment, the compounds disclosed herein that modulate Akt3 are used for the treatment of obesity and / or complications associated with obesity. In one embodiment, complications of obesity are selected from the group consisting of glucose intolerance, fatty liver, dyslipidemia, and combinations thereof. In one embodiment, the compounds disclosed herein are used to treat obesity and / or complications of obesity via modulation of Akt3, rather than via modulation of T regulatory cells.

[0126] Akt3 modulation for the treatment of inflammatory diseases Akt3 signal transduction is associated with chronic or acute inflammation that contributes to inflammatory diseases. One embodiment provides a method of treating or preventing an inflammatory disease in a subject in need thereof, comprising administering to the subject a composition comprising an Akt3 modulator in an effective amount to modulate Akt3 signal transduction and treat the disease or delay the progression thereof. In one embodiment, the Akt3 modulator activates Akt3 signal transduction and / or increases Treg activity or production, resulting in an immunosuppressive effect.

[0127] Non-limiting examples of inflammatory diseases include atopic dermatitis, allergy, asthma, and combinations thereof.

[0128] Akt3 modulation for the treatment of virus-induced inflammatory responses Akt3 signal transduction is associated with acute immune responses resulting from virus-induced inflammatory diseases such as severe acute respiratory syndrome ("SARS") and coronavirus disease 2019 ("COVID-19"). Therefore, in one embodiment, a method of treating a virus-induced inflammatory disease in a subject in need thereof comprises administering to the subject an Akt3 modulator in an effective amount to reverse or slow disease progression.

[0129] Akt3 modulation for the treatment of cancer In one embodiment, there is provided a method of treating or preventing cancer in a subject in need thereof, comprising modulating Akt3 signaling by administering to the subject an effective amount of a compound disclosed herein. In one embodiment, the compound disclosed herein inhibits Akt3 signaling and / or reduces Treg activity or production, and produces an immunoresponse activating effect.

[0130] In one embodiment, the cancer is selected from the group consisting of adult T-cell leukemia / lymphoma, bladder cancer, brain cancer, breast cancer, cervical cancer, colorectal cancer, esophageal cancer, kidney cancer, liver cancer, lung cancer, nasopharyngeal cancer, pancreatic cancer, prostate cancer, skin cancer, stomach cancer, uterine cancer, ovarian cancer, testicular cancer, and combinations thereof.

[0131] In one embodiment, the compounds and compositions disclosed herein are useful for treating leukemia. In one embodiment, the compounds and compositions disclosed herein that inhibit Akt3 are useful for treating leukemia. In these embodiments, the compounds and compositions disclosed herein that inhibit Akt3 are useful in vivo and ex vivo as immune response-stimulating therapy. The ability to inhibit Akt3 and thereby inhibit or reduce Treg-mediated immunosuppression allows a more robust immune response. In one embodiment, the compounds and compositions disclosed herein are also useful for stimulating or enhancing an immune stimulation or activation response involving T cells. In one embodiment, the compounds and compositions disclosed herein are useful for stimulating or enhancing an immune response in a host for treating leukemia by selectively inhibiting Akt3. In these embodiments, the compounds and compositions disclosed herein may be administered to the subject in an effective amount to stimulate T cells in the subject. Types of leukemia that may be treated with the compounds and compositions disclosed herein include, but are not limited to, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), adult T-cell leukemia / lymphoma (ATLL), and chronic myelomonocytic leukemia (CMML).

[0132] In one embodiment, ATLL is diagnosed almost exclusively in adults, with a median age of mid-60s. In one embodiment, there are four types of ATLL: (1) acute, (2) chronic, (3) smoldering, and (4) lymphomatous. In one embodiment, acute ATLL is the most common form, characterized by high white blood cell count, hypercalcemia, organ enlargement, and high lactose dehydrogenase. In one embodiment, lymphomatous ATLL manifests in the lymph nodes and accounts for less than 1% of circulating lymphocytes. In one embodiment, chronic and smoldering ATLL are characterized by a less aggressive clinical course and the possibility of long-term survival. In one embodiment, the 4-year survival rate for acute and lymphomatous ATLL is less than 5%. In one embodiment, the 4-year survival rates for chronic and smoldering forms of ATLL are 26.9% and 62%, respectively. In one embodiment, adult T-cell leukemia / lymphoma is caused by human T-cell lymphotropic virus (HTLV-1).

[0133] In one embodiment, the compounds and compositions disclosed herein are useful for treating ATLL. In one embodiment, the compounds and compositions disclosed herein that inhibit Akt3 are useful for treating ATLL. In one embodiment, Tregs expressing CD25 and FoxP3 become cancerous in ATLL cells. In one embodiment, ATLL cells exhibit an activated helper / inducer T cell phenotype but show strong immunosuppressive activity. In one embodiment, the compounds and compositions disclosed herein that inhibit Akt3 reduce the immunosuppressive response of ATLL cells. In another embodiment, the compounds and compositions disclosed herein that inhibit Akt3 enhance the immune-stimulating response and overcome the strong immunosuppressive activity of ATLL cells.

[0134] In some embodiments, the compounds and compositions disclosed herein, useful for the treatment of leukemia or ATLL, reduce or inhibit immunosuppressive responses, including but not limited to the immunosuppressive function of innate Treg (nTreg) cells and the induction of conventional T cells into inducible Treg (iTreg) cells. In these embodiments, the immunosuppressive function in nTreg cells that is reduced or inhibited is the secretion of one or more anti-inflammatory cytokines, such as, but not limited to, IL10, TGFβ, or a combination thereof. In some embodiments, a method for treating leukemia or adult T-cell leukemia / lymphoma comprises administering to the subject a secondary activator, such as, but not limited to, an anti-nausea drug, a chemotherapy drug, or an enhancer (e.g., cyclophosphamide).

[0135] Other indications In one embodiment, the compounds disclosed herein modulate Akt3 and are used to treat Gulf War Syndrome, tuberous sclerosis, retinitis pigmentosa, or graft rejection. In one embodiment, graft rejection is graft-versus-host disease. In one embodiment, the compounds disclosed herein are used to treat retinitis pigmentosa by modulating Akt3, rather than by modulating T regulatory cells. In one embodiment, the compounds disclosed herein are used to treat ischemic tissue injury or traumatic tissue injury. In one embodiment, ischemic tissue injury or traumatic tissue injury is ischemic tissue injury or traumatic tissue injury of the brain.

[0136] How to adjust Akt3 Akt3, also known as RAC-gammaserine / threonine-protein kinase, is an enzyme encoded by the Akt3 gene in humans. Akt kinases are regulators of insulin and growth factor-responsive cell signaling and are known to be involved in a wide range of biological processes, including but not limited to cell proliferation, differentiation, apoptosis, tumorigenesis, glycogen synthesis, and glucose uptake. Akt3 has been shown to be stimulated by platelet-derived growth factor ("PDGF"), insulin, and insulin-like growth factor 1 ("IGF1").

[0137] Akt3 kinase activity mediates serine and / or threonine phosphorylation of a range of downstream substrates. The nucleic acid sequence of Akt3 is known in this field. For example, see Genbank accession no. AF124141.1: Homo sapiens protein kinase B gamma mRNA, complete cds, which includes the entire sequence by reference: AGGGGAGTCATCATGAGCGATGTTACCATTGTGAAGGAAGGTTGGGTTCAGAAGAGGGGA GAATATATAAAAAACTGGAGGCCAAGATACTTCCTTTTGAAGACAGATGGCTCATTCATA GGATATAAAGAGAAACCTCAAGATGTGGATTTACCTTATCCCCTCAACAACTTTTCAGTG GCAAAAATGCCAGTTAATGAAAACAGAACGACCAAAGCCAAACACATTTATAATCAGATGT CTCCAGTGGACTACTGTTATAGAGAGAACATTTCATGTAGATACTCCAGAGGAAAGGGAA GAATGGACAGAAGCTATCCAGGCTGTAGCAGACAGACTGCAGAGGCAAGAAGAGGAGAGA ATGAATTGTAGTCCAACTTCACAAATTGATAATATAGGAGAGGAAGAGATGGATGCCTCT ACAACCCATCATAAAAGAAAGACAATGAATGATTTTGACTATTTGAAACTACTAGGTAAA GGCACTTTTGGGAAAGTTATTTTGGTTCGAGAGAAGGCAAGTGGAAAATACTATGCTATG AAGATTCTGAAGAAAGAAGTCATTATTGCAAAGGATGAAGTGGCACACACTCTAACTGAA AGCAGAGTATTAAAAGAACTAGACATCCCTTTTTAACATCCTTGAAATATTCCTTCCAG ACAAAAGACCGTTTGTGTTTTGTGATGGAATATGTTAATGGGGGCGAGCTGTTTTTCCAT TTGTCGAGAGAGCGGGTGTTCTCTGAGGACCGCACACGTTTCTATGGTGCAGAAATTGTC TCTGCCTTGGACTATCTACATTCCGGAAAGATTGTGTACCGTGATCTCAAGTTGGAGAAT CTAATGCTGGACAAAGATGGCCACATAAAAAATTACAGATTTTGGACTTTGCAAAGAAGGG ATCACAGATGCAGCCACCATGAAGACATTCTGTGGCACTCCAGAATATCTGGCACCAGAG GTGTTAGAAGATAATGACTATGGCCGAGCAGTAGACTGGTGGGGCTAGGGGGTTGTCATG TATGAAATGATGTGTGGGAGGTTACCTTTCTACAAACCAGGACCATGAGAAACTTTTTGAA TTAATATTAATGGAAGACATTAAATTTCCTCGAACACTCTCTTCAGATGCAAAATCATTG CTTTCAGGGCTCTTGATAAAGGATCCAAATAAACGCCTTGGTGGAGGACCAGATGATGCA AAAGAAATTATGAGACACAGTTTCTTCTCTGGAGTAAACTGGCAAGATGTATATGATAAA AAGCTTGTACCTCCTTTTAAACCTCAAGTAACATCTGAGACAGATACTAGATATTTTGAT GAAGAATTTACAGCTCAGACTATTACAATAACACCACCTGAAAAATATGATGAGGATGGT ATGGACTGCATGGACAATGAGAGGCGGCCGCATTTCCCTCAATTTTCCTACTCTGCAAGT GGACGAGAATAAGTCTCTTTCATTCTGCTACTTCACTGTCATCTTCAATTTATTACTGAA AATGATTCCTGGACATCACCAGTCCTAGCTCTTACACATAGCAGGGGCACCTTCCGACAT CCCAGACCAGCCAAGGGTCCTCACCCCTCGCCACCTTTCACCCTCATGAAAACACACATA CACGCAAATACACTCCAGTTTTTGTTTTTGCATGAAATTGTATCTCAGTCTAAGGTCTCA TGCTGTTGCTGCTACTGTCTTACTATTA (SEQ ID NO: 1).

[0138] Amino acid sequences are also known in the art. See, for example, UniProtKB / Swiss-Prot accession no. Q9Y243 (Akt3_HUMAN), which is incorporated by reference in its entirety herein and provides the following amino acid sequence: MSDVTIVKEGWVQKRGEYIKNWRPRYFLLKTDGSFIGYKEKPQDVDLPYPLNNFSVAKCQ LMKTERPKPNTFIIRCLQWTTVIERTFHVDTPEEREEWTEAIQAVADRLQRQEEERMNCS PTSQIDNIGEEEMDASTTHHKRKTMNDFDYLKLLGKGTFGKVILVREKASGKYYAMKILK KEVIIAKDEVAHTLTESRVLKNTRHPFLTSLKYSFQTKDRLCFVMEYVNGGELFFHLSRE RVFSEDRTRFYGAEIVSALDYLHSGKIVYRDLKLENLMLDKDGHIKITDFGLCKEGITDA ATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYEMMCGRLPFYNQDHEKLFELILM EDIKFPRTLSSDAKSLLSGLLIKDPNKRLGGGPDDAKEIMRHSFFSGVNWQDVYDKKLVP PFKPQVTSETDTRYFDEEFTAQTITITPPEKYDEDGMDCMDNERRPHFPQFSYSASGRE (Sequence ID 2).

[0139] The domain structure of Akt3 was reviewed in Romano, Scientifica, Volume 2013 (2013), Article ID 317186, p. 12 (incorporated herein by reference), and includes an N-terminal plextrin homology domain (PH), followed by a catalytic kinase domain (KD) and a C-terminal regulatory hydrophobic region. Both the catalytic and regulatory domains are important for the biological actions mediated by Akt protein kinases and exhibit the highest degree of homology among the 3Akt isoforms. The PH domain binds to lipid substrates such as phosphatidylinositol (3,4) diphosphate (PIP2) and phosphatidylinositol (3,4,5) triphosphate (PIP3). The ATP-binding site is located almost in the center of the catalytic kinase domain and exhibits a considerable degree of homology with other elements of the AGC kinase family, such as p70 S6 kinase (S6K), p90 ribosomal 6 kinase (RSK), protein kinase A (PKA), and protein kinase B (PKB). The hydrophobic regulatory moiety is a typical characteristic of the AGC kinase family. Sequence ID No. 2, Akt 3 is generally thought to have the molecular processing and domain structure outlined below. [Table 1]

[0140] The initiation methionine in SEQ ID NO: 2 is disposable in Akt3 function. Therefore, in one embodiment, the compound directly or indirectly regulates the expression or bioavailability of Akt3 having the following amino acid sequence: SDVTIVKEGWVQKRGEYIKNWRPRYFLLKTDGSFIGYKEKPQDVDLPYPLNNFSVAKCQ LMKTERPKPNTFIIRCLQWTTVIERTFHVDTPEEREEWTEAIQAVADRLQRQEEERMNCS PTSQIDNIGEEEMDASTTHHKRKTMNDFDYLKLLGKGTFGKVILVREKASGKYYAMKILK KEVIIAKDEVAHTLTESRVLKNTRHPFLTSLKYSFQTKDRLCFVMEYVNGGELFFHLSRE RVFSEDRTRFYGAEIVSALDYLHSGKIVYRDLKLENLMLDKDGHIKITDFGLCKEGITDA ATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYEMMCGRLPFYNQDHEKLFELILM EDIKFPRTLSSDAKSLLSGLLIKDPNKRLGGGPDDAKEIMRHSFFSGVNWQDVYDKKLVP PFKPQVTSETDTRYFDEEFTAQTITITPPEKYDEDGMDCMDNERRPHFPQFSYSASGRE (Sequence ID 3).

[0141] Two specific sites, one in the kinase domain (Thr-305, see SEQ ID NO: 2) and the other in the C-terminal regulatory region (Ser-472, see SEQ ID NO: 2), need to be phosphorylated for Akt3 to be fully activated. The interaction between the PH domain of Akt3 and TCL1A enhances Akt3 phosphorylation and activation. IGF-1 leads to the activation of Akt3, which may play a role in regulating cell survival.

[0142] Compositions for selectively modulating Akt3 activity and methods for using them are disclosed herein. Methods for using Akt3 modulators disclosed for the treatment or prevention of various diseases are also described.

[0143] A. Akt3 activator compound A composition for selectively activating Akt3 is provided herein. An example of an Akt3 activator is described in international application PCT / US2018 / 49715 (which is incorporated herein by reference in its entirety) and is described below.

[0144] One embodiment is given by formula I: [ka] We provide compounds of or pharmaceutically acceptable enantiomers, salts, or solvates thereof, where: Rings A, B, and C are independently six-membered aryl or N-containing heteroaryl monocycles or bicyclic ring systems containing zero or more N atoms, such as phenyl, pyridine, pyrimidine, pyridazine, pyrazine, triazine, quinoline, quinazoline, isoquinoline, naphthalene, naphthyridine, indole, isoindole, cinnoline, phthalazine, quinoxaline, pteridine, purine, and benzimidazole; R1 may be -(C1-C 12 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -O-(C1-C 12 )-alkyl,-O-(C1-C 12 )-alkyl-(C6-C 20 )-aryl, -O-(C3-C 12 )-Cycloalkyl, -S-(C1-C 12 )-alkyl,-S-(C3-C 12 )-Cycloalkyl, -COO-(C1-C 12 )-alkyl,-COO-(C3-C 12 )-Cycloalkyl, -CONH-(C1-C 12 )-alkyl,-CONH-(C3-C 12 )-Cycloalkyl, -CO-(C1-C 12 )-alkyl,-CO-(C3-C 12 )-Cycloalkyl, -N-[(C1-C 12 )-alkyl]2,-(C6-C 20 )-aryl,-(C6-C20 )-aryl-(C1-C 12 )-alkyl,-(C6-C 20 )-aryl-O-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl, -(C3-C 20 )-heteroaryl-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl-O-(C1-C 12 -(C1-C) is substituted with one or more substituents selected from alkyl, -COOH, -OH, -SH, -SO3H, -CN, -NH2 or halogens. 30 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -(C6-C 20 )-aryl or-(C3-C 20 Selected from )-heteroaryl groups; X, Y, and Z are independently = O, -NH, -S, -N-(C1-C) 30 )-alkyl or -(C1-C 30 ) - Selected from the arrows; [ka] is -CH((C1-C 30 It is -alkyl))-, -(C=O)-, -CH(OH), -SO2-, -SO- or -CH(SOCH3)-; and R3 may be -(C1-C 12 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -O-(C1-C 12 )-alkyl,-O-(C1-C 12 )-alkyl-(C6-C 20 )-aryl, -O-(C3-C 12 )-Cycloalkyl, -S-(C1-C 12 )-alkyl,-S-(C3-C 12 )-Cycloalkyl, -COO-(C1-C 12)-alkyl,-COO-(C3-C 12 )-Cycloalkyl, -CONH-(C1-C 12 )-alkyl,-CONH-(C3-C 12 )-Cycloalkyl, -CO-(C1-C 12 )-alkyl,-CO-(C3-C 12 )-Cycloalkyl, -N-[(C1-C 12 )-alkyl]2,-(C6-C 20 )-aryl,-(C6-C 20 )-aryl-(C1-C 12 )-alkyl,-(C6-C 20 )-aryl-O-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl, -(C3-C 20 )-heteroaryl-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl-O-(C1-C 12 -(C1-C) is substituted with one or more substituents selected from alkyl, -COOH, -OH, -SH, -SO3H, -CN, -NH2 or halogens. 30 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -(C6-C 20 )-aryl or-(C3-C 20 Selected from )-heteroaryl groups.

[0145] Other embodiments are given by formula II: [ka] We provide compounds of or pharmaceutically acceptable enantiomers, salts, or solvates thereof, where: R1 may be -(C1-C 12 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -O-(C1-C 12 )-alkyl,-O-(C1-C 12)-alkyl-(C6-C 20 )-aryl, -O-(C3-C 12 )-Cycloalkyl, -S-(C1-C 12 )-alkyl,-S-(C3-C 12 )-Cycloalkyl, -COO-(C1-C 12 )-alkyl,-COO-(C3-C 12 )-Cycloalkyl, -CONH-(C1-C 12 )-alkyl,-CONH-(C3-C 12 )-Cycloalkyl, -CO-(C1-C 12 )-alkyl,-CO-(C3-C 12 )-Cycloalkyl, -N-[(C1-C 12 )-alkyl]2,-(C6-C 20 )-aryl,-(C6-C 20 )-aryl-(C1-C 12 )-alkyl,-(C6-C 20 )-aryl-O-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl, -(C3-C 20 )-heteroaryl-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl-O-(C1-C 12 -(C1-C) is substituted with one or more substituents selected from alkyl, -COOH, -OH, -SH, -SO3H, -CN, -NH2 or halogens. 30 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -(C6-C 20 )-aryl or-(C3-C 20 Selected from )-heteroaryl groups; X, Y, and Z are independently -O, -NH, -S, -N-(C1-C 30 )-alkyl or -(C1-C 30 ) - Selected from the arrows; [ka] is -CH((C1-C30 It is -alkyl))-, -(C=O)-, -CH(OH), -SO2-, -SO- or -CH(SOCH3)-; and R3 may be -(C1-C 12 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -O-(C1-C 12 )-alkyl,-O-(C1-C 12 )-alkyl-(C6-C 20 )-aryl, -O-(C3-C 12 )-Cycloalkyl, -S-(C1-C 12 )-alkyl,-S-(C3-C 12 )-Cycloalkyl, -COO-(C1-C 12 )-alkyl,-COO-(C3-C 12 )-Cycloalkyl, -CONH-(C1-C 12 )-alkyl,-CONH-(C3-C 12 )-Cycloalkyl, -CO-(C1-C 12 )-alkyl,-CO-(C3-C 12 )-Cycloalkyl, -N-[(C1-C 12 )-alkyl]2,-(C6-C 20 )-aryl,-(C6-C 20 )-aryl-(C1-C 12 )-alkyl,-(C6-C 20 )-aryl-O-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl, -(C3-C 20 )-heteroaryl-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl-O-(C1-C 12 -(C1-C) is substituted with one or more substituents selected from alkyl, -COOH, -OH, -SH, -SO3H, -CN, -NH2 or halogens. 30 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -(C6-C 20)-aryl or-(C3-C 20 Selected from )-heteroaryl groups.

[0146] Other embodiments are given by formula III: [ka] We provide compounds of or pharmaceutically acceptable enantiomers, salts, or solvates thereof, where: R1 may be -(C1-C 12 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -O-(C1-C 12 )-alkyl,-O-(C1-C 12 )-alkyl-(C6-C 20 )-aryl, -O-(C3-C 12 )-Cycloalkyl, -S-(C1-C 12 )-alkyl,-S-(C3-C 12 )-Cycloalkyl, -COO-(C1-C 12 )-alkyl,-COO-(C3-C 12 )-Cycloalkyl, -CONH-(C1-C 12 )-alkyl,-CONH-(C3-C 12 )-Cycloalkyl, -CO-(C1-C 12 )-alkyl,-CO-(C3-C 12 )-Cycloalkyl, -N-[(C1-C 12 )-alkyl]2,-(C6-C 20 )-aryl,-(C6-C 20 )-aryl-(C1-C 12 )-alkyl,-(C6-C 20 )-aryl-O-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl, -(C3-C 20 )-heteroaryl-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl-O-(C1-C 12-(C1-C) is substituted with one or more substituents selected from alkyl, -COOH, -OH, -SH, -SO3H, -CN, -NH2 or halogens. 30 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -(C6-C 20 )-aryl or-(C3-C 20 Selected from )-heteroaryl groups; X, Y, and Z are independently -O, -NH, -S, -N-(C1-C 30 )-alkyl or -(C1-C 30 ) - Selected from the arrows; [ka] is -CH((C1-C 30 It is -alkyl))-, -(C=O)-, -CH(OH), -SO2-, -SO- or -CH(SOCH3)-; and R4 is -(C1-C 12 )-alkyl,-(C3-C 12 )-Cycloalkyl, -(C3-C 12 )-heterocycloalkyl, -O-(C1-C 12 )-alkyl,-O-(C1-C 12 )-alkyl-(C6-C 20 )-aryl, -O-(C3-C 12 )-Cycloalkyl, -S-(C1-C 12 )-alkyl,-S-(C3-C 12 )-Cycloalkyl, -COO-(C1-C 12 )-alkyl,-COO-(C3-C 12 )-Cycloalkyl, -CONH-(C1-C 12 )-alkyl,-CONH-(C3-C 12 )-Cycloalkyl, -CO-(C1-C 12 )-alkyl,-CO-(C3-C 12 )-Cycloalkyl, -N-[(C1-C 12 )-alkyl]2,-(C6-C 20 )-aryl,-(C6-C 20)-aryl-(C1-C 12 )-alkyl,-(C6-C 20 )-aryl-O-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl, -(C3-C 20 )-heteroaryl-(C1-C 12 )-alkyl,-(C3-C 20 )-heteroaryl-O-(C1-C 12 Selected from )-alkyl, -COOH, -OH, -SH, -SO3H, -CN, -NH2, or halogen.

[0147] Further embodiments include formula IV: [ka] The present invention provides compounds thereof or pharmaceutically acceptable enantiomers, salts, or solvates thereof.

[0148] The compound of formula IV, also known as mJJ64A, can be used in its enantiomers, polymorphs, pharmaceutically acceptable salts, and derivatives to promote or increase the induction of Akt3 physiological activity in immune cells.

[0149] In one embodiment, the Atk3 activator is a derivative of formula I, II, III, or IV. The terms “derivative” or “derivativeization” as used herein include one or more chemical modifications of formula I, II, III, or IV or its enantiomers, polymorphs, or pharmaceutically acceptable salts. That is, a “derivative” can be a functional equivalent of formula I, II, III, or IV that can induce improved pharmacological functional activity and / or behavioral response in a given subject. Examples of such chemical modifications include the substitution of hydrogen with a halo, alkyl, acyl, or amino group.

[0150] Chemical modifications of formulas I, II, III, or IV or their enantiomers, polymorphs, or pharmaceutically acceptable salts may enhance or reduce hydrogen bonding interactions, charge bonding interactions, hydrophobic bonding interactions, van der Waals bonding interactions, or dipole-dipole bonding interactions between the compound and its target.

[0151] In one embodiment, a compound of formula I, formula II, formula III, or formula IV may serve as a model (e.g., a template) for the development of other derivative compounds that are equivalents of the compound and can induce improvements in pharmacological functional activity and / or effects and / or behavioral responses in a given subject.

[0152] Compounds of formulas I, II, III, or IV may be racemic compounds and / or optically active isomers thereof. In this regard, some of the compounds have chiral carbon atoms and therefore may exist as racemic mixtures or individual optical isomers (e.g., enantiomers). Compounds containing a chiral center described herein include all possible stereoisomers of the compound, including compositions containing a racemic mixture of two enantiomers and compositions containing each individual enantiomer substantially without other enantiomers. Thus, for example, intended herein are compositions containing an S enantiomer of a compound substantially without an R enantiomer or an R enantiomer substantially without an S enantiomer. If the compound described contains more than one chiral center, the scope of the present invention also includes compositions containing mixtures of multiple diastereomers in various proportions and compositions containing one or more diastereomers substantially without one or more other diastereomers. "Substantially absent" means that the composition contains the less abundant enantiomer or diastereomer in amounts of approximately 25%, 15%, 10%, 8%, 5%, less than 3%, or less than approximately 1%.

[0153] B. Akt3 inhibitor compounds Compositions and methods for selectively inhibiting Akt3 are disclosed herein. Examples of Akt3 inhibitors are described in U.S. Patent Publications US2017 / 0202956 and 2017 / 0202829 (which are incorporated herein by reference in their entirety), and are listed below.

[0154] 4-[(6-nitroquinoline-4-yl)amino]-N-[4-(pyridine-4-ylamino)phenyl]benzamide was found to selectively inhibit Akt3 activity. 4-[(6-nitroquinoline-4-yl)amino]-N-[4-(pyridine-4-ylamino)phenyl]benzamide has CAS No. 50440-30-7 and the following chemical structure: [ka]

[0155] Other examples of compounds that selectively inhibit Akt3 include the following and their enantiomers, polymorphs, pharmaceutically acceptable salts and derivatives: [ka] [ka]

[0156] In one embodiment, an Akt3 inhibitor is a derivative of any of the disclosed compounds. The terms “derivative” or “derivativeization” as used herein include one or more chemical modifications of any of the disclosed compounds, their enantiomers, polymorphs, or pharmaceutically acceptable salts. That is, a “derivative” may be a functional equivalent of any of the disclosed compounds that can induce improved pharmacological functional activity and / or behavioral response in a given subject. Examples of such chemical modifications include the substitution of hydrogen with a halo, alkyl, acyl, or amino group.

[0157] Chemical modifications of any of the disclosed compounds, or their enantiomers, polymorphs, or pharmaceutically acceptable salts, may enhance or reduce hydrogen bonding interactions, charge bonding interactions, hydrophobic bonding interactions, van der Waals bonding interactions, or dipole bonding interactions between the compound and its target.

[0158] In one embodiment, any of the compounds disclosed may serve as a model (e.g., a template) for the development of other derivative compounds that are equivalents of the compound and can induce improvements in pharmacological functional activity and / or effects and / or behavioral responses in a given subject.

[0159] The disclosed compounds may be racemic compounds and / or optically active isomers. In this regard, some of the compounds have chiral carbon atoms and therefore may exist as racemic mixtures or as individual optical isomers (e.g., enantiomers). Compounds described herein that contain a chiral center include all possible stereoisomers of the compound, including compositions containing a racemic mixture of two enantiomers and compositions containing each individual enantiomer substantially without other enantiomers. Thus, for example, intended herein are compositions containing an S enantiomer of a compound substantially without an R enantiomer or an R enantiomer substantially without an S enantiomer. If the disclosed compound contains more than one chiral center, the scope of the invention also includes compositions containing mixtures of multiple diastereomers in various proportions and compositions containing one or more diastereomers substantially without one or more other diastereomers. "Substantially absent" means that the composition contains the less abundant enantiomer or diastereomer in amounts of approximately 25%, 15%, 10%, 8%, 5%, less than 3%, or less than approximately 1%.

[0160] The disclosed compounds selectively modulate Akt3 compared to Akt1 and Akt2. In one embodiment, none of the disclosed compounds modulate Akt1 and Akt2 to a statistically significant degree. In another embodiment, the modification of Akt3 by the disclosed compounds is about 5, 10, 15, 50, 100, 1000, or 5000 times greater than the modification of Akt1 and / or Akt2.

[0161] C. Immunomodulator or binding site Immunomodulatory agents or binding moieties comprising AKT3 agonists and antagonists are provided. AKT3 agonists typically induce, promote, or enhance AKT3-mediated signaling. AKT3 antagonists typically inhibit, reduce, or block AKT3-mediated signaling. The disclosed compositions and methods may be used to modulate AKT3 and / or counterreceptor signaling in immune cells, including, but not limited to, myeloid cells, T cells, NK cells, or combinations thereof, including, for example, monocytes, Tregs, tumor-associated macrophages (TAMs), myeloid-derived suppressor cells (MDSCs), T cells, Th2 cells, antigen-presenting cells (e.g., monocytes, macrophages, or dendritic cells). In some embodiments, the compositions can specifically target one or more cell types. In some embodiments, the disclosed compositions can be used on tumor cells.

[0162] In some embodiments, an anti-AKT3 agonist induces, promotes, or enhances AKT3-mediated signaling via a known ligand or an unknown counterreceptor, or via an AKT3-binding interaction with said known or unknown counterreceptor. For example, in some embodiments, an AKT3 agonist binds to induce, promote, induce or induce a conformational change or otherwise facilitate AKT3-mediated signaling.

[0163] In one embodiment, an anti-AKT3 antagonist inhibits, reduces, blocks, or otherwise interferes with signaling by blocking the AKT3 binding interaction with a known or unknown counterreceptor via the known or unknown counterreceptor. For example, in one embodiment, an AKT3 antagonist binds to inhibit, block, induces conformational changes, or otherwise interferes with AKT3-mediated signaling.

[0164] 1. Antibodies In one embodiment, the immunomodulator or binding portion is an antibody. A suitable antibody can be prepared by those skilled in the art. The nucleic acid and polypeptide sequences of AKT3 are known in the art, and examples of sequences are provided above. The sequences can be used by those skilled in the art to produce antibodies or antigen-binding fragments specific to AKT3, as will be further detailed below. The antibody or antigen-binding fragment can therefore be an agonist or antagonist of AKT3-mediated signaling.

[0165] The activity of AKT3-specific antibodies or their antigen-binding fragments can be determined using functional assays known in the art, including the assays described below. Typically, the assays involve determining whether the antibody or its antigen-binding fragment increases (i.e., acts as an agonist) or decreases (i.e., acts as an antagonist) AKT3-mediated signaling.

[0166] In one embodiment, the disclosed antibody and antigen-binding fragments bind immunospecifically to human or mouse AKT3. In another embodiment, the antibody binds to the extracellular domain of human or mouse AKT3.

[0167] To prepare antibodies or antigen-binding fragments that specifically bind to AKT3, purified proteins, polypeptides, fragments, fusions, or polypeptides expressed from the AKT3 epitope or its nucleic acid sequence may be used. The antibody or antigen-binding fragment may be prepared using any suitable method known in the art, as further detailed below.

[0168] a. Human and humanized antibodies In one embodiment, the antibody is a humanized antibody. Many non-human antibodies (e.g., those derived from mouse, rat, or rabbit) are inherently antigenic in humans and therefore can produce an unwanted immune response when administered to humans. Therefore, the use of human or humanized antibodies in the method helps reduce the likelihood that an antibody administered to a human may induce an unwanted immune response.

[0169] Transgenic animals (e.g., mice) that can produce a complete repertoire of human antibodies in the absence of endogenous immunoglobulin production through immunization can be used. For example, homozygous deletion of the antibody heavy chain joining region (J(H)) gene in chimeric and germline mutant mice results in complete inhibition of endogenous antibody production. Transmission of a human germline immunoglobulin gene array in such germline mutant mice leads to the production of human antibodies upon antigen loading.

[0170] If desired, antibodies are produced in other species and “humanized” for administration to humans. Humanized non-human (e.g., mouse) antibodies are chimeric immunoglobulins, immunoglobulin chains, or fragments (e.g., Fv, Fab, Fab', F(ab')2, or other antigen-binding subsequences of the antibody) containing the minimal sequence of the non-human immunoglobulin. A humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the complementarity-determining region (CDR) of the recipient antibody are replaced with residues from the CDR of a non-human species (donor antibody), such as mouse, rat, or rabbit, possessing the desired specificity, affinity, and capability. In some examples, Fv framework residues of the human immunoglobulin are replaced with corresponding non-human residues. Humanized antibodies may also contain residues not found in either the recipient antibody or the transferred CDR or framework sequence. Generally, a humanized antibody contains substantially all of the variable domains, at least one and typically two, in which all or substantially all of the CDR region corresponds to that of a non-human immunoglobulin and all or substantially all of the FR region corresponds to the human immunoglobulin consensus sequence. The humanized antibody also optionally and typically contains at least a portion of the immunoglobulin constant region (Fc) of a human immunoglobulin.

[0171] Methods for humanizing non-human antibodies are well known in this field. Generally, humanized antibodies contain one or more amino acid residues introduced from a non-human source. These non-human amino acid residues are often referred to as “import” residues and are typically derived from “import” variable domains. Antibody humanization techniques generally involve the use of recombinant DNA technology to manipulate the DNA sequence encoding one or more polypeptide chains of an antibody molecule. Humanization can be carried out essentially by the substitution of rodent CDRs or CDR sequences with corresponding sequences in human antibodies. Thus, a non-human antibody (or fragment thereof) in humanized form is a chimeric antibody or fragment in which substantially less than the intact human variable domain is substituted with corresponding sequences from a non-human species. In practice, a humanized antibody is typically a human antibody in which some CDR residues and possibly some FR residues are substituted with residues from similar positions in the rodent antibody.

[0172] The selection of both light and heavy human variable domains for the production of humanized antibodies is crucial for reducing antigenicity. A "best fit" method screens the variable domain sequences of rodent antibodies against a complete library of known human variable domain sequences. The human sequence most closely resembling that of a rodent is then accepted as the human framework (FR) for the humanized antibody. Another method uses a specific framework derived from the consensus sequences of all human antibodies for a particular subgroup of the light or heavy chain. The same framework may be used for several different humanized antibodies.

[0173] It is even more important that antibodies are humanized while retaining high affinity for antigens and other advantageous biological properties. To achieve this objective, humanized antibodies can be prepared by analytical processes of parental sequences and various conceptual humanized products using three-dimensional models of parental and humanized sequences. Three-dimensional immunoglobulin models are generally available and familiar to those skilled in the art. Computer programs are available that describe and present the likely three-dimensional structure of selected candidate immunoglobulin sequences. By investigating these presentations, it becomes possible to analyze the likely roles of residues in the function of candidate immunoglobulin sequences, i.e., the residues that affect the ability of the candidate immunoglobulin to bind to the antigen. In this method, FR residues are selected and combined from consensus and translocation sequences so that desired antibody characteristics, such as increased affinity for the target antigen, are achieved. In general, CDR residues are directly and most substantially involved in the effect on antigen binding.

[0174] The antibody may be conjugated to a substrate, labeled with a detectable portion, or both conjugated and labeled. Detectable portions considered in this composition include fluorescence, enzymes, and radiomarkers.

[0175] b. Single chain antibody In one embodiment, the antibody is a single-chain antibody. Methods for producing single-chain antibodies are well known to those skilled in the art. Single-chain antibodies are reconstructed into a single molecule by fusing the variable domains of the heavy and light chains using a short peptide linker. Single-chain antibody variable fragments (scFvs), in which the C-terminus of one variable domain is linked to the N-terminus of the other variable domain via a 15-25 amino acid peptide or linker, have been developed without significantly disrupting antigen binding or binding specificity. The linker is selected to allow the heavy and light chains to be bound in appropriate stereoorientation. These Fvs lack the constant region (Fc) present in the heavy and light chains of native antibodies.

[0176] c. Monovalent antibodies In one embodiment, the antibody is a monovalent antibody. In vitro methods are also suitable for the preparation of monovalent antibodies. Digestion of the antibody to produce its fragments, particularly Fab fragments, can be achieved using routine techniques known in the art. For example, digestion can be carried out using papain. Papain digestion of the antibody produces two identical antigen-binding fragments, typically referred to as Fab fragments, each consisting of a single antigen-binding site and a residual Fc fragment. Pepsin treatment yields a fragment, referred to as an F(ab')2 fragment, which has two antigen-combination sites and is still antigen-crosslinkable.

[0177] Fab fragments produced by antibody digestion also contain a constant domain of the light chain and a primary constant domain of the heavy chain. Fab' fragments differ from Fab fragments by the addition of several residues at the carboxyl terminus of the heavy chain domain, including one or more cysteines from the antibody hinge region. F(ab')2 fragments are bivalent fragments containing two Fab' fragments linked by disulfide crosslinks in the hinge region. Fab'-SH here refers to Fab' fragments in which the cysteine ​​residues of the constant domain contain free thiol groups. Antibody fragments were originally produced from pairs of Fab' fragments with hinge cysteines in between. Other chemical couplings of antibody fragments are also known.

[0178] d. Hybrid antibodies In one embodiment, the antibody is a hybrid antibody. In a hybrid antibody, one heavy-chain and light-chain pair is homologous to those found in an antibody induced against a certain epitope, while the other heavy-chain and light-chain pair is homologous to those found in an antibody induced against another epitope. This results in a multifunctional titration property, i.e., the ability to bind to at least two different epitopes simultaneously. Such hybrids can be formed by fusion or recombination techniques of hybridomas that produce each component antibody. Such hybrids can, of course, also be formed using chimeric chains.

[0179] e. Conjugate or fusion of antibody fragments In one embodiment, the antibody is a conjugate or fusion of antibody fragments. The targeting function of the antibody can be used therapeutically by coupling the antibody or its fragment with a therapeutic agent. Such coupling of an antibody or fragment (e.g., at least a portion of the constant region (Fc) of an immunoglobulin) with a therapeutic agent can be achieved by the production of an immunoconjugate or fusion protein comprising the antibody or antibody fragment and the therapeutic agent.

[0180] Such coupling of an antibody or fragment with a therapeutic agent can be achieved by the production of an immunoconjugate containing the antibody or antibody fragment and the therapeutic agent, or by the production of a fusion protein, or by the linkage of the antibody or fragment to a nucleic acid such as siRNA.

[0181] In one embodiment, the antibody is modified to alter its half-life. In one embodiment, it is desirable to extend the half-life of the antibody so that it remains in circulation or at the treatment site for a longer period of time. For example, it may be desirable to maintain the antibody titer for a longer period of time at the circulation or treatment site. The antibody is modified, for example, by Xtend TMThe antibody half-life can be extended using antibody half-life extension technology (Xencor, Monrovia, CA). In other embodiments, the half-life of an anti-DNA antibody is shortened to reduce the potential for side effects. The disclosed conjugates can be used to modify certain biological responses. The drug moiety is not construed to be limited to classical chemotherapeutic agents. For example, the drug moiety may be a protein or polypeptide having the desired biological activity. Such proteins are toxins such as abrin, lysine A, Pseudomonas exotoxin, or diphtheria toxin.

[0182] 2. Proteins and polypeptides a. Protein and polypeptide compositions The immunomodulator or binding agent may be an AKT3 protein, polypeptide, or fusion protein. For example, the immunomodulator or binding site may be an isolated or recombinant AKT3 protein or polypeptide, or a functional fragment, variant, or fusion protein thereof.

[0183] AKT3 protein or polypeptide or its functional fragment, variant, or fusion protein can be an agonist or an antagonist. For example, in one embodiment, an AKT3 antagonist is an AKT3 polypeptide or its fragment or fusion protein that binds to an AKT3 ligand. The polypeptide is a soluble fragment, for example, the extracellular domain of AKT3 or its functional fragment or fusion protein. In one embodiment, a soluble ligand of AKT3 may serve as an antagonist that reduces AKT3-mediated signaling.

[0184] The activity of AKT3 proteins or polypeptides or any fragments, variants, or fusion proteins thereof can be determined using functional assays known in the art, including the assays described below. Typically, the assay involves determining whether a protein, polypeptide or its fragments, variants, or fusion proteins increase (i.e., agonists) or decrease (i.e., antagonists) AKT3 receptor-mediated signaling. In some embodiments, the assay involves determining whether a protein, polypeptide or its fragments, variants, or fusion proteins increase (i.e., agonists) or decrease (i.e., antagonists) AKT3-related immune responses. Typically, the assay involves determining whether a protein, polypeptide or its fragments, variants, or fusion proteins increase (i.e., agonists) or decrease (i.e., antagonists) AKT3-mediated signaling. In some embodiments, the assay involves determining whether a protein, polypeptide or its fragments, variants, or fusion proteins decrease (i.e., agonists) or increase (i.e., antagonists) AKT3-regulated immune responses. In one embodiment, the assay includes determining whether a protein, polypeptide or its fragment, variant, or fusion protein reduces the self-renewal capacity of AML and ALL stem cells, or increases apoptosis and differentiation (i.e., antagonists) of acute myeloid leukemia (AML) cells and acute lymphoblastic leukemia (ALL) cells.

[0185] Nucleic acid and polypeptide sequences of AKT3 are known in the art, and examples of protein and peptide sequences are provided above. These sequences can be used by those skilled in the art to produce any AKT3 protein or polypeptide or any fragment, variant, or fusion protein of AKT3, as further detailed below. Generally, AKT3 proteins, polypeptides, their fragments, variants, and fusions are expressed from nucleic acids containing a signal sequence. The signal sequence is generally cleaved from immature polypeptides to produce mature polypeptides lacking the signal sequence. The signal sequence can be replaced with the signal sequence of other polypeptides using standard molecular biology techniques to affect the expression level, secretion, solubility, or other properties of AKT3 polypeptide proteins, whether they have a signal sequence or not. In some cases, mature proteins known or described in the art, i.e., protein sequences without a signal sequence, are putatively mature proteins. During normal cellular expression, the signal sequence may be removed by cellular peptidases to become a mature protein. The sequences of mature proteins can be determined or confirmed using methods known in the art.

[0186] i. Fragment The AKT3 fragments used herein refer to any subset of polypeptides that are at least one amino acid shorter than the full-length protein. Useful fragments include those that retain the ability to bind to the native ligand or ligand. Polypeptides that are fragments of any full-length AKT3 typically have an ability to bind to the native ligand at least approximately 20 percent, 30 percent, 40 percent, 50 percent, 60 percent, 70 percent, 80 percent, 90 percent, 95 percent, 98 percent, 99 percent, 100 percent, or more than 100 percent compared to the full-length protein.

[0187] AKT3 fragments include cell-free fragments. Cell-free polypeptides can be fragments of a full-length, transmembrane polypeptide that can be detached, secreted, or otherwise extracted from producing cells. Cell-free fragments of polypeptides may contain some or all of the extracellular domain of the polypeptide and lack some or all of the intracellular and / or transmembrane domains of the full-length protein. In one embodiment, the polypeptide fragment contains the entire extracellular domain of the full-length protein. In other embodiments, the cell-free fragment of a polypeptide contains a fragment of the extracellular domain that retains the biological activity of the full-length protein. The extracellular domain may contain 1, 2, 3, 4, or 5 consecutive amino acids of the transmembrane domain and / or 1, 2, 3, 4, or 5 consecutive amino acids of the signal sequence. Alternatively, the extracellular domain may have 1, 2, 3, 4, 5, or more amino acids removed from the C-terminus, N-terminus, or both. In one embodiment, the extracellular domain is the sole functional domain of the fragment (e.g., the ligand-binding domain).

[0188] ii. Variants Variants and fragments of AKT3 are also provided. In some embodiments, the variants are at least about 50 percent, 60 percent, 70 percent, 80 percent, 85 percent, 90 percent, 95 percent, 96 percent, 97 percent, 98 percent, or 99 percent identical to either SEQ ID NO: 1 or 2. Useful variants include those that increase the biological activity or the half-life or stability of the protein as shown by any of the assays described herein. AKT3 proteins and polypeptides and their fragments, variants, and fusion proteins can be manipulated to increase biological activity. For example, in some embodiments, the AKT3 polypeptide, protein or its fragments, variants, or fusions are modified by at least one amino acid substitution, deletion, or insertion to increase function.

[0189] Finally, variant polypeptides can be manipulated to have an extended half-life compared to the wild type. These variants are generally modified to be resistant to enzymatic degradation. Examples of modifications include modified amino acid residues and modified peptide bonds to be resistant to enzymatic degradation. Various modifications to achieve this are known in this field. Variants can be modified to modulate the effect of receptor affinity on the half-life of proteins, polypeptides, fragments, or fusions at serum and endosomal pH.

[0190] iii. Fusion protein The fusion polypeptide has a first fusion partner comprising all or part of a human or mouse AKT3 polypeptide, fused directly to the second polypeptide or via a linker peptide sequence fused to the second polypeptide. In one embodiment, the ECD or a fragment of human or mouse AKT3 is fused to the second polypeptide. The fusion protein optionally includes domains that function to dimerize or polymerize two or more fusion proteins. The peptide / polypeptide linker domain may be a separate domain or separately contained in one of the other domains of the fusion protein (the first polypeptide or the second polypeptide). Similarly, the domain that functions to dimerize or polymerize the fusion protein may be a separate domain or separately contained in one of the other domains of the fusion protein (the first polypeptide or the second polypeptide). In one embodiment, the dimerization / polymerization domain and the peptide / polypeptide linker domain are the same.

[0191] The fusion protein disclosed herein is given by formula A: N-P1-P2-P3-C Here, "N" represents the N-terminus of the fusion protein, and "C" represents the C-terminus of the fusion protein. In one embodiment, "P1" is the polypeptide or protein of AKT3 or a fragment or variant thereof, "P2" is an arbitrary peptide / polypeptide linker domain, and "P3" is a secondary polypeptide. Alternatively, P3 may be the polypeptide or protein of AKT3 or a fragment or variant thereof, and P1 may be a secondary polypeptide. In one embodiment, the AKT3 polypeptide is an extracellular domain.

[0192] Dimerization or polymerization can occur between two or more fusion proteins via dimerizing or polymerizing domains. Alternatively, dimerization or polymerization of fusion proteins can occur through chemical crosslinking. The resulting dimers or polymers may be homodimers / homopolymers or heterodimers / heteropolymers.

[0193] In one embodiment, the fusion protein comprises the extracellular domain or fragment or variant of AKT3 fused to the Ig Fc region, as previously described (Chapoval, et al., Methods Mol. Med., 45:247-255 (2000); incorporated herein by reference as a whole). The recombinant Ig fusion protein may be produced by fusing the coding region of the extracellular domain or fragment or variant to the Fc region of human IgG1, IgG2, IgG3, or IgG4, or mouse IgG2a, or other suitable Ig domain.

[0194] iv. Polypeptide modification Polypeptides and fusion proteins can be modified with chemical moieties that may be present in polypeptides in the normal cellular environment, such as phosphorylation, methylation, amidation, sulfation, acylation, glucosylation, smolation, and ubiquitination. Fusion proteins can also be modified with labels that provide detectable signals, directly or indirectly, including but not limited to radioisotopes and fluorescent compounds.

[0195] Polypeptides and fusion proteins can also be modified by chemical moieties not typically added to polypeptides in the cellular environment. For example, the disclosed fusion proteins may also be modified by covalent bonding of polymer chains, including but not limited to polyethylene glycol polymer (PEG) chains (i.e., pegylation). Conjugation of macromolecules to PEG has recently emerged as an effective strategy for modifying the pharmacokinetic (PK) profiles of various drugs and thereby improving their therapeutic efficacy. PEG conjugation increases the retention of drugs in circulation by protecting against enzymatic digestion, slowing renal filtration, and reducing neutralizing antibody production. Furthermore, PEG conjugates can be used to enable the multimerization of fusion proteins.

[0196] Modifications can be introduced into a molecule by the reaction of a target amino acid residue of the polypeptide with an organic derivatizer that can react with a selected side chain or terminal residue. Another modification is protein cyclization.

[0197] Examples of chemical derivatives of polypeptides include ricinyl and amino-terminal residues derivatized with succinic anhydride or other carboxylic acid anhydrides. Derivatization with cyclic carboxylic acid anhydrides has the effect of reversing the charge of the ricinyl residue. Other suitable agents for amino-containing residue derivatization include imide esters such as methyl picolinimide; pyridoxal phosphate; pyridoxal; chlorobolohydride; trinitrobenzenesulfonic acid; O-methylisourea; 2,4-pentanedione; and transaminase-catalyzed reactions with glyoxylic acid. The carboxyl side chain group, aspartyl, or glutamyl can be selectively modified by reaction with carbodiimides (RN=C=N-R') such as 1-cyclohexyl-3-(2-morpholinyl-(4-ethyl)carbodiimide or 1-ethyl-3-(4-azonia-4,4-dimethylpentyl)carbodiimide. Furthermore, aspartyl and glutamyl residues can be converted to asparaginyl and glutamyl residues by reaction with ammonia. The fusion protein may also contain one or more D-amino acids that are substitutes for one or more L-amino acids.

[0198] v. Modified bond properties The binding properties of proteins, polypeptides, their fragments, variants, and fusions are related to the dose and dose regimen to be administered. In some embodiments, the disclosed proteins, polypeptides, their fragments, variants, and fusions have binding properties to AKT3 or AKT3 ligands such that the occupancy of the binding site (e.g., on the ligand) is long-lasting or high-percentage compared to other receptor molecules to which they bind. In other embodiments, the disclosed proteins, polypeptides, their fragments, variants, and fusions have reduced binding affinity to AKT3 compared to the wild-type protein.

[0199] In one embodiment, the protein, polypeptide, its fragments, variants, and fusions have a relatively high affinity for AKT3 and therefore have a relatively slow dissociation rate. In another embodiment, the protein, polypeptide, its fragments, variants, and fusions are administered intermittently over a period of several days, weeks, or months to attenuate the immune response, which may help to modify the immune response without completely initiating or stopping it, allowing for recovery before the next administration and thus avoiding long-term side effects.

[0200] 3. Isolated nucleic acid molecules Isolated nucleic acid sequences encoding the AKT3 protein, polypeptide, its fragments, variants, and fusions are disclosed herein. As used herein, “isolated nucleic acid” refers to nucleic acid separated from other nucleic acid molecules present in the mammalian genome, including nucleic acids that are normally adjacent to one or both sides of the nucleic acids in the mammalian genome. As used herein in relation to nucleic acids, “isolation” includes any combination of nucleic acid sequences that do not exist in nature, because such non-natural sequences are not inherently found and do not have immediately contiguous sequences in the naturally occurring genome.

[0201] An isolated nucleic acid can be a DNA molecule, for example, insofar as one nucleic acid sequence immediately adjacent to a DNA molecule in a naturally occurring genome is removed or deleted. Therefore, isolated nucleic acids include, but are not limited to, DNA molecules existing as separate molecules (e.g., chemically synthesized nucleic acids or cDNA or genomic DNA fragments produced by PCR or restriction endonuclease treatment), as well as vectors, self-replicating plasmids, viruses (e.g., retroviruses, lentiviruses, adenoviruses, or herpesviruses) or recombinant DNA incorporated into the genomic DNA of prokaryotes or eukaryotes. Furthermore, isolated nucleic acids include engineered nucleic acids, such as recombinant DNA molecules that are part of a hybrid or fusion nucleic acid. Nucleic acids present in hundreds of millions or millions of nucleic acids, such as in a cDNA library or genomic library, or a gel slice containing a restriction digest of genomic DNA, are not considered isolated nucleic acids.

[0202] Nucleic acids encoding proteins, polypeptides, their fragments, variants, and fusions can be optimized for expression in a selected expression host. Codons can be substituted with other codons encoding the same amino acid, taking into account codon usage frequency between the mammal from which the nucleic acid sequence originates and the expression host. In this way, nucleic acids can be synthesized using expression host-preferred codons.

[0203] Nucleic acids may be sense or antisense oriented or may be complementary to a control sequence encoding a polypeptide or protein of AKT3. Nucleic acids may be DNA, RNA, or nucleic acid analogs. Nucleic acid analogs may be modified with base moieties, sugar moieties, or phosphate backbones. Such modifications may improve, for example, the stability, hybridization, or solubility of the nucleic acid. Modifications of base moieties may include deoxyuridine for deoxythymidine and 5-methyl-2'-deoxycytidine or 5-bromo-2'-deoxycytidine for deoxycytidine. Modifications of sugar moieties may include modifications of the 2'-hydroxyl group of ribose sugars that form 2'-O-methyl or 2'-O-allyl sugars. Deoxyribose phosphate backbones can be modified to produce morpholino nucleic acids in which each base moiety is 6-membered and linked to a morpholino ring, or peptide nucleic acids in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and 4 bases are retained. For example, see Summerton and Weller (1997) Antisense Nucleic Acid Drug Dev. 7:187-195; and Hyrup et al. (1996) Bioorg. Med. Chem. 4:5-23 (each incorporated herein by reference as a whole). Furthermore, the deoxyphosphate backbone can be replaced with, for example, a phosphorothioate or phosphorodithioate backbone, a phosphoramidite, or an alkylphosphotryester backbone.

[0204] Nucleic acids encoding polypeptides can be administered to targets that require them. Nucleic acid delivery includes the introduction of “foreign” nucleic acids into cells and, ultimately, into living animals. Compositions and methods for delivering nucleic acids to targets are known in the art (see Understanding Gene Therapy, Lemoine, NR, ed., BIOS Scientific Publishers, Oxford, 2008; incorporated herein by reference in whole).

[0205] 4. Vector and host cell Vectors encoding proteins, polypeptides, their fragments, variants, and fusions are also provided. Such nucleic acids may be inserted into vectors for expression in cells. As used herein, “vector” refers to a replicon such as a plasmid, phage, virus, or cosmid into which other DNA segments may be inserted, resulting in replication of the inserted segment. A vector may be an expression vector. An “expression vector” is a vector containing one or more expression regulatory sequences, where “expression regulatory sequences” are DNA sequences that regulate and control the transcription and / or translation of other DNA sequences.

[0206] Nucleic acids in a vector can be operably ligated with one or more regulatory sequences. "Operablely ligated" here means that the regulatory sequences are incorporated into the gene construct so as to efficiently control the expression of the target coding sequence. Examples of regulatory sequences include promoters, enhancers, and transcription stop regions. A promoter is a regulatory sequence consisting of a region of the DNA molecule, typically within 100 nucleotides upstream of the transcription start point (generally near the RNA polymerase II start site). To place the coding sequence under the control of a promoter, the translation start region of the polypeptide's translation reading frame must be positioned 1 to approximately 50 nucleotides downstream of the promoter. Enhancers provide expression specificity in terms of time, place, and level. Unlike promoters, enhancers can function at various distances from the transcription site. Enhancers can also be located downstream of the transcription start site. A coding sequence is "operably ligated" and "under control" with a regulatory sequence in a cell if RNA polymerase can transcribe the coding sequence into mRNA, which can then be translated into the protein encoded by the coding sequence.

[0207] Suitable expression vectors include, but are not limited to, plasmids and viral vectors derived from, for example, bacteriophages, baculoviruses, tobacco mosaic viruses, herpesviruses, cytomegaloviruses, retroviruses, vaccinia viruses, adenoviruses, and adeno-associated viruses. Numerous vectors and expression systems are commercially available from vendors such as Novagen (Madison, WI), Clontech (Palo Alto, CA), Stratagene (La Jolla, CA), and Invitrogen Life Technologies (Carlsbad, CA).

[0208] Expression vectors may contain tag sequences. Tag sequences are typically expressed as fusions with the encoding polypeptide. Such tags can be inserted anywhere in the polypeptide, including at the carboxyl or amino terminus. Examples of useful tags include green fluorescent protein (GFP), glutathione-transferase (GST), polyhistidine, c-myc, hemagglutinin, and Flag. TM The disclosed polypeptides include, but are not limited to, tags (Kodak, New Haven, CT), maltose E-binding proteins, and protein A. In one embodiment, a nucleic acid molecule encoding one of the disclosed polypeptides is present in a vector containing nucleic acids encoding one or more domains of the constant region of the Ig heavy chain, having amino acid sequences corresponding to the hinge, CH2 and CH3 regions of the human immunoglobulin Cγ1 chain.

[0209] Vectors containing nucleic acids to be expressed can be transmitted to host cells. The term “host cell” is intended to include prokaryotic and eukaryotic cells to which recombinant expression vectors can be transmitted. As used herein, “transform” and “transfect” include the introduction of nucleic acid molecules (e.g., vectors) into cells by one of several techniques. While not limited to specific techniques, several of these techniques are well established in this field. Prokaryotic cells can be transformed with nucleic acids, for example, by electroporation or calcium chloride-mediated transformation. Nucleic acids can be transfected into mammalian cells by techniques including, for example, calcium phosphate coprecipitation, DEAE-dextran-mediated transfection, lipofection, electroporation, or microinjection. Host cells (e.g., prokaryotic or eukaryotic cells such as CHO cells) can be used to produce, for example, the proteins, polypeptides, their fragments, variants, and fusions described herein.

[0210] The vectors described may be used to express proteins, polypeptides, their fragments, variants, and fusions in cells. Examples of vectors include, but are not limited to, adenovirus vectors. One method involves nucleic acid transfer to primary cells in culture, followed by ex vivo autotransplantation of the transformed cells into a host, either systemically or to a specific organ or tissue. Ex vivo methods include, for example, collecting cells from a subject, culturing the cells, transducing them using an expression vector, and maintaining the cells under conditions suitable for the expression of the encoding polypeptide. These methods are known in the field of molecular biology. The transduction step may be achieved by any of the standard methods used in ex vivo gene therapy, including, for example, calcium phosphate, lipofection, electroporation, viral infection, and microparticle gun gene transfer. Alternatively, liposomes or polymer microparticles may be used. Cells that have been successfully transduced may then be selected for, for example, the expression of a coding sequence or a drug resistance gene. The cells may then be lethally irradiated (optionally) and injected or transplanted into a subject. In one embodiment, an expression vector containing nucleic acids encoding a fusion protein is transfected into cells to be administered to a target that requires it.

[0211] In vivo nucleic acid therapy can be achieved by directly delivering functionally active DNA to mammalian somatic tissues or organs in vivo. For example, nucleic acids encoding the polypeptides disclosed herein can be directly administered to lymphoid tissues. Alternatively, lymphoid tissue-specific targeting can be achieved using lymphoid tissue-specific transcription factors (TREs), such as B lymphocyte, T lymphocyte, or dendritic cell-specific TREs. Lymphoid tissue-specific TREs are known in this field.

[0212] Nucleic acids can also be administered in vivo by viral means. Nucleic acid molecules encoding fusion proteins can be packaged into retroviral vectors using packaging cell lines that produce replication-deficient retroviruses, as is well known in this art. Other viral vectors, including recombinant adenoviruses and vaccinia viruses that can be made non-replicating, can also be used. In addition to naked DNA or RNA or viral vectors, engineered bacteria can be used as vectors.

[0213] Nucleic acids can also be delivered by liposomes, polymer micros and nanoparticles, and other carriers containing polycations such as asialoclycoproteins / polylysine.

[0214] In addition to in vivo viral and carrier-mediated gene transfer, well-known physical means in this field, including plasmid DNA administration and particulate gun-mediated gene transfer, can be used for direct DNA transfer.

[0215] 5. Small molecules Immunomodulators can be small molecules. Small molecule agonists and antagonists AKT3 can be identified using methods known in this field or routine screening methods.

[0216] In one embodiment, the screening assay may include random screening of a large library of test compounds. Alternatively, the assay may be used to focus on a specific class of compounds suspected of modulating AKT3 levels. The assay may include determining AKT3-mediated signaling activity. Other assays may include determining nucleic acid transcription or translation, mRNA levels, mRNA stability, mRNA degradation, transcription rate, and translation rate.

[0217] D. Pharmaceutical Compositions One embodiment provides formulations of the disclosed Akt3 activator or inhibitor and pharmaceutical compositions comprising the same. Generally, the dosage levels of the compounds disclosed herein range from about 0.0001 mg / kg body weight to about 1,000 mg / kg body weight, more preferably 0.001 to 500 mg / kg body weight, and more preferably 0.01 to 50 mg / kg body weight per day when administered to a mammal.

[0218] Pharmaceutical compositions comprising an Akt3 modulator, with or without a delivery medium, are provided. The pharmaceutical compositions can be formulated for administration via non-enteral (intramuscular, intraperitoneal, intravenous (IV), or subcutaneous), enteral, transmucosal (nasal, vaginal, rectal, or sublingual), or transdermal (passive or using ion electrophoresis or electroporation) routes of administration, or using biodegradable inserts, and can be formulated into dosage forms suitable for each route of administration.

[0219] In one embodiment, the composition is administered locally, for example, by direct injection into the site to be treated (e.g., a tumor). In another embodiment, the composition is administered directly by injecting it into a vascular structure on vascular tissue at or near the site to be treated (e.g., near a tumor). Typically, local administration results in a greater increase in localized concentration than that achievable with systemic administration of the composition.

[0220] Non-enteral administration formulation Compounds and their pharmaceutical compositions may be administered by non-enteral injection in aqueous solution. The formulations may be in the form of suspensions or emulsions. Generally, pharmaceutical compositions are provided that contain an effective amount of the activator and optionally include pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants and / or carriers. Such compositions include a diluent of sterile water, various buffer contents (e.g., Tris-HCl, acetic acid, phosphoric acid), buffered saline of pH and ionic strength; and optionally, surfactants and solubilizers (e.g., twine, also known as polysorbate 20 or 80). (登録商標) 20. Twin (登録商標)80) Additives include antioxidants (e.g., ascorbic acid, sodium metabisulfite), preservatives (e.g., thimerosal, benzyl alcohol), and fillers (e.g., lactose, mannitol). Examples of non-aqueous solvents or media include propylene glycol, polyethylene glycol, vegetable oils such as olive oil and corn oil, and injectable organic esters such as gelatin and ethyl oleate. The formulation may be lyophilized and redissolved / resuspended immediately before use. The formulation may be sterilized, for example, by filtration through a bacterial-retaining filter, incorporation of a sterilizing agent into the composition, irradiation of the composition, or heating of the composition.

[0221] 2. Enteral preparations Suitable oral dosage forms include tablets, capsules, solutions, suspensions, syrups, and lozenges. Tablets can be manufactured using compression or casting techniques well known in the art. Gelatin or non-gelatin capsules can be prepared using techniques well known in the art as hard or soft capsule shells capable of encapsulating liquid, solid, and semi-solid filling materials.

[0222] The formulation may be prepared using a pharmaceutically acceptable carrier. Generally as used herein, “carrier” includes, but is not limited to, diluents, preservatives, binders, lubricants, disintegrants, swelling agents, fillers, stabilizers, and combinations thereof.

[0223] The carrier also includes all components of the coating composition, which may contain plasticizers, dyes, colorants, stabilizers, and flow enhancers. Delayed-release dosage forms can be prepared as described in standard reference books. These reference books provide information on carriers, materials, apparatus, and processes for tablets and capsules, as well as for tablets, capsules, and granules in delayed-release dosage forms.

[0224] Examples of suitable coating agents include cellulose polymers such as cellulose acetate phthalate, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and hydroxypropyl methylcellulose acetate succinate; polyvinyl acetate phthalate, acrylic polymers and copolymers, and trade name Eudragit. (登録商標) This includes, but is not limited to, commercially available methacrylic resins, zein, shellac, and polysaccharides under Roth Pharma, Westerstadt, Germany.

[0225] Furthermore, the coating agent may contain conventional carriers such as plasticizers, dyes, colorants, flow promoters, stabilizers, pore-forming agents, and surfactants.

[0226] Any pharmaceutically acceptable additives include, but are not limited to, diluents, binders, lubricants, disintegrants, colorants, stabilizers, and surfactants. Diluents, also called “fillers,” are typically needed to increase the bulk of solid dosage forms so that tablet compression or bead and granule formation is provided. Suitable diluents include, but are not limited to, dicalcium phosphate dihydrate, calcium sulfate, lactose, sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, kaolin, sodium chloride, dried starch, hydrolyzed starch, pregelatinized starch, silicon dioxide, titanium dioxide, aluminum magnesium silicate, and powdered sugars.

[0227] Binders are used to impart tackiness to solid dosage forms, ensuring that tablets, beads, or granules remain intact after formation into dosage forms. Suitable binder materials include, but are not limited to, starch, pregelatinized starch, gelatin, sugars (including sucrose, glucose, dextrose, lactose, and sorbitol), polyethylene glycol, wax, natural and synthetic gums, such as acacia, tragacanth, sodium alginate, and cellulose and bee gums, including hydroxypropyl methylcellulose, hydroxypropylcellulose, and ethylcellulose, and synthetic polymers, such as acrylic acid and methacrylic acid copolymers, methacrylic acid copolymers, methyl methacrylic acid copolymers, aminoalkyl methacrylic acid copolymers, polyacrylic acid / polymethacrylic acid, and polyvinylpyrrolidone.

[0228] Lubricants are used to facilitate the manufacture of tablets. Suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, glycerol behenate, polyethylene glycol, talc, and mineral oil.

[0229] Disintegrants are used to accelerate the breakdown or "decomposition" of the dosage form after administration, and commonly include starch, sodium starch glycolate, sodium carboxymethyl starch, sodium carboxymethylcellulose, hydroxypropylcellulose, pregelatinized starch, clay, cellulose, arginine, gum, or cross-linked PVP (Polyplasdone from GAF Chemical Corp). (登録商標) This includes, but is not limited to, cross-linked polymers such as XL.

[0230] Stabilizers are used, for example, to inhibit or delay drug degradation reactions, including oxidation reactions. Suitable stabilizers include, but are not limited to, antioxidants, butylated hydroxytoluene (BHT); ascorbic acid, its salts and esters; vitamin E, tocopherol and its salts; sulfites such as sodium metabisulfite; cysteine ​​and its derivatives; citric acid; propyl gallate and butylated hydroxyanisole (BHA).

[0231] Oral administration forms such as capsules, tablets, solutions, and suspensions can be formulated for controlled release. For example, one or more compounds and any one or more additional activators can be formulated into nanoparticles, microparticles, or combinations thereof, and encapsulated in soft or hard gelatin or non-gelatin capsules or dispersed in a dispersion medium to form an oral suspension or syrup. The particles may be in the form of the drug and a controlled-release polymer or matrix. Alternatively, the drug particles may be coated with one or more controlled-release coatings before being incorporated into the final administration form.

[0232] In another embodiment, one or more compounds and one or more additional activators are dispersed in a matrix material that gels or emulsifies upon contact with an aqueous medium such as body fluids. In the case of a gel, the matrix expands, encapsulating the activators, which are slowly released over time by diffusion and / or decomposition of the matrix material. Such matrices can be formulated as fillers for tablets or hard and soft capsules.

[0233] In yet another embodiment, one or more compounds and one or more additional activators are formulated into commercially available oral administration forms such as tablets or capsules, and the solid administration forms are coated with one or more controlled-release coatings such as delayed-release coatings or sustained-release coatings. One or more coatings may also include the compound and / or additional activators.

[0234] Continuous release administration Sustained-release formulations are generally prepared as diffusion or permeation systems, as is well known in the art. Diffusion systems typically consist of two types of devices: reservoirs and matrices, which are well known and described in the art. Matrix devices are generally prepared by compressing a drug-gradually soluble polymer carrier into tablet form. The three main types of materials used in the preparation of matrix devices are insoluble plasticizers, hydrophilic polymers, and fatty compounds. Plastic matrices include, but are not limited to, methyl acrylate-methyl methacrylate, polyvinyl chloride, and polyethylene. Hydrophilic polymers include cellulose polymers, e.g., methyl and ethyl cellulose; hydroxyalkyl celluloses, e.g., hydroxypropyl cellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and Carbopol. (登録商標) 934 includes, but is not limited to, polyethylene oxide and mixtures thereof. Fatty compounds include, but are not limited to, various waxes, such as carnauba wax and glyceryl tristearate and waxy substances or mixtures thereof, including hydrogenated castor oil or hydrogenated vegetable oil.

[0235] In one preferred embodiment, the plastic material is a pharmaceutically acceptable acrylic acid polymer, including but not limited to acrylic acid and methacrylic acid copolymers, methyl methacrylate, methyl methacrylate copolymer, ethoxyethyl methacrylate, cyanoethyl methacrylate, aminoalkyl methacrylate copolymer, poly(acrylic acid), poly(methacrylic acid), alkylamine methacrylate copolymer, poly(methyl methacrylate), poly(methacrylic acid) (anhydride), polymethacrylate, polyacrylamide, poly(methacrylic anhydride), and glycidyl methacrylate copolymer.

[0236] In one preferred embodiment, the acrylic acid polymer consists of one or more ammonia methacrylate copolymers. Ammonia methacrylate copolymers are well known in the art and are described in NF XVII as complete polymerization copolymers of acrylic acid and methacrylate esters having a low content of quaternary ammonium groups.

[0237] In one preferred embodiment, the acrylic acid polymer is traded as Eudragit. (登録商標) The acrylic resin lacquer is commercially available from Rohm Pharma, for example. In a more preferred embodiment, the acrylic polymers are each trade name Eudragit (登録商標) RL30D and Oidragit (登録商標) RS30D contains a mixture of two acrylic resin lacquers commercially available from Rohm Pharma. (登録商標) RL30D and Oidragit (登録商標) RS30D is a copolymer of acrylic acid and methacrylic acid ester having a low content of quaternary ammonium groups, and the molar ratio of ammonium groups to the remaining neutral (methacrylic) ester is Eudragit. (登録商標) RL30D 1:20 and Oidragit (登録商標) The ratio is 1:40 in RS30D. The average molecular weight is approximately 150,000. (Eudragit) (登録商標) S-100 and Oidragit (登録商標) L-100 is also preferable. The code names RL (high permeability) and RS (low permeability) refer to the permeability of these drugs. Eudragit (登録商標) RL / RS mixtures are insoluble in water and digestate. However, multiplicative systems formed to contain them are swellable and permeable to aqueous solutions and digestate.

[0238] Oidragit (登録商標) The above polymers, such as RL / RS, can be mixed in any desired ratio to obtain a sustained-release formulation with the desired solubility profile. The desired sustained-release multiparticle system is, for example, 100% Eudragit. (登録商標) RL, 50% Oidragit(登録商標) RL and 50% Oidragit (登録商標) RS and 10% Oudragit (登録商標) RL and 90% Oidragit (登録商標) It can be obtained from RS. Those skilled in the art will know, for example, Eudragit (登録商標) It should be recognized that other acrylic acid polymers, such as L, may also be used.

[0239] Alternatively, sustained-release formulations can be prepared using an infiltration system or by applying a semipermeable coating to the dosage form. In the latter case, the desired drug release profile can be achieved by combining low-permeability and high-permeability coatings in appropriate ratios.

[0240] Devices with the different drug release mechanisms described above can be combined in a final dosage form containing one or more units. Examples of multiple units include, but are not limited to, multilayer tablets and capsules containing tablets, beads, or granules.

[0241] An immediate-release portion can be added to a sustained-release system by applying an immediate-release layer to the top of a sustained-release core using a coating or compression process, or by means of a multi-unit system such as a capsule containing sustained and immediate-release beads.

[0242] Sustained-release tablets containing hydrophilic polymers are prepared by techniques commonly known in the art, such as direct compression, wet granulation, or dry granulation processes. These formulations typically contain polymers, diluents, binders, lubricants, and active pharmaceutical ingredients. Common diluents include inert powdered substances such as starch, powdered cellulose, especially crystalline and microcrystalline cellulose, sugars such as fructose, mannitol, and sucrose, and cereal flour and similar edible powders. Typical diluents include, for example, various types of starch, inorganic salts such as lactose, mannitol, kaolin, calcium phosphate or calcium sulfate, and sodium chloride, and powdered sugars. Powdered cellulose derivatives are also useful. Typical tablet binders include substances such as starch, gelatin, and sugars such as lactose, fructose, and glucose. Natural and synthetic gums, including acacia, alginate, methylcellulose, and polyvinylpyrrolidone, can also be used. Polyethylene glycol, hydrophilic polymers, ethylcellulose, and waxes can also serve as binders. Lubricants are necessary in tablet formulations to prevent tablets and punches from sticking to the mold. Lubricants are selected from lubricating solids such as talc, magnesium and calcium stearate, stearic acid and hydrogenated vegetable oil.

[0243] Sustained-release tablets containing wax materials are generally prepared using methods known in the art, such as direct mixing, coagulation, and aqueous dispersion methods. In the coagulation method, the drug is mixed with the wax material, spray-coagulated or coagulated, sieved, and processed.

[0244] Delayed-release dosing regimen Delayed-release formulations can be produced by coating a solid dosage form with a polymer film that is insoluble in the acidic environment of the stomach and soluble in the neutral environment of the small intestine.

[0245] Delayed-release dose units can be prepared, for example, by coating a drug or drug-containing composition with a selected coating agent. The drug-containing composition may be, for example, a tablet for incorporation into a capsule, a tablet to be used as an inner core of a "coated core" dosage form, or a plurality of drug-containing beads, particles, or granules for incorporation into a tablet or capsule. Preferred coating agents may include biodegradable, gradually hydrolyzable, gradually water-soluble, and / or enzymatically soluble polymers, and may be conventional "enteric-coated" polymers. Enteric-coated polymers become soluble in the high-pH environment of the lower gastrointestinal tract or are slowly eroded as the dosage form passes through the gastrointestinal tract, as is recognized by those skilled in the art, while enzymatically soluble polymers are broken down by bacterial enzymes present in the lower gastrointestinal tract, particularly the colon. Suitable coating agents that provide delayed release include cellulose polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxymethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose acetate succinate, hydroxypropyl methylcellulose phthalate, methylcellulose, ethyl cellulose, cellulose acetate, cellulose acetate phthalate, cellulose acetate trimelliticate, and sodium carboxymethylcellulose; preferably acrylic acid polymers and copolymers and eudragit formed from acrylic acid, methacrylic acid, methyl acrylate, ethyl acrylate, methyl methacrylate, and / or ethyl methacrylate. (登録商標) L30D-55 and L100-55 (soluble at pH 5.5 or higher), Eudragit (登録商標) L-100 (soluble at pH 6.0 or higher), Eudragit (登録商標) S (soluble at pH 7.0 or higher as a result of advanced esterification) and Eudragit (登録商標) Trade name Eudragit, which includes NE, RL, and RS (water-insoluble polymers with varying degrees of permeability and expandability). (登録商標)Other methacrylic resins commercially available under Rohm Pharma (Westerstadt, Germany); vinyl polymers and copolymers such as polyvinylpyrrolidone, vinyl acetate, vinyl acetate phthalate, vinyl acetate crotonic acid copolymer and ethylene-vinyl acetate copolymer; enzymatically digestible polymers such as azopolymers, pectin, chitosan, amylose and guar gum; and zein and shellac, among others. Combinations of different coating agents may also be used. Multilayer coatings using various polymers may also be applied.

[0246] The preferred coating weight of a particular coating agent can be easily determined by those skilled in the art by evaluating the individual release profiles of tablets, beads, and granules prepared with different amounts of various coating agents. The combination of materials, methods, and application modes that produce the desired release characteristics can only be determined from clinical trials.

[0247] Coating compositions may contain conventional additives such as plasticizers, pigments, colorants, stabilizers, and flow enhancers. Plasticizers are usually present to reduce the brittleness of the coating and generally make up about 10% to 50% by weight of the dry weight of the polymer. Examples of typical plasticizers include polyethylene glycol, propylene glycol, triacetin, dimethyl phthalate, diethyl phthalate, dibutyl phthalate, dibutyl sebacate, triethyl citrate, tributyl citrate, triethyl acetyl citrate, castor oil, and acetylated monoglycerides. Stabilizers are preferably used to stabilize dispersed particles. Typical stabilizers are nonionic emulsifiers such as sorbitan esters, polysorbates, and polyvinylpyrrolidone. Flow enhancers are recommended to reduce tackiness during film formation and drying and generally make up about 25% to 100% by weight of the polymer in the coating solution. One effective flow enhancer is talc. Other flow enhancers, such as magnesium stearate and glycerol monostearate, may also be used. Dyes, such as titanium dioxide, may also be used. Small amounts of defoaming agents, such as silicone (e.g., simethicone), may also be added to the coating composition.

[0248] Preparations for lung and mucosal administration The activator and its composition may be formulated for pulmonary or mucosal administration. Administration includes delivery of the composition to the lungs, nasal, oral (sublingual, buccal), vaginal, or rectal mucosa.

[0249] In one embodiment, the compound is formulated for pulmonary delivery, such as intranasal administration or oral inhalation. The respiratory tract is a structure involved in gas exchange between the atmosphere and the bloodstream. The lungs are branched structures that ultimately end in the alveoli, where gas exchange takes place. The alveolar surface area is the largest in the respiratory system and is where drug absorption occurs. The alveoli are covered with thin epithelium that lacks a ciliary or mucous layer and secrete surfactant phospholipids. The respiratory tract includes the airways, which contain the oropharynx and larynx, followed by the airways, which contain the trachea, and then branch into the bronchi and bronchioles. The upper and lower airways are called the inductive airways. The terminal bronchioles then divide into respiratory bronchioles, which then lead to the respiratory region, alveoli, or deep lung. The deep lung or alveoli are the primary target of inhaled therapeutic aerosols for systemic drug delivery.

[0250] Lung administration of therapeutic compositions consisting of low molecular weight drugs has been observed, for example, beta-androgenic antagonists for treating asthma. Other therapeutic agents that are active in the lungs are administered systemically and targeted by pulmonary absorption. Nasal delivery is considered a promising technique for administering therapeutic agents for the following reasons: the nose has a large surface area available for drug absorption due to the epithelial surface coverage by numerous microvilli; the subepithelial layer is highly vascularized; venous blood from the nose passes directly into the systemic circulation, avoiding drug loss due to the first-pass effect in the liver; low doses are possible; therapeutic blood levels are achieved more rapidly; pharmacological activity develops quickly; and side effects are fewer. 3 The area has high total blood flow, a porous endothelial basement membrane, and is easily accessible.

[0251] The term "aerosol" as used herein refers to any formulation of fine mist of particles, whether produced using a propellant or otherwise, which may be a solution or suspension. Aerosols can be produced using standard techniques such as sonication or high-pressure processing.

[0252] Carriers for lung formulations can be divided into those for dry powder formulations and those for administration as solutions. Aerosols for the delivery of therapeutic agents into the respiratory tract are known in the art. For administration via the upper respiratory tract, formulations can be formulated into solutions, such as water or buffered or unbuffered isotonic saline or suspension, for intranasal administration as droplets or sprays. Preferably, such solutions or suspensions are isotonic with respect to nasal secretions and have approximately the same pH, for example, in the range of about pH 4.0 to about pH 7.4 or about pH 6.0 to about pH 7.0. The buffer must be physiologically compatible and includes, by example, phosphate buffer. For example, a typical nasal decongestant is described as being buffered to a pH of about 6.2. Those skilled in the art can easily determine the appropriate saline content and pH of a harmless aqueous solution for intranasal and / or upper respiratory administration.

[0253] Preferably, the aqueous solution is water, a physiologically acceptable aqueous solution containing salt and / or a buffer such as phosphate-buffered saline (PBS) or any other aqueous solution acceptable for administration to animals or humans. Such solutions are well known to those skilled in the art and include, but are not limited to, distilled water, deionized water, pure or ultrapure water, saline and PBS. Other suitable aqueous media include, but are not limited to, Ringer's solution and isotonic sodium chloride. The aqueous suspension may contain suspending agents such as cellulose derivatives, sodium alginate, polyvinylpyrrolidone and tragacanth gum and lecithin. Suitable preservatives for aqueous suspensions include ethyl p-hydroxybenzoate and n-propyl.

[0254] In other embodiments, solvents that are low-toxicity organic (i.e., non-aqueous) Class 3 residual solvents, such as ethanol, acetone, ethyl acetate, tetrahydrofuran, ethyl ether, and propanol, may be used in the formulation. The solvent is selected based on its ability to readily aerosolize the formulation. The solvent must not react toxicly with the compound. A suitable solvent that dissolves the compound or forms a suspension of the compound must be used. The solvent must be sufficiently volatile to allow the formation of an aerosol of the solution or suspension. Further solvents or aerosolizing agents, such as Freon, may be added if increased volatility of the solution or suspension is desired.

[0255] In one embodiment, the composition may contain trace amounts of polymers, surfactants, or other additives well known to those skilled in the art. In this context, “trace amounts” means that no additives are present that affect or mediate the uptake of the compound in the lungs, and that any additives present are not present in amounts that adversely affect the uptake of the compound in the lungs.

[0256] Due to their hydrophobic properties, dried lipid powders can be dispersed in ethanol. For lipids stored in an organic solvent such as chloroform, a desired amount of the solution is placed in a vial, and the chloroform is evaporated under a nitrogen stream to form a dry, thin film on the surface of the glass vial. When reconstituted with ethanol, the film expands readily. To completely distribute the lipid molecules in the organic solvent, the suspension is subjected to ultrasonic treatment. Non-aqueous suspensions of lipids can be prepared in anhydrous ethanol using a reusable PARI LC Jet+ nebulizer (PARI Respiratory Equipment, Monterey, CA).

[0257] Large particle size dry powder formulations ("DPFs") exhibit improved fluidity characteristics, including low aggregation, easy aerosolization, and possibly low phagocytosis. Dry powder aerosols for inhalation therapy generally produce an average diameter mainly in the range of less than 5 microns, although the preferred range is 1 to 10 microns in aerodynamic diameter. Large "carrier" particles (without drug) are co-delivered with the therapeutic aerosol to achieve sufficient aerosolization, although they offer several other benefits.

[0258] Polymer particles can be prepared using single and double emulsion solvent evaporation, spray drying, solvent extraction, solvent evaporation, phase separation, simple and complex coacervation, interfacial polymerization, and other methods well known to those skilled in the art. The particles can also be produced using methods for producing microspheres or microcapsules known in the art. Preferred production methods include spray drying and freeze-drying, which involve using a solution containing a surfactant, spraying it to form droplets of a desired size, and removing the solvent.

[0259] Particles can be manufactured with appropriate materials, surface roughness, diameter, and tap density for localized delivery to select areas of the respiratory tract, such as the deep lungs or upper airways. For example, high-density or large particles may be used for upper airway delivery. Similarly, mixtures of particles of various sizes with the same or different EGS may be administered in a single dose to target various areas of the lung.

[0260] Formulations for pulmonary delivery include monolayer phospholipid vesicles, liposomes, or lipoprotein particles. Such formulations containing nucleic acids and methods for producing them are well known to those skilled in the art. Liposomes contain phospholipids provided by various suppliers, including Avanti Polar Lipids, Inc. (Birmingham, Ala.). In one embodiment, the liposomes may contain ligand molecules specific to receptors on the surface of target cells in order to direct the liposomes toward the target cells.

[0261] 4. Transdermal Transdermal formulations can also be prepared. These are typically ointments, lotions, sprays, or patches, all of which can be prepared using standard technologies. Transdermal formulations may contain penetration enhancers.

[0262] IV. Combination Therapy The disclosed Akt3 modulator may be administered alone or in combination with one or more additional therapeutic agents to a subject requiring it. In one embodiment, the Akt3 modulator and the additional therapeutic agent are administered separately but simultaneously. The Akt3 modulator and the additional therapeutic agent may also be administered as part of the same composition. In another embodiment, the Akt3 modulator and the second therapeutic agent are administered separately at different time points but as part of the same treatment regimen.

[0263] The subject may be administered the first agent 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours or more, or 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days or more, before the administration of the second agent. In one embodiment, the subject may be administered the first agent once or more every 1, 2, 3, 4, 5, 6, 7, 14, 21, 28, 35, or 48 days before a single dose of the second agent. The Akt3 modulator may be either the first or second agent.

[0264] Akt3 modulators and additional therapeutic agents may be administered as part of a treatment regimen. For example, if the first therapeutic agent can be administered to the subject every four days, the second therapeutic agent may be administered on day 1, day 2, day 3, or day 4, or a combination thereof. The first or second therapeutic agent may be administered repeatedly throughout the entire treatment regimen.

[0265] Examples of molecules include cytokines, chemotherapeutic agents, radioisotopes, other immunotherapies, enzymes, antibiotics, antivirals (especially protease inhibitors alone or in combination with nucleosides for HIV or hepatitis B or C), antiparasitic agents (helminths, protozoa), growth factors, growth inhibitors, hormones, hormone antagonists, antibodies and their bioactive fragments (including humanized, single-chain, and chimeric antibodies), antigens and vaccine preparations (including adjuvants), peptide drugs, anti-inflammatory agents, ligands that bind to Toll-like receptors to activate the innate immune system (including, but not limited to, CpG oligonucleotides), molecules that mobilize and optimize the adaptive immune system, other molecules that activate or upregulate the action of cytotoxic T lymphocytes, NK cells, and helper T cells, and other molecules that inactivate or downregulate suppressor or regulatory T cells.

[0266] Further therapeutic agents are selected based on the condition, disorder, or disease being treated. For example, an Akt3 modulator may be co-administered with one or more additional agents that function to enhance or promote the immune response, or to reduce or inhibit the immune response.

[0267] A. Chemotherapy agents The disclosed Akt3 modulator may be combined with one or more chemotherapeutic agents or apoptosis promoters. Representative chemotherapeutic agents include amsacrin, bleomycin, busulfan, capecitabine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clofarabine, chrysanthaspase, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin, docetaxel, doxorubicin, epirubicin, etoposide, fludarabine, fluorouracil, gemcitabine, hydroxycarbamide, idarubicin, ifosfamide, irinotecan, leucovorin, liposomal doxorubicin, Liposomal daunorubicin, lomustine, melphalan, mercaptopurine, mesna, methotrexate, mitomycin, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, pentostatin, procarbazine, larcitrexed, satoraplatin, streptozocin, tegafur-uracil, temozolomide, teniposide, thiotepa, thioguanine, topotecan, treosulfan, vinblastine, vincristine, vindesine, vinorelbine, or combinations thereof. Representative apoptosis promoters include, but are not limited to, fludarabine, staurosporine, cycloheximide, actinomycin D, lactosylceramide, 15d-PGJ(2), and combinations thereof.

[0268] B. Anti-inflammatory drugs Other suitable therapeutic agents include, but are not limited to, anti-inflammatory agents. Anti-inflammatory agents may be non-steroidal, steroidal, or a combination thereof. One embodiment provides an oral composition containing about 1% (w / w) to about 5% (w / w), typically about 2.5% (w / w), of an anti-inflammatory agent. Representative examples of non-steroidal anti-inflammatory agents include oxicam, e.g., piroxicam, isoxicam, tenoxicam, sudoxicam; salicylic acids, e.g., aspirin, disalside, benolilate, trilysart, sappapyrine, sorprin, diflunisal, and fendosal; acetic acid derivatives, e.g., diclofenac, fenclofenac, indomethacin, sulindac, tolmetin, isoxepac, flofenac, thiopinac, didomethacin, acemetacin, fentiazac, zomepirac, clindanac, oxepinac, felbinac, and ketorolac; and fenametes, e.g., mefenam. , meclofenum, flufenum, niflum, and tolfenamic acid; propionic acid derivatives, such as ibuprofen, naproxen, benoxaprofen, flurbiprofen, ketoprofen, fenoprofen, fenbufen, indoprofen, pirprofen, carprofen, oxaprozin, pranoprofen, miroprofen, thioxaprofen, suprofen, aluminoprofen, and thiaprofen; pyrazoles, such as phenylbutazone, oxyfenbutazone, feprazone, azapropazone, and trimethazone, are included but not limited to these. Mixtures of these nonsteroidal anti-inflammatory drugs may also be used.

[0269] Representative examples of steroidal anti-inflammatory drugs include corticosteroids, such as hydrocortisone, hydroxyl-triamcinolone, alpha-methyldexamethasone, dexamethasone phosphate, beclomethasone dipropionate, clobetasol valerate, desonide, desoxymethasone, desoxycorticosterone acetate, dexamethasone, dichlorizone, diflorasone diacetate, diflucortolone valerate, fluadrenolone, fluchlorolone acetonide, fludrocortisone, flumetasone pivalate, fluocinolone acetonide, fluocinonide, flucortin butyl ester, fluocortolone, fluprednilidene acetate, fludrenolone, halcinonide, hydrocortisone acetate, hydrocortisone butyrate, methylprednisolone, and triamcinolone acetate. This includes, but is not limited to, nide, cortisone, cortodoxone, flucetonide, fludrocortisone, diflorazone diacetate, fluradrenolon, fludrocortisone, diflorazone diacetate, fluradrenolon acetonide, medrisone, amsinafel, amsinafid, betamethasone and its esters in balance, chloroprednisone, chloroprednisone acetate, crocoltrone, crescinolone, dichlorizone, difluprednate, fluchloronide, flunisolide, fluoromethalone, fluperolon, fluprednisolone, hydrocortisone valerate, hydrocortisone cyclopentylpropionate, hydrocortamate, meprednisone, paramethasone, prednisolone, prednisone, beclomethasone dipropionate, triamcinolone and mixtures thereof.

[0270] C. Immunosuppressants In some embodiments, the compounds disclosed herein reduce Treg activity or production. In some embodiments, the compounds disclosed herein are used in induction therapy for cancer. In some embodiments, the compounds disclosed herein are used in combination with other immunotherapies, immunomodulators, costimulatory activating agonists, other cytokines and chemokines and factors, vaccines, oncolytic viruses, cell therapies, small molecule and targeted therapies, chemotherapy and radiotherapy. In some embodiments, the immunomodulators include checkpoint inhibitors such as anti-PD1, anti-CTLA4, anti-TIM3, and anti-LAG3. In some embodiments, the costimulatory activating agonists include anti-OX40 and anti-GITR. In some embodiments, the cell therapies include engineered T cells, CAR-T cells, TCR-T cells, and others.

[0271] In one embodiment, the compounds disclosed herein are used in combination with other immunotherapies, immunomodulators, biological agents (e.g., antibodies), vaccines, small molecule and targeted therapies, anti-inflammatory agents, cell therapies (e.g., engineered Treg and other types of cells), chemotherapy and radiotherapy.

[0272] In one embodiment, the compounds disclosed herein are administered in vivo to a patient by intravenous, intramuscular, or other non-enteral means, either alone or in combination with other agents. They may also be administered intranasally, by inhalation, rectally, vaginally, topically, orally, or by implantation. In another embodiment, the compounds disclosed herein are applied ex vivo, either alone or in combination with other agents, to enhance the function of inhibitory Tregs, including native Tregs, induced Tregs, engineered Tregs, and other types of inhibitory T cells, which may then be used for treatment of the patient as desired.

[0273] In one embodiment, a further therapeutic agent is an immunosuppressant. The immunosuppressant is an antibody against other lymphocyte surface markers (e.g., CD40, alpha-4 integrin) or cytokines, or a fusion protein (e.g., CTLA-4-Ig(Orencia)). (登録商標)), TNFR-Ig(Enbrel (登録商標) )), TNF-α blockers, e.g., Enbrel, Remicade, Cimzia and Humira, cyclophosphamide (CTX) (i.e., Endoxan) (登録商標) , Citoxane (登録商標) Neosar (登録商標) Procytox (登録商標) Revimmune TM ), methotrexate (MTX) (i.e., rheumatoid arthritis) (登録商標) Trexall (登録商標) ), belimumab (e.g., Benlysta (登録商標) ) or other immunosuppressive drugs (e.g., cyclosporine A, FK506-like compounds, rapamycin compounds, or steroids), antiproliferative agents, cytotoxic agents, or other compounds that may aid in immunosuppression.

[0274] In one embodiment, a further therapeutic agent may be a checkpoint inhibitor. In one embodiment, the therapeutic agent may be a CTLA-4 fusion protein such as CTLA-4-Ig (abatacept). The CTLA-4-Ig fusion protein competes with the T cell co-stimulatory receptor, CD28, for binding to CD80 / CD86 (B7-1 / B7-2) in antigen-presenting cells, and therefore functions to inhibit T cell activation. In another embodiment, the therapeutic agent is a CTLA-4-Ig fusion protein known as beratacept. Beratacept contains two amino acid substitutions (L104E and A29Y) that can significantly increase avidity to CD86 in vivo. In yet another embodiment, the therapeutic agent is Maxy-4.

[0275] In another embodiment, the therapeutic agent is cyclophosphamide (CTX). (登録商標) , Citoxane (登録商標) Neosar (登録商標) Procytox (登録商標) and Revimmune TM Its generic name is also known as cytophosphan, and it is a nitrogen mustard alkylating agent of the oxazofolin group.

[0276] Therapeutic agents may be administered to patients in need in an effective dose that reduces blood or serum levels of anti-double-stranded DNA (anti-ds DNA) autoantibodies and / or reduces proteinuria.

[0277] In other embodiments, the therapeutic agent increases the amount of adenosine in the serum (see, for example, WO08 / 147482, which is incorporated herein by reference in whole). For example, the second therapeutic agent may be CD73-Ig, recombinant CD73, or other agents that increase the expression of CD73 (e.g., cytokines, monoclonal antibodies, or small molecules) (see, for example, WO04 / 084933, which is incorporated herein by reference in whole). In other embodiments, the therapeutic agent is interferon-beta.

[0278] The therapeutic agent may be a small molecule that inhibits or reduces the differentiation, proliferation, activity and / or cytokine production and / or secretion by other cells, including but not limited to Th1, Th17, Th22 and / or IL-1β, TNF-α, TGF-beta, IFN-γ, IL-18, IL-17, IL-6, IL-23, IL-22, IL-21 and MMPs, or causes other cells to secrete inflammatory molecules. In other embodiments, the therapeutic agent is a small molecule that interacts with Tregs, enhances Treg activity, promotes or enhances IL-10 secretion by Tregs, increases the number of Tregs, increases the inhibitory performance of Tregs, or a combination thereof.

[0279] In one embodiment, the composition increases Treg activity or production. Examples of Treg enhancers include, but are not limited to, the glucocorticoid fluticasone, salmeterol, antibodies against IL-12, IFN-γ, and IL-4; vitamin D3 and dexamethasone and combinations thereof.

[0280] In one embodiment, the therapeutic agent is an antibody against pro-inflammatory molecules such as IL-6, IL-23, IL-22, or IL-21, for example, a functional blockade antibody.

[0281] The term "rapamycin compound" as used herein includes the neutral tricyclic compound rapamycin, rapamycin derivatives, rapamycin analogs, and other macrolide compounds thought to have the same mechanism of action as rapamycin (e.g., cytokine function inhibition). The term "rapamycin compound" also includes compounds structurally similar to rapamycin, such as compounds with similar macrocyclic structures that have been modified to enhance therapeutic efficacy. Examples of rapamycin compounds are known in the art (see, for example, WO95122972; WO95116691; WO95104738; U.S. Patents 6,015,809; 5,989,591; 5,567,709; 5,559,112; 5,530,006; 5,484,790; 5,385,908; 5,202,332; 5,162,333; 5,780,462; 5,120,727; each is incorporated herein by reference in whole).

[0282] The term "FK506-like compound" includes FK506 and FK506 derivatives and analogs, such as compounds structurally similar to FK506, for example, compounds having a similar macrocyclic structure that has been modified to enhance therapeutic efficacy. Examples of FK506-like compounds include those described in WO00101385 (which is incorporated herein by reference in its entirety). In some embodiments, the term "rapamycin compound" as used herein does not include FK506-like compounds.

[0283] D. Treatment of neurodegenerative diseases The disclosed Akt3 modulator may be administered in conjunction with a second treatment selected based on the disease state of the subject. The second treatment may be a treatment for Alzheimer's disease. Current treatments for Alzheimer's disease include, but are not limited to, cholinesterase inhibitors, e.g., donepezil, rivastigmine, and galantamine; memantine; antidepressants, e.g., citalopram, fluoxetine, paroxetine, sertraline, and trazodone; anxiolytics, e.g., lorazepam and oxazepam; and antipsychotics, e.g., aripiprazole, clozapine, haloperidol, olanzapine, quetiapine, risperidone, and ziprasidone.

[0284] In other embodiments, further therapeutic agents may be used to treat ALS. There are currently two US FDA-approved treatments for ALS: riluzole and edaravone. Both drugs have been shown to slow the progression of ALS. In addition to riluzole and edaravone, subjects with ALS may also be treated with drugs that target specific symptoms of the disease. Examples of drugs include, but are not limited to, drugs that reduce spasticity, such as antispasmodics (e.g., baclofen, dantrolene, and diazepam); drugs that help manage neuropathic pain, such as amitriptyline, carbamazepine, duloxetine, gabapentin, lamotrigine, milnacipran, nortriptyline, pregabalin, and venlafexine; and drugs that help patients swallow, such as trihexyphenidyl or amitriptyline.

[0285] In one embodiment, further therapeutic agents may be used for the treatment of Parkinson's disease. Current treatments for Parkinson's disease include, but are not limited to, carbidopa-levodopa; dopamine agonists such as pramipexole, ropinirole, and rotigotine; MAO B inhibitors such as selegiline, rasagiline, and safinamide; catechol O-methyltransferase inhibitors such as entacapone and tolcapone; anticholinergics such as benztropine and trihexyphenidyl; and amantadine.

[0286] A second treatment may be used for the treatment of Huntington's disease. Current treatments for Huntington's disease include, but are not limited to, tetrabenazine; antipsychotics such as haloperidol, chlorpromazine, risperidone, and quetiapine; antidepressants such as amantadine; levetiracetam; clonazepam; citalopram, escitalopram, fluoxetine, and sertraline; and anticonvulsants such as valoproate, carbamazepine, and lamotrigine.

[0287] E. Treatment for weight loss In one embodiment, the disclosed Akt3 modulator may be administered to the subject together with further therapeutic agents used to treat cachexia or extreme weight loss. The current strategy for treating cachexia and extreme weight loss is appetite enhancement using appetite stimulants to ensure adequate nutritional intake. Pharmacological interventions with appetite stimulants, nutritional supplements, 5-HT3 antagonists, and Cox-2 inhibitors have been used to treat cancer cachexia.

[0288] In one embodiment, the appetite stimulant is a vitamin, mineral, or herb, including but not limited to zinc, thiamine, or fish oil. In another embodiment, the appetite enhancer is a pharmaceutical, including but not limited to dronabinol, megestrol, and oxandrolone.

[0289] In the foregoing specification, the present invention is described in relation to one embodiment thereof, and many details are described for illustrative purposes. However, it will be recognized by those skilled in the art that the present invention is open to further embodiments, and some of the details described herein can be considerably modified without departing from the fundamental principles of the present invention.

[0290] All references cited herein are incorporated herein by whole-word by reference. The present invention can be embodied in other concrete forms without departing from its spirit or essential characteristics, and therefore, the scope of the invention should be described by the appended claims rather than the foregoing specification.

Claims

1. A method for treating a disease in a subject requiring treatment, comprising administering to the subject a composition comprising an Akt3 modulator in an amount effective to modulate Akt3 signaling and to treat or delay the progression of the disease.

2. The method of claim 1, wherein the disease is selected from the group consisting of neurodegenerative diseases, cachexia, eating disorders, obesity complications, inflammatory diseases, virus-induced inflammatory responses, Gulf War syndrome, tuberous sclerosis, retinitis pigmentosa, graft rejection, cancer, ischemic tissue injury, traumatic tissue injury, and combinations thereof.

3. The method according to claim 2, wherein the disease is a neurodegenerative disease.

4. The method of claim 3, wherein the neurodegenerative disease is selected from the group consisting of Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, motor neuron disorders, Huntington's disease, HIV-induced neurodegeneration, Lewy body disease, spinal muscular atrophy, prion disease, spinocerebellar ataxia, familial amyloid polyneuropathy, and combinations thereof.

5. The method of claim 2, wherein the disorder is cachexia or an eating disorder.

6. The method of claim 2, wherein the disease is a complication of obesity.

7. The method of claim 6, wherein the obesity complication is selected from the group consisting of glucose intolerance, fatty liver, dyslipidemia, and combinations thereof.

8. The method of claim 2, wherein the disease is an inflammatory disease.

9. The method of claim 8, wherein the inflammatory disease is selected from the group consisting of atopic dermatitis, allergies, asthma, and combinations thereof.

10. The method of claim 2, wherein the disease is a virus-induced inflammatory response.

11. The method of claim 10, wherein the virus-induced inflammatory response is SARS-induced inflammatory pneumonia, coronavirus disease 2019, or a combination thereof.

12. The method of claim 2, wherein the disease is Gulf War Syndrome or tuberous sclerosis.

13. The method of claim 2, wherein the disease is retinitis pigmentosa or graft rejection.

14. The method according to claim 2, wherein the disease is ischemic tissue injury or traumatic tissue injury.

15. The method of claim 2, wherein the disease is cancer.

16. The method of claim 15, wherein the cancer is selected from the group consisting of adult T-cell leukemia / lymphoma, bladder, brain, breast, cervix, colorectal, esophagus, kidney, liver, lung, nasopharyngeal, pancreatic, prostate, skin, stomach, uterus, ovaries, and testes.

17. The method of claim 15, wherein the cancer is leukemia.

18. The method of claim 17, wherein the leukemia is adult T-cell leukemia / lymphoma.

19. The method according to claim 18, wherein adult T-cell leukemia / lymphoma is caused by a human T-cell lymphotropic virus.

20. A method according to any one of claims 1 to 19, wherein Akt3 is regulated by immune cells.

21. The method of claim 20, wherein the immune cells are selected from the group consisting of T cells, B cells, macrophages, and glial cells.

22. The method of claim 21, wherein the glial cells are astrocytes, microglia, or oligodendrocytes.

23. The method according to claim 21, wherein the T cells are T regulatory cells.

24. The method according to claim 1 or 2, wherein an Akt3 modulator activates Akt3 signaling.

25. The method according to claim 1 or 2, wherein an Akt3 modulator inhibits Akt3 signaling.

26. The method according to claim 1 or 2, wherein the Akt3 modulator increases T-regulatory cell activity or production.

27. The method according to claim 1 or 2, wherein the Akt3 modulator reduces T regulatory cell activity or production.

28. The modulator of Akt3 is given by equation I: 【Chemistry 1】 A compound of or a pharmaceutically acceptable enantiomer, salt, or solvate thereof, where: Rings A, B, and C are independently six-membered aryl or N-containing heteroaryl monocyclic or bicyclic ring systems containing zero or more N atoms, selected from the group consisting of phenyl, pyridine, pyrimidine, pyridazine, pyrazine, triazine, quinoline, quinazoline, isoquinoline, naphthalene, naphthyridine, indole, isoindole, cinnoline, phthalazine, quinoxaline, pteridine, purine, and benzimidazole; R １ is optionally -(C １ -C １２ )alkyl, -(C ３ -C １２ )cycloalkyl, -(C ３ -C １２ )heterocycloalkyl, -O-(C １ -C １２ )alkyl, -O-(C １ -C １２ )alkyl-(C ６ -C ２０ )aryl, -O-(C ３ -C １２ )cycloalkyl, -S-(C １ -C １２ )alkyl, -S-(C ３ -C １２ )cycloalkyl, -COO-(C １ -C １２ )alkyl, -COO-(C ３ -C １２ )cycloalkyl, -CONH-(C １ -C １２ )alkyl, -CONH-(C ３ -C １２ )cycloalkyl, -CO-(C １ -C １２ )alkyl, -CO-(C ３ -C １２ )cycloalkyl, -N-[(C １ -C １２ )alkyl] ２ , -(C ６ -C ２０ )aryl, -(C ６ -C ２０ )aryl-(C １ -C １２ )alkyl, -(C ６ -C ２０ )aryl-O-(C １ -C １２ )alkyl, -(C ３ -C ２０ )heteroaryl, -(C ３ -C ２０ )heteroaryl-(C １ -C １２ )-alkyl,-(C ３ -C ２０ )-heteroaryl-O-(C １ -C １２ )-alkyl,-COOH,-OH,-SH,-SO ３ H, -CN, -NH ２ or substituted with one or more substituents selected from halogens - (C １ -C ３０ )-alkyl,-(C ３ -C １２ )-cycloalkyl,-(C ３ -C １２ )-heterocycloalkyl,-(C ６ -C ２０ )-aryl or-(C ３ -C ２０ ) - It is a heteroaryl group; X, Y, and Z are independent of each other = O, -NH, -S, -N-(C １ -C ３０ )-alkyl or-(C １ -C ３０ ) - is aryl; 【Chemistry 2】 ga-CH((C １ -C ３０ )-alkyl))-, -(C=O)-, -CH(OH), -SO ２ -, -SO- or -CH(SOCH ３ ) - and; and R ３ optionally -(C １ -C １２ )-alkyl, -(C ３ -C １２ )-cycloalkyl, -(C ３ -C １２ )-heterocycloalkyl, -O-(C １ -C １２ )-alkyl, -O-(C １ -C １２ )-alkyl-(C ６ -C ２０ )-aryl, -O-(C ３ -C １２ )-cycloalkyl, -S-(C １ -C １２ )-alkyl, -S-(C ３ -C １２ )-cycloalkyl, -COO-(C １ -C １２ )-alkyl, -COO-(C ３ -C １２ )-cycloalkyl, -CONH-(C １ -C １２ )-alkyl, -CONH-(C ３ -C １２ )-cycloalkyl, -CO-(C １ -C １２ )-alkyl, -CO-(C ３ -C １２ )-cycloalkyl, -N-[(C １ -C １２ )-alkyl] ２ , -(C ６ -C ２０ )-aryl, -(C ６ -C ２０ )-aryl-(C １ -C １２ )-alkyl, -(C ６ -C ２０ )-aryl-O-(C １ -C １２ )-alkyl, -(C ３ -C ２０ )-heteroaryl, -(C ３ -C ２０ )-heteroaryl-(C １ -C １２ )-alkyl,-(C ３ -C ２０ )-heteroaryl-O-(C １ -C １２ )-alkyl,-COOH,-OH,-SH,-SO ３ H, -CN, -NH ２ or substituted with one or more substituents selected from halogens - (C １ -C ３０ )-alkyl,-(C ３ -C １２ )-cycloalkyl,-(C ３ -C １２ )-heterocycloalkyl,-(C ６ -C ２０ )-aryl or-(C ３ -C ２０ ) - Heteroaryl group, The method according to any one of claims 1 to 27.

29. The Akt3 modulator is given by equation II: 【Transformation 3】 A compound of or a pharmaceutically acceptable enantiomer, salt, or solvate thereof, where: R １ Depending on the case - (C １ -C １２ )-alkyl,-(C ３ -C １２ )-cycloalkyl,-(C ３ -C １２ )-heterocycloalkyl, -O-(C １ -C １２ )-alkyl,-O-(C １ -C １２ )-alkyl-(C ６ -C ２０ )-aryl,-O-(C ３ -C １２ )-cycloalkyl,-S-(C １ -C １２ )-alkyl,-S-(C ３ -C １２ )-cycloalkyl,-COO-(C １ -C １２ )-alkyl,-COO-(C ３ -C １２ )-cycloalkyl,-CONH-(C １ -C １２ )-alkyl,-CONH-(C ３ -C １２ )-cycloalkyl,-CO-(C １ -C １２ )-alkyl,-CO-(C ３ -C １２ )-cycloalkyl,-N-[(C １ -C １２ )-alkyl] ２ , -(C ６ -C ２０ )-Aryl,-(C ６ -C ２０ )-Aryl-(C １ -C １２ )-alkyl,-(C ６ -C ２０ )-aryl-O-(C １ -C １２ )-alkyl,-(C ３ -C ２０ )-heteroaryl,-(C ３ -C ２０ )-heteroaryl-(C １ -C １２ )-alkyl,-(C ３ -C ２０ )-heteroaryl-O-(C １ -C １２ )-alkyl,-COOH,-OH,-SH,-SO ３ H, -CN, -NH ２ or substituted with one or more substituents selected from halogens - (C １ -C ３０ )-alkyl,-(C ３ -C １２ )-cycloalkyl,-(C ３ -C １２ )-heterocycloalkyl,-(C ６ -C ２０ )-aryl or-(C ３ -C ２０ ) - It is a heteroaryl group; X, Y, and Z are independently -O, -NH, -S, -N-(C １ -C ３０ )-alkyl or-(C １ -C ３０ ) - is aryl; 【Chemistry 4】 ga-CH((C １ -C ３０ )-alkyl))-, -(C=O)-, -CH(OH), -SO ２ -, -SO- or -CH(SOCH ３ ) - and; and R ３ Depending on the case - (C １ -C １２ )-alkyl,-(C ３ -C １２ )-cycloalkyl,-(C ３ -C １２ )-heterocycloalkyl, -O-(C １ -C １２ )-alkyl,-O-(C １ -C １２ )-alkyl-(C ６ -C ２０ )-aryl,-O-(C ３ -C １２ )-cycloalkyl,-S-(C １ -C １２ )-alkyl,-S-(C ３ -C １２ )-cycloalkyl,-COO-(C １ -C １２ )-alkyl,-COO-(C ３ -C １２ )-cycloalkyl,-CONH-(C １ -C １２ )-alkyl,-CONH-(C ３ -C １２ )-cycloalkyl,-CO-(C １ -C １２ )-alkyl,-CO-(C ３ -C １２ )-cycloalkyl,-N-[(C １ -C １２ )-alkyl] ２ , -(C ６ -C ２０ )-Aryl,-(C ６ -C ２０ )-Aryl-(C １ -C １２ )-alkyl,-(C ６ -C ２０ )-aryl-O-(C １ -C １２ )-alkyl,-(C ３ -C ２０ )-heteroaryl,-(C ３ -C ２０ )-heteroaryl-(C １ -C １２ )-alkyl,-(C ３ -C ２０ )-heteroaryl-O-(C １ -C １２ )-alkyl,-COOH,-OH,-SH,-SO ３ H, -CN, -NH ２ or substituted with one or more substituents selected from halogens - (C １ -C ３０ )-alkyl,-(C ３ -C １２ )-cycloalkyl,-(C ３ -C １２ )-heterocycloalkyl,-(C ６ -C ２０ )-aryl or-(C ３ -C ２０ ) - Heteroaryl group, The method according to any one of claims 1 to 28.

30. The Akt3 modulator is given by equation III: 【Transformation 5】 A compound of or a pharmaceutically acceptable enantiomer, salt, or solvate thereof, where: R １ Depending on the case - (C １ -C １２ )-alkyl,-(C ３ -C １２ )-cycloalkyl,-(C ３ -C １２ )-heterocycloalkyl, -O-(C １ -C １２ )-alkyl,-O-(C １ -C １２ )-alkyl-(C ６ -C ２０ )-aryl,-O-(C ３ -C １２ )-cycloalkyl,-S-(C １ -C １２ )-alkyl,-S-(C ３ -C １２ )-cycloalkyl,-COO-(C １ -C １２ )-alkyl,-COO-(C ３ -C １２ )-cycloalkyl,-CONH-(C １ -C １２ )-alkyl,-CONH-(C ３ -C １２ )-cycloalkyl,-CO-(C １ -C １２ )-alkyl,-CO-(C ３ -C １２ )-cycloalkyl,-N-[(C １ -C １２ )-alkyl] ２ , -(C ６ -C ２０ )-Aryl,-(C ６ -C ２０ )-Aryl-(C １ -C １２ )-alkyl,-(C ６ -C ２０ )-aryl-O-(C １ -C １２ )-alkyl,-(C ３ -C ２０ )-heteroaryl,-(C ３ -C ２０ )-heteroaryl-(C １ -C １２ )-alkyl,-(C ３ -C ２０ )-heteroaryl-O-(C １ -C １２ )-alkyl,-COOH,-OH,-SH,-SO ３ H, -CN, -NH ２ or substituted with one or more substituents selected from halogens - (C １ -C ３０ )-alkyl,-(C ３ -C １２ )-cycloalkyl,-(C ３ -C １２ )-heterocycloalkyl,-(C ６ -C ２０ )-aryl or-(C ３ -C ２０ ) - It is a heteroaryl group; X, Y, and Z are independently -O, -NH, -S, -N-(C １ -C ３０ )-alkyl or-(C １ -C ３０ ) - is aryl; 【Transformation 6】 ga-CH((C １ -C ３０ )-alkyl))-, -(C=O)-, -CH(OH), -SO ２ -, -SO- or -CH(SOCH ３ ) - and; and R ４ ga-(C １ -C １２ )-alkyl,-(C ３ -C １２ )-cycloalkyl,-(C ３ -C １２ )-heterocycloalkyl, -O-(C １ -C １２ )-alkyl,-O-(C １ -C １２ )-alkyl-(C ６ -C ２０ )-aryl,-O-(C ３ -C １２ )-cycloalkyl,-S-(C １ -C １２ )-alkyl,-S-(C ３ -C １２ )-cycloalkyl,-COO-(C １ -C １２ )-alkyl,-COO-(C ３ -C １２ )-cycloalkyl,-CONH-(C １ -C １２ )-alkyl,-CONH-(C ３ -C １２ )-cycloalkyl,-CO-(C １ -C １２ )-alkyl,-CO-(C ３ -C １２ )-cycloalkyl,-N-[(C １ -C １２ )-alkyl] ２ , -(C ６ -C ２０ )-Aryl,-(C ６ -C ２０ )-Aryl-(C １ -C １２ )-alkyl,-(C ６ -C ２０ )-aryl-O-(C １ -C １２ )-alkyl,-(C ３ -C ２０ )-heteroaryl,-(C ３ -C ２０ )-heteroaryl-(C １ -C １２ )-alkyl,-(C ３ -C ２０ )-heteroaryl-O-(C １ -C １２ )-alkyl,-COOH,-OH,-SH,-SO ３ H, -CN, -NH ２ or halogen, The method according to any one of claims 1 to 28.

31. The Akt3 modulator is given by equation IV: 【Transformation 7】 The method according to any one of claims 1 to 28, wherein the compound or a pharmaceutically acceptable enantiomer, salt, or solvate thereof.

32. The method according to any one of claims 1 to 31, further comprising administering a second therapeutic agent to the subject.

33. The method of claim 32, wherein the second therapeutic agent is selected from the group consisting of nutritional supplements, chemotherapeutic agents, anti-inflammatory agents, immunosuppressants, cholinesterase inhibitors, antidepressants, anxiolytics, antipsychotics, riluzole, edaravone, dopamine agonists, MAO B inhibitors, catechol O-methyltransferase inhibitors, anticholinergics, anticonvulsants, tetrabenazine, carbidopa-levodopa, antispasmodics, antibodies, fusion proteins, enzymes, nucleic acids, ribonucleic acids, antiproliferative agents, cytotoxic agents, appetite stimulants, 5-HT3 antagonists, Cox-2 inhibitors, and combinations thereof.

34. A method for treating cachexy in a subject requiring treatment, comprising administering to the subject a composition containing a selective Akt3 inhibitor in an effective amount that inhibits Akt3 signaling in adipocytes and activates adipogenesis.

35. The method of claim 34, further comprising administering a second therapeutic agent to the subject.

36. The method of claim 35, wherein the second therapeutic agent is selected from the group consisting of appetite stimulants, nutritional supplements, 5-HT3 antagonists, Cox-2 inhibitors, chemotherapeutic agents, anti-inflammatory agents, immunosuppressants, cholinesterase inhibitors, antidepressants, anxiolytics, antipsychotics, riluzole, edaravone, dopamine agonists, MAO B inhibitors, catechol O-methyltransferase inhibitors, anticholinergics, anticonvulsants, tetrabenazine, carbidopa-levodopa, antispasmodics, antibodies, fusion proteins, enzymes, nucleic acids, ribonucleic acids, antiproliferative agents, cytotoxic agents, and combinations thereof.

37. The method of claim 36, wherein the second therapeutic agent is an appetite stimulant, a nutritional supplement, a 5-HT3 antagonist, or a Cox-2 inhibitor.

38. The method according to any one of claims 34 to 37, wherein the target is neurodegenerative disease, cachexia, eating disorders, obesity complications, inflammatory diseases, virus-induced inflammatory responses, Gulf War syndrome, tuberous sclerosis, retinitis pigmentosa, graft rejection, cancer, and combinations thereof.

39. The method according to any one of claims 1 to 38, wherein the Akt3 inhibitor is a compound selected from the group consisting of the following: 【Transformation 8】 【Chemistry 9】

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