RU2815372C2 - Specific activator akt3 and its use - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to a compound of formula I or a pharmaceutically acceptable salt thereof, wherein rings A, B and C are independently phenyl, pyridine and quinoline; R₁ represents -(C₃-C₁₂)-heterocycloalkyl, -(C₆-C₂₀)-aryl, or -(C₃-C₂₀)-heteroaryl groups, optionally substituted by one or more substituents selected from - (C₁-C₁₂)-alkyl or -NH₂; X, Y and Z are independently -O, -NH, or -N-(C₁-C₃₀)-alkyl; R₂ represents =O; and R₃ on ring A is -(C₁-C₃₀)-alkyl, -(C₃-C₁₂)-heterocycloalkyl, -(C₆-C₂₀)-aryl, -O-(C₁-C₁₂)-alkyl, -N-[(C₁-C₁₂)-alkyl]₂, -COOH, -CN, -NH₂ or halogen; R₃ on ring C is hydrogen, -(C₁-C₃₀)-alkyl or halogen; provided that when R₁ is unsubstituted pyridine or N-substituted –(C1)-alkyl pyridine, X, Y and Z are NH, R₂ is =O, A is quinoline or pyridine, B is phenyl or pyridine, C is phenyl, R₃ on ring C is hydrogen, then R₃ on ring A is not -N-[(C₁)-alkyl]₂, -COOH, -CN and –F; when the compound has the structure of formula III

Formula III

R₁ is unsubstituted pyridine, N-substituted -(C₁-C₃)-alkyl pyridine or pyrimidine substituted –(C₁)-alkyl and NH₂, X, Y, Z are -NH, R₂ is =O, then R₄ is not a halogen, -(C₁)-alkyl, -O-(C₁)-alkyl, -NH₂ and -N-[(C₁)-alkyl]₂; when R₁ is unsubstituted pyridine, unsubstituted quinoline or quinoline substituted with -NH₂, -NO₂, or -N-[(C₁)-alkyl]₂, X, Y, Z are -NH, R₂ is =O, A is pyridine, B is phenyl, C is phenyl, R₃ on ring C is hydrogen, then R₃ on ring A is not –(C₁-C₂)-alkyl and -NH₂; and when R₁ is acridine, X, Y, Z are -NH, R₂ is =O, A is pyridine, B is phenyl, C is phenyl, R₃ on ring C is hydrogen, then R₃ on ring A is not –(C₁)-alkyl. The invention also relates to pharmaceutical compositions and methods for selective activation of Akt3.

Formula I

EFFECT: new compounds that can be used in medicine for the effective treatment of inflammatory, autoimmune diseases, graft-versus-host disease, chronic infection and obesity.

30 cl, 112 dwg, 13 ex

Description

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims benefit and priority to US Provisional Application No. 62/555141, filed September 7, 2017, 62/657345, filed April 13, 2018, and 62/659870, filed April 19, 2018, the contents of which are incorporated herein by reference. in full.

LINK TO LIST OF SEQUENCES

The sequence listing, filed on September 6, 2018, in the form of a text file named "064466.070 sequence listing_ST25.txt", which was created on August 21, 2018 and is 10.7 kilobytes in size, is incorporated herein by reference in accordance with 37 C.F.R. § 1.52(e)(5).

TECHNICAL FIELD

This invention is generally directed to compositions and methods for selectively activating Akt3 activity and methods of using them to modulate regulatory T cells.

BACKGROUND OF THE ART

Regulatory T cells (Treg) are a subset of CD4+ T cells that suppress immune responses and are important mediators of autotolerance and immune homeostasis (Sakaguchi, et al., Cell , 133, 775-787 (2008)). Depletion or inactivation of Tregs leads to severe autoimmune reactions (Sakaguchi, et al., J. Immunol. , 155, 1151-1164 (1995)), and their accumulation inhibits antitumor immunity (Dannull, et al., The Journal of clinical investigation , 115, 3623-3633 (2005)). Tregs are characterized by the expression of Foxp3, which is a transcription factor belonging to the Forkhead Box family of transcription factors. Foxp3 is a master regulator of Treg cells as it is essential for their development and function (Hori, Science , 299, 1057-1061 (2003); Fontenot, et al., Nat Immunol. , 4(4):330-6 (2003) Epub 2003 Mar 3; Khattri, et al., Nat Immunol. , 4(4):337-42 (2003). Epub 2003 Mar 3).

There are two main types of Treg cells: thymus-derived Tregs (or natural Tregs (nTregs)), which constitute 5-10% of total peripheral CD4+ T cells, and peripheral TGFβ-induced Tregs (iTregs). Both types have been demonstrated to have immunosuppressive properties mediated by several processes involving immunosuppressive soluble factors or cell contact (Bluestone, et al., Nat Rev Immunol , 3, 253-257 (2003); Glisic, et al., Cell and Tissue Research , 339, 585-595 (2010); Hori, Science , 299, 1057-1061 (2003); Sakaguchi, Cell , 101, 455-458 (2000); Sakagushi, et al., Curr. Top Microbiol. Immunol . , 305, 51-66 (2006); Sakagushi, et al., Immunol. , Rev. , 212, 8-27 (2006); (Schmidt, et al., Front Immunol. , 3:51 (2012)) However, the molecular mechanisms by which nTregs and iTregs develop and then exhibit nonredundant immune suppressive properties are not fully understood (Dipica, et al., Immunity , 35 (1): 109-122 (2011)).

PI3K-Akt signaling influences many processes and is central to many signaling pathways. Akt phosphorylation and kinase activity are induced by PI3K activation, which in turn is induced by several growth factor receptors, TCR, CD28 and IL-2R, among many others (Parry, et al., Trends in Immunology , 28, 161-168 (2007 )). Mammals have three isoforms of Akt, namely Akt1, Akt2 and Akt3, encoded by three independent genes. In vitro , these isoforms appear to have redundant functions, since different extracellular inputs can induce similar Akt signaling profiles (Franke, Science 1, pe29- (2008)). However, isoform-specific knockouts exhibit unique features and their involvement in disease and physiological conditions differs (Boland, et al., American Journal of Human Genetics , 81, 292-303 (2007); DeBosch, et al., J. Biol. Chem , 281, 32841-32851 (2006); Emamian, et al., Nat Genet , 36, 131-137 (2004); Garofalo, et al., The Journal of clinical investigation , 112, 197-208 (2003); George, et al., Science , 304, 1325-1328 (2004); Nakatani, et al., The Journal of Biological Chemistry , 274, 21528-21532 (1999); Tschopp, et al., Development ( Cambridge, England , 132, 2943-2954 (2005); Yang, et al., J. Biol. Chem. , 278, 32124-32131 (2003)).

Studies have demonstrated that Akt1 and Akt2 can negatively regulate characteristic transcriptional signatures of Treg, thereby selectively influencing Treg lineage differentiation (Sauer, et al., Proceedings of the National Academy of Sciences , 105, 7797-7802 (2008a)). Additionally, although inhibition of Akt1 and Akt2 isoforms has been shown to increase Foxp3 expression in TGFβ-induced iTregs (Sauer, et al., Proc. Natl. Acad. Sci. USA , 105, 7797-7802 (2008b)) , this mechanism remains unclear. Another finding demonstrates that Akt2 deletion resulted in defective differentiation of iTh17 cells but spared nTh17 cell development (Kim, et al., Nat Immunol ., 14 (6): 611-8 (2013) Epub 2013 May 5). In addition, Akt3 is also expressed in immune cells and in the spinal cord of Akt3 knockout mice, which have reduced numbers of Foxp3+ regulatory T cells compared to wild-type mice (Tsiperson, et al., J Immunol. , 190(4) :1528-39 (2013) Epub 2013 Jan 18)). Thus, although several studies have examined the significance of Akt isoform expression in T cell biology (Carson, et al., Annals of the New York Academy of Sciences , 1103, 167-178 (2007), Crellin, et al. ., Blood , 109, 2014-2022 (2007a); Crellin, et al., Journal of Immunological Methods , 324, 92-104 (2007b); Haxhinasto, J. Exp. Med. , 205, 565-574 (2008) Li, et al., Blood , 106, 3068-3073 (2005); Patton, et al., Biochem. Soc. Trans. , 35, 167-171 (2007); Patton, et al., J. Immunology 177 , 6598-6602 (2006); Sauer, et al., Proc. Natl. Acad. Sci. USA , 105, 7797-7802 (2008b); Walsh, et al., J. Clin. Invest. , 116, 2521- 2531. (2006)), the roles that Akt isoforms play in Treg function and induction still remain unclear.

Therefore, the purpose of the invention is to provide compounds and compositions for selective activation of Akt3 in immune cells.

Another object of the invention is to provide methods for reducing the immune response in a subject.

It is yet another object of the invention to provide methods for enhancing a suppressive immune response in a subject.

SUMMARY OF THE INVENTION

The present invention provides compositions and methods for selectively activating Akt3. In one embodiment of the present invention there is provided a compound according to Formula I:

or a pharmaceutically acceptable enantiomer, salt or solvate thereof, wherein:

rings A, B and C are independently six-membered aryl or N-containing heteroaryl mono- 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.

R ₁ is selected from -(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 groups optionally substituted with one or more substituents selected from -(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;

X, Y and Z are independently selected from =O, -NH, -S, -N-(C ₁ -C ₃₀ )-alkyl or -(C ₁ -C ₃₀ )-aryl;

R ₂ is selected from -(C ₁ -C ₃₀ )-alkyl, =O, -OH, -SO ₂ , -SO, or -SOCH ₃ ; And

R ₃ is selected from -(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 groups, optionally substituted by one or more substituents selected from -(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.

In another embodiment of the present invention there is provided a compound according to formula II:

or a pharmaceutically acceptable enantiomer, salt or solvate thereof, wherein:

R ₁ is selected from -(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 groups optionally substituted with one or more substituents selected from -(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;

X, Y and Z are independently selected from -O, -NH, -S, -N-(C ₁ -C ₃₀ )-alkyl or -(C ₁ -C ₃₀ )-aryl;

R ₂ is selected from -(C ₁ -C ₃₀ )-alkyl, -O, -OH, -SO ₂ , -SO, or -SOCH ₃ ; And

R ₃ is selected from -(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 groups, optionally substituted by one or more substituents selected from -(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.

In another embodiment of the present invention there is provided a compound according to formula III:

or a pharmaceutically acceptable enantiomer, salt or solvate thereof, wherein.

R ₁ is selected from -(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 groups optionally substituted with one or more substituents selected from -(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;

X, Y and Z are independently selected from -O, -NH, -S, -N-(C ₁ -C ₃₀ )-alkyl or -(C ₁ -C ₃₀ )-aryl;

R ₂ is selected from -(C ₁ -C ₃₀ )-alkyl, =O, -OH, -SO ₂ , -SO, or -SOCH ₃ ; And

R ₄ is selected from -(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.

In another embodiment of the present invention there is provided a compound according to Formula IV:

or a pharmaceutically acceptable enantiomer, salt or solvate thereof.

The compound of Formula IV (also referred to as mJJ64A) was found to selectively activate Akt3. The IUPAC name for mJJ64A is 4-(m-{[p-(4-pyridylamino)phenylamino]carbonyl}phenylamino)-6-quinolinecarbonitrile. Since Akt3 modulates the suppressive function of natural Tregs and the polarization of induced Tregs, the compound of Formula IV and related compounds of Formulas I-III can be used to modulate immune responses.

In one embodiment, the present invention provides a method of enhancing an immunosuppressive response in a subject in need thereof, comprising administering to said subject a composition comprising a compound of Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof , which selectively activate Akt3 in an amount effective to enhance the immunosuppressive response in the subject.

For example, the present invention describes methods for increasing an immunosuppressive response, decreasing an immunostimulatory response, or a combination thereof in a subject in need thereof. The methods typically involve administering to a specified subject a composition comprising a compound of Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, that selectively activates the bioactivity of Akt3 in an amount effective to increase or stimulating an immunosuppressive response, reducing an immunostimulatory response, or a combination thereof in said subject.

In some embodiments of the present invention, the immunosuppressive response that is enhanced is selected from the group consisting of the immunosuppressive function of native Tregs (nTregs) and the promotion of conversion of typical T cells to induced Tregs (iTregs). The immunosuppressive function of nTregs may be the secretion of one or more anti-inflammatory cytokines. The anti-inflammatory cytokine(s) may be IL10, TGFβ, or a combination thereof.

In some embodiments of the present invention, said subject has an autoimmune disease. Therefore, this document also describes methods of treating autoimmune diseases by administering to a subject in need thereof an effective amount of a compound according to Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or a pharmaceutically acceptable salt thereof, that induces or increases the bioavailability or biological activity of Akt3.

Combination therapy including modulators of Akt3 biological activity and methods of its use are also described.

In one embodiment, the present invention provides a method of enhancing an immunosuppressive response in a subject in need thereof by administering to the subject a composition comprising an effective amount of a compound according to Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable compound thereof. salt that selectively activate Akt3 in an amount effective to enhance the immunosuppressive response in the subject. The subject may have an inflammatory disorder or disease, such as an autoimmune disease.

In another embodiment, the present invention provides a method of treating an inflammatory disorder in a subject in need thereof by administering a composition comprising a compound according to Formula I, Formula II, Formula III or Formula IV, or an enantiomer, polymorph or pharmaceutically acceptable salt thereof, which selectively activate Akt3 in an amount effective to enhance, induce or stimulate an immunosuppressive response in the specified subject.

In some embodiments of the present invention, the inflammatory disorder or disease is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome (ALPS), autoimmune thrombocytopenic purpura (ATP), Behçet's disease, bullous pemphigoid, cardiomyopathy, dermatitis associated with celiac sprue, chronic fatigue and immunodeficiency syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, cicatricial pemphigoid, cold agglutinin disease , syndrome of the cross, Crohn's disease, DEGO disease, dermatomyosite, dermatomyositis of juvenile, discoid lupus, essential mixed cryoglobulinemia, fibromiositis, Graves disease, Giyen -Barre disease, Hashimoto thyroeiditis, idiopathic fibrosis, idiopathic thromotopenic purple (I Tp), iga- nephropathy, insulin-dependent diabetes (type I), juvenile arthritis, Meniere's disease, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, obesity, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobus lineemia , primary biliary cirrhosis, psoriasis, Raynaud's phenomenon, Reiter's syndrome, rheumatic fever, sarcoidosis, scleroderma, Sjögren's syndrome, stiff muscle syndrome, Takayasu's arteritis, temporal arteritis/giant cell arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo and Wegener's granulomatosis.

In another embodiment, the present invention provides a method of treating an autoimmune disease by administering to a subject in need thereof a composition comprising a compound according to Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof that selectively inhibit Akt3 in an amount effective to stimulate or enhance an immunosuppressive response in the subject.

Common autoimmune diseases include, but are not limited to, rheumatoid arthritis, systemic lupus erythematosus, alopecia areata, autoimmune Addison's disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome (ALPS), autoimmune thrombocytopenic purpura (ATP), multiple sclerosis, Crohn's disease and myasthenia gravis.

In some embodiments of the present invention, the immunosuppressive response that is enhanced is selected from the group consisting of the immunosuppressive function of native Tregs (nTregs) and the induction of conversion of generic T cells to induced Tregs (iTregs). The immunosuppressive function of nTregs may include the secretion of one or more anti-inflammatory cytokines, such as IL-10, TGFβ, or a combination thereof.

In another embodiment, the present invention provides a method of treating a subject in need thereof by administering an effective amount of a composition comprising a compound according to Formula I, Formula II, Formula III or Formula IV, or an enantiomer, polymorph or pharmaceutically acceptable salt thereof in combination or alternating with a second immunosuppressive agent. Typical immunosuppressive agents include, but are not limited to, prednisone, budesonide, prednisolone, cyclosporine, tacrolimus, sirolimus, everolimus, azathioprine, leflunomide, mycophenolate, abatacept, adalimumab, anakinra, certolizumab, etanercept, golimumab, infliximab, ixekizumab, natalizumab , rituximab, secukinumab, tocilizumab, ustekinumab, vedolizumab, basiliximab, daclizumab, muromonab or combinations thereof.

In some embodiments of the present invention, a compound of Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, is administered in an amount effective to increase FoxP3 expression on immune cells, e.g., T cells, including , but not limited to, Treg cells such as iTreg and nTreg.

In other embodiments of the present invention, a compound of Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, is administered in an amount effective to enhance iTreg and nTreg proliferation.

Another embodiment of the present invention provides a pharmaceutical composition containing a compound according to Formula I, Formula II, Formula III or Formula IV or an enantiomer, polymorph or pharmaceutically acceptable salt and an excipient thereof. A compound of Formula I, Formula II, Formula III, Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof is generally present in an amount effective to enhance a suppressive immune response when administered to a subject in need thereof.

In another embodiment, the present invention provides a method of enhancing an immune suppressive response in a subject in need thereof by contacting immune cells ex vivo with a compound according to Formula I, Formula II, Formula III or Formula IV or an enantiomer, polymorph or pharmaceutical thereof an acceptable salt in an amount effective to increase FoxP3 expression on immune cells and administer the contacted immune cells to the subject. In one embodiment of the present invention, said immune cells are autologous immune cells. Said immune cells may include T cells, including, but not limited to, nTregs and iTregs.

In another embodiment, the present invention provides a method of inhibiting or reducing transplant rejection in a subject in need thereof by administering to the subject an effective amount of a compound according to Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or a pharmaceutically acceptable salt thereof to increasing FoxP3 expression on immune cells of said subject. In some embodiments of the present invention, increased expression of FoxP3 on immune cells of subjects induces, stimulates, or enhances a suppressive immune response in the subject.

In another embodiment, the present invention provides a method of treating graft-versus-host disease in a subject in need thereof by administering an effective amount of a compound according to Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or a pharmaceutically acceptable salt thereof to increase the expression of FoxP3 on immune cells of the specified subject.

In another embodiment, the present invention provides a method of treating a chronic infection in a subject in need thereof by administering an effective amount of a compound according to Formula I, Formula II, Formula III or Formula IV or an enantiomer, polymorph or pharmaceutically acceptable salt thereof to enhance FoxP3 expression on the immune cells of the specified subject.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

In FIG. 1 shows the structure of the compound of formula IV (mJJ64A).

Fig. 2A-2F are histograms of FACS-sorted iTregs treated with the indicated amounts of mJJ64A. Fig. 2G is a bar graph showing the percentage of CD4+ Foxp3+ T cells treated with the indicated amount of mJJ64A. Fig. 2H is a histogram of mean fluorescence intensity (MFI) (CD4+Foxp3 cells) treated with the indicated amount of mJJ64A.

Fig. 3A is a Western blot autoradiography showing Foxp3 expression upon iTreg induction and treatment with the indicated amount of mJJ64A. β-actin is the control. Fig. 3B is a repeat of the experiment in FIG. 3A.

Fig. 4A is a histogram representing the expression of three Akt isoforms (Akt1, Akt2 and Akt3) in Tconv (gray bar) and Treg (black bar) cells. Fig. 4B is a Western blot showing the expression of Akt1, Akt2 and Akt3 in Tconv and Treg cells. β-actin is a loading control. Fig. 4C is a Western blot showing pSer expression upon IP administration of Akt1, Akt2, or Akt3 in unstimulated or stimulated Tconv and nTreg cells. Fig. 4D is a histogram showing Treg suppressive activity in WT (gray bar) and Akt3 KO (black bar) mice. The X-axis represents the ratio of Tconv to Treg. The Y axis represents the normalized percentage of proliferation. Fig. 4E is a bar graph showing IL-10 levels (pg/ml) in Tregs from WT, Akt1 KO, Akt2 KO, or Akt3 KO mice. Fi. 4F is a bar graph showing TGFβ levels (pg/ml) in Tregs from WT, Akt1 KO, Akt2 KO, or Akt3 KO mice.

Fig. 5A is a line graph showing the percentage of survival of mice with RAG colitis treated with PBS (•), WT naïve T cells+PBS ( ), WT naïve T cells+WT nTreg ( ) or WT naïve T cells+Akt3 nTreg ( ). Fig. 5B is a line graph showing tumor volume (mm ³ ) over time (days) in EL4 (♦), PC61+EL4 (◊), PC61+EL4+WT Treg mice ( ) or PC61+EL4+Akt3 KO Treg ( ).

Fi. 6A is a Western blot showing the expression of Akt1, Akt2 and Akt3 in Th and iTreg cells. β-actin is a loading control. Fig. 6B is a histogram showing RNA expression of Akt1, Akt2 and Akt3 in Th (gray bar) and iTreg (black bar) cells. Fig. 6C is a Western blot showing expression of pSer in Th or Treg cells following IP administration of Akt1, Akt2 or Akt3. Fig. 6D is a histogram showing the percentage of FoxP3 cells⁺ in the CD4 T cell population in Th (gray bar) and iTreg cells (black bar) in Akt1 KO mice, Akt2 KO mice, and Akt3 KO mice. Figure 6E is a histogram showing the induction of FoxP3 in Tconv in response to TGFβ in Th (WT) + CV (solid line), iTreg (WT) + CV (dotted line), and iTreg (WT) + Akt3 shRNA (dashed line ).

Fig. 7A is a Western blot showing Akt3 and FoxP3 expression in control and Akt3 knockin Tregs. β-actin is used as a loading control. Fig. 7B is a histogram showing FoxP3 expression. Fig. 7C is a histogram showing MFI (CD4+FoxP3) in control and Akt3 KI Tregs. In FIG. 7D-I shows histograms representing IL2 and actin expression in control and Akt3 KI Tregs. In FIG. 7D and FIG. 7G demonstrates the expression of IL2 in controls and Akt3 KI, respectively. In FIG. 7E and FIG. 7H demonstrates the expression of actin in control and Akt3 KI, respectively. In FIG. 7F and 7I show overlay of IL2 and actin expression peaks for control and Akt3 KI, respectively.

Fig. 8 is a bar graph showing the effect of mJJ64A on the expression of Akt1 pS473, Akt2 pS474, Akt3 pS472 and Akt pan S473,474,472 in A2780 cells compared to control. The Y axis represents the integrated density value.

Fig. 9A is a Western blot showing the expression of FoxP3, pAkt3, Akt3, pAkt1, and Akt1 in activated Tconv cells induced by TGF-β and treated with various concentrations of mJJ64A. Fig. 9B is a histogram showing the expression of RORγt and FoxP3 in activated iTregs induced by TGF-β and treated with various concentrations of mJJ64A. Fig. 9C is a Western blot showing FoxP3 expression in activated iTregs induced by TGF-β and treated with various concentrations of mJJ64A.

In FIG. 10A shows histograms representing the proliferation of activated iTregs induced by TGF-β and treated with various concentrations of mJJ64A. Fig. 10B is a bar graph showing the percentage of proliferation of iTregs treated with various concentrations of mJJ64A. The X-axis represents the concentration of mJJ64A. The Y axis represents the percentage of proliferation. Fig. 10C is a histogram showing the percentage of live cells in iTregs treated with various concentrations of mJJ64A. The X-axis represents the treatment and the Y-axis represents the percentage of live cells. Fig. 10D is a set of histograms showing the proliferation of activated nTregs treated with various concentrations of mJJ64A. Fig. 10E is a histogram showing the percentage of proliferation of nTregs treated with various concentrations of mJJ64A. The X-axis represents the treatment and the Y-axis represents the percentage of proliferation. Fig. 10F is a histogram showing the percentage of live cells in nTregs treated with various concentrations of mJJ64A. The X-axis represents the treatment and the Y-axis represents the percentage of live cells. Fig. 10G is a set of histograms showing the proliferation of CD4 T cells treated with various concentrations of mJJ64A. Fig. 10H is a histogram showing the percentage of proliferation of CD4 T cells treated with different concentrations of mJJ64A. The X-axis represents the concentration of mJJ64A. The Y axis represents the percentage of proliferation. Fig. 10I is a histogram showing the percentage of live cells in CD4 T cells treated with various concentrations of mJJ64A. The X-axis represents the treatment and the Y-axis represents the percentage of live cells. Fig. 10J is a set of histograms showing the proliferation of CD8 T cells treated with various concentrations of mJJ64A. Fig. 10K is a histogram showing the percentage of proliferation of CD8 T cells treated with different concentrations of mJJ64A. The X-axis represents the concentration of mJJ64A. The Y axis represents the percentage of proliferation. Fig. 10L is a histogram showing the percentage of live cells in CD8 T cells treated with various concentrations of mJJ64A. The X-axis represents the treatment and the Y-axis represents the percentage of live cells.

Fig. 11A is a set of histograms demonstrating the suppressive function of mouse iTregs in untreated and mJJ64A-treated iTregs. The ratio of iTreg cells to Tconv cells was 0:1, 0.5:1, 1:1 and 2:1. Fig. 11B is a histogram showing the percentage of proliferation in untreated (black bar) and mJJ64A-treated (gray bar) cells at a Treg:Tconv ratio of 0.5:1. The X-axis represents the experimental group, and the Y-axis represents the percentage of proliferation. Fig. 11C is a histogram showing the percentage of proliferation in untreated (black bar) and mJJ64A-treated (gray bar) cells at a Treg:Tconv ratio of 1:1. The X-axis represents the experimental group, and the Y-axis represents the percentage of proliferation. Fig. 11D is a histogram showing the percentage of proliferation in untreated (black bar) and mJJ64A-treated (gray bar) cells at a Treg:Tconv ratio of 2:1. The X-axis represents the experimental group, and the Y-axis represents the percentage of proliferation.

Fig. 12A is a set of histograms demonstrating the suppressive function of untreated and mJJ64A-treated nTregs. The ratio of iTreg and Tconv cells was 0:1, 0.5:1, 1:1, 2:1 and 3:1. Fig. 12B is a histogram showing the percentage of proliferating cells in untreated and mJJ64A-treated nTregs in a mixture of nTregs and Tconvs (0.5:1). Fig. 12C is a histogram showing the percentage of proliferating cells in untreated and mJJ64A-treated nTregs in a 1:1 mixture of nTregs and Tconvs. Fig. 12D is a histogram showing the percentage of proliferating cells in untreated and mJJ64A-treated nTregs in a 2:1 mixture of nTregs and Tconvs. Fig. 12E is a histogram showing the percentage of proliferating cells in untreated and mJJ64A-treated nTregs in a 3:1 mixture of nTregs and Tconvs. Fig. 12F is a histogram showing the proliferation of nTregs treated with various concentrations of mJJ64A. Fig. 12G is a histogram showing the percentage of proliferation of nTregs treated with different concentrations of mJJ64A. The X-axis represents the treatment group and the Y-axis represents the percentage of proliferation. Fig. 12H is a histogram showing the percentage of live cells in nTregs treated with various concentrations of mJJ64A. The X-axis represents the treatment group and the Y-axis represents the percentage of proliferation.

Fig. 13A is a set of histograms showing FoxP3 and IL10 expression in nTreg cells treated with various concentrations of mJJ64A. Fig. 13B is a histogram showing the percentage of IL-10 ⁺ FoxP3 ⁺ cells in nTregs treated with various concentrations of mJJ64A.

Fig. 14A is an illustration showing an experimental method and treatment regimen for TC-1 tumor experiments. Fig. 14B is a line graph showing tumor volume (cm ³ ) over time (days) for non-treated ( ) and mJJ64A-(♦)-treated mice with TC1 tumor.

Fig. 15A is a histogram showing the number of CD8 cells⁺on 10⁶ live cells in tumors from untreated (black bar) and mJJ64A-treated (gray bar) mice. Fig. 15B is a histogram showing CD4 cell count⁺on 10⁶ live cells in tumors from untreated (black bar) and mJJ64A-treated (gray bar) mice. Fig. 15C is a histogram showing the number of FoxP3 cells⁺on 10⁶ CD4⁺cells in tumors from untreated (black bar) and mJJ64A-treated (gray bar) mice. Fig. 15D is a histogram showing the number of CD8 cells⁺, FoxP3^N.E.G. CD4⁺, and Treg by 10⁶live cells in the spleen of untreated (dark gray bar) or mJJ64A-treated (light gray bar) tumor-bearing mice. Fig. 15E is a histogram showing CD8 cell count⁺, FoxP3NEG CD4⁺and Treg by 10⁶live cells in the spleens of untreated (black column) or mJJ64A-treated (light gray column) tumor-free mice.

Fig. 16A is a schematic illustration of an experimental design of a colitis model. Fig. 16B is a line graph showing weight (g) over time (days post-injection) in colitis mice treated with control (•), iTreg ( ), mJJ64A+iTreg ( ) and mJJ64A ( ). Fig. 16C is a line graph showing normalized weight over time (days post-injection) in colitis mice treated with control (•), iTreg ( ), mJJ64A+iTreg ( ) and mJJ64A ( ). Fig. 16D is a line graph showing the percentage survival of mice with colitis not treated with any compound (•), iTreg (blue circle), mJJ64A+iTreg ( ) and mJJ64A (red circle). The X-axis represents time (days) and the Y-axis represents percentage survival. Fig. 16E-I are representative photographs of mice with colitis not receiving any compound (Figure 16E), receiving iTregs (Figure 16F), receiving JJa-treated iTregs (Figure 16G), not receiving any compound (Figure 16F) 16H) and receiving mJJ64A (Fig. 16I). The bottom image shows rectal prolapse in the groups not receiving any compound.

Fig. 17A is a photograph showing representative colons of untreated (UT), iTreg-treated, mJJ64A-treated iTreg, mJJ64A-treated, and wild-type (WT) mouse colons. In FIG. 17B is a histogram representing colon length and weight in healthy untreated (UT) and iTreg, mJJ64A, and mJJ64A+iTreg mice. The X-axis represents treatment group and the Y-axis represents colon weight/length (mg/mm). In FIG. 17C-J show representative histological sections of the colon: normal colon from WT mice (Figure 17C), Rag ^-/- colit- mice not treated with any compound (Figure 17D), normal colon from WT mice (Figure 17E ), Rag ^-/- colitis- mice not receiving any compound (Figure 17F)), Rag ^-/- colitis- mice receiving iTreg (Figure 17G), Rag ^-/- colitis- mice receiving mJJ64A ( 10 mg/kg) (Figure 17H), iTreg-treated Rag ^-/- colitis- mice (Figure 17I), and mJJ64A (10 mg/kg)-treated Rag ^-/- colitis- mice (Figure 17J).

Fig. 18A is a histogram showing the number of CD4 ⁺ T cells per 10 ⁶ live cells in the spleen of Rag ^-/- WT mice, UT mice, iTreg-treated mJJ64A mice, and mJJ64A-treated iTreg mice. The X-axis represents treatment group and the Y-axis represents the number of CD4 ⁺ cells per 10 ⁶ live cells. Fig. 18B is a histogram showing the percentage of FoxP3 ⁺ cells on CD4 ⁺ T cells in the spleen of Rag ^−/− WT mice, UT mice, iTreg-treated mJJ64A mice, and mJJ64A-treated iTreg mice. The X-axis represents treatment group and the Y-axis represents the number of FoxP3 ⁺ cells per number of CD4 ⁺ cells. Fig. 18C is a histogram showing the percentage of FoxP3 ⁻ cells per number of CD4 ⁺ T cells in the spleen of Rag ^−/− WT mice, UT mice, iTreg-treated mJJ64A mice, and mJJ64A-treated iTreg mice. The X-axis represents treatment group and the Y-axis represents the number of FoxP3 ⁻ cells per number of CD4 ⁺ cells. Fig. 18D is a histogram showing the number of CD4 ⁺ T cells per 10 ⁶ living cells in the lymph node of Rag ^-/- WT mice, UT mice, iTreg-treated mJJ64A mice, and mJJ64A-treated iTreg mice. The X-axis represents treatment group and the Y-axis represents the number of CD4 ⁺ cells per 10 ⁶ live cells. Fig. 18E is a histogram showing the percentage of FoxP3 ⁺ cells on CD4 ⁺ T cells in the lymph node of Rag ^−/− WT mice, UT mice, iTreg-treated mJJ64A mice, and mJJ64A-treated iTreg mice. The X-axis represents treatment group and the Y-axis represents the number of FoxP3 ⁺ cells per number of CD4 ⁺ cells. Fig. 18F is a histogram showing the percentage of FoxP3 ⁻ cells per number of CD4 ⁺ T cells in the lymph node of Rag ^−/− WT mice, UT mice, iTreg-treated mJJ64A mice, and mJJ64A-treated iTreg mice. The X-axis represents treatment group and the Y-axis represents the number of FoxP3 ⁺ cells per number of CD4 ⁺ cells.

Fig. 19A is a schematic illustration of the induction of an experimental model of autoimmune encephalomyelitis (EAE). Fig. 19B is a chart showing a grading criterion for scoring the severity of EAE. Fig. 19C is a line graph showing EAE scores over time (days after EAE induction) for control (•), iTreg (blue circle) and mJJ64A-10 ( ). The X-axis represents time (days) and the Y-axis represents the EAE score. Fig. 19D is a line graph showing the percentage of survival over time (days) of mice not treated with any compound ( ), iTreg-treated mice (•), and mJJ64A-10-treated mice ( ). The X-axis represents time (days) and the Y-axis represents percentage survival. Fig. 19E is a line graph showing the EAE score over time (days after EAE induction) for EAE mice treated with control (•), iTreg (♦), mJJ64A-3 (blue circle), mJJ64A-6 ( ) and mJJ64A-10 ( ). The x-axis represents time (days after EAE induction) and the y-axis represents the EAE score. Fig. 19F is a line graph showing the percentage of survival over time (days) of EAE mice not treated with any compound ( ), iTreg-treated mice (blue circle), mJJ64A-3 ( ), mJJ64A-6 ( ) and mJJ64A-10 (♦). The X-axis represents time (days) and the Y-axis represents percentage survival.

Fig. 20A-C are histograms showing the percentage of FoxP3 ⁺ cells per number of CD4 ⁺ T cells in the spleen (Fig. 20A), blood (Fig. 20B) and brain (Fig. 20C) of EAE UT mice, EAE mice treated iTreg, mJJ64A-3, mJJ64A-6 and mJJ64A-10. The X-axis represents treatment group and the Y-axis represents the number of FoxP3 ⁺ cells per number of CD4 ⁺ cells. Fig. 20D-F are histograms showing the percentage of FoxP3 ^- cells per number of CD4 ⁺ T cells in the spleen (Fig. 20D), blood (Fig. 20E) and brain (Fig. 20F) of EAE UT mice, EAE mice treated iTreg, mJJ64A-3, mJJ64A-6 and mJJ64A-10. The X-axis represents treatment group and the Y-axis represents the number of FoxP3 ⁻ cells per number of CD4 ⁺ cells. Fig. 20G-I are histograms showing the percentage of ROR ⁺ cells per number of CD4 ⁺ T cells in the spleen (Fig. 20G), blood (Fig. 20H), and brain (Fig. 20I) of EAE UT mice, EAE mice, treated with iTreg, mJJ64A-3, mJJ64A-6 and mJJ64A-10. The X-axis represents treatment group and the Y-axis represents the number of ROR ⁺ cells per number of CD4 ⁺ cells.

Fig. 21A is a bar graph showing the percentage of live human iTregs in cells treated with various concentrations of mJJ64A. The X-axis represents the treatment group and the Y-axis represents the percentage of live cells. Fig. 21B is a bar graph showing the percentage of FoxP3 ⁺ CD4 ⁺ in human iTregs treated with various concentrations of mJJ64A. The X-axis represents treatment group and the Y-axis represents the percentage of FoxP3 ⁺ CD4 ⁺ cells.

DETAILED DESCRIPTION OF THIS INVENTION

I. Definitions

The term “promote expression” means to influence expression, eg, to induce expression or activity, or to induce increased/higher expression or activity compared to normal healthy controls.

The terms “immune-activating response,” “activating immune response,” and “immunostimulatory response” refer to a response that initiates, induces, enhances, or enhances the activation or effectiveness of the innate or adaptive immune system. Such immune responses include, for example, the development of a beneficial humoral (antibody-mediated) and/or cellular (antigen-specific T cell-mediated or their secretion products) response directed against the peptide in the recipient patient. Such a response may be an active response induced by administration of an immunogen, or a passive response induced by administration of an antibody or primed T cells. The cellular immune response is caused by the presentation of polypeptide epitopes in association with MHC class I or class II molecules to activate antigen-specific CD4 ⁺ T helper cells and/or CD8 ⁺ cytotoxic T cells. The response may also include activation of monocytes, macrophages, NK cells, basophils, dendritic cells, astrocytes, microglia cells, eosinophils, activation or recruitment of neutrophils, or other components of the innate immune system. The presence of a cellular immunological response can be determined using proliferation (CD4 ⁺ T cells) or CTL (cytotoxic T lymphocytes) assays. The relative contribution 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 an immunized syngeneic animal and measuring the protective or therapeutic effect in a second subject.

The terms "suppressive immune response" and "immunosuppressive response" refer to a response that reduces or prevents the activation or effectiveness of the innate or adaptive immune system.

As used herein, the term “immune tolerance” refers to any mechanism by which a potentially harmful immune response is prevented, suppressed, or biased towards a non-harmful immune response (Bach et al., N. Eng. J. Med. , 347:911-920 ( 2002)).

In this context, the term "tolerogenic vaccine" generally refers to an antigen-specific drug used to attenuate autoreactive T and/or B cell responses while leaving overall immune function unchanged.

An "immunogenic agent" or "immunogen" is capable of causing an immunological response against itself when administered to a mammal, optionally in combination with an adjuvant.

The term "immune cell" refers to cells of the innate and adaptive immune system, including neutrophils, eosinophils, basophils, monocytes, macrophages, dendritic cells, lymphocytes, including B cells, T cells and natural killer cells.

As used herein, "typical T cells" are T lymphocytes that express the α-T cell receptor (TCR) as well as the CD4 or CD8 co-receptor. Typical T cells are present 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).

As used herein, “atypical T cells” are lymphocytes that express the γδ TCR and may typically be found in an epithelial environment such as the skin, gastrointestinal tract, or genitourinary tract. Another subset of atypical T cells is the invariant natural killer T (NKT) cell, which has the phenotypic and functional capabilities of a typical T cell, as well as the features of natural killer cells (eg, cytolytic activity). See Roberts and Girardi, “Conventional and Unconventional T Cells,” Clinical and Basic Immunodermatology, pp. 85-104, (Gaspari and Tyring (ed.)), Springer London (2008).

As used herein, the term "Treg" refers to regulatory T cells or cells. Regulatory T cells are a subpopulation of T cells that modulate the immune system, maintain tolerance to self-antigens, eliminate autoimmune diseases, and otherwise suppress immunostimulatory or activating responses of other cells. Regulatory T cells come in many forms, the most studied of which are those that express CD4, CD25, and Foxp3.

As used herein, the term "natural Treg" or "nTreg" refers to regulatory T cells or cells that develop in the thymus.

As used herein, the term “induced Treg” or “iTreg” refers to regulatory T cells or cells that develop from mature conventional CD4+ T cells outside the thymus.

"Biological activity" of Akt3 refers to the biological function of the Akt3 polypeptide. Bioactivity can be increased or decreased by increasing or decreasing the activity of baseline levels of the polypeptide, increasing or decreasing the avidity of baseline levels of the polypeptide, the amount of polypeptide, the ratio of Akt3 relative to one or more other Akt isoforms (e.g., Akt1 or Akt2) of the polypeptide, increasing or decreasing polypeptide expression levels (including by increasing or decreasing Akt3 mRNA expression) or combinations thereof. For example, a bioavailable Akt3 polypeptide is a polypeptide that has kinase activity and can bind to and phosphorylate an Akt3 substrate. An Akt3 polypeptide that is not bioavailable includes an Akt3 polypeptide that is mislocalized or is unable to bind and phosphorylate Akt substrates.

As used herein, the phrase that a molecule “specifically binds” or “exhibits specific binding” to a target refers to a binding reaction that is determinative of the molecule's presence in the presence of a heterogeneous population of other biologicals.

Under certain immunoassay conditions, the molecule binds preferentially to a specific target and does not bind in significant amounts to other biological substances present in the sample. Specific binding of an antibody to a target under such conditions requires that the antibody be selected for its target specificity. Various immunoassay formats can be used to select antibodies that are specifically immunoreactive with a particular protein. For example, ELISA is commonly used to select monoclonal antibodies that are specifically immunoreactive with a protein. See, for example, Harlow and Lane (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Publications, New York, for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity.

The terms "oligonucleotide" and "polynucleotide" generally refer to any polyribonucleotide or polydeoxribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. Thus, for example, the term "polynucleotides" as used herein refers to, inter alia, single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double-stranded RNA, and RNA that is a mixture of single- and double-stranded regions, hybrid molecules containing DNA and RNA, which can be single-stranded or, more typically, double-stranded or a mixture of single-stranded and double-stranded regions. The term "nucleic acid" or "nucleic acid sequence" also includes a polynucleotide as defined above.

In addition, the term "polynucleotide" as used herein refers to tristranded regions containing RNA or DNA, or both RNA and DNA. The chains in such regions may be from the same molecule or from different molecules. The regions may include all of one or more molecules, but typically only include a region of some of the molecules. One of the molecules in the trihelix region is often an oligonucleotide.

As used herein, the term "polynucleotide" includes DNA or RNA, as described above, that contains one or more modified bases. Thus, DNA or RNA with scaffolds modified for stability or other reasons are “polynucleotides,” which term is intended for use herein. In addition, DNA or RNA containing atypical bases, such as inosine, or modified bases, such as tritylated bases, to give only two examples, are polynucleotides and are encompassed by this term as defined herein.

As used herein, the term "polypeptide" refers to a chain of amino acids of any length regardless of modification (eg, phosphorylation or glycosylation). The term polypeptide includes proteins and their fragments. The polypeptides may be "exogenous", meaning that they are "heterologous", that is, foreign to the host cell being used, for example, a human polypeptide produced by a bacterial cell. Polypeptides are described herein as sequences of amino acid residues. These sequences are written from left to right in the direction from the amino to the carboxy terminus. According to standard nomenclature, sequences of amino acid residues are designated by a three-letter or one-letter code 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).

"Variant" refers to a polypeptide or polynucleotide that differs from, but retains essential properties of, the reference polypeptide or polynucleotide. A typical polypeptide variant differs in amino acid sequence from another reference polypeptide. Typically the differences are limited such that the sequences of the reference polypeptide and the variant are generally very similar and identical in many areas. The variant and the reference polypeptide may differ in amino acid sequence by one or more modifications (eg, substitutions, additions and/or deletions). The substituted or inserted amino acid residue may or may not be genetically encoded. A polypeptide variant may occur naturally, such as an allelic variant, or it may be a variant that is not known to occur in nature.

Modifications and changes can be made to the structure of the polypeptides of this invention and still produce a molecule having characteristics similar to the polypeptide (eg, a conservative amino acid substitution). For example, some amino acids can be replaced by other amino acids in the sequence without noticeable loss of activity. Since it is the interactivity and nature of a polypeptide that determines the biological functional activity of that polypeptide, certain amino acid sequence substitutions can be made in the polypeptide sequence and still result in a polypeptide with similar properties.

When making such changes, the hydrophobicity index of amino acids can be taken into account. Those skilled in the art generally understand the importance of the hydrophobicity index of amino acids in imparting interactive biological function to a polypeptide. It is known that certain amino acids can be replaced by other amino acids having a similar index or hydrophobicity index, but this results in a polypeptide with similar biological activity. Each amino acid is assigned a hydrophobicity index based on its hydrophobicity and charge characteristics. These indices are as follows: 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).

The nature of the relative hydrophobicity of an amino acid is thought to determine the secondary structure of the resulting polypeptide, which in turn determines the interaction of the polypeptide with other molecules such as enzymes, substrates, receptors, antibodies, antigens, and cofactors. It is known in the art that an amino acid can be replaced with another amino acid having a similar hydrophobicity index and still produce a functionally equivalent polypeptide. With such changes, substitution of amino acids whose hydrophobic indices are within ± 2 is preferred, those hydrophobic indices which are within ± 1 are particularly preferred, and those hydrophobic indices which are within ± 0.5 are even more particularly preferred.

Substitution of similar amino acids can also be made on the basis of hydrophilicity, in particular when the biologically functional equivalent polypeptide or peptide thus created is intended for use in immunological embodiments. The amino acid residues were assigned the following hydrophilicity values: 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); tryptophan (- 3.4). One skilled in the art will appreciate that an amino acid can be replaced by another amino acid having a similar hydrophilicity value and still produce a biologically equivalent and, in particular, an immunologically equivalent polypeptide. With such changes, substitution of amino acids whose hydrophilicity indices are within ± 2 is preferred, those hydrophilicity indices which are within ± 1 are particularly preferred, and those hydrophilicity indices which are within ± 0.5 are even more particularly preferred.

As stated above, amino acid substitutions are generally based on the relative similarity of the side chain amino acid substituents, such as their hydrophobicity, hydrophilicity, charge, size, and the like. Exemplary substitutions that take into account the various above-mentioned characteristics 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). Embodiments of the present invention thus provide functional or biological equivalents of the polypeptide as set forth above. In particular, embodiments of polypeptides may include variants having about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity with the polypeptide of interest .

The term "percentage (%) sequence identity" is defined as the percentage of nucleotides or amino acids in a candidate sequence that are identical to the nucleotides or amino acids in a reference nucleic acid sequence after sequence alignment and the introduction of gaps, if necessary, to achieve maximum percent sequence identity. Alignment for the purpose of determining percent amino acid sequence identity can be accomplished in a variety of ways known to those skilled in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2, or Megalign (DNASTAR) software. Appropriate parameters for measuring alignment, including any algorithms necessary to achieve maximum alignment over the entire length of the sequences being compared, can be determined using known methods.

For the purposes of this invention, the % identity of a given nucleotide or amino acid sequence C with, compared to or against a given nucleic acid sequence D (which may alternatively be stated as a given amino acid sequence C that has or contains a certain % sequence identity with, to compared with or against a given sequence D) is calculated as follows:

100x W/Z ratio

where W is the number of nucleotides or amino acids that were judged to be identical matches when aligning C and D using a sequence alignment program, and where Z is the total number of nucleotides or amino acids in D. One skilled in the art will appreciate that if the length of the amino acid sequence C is not equal to the length of sequence D, the % identity of the sequence C with respect to D will not be equal to the % identity of the sequence D with respect to C.

The term "carrier" refers to an organic or inorganic ingredient, natural or synthetic, with which the active ingredient is combined to facilitate administration.

The term "pharmaceutically acceptable" means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredients.

The term "pharmaceutically acceptable carrier" means one or more compatible solid or liquid excipients, diluents or encapsulating substances that are suitable for administration to a human or other vertebrate animal.

The term “effective amount” or “therapeutically effective amount” means a dose sufficient to provide treatment for the disorder, disease or condition being treated, or to otherwise provide a desired pharmacological and/or physiological effect. The exact dosage will vary depending on many factors, such as subject-specific variables (eg, age, immune system status, etc.), disease, and treatment being administered.

The terms "individual", "subject" and "patient" are used interchangeably herein and refer to a mammal, including, but not limited to, humans, rodents such as mice and rats, and other laboratory animals.

II. Compositions for activating Akt 3

The present invention provides compositions and methods of their use for selective activation of Akt3.

In one embodiment of the present invention there is provided a compound of formula I:

or a pharmaceutically acceptable enantiomer, salt or solvate thereof, wherein:

rings A, B and C are independently six-membered aryl or N-containing heteroaryl mono- 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.

R ₁ is selected from -(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 groups optionally substituted with one or more substituents selected from -(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;

X, Y and Z are independently selected from =O, -NH, -S, -N-(C ₁ -C ₃₀ )-alkyl or -(C ₁ -C ₃₀ )-aryl;

R ₂ is selected from -(C ₁ -C ₃₀ )-alkyl, =O, -OH, -SO ₂ , -SO, or -SOCH ₃ ; And

R ₃ is selected from -(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 groups, optionally substituted by one or more substituents selected from -(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.

In another embodiment of the present invention there is provided a compound of formula II:

or a pharmaceutically acceptable enantiomer, salt or solvate thereof, wherein:

R ₁ is selected from -(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 groups optionally substituted with one or more substituents selected from -(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;

X, Y and Z are independently selected from -O, -NH, -S, -N-(C ₁ -C ₃₀ )-alkyl or -(C ₁ -C ₃₀ )-aryl;

R ₂ is selected from -(C ₁ -C ₃₀ )-alkyl, -O, -OH, -SO ₂ , -SO, or -SOCH ₃ ; And

R ₃ is selected from -(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 groups, optionally substituted by one or more substituents selected from -(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.

In another embodiment of the present invention there is provided a compound of formula III:

or a pharmaceutically acceptable enantiomer, salt or solvate thereof, wherein:

R ₁ is selected from -(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 groups optionally substituted with one or more substituents selected from -(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;

X, Y and Z are independently selected from -O, -NH, -S, -N-(C ₁ -C ₃₀ )-alkyl or -(C ₁ -C ₃₀ )-aryl;

R ₂ is selected from -(C ₁ -C ₃₀ )-alkyl, -O, -OH, -SO ₂ , -SO, or -SOCH ₃ ; And

R ₄ is selected from -(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.

In another embodiment of the present invention there is provided a compound of formula IV:

or a pharmaceutically acceptable enantiomer, salt or solvate thereof.

The compound of formula IV, also referred to as mJJ64A, and enantiomers, polymorphs, pharmaceutically acceptable salts and derivatives thereof can be used to induce, stimulate or increase the biological activity of Akt3 in immune cells.

In some embodiments of the present invention, the Atk3 activator is a derivative of Formula I, Formula II, Formula III, or Formula IV. As used herein, the term "derivative" or "derivatized" includes one or more chemical modifications of Formula I, Formula II, Formula III or Formula IV or an enantiomer, polymorph or pharmaceutically acceptable salt thereof. That is, a “derivative” may be a functional equivalent of Formula I, Formula II, Formula III, or Formula IV that is capable of producing improved pharmacological functional activity and/or behavioral response in a given subject. An illustration of such chemical modifications may be the replacement of hydrogen with a halogen group, an alkyl group, an acyl group, or an amino group.

Chemical modification of Formula I, Formula II, Formula III, or Formula IV, an enantiomer, a polymorph, or a pharmaceutically acceptable salt thereof can either increase or decrease the hydrogen bonding interaction, charge interaction, hydrophobic interaction, van der Waals interaction, or dipole interaction between the specified compound and its target.

In some embodiments of the present invention, a compound of Formula I, Formula II, Formula III, or Formula IV may serve as a model (e.g., template) for the development of other derivative compounds that are functionally equivalent to the compound and that are capable of inducing improved pharmacological functional activity and/or or effect and/or behavioral response in a given subject.

The compound of formula I, formula II, formula III or formula IV may be a racemic compound and/or optically active isomers thereof. Because of this, some of the compounds may have asymmetric carbon atoms and therefore may exist either as racemic mixtures or as individual optical isomers (enantiomers). Compounds described herein that contain a chiral center include all possible stereoisomers of the compound, including compositions comprising a racemic mixture of two enantiomers, as well as compositions comprising each enantiomer alone, substantially free of the other enantiomer. Thus, for example, contemplated herein is a composition comprising the S-enantiomer of a compound substantially free of the R-enantiomer, or an R-enantiomer substantially free of the S-enantiomer. If said compound includes more than one chiral center, the scope of this invention also includes compositions including mixtures with varying proportions between diastereomers, as well as compositions including one or more diastereomers substantially free of one or more other diastereomers. By "substantially free" is meant that the composition includes less than 25%, 15%, 10%, 8%, 5%, 3%, or less than 1% of the minor enantiomer or diastereomer(s).

Akt3, also called RAC-gamma-serine/threonine protein kinase, is an enzyme encoded by the Akt3 gene in humans. Akt kinases are known to be regulators of cellular signaling in response to insulin and growth factors and are associated with a wide range of biological processes, including proliferation, differentiation, apoptosis, cell tumorigenesis, as well as glycogen synthesis and glucose uptake. Akt3 has been demonstrated to be stimulated by platelet-derived growth factor (PDGF), insulin, and insulin-like growth factor 1 (IGF1).

Akt3 kinase activity mediates serine and/or threonine phosphorylation of a number of downstream substrates. Nucleic acid sequences for Akt3 are known in the art. For example, see GenBank accession number AF124141.1: Homo sapiens protein kinase B gamma mRNA , complete cds, which are specifically incorporated by reference in their entirety and provide the nucleic acid sequence:

AGGGGAGTCATCATGAGCGATGTTACCATTGTGAAGGAAGGTTGGGTTCAGAAGAGGGGA

GAATATATAAAAAACTGGAGGCCAAGATACTTCCTTTGAAGACAGAATGGCTCATTCATA

GGATATAAAAGAGAAACCTCAAGATGTGGATTTACCTTATCCCCTCAACAACTTTTCAGTG

GCAAAATGCCAGTTAATGAAAACAGAACGACCAAAGCCAAACACATTTATAATCAGATGT

CTCCAGTGGACTACTGTTATAGAGAGAACATTTCATGTAGATACTCCAGAGGAAAGGGAA

GAATGGACAGAAGCTATCCAGGCTGTAGCAGACAGACTGCAGAGGCAAGAAGAGGAGAGAATGAATTGTAGTCCAACTTCACAAATTGATAATATAGGAGAGGAAGAGATGGATGCCTCT

ACAACCCATCATAAAAGAAAGACAATGAATGATTTTGACTATTTGAAACTACTAGGTAAA

GGCACTTTTGGGAAAGTTATTTTGGTTCGAGAGAAGGCAAGTGGAAAATACTATGCTATG

AAGATTCTGAAGAAAGAAGTCATTATTGCAAAGGATGAAGTGGCACACACTCTAACTGAA

AGCAGAGTATTAAAGAACACTAGACATCCCTTTTTAACATCCTTGAAATATTCCTTCCAG

ACAAAAGACCGTTTGTGTTTTGTGATGGAATATGTTAATGGGGGCGAGCTGTTTTTCCAT

TTGTCGAGAGAGCGGTGTTCTCTGAGGACCGCACACGTTTCTATGGTGCAGAAATTGTC

TTCTGCCTTGGACTATCTACATTCCGGAAAGATTGTGTACCGTGATCTCAAGTTGGAGAAT

CTAATGCTGGACAAAGATGGCCACATAAAAATTACAGATTTTGGACTTTGCAAAGAAGGG

ATCACAGAATGCAGCCACCATGAAGACATTCTGTGGCACTCCAGAATATCTGGCACCAGAG

GTGTTAGAAGATAATGACTATGGCCGAGCAGTAGACTGGTGGGGCCTAGGGGTTGTCATG

TATGAAATGATGTGTGGGAGGTTACCTTTCTACAACCAGGACCATGAGAAACTTTTTGAA

TTAATATTAATGGAAGACATTAAATTTTCCTCGAACACTCTCTTCAGATGCAAAATCATTG

CTTTCAGGGCTCTTGATAAAGGATCCAAATAAAACGCCTTGGTGGAGGACCAGATGATGCA

AAAGAAATTATGAGACACAGTTTCTTCTCTGGAGTAAACTGGCAAGATGTATATGATAAA

AAGCTTGTACCTCCTTTTAAACCCTCAAGTAACATCTGAGACAGATACTAGATATTTTGAT

GAAGAATTTTACAGCTCAGACTATTACAATAACACCACCTGAAAAATATGATGAGGATGGT

ATGGACTGCATGGACAATGAGAGGCGGCCGCATTTCCCTCAATTTTCCTACTCTGCAAGT

GGACGAGAATAAGTCTCTTTCATTCTGCTACTTCACTGTCATCTTCAATTTATTACTGAA

AATGATTCCTGGACATCACCAGTCCTAGCTCTTACACATAGCAGGGGCACCTTCCGACAT

CCCAGACAGCCAAGGGTCCTCACCCCTCGCCACCTTCACCCTCATGAAAACACACATA

CACGCAAATACACTCCAGTTTTTGTTTTTGCATGAAATTGTATCTCAGTCTAAGGTCTCA

TGCTGTTGCTGCTACTGTCTTACTATTA

(SEQ ID NO: 1)

Amino acid sequences are also known in the art. See eg accession number UniProtKB/Swiss-Prot. Q9Y243 (Akt3_HUMAN), which is specifically incorporated by reference in its entirety and provides the amino acid sequence:

MSDVTIVKEGWVQKRGEYIKNWRPRYFLLKTDGSFIGYKEKPQDVDLPYPLNNFSVAKCQ

LMKTERPKPNTFIIRCLQWTTVIERTFHVDTPEEREEWTEAIQAVADRLQRQEEERMNCS

PTSQIDNIGEEEMDASTTHHKRKTMNDFDYLKLLGKGTFGKVILVREKASGKYYAMKILK

KEVIIAKDEVAHTLTESRVLKNTRHPFLTSLKYSFQTKDRLCFVMEYVNGGELFFHLSRE

RVFSEDRTRFYGAEIVSALDYLHSGKIVYRDLKLENLMLDKDGHIKITDFGLCKEGITDA

ATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYEMMCGRLPFYNQDHEKLFELILM

EDIKFPRTLSSDAKSLLSGLLIKDPNKRLGGGPDDAKEIMRHSFFSGVNWQDVYDKKLVP

PFKPQVTSETDTRYFDEEFTAQTITITPPEKYDEDGMDCMDNERRPHFPQFSYSASGRE

(SEQ ID NO: 2).

The domain structure of Akt3 is discussed in Romano, Scientifica , Volume 2013 (2013), Article ID 317186, 12 pages, and includes an N-terminal pleckstrin homology (PH) domain followed by a kinase catalytic domain (KD) and a C- terminal regulatory hydrophobic region. 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 three Akt isoforms. The PH domain binds lipid substrates such as phosphatidylinositol (3,4) diphosphate (PIP2) and phosphatidylinositol (3,4,5) triphosphate (PIP3). The ATP binding site is located approximately in the middle of the catalytic domain of the kinase, which has a significant degree of homology with other components of the AGC kinase family, such as p70 S6 kinase (S6K) and p90 ribosomal S6 kinase (RSK), protein kinase A (PKA), and protein kinase B ( PKB). The hydrophobic regulatory moiety is a typical feature of the AGC kinase family. With reference to SEQ ID NO: 2, Akt 3 is believed to generally have the following molecule processing and domain structure described below.

Molecule Processing:

The initiator methionine SEQ ID NO: 2 is disposable for Akt3 function. Therefore, in some embodiments of the present invention, the compound directly or indirectly inhibits the expression or bioavailability of Akt3 having the amino acid sequence

SDVTIVKEGWVQKRGEYIKNWRPRYFLLKTDGSFIGYKEKPQDVDLPYPLNNFSVAKCQ

LMKTERPKPNTFIIRCLQWTTVIERTFHVDTPEEREEWTEAIQAVADRLQRQEEERMNCS

PTSQIDNIGEEEMDASTTHHKRKTMNDFDYLKLLGKGTFGKVILVREKASGKYYAMKILK

KEVIIAKDEVAHTLTESRVLKNTRHPFLTSLKYSFQTKDRLCFVMEYVNGGELFFHLSRE

RVFSEDRTRFYGAEIVSALDYLHSGKIVYRDLKLENLMLDKDGHIKITDFGLCKEGITDA

ATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYEMMCGRLPFYNQDHEKLFELILM

EDIKFPRTLSSDAKSLLSGLLIKDPNKRLGGGPDDAKEIMRHSFFSGVNWQDVYDKKLVP

PFKPQVTSETDTRYFDEEFTAQTITITPPEKYDEDGMDCMDNERRPHFPQFSYSASGRE

(SEQ ID NO: 3).

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

A. Compositions

In another embodiment, the present invention provides formulations and pharmaceutical compositions comprising one or more compounds of Formulas I, II, III, IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof. Typically, dose levels for the compounds described herein are from about 0.0001 mg/kg body weight to about 1000 mg/kg, more preferably from 0.001 to 500 mg/kg, more preferably from 0.01 to 50 mg /kg body weight per day for administration to mammals.

1. Delivery medium

The compounds of formulas I, II, III and IV can be administered to a subject, preferably a human, and are introduced into the cells of the subject with or without the aid of a delivery vehicle. Suitable carriers for delivering the described active agents are known in the art and can be selected according to the particular active agent. In some embodiments of this invention, the compound is incorporated into or encapsulated into a nanoparticle, microparticle, micelle, synthetic lipoprotein particle, or carbon nanotube. For example, these compositions can be included in a carrier, such as polymer microparticles, which provide controlled release of the active agent(s). In some embodiments of the present invention, the release of the drug(s) is controlled through the diffusion of the active agent(s) from the microparticles and/or the degradation of the polymer particles by hydrolysis and/or enzymatic degradation. Suitable polymers include ethylcellulose and other natural or synthetic cellulose derivatives. Polymers that dissolve slowly and form a gel in an aqueous environment, such as hydroxypropyl methylcellulose or polyethylene oxide, may also be suitable as materials for drug-containing microparticles. Other polymers include, but are not limited to, polyanhydrides, poly(ester anhydrides), polyhydroxy acids such as polylactide (PLA), polyglycolide (PGA), poly(lactide-co-glycolide) (PLGA), poly-3-hydroxybutyrate (PHB) ) and its copolymers, poly-4-hydroxybutyrate (P4HB) and its copolymers, polycaprolactone and its copolymers and combinations thereof.

In some embodiments of this invention, compounds of Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, and the second therapeutic agent are included in the same particles and formulated to be released at different times and/or or at different periods of time. For example, in some embodiments of the present invention, one of the agents is completely released from the particles before the release of the second agent begins. In other embodiments of this invention, the release of the first agent occurs followed by the release of the second agent before the entire amount of the first agent is released. In other embodiments of this invention, both agents are released at the same time for the same period of time or for different periods of time.

Compounds according to Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, may be included in a delivery vehicle derived from materials that are insoluble in aqueous solution or slowly soluble in aqueous solution but capable of degrade in the gastrointestinal tract through enzymatic degradation, surfactant action of bile acids, and/or mechanical erosion. As used herein, the term "slowly soluble in water" refers to materials that do not dissolve in water within 30 minutes. Preferred examples include fats, fatty substances, waxes, waxy substances and mixtures thereof. Suitable fats and fatty substances include fatty alcohols (such as lauryl, myristyl stearyl, cetyl or cetostearyl alcohol), fatty acids and derivatives including, but not limited to, fatty acid esters, fatty acid glycerides (mono-, di- and tri- glycerides) and hydrogenated fats. Specific examples include, but are not limited to, hydrogenated vegetable oil, hydrogenated cottonseed oil, hydrogenated castor oil, hydrogenated oils available under the trade name Sterotex®, stearic acid, cocoa butter, and stearyl alcohol. Suitable waxes and wax-like materials include natural or synthetic waxes, hydrocarbons and conventional waxes.

Specific examples of waxes include beeswax, glycowax, castor wax, carnauba wax, paraffins and candelilla wax. As used herein, the term "waxy material" is defined as any material that is generally solid at room temperature and has a melting point of about 30 to 300°C. The release point and/or release period may vary, as discussed above.

2. Pharmaceutical compositions

The present invention provides pharmaceutical compositions comprising compounds according to Formula I, Formula II, Formula III or Formula IV with or without a delivery vehicle. Pharmaceutical compositions may be formulated for administration via parenteral (intramuscular, intraperitoneal, intravenous (IV) or subcutaneous administration), enteral, transmucosal (nasal, vaginal, rectal or sublingual) or transdermal (either passive or by iontophoresis or electroporation ) routes of administration or through the use of biodegradable inserts and can be formulated in dosage forms suitable for each route of administration.

In certain embodiments of this invention, these compositions are administered topically, for example, by injection directly into the site to be treated (eg, a tumor). In some embodiments of the present invention, the compositions are injected or otherwise administered directly into the vasculature of vascular tissue proximate or adjacent to the intended treatment site (eg, adjacent to a tumor). Typically, local administration will result in increased local concentrations of the composition that are greater than what can be achieved with systemic administration.

A. Formulations for parenteral administration

Compounds according to formula I, formula II, formula III or formula IV or an enantiomer, polymorph or pharmaceutically acceptable salt thereof and pharmaceutical compositions thereof can be administered in aqueous solution by parenteral injection. Said composition may also be in the form of a suspension or emulsion. In general, pharmaceutical compositions are provided with effective amounts of the active agent(s) and optionally include pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants and/or carriers. Such compositions include diluents, sterile water, buffered saline with varying buffer content (eg, Tris-HCl, acetate, phosphate), pH, and varying ionic strength; and, optionally, additives such as detergents and solubilizing agents (e.g., TWIN® 20, TWIN® 80, also called polysorbate 20 or 80), antioxidants (e.g., ascorbic acid, sodium metabisulfite), and preservatives (e.g., thimerosol, benzyl alcohol ) and fillers (eg lactose, mannitol). Examples of non-aqueous solvents or carriers include propylene glycol, polyethylene glycol, vegetable oils such as olive oil and corn oil, gelatin, and injectable organic esters such as ethyl oleate. These formulations may be lyophilized and re-dissolved/re-suspended immediately prior to use. The composition can be sterilized, for example, by filtration through a bacteria-retaining filter, inclusion of sterilizing agents in the compositions, irradiation of the compositions, or heating of the compositions.

b. Formulations for enteral administration

Compounds according to Formula I, Formula II, Formula III or Formula IV, or an enantiomer, polymorph or pharmaceutically acceptable salt thereof, can be formulated for enteral administration. Suitable oral dosage forms of the compounds of formula I, formula II, formula III or formula IV or an enantiomer, polymorph or pharmaceutically acceptable salt thereof include tablets, capsules, solutions, suspensions, syrups and troches. Tablets can be prepared using compression or molding techniques well known in the art. Gelatin or non-gelatin capsules can be formulated as hard or soft shell capsules that can encapsulate liquid, solid and semi-solid fillers using methods well known in the art.

The compositions can be prepared using a pharmaceutically acceptable carrier. As used herein, the term "carrier" generally includes, but is not limited to, diluents, preservatives, binding agents, lubricants, disintegrants, swelling agents, fillers, stabilizers, and combinations thereof.

The carrier also includes all components of the coating composition, which may include plasticizers, pigments, dyes, stabilizers and glidants. Sustained release dosage formulations can be prepared as described in the well-known literature. These sources provide information on carriers, materials, equipment and methods for preparing tablets and capsules and sustained-release dosage forms of tablets, capsules and granules.

Examples of suitable coating materials include, but are not limited to, cellulose polymers such as cellulose acetate phthalate, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate and hydroxypropylmethylcellulose acetate succinate; polyvinyl acetate phthalate, polymers and copolymers of acrylic acid and methacrylic resins, which are commercially available under the trade name Eudragit® (Roth Pharma, Westerstadt, Germany), zein, shellac and polysaccharides.

In addition, the coating material may contain conventional carriers such as plasticizers, pigments, dyes, glidants, stabilizers, blowing agents and surfactants.

Optional pharmaceutically acceptable excipients include, but are not limited to, diluents, binding agents, lubricants, disintegrants, colorants, stabilizers and surfactants. Diluents, also called "excipients", are generally needed to increase the mass of a solid dosage form to obtain an acceptable size for compressing tablets or forming spheres and granules. Suitable diluents include, but are not limited to, dicalcium phosphate dihydrate, calcium sulfate, lactose, sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, kaolin, sodium chloride, dry starch, hydrolyzed starches, pregelatinized starch, silicon dioxide, titanium oxide, magnesium aluminosilicate and powdered sugar.

Binding agents are used to impart binding properties to a solid dosage formulation and thus ensure that the tablet, sphere or granule remains intact after formation into dosage forms. Suitable binders include, but are not limited to, starch, pregelatinized starch, gelatin, sugars (including sucrose, glucose, dextrose, lactose and sorbitol), polyethylene glycol, waxes, natural and synthetic gums such as gum acacia, tragacanth, sodium alginate, cellulose, including hydroxypropyl methylcellulose, hydroxypropylcellulose, ethylcellulose and vigum; and synthetic polymers such as acrylic acid-methacrylic acid copolymers, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, polyacrylic acid/polymethacrylic acid and polyvinylpyrrolidone.

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

Disintegrants are used to facilitate dosage form disintegration or "disintegration" after administration and typically include, but are not limited to, starch, sodium starch glycolate, sodium carboxymethyl starch, sodium carboxymethyl cellulose, hydroxypropyl cellulose, pregelatinized starch, clays, cellulose, alginine, resins or cross-linked polymers , such as cross-linked PVP (Polyplasdone® XL from GAF Chemical Corp).

Stabilizers are used to inhibit or slow down drug degradation reactions, which include, for example, oxidative 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).

Oral dosage forms such as capsules, tablets, solutions and suspensions can be formulated for controlled release. For example, one or more compounds and optionally one or more additional active agents may be formulated as nanoparticles, microparticles, and combinations thereof and enclosed in a soft or hard gelatin or non-gelatin capsule or dispersed in a dispersant medium to form an oral suspension or syrup. These particles may be formed from 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 finished dosage form.

In another embodiment of the present invention, one or more compounds and optionally one or more additional active agents are dispersed in a matrix material that gels or emulsifies upon contact with an aqueous medium, such as bodily fluids. In the case of gels, the matrix swells, trapping active agents, which are released slowly over time as a result of diffusion and/or degradation of the matrix material. Such matrices can be formulated as tablets or fillers for hard and soft capsules.

In yet another embodiment of the present invention, one or more compounds and optionally one or more additional active agents are included in a marketed/solid oral dosage form, such as a tablet or capsule, wherein one or more coatings are applied to the solid dosage form with controlled release, such as sustained release coatings or sustained release coatings. The coating or coatings may also contain these compounds and/or additional active agents.

Extended-release dosage forms

Sustained release compositions are typically formulated as diffusion or osmotic systems, which are known in the art. A diffusion system typically consists of two types of devices, a reservoir and a matrix, and is well known and described in the art. Matrix devices are typically prepared by compressing a drug with a slowly dissolving polymer carrier into tablet form. The three main types of materials used in the manufacture of matrix devices are insoluble plastics, hydrophilic polymers, and fatty compounds. Plastic matrices include, but are not limited to, methyl acrylate, methyl methacrylate, polyvinyl chloride, and polyethylene. Hydrophilic polymers include, but are not limited to, cellulosic polymers such as methyl and ethyl cellulose, hydroxyalkyl celluloses such as hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose and Carbopol® 934, polyethylene oxides, and mixtures thereof. Fatty compounds include, but are not limited to, various waxes, such as carnauba wax and glyceryl tristearate, and wax-type substances, including hydrogenated castor oil or hydrogenated vegetable oil, or mixtures thereof.

In some preferred embodiments of the present invention, the plastic material is a pharmaceutically acceptable acrylic polymer, including, but not limited to, acrylic acid-methacrylic acid copolymers, methyl methacrylate, methyl methacrylate copolymers, ethoxyethyl methacrylates, cyanoethyl methacrylate, aminoalkyl methacrylate copolymer, poly(acrylic acid), poly( methacrylic acid), methylacrylic acid-alkylamine copolymer, poly(methyl methacrylate), poly(methacrylic acid) (anhydride), polymethacrylate, polyacrylamide, poly(methacrylic acid anhydride) and glycidyl methacrylate copolymers. In certain preferred embodiments of this invention, the acrylic polymer is composed of one or more ammonium methacrylate copolymers. Ammonium methacrylate copolymers are well known in the art and are described in NF XVII as fully polymerized copolymers of acrylic and methacrylic acid esters with a low content of quaternary ammonium groups.

In one preferred embodiment of the present invention, the acrylic polymer is an acrylic synthetic resin varnish, such as that commercially available from Rohm Pharma under the trade name Eudragit®. In further preferred embodiments of the present invention, the acrylic polymer comprises a mixture of two acrylic synthetic resin varnishes commercially available from Rohm Pharma under the trade names Eudragit® RL30D and Eudragit® RS30D, respectively. Eudragit RL30D (Eudragit® RL30D) and Eudragit RS30D (Eudragit® RS30D) are copolymers of acrylic and methacrylic esters with a low content of quaternary ammonium groups, while the molar ratio of ammonium groups to the remaining neutral (meth)acrylic esters is 1:20 in Eudragit RL30D (Eudragit® RL30D) and 1:40 in Eudragit RS30D (Eudragit® RS30D). The average molecular weight is about 150,000. Eudragit® S-100 and Eudragit® L-100 are also preferred. The code designations RL (high permeability) and RS (low permeability) refer to the permeability properties of these agents. Eudragit® RL/RS mixtures are insoluble in water and digestive juices. However, systems consisting of many particles, formed to include these particles, are swelling and permeable in aqueous solutions and digestive juices. The polymers described above, such as Eudragit® RL/RS, can be mixed together in any desired ratio to ultimately produce a sustained release formulation having the desired dissolution profile. Desired multiparticulate sustained release systems can be prepared from, for example, 100% Eudragit®RL, 50% Eudragit® RL and 50% Eudragit®RS, as well as 10% Eudragit RL (Eudragit®RL) and 90% Eudragit RS (Eudragit®RS). One skilled in the art will appreciate that other acrylic polymers can also be used, such as, for example, Eudragit®L.

Alternatively, sustained release formulations can be prepared using osmotic systems or by applying a semi-permeable coating to the dosage form. In the latter case, the desired drug release profile can be achieved by combining low and high permeability coating materials in a suitable proportion.

Devices with different drug release mechanisms described above can be combined into a final dosage form containing one or multiple units. Examples of multiple units include, but are not limited to, multilayer tablets and capsules containing tablets, spheres or granules, etc. An immediate release portion can be added to a sustained release system by applying an immediate release layer over a sustained release core with using a coating or compression process, or into a multi-unit system such as a capsule containing sustained and immediate release spheres.

Extended release tablets containing hydrophilic polymers are prepared using methods well known in the art, such as direct compression, wet granulation or dry granulation processes. Their formulations typically include polymers, diluents, binders and lubricants, and an active pharmaceutical ingredient. Common diluents include inert powders such as starches, powdered cellulose, especially crystalline and microcrystalline cellulose, sugars such as fructose, mannitol and sucrose, cereal flour and similar food powders. Typical diluents include, for example, various types of starch, lactose, mannitol, kaolin, calcium phosphate or sulfate, inorganic salts such as sodium chloride and powdered sugar. Powdered cellulose derivatives are also suitable. Typical tablet binders include substances such as starch, gelatin and sugars such as lactose, fructose and glucose. Natural and synthetic resins can also be used, including gum acacia, alginates, methylcellulose and polyvinylpyrrolidone. Polyethylene glycol, hydrophilic polymers, ethylcellulose and waxes can also be used as binding agents. A lubricating agent is necessary in the tablet formulation to prevent the tablet and punch from getting stuck in the die. The lubricant is selected from slippery solids such as talc, magnesium calcium stearate, stearic acid, and hydrogenated vegetable oils. Extended release tablets containing waxy materials are typically prepared using methods known in the art, such as the direct mixing method , crystallization method and aqueous dispersion method. In the crystallization method, the drug is mixed with a waxy material and either crystallized by spraying or crystallized, sieved and processed.

Sustained release dosage forms

Sustained release formulations can be created 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.

Sustained release dosage forms can be prepared, for example, by coating the drug or composition containing the drug with a selected coating material. Said drug composition may be, for example, a tablet for administration in a capsule, a tablet for use as the inner core in a "coated core" dosage form, or a plurality of drug-containing spheres, particles or granules for inclusion either in a tablet or in capsule. Preferred coating materials include biodegradable, progressively hydrolysable, progressively water-soluble and/or enzymatically degradable polymers and may be conventional "enteric" polymers. Enteric polymers, as will be appreciated by those skilled in the art, become soluble in the higher pH environment of the lower gastrointestinal tract or are slowly degraded as the dosage form passes through the gastrointestinal tract, while enzymatically degradable polymers are degraded by bacterial enzymes. , present in the lower gastrointestinal tract, especially the colon. Suitable coating materials used to achieve sustained release include, but are not limited to, cellulose polymers such as hydroxypropylcellulose, hydroxyethylcellulose, hydroxymethylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose acetate succinate, hydroxypropylmethylcellulose phthalate, methylcellulose, ethylcellulose, cellulose acetate, cellulose acetate phthalate, trimellitate cellulose acetate and sodium carboxymethylcellulose; polymers and copolymers of acrylic acid, preferably formed from acrylic acid, methacrylic acid, methyl acrylate, ethyl acrylate, methyl methacrylate and/or ethyl methacrylate and other methacrylic resins commercially available under the trade name Eudragit® (Rohm Pharma; Westerstadt, Germany) including Eudragit ( Eudragit®) L30D-55 and L100-55 (soluble at pH 5.5 and above), Eudragit® L-100 (soluble at pH 6.0 and above), Eudragit® S (soluble at pH 7.0 and above, resulting in a higher degree of esterification) and Eudragit® NE, RL and RS (water-insoluble polymers having varying degrees of permeability and extensibility); vinyl polymers and copolymers such as polyvinylpyrrolidone, vinyl acetate, vinyl acetate phthalate, vinyl acetate-crotonic acid copolymer, and ethylene-vinyl acetate copolymer; enzymatically degradable polymers such as azopolymers, pectin, chitosan, amylose and guar gum; zein and shellac. Combinations of different coating materials can also be used. It is also possible to apply multilayer coatings using different polymers.

Those skilled in the art can easily determine the preferred coating weight for specific coating materials by evaluating individual release profiles for tablets, spheres, and granules formulated with varying amounts of different coating materials. It is the combination of materials, method and form of application that gives the desired release characteristics, which can only be determined from clinical studies.

The coating composition may include conventional additives such as plasticizers, pigments, dyes, stabilizers, glidants, etc. A plasticizer is usually present to reduce the brittleness of the coating, typically ranging from about 10 wt. % up to 50 wt. % relative to 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. A stabilizing agent is preferably used to stabilize the particles in the dispersion. Typical stabilizing agents are nonionic emulsifiers such as sorbitol esters, polysorbates and polyvinylpyrrolidone. Slip agents are recommended to reduce the sticking effect during film formation and drying and typically range from about 25 wt. % up to 100 wt. % by weight of the polymer in the coating solution. One of the effective slip agents is talc. Other glidants such as magnesium stearate and glycerol monostearates may also be used. Pigments such as titanium dioxide can also be used. Small amounts of an antifoam agent such as silicone (eg simethicone) may also be added to the coating composition.

With. Compositions for administration into the lungs and mucous membranes.

The active agent(s) and compositions thereof can be formulated for pulmonary and mucosal administration. Administration may include delivery of the composition to the lungs, nasal mucosa, oral cavity (sublingual, buccal), vaginal or rectal.

In one embodiment of the present invention, said compounds are formulated for delivery to the lungs, for example, by intranasal administration or oral inhalation. The respiratory tract is an anatomical structure involved in the exchange of gases between the atmosphere and the bloodstream. The lungs are branching anatomical structures that ultimately end in alveoli, where gases are exchanged. The alveolar surface area is the largest in the respiratory system and represents the site where drug absorption occurs. The alveoli are covered with a thin epithelium without cilia or a mucous layer and secrete surface-active phospholipids. The term "airway" covers the upper respiratory tract, including the oropharynx and larynx, followed by the lower respiratory tract, which includes the trachea followed by bifurcation into the bronchi and bronchioles. The upper and lower airways are called the conducting airways. The terminal bronchioles then divide into respiratory bronchioles, which then lead to the terminal respiratory zone, the alveoli or deep lungs. The deep lung, or alveoli, are the primary target for inhaled therapeutic aerosols during systemic drug delivery.

There is administration into the lungs of therapeutic compositions consisting of small molecule drugs, for example, beta-androgen antagonists for the treatment of asthma. Other therapeutic agents that are active in the lungs are administered systemically and act on the target via pulmonary absorption. Nasal delivery is considered a promising route for administering therapeutics for the following reasons: the nose has a large surface area available for drug absorption due to the epithelial surface being covered with numerous microvilli, in addition, the subepithelial layer is highly vascularized, venous blood from the nose enters directly into the systemic circulation and thus prevents the loss of the drug as a result of first-pass metabolism in the liver; such delivery involves the use of lower doses, there is a faster achievement of therapeutic blood levels, a faster onset of pharmacological activity, fewer side effects, high total blood flow per cm ³ , the presence of a porous endothelial basement membrane, and the delivery is easily accessible.

As used herein, the term "aerosol" refers to any preparation of a fine particle composition that may be in solution or suspension, whether or not using a propellant. Aerosols can be produced using standard methods such as ultrasonic or high pressure processing.

Carriers for pulmonary compositions can be divided into carriers for dry powder formulations and carriers for administration in the form of solutions. Aerosols for delivering therapeutic agents to the respiratory tract are known in the art. For administration through the upper respiratory tract, the composition may be formulated in the form of a solution, for example, water or isotonic saline solution, buffered or unbuffered, or in the form of a suspension for intranasal administration in the form of drops or spray. Preferably, such solutions or suspensions are isotonic with respect to nasal secretions and have approximately the same pH value, for example, from about pH 4.0 to about pH 7.4 or from pH 6.0 to pH 7.0. Buffers should be physiologically compatible and include, for example, phosphate buffers. For example, a typical nasal decongestant is described as being buffered to a pH of about 6.2. One skilled in the art can readily determine the appropriate saline content and pH to obtain a harmless aqueous solution for nasal and/or upper respiratory tract administration.

Preferably, the aqueous solution is water, physiologically acceptable aqueous solutions containing salts and/or buffers, such as phosphate buffered saline (PBS), or any other aqueous solution suitable for administration to an animal or human. Such solutions are well known to one skilled in the art and include, but are not limited to, distilled water, deionized water, pure or ultrapure water, saline, phosphate buffered saline (PBS). Other suitable aqueous vehicles include, but are not limited to, Ringer's solution and isotonic sodium chloride solution. Aqueous suspensions may include suspending agents such as cellulose derivatives, sodium alginate, polyvinylpyrrolidone and gum tragacanth, and a wetting agent such as lecithin. Suitable preservatives for aqueous suspensions include ethyl and n-propyl p-hydroxybenzoate.

In another embodiment of the present invention, solvents that are low toxicity organic (ie, non-aqueous) Class 3 residual solvents, such as ethanol, acetone, ethyl acetate, tetrahydrofuran, ethyl ether and propanol, can be used for the formulations. The said solvent is selected based on its ability to easily aerosolize the composition. The reaction involving the specified solvent and compounds should not have harmful consequences. A suitable solvent that dissolves the compounds or forms a suspension of the compounds should be used. The solvent must be sufficiently volatile to form an aerosol solution or suspension. Additional solvents or aerosol agents such as CFCs may be added as desired to increase the volatility of the solution or suspension.

In one embodiment of this invention, the compositions may contain minor amounts of polymers, surfactants or other excipients well known to those skilled in the art. In this context, "negligible amounts" means that there are no excipients that may influence or mediate the absorption of the compounds in the lungs, and that the excipients present are present in an amount that does not adversely affect the absorption of the compounds in the lungs.

Dry lipid powders can be directly dispersed in ethanol due to their hydrophobic nature. For lipids stored in organic solvents such as chloroform, the desired amount of solution is placed in a vial and the chloroform is evaporated under a stream of nitrogen to form a dry thin film on the surface of the glass vial. The film swells easily when reduced with ethanol. In order to completely disperse the lipid molecule in an organic solvent, the suspension is treated with ultrasound. Nonaqueous lipid suspensions can also be prepared in absolute ethanol using a PARI LC Jet+ refillable nebulizer (PARI Respiratory Equipment, Monterey, CA).

Dry powder formulations ("DPFs") with larger particle sizes have improved flow characteristics such as less aggregation, easier aerosolization, and potentially less phagocytosis. Dry powder aerosols for inhalation therapy are typically produced with average diameters, generally in the range of less than 5 microns, although the preferred range is from one to ten microns in aerodynamic diameter. Large "carrier" particles (not containing drug) are delivered along with therapeutic aerosols to help achieve effective aerosolization among other possible benefits.

The polymer particles can be prepared by single and double evaporation of a highly volatile solvent in an emulsion medium, 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 be prepared using methods for making microspheres or microcapsules known in the art. Preferred manufacturing methods are spray drying and freeze drying, which entail using a solution containing a surfactant, spraying to form droplets of the desired size, and removing the solvent.

These particles can be manufactured using appropriate material, surface roughening, and a specific diameter and tapped density for localized delivery to selected areas of the respiratory tract, such as the deep lungs or upper respiratory tract. For example, higher density particles or larger particles may be used for delivery to the upper respiratory tract. Likewise, a mixture of particles of different sizes equipped with the same or different EGS can be administered to target different regions of the lung in a single administration.

Formulations for pulmonary delivery include unilamellar phospholipid vesicles, liposomes, or lipoprotein particles. Compositions and methods for preparing such nucleic acid compositions are well known to one skilled in the art. Liposomes are formed from commercially available phospholipids available from various suppliers, including Avanti Polar Lipids, Inc. (Birmingham, Alabama). In one embodiment of the present invention, the liposome may include a ligand molecule specific for a receptor on the surface of a target cell to direct the liposome to the target cell.

d. Transdermal administration

Formulations for transdermal administration can also be prepared. Usually these are ointments, lotions, sprays or patches that can be prepared using standard technology. Formulations for transdermal administration may include penetration enhancers.

III. Methods for selective activation of Akt3

The described compositions for selective activation of Akt3 can be used to modulate the immune response by reducing the suppressive function of nTregs. In some embodiments of the present invention, compounds of Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, are administered systemically. In other embodiments of the present invention, compounds of Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, are administered topically or regionally. For example, in some embodiments of the present invention, compositions containing compounds of Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, deliver or specifically target tissue or organs in need of modulation. Tregs can be modulated by targeting or delivering compositions to lymph nodes. nTregs can be modulated by targeting or specifically delivering compositions to the thymus or spleen. iTregs can be modulated by targeting or specifically delivering compositions to generic T cells outside the thymus.

In some in vivo approaches, the compositions described herein are administered to a subject in a therapeutically effective amount. As used herein, the term "effective amount" or "therapeutically effective amount" means a dose sufficient to treat, suppress or ameliorate one or more symptoms of the disorder being treated, or to otherwise provide a desired pharmacological and/or physiological effect. The exact dosage will vary depending on many factors, such as subject-specific variables (eg, age, immune system status, etc.), disease, and treatment being administered. Typical symptoms, pharmacological and physiological effects are discussed in more detail below.

In some embodiments of the present invention, the effect of the composition on a subject is compared with a control. For example, the effect of the composition on a particular symptom, pharmacological or physiological parameter can be compared with the subject not receiving treatment or the condition of the subject before treatment. In some embodiments of the present invention, a symptom, pharmacological or physiological indicator is assessed in the subject before treatment and thereafter one or more times after the start of treatment. In some embodiments of the present invention, the control is a reference level or average value determined based on the measurement of a symptom, pharmacological or physiological indicator in one or more subjects who do not have the disease or condition being treated (eg, healthy subjects). In some embodiments of the present invention, the effect of the treatment is compared with conventional treatment known in the art. For example, if the disease to be treated is cancer, conventional treatment may be the use of a chemotherapeutic agent.

In some embodiments of this invention, the immunomodulatory compositions described herein are administered in combination with one or more additional active agents. Combination therapy may involve administering the active agents together in the same mixture or in separate mixtures. Therefore, in some embodiments of the present invention, said pharmaceutical composition includes two, three or more active agents. Pharmaceutical compositions can be formulated in a pharmaceutical unit dosage form, called a unit dosage form. Such formulations typically include an effective amount of one or more of the disclosed immunomodulatory compounds. Different active agents may have the same or different mechanisms of action. In some embodiments of the present invention, the combination results in an additive effect in treating the disease or disorder. In some embodiments of the present invention, the combinations result in a more than additive effect in treating the disease or disorder.

Preferably, the described compounds and methods of use specifically activate Akt3 activity without increasing or decreasing the activity of Akt1, Akt2, or a combination thereof.

A. Strengthening immunosuppressive reactions and weakening immunostimulating reactions

1. Treatment methods

The described compounds or their enantiomer, polymorph or pharmaceutically acceptable salt are useful as therapeutic agents. Immune cells, preferably T cells, can be contacted in vivo or ex vivo with compounds according to Formula I, Formula II, Formula III or Formula IV or an enantiomer, polymorph or pharmaceutically acceptable salt thereof to reduce or inhibit immune responses, including inflammation, but not limited to them. The T cells contacted by the compounds of Formula I, Formula II, Formula III or Formula IV, or an enantiomer, polymorph or pharmaceutically acceptable salt thereof, can be any immune cell that expresses Akt3 or has Akt3 activity and has the ability to become Foxp3+. Exemplary immune cells that can be treated with compounds of Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof include, but are not limited to, regulatory cells such as ThI cells, TcI Th25 cells , Tc2, Th3, ThI 7, Th22, Treg, nTreg, iTreg and TrI, as well as cells that secrete or induce the secretion by other cells of inflammatory molecules, including, but not limited to, IL-Iβ, TNF-α TGF-beta, IFN-γ, IL-17, IL-6, IL-23, IL-22, IL-21 and MMP. The described compounds, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, can also be used to increase or stimulate Treg activity or production, increase the production of cytokines such as IL-10 from Treg, increase Treg differentiation, increase Treg number, or increase Treg survival.

The described compounds or their enantiomer, polymorph or pharmaceutically acceptable salt can be used to increase the expression of FoxP3 on immune cells.

In one embodiment, the present invention provides a method of enhancing an immune suppressive response in a subject in need thereof by ex vivo contacting immune cells with the disclosed compounds or an enantiomer, polymorph or pharmaceutically acceptable salt thereof in an amount effective to increase FoxP3 expression on the immune cells , and introducing the contacted immune cells to said subject. In one embodiment of the present invention, said immune cells are autologous immune cells. Said immune cells may include T cells, including, but not limited to, Tregs and iTregs.

In some embodiments of this invention, the described compounds, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, are administered in combination with a second therapeutic agent. In immune modulation, combination therapy may be beneficial. In some embodiments of the present invention, the described compounds, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, can be used to attenuate or reverse the activity of a proinflammatory drug and/or limit the undesirable effects of such drugs.

B. Methods for treating inflammatory reactions

In one embodiment, the present invention provides methods for treating or alleviating one or more symptoms of inflammation. In a more preferred embodiment of the present invention, compositions according to Formula I, Formula II, Formula III or Formula IV and the methods described are useful for the treatment of chronic and persistent inflammation. Inflammation in general can be treated using compounds of Formula I, Formula II, Formula III or Formula IV or an enantiomer, polymorph or pharmaceutically acceptable salt thereof.

The immune response, including inflammation, can be inhibited or attenuated in a subject, preferably a human, by administering an effective amount of the disclosed compounds or an enantiomer, polymorph or pharmaceutically acceptable salt thereof to increase or stimulate the biological activity of Akt3 in an immune cell, reduce the number of pro-inflammatory molecules at the site of inflammation , induce or increase FoxP3 expression, induce or increase iTreg proliferation, or to achieve combinations of these effects. Typical proinflammatory molecules include, but are not limited to, IL-1 β, TNF-α, TGF-β, IFN-γ, IL-17, IL-6, IL-23, IL-22, IL-21 and MMP.

Compounds according to Formula I, Formula II, Formula III or Formula IV, or an enantiomer, polymorph or pharmaceutically acceptable salt thereof, may cause an enhanced suppressive effect of Tregs on the immune response. Tregs can suppress the differentiation, proliferation, activity and/or production and/or secretion of cytokines by Th1, Th17, Th7, Th22 cells and/or other cells that secrete or induce the secretion by other cells of inflammatory molecules, including, but not limited to, IL- 1 β, TNF-α TGF-beta, IFN-γ, IL-17, IL-6, IL-23, IL-22, IL-21 and MMP. For example, compounds according to Formula I, Formula II, Formula III or Formula IV, or an enantiomer, polymorph or pharmaceutically acceptable salt thereof, can cause an enhanced suppressive effect of Treg on Th1 and/or Th 17 cells to reduce the level of IFN-γ and IL-γ produced. 17, respectively. The disclosed compounds, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, may also act directly on Tregs to stimulate or enhance IL-10 production, suppress the Th1 and Th17 pathway, or increase the number of Tregs.

1. Diseases to be treated

Compositions containing the described compounds or an enantiomer, polymorph or pharmaceutically acceptable salt thereof that selectively increase Akt3 activity or expression can be used to reduce the immunostimulatory response in a subject. In some embodiments of the present invention, the subjects have an inflammatory disease, including, but not limited to, an autoimmune disease.

Representative inflammatory or autoimmune diseases and disorders that can be treated with the disclosed compounds or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, or with compositions containing the disclosed compounds or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof include, but are not limited to, rheumatoid arthritis, systemic lupus erythematosus, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome (ALPS), autoimmune thrombocytopenic purpura (ATP), Behçet's disease, bullous pemphigoid , cardiomyopathy, dermatitis associated with celiac spruce, chronic fatigue and immunodeficiency syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, cicatricial pemphigoid, cold agglutinin disease, Cross syndrome, Crohn's disease, Dego's disease, dermatomyositis, juvenile dermatomyositis, discoid lupus , essential mixed cryoglobulinemia, fibromyalgia - fibromyositis, Graves' disease, Guillain-Barre disease, Hashimoto's thyroiditis, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA nephropathy, insulin-dependent diabetes (type I), juvenile arthritis, Meniere's disease, mixed disease connective tissue, multiple sclerosis, myasthenia gravis, obesity, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, Raynaud's phenomenon, Reiter's syndrome, rheumatic fever ku, sarcoidosis , scleroderma, Sjogren's syndrome, stiff muscle syndrome, Takayasu arteritis, temporal arteritis/giant cell arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo and Wegener's granulomatosis.

2. Combination therapeutic agents

The described compounds or their enantiomer, polymorph or pharmaceutically acceptable salt and compositions thereof can be used alone or in combination with additional therapeutic agents. The described compounds can be administered together or alternately with additional therapeutic agents. Additional therapeutic agents include, but are not limited to, immunosuppressive agents (e.g., antibodies against other lymphocyte surface markers (e.g., CD40, alpha-4 integrin) or against cytokines), other fusion proteins (e.g., CTLA-4-Ig, abatacept ( Orencia®), TNF-α blockers such as TNFR-Ig, etanercept (Enbrel®), infliximab (Remicade®), certolizumab (Cimzia®), and adalimumab (Humira ®), cyclophosphamide (CTX) (i.e. Endoxan®, Cytoxan®, Neosar®, Procytox®, Revimmune™), methotrexate (MTX) (i.e. Rheumatrex®, Trexall®), belimumab (ie Benlysta®) or other immunosuppressive drugs (eg cyclosporine A, FK506-like compounds, rapamycin compounds or steroids), antiproliferative agents, cytotoxic agents or others compounds that may contribute to immunosuppression.

Additional immunosuppressive agents include, but are not limited to, prednisone, budesonide, prednisolone, cyclosporine, tacrolimus, sirolimus, everolimus, azathioprine, leflunomide, mycophenolate, anakinra, golimumab, ixekizumab, natalizumab, rituximab, secukinumab, tocilizumab, ustekinumab, vedolizum ab, basiliximab, daclizumab, muromonab, or combinations thereof.

In one embodiment, the present invention provides an additional therapeutic agent that functions to inhibit or reduce T cell activation through a distinct pathway. In one such embodiment of the present invention, the additional therapeutic agent is a CTLA-4 fusion protein, such as CTLA-4-Ig (abatacept). CTLA-4-Ig fusion proteins compete with the costimulatory receptor, CD28, on T cells for binding to CD80/CD86 (B7-1/B7-2) on antigen presenting cells and thus function to inhibit T cell activation. In another embodiment of the present invention, the additional therapeutic agent is a CTLA-4-Ig fusion protein known as belatacept. Belatacept contains two amino acid substitutions (L104E and A29Y) that markedly increase its avidity for CD86 in vivo. In another embodiment of the present invention, the additional therapeutic agent is Maxy-4.

In another embodiment of the present invention, the second therapeutic agent is cyclophosphamide (CTX). Cyclophosphamide (generic name for

Endoxan®, Cytoxan®, Neosar®, Procytox®, Revimmune™), also known as cytophosphan, is an alkylating agent of mustargen from the oxazophorine group. The drug is used to treat various types of cancer and some autoimmune diseases. In another embodiment of the present invention, compounds of Formula I, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, and CTX are administered together in an effective amount to inhibit, alleviate symptoms of, or treat a chronic autoimmune disease or disorder, such as systemic lupus erythematosus (SLE).

In another embodiment of the present invention, the second therapeutic agent is preferably used to treat chronic inflammation, wherein the treatment regimen targets both acute and chronic inflammation.

In another embodiment of the present invention, compositions according to Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, are used in combination, in alternation, or in sequence with compounds that enhance Treg activity or production. Typical Treg enhancing agents include, but are not limited to, the glucocorticoid fluticasone, salmeterol, antibodies to IL-12, IFN-γ, and IL-4; vitamin D3 and dexamethasone, as well as their combinations.

Antibodies to other pro-inflammatory molecules can also be used in combination or alternation with the described compounds according to Formula I, Formula II, Formula III or Formula IV or an enantiomer, polymorph or pharmaceutically acceptable salt, fusion proteins or fragments thereof. Preferred antibodies bind to IL-6, IL-23, IL-22 or IL-21.

In another embodiment, the present invention provides a method of treating transplant rejection by administering to a subject in need thereof an effective amount of the described compounds according to Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, to increase expression of FoxP3 on immune cells.

In another embodiment, the present invention provides a method of treating graft-versus-host disease by administering to a subject in need thereof an effective amount of a disclosed compound of Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof. to increase FoxP3 expression on immune cells.

In yet another embodiment, the present invention provides a method of inhibiting or reducing transplant rejection in a host in need thereof by administering to the subject in need thereof an effective amount of the disclosed compounds or an enantiomer, polymorph or pharmaceutically acceptable salt thereof to increase FoxP3 expression on immune cells.

In another embodiment, the present invention provides a method of treating a chronic infection by administering to a subject in need thereof an effective amount of the described compounds according to Formula I, Formula II, Formula III, or Formula IV, or an enantiomer, polymorph, or pharmaceutically acceptable salt thereof, to increase expression of FoxP3 on immune cells.

In one embodiment, the present invention provides a method of treating obesity by administering to a subject in need thereof an effective amount of the described compounds according to Formula I, Formula II, Formula III or Formula IV, or an enantiomer, polymorph or pharmaceutically acceptable salt thereof, for the purpose of increasing Akt3 activity . Without wishing to be bound by any one theory, it is believed that Akt3 regulates adipogenesis and that dysregulation of Akt3 signaling may lead to increased adipogenesis, obesity, and insulin resistance.

IV. Sets

This document also describes medical kits. Medical kits may include, for example, providing a dose of the disclosed compounds or an enantiomer, polymorph or pharmaceutically acceptable salt thereof or compositions thereof. The described compounds or their enantiomer, polymorph or pharmaceutically acceptable salt or compositions thereof may be supplied separately (eg, lyophilized) or in a pharmaceutical composition. The described compounds or their enantiomer, polymorph or pharmaceutically acceptable salt or compositions thereof may be in a unit dose or in a volume that must be diluted before administration. In some embodiments of the present invention, the kit includes providing a pharmaceutically acceptable carrier. The kit may also include devices for administering the active agent(s) or composition(s), such as syringes and needles. These kits may include printed instructions for administration of the described compounds when used as described above.

Examples

Example 1: mJJ64A significantly increases FoxP3 expression on iTregs.

results

The findings demonstrate that mJJ64A significantly increased FoxP3 expression on iTregs and nonsignificantly increased iTreg proliferation ( Figures 2A-2H ).

Example 2: mJJ64A increases FoxP3 expression when added during iTreg induction.

results

The findings demonstrate that mJJ64A increases FoxP3 expression when added during iTreg induction (Figures 3A and 3B).

Example 3: Akt3 specifically regulates both types of Tregs: nTreg and iTreg.

results

The findings demonstrate that Akt3 is a key regulator of nTregs (Figures 4A-4F). The suppressive activity of Tregs in Akt3 KO mice was due to decreased levels of the inhibitory cytokines IL-10 and TGFβ ( Figures 4E and 4F ). The data also demonstrate that in the absence of Akt3, but not other isoforms, Treg suppressive activity was impaired in vivo (Figures 5A-5B). Tregs from Akt3 KO mice showed impaired suppressive activity in the RAG colitis model (Figure 5A). Additionally, adoptive transfer of Tregs from Akt3 KO mice to Treg-depleted tumor mice demonstrates impaired antitumor immune suppression ( Figure 5B ).

The findings also demonstrated that Akt3 was a key regulator of iTregs ( Figures 6A–6E ). Akt3 RNA, protein, and Akt3 phosphorylation were upregulated in iTregs ( Figures 6A–6C ). In Akt3 KO mice, the conversion of Tconv cells to iTregs was significantly inhibited ( Figure 6D ). In addition, knockdown of Akt3 from WT Tconv cells abolished FoxP3 induction in response to TGFβ (Figure 6 E).

Fig. 7A-I demonstrate that Akt3 knockin was sufficient to induce Tregs, as demonstrated by FoxP3 activation.

Example 4: mJJ64A increases Akt3 phosphorylation in human ovarian carcinoma cells.

results

Findings demonstrate that mJJ64A significantly increases phosphorylation of Akt3, but not Akt1 or Akt2, in human ovarian carcinoma cells (Figure 8)

Example 5: mJJ64A enhances FoxP3 and Akt3 in Tconv cells during iTreg induction.

results

The findings demonstrate that mJJ64A exposure increased FoxP3 and Akt3 expression in Tconv cells during iTreg induction (Figures 9A-9B).

Example 6: mJJ64A increases iTreg and nTreg proliferation

results

mJJ64A exposure increased the proliferation of iTreg (FIGS. 10A - 10C) and nTreg (FIGS. 10D - 10F), but non-Treg CD4 (FIGS. 10G - 10I) and CD8 (FIGS. 10J - 10L) T cells.

Example 7: mJJ64A enhances the suppressive function of mouse iTregs and nTregs.

results

In FIG. 11A - 11D demonstrate that mJJ64A enhanced the suppressive function of mouse iTreg cells in vitro . mJJ64A also increased the suppressive function of mouse nTreg cells in vitro and increased nTreg proliferation without affecting their viability (Figures 12A to 12H).

Example 8: mJJ64A enhances IL-10 production by nTregs.

results

The findings demonstrate that IL-10 production by nTreg cells was increased by mJJ64A exposure (FIGS. 13A - 13B).

Example 9: mJJ64A increases TC-1 tumor growth and significantly increases Tregs in tumors and spleens of treated mice.

results

The findings demonstrate that mice bearing TC-1 tumors treated with mJJ64A experienced significantly greater tumor growth compared to untreated controls (Figures 14A and 14B). mJJ64A also increased the number of Tregs in tumors and spleens of treated mice compared to untreated controls (Figures 15D and 15E). Exposure to mJJ64A had no effect on tumor infiltration of CD8 ⁺ and FoxP3 ^NEG CD4 T cells (Figures 15A - 15C).

Example 10: mJJ64A protects against experimental colitis.

results

The findings demonstrate that exposure to mJJ64A protected against experimental colitis (Figures 16A - 16I and Figures 17A - 17J). In addition, administration of iTreg mice treated with mJJ64A ex vivo also provided protection against experimental colitis (Figures 16A to 16I and Figures 17A to 17J).

Example 11: mJJ64A increases the percentage of Tregs in Rag-/- mice.

results

The findings demonstrate that administration of mJJ64A to Rag −/− mice increased the percentage of Tregs in the spleen and mesenteric lymph nodes compared to untreated Rag −/− mice (Fig. 18A - F).

Example 12 Efficacy of mJJ64A in the EAE mouse model.

results

mJJ64A slowed disease progression and increased survival in an experimental mouse model of autoimmune encephalomyelitis (EAE) (Figures 19A - 19F). In addition, mJJ64A-induced iTregs also slowed disease progression and increased survival in the EAE model compared to untreated controls (Figure 19).

Example 13: mJJ64A enhances iTreg induction without affecting cell viability.

results

The findings demonstrate that mJJ64A induced human iTregs (Figure 21B) but did not affect cell viability (Figure 21A).

--->

LIST OF SEQUENCES

<110> Augusta University Research Institute, Inc

Khleif, Samir N

Mkrtichyan, Mikayel

<120> SPECIFIC ACTIVATOR AKT3 AND ITS APPLICATION

<130> 064466.070

<150> 62/555,141

<151> 2017-09-07

<150> 62/657.345

<151> 2018-04-13

<150> 62/659,870

<151> 2018-04-19

<160> 3

<170> PatentIn version 3.5

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<---

Claims (148)

1. Compound of formula I or a pharmaceutically acceptable salt thereof, Where: rings A, B and C are independently phenyl, pyridine and quinoline; R ₁ represents -(C ₃ -C ₁₂ )-heterocycloalkyl, -(C ₆ -C ₂₀ )-aryl, or -(C ₃ -C ₂₀ )-heteroaryl groups, optionally substituted by one or more substituents selected from - (C ₁ -C ₁₂ )-alkyl or -NH ₂ ; X, Y and Z are independently -O, -NH, or -N-(C ₁ -C ₃₀ )-alkyl; R ₂ represents =O; And R ₃ on ring A is -(C ₁ -C ₃₀ )-alkyl, -(C ₃ -C ₁₂ )-heterocycloalkyl, -(C ₆ -C ₂₀ )-aryl, -O-(C ₁ -C ₁₂ ) -alkyl, -N-[(C ₁ -C ₁₂ )-alkyl] ₂ , -COOH, -CN, -NH ₂ or halogen; R ₃ on ring C is hydrogen, -(C ₁ -C ₃₀ )-alkyl or halogen; provided that (i) when R ₁ is unsubstituted pyridine or N-substituted –(C1)-alkyl pyridine, X, Y and Z are NH, R ₂ is =O, A is quinoline or pyridine, B is phenyl or pyridine, C is phenyl, R ₃ on ring C is hydrogen, then R ₃ on ring A is not -N-[(C ₁ )-alkyl] ₂ , -COOH, -CN and –F; (ii) when the compound has the structure of formula III, R ₁ is unsubstituted pyridine, N-substituted -(C ₁ -C ₃ )-alkyl pyridine or pyrimidine substituted –(C ₁ )-alkyl and NH ₂ , X, Y, Z are -NH, R ₂ is =O, then R ₄ is not a halogen, -(C ₁ )-alkyl, -O-(C ₁ )-alkyl, -NH ₂ and -N-[(C ₁ )-alkyl] ₂ ; (iii) when R₁ is unsubstituted pyridine, unsubstituted quinoline, or quinoline substituted -NH₂, -NO₂, or -N-[(C₁)-alkyl]₂, X, Y, Z represent -NH, R₂ is =O, A is pyridine, B is phenyl, C is phenyl, R₃ on the ring C represents hydrogen, then R₃ on the ring A is not –(C₁-C₂)-alkyl and -NH₂; And (iv) when R ₁ is acridine, X, Y, Z are -NH, R ₂ is =O, A is pyridine, B is phenyl, C is phenyl, R ₃ on ring C is hydrogen , then R ₃ on ring A is not –(C ₁ )-alkyl. 2. Compound of formula II Formula II or a pharmaceutically acceptable salt thereof, where: R ₁ represents -(C ₆ -C ₂₀ )-aryl or -(C ₃ -C ₂₀ )-heteroaryl groups, optionally substituted by one or more substituents selected from -(C ₁ -C ₁₂ )-alkyl; X, Y and Z are independently -O or -NH; R ₂ represents =O; And R ₃ represents -(C ₁ -C ₃₀ )-alkyl, or -NH ₂ , provided that (i) when R ₁ is quinoline substituted with -NH ₂ , -NO ₂ , or -N-[(C ₁ )-alkyl] ₂ , X, Y, Z are -NH, then R ₃ on ring A is not is –(C ₁ -C ₂ )-alkyl and -NH ₂ ; And (ii) when R ₁ is acridine, X, Y, Z are -NH, then R ₃ on ring A is not –(C ₁ )-alkyl. 3. Compound of formula III Formula III or a pharmaceutically acceptable salt thereof, where: R ₁ represents -(C ₃ -C ₁₂ )-heterocycloalkyl, -(C ₆ -C ₂₀ )-aryl or -(C ₃ -C ₂₀ )-heteroaryl groups, optionally substituted by one or more substituents selected from -( C ₁ -C ₁₂ )-alkyl and -NH ₂ ; X, Y and Z are independently -O, -NH, or -N-(C ₁ -C ₃₀ )-alkyl; R ₂ represents =O; And R ₄ represents -(C ₁ -C ₁₂ )-alkyl, -(C ₃ -C ₁₂ )-heterocycloalkyl, -O-(C ₁ -C ₁₂ )-alkyl, -N-[(C ₁ -C ₁₂ ) -alkyl] ₂ , -(C ₆ -C ₂₀ )-aryl, -COOH, -CN, -NH ₂ or halogen, provided that: when R ₁ is unsubstituted pyridine, pyridine N-substituted with -(C ₁ -C ₃ )-alkyl, or pyrimidine substituted with –(C ₁ )-alkyl and -NH ₂ , X, Y, Z are -NH, R ₂ is =O, then _R4 is non-halogen, -CN, -COOH, –( _C1 )-alkyl, -O-( _C1 )-alkyl, _-NH2 , and -N-[( _C1 ) -alkyl] ₂ . 4. A method of enhancing an immunosuppressive response in a subject in need thereof, comprising administering to said subject a compound of formula I Formula I or a pharmaceutically acceptable salt thereof, Where rings A, B and C are independently phenyl, pyridine or quinoline; R ₁ represents -(C ₃ -C ₁₂ )-heterocycloalkyl, -(C ₆ -C ₂₀ )-aryl or -(C ₃ -C ₂₀ )-heteroaryl groups, optionally substituted by one or more substituents selected from -( C ₁ -C ₁₂ )-alkyl and -NH ₂ ; X, Y and Z are independently -O, -NH or -N-(C ₁ -C ₃₀ )-alkyl; R ₂ represents =O; R ₃ on ring A is -(C ₃ -C ₁₂ )-heterocycloalkyl, -N-[(C ₁ -C ₁₂ )-alkyl] ₂ or -CN; And R ₃ on ring C is hydrogen, -(C ₁ -C ₃₀ )-alkyl or halogen; which selectively activates Akt3 in an amount effective to enhance the immunosuppressive response in the specified subject. 5. The method according to claim 4, where the compound is a compound of formula IV: . 6. The method of claim 4 or 5, wherein said subject has an inflammatory disorder or disease. 7. A method of treating an inflammatory disorder, comprising administering to a subject in need thereof a composition containing compounds of formula I Formula I or a pharmaceutically acceptable salt, Where rings A, B and C are independently phenyl, pyridine or quinoline; R ₁ represents -(C ₃ -C ₁₂ )-heterocycloalkyl, -(C ₆ -C ₂₀ )-aryl or -(C ₃ -C ₂₀ )-heteroaryl groups, optionally substituted by one or more substituents selected from -( C ₁ -C ₁₂ )-alkyl and -NH ₂ ; X, Y and Z are independently -O, -NH or -N-(C ₁ -C ₃₀ )-alkyl; R ₂ represents =O; R ₃ on ring A is -(C ₃ -C ₁₂ )-heterocycloalkyl, -N-[(C ₁ -C ₁₂ )-alkyl] ₂ or -CN; And R ₃ on ring C is hydrogen, -(C ₁ -C ₃₀ )-alkyl or halogen, which selectively activate Akt3 in an amount effective to induce, enhance or stimulate an immunosuppressive response in the subject. 8. The method of claim 6 or 7, wherein said inflammatory disorder or disease is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome (ALPS), autoimmune thrombocytopenic purpura (ATP), Behçet's disease, bullous pemphigoid, cardiomyopathy, dermatitis associated with celiac spruce, chronic fatigue and immunodeficiency syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy , cicatricial pemphigoid, cold agglutinin disease, Cross syndrome, Crohn's disease, Dego's disease, dermatomyositis, juvenile dermatomyositis, discoid lupus, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, Graves' disease, Guillain-Barré disease, Hashimoto's thyroiditis, idiopathic pulmonary fibrosis, idiopathic tic thrombocytopenic purpura (ITP), IgA nephropathy, insulin-dependent diabetes (type I), juvenile arthritis, Meniere's disease, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, obesity, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia rheumatica , polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, Raynaud's phenomenon, Reiter's syndrome, rheumatic fever, sarcoidosis, scleroderma, Sjogren's syndrome, muscle stiffness syndrome, Takayasu arteritis, temporal arteritis/giant cell arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo and Wegener's granulomatosis. 9. A method of treating an autoimmune disease, comprising administering to a subject in need thereof a composition containing any of the compounds according to claims. 1-3 or a compound of formula IV specified in claim 5, or a pharmaceutically acceptable salt thereof, which selectively activate Akt3 in an amount effective to induce, stimulate or enhance an immunosuppressive response in the specified subject. 10. The method of claim 9, wherein said autoimmune disease is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus, alopecia areata, autoimmune Addison's disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome (ALPS) ), autoimmune thrombocytopenic purpura (ATP), Crohn's disease, multiple sclerosis and myasthenia gravis. 11. Method according to any one of paragraphs. 4-10, wherein the immunosuppressive response that is enhanced is selected from the group consisting of the immunosuppressive function of a native Treg (nTreg) and the induction of conversion of a typical T cell to an induced Treg (iTreg). 12. The method of claim 11, wherein the immunosuppressive function of the nTreg is the secretion of one or more anti-inflammatory cytokines. 13. The method of claim 12, wherein the anti-inflammatory cytokine is IL-10, TGFβ, or a combination thereof. 14. Method according to any one of paragraphs. 4-13, in which the compound is administered in an amount that increases the expression of FoxP3 on immune cells. 15. The method of claim 14, wherein the immune cells contain iTregs. 16. The method of claim 15, wherein the compound is administered in an amount effective to increase iTreg proliferation. 17. Pharmaceutical composition for selective activation of Akt3 in immune cells, containing an effective amount of a compound of formula I , Where rings A, B and C are independently phenyl, pyridine or quinoline; R ₁ represents -(C ₃ -C ₁₂ )-heterocycloalkyl, -(C ₆ -C ₂₀ )-aryl or -(C ₃ -C ₂₀ )-heteroaryl groups, optionally substituted by one or more substituents selected from -( C ₁ -C ₁₂ )-alkyl and -NH ₂ ; X, Y and Z are independently -O, -NH or -N-(C ₁ -C ₃₀ )-alkyl; R ₂ represents =O; R ₃ on ring A is -(C ₃ -C ₁₂ )-heterocycloalkyl, -N-[(C ₁ -C ₁₂ )-alkyl] ₂ or -CN; And R ₃ on ring C is hydrogen, -(C ₁ -C ₃₀ )-alkyl or halogen; or a pharmaceutically acceptable salt thereof, as well as an excipient. 18. The pharmaceutical composition of claim 17, wherein the compound or a pharmaceutically acceptable salt thereof is in an amount effective to enhance a suppressive immune response when administered to a subject in need thereof. 19. A method of enhancing an immunosuppressive response in a subject in need thereof, comprising ex vivo contacting immune cells with a compound of formula I Formula I or a pharmaceutically acceptable salt thereof, Where rings A, B and C are independently phenyl, pyridine or quinoline; R ₁ represents -(C ₃ -C ₁₂ )-heterocycloalkyl, -(C ₆ -C ₂₀ )-aryl or -(C ₃ -C ₂₀ )-heteroaryl groups, optionally substituted by one or more substituents selected from -( C ₁ -C ₁₂ )-alkyl and -NH ₂ ; X, Y and Z are independently -O, -NH or -N-(C ₁ -C ₃₀ )-alkyl; R ₂ represents =O; R ₃ on ring A is -(C ₃ -C ₁₂ )-heterocycloalkyl, -N-[(C ₁ -C ₁₂ )-alkyl] ₂ or -CN; And R ₃ on ring C is hydrogen, -(C ₁ -C ₃₀ )-alkyl or halogen, in an amount effective to increase FoxP3 expression on immune cells, and administering said contacted immune cells to said subject. 20. The method of claim 19, wherein said immune cells include autologous immune cells. 21. The method of claim 19 or 20, wherein said immune cells include T cells. 22. The method of claim 21, wherein said T cells include Tregs. 23. The method of claim 22, wherein said Tregs include iTregs. 24. A method of inhibiting or reducing transplant rejection in a subject in need thereof by administering an effective amount of a compound of formula I Formula I or a pharmaceutically acceptable salt thereof, Where rings A, B and C are independently phenyl, pyridine or quinoline; R ₁ represents -(C ₃ -C ₁₂ )-heterocycloalkyl, -(C ₆ -C ₂₀ )-aryl, or -(C ₃ -C ₂₀ )-heteroaryl groups, optionally substituted by one or more substituents selected from - (C ₁ -C ₁₂ )-alkyl and -NH ₂ ; X, Y and Z are independently -O, -NH or -N-(C ₁ -C ₃₀ )-alkyl; R ₂ represents =O; R ₃ on ring A is -(C ₃ -C ₁₂ )-heterocycloalkyl, -N-[(C ₁ -C ₁₂ )-alkyl] ₂ or -CN; And R ₃ on ring C is hydrogen, -(C ₁ -C ₃₀ )-alkyl or halogen, to increase the expression of FoxP3 on immune cells of the specified subject. 25. A method of treating graft-versus-host disease in a subject in need thereof by administering to said subject an effective amount of a compound of formula I Formula I or a pharmaceutically acceptable salt thereof, wherein rings A, B and C are independently phenyl, pyridine or quinoline; R ₁ represents -(C ₃ -C ₁₂ )-heterocycloalkyl, -(C ₆ -C ₂₀ )-aryl, or -(C ₃ -C ₂₀ )-heteroaryl groups, optionally substituted by one or more substituents selected from - (C ₁ -C ₁₂ )-alkyl and -NH ₂ ; X, Y and Z are independently -O, -NH or -N-(C ₁ -C ₃₀ )-alkyl; R ₂ represents =O; R ₃ on ring A is -(C ₃ -C ₁₂ )-heterocycloalkyl, -N-[(C ₁ -C ₁₂ )-alkyl] ₂ or -CN; And R ₃ on ring C is hydrogen, -(C ₁ -C ₃₀ )-alkyl or halogen, to increase the expression of FoxP3 on immune cells of the specified subject. 26. A method of treating a chronic infection in a subject in need thereof by administering to said subject an effective amount of a compound of formula I Formula I or a pharmaceutically acceptable salt thereof, wherein rings A, B and C are independently phenyl, pyridine or quinoline; R ₁ represents -(C ₃ -C ₁₂ )-heterocycloalkyl, -(C ₆ -C ₂₀ )-aryl, or -(C ₃ -C ₂₀ )-heteroaryl groups, optionally substituted by one or more substituents selected from - (C ₁ -C ₁₂ )-alkyl and -NH ₂ ; X, Y and Z are independently -O, -NH or -N-(C ₁ -C ₃₀ )-alkyl; R ₂ represents =O; R ₃ on ring A is -(C ₃ -C ₁₂ )-heterocycloalkyl, -N-[(C ₁ -C ₁₂ )-alkyl] ₂ or -CN; And R ₃ on ring C is hydrogen, -(C ₁ -C ₃₀ )-alkyl or halogen, to increase the expression of FoxP3 on immune cells of the specified subject. 27. A method of treating obesity in a subject in need thereof by administering to said subject an effective amount of a compound of formula I Formula I or a pharmaceutically acceptable salt thereof, wherein rings A, B and C are independently phenyl, pyridine or quinoline; R ₁ represents -(C ₃ -C ₁₂ )-heterocycloalkyl, -(C ₆ -C ₂₀ )-aryl, or -(C ₃ -C ₂₀ )-heteroaryl groups, optionally substituted by one or more substituents selected from - (C ₁ -C ₁₂ )-alkyl and -NH ₂ ; X, Y and Z are independently -O, -NH or -N-(C ₁ -C ₃₀ )-alkyl; R ₂ represents =O; R ₃ on ring A is -(C ₃ -C ₁₂ )-heterocycloalkyl, -N-[(C ₁ -C ₁₂ )-alkyl] ₂ or -CN; And R ₃ on ring C is hydrogen, -(C ₁ -C ₃₀ )-alkyl or halogen. 28. The connection according to any one of paragraphs. 1-3 or pharmaceutical composition according to claims. 17 and 18, or the method according to any one of paragraphs. 4-16 and 19-27, where R ₁ in formula I and/or III represents phenyl, pyridine, pyrimidine or pyridazine, optionally substituted with one or more substituents selected from -(C ₁ -C ₁₂ )-alkyl and NH ₂ , and where R ₁ in formula II represents phenyl, pyridine, pyrimidine or pyridazine, optionally substituted by one or more –(C ₁ -C ₁₂ )-alkyl. 29. The connection according to any one of paragraphs. 1-3, 28, or a pharmaceutical composition according to claims. 17, 18 or the method according to any one of paragraphs. 4-16, 19-27, where R ₃ on ring A in formula I and/or R ₄ in formula III independently represents CN, -N-[(C ₁ -C ₁₂ )-alkyl] ₂ or -(C ₁ -C ₁₂ )-heterocycloalkyl. 30. The connection according to any one of paragraphs. 1-3, 28, or a pharmaceutical composition according to claims. 17, 18, or the method according to any one of paragraphs. 9-16, 19-27, where R ₃ on ring A in formula I, R ₃ in formula II, and/or R ₄ in formula III are independently halogen, -(C ₁ -C ₃₀ )-alkyl or NH ₂ .