NZ766879B2

NZ766879B2 - Method of treatment and compositions comprising a dual pi3k delta-gamma kinase inhibitor and a corticosteroid

Info

Publication number: NZ766879B2
Application number: NZ766879A
Authority: NZ
Inventors: Venkata Satya Vakkalanka Swaroop Kumar; Srikant Viswanadha
Original assignee: Rhizen Pharmaceuticals Sa
Priority date: 2014-09-03
Filing date: 2015-09-03
Publication date: 2021-11-30

Abstract

This present disclosure relates to a method of treating autoimmune, respiratory and/or inflammatory diseases or conditions, e.g., asthma, COPD, rheumatoid arthritis and idiopathic Pulmonary Fibrosis (IPF). The method comprises administering a dual PI3K delta and gamma inhibitor and a corticosteroid, wherein the corticosteroid is selected from dexamethasone, fluticasone, fluticasone propionate and mometasone furoate. The present invention also relates to pharmaceutical compositions containing a dual PI3K delta and gamma inhibitor and a corticosteroid. wherein the corticosteroid is selected from dexamethasone, fluticasone, fluticasone propionate and mometasone furoate. The present invention also relates to pharmaceutical compositions containing a dual PI3K delta and gamma inhibitor and a corticosteroid.

Description

(12) Granted patent speciﬁcaon (19) NZ (11) 766879 (13) B2 (47) Publicaon date: 2021.12.24 (54) METHOD OF TREATMENT AND COMPOSITIONS COMPRISING A DUAL PI3K DELTA-GAMMA KINASE INHIBITOR AND A CORTICOSTEROID (51) Internaonal Patent Classiﬁcaon(s): A61K 45/06 A61K 31/519 A61K 31/56 A61K 31/573 A61P 37/00 A61P 11/06 A61P 11/00 (22) Filing date: (73) Owner(s): 2015.09.03 RHIZEN PHARMACEUTICALS SA (23) Complete speciﬁcaon ﬁling date: (74) Contact: 2015.09.03 Spruson & Ferguson Pty Ltd (62) Divided out of 729419 (72) Inventor(s): KUMAR VENKATA SATYA ANKA, Sw (30) Internaonal Priority Data: aroop IN 4287/CHE/2014 2014.09.03 VISWANADHA, Srikant (57) Abstract: This present disclosure s to a method of treang autoimmune, respiratory and/or inﬂammatory diseases or condions, e.g., , COPD, rheumatoid arthris and thic Pulmonary Fibrosis (IPF). The method comprises administering a dual PI3K delta and gamma inhibitor and a corcosteroid, wherein the corcosteroid is selected from dexamethasone, ﬂucasone, ﬂucasone propionate and mometasone furoate. The t invenon also relates to pharmaceucal composions containing a dual PI3K delta and gamma inhibitor and a corcosteroid. 766879 B2 METHOD OF TREATMENT AND COMPOSITIONS COMPRISING A DUAL PI3K DELTA-GAMMA KINASE INHIBITOR AND A CORTICOSTEROID The present application is a divisional ation of New d Application No. 729419, which is incorporated in its entirety herein by reference.

The present application claims the benefit of Indian Patent Application No. 4287/CHE/2014, filed September 03, 2014, which is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION The present invention relates to a method of treating autoimmune, respiratory and/or inflammatory es and conditions comprising administering to a t in need f a dual PI3K delta/gamma inhibitor and at least one corticosteroid. In preferred embodiments, the method relates to the treatment of psoriasis, rheumatoid arthritis, idiopathic pulmonary fibrosis (IPF), asthma, chronic obstructive pulmonary disease (COPD), and any combination thereof.

BACKGROUND OF THE INVENTION Autoimmune, respiratory and inflammatory diseases such as rheumatoid arthritis (RA), thic pulmonary fibrosis (IPF), psoriasis, systemic lupus erythematosus (SLE), COPD and asthma are chronic and often progressive diseases associated with a dysregulated or an overactive immune , respectively. The causes and the drivers of these diseases remain ill defined. They are typically characterized by complex ar interactions between multiple inflammatory cells of the innate and adaptive immune system. Accordingly, the heterogeneity and complexity of the disease gy of these conditions makes the search for new appropriate cellular targets nging, as it is unclear who in the cellular infiltrate is a primary player of the pathology versus an “innocent” der. Therefore, targeting signalling molecules that are required for the activation of multiple immune cells may be the more likely route to success in combating these chronic, immune cell mediated diseases.

Rheumatoid arthritis (RA) is a progressive, systemic mune disease characterized by chronic inflammation of le joints with associated systemic symptoms such as e. This inflammation causes joint pain, stiffness and swelling, resulting in loss of joint function due to destruction of the bone and cartilage, often leading to progressive WO 35032 lity. Patients with RA also have an increased likelihood of developing other systemic complications such as orosis, anaemia, and others affecting the lungs and skin.

RA is one of the most common forms of autoimmune disease and affects over 21 million people worldwide. Rheumatoid arthritis has a worldwide distribution with an estimated prevalence of 1 to 2%. Prevalence increases with age, approaching 5% in women over the age of 55. The average annual incidence in the United States is about 70 per 100,000 annually. Both incidence and prevalence of rheumatoid arthritis are two to three times greater in women than in men. Although rheumatoid arthritis may present at any age, patients most commonly are first affected in the third to sixth decades. RA is known to impact quality of life, causing not only physical problems but also significant ve impact on quality of life. RA also impacts on the average life expectancy, shortening it by three to seven years.

After 10 years, less than 50% of patients with RA can work or function normally on a day-to- day basis. RA is also been reported to lead to an economic burden on national economies due to hospital ions, health care costs and lost tivity. RA is the cause of over nine million primary care physician visits in the UK annually, representing £833 million in lost tion. It is also ted to have cost the UK economy £55 billion in 2000. In the US, s have estimated that RA costs more to business and industry than any other disease, with 500,000 hospitalizations per year and the burden of illness on the y for arthritis (as a whole) is estimated to be $128 billion.

There are a number of treatments available to manage RA. Some address the signs and symptoms of RA, others aim to modify the course of the disease and positively impact the systemic effects of RA, such as fatigue and anaemia.

The current treatments include, for example, use of: o Biologics: These are genetically-engineered drugs that target specific cell surface markers or messenger nces in the immune system called cytokines, which are produced by cells in order to regulate other cells during an inﬂammatory se. An example of a specific cytokine targeted by biologics is tumor is factor alpha (TNFoc). 0 Traditional disease-modifying anti-rheumatic drugs (DMARDs): These are non- speciﬁc immunosuppressive drugs, which are intended to combat the signs and symptoms of RA as well as slowing down progressive joint destruction. These treatments are often used in combination with one another, or in combination with a biologic agent, to improve patient response WO 2016035032 0 Glucocorticoids (corticosteroids): These are anti-inﬂammatory drugs related to cortisol - a d produced naturally in the body - that work by countering inﬂammation. However, the side-effects of glucocorticoids, which include hyperglycemia, osteoporosis, hypertension, weight gain, cataracts, sleep problems, muscle loss, and susceptibility to ions, limits their use 0 Non-steroidal anti-inﬂammatory drugs (NSAIDs): These manage the signs and symptoms of RA, such as reducing pain, swelling, and inﬂammation, but do not alter the course of the disease or slow the progression ofj oint destruction There are also a number of RA therapies targeting other components of the immune system. These include biologic treatments ing alternative cytokines such as interleukin-6 (IL-6) that help to reduce ation and the progression of RA in the joints and throughout the body.

Asthma is the most common chronic disease among children and also affects millions of adults. Some 235 million people worldwide suffer from this disease. The causes of asthma are not well understood, but effective medicines are available that can treat it, thus largely avoiding the diminished lives, disabilities and death it can bring. Unfortunately, for many people with asthma, particularly the poor, these effective treatments are too costly or not available at all. c obstructive pulmonary disease (COPD) is a highly prevalent condition and a major cause of morbidity and mortality worldwide. As the disease sses, patients with COPD may become prone to frequent bations, resulting in t anxiety, worsening health status, lung function e, and increase in mortality rate. These episodes of worsening atory function lead to increases in health care utilization, hospital admissions and costs. Worse, frequent exacerbations are associated with a faster decline in lung on, y shortening life expectancy.

According to the recommendations of Global Initiative for Chronic Obstructive Lung Disease (GOLD), the first line therapy for COPD are long acting [3- agonists, long acting muscarinic antagonists and tion corticosteroids. However, these drugs reduce the symptoms and exacerbations associated with the disease rather than targeting its molecular and cellular basis. Accordingly, there is still a need for further improvement of COPD therapy.

Phosphoinositide-3 kinase (PI3K) belongs to a class of intracellular lipid kinases that phosphorylate the 3 position yl group of the inositol ring of phosphoinositide lipids (PIs) generating lipid second messengers. While alpha and beta isoforms are ubiquitous in their distribution, expression of delta and gamma is restricted to circulating hematogenous cells and endothelial cells. Unlike PI3K-alpha or beta, mice lacking expression of gamma or delta do not show any e phenotype indicating that targeting of these speciﬁc isoforms would not result in overt toxicity.

Recently, targeted tors of the phosphoinositidekinase (PI3K) pathway have been suggested as immunomodulatory agents. This interest stems from the fact that the PI3K pathway serves multiple functions in immune cell signalling, primarily through the generation of phosphatidylinositol (3,4,5)—trisphosphate , a membrane bound second messenger. PIP3 recruits proteins to the cytoplasmic side of the lipid bilayer, including protein s and GTPases, initiating a complex network of downstream signalling cascades important in the regulation of immune cell adhesion, migration, and ell communication.

The four class I PI3K isoforms differ signiﬁcantly in their tissue distribution.

PI3KOL and PI3KB are ubiquitous and activated downstream of receptor tyrosine s (RTK), whereas PI3K5 and PI3Ky are primarily limited to hematopoietic and endothelial cells, and are activated downstream of RTKs, and G protein coupled receptors (GPCR), respectively. Mouse genetic s have revealed that PI3KOL and PI3KB are essential for normal development, whereas loss of PI3K5 and/or PI3Ky yields viable offspring with selective immune deﬁcits The expression pattern and functions of PI3K5 and PI3Ky have generated much interest in developing PI3K 6/7 inhibitors as agents for many diseases, ing rheumatoid arthritis, ies, asthma, chronic obstructive pulmonary disease and multiple sclerosis h et al., Pharmacol. Ther.,118, 192—205 2008; Marone et al., Biochim.

Biophys. Acta., 1784, 5. 2008, Rommel et al., Nat. Rev. Immunol., 7, 191—201., 2007, Ruckle et al., Nat. Rev. Drug Discov., 5, 82006). Studies using both pharmacologic and genetic methods have shown these two isoforms often demonstrate istic interactions with each other (Konrad ez‘ al., J. Biol. Chem, 283, 33296—33303, 2008, Laffargue et al., Immunity, 16, 441—451, 2002).

In mast cells, for e, PI3K8 is essential for degranulation in response to IgE cross-linking of Fc-receptors (Ali el al., J. Immunol, 180, 2538—2544. 2008), but PI3Ky plays an important role in amplifying the se (Laffargue et al., Immunity, 16, 441—451 2002). Similar effects have been seen in other cellular functions, including lymphocyte homing and the neutrophil respiratory burst where PI3Ky plays a critical role and PI3K5 ampliﬁes each process. The nonredundant but related roles of PI3K6 and PI3Ky have made it difﬁcult to determine which of the two ms (alone or in combination) is best targeted in a particular inﬂammatory disorder. Studies using mice that lack PI3K6 and/or PI3Ky or express kinase-dead ts of PI3K6 and PISKy have been valuable tools in understanding their roles. For e, PI3K6 knockout mice demonstrated diminished neutrophil chemotaxis, diminished antibody tion (both T cell dependent and independent) (Jou et al, Mol.

Cell. Biol., 22, 8580—8591. 2002), and lower numbers of mature B cells (Clayton el al., J.

Exp. Med, 196, 3. 2002, Jou et al, Mol. Cell. Biol., 22, 8580—8591. 2002) and a decrease in their eration in response to anti-IgM (Jou ei al, Mol. Cell Biol, 22, 8580— 8591. 2002). This phenotype was replicated in the PI3K6 kinase-dead variant and with PI3K8 selective inhibitors along with decreased numbers of and proliferation of mast cells, and an attenuated ic response. The PI3Ky knockout contained higher numbers of, but less responsive, neutrophils, lower numbers of and less responsive macrophages and dendritic cells displayed decreased mast cell degranulation ((Laffargue ei al, Immunity, 16, 441—451 2002), a higher ratio of CD4+ to CD8+ T cells), sed thymocyte apoptosis, diminished induction of CXCR3 on activated T cells and decreased cardiac contractility. This latter effect on cardiac tissue was a concern for chronic dosing of patients with PI3Ky inhibitors.

However, this concern was largely ted when the PI3Ky kinase-dead variant (which better mimics inhibition of the kinase rather than loss of the protein) showed similar immune cell phenotypes, but importantly had no cardiac defects. The cardiac effect was later shown to be due to scaffolding effects rather than the catalytic activity of PI3Ky. The dual PI3K5 /PI3Ky knockout was viable but exhibited serious defects in T cell pment and thymocyte survival. The PI3Ky knockout/ PI3K8 kinase-dead combination produced a similar phenotype suggesting that at least within the immune system, the role of PI3K6 is likely only a catalytic one. Interpretation of studies using knockout and kinase—dead mice can be challenging because these models provide only a steady-state picture of the immune system, lack al and dose l, and do not permit a full understanding of how a dynamic immune response will react to reversible inhibition. Selective inhibitors with varying proﬁles (PI3K6, PI3Ky, and PI3K S/y) are necessary for studies of leukocyte signalling in order to assess the relative contributions of each PI3K to immune cell activation. (see Olusegon et al, Chemistry & Biology, 1,123-134 ing the cited nces therein).

Dual inhibition of PI3K 6/3! is strongly implicated as an intervention strategy in allergic and non-allergic ation of the airways and other autoimmune es.

Scientiﬁc evidence for PI3K-6 and y gamma involvement in various cellular processes underlying asthma and COPD stems from tor studies and gene-targeting approaches.

Also, resistance to conventional therapies such as corticosteroids in several COPD patients has been attributed to an up-regulation of the PI3K S/y pathway. Disruption of PI3K 5/7 signalling therefore provides a novel strategy aimed at counteracting the immuno- atory response. Due to the pivotal role played by PI3K 6 and y in mediating inﬂammatory cell functionality such as leukocyte ion and activation, and mast cell degranulation, blocking these isoforms may also be an effective strategy for the treatment of rheumatoid arthritis as well. Given the established criticality of these isoforms in immune surveillance, inhibitors speciﬁcally targeting the delta and gamma isoforms would be expected to attenuate the progression of immune response encountered in airway inﬂammation and toid arthritis. Given the established criticality of these isoforms in immune llance, inhibitors speciﬁcally targeting the 6 and y isoforms would be expected to attenuate the progression of immune response encountered in airway inﬂammation and rheumatoid arthritis (William et al., Chemistry & Biology, 17: 123—134, 2010; and Thompson, et al., Chemistry & Biology, 17 :101-102, 2010)Reviews and studies regarding PI3K and related n kinase pathways have been given by Pixu Liu et. al. e Reviews Drug Discovery, 2009, 8, 4), Nathan T. et. al., Mol Cancer Ther., 2009;8 (1) Jan, 2009), Romina Marone et al., mica et Biophysica Acta., 1784 (2008) 159-185) and B.

Markman et al., Annals of Oncology, Advance Access published August 2009). Similarly reviews and studies regarding role of PI3K 8 and V have been given by William et al., try & Biology, 17:123-134, 2010 and Timothy et al., JMea’. Chem, Web Publication August, 27, 2012. All of these ture disclosures are incorporated herein as reference in their entirety for all purposes.

Recent developed compounds, such as [PI-145 and CAL130 have been ed as dual inhibitors of Pi3K 6/y. IPI-145 is under clinical igation for cancer as well as for asthma. There are currently no reports of CAL-130 being investigated for any clinical purpose.

Additional reference is made herein to International Patent Application Nos. , ﬁled November 3, 2010, and , ﬁled May 4, 2012, US. Patent Application Nos. 12/938,609, ﬁled November 3, 2010, and 13/464,587 ﬁled May 4, 2012 as well to the compounds as disclosed in International Publication Nos. WO 2009/088986, , and , each of which is orated herein by reference in its entirety for all purposes.

Corticosteroids are potent anti-inﬂammatory agents, able to se the number, activity and movement of inﬂammatory cells. Corticosteroids are commonly used to treat a wide range of chronic and acute inﬂammatory conditions including asthma, chronic obstructive pulmonary disease (COPD), allergic rhinitis, rheumatoid arthritis, inﬂammatory bowel disease and autoimmune diseases, Corticosteroids mediate their effects through the orticoid receptor (GR). The binding of corticosteroids to GR induces its nuclear translocation which, in turn, affects a number of downstream pathways via DNA-binding- dependent (e.g. transactivation) and -independent (e. g. transeXpression) mechanisms.

Corticosteroids for treating chronic inﬂammatory conditions in the lung (such as asthma and COPD) are currently administered through inhalation. One of the advantages of employing d corticosteroids (ICS) is the possibility of delivering the drug directly to the site of action, thereby limiting systemic side-effects, and resulting in a more rapid clinical response and higher therapeutic ratio.

Although ICS treatment can afford ant beneﬁts, especially in asthma, it is important to minimize ICS systemic exposure, which leads to the occurrence and severity of unwanted side effects that may be associated with chronic administration. Moreover, the limited duration of action of ICS currently available in the al practice contributes to suboptimal management of the disease. While inhaler technology is an ant point to target the lung, the modulation of the substituents on the corticosteroids lar scaffold is important for the optimization of pharmacokinetic and pharmacodynamic properties in order to decrease oral bioavailability, confine pharmacological activity only in the lung (prodrugs and soft drugs) and increase systemic nce. Moreover, long lasting ICS ty in the lung is highly desirable as once daily administration of ICS would allow the ion of the frequency of stration and, thus, substantially improve t compliance and, as a result, disease ment and control. In sum, there is a pressing medical need for developing ICS with improved pharmacokinetic and pharmacodynamic characteristics.

Glucocorticoids isoxazolidine tives are described, for example, in WO 2006/005611, GB 1,578,446 and in ”Synthesis and topical anti-inﬂammatory activity of some dal [l60t,l70t-d] isoxazolidines”, M.J. Green et al., J. Med. Chem, 25, 1492-1495, 1982, each of which is incorporated herein by reference in their entireties. Additional orticoid isoxazolidine derivatives are also described in WO 2011/029547 and WO 2012/123482.

Despite currently available intervention therapies, autoimmune disorders such as RA, psoriasis and respiratory disorders such as asthma and COPD remains disease classes with a significant unmet medical need.

Accordingly, it is an objective of the present invention to provide methods and pharmaceutical compositions for the ent of atory and/or inflammatory diseases and conditions having enhanced activity. The pharmaceutical itions allow for treating autoimmune, respiratory and inflammatory diseases and conditions with a smaller amount of active compound(s) and/or allow for treating autoimmune, respiratory and inflammatory diseases and conditions in a more efficient way, thereby zing or obviating ly existing adverse effects generally linked to any kind of treatment with an active compound in high doses and/or for a longer period of time.

As described herein, the objective may be ed by combining drugs affecting two diverse yet complimentary pathways, in order to be efficacious at lower doses compared to that of either inhibitor alone. Thus, the present invention provides an effective approach of combining the two different signalling pathways which hold significant therapeutic potential when combined together. In particular the combination is eutically beneficial in ng the required therapeutically effective concentration of either or both the corticosteroid and the dual PI3K deltagamma inhibitor.

SUMMARY OF THE INVENTION The present invention relates to a pharmaceutical composition comprising a PI3K delta and gamma dual inhibitor and at least one osteroid, and to the use of such a pharmaceutical ition for treating autoimmune, respiratory and inflammatory diseases and conditions. [27a] According to a first aspect, the present invention provides use of (i) a dual PI3K delta and gamma inhibitor, and (ii) a osteroid in the manufacture of a medicament for treating an autoimmune, respiratory and/or inflammatory disease or condition, wherein the dual PI3K delta and gamma inhibitor and the corticosteroid are to be administered in a therapeutically effective amount, wherein the corticosteroid is selected from dexamethasone, fluticasone, fluticasone propionate, sone furoate, and pharmaceutically able salts thereof. [27b] ing to a second aspect, the present invention provides a pharmaceutical composition comprising (i) a dual PI3K delta and gamma inhibitor, (ii) a corticosteroid, and (iii) optionally, a pharmaceutically acceptable carrier, glidant, diluent, or excipient, 8a (followed by page 9) wherein the corticosteroid is selected from dexamethasone, fluticasone, asone propionate, and mometasone furoate, and pharmaceutically acceptable salts f. [27c] According to a third aspect, the present invention es the use of a pharmaceutical composition according to the invention in the manufacture of a medicament for the treatment of autoimmune, respiratory and inflammatory diseases and conditions are selected from asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, inflammatory bowel disease, glomerulonephritis, neuroinflammatory diseases, multiple sclerosis, uveitis, sis, tis, vasculitis, dermatitis, osteoarthritis, inflammatory muscle disease, allergic rhinitis, vaginitis, interstitial cystitis, derma, osteoporosis, eczema, allogeneic or xenogeneic transplantation (organ, bone , stem cells and other cells and tissues) graft rejection, graft-versus-host disease, lupus erythematosus, inflammatory disease, type I diabetes, pulmonary fibrosis, dermatomyositis, Sjogren's me, thyroiditis (e.g., oto's and autoimmune thyroiditis), myasthenia gravis, autoimmune hemolytic anemia, cystic fibrosis, Idiopathic pulmonary fibrosis (IPF), chronic relapsing hepatitis, primary biliary cirrhosis, allergic conjunctivitis and atopic dermatitis, and combinations thereof. [27d] According to a fourth aspect, the t invention provides a kit for treating an autoimmune, atory or inflammatory disease or condition, the kit comprising: (i) a dual PI3K delta and gamma tor, and (ii) a corticosteroid, or a pharmaceutically acceptable salt thereof, in a single ceutical composition, (ii) instructions for treating the mune, respiratory or inflammatory disease or condition with the dual PI3K delta and gamma inhibitor and corticosteroid and (iii) a container for placing the pharmaceutical composition or pharmaceutical compositions, wherein the corticosteroid is selected from thasone, fluticasone, fluticasone propionate, and mometasone furoate, and pharmaceutically acceptable salts thereof.

Another embodiment is a pharmaceutical composition comprising a PI3K delta and gamma dual inhibitor and at least one corticosteroid.

Another ment is a method of treating a t suffering from an autoimmune, respiratory and/or inflammatory disease or condition comprising administering to the patient a PI3K delta and gamma dual inhibitor and at least one corticosteroid. In one preferred embodiment, the PI3K delta and gamma dual inhibitor and at least one corticosteroid are administered together in a single pharmaceutical composition. In one WO 35032 preferred embodiment, the disease or condition is idiopathic pulmonary ﬁbrosis (IPF), asthma, rheumatoid arthritis (RA) or COPD.

Yet another embodiment is the use of a combination of a PI3K delta and gamma dual inhibitor and at least one corticosteroid for the treatment in a patient of an autoimmune, respiratory and/or inﬂammatory disease or condition, such as for the treatment of asthma, RA or COPD.

In a preferred embodiment, the PI3K delta and gamma dual inhibitor is a compound of formula A (shown below) or a pharmaceutically acceptable salt thereof.

Formula A Suitable corticosteroids e, but are not limited to dexamethasone, thasone, prednisolone, methyl prednisolone, prednisone, hydrocortisone, ﬂuticasone, triamcinolone, cortisone, rt, deﬂazacort, halopredone acetate, budesonide, beclomethasone dipropionate, hydrocortisone, triamcinolone acetonide, ﬂuocinolone acetonide, ﬂuocinonide, clocortolone pivalate, methylprednisolone aceponate, dexamethasone palmitoate, tipredane, hydrocortisone aceponate, carbate, alclometasone ionate, halometasone, methylprednisolone suleptanate, mometasone furoate, lone, prednisolone farnesylate, ciclesonide, deprodone propionate, fluticasone propionate, halobetasol propionate, ednol etabonate, betamethasone butyrate propionate, lide, prednisone, dexamethasone sodium phosphate, triamcinolone, betamethasone l7-Valerate, betamethasone, betamethasone ionate, hydrocortisone acetate, hydrocortisone sodium succinate, prednisolone sodium phosphate, hydrocortisone probutate, and pharmaceutically acceptable salts thereof.

In a preferred embodiment, the corticosteroids are ed from dexamethasone, betamethasone, prednisolone, methyl prednisolone, prednisone, hydrocortisone, ﬂuticasone, triamcinolone, budesonide, one, and any combination of any of the foregoing.

One embodiment is a pharmaceutical ition comprising a compound of formula A or a pharmaceutically acceptable salt thereof, and a corticosteroid. In one red embodiment, the pharmaceutical composition comprises a therapeutically effective amount of a compound formula A or a ceutically acceptable salt thereof, and a therapeutically effective amount of a corticosteroid (for example, for treating asthma, RA, or COPD).

Another embodiment is a method of treating an autoimmune, respiratory and/or atory disease or condition, such as asthma, RA or COPD, comprising administering to a patient in need thereof a compound of formula A: Formula A or a pharmaceutically acceptable salt thereof and a osteroid. In one preferred embodiment, the compound of formula A or a pharmaceutically acceptable salt thereof and at least one corticosteroid are administered together in a single ceutical composition. In one embodiment, the disease or condition is asthma. In another embodiment, the disease or condition is RA. In yet another ment, the disease or condition is COPD.

Yet another embodiment is a method of treating a patient suffering from an autoimmune, respiratory and/or inﬂammatory e or condition, such as , RA, or COPD, comprising administering to the patient a compound of formula A or a pharmaceutically acceptable salt thereof, and a corticosteroid selected from dexamethasone, betamethasone, prednisolone, methyl prednisolone, prednisone, ortisone, ﬂuticasone, triamcinolone, budesonide or cortisone, and any combination thereof. In one preferred embodiment, the compound of formula A or a pharmaceutically acceptable salt thereof and at least one corticosteroid are administered together in a single pharmaceutical composition.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1A is a bar graph depicting the effect of compound A on the ICso of dexamethasone (Dex) in TGF-Bl treated A549 cells according to the procedure in Example 1.

Figure 1B is a bar graph depicting the effect of compound A on the ICso of dexamethasone (Dex) on 1L-8 concentrations in H202 treated U937 cells according to the procedure in Example 2‘ Figure 2A is a bar graph depicting the effect of compound A on cigarette smoke induced immune cell inﬁltration in BALF of Balb/c mice according to the procedure in Example 3.

Figure 2B is a bar graph depicting the effect of compound A on cytokines in BALF ing to the procedure in Example 3.

Figure 3A is a bar graph ing the effect of compound A and Fluticasone on cigarette smoke d macrophage inﬁltration in BALF of Balb/c mice according to the procedure in e 4.

Figure 3B is a bar graph depicting the effect of combination of compound A and Fluticasone on cigarette smoke induced macrophage infiltration in BALF of Balb/c mice according to the procedure in Example 4.

Figure 4 is a bar graph depicting the IL-8 concentration-dependent inhibitory curve for phils from healthy and COPD patients stimulated with CSE 5% in the presence of Compound A (0.01 nM - 100 uM) or dexamethasone according to the procedure in Example 6.

Figure 5 is a bar graph ing inhibition of duced IL-8 release in neutrophils from COPD patients by addition of a fixed concentration of dexamethasone 1 nM to concentrations of Compound A of O.lnM, lnM, and lOnM according to the procedure in Example 6.

Figure 6 is a bar graph depicting relative MKPl mRNA expression stimulated with CSE 5% alone or in the presence of 10 nM or 100 nM of Compound A according to the procedure in e 7.

Figure 7 is a bar graph depicting relative PI3Ky mRNA expression stimulated with CSE 5% alone or in the ce of 10 nM or 100 nM of Compound A according to the procedure in e 7.

Figure 8 is a bar graph depicting PIP3 production in the presence of CSE 5% alone, CSE5% and 10 nM of Compound A, or 10 nM of Compound A alone.

DETAILED DESCRIPTION OF THE INVENTION In one aspect, the method of combining a dual PI3K delta and gamma inhibitor (such as a compound of formula A, or a pharmaceutically acceptable salt thereof) with a corticosteroid, as described in any of the embodiments herein, ts an activity (i.e., a istic activity) which is signiﬁcantly higher than the ty expected based on the individual activities of each of the dual PI3K delta and gamma inhibitor or the corticosteroid alone.

In another aspect, the method of combining a dual PI3K delta and gamma inhibitor (such as a compound of formula A, or a pharmaceutically acceptable salt thereof) with a corticosteroid exhibits an activity even when the osteroid alone is insensitive as a single agent.

Thus, the methods described herein allow for treating autoimmune, respiratory and atory diseases and conditions with a smaller amount of active compound(s) and/or allow for treating autoimmune, atory and atory diseases and conditions for a longer period of time in a more efﬁcient way.

Another embodiment is a pharmaceutical composition comprising a dual PI3K delta and gamma inhibitor (such as a compound of formula A, or a pharmaceutically acceptable salt f) with a corticosteroid, for use in the treatment of an mune, respiratory and/or inﬂammatory disease or condition.

Yet another embodiment is a method of treating an autoimmune, respiratory and/or inﬂammatory disease or condition comprising administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition according to the present invention.

Yet another embodiment is the use of a pharmaceutical composition according to any of the embodiments described herein for making a medicament useful for treating an mune, atory and/or inﬂammatory disease or condition.

In the pharmaceutical compositions described herein, the PI3K delta and gamma dual inhibitor (such as a compound of formula A, or a pharmaceutically acceptable salt thereof) may be in a form selected from solvates, hydrates and/or salts with cologically able acids or bases.

In the pharmaceutical compositions described herein, the corticosteroid may be in a form selected from solvates, hydrates or salts with pharmacologically acceptable acids or bases. 2015/056720 Yet another embodiment is a method of treating an immune system-related disease (e.g., an autoimmune disease), a disease or er involving inﬂammation (e.g., asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, inﬂammatory bowel disease, glomerulonephritis, neuroinﬂammatory diseases, multiple sclerosis, uveitis and disorders of the immune system), cancer or other proliferative disease, a c disease or er, or a renal disease or disorder. The method includes administering an effective amount of one or more compositions of the present invention.

Examples of immune disorders which can be d by the methods and compositions described herein include, but are not limited to, psoriasis, rheumatoid arthritis, vasculitis, inﬂammatory bowel disease, dermatitis, osteoarthritis, , inﬂammatory muscle e, allergic rhinitis, vaginitis, interstitial cystitis, scleroderma, osteoporosis, eczema, allogeneic or xenogeneic transplantation (organ, bone marrow, stem cells and other cells and tissues) graft rejection, graft-versus-host disease, lupus erythematosus, inﬂammatory disease, type I diabetes, pulmonary ﬁbrosis, dermatomyositis, Sjogren's syndrome, thyroiditis (e.g., Hashimoto's and autoimmune thyroiditis), myasthenia gravis, mune hemolytic anemia, multiple sclerosis, cystic fibrosis, thic pulmonary ﬁbrosis (IPF), c ing hepatitis, primary biliary cirrhosis, allergic conjunctivitis and atopic dermatitis.

Pharmaceutically acceptable salts, as described herein, include salts derived from inorganic bases such as Li, Na, K, Ca, Mg, Fe, Cu, Zn, and Mn, salts of organic bases such as N,N'-diacetylethylenediamine, glucamine, triethylamine, choline, hydroxide, dicyclohexylamine, metformin, benzylamine, trialkylamine, and thiamine, salts of chiral bases such as alkylphenylamine, glycinol, and phenyl glycinol, salts of natural amino acids such as glycine, e, valine, leucine, isoleucine, cine, tyrosine, cystine, cysteine, methionine, proline, hydroxy proline, histidine, omithine, lysine, arginine, and serine, quaternary um salts of the compounds of invention with alkyl halides, alkyl sulphates such as MeI (methyl iodide) and (Me)2SO4, salts of non-natural amino acids such as D- isomers or substituted amino acids, salts of guanidine; and salts of substituted guanidine n the tuents are selected from nitro, amino, alkyl, alkenyl, alkynyl, ammonium or substituted ammonium salts and aluminum salts. Salts may include acid addition salts where appropriate which are sulphates, nitrates, phosphates, perchlorates, s, alides, acetates, tartrates, maleates, citrates, fumarates, succinates, palmoates, methanesulphonates, benzoates, salicylates, benzenesulfonates, ates, glycerophosphates, and ketoglutarates.

When ranges are used herein, all combinations and subcombinations of ranges and speciﬁc embodiments therein are intended to be included. The term "about" when referring to a number or a numerical range means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error), and thus the number or numerical range may vary from, for e, n 1% and % of the stated number or numerical range. The term "comprising" (and related terms such as "comprise" or "comprises" or "having" or "including") includes those embodiments, for example, an embodiment of any composition of matter, composition, method, or process, or the like, that "consist of’ or "consist essentially of’ the described features.

The following abbreviations and terms have the indicated meanings throughout: P13 -K = Phosphoinositide 3-kinase; P1 = phosphatidylinositol.

Abbreviations used herein have their conventional meaning within the chemical and biological arts, unless otherwise indicated.

The term "effective amount" or "therapeutically effective amount" refers to that amount of a compound bed herein that is sufﬁcient to effect the intended ation including, but not d to, disease treatment, as deﬁned below. The therapeutically effective amount may vary depending upon the intended application (in vitro or in vivo), or the subject and disease condition being treated, e.g., the weight and age of the subject, the ty of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art. The term also applies to a dose that will induce a particular response in target cells, e.g., reduction of platelet adhesion and/or cell migration. The speciﬁc dose will vary depending on the particular compounds chosen, the dosing regimen to be followed, whether it is administered in combination with other compounds, timing of administration, the tissue to which it is administered, and the physical delivery system in which it is carried.

As used herein, the terms ment" and "treating" refer to an approach for ing beneﬁcial or desired results including, but not limited to, eutic beneﬁt and/or a prophylactic beneﬁt. By therapeutic beneﬁt is meant eradication or amelioration of the underlying disorder being treated. Also, a therapeutic beneﬁt is achieved with the eradication or amelioration of one or more of the physiological ms associated with the underlying er such that an improvement is ed in the patient, notwithstanding that the patient may still be ed with the underlying disorder. For prophylactic beneﬁt, the itions may be administered to a patient at risk of ping a particular disease, or to a patient reporting one or more of the physiological symptoms of a disease, even though a diagnosis of this disease may not have been made.

A "therapeutic effect," as that term is used herein encompasses a therapeutic beneﬁt and/or a prophylactic benefit as described above. A lactic effect includes delaying or eliminating the appearance of a disease or ion, delaying or eliminating the onset of ms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof.

The term "subject" or “patient” refers to an animal, such as a mammal, for example a human. The methods described herein can be useful in both human therapeutics and veterinary applications. In some embodiments, the patient is a mammal, and in some embodiments, the patient is human. For nary purposes, the term “subject” and nt” include, but are not limited to, farm animals including cows, sheep, pigs, horses, and goats; companion animals such as dogs and cats; exotic and/or zoo animals; laboratory animals including mice, rats, rabbits, guinea pigs, and hamsters; and y such as chickens, turkeys, ducks, and geese.

The term "selective inhibition" or "selectively inhibit" as applied to a biologically active agent refers to the agent's ability to selectively reduce the target signaling activity as compared to off—target signaling activity, via direct or indirect interaction with the target.

As used herein, the term “dual PI3—kinase Delta (6) and Gamma (y) tor" generally refers to a compound that inhibits the activity of both the PI3-kinase 6 and y isozyme more effectively than other isozymes of the PI3K family. A PI3-kinase 6 and 7 dual inhibitor compound is therefore more ive for PI3 -kinase 6 and 7 than conventional PI3K inhibitors such as CAL-130, wortmannin and LY294002, which are "nonselective PI3K inhibitors." es of “dual PI3-kinase Delta (6) and Gamma (7) inhibitor" e, but are not limited to, compounds such as IPI—l45, and the compounds disclosed in International Patent Application Nos. , ﬁled er 3, 2010, and PCT/U82012/36594, ﬁled May 4, 2012; US. Patent Application Nos. 12/938,609, ﬁled November 3, 2010, and 13/464,587 ﬁled May 4, 2012 and to compounds disclosed in International Publication Nos. , , and , each of which is orated herein by reference in its entirety for all purposes.

WO 35032 For instance, the Dual PI3-kinase 6 and y selective inhibitor may refer to a nd that exhibits a 50% inhibitory tration (ICso) with respect to the delta and gamma type I PI3-kinase that is at least 10—fold, at least 20-fold, at least 50-fold, or at least lOO-fold lower than the inhibitor's ICso with respect to the other types of P13 kinases (i.e., alpha and beta).

Inhibition of PI3-kinase 5 and y may be of therapeutic beneﬁt in treatment of various ions, e.g., conditions characterized by an inﬂammatory response including but not d to autoimmune diseases, allergic diseases, and arthritic diseases. Importantly, inhibition of PI3-kinase 6 and y function does not appear to affect biological functions such as viability and fertility.

"Inﬂammatory response" as used herein is characterized by redness, heat, swelling and pain (i.e., inﬂammation) and typically involves tissue injury or destruction. An inﬂammatory response is y a localized, protective response elicited by injury or destruction of tissues, which serves to destroy, dilute or wall off (sequester) both the injurious agent and the injured tissue. Inﬂammatory responses are y ated with the inﬂux of leukocytes and/or leukocyte (e.g., neutrophil) chemotaxis. Inﬂammatory responses may result from infection with pathogenic organisms and viruses, non-infectious means such as trauma or reperfusion following myocardial tion or stroke, immune responses to foreign antigens, and autoimmune diseases. Inﬂammatory responses amenable to ent with the s and compounds according to the invention encompass conditions ated with reactions of the c defence system as well as conditions associated with reactions of the eciflc defence system.

The therapeutic methods of the invention include methods for the treatment of conditions associated with inﬂammatory cell tion. "Inﬂammatory cell activation" refers to the induction by a stimulus (including, but not limited to, cytokines, ns or auto- antibodies) of a proliferative cellular response, the production of soluble mediators (including but not limited to cytokines, oxygen radicals, enzymes, prostanoids, or vasoactive amines), or cell surface expression of new or increased numbers of mediators (including, but not limited to, major histocompatibility antigens or cell adhesion molecules) in inﬂammatory cells (including, but not limited to, monocytes, macrophages, T lymphocytes, B lymphocytes, granulocytes (polymorphonuclear leukocytes ing neutrophils, basophils, and eosinophils) mast cells, dendritic cells, Langerhans cells, and endothelial cells). It will be appreciated by persons skilled in the art that the activation of one or a combination of these phenotypes in these cells can contribute to the initiation, perpetuation, or exacerbation of an inﬂammatory condition.

"Autoimmune e" as used herein refers to any group of disorders in which tissue injury is associated with humoral or cell-mediated responses to the body's own constituents.

"Transplant rejection" as used herein refers to an immune response directed against grafted tissue (including organs or cells (e.g., bone marrow), characterized by a loss of function of the grafted and surrounding tissues, pain, swelling, leukocytosis, and thrombocytopenia). gic disease" as used herein refers to any symptoms, tissue damage, or loss of tissue function resulting from allergy.

"Arthritic e" as used herein refers to any disease that is characterized by inﬂammatory lesions of the joints attributable to a variety of etiologies.

"Dermatitis" as used herein refers to any of a large family of diseases of the skin that are characterized by inﬂammation of the skin attributable to a variety of etiologies.

One embodiment is a pharmaceutical composition sing a dual PI3K delta and gamma inhibitor (such as a compound of formula A, or a pharmaceutically able salt thereof) and at least one osteroid and ally one or more pharmaceutically acceptable rs or excipients.

In one embodiment, the pharmaceutical composition includes a therapeutically effective amount of a dual PI3K delta and gamma inhibitor (such as a compound of formula A, or a pharmaceutically acceptable salt thereof) and at least one corticosteroid, and optionally one or more pharmaceutically acceptable carriers or excipients..

The pharmaceutical composition may include one or more additional active ingredients as described herein.

The pharmaceutical carriers and/or excipients may be selected from diluents, ﬁllers, salts, disintegrants, binders, lubricants, glidants, wetting agents, controlled release matrices, colorants, ﬂavorings, buffers, stabilizers, lizers, and combinations thereof.

The pharmaceutical compositions of the present invention can be administered alone or in combination with one or more other active . Where d, the subject compounds and other agent(s) may be mixed into a preparation or both components may be formulated into te ations to use them in combination separately or at the same time.

The dual PI3K delta and gamma inhibitor and the corticosteroid can be administered together or in a tial manner with one or more other active agents. Where desired, the subject compounds and other agent(s) may be co-administered or both components may be administered in a sequence to use them as a combination.

The compounds and pharmaceutical compositions of the present invention can be administered by any route that enables delivery of the compounds to the site of , such as orally, intranasally, topically (e.g., transdermally), intraduodenally, parenterally (including intravenously, intraarterially, intramuscularally, intravascularally, intraperitoneally or by ion or infusion), intradermally, by ammary, intrathecally, cularly, retrobulbarly, intrapulmonary (e.g., aerosolized drugs) or subcutaneously (including depot administration for long term release e.g., embedded-under the-splenic capsule, brain, or in the cornea), sublingually, anally, rectally, vaginally, or by surgical implantation (e.g., embedded under the splenic capsule, brain, or in the cornea).

The compositions can be administered in solid, semi-solid, liquid or gaseous form, or may be in dried powder, such as lyophilized form. The ceutical compositions can be packaged in forms convenient for delivery, including, for example, solid dosage forms such as capsules, sachets, cachets, gelatins, papers, tablets, suppositories, pellets, pills, troches, and lozenges. The type of packaging will lly depend on the desired route of administration. Implantable sustained release formulations are also plated, as are transdermal formulations.

The dosing frequency of the nds may vary. For example, a dual PI3K delta and gamma inhibitor may be administered at a frequency ranging from twice daily to once every three weeks. The corticosteroid may be stered at a ncy ranging from twice daily to once every three weeks.

The amount of the compound to be administered is dependent on the mammal being treated, the severity of the disorder or condition, the rate of administration, the ition of the compound and the discretion of the prescribing physician. However, an effective dosage is in the range of about 0.001 to about 100 mg per kg body weight per day, preferably about 1 to about 35 mg/kg/day, in single or divided doses. For a 70 kg human, this would amount to about 0.05 to 7 g/day, preferably about 0.05 to about 2.5 g/day An effective amount of a compound of the invention may be administered in either single or multiple doses (e.g., twice or three times a day).

In one embodiment, the pharmaceutical compositions described herein comprise from about 0.001 mg to about 1000 mg, such as from about 0.01 mg to about 500 mg or from about 0.010 mg to about 250 mg or from about 0.030 mg to about 125 mg of a dual PI3K delta and gamma inhibitor (such as a compound of formula A, or a pharmaceutically acceptable salt thereof) and/or from about 0.001 mg to about 1000 mg, such as from about 0.01 mg to about 500 mg or from about 0.010 mg to about 250 mg or from about 0.010 mg to about 125 mg or from about 0.030 mg to about 50 mg of at least one corticosteroid.

In one ment, the pharmaceutical compositions described herein comprise the dual PI3K delta and gamma inhibitor and the corticosteroid in a ratio of between about 100:1 and about 1:100 by weight, such as between about 50: 1 and about 1: 50 by weight or between about 1: 10 and about 10: l by weight, or between about 1: 5 and about : 1 by .

The term "co-administration, administered in combination with," and their grammatical equivalents, as used herein, encompasses administration of two or more agents (such as the dual PI3K delta and gamma inhibitor and the corticosteroid) to an animal so that both agents and/or their lites are t in the animal at the same time. Co- administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or stration in a composition in which both agents are present.

The pharmaceutical compositions bed herein may contain one or more osteroids selected form dexamethasone, betamethasone, prednisolone, methyl prednisolone, prednisone, hydrocortisone, ﬂuticasone, triamcinolone budesonide or cortisone prednisolone, methylprednisolone, naﬂocort, deﬂazacort, halopredone acetate, budesonide, beclomethasone dipropionate, hydrocortisone, triamcinolone acetonide, ﬂuocinolone acetonide, ﬂuocinonide, clocortolone pivalate, methylprednisolone aceponate, dexamethasone oate, tipredane, hydrocortisone aceponate, prednicarbate, alclometasone dipropionate, halometasone, methylprednisolone suleptanate, sone furoate, rimexolone, prednisolone farnesylate, ciclesonide, deprodone propionate, ﬂuticasone propionate, halobetasol propionate, loteprednol etabonate, betamethasone butyrate propionate, ﬂunisolide, prednisone, thasone sodium phosphate, triamcinolone, betamethasone l7-valerate, betamethasone, betamethasone dipropionate, hydrocortisone acetate, hydrocortisone sodium succinate, solone sodium phosphate and hydrocortisone probutate, and any combination of any of the foregoing.

In certain embodiments, the corticosteroid is ed from dexamethasone, betamethasone, prednisolone, methyl solone, prednisone, hydrocortisone, ﬂuticasone, triamcinolone, budesonide or cortisone, and any combination thereof.

One particular embodiment of the present invention relates to pharmaceutical itions n the corticosteroid is ﬂuticasone.

Another particular embodiment of the present invention relates to pharmaceutical compositions wherein the corticosteroid is budesonide.

Yet another ular embodiment of the present invention relates to pharmaceutical compositions wherein the corticosteroid is prednisolone.

Yet another particular embodiment of the present invention relates to pharmaceutical compositions wherein the corticosteroid is thasone.

A further ment of the present ion s to a method of ng an indication selected from atory diseases and conditions such as diseases of the airways and lungs which are accompanied by increased or altered production of mucus and/or inﬂammatory and/or obstructive diseases of the airways such as acute bronchitis, chronic bronchitis, chronic obstructive bronchitis (COPD), cough, pulmonary emphysema, allergic or non—allergic rhinitis or sinusitis, chronic sinusitis or rhinitis, nasal polyposis, chronic rhinosinusitis, acute rhinosinusitis, asthma, allergic bronchitis, alveolitis, Farmer's disease, hyperreactive airways, bronchitis or pneumonitis caused by infection, eg. by bacteria or viruses or helminthes or fungi or protozoons or other pathogens, pediatric asthma, bronchiectasis, pulmonary ﬁbrosis, adult respiratory distress me, bronchial and pulmonary edema, bronchitis or pneumonitis or interstitial pneumonitis caused by different origins, e.g. aspiration, tion of toxic gases, vapors, bronchitis or nitis or interstitial nitis caused by heart e, X-rays, radiation, chemotherapy, bronchitis or pneumonitis or interstitial pneumonitis associated with collagenosis, eg. lupus erythematodes, systemic scleroderrna, lung s, idiopathic pulmonary lung fibrosis (IPF), interstitial lung diseases or interstitial pneumonitis of different origin, ing asbestosis, silicosis, M. Boeck or sarcoidosis, granulomatosis, cystic fibrosis or mucoviscidosis, or a-l- antitrypsin deﬁciency, or selected from inﬂammatory diseases and conditions such as inﬂammatory diseases of the gastrointestinal tract of various origins such as inﬂammatory pseudopolyps, Crohn's disease, ulcerative colitis, inﬂammatory diseases of the joints, such as WO 2016035032 rheumatoid arthritis, or ic inﬂammatory diseases of the sopharynx, skin or the eyes, such as atopic dermatitis, seasonal and perenial, chronic uritcaria, hives of unknown cause and allergic conjunctivitis; and in particular selected from asthma, allergic and non- allergic rhinitis, COPD and atopic dermatitis, comprising administering a therapeutically effective amount of a pharmaceutical composition ing to the present invention to a patient in need thereof.

A further embodiment of the present invention relates to the use of a pharmaceutical composition according to the present invention for making a medicament for treating respiratory and/or inﬂammatory diseases and conditions, particularly wherein the respiratory and/or inﬂammatory diseases or conditions are selected from asthma, allergic and non-allergic rhinitis, COPD and atopic dermatitis.

A further embodiment of the present invention s to a pharmaceutical ition according to any embodiment herein, for use in the treatment of respiratory and atory diseases and conditions, particularly wherein the atory and inﬂammatory diseases or conditions are selected from asthma, allergic and non-allergic rhinitis, COPD and atopic dermatitis.

The present invention is now further illustrated by means of the following, non-limiting, examples.

EXAMPLES Provided below are rative es of the ation of a PI3K delta and gamma dual inhibitor and a osteroid.

Example 1: TGF-Bl induced Corticosteroid Insensitivity in A549 Cells Test Procedure A549 cells were trypsinized and 2*104 cells per well were seeded in a 96-well plate and incubated at 37° C and 5% C02.

Media was removed and 100 pl of serum free media with 0.1 uM of Compound A was added and incubated for 30 min. 50 ul of 3X TGF-Bl in F12K with 0.5% BSA was added such that the ﬁnal concentration was 400 pM and incubated at 37° C and 5% CO2 for 4 h. 50 ul of 4X of desired concentrations of dexamethasone (Dex) was added and incubated for 45 min at 37°C and 5% CO2. 50 pl of 5X concentration of TNF-d was added such that the ﬁnal concentration was 1 ng/ml to induce IL-8 and incubated for 24 h.

Supernatant was ted and IL-8 was estimated by ELISA.

Cytokine Assay IL-8 strips were plated with fresh or thawed supernatants and incubated at room temperature for 2 h or overnight at 4° C.

Contents were discarded and strips were washed with 200 pl of wash buffer per well for 15 s for a total of 5 times.

Strips were blotted dry and 100 pl per well of IX detection antibody was added and incubated at room temperature for 1 h, Contents were discarded and strips were washed with 200 pl of wash buffer per well for 15 s for a total of 5 times.

Strips were blotted dry and 100 pl per well of IX Avidin-HRP dy was added and ted at room temperature for 30 mini [1 l 1] Contents were discarded and the strips were washed with 200 p1 per well of wash buffer for 15 s for a total of 5 times. 100 pl per well of TMB substrate were added and incubated at room temperature for 5-15 min.

Reaction was stopped by adding 50 pl per well of 2N H2SO4.

Absorbance was read on a plate reader at A450 nm and A570 nm. % inhibition for Blank subtracted absorbance values were ined based on the control wells. Data was plotted using GraphPad Prism (Version 5.02).

Results The results are depicted in Figure 1A. Compound A (de A) sed the ICso of dexamethasone for IL-8 concentrations in TGF-Bl treated A549 cells indicating signiﬁcant potentiation of thasone activity.

Example 2: H202 Induced Corticosteroid Insensitivity in U937 cells Test Procedure U937 cells were maintained in RPMI—l640 with 15 mM glutamine. 6* 106 cells were taken in T-25 ﬂask with 12 ml of fresh medium and treated with 1 pM of Compound A and incubated at 370 C and 5% C02 for 30 min.

H202 was added at a ﬁnal concentration of 200 pM to the above cells and ted for 2 h.

Cells were pelleted and ended in serum free media and seeded on to a 96-well plate at O. 15*106 cells per well in 100 pl. 50 pl of 3X Dexamethasone at desired concentrations was added and incubated for 45 min. 50 pl of 4X concentration of TNF-u was added such that the ﬁnal concentration was 10 ng/ml, to induce 1L-8 and incubated for 18 h.

Supernatant was collected and lL-8 was estimated by ELISA.

Cytokine Assay IL-8 strips were plated with fresh or thawed supernatants and incubated at room temperature for 2 h or overnight at 4°C.

Contents were discarded and strips were washed with 200 pl of wash buffer per well for 15s for a total of 5 times.

Strips were blotted dry and 100 pl per well of IX detection antibody was added and incubated at room temperature for 1 h.

Strips were blotted dry and 100 pl per well of IX Avidin—HRP antibody was added and incubated at room temperature for 30 min.

Contents were discarded and the strips were washed with 200 pl per well of wash buffer for 15 s for a total of 5 times. 100 pl per well of TMB ate were added and incubated at room temperature for 5-15 min. on was stopped by adding 50 pl per well of 2N H2SO4.

Absorbance was read on a plate reader at A450 nm and A570 nm.

Results As ed in Figure 1B, Compound A (de A) decreased the ICso of dexamethasone (Dex) on IL-8 concentrations in H202 treated U937 cells indicating signiﬁcant potentiation of dexamethasone activity.

Example 3: Chronic Cigarette Smoke Induced Cell Infiltration in Male Balb/c mice Animals were atized for seven days prior to the start of the experiment.

Animals were randomly distributed to s groups based on their body weights. Mice were exposed to the mainstream smoke of 2 ttes from day l to day 11. Exposure to the smoke of each cigarette lasted for 10 min (each cigarette was completely burned in the ﬁrst two minutes, followed by an air ﬂow with animal ator) and were exposed for the next min with fresh room air. After every second cigarette an additional break of 20 min with exposure to fresh room air was conducted. Control animals were exposed to the room air chamber. Test compound was administered by the intranasal route as suspension from day 12 to day 14 before 30 mins whole body smoke exposure. Mice were exposed to the mainstream smoke of l tte from day 12 to day 14. On day 15, 24 hours after the last cigarette smoke (CS) re animals were exsanguinated under anaesthesia, and the trachea was cannulated and the lungs were lavaged with 0.5 ml aliquots of heparinised PBS (1 unit/ml) four times through tracheal a (total volume 2 ml). Bronchioalveolar (BAL) collected was stored at 2-8 °C until assayed for total cell and differential leukocyte count. BAL ﬂuid was centrifuged (500><g for 10 min) and the resulting cell pellet was resuspended in 0.5 ml of heparinised saline. The total number of white blood cells was determined in BAL ﬂuid and blood using a blood cell counter and adjusted to l><lO6 l. Differential cell count was calculated manually. Forty microliters of the cell suspension was centrifuged using cytospin 3 to prepare a cell smear. The cell smear was stained with a blood staining on for differentiation and microscopically observed by identifying each cell according to its morphological characteristics. The number of each cell type among 300 white blood cells in the cell smear was determined and expressed as a percentage, and the number of neutrophils and macrophages in each BAL ﬂuid were calculated. In addition BAL supernatant were analysed for various cytokinines using ELISA assay.

The results are shown in Table l and s 2A and 2B.

All animals survived to the scheduled termination. Compound A showed signiﬁcant beneﬁcial therapeutic effect in the established murine chronic COPD model as determined by evaluation of cell count in BAL. Macrophage inﬁltration in BAL ﬂuid with treated animals differed signiﬁcantly from disease controls with signiﬁcant reductions (toward normal) of BAL cell count seen in mice treated with Compound A (0.003-3 mg/kg) in a dose dependent manner. Macrophage count was cantly reduced toward normal for mice given 0.003-3 mg/kg Compound A. The percent inhibitions of cytokines are given in Table l.

WO 2016035032 Table 1 Cytokines in BALF (% inhibition) Dose (mg/kg) 0 003 0.03 0.3 IL-6 44% 99% TNFot 40% 99% IL-23 0% 100% IFNy 25% 90% Example 4: al of Corticosteroid Insensitivity in Chronic Cigarette Smoke Induced Cell Infiltration in Male Balb/c Mice Animals were acclimatized for seven days prior to the start of the experiment.

Animals were randomly distributed to various groups based on their body weights. Mice were exposed to the mainstream smoke of 2 cigarettes from day 1 to day 11. Exposure to the smoke of each cigarette lasted for 10 min (each cigarette was completely burned in the ﬁrst two minutes, followed by an air ﬂow with animal ventilator) and were exposed for the next min with fresh room air. After every second tte an additional break of 20 min with exposure to fresh room air was conducted. Control animals were exposed to the room air chamber. Corticosteroid, ﬂuticasone was administered by intranasal route from day 6 to day 11 before 30 mins whole body smoke exposure. Mice were exposed to the mainstream smoke of l cigarette from day 12 to day 14. Test compound was administered by the intranasal route as suspension from day 12 to day 14 before 30 mins whole body smoke exposure. On day 15, 24 hours after the last cigarette smoke (CS) exposure animals were uinated under anaesthesia, and the trachea was cannulated and the lungs were lavaged with 0.5 ml aliquots of heparinised PBS (1 l) four times h tracheal cannula (total volume 2 ml).

Bronchioalveolar (BAL) collected was stored at 2-8 °C until assayed for total cell and differential leukocyte count. BAL ﬂuid was centrifuged (500>< g for 10 min) and the resulting cell pellet was resuspended in 0.5 ml of heparinised saline. The total number of white blood cells was determined in BAL ﬂuid and blood using a blood cell counter and adjusted to 1X106 cell/ml. Differential cell count was ated manually. Forty microliters of the cell suspension was fuged using cytospin 3 to prepare a cell smear. The cell smear was stained with a blood staining on for differentiation and microscopically observed by identifying each cell according to its morphological characteristics. The number of each cell WO 2016035032 type among 300 white blood cells in the cell smear was determined and expressed as a percentage, and the number of neutrophils and hages in each BAL ﬂuid were calculated. In addition BAL supernatant were analysed for various cytokines using ELISA assay.

In ation with ﬂuticasone (FLT), Compound A showed signiﬁcant beneficial therapeutic effect and reversal of corticosteroid insensitivity by showing a synergistic effect on macrophage ation. The EDso of the combination was 0.021 mg/kg in the established murine chronic COPD model as determined by evaluation of cell count in BAL compared to an EDso of 0.093 mg/kg of Compound A alone. The results are also shown in Figures 3A and 3B.

Example 5: General Description related to Patient Identiﬁcation, Isolation of Neutrophils and Preparation of Cigarette Smoke Extract (CSE) for in-vitro g of Compound A A. Patient ion Healthy subjects and COPD patients were included for leukocyte experiments.

Pulmonary function tests (forced etry) and arterial blood gas measurements were performed during the days prior to sampling. According to their spirometry results and smoking habits, patients were classiﬁed into two groups: A) Healthy subjects, patients with normal lung function and who did not smoke, B) COPD, patients who had smoked more than pack-years and with airﬂow obstruction evidenced by a forced expiratory volume in l s (FEVl) of <80% predicted and an FEVl forced vital capacity (FVC) ratio of <70%.

Clinical teristics of the patients are provided in Table 2.

Table 2: Clinical features. COPD: chronic obstructive pulmonary disease; FEVl: forced tory volume in one second, FVC: forced vital capacity, r = 1 year smoking 20 cigarettes-day. Data are mean :: SE.

Table 2 Healthy COPD n=7 = e r 66.1i6 65.1:14 Sex,l\/I/F 6/2 Tobacco consumotion, oack- r 35,2: FEVl, % ored FVC, % ored FEVl/FVC % GOLD 1 mild atients, no. __-l_—_ -]—__—_ -I!-I_____ -—__— Receivin- theoh ilines no ____ in- .“- Receivin- anticholiner-ics no “— Peripheral neutrophils and monocytes as well as whole blood were obtained from 8 patients with COPD, deﬁned according to GOLD guidelines and 7 healthy subjects. ts were aged 65.1 i14 years, FEVl 58.2 i 3 % predicted. All patients were current smokers. There were no exacerbations of the disease within 2 weeks prior to taking blood samples. 7 age-matched non-smoking control subjects with normal lung function (age 66.1 i 6 years old, FEVl 98 i 3 % predicted) who did not have any respiratory disease, were also recruited as normal controls, respectively. Routine lung on tests were performed to te forced vital capacity (FVC), forced expiratory volume in 1 s (FEVl) and FEVl/FVC ratio using a Vitalograph® alpha III eter (Vitalograph, Maids Moreton, UK). This project was approved by the local ethics committee of General University Hospital, Valencia, Spain, and written ed consent was taken from each patient or volunteer before starting blood ng and lung function testing.

B. Isolation of Human Neutrophils Neutrophils were isolated from peripheral venous blood by standard laboratory procedures. In brief, peripheral venous blood was mixed with dextran 500 at 3% (in 0.9% ) in a tion of 2:1. This mixture was incubated at room temperature for min until erythrocytes were nted. The upper phase was carefully collected and added on Ficoll-Paque Histopaque 1077 (Amershan Pharrnacia h, Barcelona, Espaﬁa) density gradient in a proportion of 3:1. The two phases generated were centrifuged at 150 g, 4°C for 30 min. Thus, the pellet obtained (which is consisted a mixture of neutrophils and low proportion of residual erythrocytes and traces of eosinophils and basophils) was ended in an erythrocyte lysis buffer (Biolegend, UK) for 5 min in ice. Cell suspension was washed two times with phosphate buffer (PBS). The preparations were >97% pure in neutrophils as assessed by Giemsa staining, and had a viability of >99%, measured by trypan blue exclusion. Neither purity nor viability was affected in the study’s different experimental conditions.

WO 2016035032 C. Preparation of Cigarette Smoke Extract solutions CSE was prepared as follows: Brieﬂy, the smoke of a research cigarette (2R4F, Tobacco Health Research, University of Kentucky, KY, USA) was generated by a respiratory pump (Apparatus Rodent Respirator 680; Harvard, Germany) through a pufﬁng mechanism related to the human smoking pattern (3 puff/min, 1 puff 35 ml, each puff of 2 s duration with 0.5 cm above the ﬁlter) and was bubbled into a ﬂask containing 25 ml of pre- warmed (37°C) Roswell Park Memorial ute (RPMI)-164O culture medium. The CSE solution was sterilized by ﬁltration through a 0.22-um ose e izing system (Corning, NY). The resultant CSE on was considered to be 100% CSE and was used for experiments within 30 min of preparation. CSE 10% corresponds approximately to the exposure associated with smoking two packs per day. The quality of the prepared CSE solution was assessed based on the absorbance at 320 nm, which is the speciﬁc absorption wavelength of peroxynitrite. Stock solutions with an absorbance value of 3.0 i 0.1 were used.

To test for cytotoxicity from CSE, isolated neutrophils were treated with CSE concentrations of up to 5% for 24. No signiﬁcant difference in the lactate dehydrogenase supernatant level (lactate dehydrogenase cytotoxicity assay; Cayman, Spain) was ed in comparison with the l group (data not .

Example 6 Assay: Effect of Compound A, dexamethasone and combination thereof on secretion of inﬂammatory marker IL-8 induced by CSE in peripheral blood neutrophils from healthy non-smokers and COPD smoker patients. ed human phils from healthy volunteers and COPD patients were incubated with Compound A (0.01nM-100uM) and Dexamethasone (0.1nM-1uM) or vehicle for 30 minutes before tion with or without CSE 5% for 6 hours in standard cell culture conditions (37°C and 5% C02). Supernatants were collected to measure different inﬂammatory markers.

IL-8 was measured by ELISA using a commercially ble kit.

Experiments were done in triplicate in almost three patients per experimental condition.

Neutrophils from healthy and COPD patients were stimulated with CSE 5% in the presence of Compound A (0.01nM - 100uM) or thasone (DEX; 0.1nM-1uM) for WO 2016035032 6h and IL-8 supernatants were measured. Concentration-dependent inhibitory curves are shown in Figure 4 and in Table 3.

Table 3. Inhibition of IL-8 e in ed peripheral blood neutrophils from healthy (N=3) and COPD patients (N=3). Inhibitory concentration—dependent curves were generated by tion with Compound A (de A; 0.01nM-100uM) or Dexamethasone (DEX; 0.1nM- luM) in response to cigarette smoke extract (CSE 5%). Values are mean i SEM of 3 independent experiments run in triplicate. ICso values for half—maximum inhibition were calculated by nonlinear regression analysis. *p < 0.05 vs Healthy values.

Table 3 Stimulus HEALTHY COPD CSE 5% Maximal -log ICso N Maximal -log ICso N % Inhibition % Inhibition deA 97.4:53 6.53:0.22 851418.24 7.32:0.21 DEX 83.9::10.7 7.85::0.1719,84::11.46* 7.87:0.78 The addition of a ﬁxed concentration of dexamethasone 1 nM to increasing concentrations of Compound A of 0.1 nM, 1 nM, and 10 nM, showed increases in ting CSE-induced IL-8 release in phils from COPD patients (Figure 5).

Compound A concentration-dependently inhibited IL-8 secretion in neutrophils from healthy and COPD patients with a maximal percent inhibition of 97.4 :: .3% and 85.14 i 8.24% respectively. As a reference, the anti-inﬂammatory dexamethasone showed a favorable tory proﬁle on CSE-induced IL—8 release only in neutrophils from y patients with a maximal percent inhibition of 83.9 :: 10%. However in neutrophils from COPD patients, dexamethasone was not able to signiﬁcantly inhibit lL—8 release showing a corticosteroid insensitive proﬁle.

Example 7 Assay: Effect of nd A on basal RNA expression of osteroid resistance mediators and PI3K isoforms using eral blood neutrophils from healthy non- smokers and COPD smoker patients WO 2016035032 Measurement of basal RNA sion of corticosteroid ant mediators: Total RNA was isolated from peripheral human neutrophils from COPD patients in basal conditions and after experimental conditions. Cells were homogenized and RNA was extracted using e® Isolation Reagent (Roche, Indianapolis, USA). The reverse transcription was performed in 300ng of total RNA with TaqMan reverse transcription reagents kit (Applied Biosystems, Perkin-Elmer Corporation, CA, USA). 1.5 ul of result cDNA was ampliﬁed with speciﬁc predesigned primers (Applied Biosystems) for MIF (cat no 6988), MKP-l (cat no H500610256), PI3K-5 (cat n° Hs00192399), PI3Ky (cat n° Hs00277090) and GAPDH (cat no 4310884E) as endogenous control in a 7900HT Fast Real- Time PCR System (Applied Biosystem) using Universal Master Mix (Applied Biosystems).

Relative quantiﬁcation of these different transcripts was determined with the Z'AACt method and normalized to control . mRNA expression of the MIF, MKP-l, PI3K-5 and PI3Ky genes was measured in basal conditions and at the end of the experiments. ments were done in triplicate in at least three ts per experimental condition.

Results: The expression of MIF was not signiﬁcantly affected by CSE or Compound A exposure. In contrast, CSE decreased the expression of MKPl to approximately 04-fold of l. Compound A increased the expression of MKPl near to control levels which correlates well with the inhibitory effect of Compound A on E-S e. See Figure 6. While PI3K6 was not affected by CSE treatment, stration of Compound A caused a signiﬁcant reduction in CSE d PI3Ky expression (Figure 7).

Example 8 Assay: Effect of Compound A, Dexamethasone and combination thereof on basal expression of PI3K isoforms using peripheral blood neutrophils from y non-smokers and COPD smoker patients.

Measurement of Pi3K isoforms: To measure PI3K activity, neutrophils from COPD patients were isolated and incubated with Compound A at 10nM for 1h. Then cells were stimulated with CSE 5% for 30 min. After cell stimulation, neutrophils were centrifuged and total protein was extracted from neutrophils. Total n amount was measured using The Bio-Rad assay (Bio-Rad Laboratories Ltd., Herts, UK) to ensure equal amount. PI3K activity was measured using the PI3-Kinase Activity ELISA: Pico (cat. n° k- 1000s, Echelon Bioscience, Salt Lake City, USA) according to the cturer’s ol.

In brief, PI3-K reactions were run with the Class I PI3-K physiological substrate PI(4,5)P2 (PIP2). The enzyme reactions, PIP3 standards and controls were then mixed and incubated with PIP3 binding protein that is highly speciﬁc and sensitive to PIP3. This mixture was then erred to a PIP3-coated microplate for itive g. Afterwards, a peroxidase- linked ary detector and colorimetric detection was used to detect the amount of PIP3 produced by PI3 -K h comparing the enzyme reactions with a PIP3 standard curve.

Experiments were done in triplicate in at least three patients per experimental condition.

Results: In neutrophils from COPD patients, CSE 5% increased the PI3K activity measured as PIP3 production. The addition of Compound A at lOnM completely suppressed the PI3K ty (Figure 8)‘ Although the invention herein has been described with reference to particular embodiments, it is to be understood that these embodiments are merely rative of the principles and applications of the present invention. It is therefore to be understood that numerous modiﬁcations may be made to the illustrative embodiments and that other arrangements may be devised without departing from the spirit and scope of the present invention as described above. It is intended that the appended claims deﬁne the scope of the invention and that methods and ures within the scope of these claims and their equivalents be covered thereby.

All publications, patents and patent applications cited in this application are herein incorporated by reference to the same extent as if each individual publication, patent or patent application was speciﬁcally and individually indicated to be incorporated herein by reference.

Claims

WE CLAIM

1. Use of (i) a dual PI3K delta and gamma tor, and (ii) a corticosteroid in the manufacture of a medicament for treating an autoimmune, respiratory and/or inflammatory disease or condition, wherein the dual PI3K delta and gamma inhibitor and the corticosteroid are to be administered in a therapeutically ive amount, wherein the corticosteroid is selected from dexamethasone, fluticasone, asone propionate, mometasone furoate, and pharmaceutically acceptable salts thereof.

2. The use ing to claim 1, wherein the corticosteroid is selected from the group consisting of thasone, mometasone furoate, and fluticasone propionate and pharmaceutically acceptable salts thereof.

3. The use according to claims 1 or claim 2, wherein the corticosteroid is selected from the group consisting of dexamethasone, mometasone furoate, and fluticasone, and pharmaceutically acceptable salts thereof.

4. The use according to any one of claims 1-3, wherein the corticosteroid is selected from dexamethasone, fluticasone, and pharmaceutically able salts thereof.

5. The use according to any one of claims 1-4, wherein the therapeutically effective amount of (i) the dual PI3K delta and gamma inhibitor, and the therapeutically effective amount of (ii) the osteroid are to be administered simultaneously as a combined formulation.

6. The use according to any one of claims 1-5, n the therapeutically effective amount of (i) the dual PI3K delta and gamma tor, and the therapeutically effective amount of (ii) the corticosteroid are to be administered sequentially.

7. The use according to claim 6, wherein the therapeutically effective amount of the corticosteroid is to be administered before the therapeutically effective amount of the dual PI3K delta and gamma inhibitor.

8. The use according to any one of claims 1-7, wherein the therapeutically effective amount of the dual PI3K delta and gamma tor is to be administered twice daily to once every three weeks, and the therapeutically effective amount of the corticosteroid is to be administered twice daily to once every three weeks.

9. The use according to any one of claims 1-8, wherein the autoimmune, respiratory and/or inflammatory disease or condition is selected from the group consisting of asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, inflammatory bowel disease, glomerulonephritis, neuro matory diseases, multiple sclerosis, uveitis, psoriasis, arthritis, vasculitis, dermatitis, osteoarthritis, inflammatory muscle disease, allergic rhinitis, vaginitis, interstitial cystitis, scleroderma, osteoporosis, eczema, allogeneic or xenogeneic transplantation (organ, bone marrow, stem cells and other cells and tissues) graft ion, graft-versus-host disease, lupus erythematosus, inflammatory disease, type I diabetes, pulmonary fibrosis, dermatomyositis, Sjogren's syndrome, thyroiditis, myasthenia gravis, autoimmune hemolytic anemia, cystic fibrosis, idiopathic pulmonary fibrosis (IPF), chronic ing hepatitis, primary biliary cirrhosis, allergic ctivitis, atopic dermatitis, and combinations thereof.

10. The use according to any one of claims 1-9, wherein the autoimmune, respiratory and/or inflammatory disease or condition is selected from the group consisting of asthma, allergic rhinitis, non-allergic rhinitis, rheumatoid arthritis, c obstructive pulmonary disease, idiopathic ary fibrosis (IPF) and atopic dermatitis.

11. The use ing to any one of claims 1-10, wherein the dual PI3K delta and gamma tor and the corticosteroid are each to be administered in an amount ranging from about 0.01 mg to about 1000 mg.

12. The use of any one of claims 1-11, wherein the dual PI3K delta and gamma inhibitor and the corticosteroid are to be administered at a ratio of about 1:100 to about 100:1 by weight.

13. A pharmaceutical composition comprising (i) a dual PI3K delta and gamma inhibitor, (ii) a corticosteroid, and (iii) ally, a pharmaceutically acceptable carrier, t, diluent, or excipient, wherein the corticosteroid is selected from dexamethasone, fluticasone, fluticasone propionate, and mometasone furoate, and pharmaceutically acceptable salts thereof.

14. The pharmaceutical composition according to claim 13, wherein the corticosteroid is selected from the group consisting of thasone, mometasone furoate, and fluticasone propionate, and ceutically able salts thereof.

15. The pharmaceutical composition ing to claim 13 or claim 14, wherein the osteroid is selected from the group consisting of dexamethasone, mometasone furoate, and fluticasone, and pharmaceutically able salts thereof.

16. The pharmaceutical composition according to any one of claims 13-15, wherein the corticosteroid is selected from dexamethasone, fluticasone, and pharmaceutically acceptable salts thereof.

17. The pharmaceutical ition of any one of claims 13-16, wherein the composition comprises about 0.01 mg to about 1000 mg of the dual PI3K delta and gamma inhibitor and about 0.01 mg to about 1000 mg of the corticosteroid.

18. The pharmaceutical composition according to any one of claims 13-17, for use in a method of ng an autoimmune, atory and/or inflammatory disease or condition ed from the group consisting of asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, inflammatory bowel disease, glomerulonephritis, neuroinflammatory diseases, le sclerosis, uveitis, psoriasis, tis, vasculitis, dermatitis, osteoarthritis, inflammatory muscle disease, allergic rhinitis, vaginitis, interstitial cystitis, scleroderma, osteoporosis, eczema, neic or xenogeneic lantation (organ, bone marrow, stem cells and other cells and tissues) graft rejection, graft-versus-host disease, lupus erythematosus, inflammatory e, type I diabetes, pulmonary fibrosis, dermatomyositis, Sjogren's syndrome, thyroiditis (e.g., Hashimoto's and autoimmune thyroiditis), myasthenia gravis, autoimmune hemolytic anemia, cystic fibrosis, Idiopathic pulmonary fibrosis (IPF), chronic relapsing hepatitis, primary biliary cirrhosis, allergic conjunctivitis and atopic dermatitis, and combinations thereof.

19. The use of a pharmaceutical composition according to any one of claims 13-18 in the manufacture of a medicament for the treatment of autoimmune, respiratory and inflammatory diseases and conditions are selected from asthma, chronic obstructive ary disease, rheumatoid arthritis, inflammatory bowel e, glomerulonephritis, neuroinflammatory diseases, multiple sclerosis, uveitis, psoriasis, arthritis, vasculitis, dermatitis, osteoarthritis, inflammatory muscle disease, allergic rhinitis, vaginitis, interstitial cystitis, scleroderma, osteoporosis, eczema, allogeneic or neic transplantation (organ, bone marrow, stem cells and other cells and tissues) graft rejection, graft-versus-host disease, lupus erythematosus, inflammatory disease, type I diabetes, ary fibrosis, dermatomyositis, Sjogren's syndrome, thyroiditis (e.g., oto's and autoimmune thyroiditis), myasthenia gravis, autoimmune hemolytic anemia, cystic fibrosis, Idiopathic pulmonary fibrosis (IPF), chronic relapsing hepatitis, primary biliary cirrhosis, allergic conjunctivitis and atopic dermatitis, and combinations thereof.

20. A kit for treating an autoimmune, respiratory or inflammatory disease or condition, the kit comprising: (i) a dual PI3K delta and gamma inhibitor, and (ii) a corticosteroid, or a pharmaceutically acceptable salt thereof, in a single ceutical composition, (ii) ctions for treating the autoimmune, respiratory or inflammatory disease or condition with the dual PI3K delta and gamma inhibitor and corticosteroid and (iii) a container for placing the pharmaceutical composition or pharmaceutical compositions, wherein the corticosteroid is selected from dexamethasone, fluticasone, fluticasone propionate, and mometasone e, and pharmaceutically acceptable salts f.

21. The kit of claim 20, wherein the dual PI3K Delta and Gamma inhibitor and osteroid are for the treatment of an autoimmune, atory or inflammatory disease or condition selected from asthma, c obstructive pulmonary disease, rheumatoid arthritis, matory bowel disease, glomerulonephritis, neuroinflammatory diseases, multiple sclerosis, s, psoriasis, arthritis, vasculitis, dermatitis, osteoarthritis, inflammatory muscle disease, allergic rhinitis, vaginitis, interstitial cystitis, scleroderma, osteoporosis, eczema, allogeneic or xenogeneic transplantation (organ, bone marrow, stem cells and other cells and tissues) graft rejection, graft-versus-host e, lupus erythematosus, inflammatory disease, type I diabetes, pulmonary fibrosis, dermatomyositis, Sjogren's syndrome, thyroiditis (e.g., Hashimoto's and autoimmune thyroiditis), myasthenia gravis, autoimmune hemolytic anemia, cystic fibrosis, Idiopathic pulmonary is (IPF), chronic relapsing hepatitis, primary y cirrhosis, allergic conjunctivitis and atopic dermatitis.

22. The kit of claim 20 or claim 21, wherein the corticosteroid is selected from the group consisting of thasone, mometasone furoate, and fluticasone propionate, and pharmaceutically acceptable salts thereof.

23. The kit of any one of claims 20-22, wherein the corticosteroid is selected from the group consisting of dexamethasone, mometasone furoate, and fluticasone, and pharmaceutically acceptable salts thereof.

24. The kit of any one of claims 20-23, wherein the osteroid is selected from dexamethasone and fluticasone, and pharmaceutically acceptable salts thereof.