BFL-1 inhibitors

Abstract

The present invention provides compounds for use as inhibitors of BCL2A1 / BFL1 proteins and related methods, including methods of therapy which may include use in treating a disease or disorder characterized by inappropriate BCL2A1 / BFL1 expression and / or activity. In aspects, the disease or disorder is chorioamnionitis, an autoimmune disease or allergic disorder, or a cancer. Also provided are methods for attenuating a pathologic immune response.

Description

BFL-1 INHIBITORSFIELD OF THE INVENTION

[0001] The present invention relates to small molecule inhibitors of BFL-1 and related compositions and methods, including their use in methods for inhibiting BCL2A1 / BFL1 activity in a cell or tissue and methods of treating a disease or disorder characterized by inappropriate BCL2A1 / BFL1 expression and / or activity.BACKGROUND

[0002] Members of the BCL2 family of proteins are related through homology of their BCL2 homology (BH) domains. The proteins of this family may be divided into three main classes based on their activity in relation to cellular apoptosis: 1) anti-apoptotic or pro-survival factors; 2) pro-apoptotic activators or cytotoxic agents; and 3) pro-apoptotic sensitizers or effectors. Upon stress signaling, cells undergo apoptosis via mitochondrial outer membrane permeabilization (MOMP), which is driven by pro-apoptotic BCL2 family members. The pro- apoptotic BCL2 proteins are comprised of BH3-only proteins, such as NOXA, PUMA, and BIM that serve as sensitizers, and the multi-domain proteins, BAX and BAK that function as cytotoxic agents, promoting apoptosis. The BH3-only peptides either directly (through physical interaction) or indirectly (through interaction with anti-apoptotic Bcl-2 family members) activate BAX and BAK to promote mitochondrial membrane permeabilization and are upregulated during times of cell stress.

[0003] Pro-survival BCL2 proteins function through either prevention of BAX / BAK activity or sequestration of BH3-only activator proteins, resulting in inactive BAX / BAK. The prosurvival BCL2 family proteins sequester the pro-apoptotic members, driving a pro-survival phenotype and blocking MOMP. A number of family members have been described with varying targets and cell types in which they are present, including BCL2, BC1-XL, Bcl-w, MCL1, BCL-B and BCL2A1.

[0004] BFL1 is the human ortholog of murine BCL2A1. The two proteins share 72% amino acid sequence identity. BFL1 / BCL2A1 functions as a regulator of T-cell and neutrophil maturation and of the maintenance of CD4+T-cells. Knockout mice for all isoforms demonstrate decreased memory CD4+T cells, regulatory T-cells, and conventional dendritic cells in the spleen, but have an otherwise benign phenotype.BRIEF SUMMARY

[0005] Provided are compounds for use as inhibitors of Bcl2Al / Bfll . Accordingly, provided are methods for inhibiting BCL2A1 / BFL1 in a cell or tissue, the method comprising contacting the cell or tissue with a compound as described herein. In aspects, the compound is a compound of Formula I:whereinRi is oxo or -C(O)-Xi, wherein Xi is C1-3 alkoxy; R2 is halo or haloalkyl; R3 is halo or absent; and X and Y are each independently C, O, or S. In aspects, Ri is oxo or methyl formate; R2 is Cl or trifluoromethyl; R3 is F or absent; and / or X is C or O and Y is S or O. In aspects, the

[0006] In aspects, the compound is selected from the group consisting of

[0007] In aspects, the compound is selected from the group consisting of

[0008] In aspects, provided is a lipid nanoparticle comprising any one of the preceding compounds.

[0009] In one aspect, provided is a method for treating a disease or disorder characterized by inappropriate BCL2A1 / BFL1 expression and / or activity, where the method includes administering to a subject a pharmaceutical composition comprising any one of the preceding compounds.

[0010] In aspects, the method may also include where the disease or disorder is characterized by excessive inflammation.

[0011] In aspects, the method may also include where the disease or disorder is chorioamnionitis, an autoimmune disease or allergic disorder, or a cancer. In aspects, the cancer is adrenocortical carcinoma (ACC), acute myeloid leukemia (AML), bladder cancer, breast cancer, cervical cancer, cholangiocarcinoma, colorectal cancer, diffuse large B-cell (DLBC) lymphoma, glioblastoma multiforme (GBM), glioma, head and neck cancer, liver cancer, lung cancer, melanoma, mesothelioma, ovarian cancer, pheochromocytoma and paraganglioma (PCPG), pancreatic cancer, prostate cancer, sarcoma, testicular germ cell cancer, thymoma, thyroid cancer, uterine cancer, uveal melanoma, clear cell renal cell carcinoma (ccRCC), chromophobe renal cell carcinoma (chRCC), or papillary renal cell carcinoma (pRCC).

[0012] In aspects, the method may also include where the disease or disorder is anaphylaxis, asthma, atopic dermatitis, cystic fibrosis, hemophagocytic lymphohistiocytosis (HLH), irritablebowel syndrome (IBD), lupus erythematosus, multiple sclerosis, neutrophilic asthma, psoriasis, rheumatoid arthritis, or type 1 diabetes.

[0013] In aspects, the method may also include where the subject is human.

[0014] In aspects, the method may also include where the pharmaceutical compositions includes lipid nanoparticles encapsulating the compound. In aspects, the lipid nanoparticles comprise a targeting moiety.

[0015] In aspects, the targeting moiety is a mitochondrial targeting moiety. In aspects, the mitochondrial targeting moiety is a lipophilic cation with a delocalized positive charge such as the triphenylphosphonium ion (TPP), rhodamine, quaternary ammonium salts, pyridinium, cyanine and berberine derivatives, or dequalinium. In aspects, the mitochondrial targeting moiety is a mitochondrial penetrating peptide (MPP), also referred to as Szeto-Schiller peptides.

[0016] In aspects, the targeting moiety is a polypeptide or antibody, or an antigen binding fragment thereof, that binds to a neutrophil-specific cell surface glycoprotein. In aspects, the neutrophil-specific cell surface glycoprotein is a cluster of differentiation (“CD”) protein including CD177 and CD66b, signal regulatory protein alpha (“SIRPa”), or sialic acid-binding Ig-like lectin 9 (SIGLEC9). In aspects, the targeting moiety is an antibody or antigen binding fragment thereof that targets proteinase 3 (“PRTN3”).

[0017] Other technical features may be readily apparent to one skilled in the art from the following figures, descriptions, and claims.BRIEF DESCRIPTION OF THE FIGURES

[0018] FIG. 1A shows single-dose inhibition data (normalized fluorescence polarization, FP) for 48 compounds, where 100 represents fully bound NOXA BH3 peptide and 0 represents completely displaced (0% bound) NOXA BH3 peptide.

[0019] FIG. IB shows inhibition data (normalized FP) for the remaining compounds.

[0020] FIG. 2 shows FP dose response curves for Compound 1, Compound 2, Compound 3, Compound 4, Compound 6, and Compound 7.DETAILED DESCRIPTION

[0021] The present disclosure provides compounds and related compositions and methods for inhibiting BCL2A1 / BFL1 protein activity. The murine (BCL2A1) and human (BFL1) proteinsare highly conserved, particularly in their BH3 binding domain, or “BH3 groove”. Accordingly, in the context of the present disclosure, inhibitors of the murine BCL2A1 are reasonably expected to inhibit the human BFL1 to a similar degree.Compounds

[0022] Definitions

[0023] “Alkyl” refers to a straight or branched, saturated, aliphatic radical having the number of carbon atoms indicated. Alkyl can include any number of carbons, such as C1-2, C1-3, C1-4, C 1-5, C 1-6, C1-7, C1-8, C 1-9, Ci-10, C2-3, C2-4, C2-5, C2-6, C3-4, C3-5, C3-6, C4-5, C4-6 and C5-6. For example, C1-6 alkyl includes, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, hexyl, etc. Alkyl can also refer to alkyl groups having up to 20 carbons atoms, such as, but not limited to heptyl, octyl, nonyl, decyl, etc. Alkyl groups can be substituted or unsubstituted.

[0024] “Alkylene” refers to a straight or branched, saturated, aliphatic radical having the number of carbon atoms indicated, and linking at least two other groups, i.e., a divalent hydrocarbon radical. The two moi eties linked to the alkylene can be linked to the same atom or different atoms of the alkylene group. For instance, a straight chain alkylene can be the bivalent radical of — (CH2)n — , where n is 1, 2, 3, 4, 5 or 6. Representative alkylene groups include, but are not limited to, methylene, ethylene, propylene, isopropylene, butylene, isobutylene, secbutylene, pentylene and hexylene. Alkylene groups can be substituted or unsubstituted.

[0025] “Alkoxy” refers to an alkyl group having an oxygen atom that connects the alkyl group to the point of attachment: alkyl-0 — . As for alkyl group, alkoxy groups can have any suitable number of carbon atoms, such as C1-6. Alkoxy groups include, for example, methoxy, ethoxy, propoxy, iso-propoxy, butoxy, 2-butoxy, iso-butoxy, sec-butoxy, tert-butoxy, pentoxy, hex oxy, etc. The alkoxy groups can be further substituted with a variety of substituents described within. Alkoxy groups can be substituted or unsubstituted.

[0026] “Hydroxyalkyl” refers to an alkyl group, as defined above, where at least one of the hydrogen atoms is replaced with one or more hydroxy group. As for the alkyl group, hydroxyalkyl groups can have any suitable number of carbon atoms, such as C. sub.1-8. Exemplary hydroxyalkyl groups include, but are not limited to, hydroxy-methyl, hydroxyethyl (where the hydroxy is in the 1- or 2-position), hydroxypropyl (where the hydroxy is in the 1-, 2- or 3-position), 2,3-dihydroxypropyl, hydroxybutyl (where the hydroxy is in the 1-, 2-, 3- or4-position), hydroxypentyl (where the hydroxy is in the 1-, 2-, 3-, 4- or 5-position), hydroxyhexyl (where the hydroxy is in the 1-, 2-, 3-, 4-, 5- or 6-position), 1,2-dihydroxy ethyl, and the like.

[0027] “Haloalkyl” refers to alkyl, as defined above, where one, some, or all of the hydrogen atoms are replaced with halogen atoms. As for alkyl group, haloalkyl groups can have any suitable number of carbon atoms, such as C. sub.1-8. For example, haloalkyl includes trifluoromethyl, flouromethyl, etc.

[0028] “Halogen” or “halo” refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

[0029] “ Oxo” refers to an oxygen atom connected to the point of attachment by a double bond (=O).

[0030] Provided are compounds for use as inhibitors of BCL2A1 / BFL1 proteins. In aspects, the compound is a compound of Formula I:whereinRi is oxo or -C(0)-Xi, wherein Xi is C1-3 alkoxy; R2 is halo or haloalkyl; R3 is halo or absent; and X and Y are each independently C, O, or S. In aspects, Ri is oxo or methyl formate; R2 is Cl or trifluoromethyl; R3 is F or absent; and / or X is C or O and Y is S or O. In aspects, the

[0031] In aspects, the compound is selected from the group consisting of

[0032] In aspects, provided is a compound for use as a BCL2A1 / BFL1 inhibitor selected from the group consisting ofBiological Activity

[0033] A panel of 96 compounds (available, e.g., from Enamine Ltd and other commercial sources) was screened for biological activity using a fluorescence polarization (FP) assay. Assays were carried out using fluorescently labeled NOXA peptide (FITC-labeled mouse NOXA, mNoxa; Peptide 2.0) which binds within the BH3-binding groove. The FP assay measures the tumbling rate of the labeled NOXA peptide as a proxy for inhibition of BH3 binding using a competition-based approach. When NOXA is bound to BCL2A1, its rate of tumbling is low (and FP is high), whereas an effective inhibitor compound will displace NOXA, causing a fast rate of tumbling (and low FP value).

[0034] Briefly, FP assays were performed in two steps: single-point high-concentration compounds and dose response of fluorescence polarization hits. BCL2A1 was added at 3 pM to 200 pM or 400 pM of each compound in 20 mM Tris pH 7, 500 mM NaCl, 0.005 % Tween-20 buffer. After addition of 375 nM labeled mNOXA, 96-well plates were incubated overnight at 20°C in the dark to achieve equilibrium before fluorescence polarization was measured with a Biotek Synergy H2. Autofluorescent compounds and fluorescent quenching compounds were corrected via ratiometric correction as described by Shapiro et al., 2009. Any compounds that showed a significant shift in polarization, along with those identified in the thermal shift assays had a dose response measured via FP.

[0035] Dose response curves were measured by adding 3 pM BCL2A1 to a serial two-fold dilution series of each compound ranging from 400 pM to 781 nM in 20 mM Tris pH 7, 500 mM NaCl, 0.005% Tween-20 buffer. Some compounds were further tested to assess the accuracy of these two-fold dilutions with serial 1.33-fold dilutions. BCL2A1, compounds, buffer and lastly, 375 nM FITC-labeled mNOXA were added to each well and incubated in the dark at 20°C overnight to achieve equilibrium, followed by measurement of polarization. All dose responses were performed in triplicate.

[0036] FIG. 1 A and FIG. IB show biological activity (normalized FP) for the 96 compounds tested.

[0037] FIG. 2 shows representative FP dose response curves for Compound 1, Compound 2, Compound 3, Compound 4, Compound 6, and Compound 7.

[0038] IC50 values are given in Table 1.Table 1 : IC 50 values for competitive inhibition in the FP assayIC50 KiCompound IC50 (pM) Ki (pM) Compound(pM) (pM)1 129 2.53 19 1002 19.652 182 3.57 20 424 8.313 157 3.08 21 674 13.224 321 6.29 22 1181 23.165 1257 24.65 23 1331 26.106 362 7.10 24 534 10.477 418 8.20 25 1173 23.008 3218 63.10 26 414 8.129 810 15.88 27 639 12.5310 877 17.20 28 1391 27.2711 756 14.82 29 1445 28.3312 6505 127.55 30 1326 26.0013 381 7.47 31 3749 73.5114 363 7.12 32 823 16.1415 517 10.14 33 879 17.2416 553 10.84 34 1691 33.1617 683 13.39 35 1693 33.2018 656 12.86 36 10910 213.9Methods

[0039] The inventors previously discovered a significant upregulation of BCL2A1 / BFL1 in activated immune cells, including T cells, neutrophils, and dendritic cells. Importantly, BCL2A1 / BFL1 protein is specifically upregulated in the activated immune cells but not resting immune cells. In addition, knockout mice lacking all three genes of the Bcl2Al locus showed minimal effects on tissue homeostasis. Schenk RL Cell Death & Diff. 2017. Together, these data indicate that anti-Al therapy is likely to be safe and effective.

[0040] Provided are compounds for use in methods for treating a disease or disorder characterized by inappropriate BCL2A1 / BFL1 expression and / or activity. In aspects, the disease or disorder is characterized by excessive inflammation, particularly that may be mediated by neutrophils, T cells, dendritic cells, and combinations thereof. In aspects, the disease or disorder is chorioamnionitis, an autoimmune disease or allergic disorder characterized by neutrophil-mediated inflammation, or a cancer characterized by theoverexpression of BFL1. Also provided are methods for attenuating a pathologic immune response in a subject in need of such treatment, the method comprising administering to the subject a compound as described herein.

[0041] In aspects, provided are methods of therapy comprising administering to a subject in need of such therapy a compound as described herein, or a pharmaceutical composition comprising a compound as described herein. In aspects, the method of therapy is one for treating a disease or disorder characterized by inappropriate BFL1 activity. In aspects, the disease or disorder is characterized by excessive inflammation, particularly neutrophil- mediated inflammation. In aspects, the disease or disorder is chorioamnionitis, an autoimmune disease or allergic disorder characterized by neutrophil-mediated inflammation, or a cancer characterized by the overexpression of BFL1. In aspects, the method of therapy is one for attenuating a pathologic immune response in the subject. In aspects, while attenuating a pathologic immune response, the method results in only a modest global immune suppression.

[0042] In accordance with any of the preceding uses or methods of therapy, in aspects the disease or disorder is anaphylaxis, asthma, atopic dermatitis, cystic fibrosis, hemophagocytic lymphohistiocytosis (HLH), irritable bowel syndrome (IBD), including more specifically one or both of Crohn’s disease and ulcerative colitis, lupus erythematosus, multiple sclerosis, neutrophilic asthma, psoriasis, rheumatoid arthritis, or type 1 diabetes. In aspects, the disease or disorder is lymphohistiocytosis (HLH), multiple sclerosis, neutrophilic asthma, or type 1 diabetes.

[0043] In accordance with any of the preceding uses or methods of therapy, in aspects the disease or disorder is a cancer selected from adrenocortical carcinoma (ACC), acute myeloid leukemia (AML), bladder cancer, breast cancer, cervical cancer, cholangiocarcinoma, colorectal cancer, diffuse large B-cell (DLBC) lymphoma, glioblastoma multiforme (GBM), glioma, head and neck cancer, liver cancer, lung cancer, melanoma, mesothelioma, ovarian cancer, pheochromocytoma and paraganglioma (PCPG), pancreatic cancer, prostate cancer, sarcoma, testicular germ cell cancer, thymoma, thyroid cancer, uterine cancer, uveal melanoma, clear cell renal cell carcinoma (ccRCC), chromophobe renal cell carcinoma (chRCC), and papillary renal cell carcinoma (pRCC).

[0044] In accordance with any of the preceding uses or methods of therapy, in aspects the disease or disorder is a cancer selected from acute myeloid leukemia (AML), breast cancer, diffuse large B-cell lymphoma (DLBCL), head and neck cancer, lung cancer, melanoma,mesothelioma, ovarian cancer, pancreatic cancer, testicular germ cell cancer, thymoma, uterine cancer, clear cell renal cell carcinoma (ccRCC), chromophobe renal cell carcinoma (chRCC), and papillary renal cell carcinoma (pRCC).

[0045] In the context of the methods described here, the term “treating” may refer to the amelioration or stabilization of one or more symptoms associated with the disease, disorder or condition being treated. The term “treating” may also encompass the management of disease, disorder or condition, referring to the beneficial effects that a subject derives from a therapy but which does not result in a cure of the underlying disease, disorder, or condition. In the context of the present disclosure, the term “prevention” refers to preventing the recurrence, development, progression or onset of one or more symptoms of the disease, disorder, or condition.

[0046] In embodiments where a therapeutically effective amount of a compound or composition is administered to a subject, the therapeutically effective amount is the amount sufficient to achieve a desired therapeutic outcome, for example the amelioration or stabilization of one or more symptoms of the disease, disorder or condition being treated, or in the context of prevention, the amount sufficient to achieve prevention of the recurrence, development, progression or onset of one or more symptoms of the disease, disorder, or condition.

[0047] In aspects, a therapeutically effective amount is the amount of a compound required to achieve at least an equivalent therapeutic effect compared to a standard therapy. An example of a standard therapy is an FDA-approved drug indicated for treating the same disease, disorder or condition.

[0048] In the context of any of the methods described here, the subject is preferably a human but may be a non-human vertebrate. In aspects, the non-human vertebrate may be, for example, a dog, cat, a rodent (e.g., a mouse, a rat, a rabbit), a horse, a cow, a sheep, a goat, a chicken, a duck, or any other non-human vertebrate.

[0049] Accordingly, in aspects the subject is a human subject. In aspects, the human subject is selected from an adult human, a pediatric human, or a geriatric human, as those terms are understood by the medical practitioner, for example as defined by the U.S. Food and Drug Administration.

[0050] In aspects, the human subject is one in need of therapy for a disease or disorder characterized by inappropriate BFL1 activity. In aspects, the disease or disorder ischaracterized by excessive inflammation, particularly neutrophil-mediated inflammation. In aspects, the disease or disorder is chorioamnionitis, an autoimmune disease or allergic disorder characterized by neutrophil-mediated inflammation, or a cancer characterized by the overexpression of BFL1. In aspects, the human subject is one in need of therapy for attenuating a pathologic immune response.Compositions

[0051] Also provided are pharmaceutical compositions for use in treating a disease or disorder characterized by inappropriate BCL2A1 / BFL1 expression and / or activity comprising a compound as described herein, and one or more pharmaceutically acceptable excipients or carriers.

[0052] In aspects, compositions comprising a compound as described herein are adapted for targeted in vivo delivery. In aspects, provided are compositions targeted for delivery to activated tissue neutrophils. Targeted delivery systems that may be used include nanoparticles. In aspects, the nanoparticles may be liposomes, polymers, dendrimers, or magnetic nanoparticles. Nanoparticulate delivery systems suitable for targeting a BCL2A1 / BFL1 inhibitor to activated tissue neutrophils include liposome-based nanoparticles such as those described in Gautam et al J. Drug Delivery Sci. Tech. 2017 260-268 and Peer et al. Science 2008 319(5863):627-30. For example, the liposome-based nanoparticles may comprise nanoparticular sized (50-500 nm diameter) liposomes formed from neutral phospholipids comprising a glycosaminoglycan such as hyaluronan to which a targeting moiety is attached.

[0053] In aspects, the targeting moiety may be a mitochondrial targeting moiety. In aspects, the mitochondrial targeting moiety is a lipophilic cation with a delocalized positive charge such as the triphenylphosphonium ion (TPP), rhodamine, quaternary ammonium salts, pyridinium, cyanine and berberine derivatives, or dequalinium. In aspects, the mitochondrial targeting moiety is a mitochondrial penetrating peptide (MPP), also referred to as Szeto-Schiller peptides, containing aromatic and positively charged residues. General features of MPPs include a highly hydrophobic residue such as Fx (cyclohexylalanine) and an alternatively placed positively charged moiety, such as arginine (R) or lysine (K).

[0054] In aspects, provided are lipid nanoparticles, also referred to as “liposomes” loaded with a BCL2A1 / BFL1 inhibitor as described herein. In aspects, the compound is encapsulated in a liposome-based nanoparticle comprising a targeting moiety. In aspects, the targetingmoiety is a polypeptide, an antibody, or an antigen-binding fragment of a polypeptide or antibody. In aspects, the polypeptide, antibody, or binding fragment thereof binds to a neutrophil-specific cell surface glycoprotein. In aspects, the neutrophil-specific cell surface glycoprotein is a cluster of differentiation (“CD”) protein including CD 177 and CD66b, signal regulatory protein alpha (“SIRPa”), or sialic acid-binding Ig-like lectin 9 (SIGLEC9). Thus, in aspects, the cell surface glycoprotein is CD 177, CD66b, SIRPa, or SIGLEC9. In aspects, the targeting moiety is an antibody or antigen binding fragment thereof that targets proteinase 3 (“PRTN3”).

[0055] The term “pharmaceutically acceptable” refers to those compounds, materials, compositions, carriers, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. Excipients for preparing a pharmaceutical composition are generally those that are known to be safe and non-toxic when administered to a human or animal body. Examples of pharmaceutically acceptable excipients include, without limitation, sterile liquids, water, buffered saline, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol and the like), oils, detergents, suspending agents, carbohydrates (e.g., glucose, lactose, sucrose or dextran), antioxidants (e.g, ascorbic acid or glutathione), chelating agents, low molecular weight proteins, and suitable mixtures of any of the foregoing. The particular excipients utilized in a composition will depend upon various factors, including chemical stability and solubility of the compound being formulated and the intended route of administration.

[0056] A pharmaceutical composition can be provided in bulk or unit dosage form. It is especially advantageous to formulate pharmaceutical compositions in unit dosage form for ease of administration and uniformity of dosage. The term “unit dosage form” refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of an active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. A unit dosage form can be an ampoule, a vial, a suppository, a dragee, a tablet, a capsule, an IV bag, or a single pump on an aerosol inhaler.

[0057] In therapeutic applications, dose may vary depending on the chemical and physical properties of the active compound as well as clinical characteristics of the subject, includinge.g., age, weight, and co-morbidities. Generally, the dose should be a therapeutically effective amount. An effective amount of a pharmaceutical composition is that which provides an objectively identifiable improvement as noted by the clinician or other qualified observer. For example, alleviating a symptom of a disorder, disease or condition.

[0058] A pharmaceutical compositions may take any suitable form (e.g. liquids, aerosols, solutions, inhalants, mists, sprays; or solids, powders, ointments, pastes, creams, lotions, gels, patches and the like) for administration by any desired route (e.g. pulmonary, inhalation, intranasal, oral, buccal, sublingual, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, intrapleural, intrathecal, transdermal, transmucosal, rectal, and the like). In embodiments, the pharmaceutical composition is in the form of an orally acceptable dosage form including, but not limited to, capsules, tablets, buccal forms, troches, lozenges, and oral liquids in the form of emulsions, aqueous suspensions, dispersions or solutions. Capsules may contain excipients such as inert fillers and / or diluents including starches (e.g., corn, potato or tapioca starch), sugars, artificial sweetening agents, powdered celluloses, such as crystalline and microcrystalline celluloses, flours, gelatins, gums, etc. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, can also be added.

[0059] In embodiments, the pharmaceutical composition is in the form of a tablet. The tablet can comprise a unit dose of a compound described here together with an inert diluent or carrier such as a sugar or sugar alcohol, for example lactose, sucrose, sorbitol or mannitol. The tablet can further comprise a non-sugar derived diluent such as sodium carbonate, calcium phosphate, calcium carbonate, or a cellulose or derivative thereof such as methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, and starches such as corn starch. The tablet can further comprise binding and granulating agents such as polyvinylpyrrolidone, disintegrants (e.g. swellable crosslinked polymers such as crosslinked carboxymethylcellulose), lubricating agents (e.g. stearates), preservatives (e.g. parabens), antioxidants (e.g. butylated hydroxytoluene), buffering agents (e.g. phosphate or citrate buffers), and effervescent agents such as citrate / bicarbonate mixtures. The tablet may be a coated tablet. The coating can be a protective film coating (e.g. a wax or varnish) or a coating designed to control the release of the active compound, for example a delayed release (release of the active after a predetermined lag time following ingestion) or release at a particular location in the gastrointestinal tract. The lattercan be achieved, for example, using enteric film coatings such as those sold under the brand name Eudragit®.

[0060] Tablet formulations may be made by conventional compression, wet granulation or dry granulation methods and utilize pharmaceutically acceptable diluents, binding agents, lubricants, disintegrants, surface modifying agents (including surfactants), suspending or stabilizing agents, including, but not limited to, magnesium stearate, stearic acid, talc, sodium lauryl sulfate, microcrystalline cellulose, carboxymethylcellulose calcium, polyvinylpyrrolidone, gelatin, alginic acid, acacia gum, xanthan gum, sodium citrate, complex silicates, calcium carbonate, glycine, dextrin, sucrose, sorbitol, dicalcium phosphate, calcium sulfate, lactose, kaolin, mannitol, sodium chloride, talc, dry starches and powdered sugar. Preferred surface modifying agents include nonionic and anionic surface modifying agents.Representative examples of surface modifying agents include, but are not limited to, poloxamer 188, benzalkonium chloride, calcium stearate, cetostearyl alcohol, cetomacrogol emulsifying wax, sorbitan esters, colloidal silicon dioxide, phosphates, sodium dodecyl sulfate, magnesium aluminum silicate, and triethanolamine.

[0061] In embodiments, the pharmaceutical composition is in the form of a hard or soft gelatin capsule. In accordance with this formulation, the compound of the present invention may be in a solid, semi-solid, or liquid form.

[0062] In embodiments, the pharmaceutical composition is in the form of a sterile aqueous solution or dispersion suitable for parenteral administration. The term parenteral as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intra- articular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.

[0063] In embodiments, the pharmaceutical composition is in the form of a sterile aqueous solution or dispersion suitable for administration by either direct injection or by addition to sterile infusion fluids for intravenous infusion, and comprises a solvent or dispersion medium containing, water, ethanol, a polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), suitable mixtures thereof, or one or more vegetable oils. Solutions or suspensions can be prepared in water with the aid of co-solvent or a surfactant. Examples of suitable surfactants include polyethylene glycol (PEG)-fatty acids and PEG-fatty acid mono and diesters, PEG glycerol esters, alcohol-oil transesterification products, polyglyceryl fatty acids, propylene glycol fatty acid esters, sterol and sterol derivatives, polyethylene glycol sorbitanfatty acid esters, polyethylene glycol alkyl ethers, sugar and its derivatives, polyethylene glycol alkyl phenols, polyoxyethylene-polyoxypropylene (POE-POP) block copolymers, sorbitan fatty acid esters, ionic surfactants, fat-soluble vitamins and their salts, water-soluble vitamins and their amphiphilic derivatives, amino acids and their salts, and organic acids and their esters and anhydrides. Dispersions can also be prepared, for example, in glycerol, liquid polyethylene glycols and mixtures of the same in oils.

[0064] In embodiments, a compound or composition described here may be administered as monotherapy or adjunctive therapy. In embodiments, a compound or composition described here may be administered alone or in combination with one or more additional therapeutic agents (z.e., additional APIs) or therapies, for example as part of a therapeutic regimen that includes, e.g., aspects of diet and exercise). In embodiments, the methods described here include administration of compound of Formula I, or a derivative thereof, as the primary therapy. In other embodiments, the administration of compound of Formula I, or a derivative thereof, is an adjuvant therapy. In either case, the methods of the invention contemplate the administration of compound of Formula I, or a derivative thereof, in combination with one or more additional therapeutic agents and / or therapies for the treatment or prevention of a disease, disorder, or condition as described here. The terms “therapy” and “therapies” refer to any method, protocol and / or agent that can be used in the prevention, treatment, management or amelioration of a disease, disorder, or condition, one or more symptoms thereof.

[0065] The present disclosure also provides packaging and kits comprising pharmaceutical compositions for use in the methods described here. The kit can comprise one or more containers selected from the group consisting of a bottle, a vial, an ampoule, a blister pack, and a syringe. The kit can further include one or more of instructions for use, one or more syringes, one or more applicators, or a sterile solution suitable for reconstituting a compound or composition described here

[0066] Unless defined otherwise, technical terms used herein are to be understood as commonly understood by one of ordinary skill in the art to which the disclosure belongs.

[0067] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art. See, e.g., Singleton et al., Dictionary of Microbiology and Molecular Biology, 2nd ed., J. Wiley & Sons (New York, NY 1994); Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989). Any methods, devices and materials similar orequivalent to those described herein can be used in the practice of this disclosure. The following definitions are provided to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure.

[0068] The phrases “at least one”, “one or more”, and “and / or”, as used herein, are open- ended expressions that are both conjunctive and disjunctive in operation. The terms “a”, “an”, “the”, “first”, “second”, etc., do not preclude a plurality. For example, the term “a” or “an” entity, as used herein, refers to one or more of that entity. As such, the terms “a” (or “an”), “one or more” and “at least one” can be used interchangeably herein.

[0069] The term “about” when used before a numerical designation, e.g, temperature, time, amount, concentration, and such other, including a range, indicates approximations which may vary by ( + ) or ( - ) 10%, 5%, 1%, or any subrange or subvalue there between. Preferably, the term “about” means that the value may vary by + / - 10%.

[0070] The term “comprises / comprising” does not exclude the presence of other elements, components, features, regions, integers, steps, operations, etc. Additionally, although individual features may be included in different claims, these may possibly advantageously be combined, and the inclusion in different claims does not imply that a combination of features is not feasible and / or advantageous. By contrast, the transitional phrase “consisting of’ excludes any element, step, or ingredient not specified in the claim. The transitional phrase “consisting essentially of’ limits the scope of a claim to the specified materials or steps “and those that do not materially affect the basic and novel characteristic(s)” of the claimed invention.

[0071] While the invention herein disclosed has been described by means of specific embodiments and applications thereof, numerous modifications and variations could be made thereto by those skilled in the art without departing from the scope of the invention set forth in the claims.

[0072] It will be appreciated that the present invention is set forth in various levels of detail in this application. In certain instances, details that are not necessary for one of ordinary skill in the art to understand the invention, or that render other details difficult to perceive may have been omitted. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting beyond the scope of the appended claims.

Claims

CLAIMSWhat is claimed is:

1. A method for inhibiting BCL2A1 / BFL1 in a cell or tissue, in vivo, in vitro, or ex vivo, the method comprising contacting the cell or tissue with a compound of Formula IRi is oxo or -C(O)-Xi, wherein Xi is C1-3 alkoxy;R2 is halo or haloalkyl;R3 is halo or absent; andX and Y are each independently C, O, or S.

2. The method of claim 1, wherein Ri is oxo or methyl formate.

3. The method of claim 1 or 2, wherein R2 is Cl or trifluoromethyl.

4. The method of any one of claims 1 to 3, wherein R3 is F or absent.

5. The method of any one of claims 1 to 4, wherein X is C or O and Y is S or O.

6. The method of any one of claims 1 to 5, wherein the compound is7. A method for inhibiting BCL2A1 / BFL1 in a cell or tissue, in vivo, in vitro, or ex vivo, the method comprising contacting the cell or tissue with a compound selected from the group consisting of8. A method for inhibiting BCL2A1 / BFL1 in a cell or tissue, in vivo, in vitro, or ex vivo, the method comprising contacting the cell or tissue with a compound selected from the group consisting of9. The method of any one of claims 1 to 8, wherein the compound is encapsulated in a lipid nanoparticle.

10. The method of claim 9, wherein the lipid nanoparticle comprises a targeting moiety.

11. The method of claim 10, wherein the targeting moiety is a mitochondrial penetrating peptide (MPP), triphenylphosphonium ion (TPP), rhodamine, a quaternary ammonium salt, pyridinium, cyanine or a derivative thereof, berberine or a derivative thereof, or dequalinium.

12. The method of claim 10, wherein the targeting moiety is a mitochondrial penetrating peptide (MPP) or triphenylphosphonium ion (TPP).

13. The method of claim 10, wherein the targeting moiety is a polypeptide or antibody, or an antigen binding fragment thereof, that binds to a neutrophil-specific cell surface glycoprotein.

14. The method of claim 13, wherein the neutrophil-specific cell surface glycoprotein is a cluster of differentiation (“CD”) protein including CD 177 and CD66b, signal regulatory protein alpha (“SIRPa”), or sialic acid-binding Ig-like lectin 9 (SIGLEC9).

15. The method of claim 10, wherein the targeting moiety is an antibody or antigen binding fragment thereof that targets proteinase 3 (“PRTN3”).

16. A method for treating a disease or disorder characterized by inappropriate BCL2A1 / BFL1 expression and / or activity, the method comprising administering to a subject a pharmaceutical composition comprising the compound as described in any one of claims 1 to 15.

17. The method of claim 16, wherein the disease or disorder is characterized by excessive inflammation.

18. The method of claim 16, wherein the disease or disorder is chorioamnionitis, an autoimmune disease or allergic disorder, or a cancer.

19. The method of claim 18, wherein the cancer is adrenocortical carcinoma (ACC), acute myeloid leukemia (AML), bladder cancer, breast cancer, cervical cancer, cholangiocarcinoma, colorectal cancer, diffuse large B-cell (DLBC) lymphoma, glioblastoma multiforme (GBM), glioma, head and neck cancer, liver cancer, lung cancer, melanoma, mesothelioma, ovarian cancer, pheochromocytoma and paraganglioma (PCPG), pancreatic cancer, prostate cancer, sarcoma, testicular germ cell cancer, thymoma, thyroid cancer, uterine cancer, uveal melanoma, clear cell renal cell carcinoma (ccRCC), chromophobe renal cell carcinoma (chRCC), or papillary renal cell carcinoma (pRCC).

20. The method of claim 16, wherein the disease or disorder is anaphylaxis, asthma, atopic dermatitis, cystic fibrosis, hemophagocytic lymphohistiocytosis (HLH), irritable bowel syndrome (IBD), lupus erythematosus, multiple sclerosis, neutrophilic asthma, psoriasis, rheumatoid arthritis, or type 1 diabetes.

21. The method of any one of claims 16 to 20, wherein the subject is human.

Images