WO2016131100A1 - Méthodes de traitement de maladies infectieuses - Google Patents

Méthodes de traitement de maladies infectieuses Download PDF

Info

Publication number
WO2016131100A1
WO2016131100A1 PCT/AU2016/050105 AU2016050105W WO2016131100A1 WO 2016131100 A1 WO2016131100 A1 WO 2016131100A1 AU 2016050105 W AU2016050105 W AU 2016050105W WO 2016131100 A1 WO2016131100 A1 WO 2016131100A1
Authority
WO
WIPO (PCT)
Prior art keywords
infection
bcl2 family
bcl
formula
agent
Prior art date
Application number
PCT/AU2016/050105
Other languages
English (en)
Inventor
James Vince
Thomas NADERER
Mary Helen SPEIR
Marc Pellegrini
Samar OJAIMI
Kathryn Elizabeth Lawlor
Original Assignee
The Walter And Eliza Hall Institute Of Medical Research
Monash University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from AU2015900554A external-priority patent/AU2015900554A0/en
Application filed by The Walter And Eliza Hall Institute Of Medical Research, Monash University filed Critical The Walter And Eliza Hall Institute Of Medical Research
Publication of WO2016131100A1 publication Critical patent/WO2016131100A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
    • A61K31/381—Heterocyclic compounds having sulfur as a ring hetero atom having five-membered rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/426—1,3-Thiazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/428—Thiazoles condensed with carbocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/472—Non-condensed isoquinolines, e.g. papaverine
    • A61K31/4725—Non-condensed isoquinolines, e.g. papaverine containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A61P31/06—Antibacterial agents for tuberculosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/10—Antimycotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • A61P31/22—Antivirals for DNA viruses for herpes viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the specification relates generally to the field of therapeutic agents. More particularly, the specification relates to methods and related compositions for preventing or treating infectious diseases.
  • Infectious disease persists as a worldwide health challenge.
  • pathogens e.g., antibiotic-resistant pathogenic bacteria
  • antimicrobial resistance largely driven by overuse of antibiotics, is now considered a global health threat.
  • BCL2 family inhibitors can be used to treat infections in a subject. Accordingly, in one aspect described herein is a method for treating an infection, comprising administering to a subject in need thereof a therapeutically effective amount of a BCL2 family inhibitor.
  • the infection to be treated is a bacterial infection.
  • the bacterial infection is an infection by bacteria of one or more species of Legionella, Mycobacterium, Salmonella, Staphylococcus, pathogenic E. coli, or Pseudomonas.
  • the bacterial infection includes infection by antibiotic-resistant bacteria.
  • treatment of a subject suffering from a bacterial infection includes, in addition to treatment with a BCL2 family inhibitor, treatment with a bactericidal or bacteriostatic agent.
  • the bactericidal or bacteriostatic agent is administered contemporaneously with the BCL2 family inhibitor.
  • the BCL2 family inhibitor is administered before the bactericidal or bacteriostatic agent.
  • the BCL2 family inhibitor is administered after the bactericidal or bacteriostatic agent
  • the infection to be treated is a viral infection, in particular a viral infection other than a viral infection by an influenza virus.
  • the viral infection includes infection by one or more of, HSV, HTLV-1, HTLV-2, HIV, a respiratory virus, rhinovirus, or a coronavirus, or Ebola.
  • the infection to be treated is a fungal infection.
  • the fungal infection is an infection by one or more of a yeast, Aspergillus, Pneumocystitis, and Cryptococcus.
  • the infection to be treated is a protozoal infection.
  • the protozoal infection is an infection by one or more of Trypanosomoa cruzi, Toxoplasma gondii, Cryptosporidium, or Encephalitozoon.
  • the subject is suffering from tuberculosis, pneumonia, Legionnaires' disease, or an acute respiratory infection.
  • subject to be treated is suffering from an infection that includes infection of macrophages.
  • the infected macrophages include alveolar macrophages.
  • the infection is an acute or subacute infection.
  • the subject to be treated for the infection was not diagnosed as suffering from a cancer.
  • administration of the BCL2 family inhibitor occurs within a period from about 1 hours to about 72 hours, about 6 hours to about 48 hours , or about 6 hours to about 24 hours, following a diagnosis of the infection. In other embodiments, the administration occurs no later than about 72 hours or about 48 hours following a diagnosis of the infection.
  • the subject to be treated for the infection is human. In other embodiments the subject to be treated is non- human. In some embodiments the non- human subject is a non- human primate, a rat, a mouse, a pig, a cow, a sheep, a chicken, or a duck.
  • the inhibitor at least targets one or more or all of BCL-XL, BCL-2, MCL-1 or Al of the BCL2 family of proteins. In a preferred embodiment, the inhibitor at least targets BCL-XL. In an alternate embodiment, BCL2 is not the only BCL2 family member the inhibitor targets (i.e. the inhibitor is not BCL2 specific).
  • the BCL2 family inhibitor is selected from, but not necessarily limited to, the group consisting of a BH3 mimetic agent, an RNAi against a BCL2 family member, and an antibody against a BCL2 family member.
  • the BCL2 family inhibitor is a BH3 mimetic agent.
  • the BH3 mimetic agent used in any aspect described herein is a BH3 mimetic agent that inhibits BCL-XL. In one embodiment the BH3 mimetic agent is a selective inhibitor of BCL-XL. In some embodiments the BH3 mimetic agent in any aspect described herein is a peptide. In some embodiments the BH3 mimetic agent in any aspect described herein is a small molecule compound ("BH3 mimetic compound"). In some embodiments the BH3 mimetic compound is a compound having a structure selected from the group consisting of Formulas I-XII, as set forth herein.
  • a BCL2 family inhibitor for the preparation of a medicament for treating an infection.
  • BCL2 family inhibitor for use in the treatment of an infection.
  • composition containing a therapeutically effective amount of a BCL2 family inhibitor and a bactericidal or bacteriostatic agent.
  • kits when used for the treatment of a bacterial infection comprising a BCL2 family inhibitor.
  • the kit includes, in addition to the BCL2 family inhibitor, a bactericidal or bacteriostatic agent.
  • composition of matter, group of steps or group of compositions of matter shall be taken to encompass one and a plurality (i.e. one or more) of those steps, compositions of matter, groups of steps or group of compositions of matter.
  • Figure 1 - ABT-737 restricts Legionella burdens in BMDMs, in vitro, by inducing host cell death, a, b, c, Draq7 positive (dead) bone marrow-derived macrophages (BMDMs) were quantified by live-cell imaging over 72 h, following infection with (a, c) AflaA or (b) AdotA Legionella at (a, b) MOI of 10 or (c) 25, and addition of 10 ⁇ ABT-737. Mean and standard deviation (SD) of three repeats are shown.
  • SD standard deviation
  • Figure 2 Targeting BCL-XL, rather than BCL-2, limits intracellular Legionella loads, a
  • Right schematic depicting the pro-survival BCL-2 family members targeted by the different BH3-mimetics used in this study (A-l 155463 is abbreviated to 463).
  • CFUs Bacterial burden
  • WT wild type
  • f g, Live-cell microscopic analysis of Draq7 positive (dead) BMDMs infected with AflaA and AdotA. Mean and SD of three repeats are shown. Data representative of at least three independent experiments.
  • Figure 3 Loss of BCL-XL induces apoptosis in Legionella-inf ct d macrophages.
  • a Time course immuno-blot analysis for cleaved (i.e., activated) caspase-3 in wild type (WT) and BCL-XL-deficient BMDMs infected with AflaA or AdotA or b, A-l 155463 (463)-treated WT BMDMs infected with AflaA. Actin blots are loading controls.
  • FIG. 5 - ABT-737 protects BMDMs from L. pneumophila-inductd cell death, a, b, Draq7 positive (dead) bone marrow-derived macrophages (BMDMs) were quantified over 72 h using time-lapse imaging in the presence of 10 ⁇ ABT-737 or vehicle (DMSO) (b) without infection or (a) after infection with wild type L. pneumophila, at an MOI of 10. Mean and SD are shown. Data representative of at least three independent experiments.
  • FIG. 6 - BCL-XL protects BMDMs from wild type L. pneumophila-inductd death
  • a Immuno-blot analysis of BCL-XL protein content in bone marrow derived macrophages (BMDMs) derived from Bcl- ⁇ ⁇ 0X ;ER-Cre mice, whereby progenitor cells were left untreated, or else treated with 4-hydroxytamoxifen (4-HT; 50 nM, 2 days post-harvesting) to induce deletion of the two floxed Bcl-x alleles
  • BMDMs bone marrow derived macrophages
  • 4-HT 4-hydroxytamoxifen
  • 50 nM 4 days post-harvesting
  • b Live-cell microscopic analysis of Draq7 positive (dead) WT (C57BL/6) and BCL-XL-deficient BMDMs infected with WT L. pneumophila. Mean and SD are shown. Data representative of at least three independent experiments.
  • Dotted line shows infection level at 6 h.
  • FIG. 9 - A BH3 mimetic having the structure of Formula (XIII) targets BCL-XL and inhibits replication of Legionella longbeachae in vivo.
  • FIG 10 - A BCL-XL-Selective BH3 Mimetic Compound, A-1331852 (Formula XIII) is Effective in Treatment of a Model of Latent Mycobacterium tuberculosis infection, a, Schematic overview of an experiment in which mice, in a latent model of M. tuberculosis infection, were administered either vehicle or BCL-XL-selective BH3 mimetic compound, A-1331852 (Formula XIII).
  • the term about refers to +/- 10%, more preferably +/- 5%, of the designated value.
  • the term "or” is intended to mean an inclusive “or” rather than an exclusive “or.” That is, unless specified otherwise, or clear from context, "X employs A or B” is intended to mean any of the natural inclusive permutations. That is, if X employs A; X employs B; or X employs both A and B, then "X employs A or B” is satisfied under any of the foregoing instances. Further, at least one of A and B and/or the like generally means A or B or both A and B.
  • the articles “a” and “an” as used in this application and the appended claims may generally be construed to mean “one or more” unless specified otherwise or clear from context to be directed to a singular form.
  • acute infection refers to an infection characterized by rapid onset of disease, a relatively brief period of symptoms, and resolution within days. It is usually accompanied by early production of the underlying infectious pathogen and elimination of infection by the host immune system.
  • subacute refers to an infection that is not chronic and that runs a rapid and severe, but less than acute, course.
  • anti-infective agent refers to any agent (e.g., a small molecule compound) that directly inhibits the ability of an infectious pathogen to infect, replicate, survive, or otherwise cause damage to its host.
  • anti- infective agents include, but are not limited to, antibiotics, antiviral drugs, antifungal drugs, and antiprotozoal drugs.
  • BCL2 family inhibitors, as used herein, are specifically excluded from being considered anti-infective agents.
  • BCL2 family inhibitor refers to any agent that inhibits the pro-survival activity of at least one member of the BCL2 protein family subgroup that includes BCL-2, BCL-XL, BCL-W, MCL-1, and Al/BFLl .
  • types of BCL2 inhibitors include, but are not limited to, small molecule compound BH3 mimetics, peptides, RNAi, and antibodies.
  • BH3 mimetic agent refers to any agent (e.g., a small molecule compound or a peptide) that interacts, in a manner analogous to a BH3 domain, with the hydrophobic groove of the prosurvival BCL-2 proteins (BCL-2, BCL-XL, BCL-W, MCL-1, and Al/BFLl) to antagonize their prosurvival (antiapoptotic) activity.
  • BCL-2, BCL-XL, BCL-W, MCL-1, and Al/BFLl prosurvival BCL-2 proteins
  • Examples of BH3 only domain proteins include NOXA, BAD, and BIM.
  • co-administration are meant to encompass administration of the selected therapeutic agents to a single patient, and are intended to include treatment regimens in which the agents are administered by the same or different route of administration or at the same or different time.
  • an “effective amount” or “therapeutically effective amount,” as used herein, refer to a sufficient amount of an agent or a compound being administered which will relieve to some extent one or more of the symptoms of the disease or condition being treated. The result can be reduction and/or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system.
  • an “effective amount” for therapeutic uses is the amount of the composition including a compound as disclosed herein required to provide a clinically significant decrease in disease symptoms without undue adverse side effects.
  • An appropriate “effective amount” in any individual case may be determined using techniques, such as a dose escalation study.
  • the term "therapeutically effective amount” includes, for example, a prophylactically effective amount.
  • an “effective amount” of a therapeutic agent disclosed herein is an amount effective to achieve a desired pharmacologic effect or therapeutic improvement without undue adverse side effects. It is understood that “an effect amount” or “a therapeutically effective amount” can vary from subject to subject, due to variation in metabolism of the compound of any of age, weight, general condition of the subject, the condition being treated, the severity of the condition being treated, and the judgment of the prescribing physician. By way of example only, therapeutically effective amounts may be determined by routine experimentation, including but not limited to a dose escalation clinical trial.
  • a "therapeutically effective amount" of each therapeutic agent can refer to an amount of the therapeutic agent that would be therapeutically effective when used on its own, or may refer to a reduced amount that is therapeutically effective by virtue of its combination with one or more additional therapeutic agents.
  • inhibitors in reference to an inhibitor indicates that inhibition of the activity of one member of a protein family by an inhibitor may be significantly higher (e.g., 25%, 50%, 300%, 1000%) higher) than for another member of the same protein family, but does not necessarily mean that the inhibitor exclusively inhibits one and only one member of a protein family.
  • small molecule refers to a chemical compounds or molecule having a molecular weight below 2000 daltons.
  • treating refers to both direct treatment of a subject by a medical professional (e.g., by administering a therapeutic agent to the subject), or indirect treatment, effected, by at least one party, (e.g., a medical doctor, a nurse, a pharmacist, or a pharmaceutical sales representative) by providing instructions, in any form, that (i) instruct a subject to self-treat according to a claimed method (e.g., self-administer a drug) or (ii) instruct a third party to treat a subject according to a claimed method.
  • a medical professional e.g., by administering a therapeutic agent to the subject
  • indirect treatment effected, by at least one party, (e.g., a medical doctor, a nurse, a pharmacist, or a pharmaceutical sales representative) by providing instructions, in any form, that (i) instruct a subject to self-treat according to a claimed method (e.g., self-administer a drug) or (ii) instruct a third party to treat
  • treating or “treatment” are prevention or reduction of the disease to be treated, e.g., by administering a therapeutic at a sufficiently early phase of disease to prevent or slow its progression.
  • the methods described herein include treating an infection by administering a therapeutically effective amount a BCL2 family inhibitor, e.g., a BH3 mimetic agent.
  • a BCL2 family inhibitor e.g., a BH3 mimetic agent.
  • infectious pathogens upon infection of a host cell, upregulate the pro- survival activity of one or more proteins in the BCL2 family to prevent the host cell apoptosis, which would otherwise curtail the infectious cycle.
  • a BCL2 family inhibitor restores apoptosis in such cells thereby blocking the ability of certain pathogens to replicate and infect other cells.
  • the infection to be treated is a bacterial infection.
  • bacterial infections to be treated by the methods described herein include, but are not limited to, infectious caused by one or more of Legionella (e.g., L. pneumophila and L. longbeachae), Mycobacterium, Salmonella, and Pseudomonas.
  • Legionella e.g., L. pneumophila and L. longbeachae
  • Mycobacterium e.g., Mycobacterium, Salmonella, and Pseudomonas.
  • Salmonella e.g., S. pneumophila and L. longbeachae
  • Pseudomonas e.g., Pseudomonas.
  • the subject may also be administered a bactericidal or bacteriostatic agent.
  • bactericidal agents include, but are not limited to, antibiotics such as aminoglycosides, ansamycins, carbapenems, cephalosporins, glycopeptides, lipopeptides, monobactams, nitrofurans, penicillins, quinolones, capreomycine, cycloserine, ethionamide, isoniazid, pyrazinamide, rifamycin, rifabutin, rifapentine, and streptomycin.
  • antibiotics such as aminoglycosides, ansamycins, carbapenems, cephalosporins, glycopeptides, lipopeptides, monobactams, nitrofurans, penicillins, quinolones, capreomycine, cycloserine, ethionamide, isoniazid, pyrazinamide, rifamycin, rifabutin, rifapentine, and streptomycin.
  • bacteriostatic agents include, but are not limited to, bacteriostatic antibiotics such as lincosamides, macrolides, sulphonamides, tetracyclines, spectinomycin, trimethoprim, clindamycin, ethambutol, novobiocin, tigecycline, and, oxazolidinones.
  • bacteriostatic antibiotics such as lincosamides, macrolides, sulphonamides, tetracyclines, spectinomycin, trimethoprim, clindamycin, ethambutol, novobiocin, tigecycline, and, oxazolidinones.
  • the bacterial infection to be treated includes infection by antibiotic-resistant bacteria, e.g., antibiotic resistant forms of any of Staphylococcus aureus, Enterococcus, and Pseudomonas aeruginosa, and Achinobacter baumannii.
  • antibiotic-resistant bacteria e.g., antibiotic resistant forms of any of Staphylococcus aureus, Enterococcus, and Pseudomonas aeruginosa, and Achinobacter baumannii.
  • the infection to be treated is a viral infection other than an infection by an influenza virus.
  • viral infections to be treated by the methods described herein include, but are not limited to, infections by HSV, HTLV-1, HTLV-2, HIV, a respiratory virus, rhinovirus, or a coronavirus, or ebola
  • the infection to be treated is a fungal infection.
  • infections include, but are not limited to, yeast infections ⁇ e.g., Candida infections), Aspergillus, Pneumocystitis, and Cryptococcus.
  • the infection to be treated includes a protozoal infection.
  • protozoal infections to be treated include, but are not limited to, infections by Leishmania, Trypanosomoa cruzi, Toxoplasma gondii, and Encephalitozoon .
  • the administration occurs within a period ranging from about 30 minutes to about 72 hours following a diagnosis of the infection, e.g., about 45 minutes, 1 hours, 2 hours, 3 hours 6 hours, 8 hours, 12 hours, 18 hours, 24 hours, 36 hours, 50 hours, 60 hours, 65 hours, or another time period from about 30 minutes to about 72 hours following diagnosis of the infection.
  • the administration occurs within a period from about 6 hours to ab out 48 hours following a diagnosis of the infection.
  • administration of the BCL-2 family inhibitor occurs no later than about 30 minutes to about 60 hours following a diagnosis of the infection, e.g., within about 45 minutes, 1 hours, 2 hours, 3 hours 6 hours, 8 hours, 12 hours, 18 hours, 24 hours, 36 hours, 48 hours, 50 hours, or another time period no later than about 30 minutes to about 60 hours following diagnosis of the infection. In one embodiment the administration occurs no later than about 48 hours following a diagnosis of the infection.
  • one or more cell types become infected by the infectious pathogen.
  • the subject to be treated is suffering from an infection that includes infection of macrophages.
  • the infected macrophages are alveolar macrophages.
  • the subject to be treated as per any of the methods described herein is a subject that was not diagnosed as suffering from a cancer.
  • the subject to be treated is suffering from an acute or subacute infection.
  • the subject to be treated is suffering from tuberculosis, pneumonia, or an acute respiratory infection.
  • Subjects that can be treated by the methods described herein include, but are not limited to, humans, non-human primates, pigs, cows, sheep, ducks, and chicken.
  • the subject to be treated is a human patient.
  • a BCL2 family inhibitor is selected from among a BH3 mimetic agent, an RNAi against a BCL2 family member, and an antibody against a BCL2 family member.
  • a BH3 mimetic agent suitable for treatment in a method described herein is a BH3 mimetic agent that inhibits the anti-apoptotic activity of BCL-XL, but can also inhibit the anti-apoptotic activity of other BCL-2 proteins, e.g., BCL-2 and BCL-W.
  • the BH3 mimetic agent to be used is a BCL-XL-selective inhibitor.
  • the BH3 mimetic agent inhibits BCL-2, BCL-W, and BCL-XL.
  • Suitable BH3 mimetic agents include, but are not limited to, small molecules, peptide mimetics (e.g., terphenyl peptide mimetics), and peptides.
  • a BH3 mimetic agent is a peptide, e.g., a stapled BH3 peptide as described in, et al., Labelle et al. (2012); and Walensky et al. (2004).
  • the BH3 mimetic agent to be used is a BH3 mimetic compound.
  • Suitable BH3 mimetic agent compounds for the methods described herein include, but are not limited to, those disclosed in international patent publications WO2010080478, WO2005049594, WO2007040650, WO2010080503,
  • a suitable BH3 mimetic compound is N-(4-(4-((4'- chloro-(l,-biphenyl)-2-yl)methyl)piperazin-l-yl)-benzoyl)-4-(((lR)-3-(dimethylamino)-l- ((phenylsulfanyl)methyl)propyl)arnino)-3 -nitrobenzene-sulfonamide, otherwise known as ABT-737, the structure of which is shown below as Formula (I), or an analogue or derivative thereof:
  • the BH3 mimetic compound to be used is N-(4-(4-((2- (4- chiorophenyl)-5,5-dimethyf-l-cyclohex-l-en-f-y ⁇
  • the BH3 mimetic compound to be used is a BCL-XL- selective inhibitor compound A-1155463, having the structure of Formula (III) , or an analogue or derivative thereof:
  • the BH3 mimetic compound to be used is A-385358, having the structure of Formula (IV), or an analogue or derivative thereof:
  • the BH3 mimetic compound to be used is BM-957, having the structure of Formula (V), or an analogue or derivative thereof:
  • the BH3 mimetic compound to be used has the structure of Formula (VI), or an analogue or derivative thereof:
  • the BH3 mimetic compound to be used has the structure of Formula (VII), or an analogue or derivative thereof:
  • the BH3 mimetic compound to be used is MIM-1, having the structure of Formula (VIII), or an analogue or derivative thereof:
  • the BH3 mimetic compound to be used has the structure of Formula (IX), or an analogue or derivative thereof:
  • the BH3 mimetic compound to be used is BH3-M6, having the structure of Formula (X), or an analogue or derivative thereof:
  • the BH3 compound to be used is WEHI-539, having the structure of Formula (XI), or an analogue or derivative thereof:
  • the BH3 compound to be used is a compound having the structure of Formula (XII), or an analogue or derivative thereof:
  • the BH3 compound to be used is the compound disclosed in example 3 of international patent publication WO2013055897and having the structure of Formula (XIII), or an analogue or derivative thereof:
  • RNA Interference RNA Interference
  • the BCL2 family inhibitor includes one or more RNAis against a BCL2 family inhibitor.
  • the RNAi is directed against BCL-XL.
  • RNA interference refer generally to a process in which a double-stranded RNA molecule reduces the expression of a nucleic acid sequence with which the double-stranded RNA molecule shares substantial or total homology.
  • RNA interference can be achieved using non-RNA double stranded molecules (see, for example, US 20070004667).
  • a BCL2 family inhibitor comprises nucleic acid molecules comprising and/or encoding double-stranded regions for RNA interference.
  • the nucleic acid molecules are typically RNA but may comprise chemically-modified nucleotides and non-nucleotides.
  • the double-stranded regions should be at least 19 contiguous nucleotides, for example about 19 to 23 nucleotides, or may be longer, for example 30 or 50 nucleotides, or 100 nucleotides or more.
  • the full-length sequence corresponding to the entire gene transcript may be used. Preferably, they are about 19 to about 23 nucleotides in length.
  • the degree of identity of a double-stranded region of a nucleic acid molecule to the targeted transcript should be at least 90% and more preferably 95-100%.
  • the nucleic acid molecule may of course comprise unrelated sequences which may function to stabilize the molecule.
  • short interfering RNA or "siRNA” as used herein refers to a nucleic acid molecule which comprises ribonucleotides capable of inhibiting or down regulating gene expression, for example by mediating RNAi in a sequence-specific manner, wherein the double stranded portion is less than 50 nucleotides in length, preferably about 19 to about 23 nucleotides in length.
  • the siRNA can be a nucleic acid molecule comprising self-complementary sense and antisense regions, wherein the antisense region comprises nucleotide sequence that is complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense region having nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof.
  • the siRNA can be assembled from two separate oligonucleotides, where one strand is the sense strand and the other is the antisense strand, wherein the antisense and sense strands are self-complementary.
  • siRNA is meant to be equivalent to other terms used to describe nucleic acid molecules that are capable of mediating sequence specific RNAi, for example micro-RNA (miRNA), short hairpin RNA (shRNA), short interfering oligonucleotide, short interfering nucleic acid (siNA), short interfering modified oligonucleotide, chemically-modified siRNA, post-transcriptional gene silencing RNA (ptgsRNA), and others.
  • miRNA micro-RNA
  • shRNA short hairpin RNA
  • siNA short interfering nucleic acid
  • ptgsRNA post-transcriptional gene silencing RNA
  • RNAi is meant to be equivalent to other terms used to describe sequence specific RNA interference, such as post transcriptional gene silencing, translational inhibition, or epigenetics.
  • siRNA molecules as described herein can be used to epigenetically silence genes at both the post-transcriptional level or the pre-transcriptional level.
  • epigenetic regulation of gene expression by siRNA molecules as described herein can result from siRNA mediated modification of chromatin structure to alter gene expression.
  • RNA short-hairpin RNA
  • short-hairpin RNA an RNA molecule where less than about 50 nucleotides, preferably about 19 to about 23 nucleotides, is base paired with a complementary sequence located on the same RNA molecule, and where said sequence and complementary sequence are separated by an unpaired region of at least about 4 to about 15 nucleotides which forms a single- stranded loop above the stem structure created by the two regions of base complementarity.
  • shRNAs are dual or bi-finger and multi-finger hairpin dsRNAs, in which the RNA molecule comprises two or more of such stem-loop structures separated by single-stranded spacer regions.
  • nucleic acid molecules comprising a double-stranded region can be generated by any method known in the art, for example, by in vitro transcription, recombinantly, or by synthetic means.
  • nucleic acid molecules and “double-stranded RNA molecule” includes synthetically modified bases such as, but not limited to, inosine, xanthine,
  • hypoxanthine 2-aminoadenine, 6-methyl-, 2-propyl- and other alkyl- adenines, 5-halo uracil, 5-halo cytosine, 6-aza cytosine and 6-aza thymine, pseudo uracil, 4-thiuracil, 8- halo adenine, 8-aminoadenine, 8-thiol adenine, 8-thiolalkyl adenines, 8-hydroxyl adenine and other 8-substituted adenines, 8-halo guanines, 8-amino guanine, 8-thiol guanine, 8-thioalkyl guanines, 8-hydroxyl guanine and other substituted guanines, other aza and deaza adenines, other aza and deaza guanines, 5-trifluoromethyl uracil and 5- trifluoro cytosine.
  • RNA is used to target various BCL2 family members are known in the art, as exemplified without limitation, for BCL-XL, in Tsai et al. (2014); for BCL2 and BCL- W in Crawford et al. (2010); for MCL-1 in Keuling et al. (2009); and for A1/BFL1 in Ottina et al. (2012).
  • a BCL2 family inhibitor is an antibody against one or more BCL2 family members.
  • the antibody inhibits BCL-XL as well at least one other BCL2 family member.
  • the antibody preferentially inhibits BCL-XL.
  • the antibody is an antibody modified to penetrate or be taken up (passively or actively) in mammalian cells.
  • antibody includes polyclonal antibodies, monoclonal antibodies, bispecific antibodies, fusion diabodies, triabodies, heteroconjugate antibodies, chimeric antibodies including intact molecules as well as fragments thereof, and other antibody-like molecules.
  • Antibodies include modifications in a variety of forms including, for example, but not limited to, domain antibodies including either the VH or VL domain, a dimer of the heavy chain variable region (VHH, as described for a camelid), a dimer of the light chain variable region (VLL), Fv fragments containing only the light (VL) and heavy chain (VH) variable regions which may be joined directly or through a linker, or Fd fragments containing the heavy chain variable region and the CHI domain.
  • domain antibodies including either the VH or VL domain, a dimer of the heavy chain variable region (VHH, as described for a camelid), a dimer of the light chain variable region (VLL), Fv fragments containing only the light (VL) and heavy chain (VH) variable regions which may be joined
  • a scFv consisting of the variable regions of the heavy and light chains linked together to form a single-chain antibody (Bird et al., 1988; Huston et al., 1988) and oligomers of scFvs such as diabodies and triabodies are also encompassed by the term "antibody”. Also encompassed are fragments of antibodies such as Fab, (Fab')2 and FabFc2 fragments which contain the variable regions and parts of the constant regions. Complementarity determining region (CDR)-grafted antibody fragments and oligomers of antibody fragments are also encompassed.
  • the heavy and light chain components of an Fv may be derived from the same antibody or different antibodies thereby producing a chimeric Fv region.
  • the antibody may be of animal (for example mouse, rabbit or rat) or human origin or may be chimeric (Morrison et al., 1984) or humanized (Jones et al., 1986).
  • the term "antibody” includes these various forms. Using the guidelines provided herein and those methods well known to those skilled in the art which are described in the references cited above and in such publications as Harlow & Lane, Antibodies: a Laboratory Manual, Cold Spring Harbor Laboratory, (1988) the antibodies for use in the methods of the present invention can be readily made.
  • the antibodies may be Fv regions comprising a variable light (VL) and a variable heavy (VH) chain in which the light and heavy chains may be joined directly or through a linker.
  • VL variable light
  • VH variable heavy chain
  • a linker refers to a molecule that is covalently linked to the light and heavy chain and provides enough spacing and flexibility between the two chains such that they are able to achieve a conformation in which they are capable of specifically binding the epitope to which they are directed.
  • Protein linkers are particularly preferred as they may be expressed as an intrinsic component of the Ig portion of the fusion polypeptide.
  • recombinantly produced single chain scFv antibody preferably a humanized scFv
  • scFv antibody preferably a humanized scFv
  • the antibodies have the capacity for intracellular transmission.
  • Antibodies which have the capacity for intracellular transmission include antibodies such as camelids and llama antibodies, shark antibodies (IgNARs), scFv antibodies, intrabodies or nanobodies, for example, scFv intrabodies and VHH intrabodies.
  • Such antigen binding agents can be made as described by Harmsen and De Haard (2007), Tibary et al. (2007), and Muyldermans (2001), and references cited therein.
  • Yeast SPLINT antibody libraries are available for testing for intrabodies which are able to disrupt protein-protein interactions (see for example, Visintin et al. (2008) for methods for their production).
  • Such agents may comprise a cell-penetrating peptide sequence or nuclear-localizing peptide sequence such as those disclosed in Constantini et al. (2008). Also useful for in vivo delivery are Vectocell or Diato peptide vectors such as those disclosed in De Coupade et al. (2005) and Meyer-Losic et al. (2006).
  • the antibodies may be fused to a cell penetrating agent, for example a cell-penetrating peptide.
  • Cell penetrating peptides include Tat peptides, Penetratin, short amphipathic peptides such as those from the Pep-and MPG-families, oligoarginine and oligolysine.
  • the cell penetrating peptide is also conjugated to a lipid (C6-C18 fatty acid) domain to improve intracellular delivery (Koppelhus et al., 2008). Examples of cell penetrating peptides can be found in Howl et al., (2007) and Deshayes et al. (2008).
  • the invention also provides the therapeutic use of antibodies fused via a covalent bond (e.g. a peptide bond), at optionally the N-terminus or the C-terminus, to a cell-penetrating peptide sequence.
  • Antibodies which inhibit one or more of BCL-XL, BCL-2, BCL-W, MCL-1, or
  • Al/BFL-1 activity are available from various sources such as Santa Cruz Biotechnology, and as exemplified for BCL-2 in Cohen-Saidon et al. (2003).
  • the antibody to be used is an antibody that inhibits the activity of BCL- XL.
  • the antibody to be used is antibody that preferentially inhibits BCL-XL.
  • a method for treating a subject suffering from an infection includes administration of a pharmaceutical composition containing at least one BCL2 family inhibitor, or a pharmaceutically acceptable salt, pharmaceutically acceptable N-oxide, pharmaceutically active metabolite, pharmaceutically acceptable prodrug, or pharmaceutically acceptable solvate thereof, in therapeutically effective amounts to said subject.
  • Treatment can be for prophylactic and/or therapeutic treatments.
  • a BCL2 family inhibitor e.g., a BH3 mimetic compound
  • compositions containing a BCL2 family inhibitor are administered to a patient susceptible to or otherwise at risk of infection, for example, during or after recent travel to an epidemic zone.
  • a patient susceptible to or otherwise at risk of infection for example, during or after recent travel to an epidemic zone.
  • Such an amount is defined to be a "prophylactically effective amount or dose,” i.e., a dose sufficient to prevent or reduce the onset of infection.
  • prophylactically effective amounts i.e., a dose sufficient to prevent or reduce the onset of infection.
  • the precise amounts also depend on the patient's state of health, weight, timing, etc. It is considered well within the skill of the art for one to determine such prophylactically effective amounts by routine experimentation (e.g., a dose escalation clinical trial).
  • the administration of a BCL2 family inhibitor may be given continuously; alternatively, the dose of drug being administered may be temporarily reduced or temporarily suspended for a certain length of time (i.e., a "drug holiday").
  • the length of the drug holiday can vary between 2 days and 1 year, including by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, or 60 days.
  • the dose reduction during a drug holiday may be from 10%- 100%, including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.
  • the amount of a given BCL2 family inhibitor that will be suitable as a therapeutically effective dose will vary depending upon factors such as the particular BCL-2 family inhibitor, infection and its severity, the characteristics (e.g., weight) of the subject or host in need of treatment, and the properties of the infectious pathogen (e.g., its doubling time and drug resistance), but can nevertheless be routinely determined in a manner known in the art according to the particular circumstances surrounding the case, including, e.g., the specific agent being administered, the route of administration, the condition being treated, and the subject or host being treated. In general, however, doses employed for adult human treatment will typically be in the range of 0.02-5000 mg per day, or from about 1-1500 mg per day. The desired dose may conveniently be presented in a single dose or as divided doses administered simultaneously (or over a short period of time) or at appropriate intervals, for example as two, three, four or more sub-doses per day.
  • Toxicity and therapeutic efficacy of such therapeutic regimens can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, including, but not limited to, the determination of the LD 50 (the dose lethal to 50% of the population) and the ED 50 (the dose therapeutically effective in 50% of the population).
  • the dose ratio between the toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio between LD 50 and ED 50 .
  • BCL2 family inhibitors, and particularly BH3 mimetic compounds, exhibiting high therapeutic indices are preferred.
  • the data obtained from cell culture assays and animal studies can be used in formulating a range of dosage for use in human and non- human subjects.
  • the dosage of such compounds lies preferably within a range of circulating concentrations that include the ED 50 with minimal toxicity.
  • the dosage may vary within this range depending upon the dosage form employed and the route of administration utilized.
  • BCL2 family inhibitors can also be used in combination with other agents used to treat infectious diseases (anti-infective agents), that are selected for their therapeutic value for the infection to be treated.
  • anti-infective agents that are selected for their therapeutic value for the infection to be treated.
  • the compositions described herein and, in embodiments where combinational therapy is employed other agents do not necessarily have to be administered in the same pharmaceutical composition, and may, because of different physical and chemical characteristics, preferably be administered by different routes.
  • the determination of the mode of administration and the advisability of administration, where possible, in the same pharmaceutical composition is well within the knowledge of the skilled clinician.
  • the initial administration can be made according to established protocols known in the art, and then, based upon the observed effects, the dosage, modes of administration and times of administration can be modified by the skilled clinician.
  • a BCL2 family inhibitor and an additional therapeutic agent may be administered concurrently (e.g., simultaneously, essentially simultaneously or within the same treatment protocol) or sequentially, depending upon the nature and phase of the infection, the condition of the patient, and the actual choice of therapeutic agents used.
  • the determination of the order of administration, and the number of repetitions of administration of each therapeutic agent during a treatment protocol, is well within the knowledge of the skilled physician after evaluation of the disease being treated and the condition of the patient.
  • therapeutically-effective dosages can vary when the drugs are used in treatment combinations.
  • Methods for experimentally determining therapeutically-effective dosages of drugs and other agents for use in combination treatment regimens are described in the literature.
  • metronomic dosing i.e., providing more frequent, lower doses in order to minimize toxic side effects
  • Combination treatment further includes periodic treatments that start and stop at various times to assist with the clinical management of the patient.
  • dosages of co-administered therapeutic agents will of course vary depending on the type of co-agents employed, on the specific BCL2 family inhibitor, on the infection being treated and so forth
  • the dosage regimen to treat, prevent, or ameliorate the condition(s) for which relief is sought can be modified in accordance with a variety of factors. These factors include the disorder from which the subject suffers, as well as the age, weight, sex, diet, and medical condition of the subject. Thus, the dosage regimen actually employed can vary widely and therefore can deviate from the dosage regimens set forth herein.
  • the BCL2 family inhibitor and additional therapeutic agent which make up a combination therapy disclosed herein may be a combined dosage form or in separate dosage forms intended for substantially simultaneous administration.
  • the pharmaceutical agents that make up the combination therapy may also be administered sequentially, with either therapeutic compound being administered by a regimen calling for two-step administration.
  • the two-step administration regimen may call for sequential administration of the active agents or spaced-apart administration of the separate active agents.
  • the time period between the multiple administration steps may range from, a few minutes to several hours, depending upon the properties of each pharmaceutical agent, such as potency, solubility, bioavailability, plasma half-life and kinetic profile of the pharmaceutical agent. Circadian variation of various physiological parameters may also be evaluated to determine the optimal dose interval.
  • administering may be used in combination with procedures that may provide additional or synergistic benefit to the patient.
  • patients may undergo genetic testing to identify genetic variation in their own genome or a pathogen's genome so as to optimize treatment parameters, e.g., the type of BCL2 family inhibitor to be administered, dosing regimen, and co-administration with anti- infective agents.
  • Initial administration can be via any route practical, such as, for example, an intravenous injection, a bolus injection, infusion over 5 minutes to about 5 hours, a pill, a capsule, inhaler, injection, transdermal patch, buccal delivery, and the like, or combination thereof.
  • a compound should be administered as soon as is practicable after the onset of a disease or condition is detected or suspected, and for a length of time necessary for the treatment of the infection.
  • a BCL2 family inhibitor can be used in combination with one or more bactericidal or bacteriostatic agents, including, but not limited to: gentamicin, tobramycin, streptomycin, doripenem, cefazolin, cefaclor, cefepime, ceftobiprole, vancomycin, oritavancin, clindamycin, daptomycin, azithromycin, telithromycin, furazolidone, linezolid, amoxicillin, amoxicillin/clavulanate, ciprofloxacin, gemifloxacin, sulfacetamide, minocycline, capreomycin, cycloserine, ethambutol, ethionamide, isoniazid, pyrazinamide, rifampicin, and rifabutin.
  • bactericidal or bacteriostatic agents including, but not limited to: gentamicin, tobramycin, streptomycin, doripenem, ce
  • a BCL2 family inhibitor can be used in combination with one or more anti-viral drugs, including, but not limited to: cidofovir, ribavirin, acyclovir, ganciclovir, vidarabine, abacavir, nevirapine, ritonavir, gemcitabine, and decitabine.
  • a BCL2 family inhibitor can be used in combination with one or more antifungal drugs including, but not limited to: candicidin, fluconazole, tioconazole, voriconazole, abafungin, terbinafine, micafungin, and undecylenic acid.
  • a BCL-2 family inhibitor can be used in combination with one or more anti protozoal drugs including, but not limited to: ambisome, amphotericin, lumieririne, chloroquine, daraprim, diloxanide furoate, doxycycline monohydrate, lariam, malarone, pentamidine isetionate, pentostam, proquanil hydrochloride, tinidazole, and wellvone.
  • anti protozoal drugs including, but not limited to: ambisome, amphotericin, lumieririne, chloroquine, daraprim, diloxanide furoate, doxycycline monohydrate, lariam, malarone, pentamidine isetionate, pentostam, proquanil hydrochloride, tinidazole, and wellvone.
  • compositions comprising therapeutically effective amounts of (i) a BCL2 family inhibitor, including, but not limited to, any BCL2 family inhibitor described herein; and (ii) an anti-infective agent.
  • the BCL2 family inhibitor is a BH3 mimetic agent, an RNAi against a BCL2 family member, or an antibody against a BCL2 family member.
  • the pharmaceutical composition comprises a BH3 mimetic agent, including, but not limited to any BH3 mimetic agent described herein.
  • the BH3 mimetic agent in the pharmaceutical composition preferentially inhibits BCL-XL relative to other BCL2 family members.
  • a pharmaceutical composition comprises a BCL2 family inhibitor and a bactericidal or bacteriostatic agent, including, but not limited to any bactericidal or bacteriostatic agent recited herein.
  • the pharmaceutical composition comprises a BCL2 family inhibitor and an anti-viral agent, including, but not limited to, any antiviral agent recited herein.
  • the pharmaceutical composition comprises a BCL2 family inhibitor and an antifungal agent including, but not limited to, any antifungal agent recited herein.
  • the pharmaceutical composition comprises a BCL2 family inhibitor and an antiprotozoal agent including, but not limited to, any antiprotozoal agent recited herein.
  • compositions described herein can be formulated for administration to a subject via any conventional means including, but not limited to, oral, parenteral ⁇ e.g., intravenous, subcutaneous, or intramuscular), buccal, inhalation, intranasal, rectal or transdermal administration routes.
  • oral parenteral ⁇ e.g., intravenous, subcutaneous, or intramuscular
  • buccal inhalation
  • intranasal rectal or transdermal administration routes.
  • compositions described herein which include a BCL2 family inhibitor ⁇ e.g., a BH3 mimetic compound) and an anti-infective agent, can be formulated into any suitable dosage form, including but not limited to, aqueous oral dispersions, liquids, mists, gels, syrups, elixirs, slurries, suspensions and the like, for oral ingestion by a patient to be treated, solid oral dosage forms, aerosols, controlled release formulations, fast melt formulations, effervescent formulations, lyophilized formulations, tablets, powders, pills, dragees, capsules, delayed release formulations, extended release formulations, pulsatile release formulations, multiparticulate formulations, and mixed immediate release and controlled release formulations.
  • aqueous oral dispersions liquids, mists, gels, syrups, elixirs, slurries, suspensions and the like
  • solid oral dosage forms aerosols, controlled release formulations, fast melt formulations,
  • compositions for oral use can be obtained by mixing one or more solid excipient with one or more of the compounds described herein, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores.
  • Suitable excipients include, for example, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or others such as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate.
  • disintegrating agents may be added, such as the cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.
  • compositions which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol.
  • the push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers.
  • the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols.
  • stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration.
  • the solid dosage forms disclosed herein may be in the form of a tablet, (including a suspension tablet, a fast-melt tablet, a bite-disintegration tablet, a rapid-disintegration tablet, an effervescent tablet, or a caplet), a pill, a powder (including a sterile packaged powder, a dispensable powder, or an effervescent powder) a capsule (including both soft or hard capsules, e.g., capsules made from animal- derived gelatin or plant-derived HPMC, or "sprinkle capsules"), solid dispersion, solid solution, bioerodible dosage form, controlled release formulations, pulsatile release dosage forms, multiparticulate dosage forms, pellets, granules, or an aerosol.
  • a tablet including a suspension tablet, a fast-melt tablet, a bite-disintegration tablet, a rapid-disintegration tablet, an effervescent tablet, or a caplet
  • a pill including a ster
  • the pharmaceutical formulation is in the form of a powder. In still other embodiments, the pharmaceutical formulation is in the form of a tablet, including but not limited to, a fast-melt tablet. Additionally, pharmaceutical formulations described herein may be administered as a single capsule or in multiple capsule dosage form. In some embodiments, the pharmaceutical formulation is administered in two, or three, or four, capsules or tablets.
  • solid dosage forms e.g., tablets, effervescent tablets, and capsules
  • solid dosage forms are prepared by mixing particles of a BCL2 family inhibitor and an anti- infective compound with one or more pharmaceutical excipients to form a bulk blend composition.
  • these bulk blend compositions as homogeneous, it is meant that the particles of the active agents, are dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms, such as tablets, pills, and capsules.
  • the individual unit dosages may also include film coatings, which disintegrate upon oral ingestion or upon contact with diluent. These formulations can be manufactured by conventional pharmacological techniques.
  • the pharmaceutical solid dosage forms described herein can include a BCL2 family inhibitor and an anti-infective agent (e.g., a bacteriostatic antibiotic) as described herein and one or more pharmaceutically acceptable additives such as a compatible carrier, binder, filling agent, suspending agent, flavoring agent, sweetening agent, disintegrating agent, dispersing agent, surfactant, lubricant, colorant, diluent, solubilizer, moistening agent, plasticizer, stabilizer, penetration enhancer, wetting agent, anti-foaming agent, antioxidant, preservative, or one or more combination thereof.
  • a compatible carrier such as a compatible carrier, binder, filling agent, suspending agent, flavoring agent, sweetening agent, disintegrating agent, dispersing agent, surfactant, lubricant, colorant, diluent, solubilizer, moistening agent, plasticizer, stabilizer, penetration enhancer, wetting agent, anti-foaming agent, antioxidant, preservative, or one or more combination
  • a film coating is provided around the formulation of the BCL2 family inhibitor and the anti-infective agent.
  • some or all of the particles of these active agents are coated.
  • some or all of the particles of the active agents are microencapsulated.
  • the particles of the active agents are not microencapsulated and are uncoated.
  • Suitable carriers for use in the solid dosage forms described herein include, but are not limited to, acacia, gelatin, colloidal silicon dioxide, calcium glycerophosphate, calcium lactate, maltodextrin, glycerine, magnesium silicate, sodium caseinate, soy lecithin, sodium chloride, tricalcium phosphate, dipotassium phosphate, sodium stearoyl lactylate, carrageenan, monoglyceride, diglyceride, pregelatinized starch, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose acetate stearate, sucrose, microcrystalline cellulose, lactose, mannitol and the like.
  • Suitable filling agents for use in the solid dosage forms described herein include, but are not limited to, lactose, calcium carbonate, calcium phosphate, dibasic calcium phosphate, calcium sulfate, microcrystalline cellulose, cellulose powder, dextrose, dextrates, dextran, starches, pregelatinized starch, hydroxypropylmethycellulose (HPMC), hydroxypropylmethycellulose phthalate, hydroxypropylmethylcellulose acetate stearate (HPMCAS), sucrose, xylitol, lactitol, mannitol, sorbitol, sodium chloride, polyethylene glycol, and the like.
  • disintegrants are often used in the formulation, especially when the dosage forms are compressed with binder. Disintegrants help rupturing the dosage form matrix by swelling or capillary action when moisture is absorbed into the dosage form.
  • Suitable disintegrants for use in the solid dosage forms described herein include, but are not limited to, natural starch such as corn starch or potato starch, a pregelatinized starch such as National 1551 or
  • Amijel , or sodium starch glycolate such as Promogel or Explotab a cellulose such as a wood product, methylcrystalline cellulose, e.g., Avicel ® PH101, Avicel ® PH102, Avicel ® PH105, Elcema ® PI 00, Emcocel ® , Vivacel ® , Ming Tia ® , and Solka-Floc ® , methylcellulose, croscarmellose, or a cross-linked cellulose, such as cross-linked sodium carboxymethylcellulose (Ac-Di-Sol .), cross-linked carboxymethylcellulose, or cross-linked croscarmellose, a cross-linked starch such as sodium starch glycolate, a cross-linked polymer such as crospovidone, a cross-linked polyvinylpyrrolidone, alginate such as alginic acid or a salt of alginic acid such as sodium alginate, a clay such as Veegum
  • Binders impart cohesiveness to solid oral dosage form formulations: for powder filled capsule formulation, they aid in plug formation that can be filled into soft or hard shell capsules and for tablet formulation, they ensure the tablet remaining intact after compression and help assure blend uniformity prior to a compression or fill step.
  • Materials suitable for use as binders in the solid dosage forms described herein include, but are not limited to, carboxymethylcellulose, methylcellulose (e.g., Methocel ® ), hydroxypropylmethylcellulose (e.g.
  • binder levels of 20-70% are used in powder-filled gelatin capsule formulations.
  • Binder usage level in tablet formulations varies whether direct compression, wet granulation, roller compaction, or usage of other excipients such as fillers which itself can act as moderate binder.
  • Formulators skilled in art can determine the binder level for the formulations, but binder usage level of up to 70% in tablet formulations is common.
  • Suitable lubricants or glidants for use in the solid dosage forms described herein include, but are not limited to, stearic acid, calcium hydroxide, talc, corn starch, sodium stearyl fumerate, alkali-metal and alkaline earth metal salts, such as aluminum, calcium, magnesium, zinc, stearic acid, sodium stearates, magnesium stearate, zinc stearate, waxes, Stearowet ., boric acid, sodium benzoate, sodium acetate, sodium chloride, leucine, a polyethylene glycol or a methoxypolyethylene glycol such as Carbowax TM , PEG 4000, PEG 5000, PEG 6000, propylene glycol, sodium oleate, glyceryl behenate, glyceryl palmitostearate, glyceryl benzoate, magnesium or sodium lauryl sulfate, and the like.
  • boric acid sodium benzoate, sodium acetate
  • Suitable diluents for use in the solid dosage forms described herein include, but are not limited to, sugars (including lactose, sucrose, and dextrose), polysaccharides (including dextrates and maltodextrin), polyols (including mannitol, xylitol, and sorbitol), cyclodextrins and the like.
  • Suitable wetting agents for use in the solid dosage forms described herein include, for example, oleic acid, glyceryl monostearate, sorbitan monooleate, sorbitan monolaurate, triethanolamine oleate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monolaurate, quaternary ammonium compounds (e.g., Polyquat 10 ® ), sodium oleate, sodium lauryl sulfate, magnesium stearate, sodium docusate, triacetin, vitamin E TPGS and the like.
  • quaternary ammonium compounds e.g., Polyquat 10 ®
  • sodium oleate sodium lauryl sulfate
  • magnesium stearate sodium docusate
  • triacetin vitamin E TPGS and the like.
  • Suitable surfactants for use in the solid dosage forms described herein include, for example, sodium lauryl sulfate, sorbitan monooleate, polyoxyethylene sorbitan monooleate, polysorbates, polaxomers, bile salts, glyceryl monostearate, copolymers of ethylene oxide and propylene oxide, e.g., Pluronic ® (BASF), and the like.
  • Suitable suspending agents for use in the solid dosage forms described here include, but are not limited to, polyvinylpyrrolidone, e.g., polyvinylpyrrolidone K12, polyvinylpyrrolidone K17, polyvinylpyrrolidone K25, or polyvinylpyrrolidone K30, polyethylene glycol, e.g., the polyethylene glycol can have a molecular weight of about 300 to about 6000, or about 3350 to about 4000, or about 7000 to about 5400, vinyl pyrrolidone/vinyl acetate copolymer (S630), sodium carboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose, polysorbate-80, hydroxyethylcellulose, sodium alginate, gums, such as, e.g., gum tragacanth and gum acacia, guar gum, xanthans, including xanthan gum, sugars, cellulosics, such as,
  • additives used in the solid dosage forms described herein there is considerable overlap between additives used in the solid dosage forms described herein.
  • the above-listed additives should be taken as merely exemplary, and not limiting, of the types of additives that can be included in solid dosage forms described herein.
  • the amounts of such additives can be readily determined by one skilled in the art, according to the particular properties desired.
  • one or more layers of the pharmaceutical formulation are plasticized.
  • a plasticizer is generally a high boiling point solid or liquid. Suitable plasticizers can be added from about 0.01% to about 50% by weight (w/w) of the coating composition.
  • Plasticizers include, but are not limited to, diethyl phthalate, citrate esters, polyethylene glycol, glycerol, acetylated glycerides, triacetin, polypropylene glycol, polyethylene glycol, triethyl citrate, dibutyl sebacate, stearic acid, stearol, stearate, and castor oil.
  • Compressed tablets are solid dosage forms prepared by compacting the bulk blend of the formulations described above.
  • compressed tablets which are designed to dissolve in the mouth will include one or more flavoring agents.
  • the compressed tablets will include a film surrounding the final compressed tablet.
  • the film coating can provide a delayed release of the BCL2 family inhibitor and/or the anti-infective agent.
  • the film coating aids in patient compliance (e.g., Opadry ® coatings or sugar coating). Film coatings including Opadry ® typically range from about 1% to about 3% of the tablet weight.
  • the compressed tablets include one or more excipients.
  • a capsule may be prepared, for example, by placing the bulk blend of the formulation of a BCL2 family inhibitor and anti-infective agent inside of a capsule.
  • the formulations non-aqueous suspensions and solutions
  • the formulations are placed in a soft gelatin capsule.
  • the formulations are placed in standard gelatin capsules or non-gelatin capsules such as capsules comprising HPMC.
  • the formulation is placed in a sprinkle capsule, wherein the capsule may be swallowed whole or the capsule may be opened and the contents sprinkled on food prior to eating.
  • the therapeutic dose is split into multiple (e.g., two, three, or four) capsules.
  • the entire dose of the formulation is delivered in a capsule form.
  • the particles of the BCL2 family inhibitor and anti- infective agent and one or more excipients are dry blended and compressed into a mass, such as a tablet, having a hardness sufficient to provide a pharmaceutical composition that substantially disintegrates within less than about 30 minutes, less than about 35 minutes, less than about 40 minutes, less than about 45 minutes, less than about 50 minutes, less than about 55 minutes, or less than about 60 minutes, after oral administration, thereby releasing the formulation into the gastrointestinal fluid.
  • dosage forms may include microencapsulated formulations. In some embodiments, one or more other compatible materials are present in the microencapsulation material.
  • Exemplary materials include, but are not limited to, pH modifiers, erosion facilitators, anti-foaming agents, antioxidants, flavoring agents, and carrier materials such as binders, suspending agents, disintegration agents, filling agents, surfactants, solubilizers, stabilizers, lubricants, wetting agents, and diluents.
  • carrier materials such as binders, suspending agents, disintegration agents, filling agents, surfactants, solubilizers, stabilizers, lubricants, wetting agents, and diluents.
  • Microencapsulated formulations of a BCL2 family inhibitor and anti-infective agent may be formulated by methods known by one of ordinary skill in the art. Such known methods include, e.g., spray drying processes, spinning disk-solvent processes, hot melt processes, spray chilling methods, fluidized bed, electrostatic deposition, centrifugal extrusion, rotational suspension separation, polymerization at liquid-gas or solid-gas interface, pressure extrusion, or spraying solvent extraction bath. In addition to these, several chemical techniques, e.g., complex coacervation, solvent evaporation, polymer-polymer incompatibility, interfacial polymerization in liquid media, in situ polymerization, in-liquid drying, and desolvation in liquid media could also be used. Furthermore, other methods such as roller compaction, extrusion/spheronization, coacervation, or nanoparticle coating may also be used.
  • the solid dosage formulations of the BCL2 family inhibitor and anti-infective agent are plasticized (coated) with one or more layers.
  • a plasticizer is generally a high boiling point solid or liquid. Suitable plasticizers can be added from about 0.01% to about 50% by weight (w/w) of the coating composition.
  • Plasticizers include, but are not limited to, diethyl phthalate, citrate esters, polyethylene glycol, glycerol, acetylated glycerides, triacetin, polypropylene glycol, polyethylene glycol, triethyl citrate, dibutyl sebacate, stearic acid, stearol, stearate, and castor oil.
  • a powder including the formulations of the BCL2 family inhibitor and anti-infective agent may be formulated to include one or more pharmaceutical excipients and flavors.
  • Such a powder may be prepared, for example, by mixing the formulation and optional pharmaceutical excipients to form a bulk blend composition. Additional embodiments also include a suspending agent and/or a wetting agent. This bulk blend is uniformly subdivided into unit dosage packaging or multi-dosage packaging units.
  • Effervescent powders are also prepared in accordance with the present disclosure.
  • Effervescent salts have been used to disperse medicines in water for oral administration.
  • Effervescent salts are granules or coarse powders containing a medicinal agent in a dry mixture, usually composed of sodium bicarbonate, citric acid and/or tartaric acid.
  • a medicinal agent in a dry mixture, usually composed of sodium bicarbonate, citric acid and/or tartaric acid.
  • the acids and the base react to liberate carbon dioxide gas, thereby causing "effervescence.”
  • effervescent salts include, e.g., the following ingredients: sodium bicarbonate or a mixture of sodium bicarbonate and sodium carbonate, citric acid and/or tartaric acid. Any acid-base combination that results in the liberation of carbon dioxide can be used in place of the combination of sodium bicarbonate and citric and tartaric acids, as long as the ingredients were suitable for pharmaceutical use and result in a pH of about 6.0 or higher.
  • the formulations described herein which include a BCL2 family inhibitor and anti-infective agent, are solid dispersions. Methods of producing such solid dispersions are known in the art and include, but are not limited to, for example, US 4,343,789, 5,340,591, 5,456,923, 5,700,485, 5,723,269, and US 2004/0013734.
  • the formulations described herein are solid solutions. Solid solutions incorporate a substance together with the active agent and other excipients such that heating the mixture results in dissolution of the drug and the resulting composition is then cooled to provide a solid blend which can be further formulated or directly added to a capsule or compressed into a tablet. Methods of producing such solid solutions are known in the art and include, but are not limited to, for example, US 4, 151,273, 5,281,420, and 6,083,518.
  • Controlled release refers to the release of one or more active agents from a dosage form in which they are incorporated according to a desired profile over an extended period of time.
  • Controlled release profiles include, for example, sustained release, prolonged release, pulsatile release, and delayed release profiles.
  • immediate release compositions controlled release compositions allow delivery of an agent to a subject over an extended period of time according to a predetermined profile.
  • Such release rates can provide therapeutically effective levels of agent for an extended period of time and thereby provide a longer period of pharmacologic response while minimizing side effects as compared to conventional rapid release dosage forms. Such longer periods of response provide for many inherent benefits that are not achieved with the corresponding short acting, immediate release preparations.
  • the solid dosage forms described herein can be formulated as enteric coated delayed release oral dosage forms, i.e., as an oral dosage form of a pharmaceutical composition as described herein which utilizes an enteric coating to affect release in the small intestine of the gastrointestinal tract.
  • the enteric coated dosage form may be a compressed or molded or extruded tablet/mold (coated or uncoated) containing granules, powder, pellets, beads or particles of the active ingredient and/or other composition components, which are themselves coated or uncoated.
  • the enteric coated oral dosage form may also be a capsule (coated or uncoated) containing pellets, beads or granules of the solid carrier or the composition, which are themselves coated or uncoated.
  • delayed release refers to the delivery so that the release can be accomplished at some generally predictable location in the intestinal tract more distal to that which would have been accomplished if there had been no delayed release alterations.
  • the method for delay of release is coating. Any coatings should be applied to a sufficient thickness such that the entire coating does not dissolve in the gastrointestinal fluids at pH below about 5, but does dissolve at pH about 5 and above. It is expected that any anionic polymer exhibiting a pH-dependent solubility profile can be used as an enteric coating in the methods and compositions described herein to achieve delivery to the lower gastrointestinal tract.
  • the polymers described herein are anionic carboxylic polymers.
  • the coating can, and usually does, contain a plasticizer and possibly other coating excipients such as colorants, talc, and/or magnesium stearate, which are well known in the art.
  • Suitable plasticizers include triethyl citrate (Citroflex 2), triacetin (glyceryl triacetate), acetyl triethyl citrate (Citroflec A2), Carbowax 400 (polyethylene glycol 400), diethyl phthalate, tributyl citrate, acetylated monoglycerides, glycerol, fatty acid esters, propylene glycol, and dibutyl phthalate.
  • anionic carboxylic acrylic polymers usually will contain 10-25% by weight of a plasticizer, especially dibutyl phthalate, polyethylene glycol, triethyl citrate and triacetin.
  • a plasticizer especially dibutyl phthalate, polyethylene glycol, triethyl citrate and triacetin.
  • Conventional coating techniques such as spray or pan coating are employed to apply coatings. The coating thickness must be sufficient to ensure that the oral dosage form remains intact until the desired site of topical delivery in the intestinal tract is reached.
  • Colorants, detackifiers, surfactants, antifoaming agents, lubricants may be added to the coatings besides plasticizers to solubilize or disperse the coating material, and to improve coating performance and the coated product.
  • the BCL2 family inhibitor plus anti-infective agent formulations described herein are delivered using a pulsatile dosage form.
  • a pulsatile dosage form is capable of providing one or more immediate release pulses at predetermined time points after a controlled lag time or at specific sites.
  • Pulsatile dosage forms may be administered using a variety of pulsatile formulations known in the art. For example, such formulations include, but are not limited to, those described in US 5,011,692, 5,017,381, 5,229, 135, and 5,840,329.
  • the controlled release dosage form is pulsatile release solid oral dosage form including at least two groups of particles, (i.e. multiparticulate) each containing a formulation described herein.
  • the first group of particles provides a substantially immediate dose of the BCL2 family inhibitor and anti-infective agent upon ingestion.
  • the first group of particles can be either uncoated or include a coating and/or sealant.
  • the second group of particles includes coated particles, which includes from about 2% to about 75%, from about 2.5% to about 70%, or from about 40% to about 70%), by weight of the total dose of the active agents in the formulation, in admixture with one or more binders.
  • the coating includes a pharmaceutically acceptable ingredient in an amount sufficient to provide a delay of from about 2 hours to about 7 hours following ingestion before release of the second dose.
  • Suitable coatings include one or more differentially degradable coatings such as, by way of example only, pH sensitive coatings (enteric coatings) such as acrylic resins either alone or blended with cellulose derivatives, e.g., ethylcellulose, or non-enteric coatings having variable thickness to provide differential release of the formulation.
  • controlled release systems include, e.g., polymer-based systems, such as polylactic and polyglycolic acid, plyanhydrides and polycaprolactone; porous matrices, nonpolymer-based systems that are lipids, including sterols, such as cholesterol, cholesterol esters and fatty acids, or neutral fats, such as mono-, di- and triglycerides; hydrogel release systems; silastic systems; peptide-based systems; wax coatings, bioerodible dosage forms, compressed tablets using conventional binders and the like. See, e.g., Liberman et al.
  • pharmaceutical formulations include particles of a BCL2 family inhibitor and anti-infective agent, and at least one dispersing agent or suspending agent for oral administration to a subject.
  • the formulations may be a powder and/or granules for suspension, and upon admixture with water, a substantially uniform suspension is obtained.
  • Liquid formulation dosage forms for oral administration can be aqueous suspensions selected from the group including, but not limited to, pharmaceutically acceptable aqueous oral dispersions, emulsions, solutions, elixirs, gels, and syrups. See, e.g., Singh et al. (2002).
  • aqueous suspensions and dispersions described herein can remain in a homogenous state, as defined in The USP Pharmacists' Pharmacopeia (2005 edition, chapter 905), for at least 4 hours.
  • the homogeneity should be determined by a sampling method consistent with regard to determining homogeneity of the entire composition.
  • an aqueous suspension can be re-suspended into a homogenous suspension by physical agitation lasting less than 1 minute.
  • an aqueous suspension can be re-suspended into a homogenous suspension by physical agitation lasting less than 45 seconds.
  • an aqueous suspension can be re-suspended into a homogenous suspension by physical agitation lasting less than 30 seconds. In still another embodiment, no agitation is necessary to maintain a homogeneous aqueous dispersion.
  • the liquid formulations can also include inert diluents commonly used in the art, such as water or other solvents, solubilizing agents, and emulsifiers.
  • emulsifiers are ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol, dimethylformamide, sodium lauryl sulfate, sodium doccusate, cholesterol, cholesterol esters, taurocholic acid, phosphotidylcholine, oils, such as cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil, and sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols, fatty acid esters of sorbitan, or mixtures of these substances, and the like.
  • Intranasal formulations are known in the art and are described in, for example, US 4,476, 116, 5, 116,817 and 6,391,452.
  • Formulations prepared according to these and other techniques well-known in the art are prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, fluorocarbons, and/or other solubilizing or dispersing agents known in the art. See, for example, Ansel, H. C. et al. (1995).
  • these compositions and formulations are prepared with suitable nontoxic pharmaceutically acceptable ingredients.
  • nasal dosage forms generally contain large amounts of water in addition to the active ingredient. Minor amounts of other ingredients such as pH adjusters, emulsifiers or dispersing agents, preservatives, surfactants, gelling agents, or buffering and other stabilizing and solubilizing agents may also be present.
  • the nasal dosage form should be isotonic with nasal secretions.
  • formulations of a BCL2 family inhibitor and an anti-infective agent may be in the form of an aerosol, a mist, or a powder.
  • Pharmaceutical compositions described herein are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebuliser, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • a suitable propellant e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • the dosage unit may be determined by providing a valve to deliver a metered amount.
  • Capsules and cartridges of, such as, by way of example only, gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound described herein and a suitable powder base such as lactose or starch.
  • a powder mix of the compound described herein and a suitable powder base such as lactose or starch.
  • buccal formulations are known in the art and are described in, for example, US 4,229,447, 4,596,795, 4,755,386, and 5,739,136.
  • the buccal dosage forms described herein can further include a bioerodible (hydrolysable) polymeric carrier that also serves to adhere the dosage form to the buccal mucosa.
  • the buccal dosage form is fabricated so as to erode gradually over a predetermined time period, wherein the delivery of the active agents is provided essentially throughout.
  • Buccal drug delivery avoids the disadvantages encountered with oral drug administration, e.g., slow absorption, degradation of active agents by fluids present in the gastrointestinal tract and/or first- pass inactivation in the liver.
  • the polymeric carrier comprises hydrophilic (water-soluble and water- swellable) polymers that adhere to the wet surface of the buccal mucosa.
  • hydrophilic water-soluble and water- swellable
  • polymeric carriers useful herein include acrylic acid polymers and co, e.g., those known as "carbomers”.
  • compositions may take the form of tablets, lozenges, or gels formulated in a conventional manner.
  • Transdermal dosage formulations of a BCL2 family inhibitor and anti- infective agent described herein may incorporate certain pharmaceutically acceptable excipients which are conventional in the art.
  • the transdermal formulations described herein include at least three components: (1) a formulation of a BCL2 family inhibitor and an anti-infective agent; (2) a penetration enhancer; and (3) an aqueous adjuvant.
  • transdermal formulations can include additional components such as, but not limited to, gelling agents, creams and ointment bases, and the like.
  • the transdermal formulation can further include a woven or non- woven backing material to enhance absorption and prevent the removal of the transdermal formulation from the skin.
  • the transdermal formulations described herein can maintain a saturated or supersaturated state to promote diffusion into the skin.
  • Formulations suitable for transdermal administration of compounds described herein may employ transdermal delivery devices and transdermal delivery patches and can be lipophilic emulsions or buffered, aqueous solutions, dissolved and/or dispersed in a polymer or an adhesive. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents. Still further, transdermal delivery of the compounds described herein can be accomplished by means of iontophoretic patches and the like. Additionally, transdermal patches can provide controlled delivery of active agents. The rate of absorption can be slowed by using rate-controlling membranes or by trapping the compound within a polymer matrix or gel. Conversely, absorption enhancers can be used to increase absorption.
  • transdermal devices are in the form of a bandage comprising a backing member, a reservoir containing the compound optionally with carriers, optionally a rate controlling barrier to deliver the compound to the skin of the host at a controlled and predetermined rate over a prolonged period of time, and means to secure the device to the skin.
  • Transdermal formulations may be administered using a variety of devices which have been described in the art.
  • such devices include, but are not limited to, US 3,598, 122, 3,598, 123, 3,710,795, 3,731,683, 3,742,951, 3,814,097, 3,921,636, 3,972,995, 3,993,072, 3,993,073, 3,996,934, 4,031,894, 4,060,084, 4,069,307, 4,077,407, 4,201,211, 4,230, 105, 4,292,299, 4,292,303, 5,336, 168, 5,665,378, 5,837,280, 5,869,090, 6,923,983, 6,929,801 and 6,946, 144.
  • Formulations suitable for intramuscular, subcutaneous, or intravenous injection may include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions.
  • suitable aqueous and nonaqueous carriers, diluents, solvents, or vehicles including water, ethanol, polyols (propyleneglycol, polyethylene-glycol, glycerol, cremophor and the like), suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate.
  • Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.
  • Formulations suitable for subcutaneous injection may also contain additives such as preserving, wetting, emulsifying, and dispensing agents. Prevention of the growth of microorganisms can be ensured by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, such as aluminum monostearate and gelatin.
  • compounds described herein may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiological saline buffer.
  • physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiological saline buffer.
  • penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
  • appropriate formulations may include aqueous or nonaqueous solutions, preferably with physiologically compatible buffers or excipients. Such excipients are generally known in the art.
  • Parenteral injections may involve bolus injection or continuous infusion.
  • Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative.
  • the pharmaceutical composition described herein may be in a form suitable for parenteral injection as a sterile suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
  • Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions.
  • Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes.
  • Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran.
  • the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.
  • the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen- free water, before use.
  • compositions described herein may be in unit dosage forms suitable for single administration of precise dosages.
  • unit dosage form the formulation is divided into unit doses containing appropriate quantities of one or more compound.
  • the unit dosage may be in the form of a package containing discrete quantities of the formulation.
  • Non-limiting examples are packaged tablets or capsules, and powders in vials or ampoules.
  • Aqueous suspension compositions can be packaged in single-dose non-reclosable containers.
  • multiple-dose reclosable containers can be used, in which case it is typical to include a preservative in the composition.
  • formulations for parenteral injection may be presented in unit dosage form, which include, but are not limited to ampoules, or in multi-dose containers, with an added preservative.
  • kits for use in the therapeutic applications described herein include at least a BCL2 family inhibitor and instructions for use of the BCL2 family inhibitor for treatment of an infection according to any of the methods described herein.
  • the BCL2 family inhibitor is a BH3 mimetic agent.
  • the BH3 mimetic agent inhibits BCL- XL.
  • the BH3 mimetic agent inhibits BCL-XL preferentially relative to other BCL2 family members.
  • a kit containing the BCL2 family inhibitor also includes an anti-infective agent.
  • the kit includes a BCL2 family inhibitor and bacteriostatic or bactericidal agent (e.g., an antibiotic).
  • the kit includes a BCL2 family inhibitor and an anti-viral agent. In further embodiments the kit includes a BCL2 family inhibitor and antifungal agent. In yet other embodiments the kit includes a BCL2 family inhibitor and an antiprotozoal agent.
  • kits include a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) including one of the separate elements to be used in a method described herein.
  • Suitable containers include, for example, bottles, vials, syringes, and test tubes.
  • the containers can be formed from a variety of materials such as glass or plastic.
  • a kit will typically may include one or more additional containers, each with one or more of various materials (such as reagents, optionally in concentrated form, and/or devices) desirable from a commercial and user standpoint for use of a compound described herein.
  • materials include, but not limited to, buffers, diluents, filters, needles, syringes; carrier, package, container, vial and/or tube labels listing contents and/or instructions for use, and package inserts with instructions for use.
  • a set of instructions for performing at least one of the treatment methods described herein is typically also included.
  • a label can be on or associated with the container.
  • a label can be on a container when letters, numbers or other characters forming the label are attached, molded or etched into the container itself; a label can be associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert.
  • a label can be used to indicate that the contents are to be used for treatment of an infection (e.g., a bacterial infection).
  • the label can also indicate directions for use of the contents, such as in the methods described herein.
  • L. pneumophila 130b sero-group 1 is a spectinomycin- resistant clinical isolate from the Wadsworth Veterans Administration Hospital, Los Angeles, CA (Edelstein et al, 1986).
  • the avirulent AdotA and the flagellin-deficient AflaA strains are deletion mutants of L. pneumophila 130b (Harding et al, 2013).
  • GFP was expressed constitutively using the plasmid pMMB207C.
  • longbeachae NSW- 150 a serogroup 1 clinical isolate from Australia, were grown from -80 °C frozen stocks on buffered charcoal-yeast extract (BCYE) agar at 37 °C for 48 h before each infection.
  • BCYE buffered charcoal-yeast extract
  • chloramphenicol was used at 6 ⁇ g.mL "1 .
  • OD 6 oo optical density at 600 nm
  • Murine bone marrow-derived macrophages were obtained from femora and tibiae of female 6-8 week-old C57BL/6 mice, or from mice of the indicated genotypes. Macrophages were cultured in RPMI 1640 medium supplemented with 15% fetal bovine serum (Serana), 20% L-cell-conditioned medium (containing macrophage colony-stimulating factor), and 100 U/mL of penicillin-streptomycin (Sigma) in bacteriological dishes for 12 days, at 37 °C + 5% C0 2 . Medium was replaced after one week.
  • BMDMs were gently scraped from plates using a cell scraper (BD Falcon) and washed 3 times in PBS, before seeding into tissue culture-treated plates.
  • Immortalised C57BL/6 mouse derived macrophages (Bauernfeind et al, 2009) were cultured in RPMI 1640 supplemented with 10 % fetal bovine serum, at 37 °C and 5 % C0 2 . Live Cell Imaging to Determine Macrophage Viability
  • Cells were seeded at a density of 5 ⁇ 10 5 cells/mL in 96-well tissue culture plates and cell death was determined essentially as described in (Croker et al, 201 1). Cells were stained with 1 ⁇ Cell Tracker Green (CTG) (Invitrogen) for 20 min in serum- free RPMI 1640. Medium was then replaced with RPMI 1640 supplemented with 15 % FBS and 10 % L-cell-conditioned medium. Cells were stained with 0.6 ⁇ Draq7 (Abeam) or 1 ⁇ g/mL propidium iodide (Invitrogen).
  • CCG Cell Tracker Green
  • Time-lapse images were acquired with bright-field, GFP, TxRed, and Y5 filters every hour for up to 72 h using a 10 ⁇ /0.8- ⁇ objective.
  • images were analyzed in ImageJ and in MetaMorph® (Molecular Devices) using a custom-made journal suite incorporating the count nuclei function to segment and count the number of CTG and Draq7 positive cells (adapted from Croker et al, 2011).
  • the data of percentage Draq7-positive cells were analyzed in Excel and GraphPad Prism.
  • macrophages were seeded at a density of 2.5 ⁇ 10 5 cells/mL into 12-well tissue culture plates and infected with Legionella strains expressing GFP at an MOI of 50. After 2 h, cells were washed 3 ⁇ in PBS, medium replaced, and cells were then treated with BH3 mimetics [500 nM] and Q-VD-OPh [20 ⁇ ]. At the indicated time points, cells were removed from the plates and stained with 1 ⁇ g/mL propidium iodide (PI) (Invitrogen) before GFP and PI fluorescence were determined by flow cytometric analysis (BD FACSCalibur TM ).
  • PI propidium iodide
  • Cells were gated by forward and side scatter, and channels Fl and F3 were used to detect GFP and PI fluorescence, respectively. 10.000 events/sample were counted. Weasel software (WEHI) was used for the analysis. Alternatively, cells were lysed in 0.05 % digitonin for 5 min at room temperature and serial dilutions of the cell lysates and the corresponding culture media were plated on BCYE agar plates, and bacterial colonies counted after 72 h at 37 °C.
  • WEHI Weasel software
  • mice 6-8 week-old male or female C57BL/6 and A/J mice (or mice of the indicated genotypes), in groups of five or more, were anesthetized by 4 % isofluorane inhalation and infected intra-nasally with either 2.5 x 10 6 L. pneumophila, or 1 x 10 5 L. longbeachae, in 50 ⁇ _, of sterile PBS.
  • mice were injected intra-peritoneally (i.p.) with 50 mg per kg body weight of ABT-263 (Selleckchem) in 50 ⁇ . DMSO, or with DMSO (vehicle, control) only.
  • mice were weighed daily and euthanized after greater than 15% weight loss, according to the animal ethics guidelines.
  • Cre-mediated Bcl-xl deletion was induced by 3 doses of 200mg/kg tamoxifen (Sigma, T5648) dissolved in peanut oil/10% ethanol at 80 mg/ml administered on three separate days by oral gavage using bulb-tipped feeding needles.
  • membranes were probed with secondary goat anti-rabbit IgG (Life Technologies) and goat anti-mouse IgG (Life Technologies) antibodies conjugated to HRP (1 :20,000 dilution in T-BST + 5 % skim milk).
  • HRP 1 :20,000 dilution in T-BST + 5 % skim milk.
  • Membranes were developed with the luminol-based enhanced chemiluminescence (ECL) and exposed to film (Kodak). Scanned images were processed in Adobe Photoshop.
  • Macrophages were seeded onto glass coverslips in 24-well plates and infected with Legionella strains expressing GFP at an MOI of 10.
  • BH3 mimetics [500 nM] were added to appropriate wells, 2 h after infection.
  • cells were fixed with 4 % PFA for 15 min, washed three times with PBS and treated with 50 mM H 4 C1 for 10 min.
  • Cells were then permeabilized in 0.1 % Triton-X 100 in PBS for 5 min on ice and blocked in PBS + 1 % BSA overnight at 4 °C, before incubation with anti-cleaved caspase-3 antibody (CST #9964) [1 :400] for 30 min.
  • CST #9964 anti-cleaved caspase-3 antibody
  • Example 2 The BH3 Mimetic Compound ABT-737 Inhibits Induces Apoptosis of Legionella-Infected Bone Marrow-Derived Macrophages
  • the inventors used the flagellin-deficient AflaA mutant, as AflaA -infected BMDMs remain viable for up to 20 h post-infection, allowing intracellular replication (Figure la). This mimics wild- type Legionella infection in permissive human macrophages (Miao et al, 2010; Ren et al, 2006; Molofsky et al, 2006; Zamboni et al, 2006; and Amer et al, 2006). Subsequent macrophage death occurred in -80 % of BMDMs by 60 h post-infection (Figure la), reflecting cycles of bacterial release and re-infection, and continual bacterial replication at a low initial multiplicity of infection.
  • the BH3 mimetic compound, ABT-737 antagonizes pro-survival BCL-2 family members to cause cell death in some cells (Lessene et al, 2008).
  • AflaA L. pneumophi la-infected BMDMs were treated with ABT-737 the majority of macrophages ( ⁇ 65 %) were protected from the extensive cell death that occurred in untreated cells 60 h post infection ( Figure la).
  • ABT-737 treatment protected BMDMs from extensive cell death after infection with wild type L. pneumophila ( Figure 5b). In both cases ABT-737 treatment resulted in 30-35% BMDM death ( Figure la and Figure 5b).
  • ABT-737 might induce selective killing of the initial Legionella- arborm ' g macrophages (-30-40% of cells) shortly after bacterial invasion. Consistent with this idea, at higher infection rates, ABT-737 treatment resulted in the killing of -80 % of BMDMs in culture, and this occurred faster than in infected macrophages that had not been exposed to ABT-737 (Figure lc). Moreover, live-cell imaging of immortalized C57BL/6 (iBl/6) macrophages after infection with GFP-expressing AflaA L.
  • ABT-737 treatment reduces intracellular L. pneumophila loads
  • CFUs colony forming units
  • ABT-737 targets BCL-2, BCL-W and BCL-XL ( Figure 2a).
  • BCL-2 member protects Legionella-m ' iected macrophages from death
  • the inventors utilized BH3-mimetics that specifically bind to BCL-2 or BCL-XL.
  • the BCL-2-specific antagonist, ABT-199 failed to clear GFP-expressing AflaA L. pneumophila in iBL/6 and BMDM host cells ( Figure 2a, b and c).
  • A-l 155463 In contrast to ABT-199, the BCL-XL-specific inhibitor A-l 155463, which does not antagonize BCL-2 or BCL-W significantly (Tao et al, 2014), mimicked the effects of ABT-737. Specifically, like ABT-737, A-l 155463 treatment caused the loss of GFP signal in iBl/6 macrophages after infection with GFP-expressing AflaA L. pneumophila, and resulted in a > 100-fold CFU reduction in BMDMs 48 h post infection ( Figure 2a and c). Consequently, A-l 155463 protected macrophages from extensive Legionella- mediated killing ( Figure 2a and b). Remarkably, A-l 155463 was effective in limiting AflaA Legionella burdens even at low nanomolar concentrations, whereas ABT-199 was inefficient at doses lower than 5 ⁇ ( Figure 2d).
  • BCL-XL-deficient BMDMs To genetically confirm a role for BCL-XL in promoting Legionella infection, the inventors generated BCL-XL-deficient BMDMs. Constitutive loss of BCL-XL causes embryonic lethality around E14 due to excessive apoptosis of erythroid and neuronal progenitors (Motoyama et al, 1995).
  • Bcl-x flox/flox ;ER-Cre mice, which contain two floxed Bcl-x alleles (Wagner et al, 2000) and express the Cre-recombinase estrogen receptor fusion protein (Rosa26-Cre-ERT2), with 4-hydroxytamoxifen (4-HT) to induce deletion of BCL-XL (referred to as Bcl-x ' ' ).
  • Bcl-x ' ' 4-hydroxytamoxifen
  • BCL-XL inhibits the mitochondrial apoptotic pathway that culminates in the activation of the apoptotic executioner caspase-3 (Strasser et al, 2011). Consistent with this, AflaA, but not AdotA, L. pneumophi /a-infected Bcl-x ' ' BMDMs showed increased caspase-3 activation compared to wild type BMDMs, as measured by immunoblotting or staining of the active caspase-3 pl7/pl9 fragment (Figs. 3a and c). Increased caspase-3 activation was also detected in AflaA L. pneumophi la-infected wild type BMDMs treated with A-l 155463 ( Figure 3b). In contrast, comparatively less caspase- 3 activation was detected in AflaA -infected wild type macrophages that had been treated with vehicle control ( Figure 3b).
  • mice were treated >10-week-old Bcl- ⁇ ox, ⁇ m ; ER-Cre mice with tamoxifen to generate BLC-XL deficient mice. These mice contained normal numbers of bone marrow progenitor cells, which generated BMDMs that lacked detectable levels of BCL-XL protein ( Figure 4a), and showed reduced bacterial loads 48 h after AflaA L. pneumophila infection ( Figure 7a). Next, tamoxifen-treated or untreated Bcl- ⁇ ox, fl m ;ER-Cre mice were intranasally infected with AflaA L. pneumophila, and the bacterial burden in the lungs determined at 48 h post-infection.
  • mice which are susceptible to wild type L. pneumophila.
  • mice were treated with a single clinically relevant (50 mg/kg body weight) dose of the orally available analogue of ABT-737 (ABT-263). This resulted in a significant reduction (p ⁇ 0.01) in the numbers of bacteria in the lungs at 48 h post-infection ( Figure 4c).
  • ABT-263 the orally available analogue of ABT-737
  • pneumophila causes a self-limiting infection even in susceptible A/J mice (Molofsky et al, 2006), the inventors also tested whether targeting BCL-XL would affect Legionella longbeachae, which grows in mouse lungs unrestrained (Gobin et al, 2009; and Asare et al, 2007).
  • L. longbeachae causes lethal pneumonia by employing a different set of T4SS effectors (Cazalet et al, 2010).
  • ABT-737 and A-l 155463 both prevented increased intracellular loads of L. longbeachae in BMDMs ( Figure 8a).
  • Administration of ABT-263 effectively controlled the burden of L.
  • Example 4 A BCL-XL-Selective BH3 Mimetic Compound, Reduces Bacterial Load In Vivo and Extends Survival
  • mice Eight week-old female C57BL/6 mice were infected intranasally with 10 5 WT Legionella longbeachae bacteria and received either a single dose of 50 mg/kgof a BCL-XL-selective BH3 mimetic compound (Formula XIII), i.p., or a DMSO (control) injection. Mice were culled 72 h post-infections, lungs extracted, and serial dilutions plated onto BCYE plates, and incubated for three days at 37 C prior to colony counting. Similar to ABT-263, BCL-XL inhibition caused a 100 fold reduction in bacterial counts.
  • Example 5 A BCL-XL-Selective BH3 Mimetic Compound.
  • A-1331852 (Formula XIII) is Effective in Treament of a Model of Latent Mycobacterium tuberculosis infection
  • mice were infected using an inhalation exposure system with a low dose of M.tb (70-100 CFUs). At three weeks, they were commenced on isoniazid (O. lmg/L) and rifampicin (0.075mg/L) ad libitum for a total of 12 weeks. Mice were rested for two weeks, prior to treatment with A-1331852 at 25 mg/kg daily via gavage for 10 days. After a week of rest, mice were then injected intraperitoneally with dexamethasone 120 ⁇ g for 6 doses. Mice were then sacrificed after one week for CFU calculations.
  • isoniazid O. lmg/L
  • rifampicin 0.075mg/L

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Virology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oncology (AREA)
  • Communicable Diseases (AREA)
  • Molecular Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Pulmonology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • AIDS & HIV (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne des méthodes de traitement d'une infection chez le patient le nécessitant. En particulier, la présente invention concerne le traitement d'une infection par administration d'une quantité thérapeutiquement efficace d'un inhibiteur des protéines de la famille BCL2 (par exemple, un BH3-mimétique), seul ou en association avec un agent anti-infectieux. L'invention concerne également des méthodes et des compositions, y compris des compositions pharmaceutiques, contenant à la fois un inhibiteur des protéines de la famille BCL2 et un agent anti-infectieux. L'invention concerne également des kits contenant à la fois un inhibiteur des protéines de la famille BCL2 et un agent anti-infectieux, soit fournis séparément soit associés au sein d'une formulation unique.
PCT/AU2016/050105 2015-02-18 2016-02-17 Méthodes de traitement de maladies infectieuses WO2016131100A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
AU2015900554A AU2015900554A0 (en) 2015-02-18 BCL2 Family Inhibition for the Treatment of Infectious Disease
AU2015900554 2015-02-18

Publications (1)

Publication Number Publication Date
WO2016131100A1 true WO2016131100A1 (fr) 2016-08-25

Family

ID=56691956

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/AU2016/050105 WO2016131100A1 (fr) 2015-02-18 2016-02-17 Méthodes de traitement de maladies infectieuses

Country Status (1)

Country Link
WO (1) WO2016131100A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108261414A (zh) * 2018-01-11 2018-07-10 广西师范大学 一种治疗肺癌的药物组合物
WO2019067977A1 (fr) * 2017-09-29 2019-04-04 The George Washington University Utilisation d'agents senolytiques pour éliminer des réservoirs de vih persistants
CN111801320A (zh) * 2018-01-10 2020-10-20 里科瑞尔姆Ip控股有限责任公司 苯甲酰胺化合物
FR3108503A1 (fr) * 2020-03-24 2021-10-01 Hermine Lanniel Composition pharmaceutique active contre une infection virale du type coronavirus
WO2024012557A1 (fr) * 2022-07-15 2024-01-18 Berrybio (Hong Kong) Limited Agents de dégradation de protéines anti-apoptotiques de la famille bcl-2, compositions pharmaceutiques et applications thérapeutiques

Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000004901A1 (fr) * 1998-07-21 2000-02-03 Thomas Jefferson University Petites molecules inhibitrices de proteines bcl-2
WO2005049594A1 (fr) * 2003-11-13 2005-06-02 Abbott Laboratories Promoteurs de l'apoptose contenant n-acylsulfonamide
WO2007040650A2 (fr) * 2005-05-12 2007-04-12 Abbott Laboratories Promoteurs d'apoptose
WO2010068684A2 (fr) * 2008-12-09 2010-06-17 Dana Farber Cancer Institute, Inc. Procédés et compositions pour la modulation spécifique de mcl-1
WO2010080503A1 (fr) * 2008-12-19 2010-07-15 Genentech, Inc. Composés hétérocycliques et leurs procédés d'utilisation
WO2010080478A1 (fr) * 2008-12-19 2010-07-15 Genentech, Inc. Composés et méthodes d'utilisation
WO2011094708A2 (fr) * 2010-01-29 2011-08-04 Dana-Farber Cancer Institute, Inc Petites molécules pour la modulation de mcl-1 et procédés de modulation de la mort cellulaire, la division cellulaire, la différenciation cellulaire et procédés de traitement de troubles
WO2013055895A1 (fr) * 2011-10-14 2013-04-18 Abbvie Inc. Agents d'induction de l'apoptose pour traitement du cancer et de maladies immunitaires et auto-immunes
WO2013055897A1 (fr) * 2011-10-14 2013-04-18 Abbvie Inc. Dérivés 8-carbamoyl-2-(2,3-di-substitué pyrid-6-yl)-1,2,3,4-tétrahydroisoquinoléine en tant qu'agents induisant une apoptose pour le traitement du cancer et de maladies immunes et auto-immunes
WO2014028381A1 (fr) * 2012-08-13 2014-02-20 Abbvie Inc. Agents induisant l'apoptose
WO2015126912A1 (fr) * 2014-02-18 2015-08-27 H. Lee Moffitt Cancer Center And Research Institute, Inc. Dérivés marinopyrrole et procédés de production et d'utilisation de ces composés

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000004901A1 (fr) * 1998-07-21 2000-02-03 Thomas Jefferson University Petites molecules inhibitrices de proteines bcl-2
WO2005049594A1 (fr) * 2003-11-13 2005-06-02 Abbott Laboratories Promoteurs de l'apoptose contenant n-acylsulfonamide
WO2007040650A2 (fr) * 2005-05-12 2007-04-12 Abbott Laboratories Promoteurs d'apoptose
WO2010068684A2 (fr) * 2008-12-09 2010-06-17 Dana Farber Cancer Institute, Inc. Procédés et compositions pour la modulation spécifique de mcl-1
WO2010080503A1 (fr) * 2008-12-19 2010-07-15 Genentech, Inc. Composés hétérocycliques et leurs procédés d'utilisation
WO2010080478A1 (fr) * 2008-12-19 2010-07-15 Genentech, Inc. Composés et méthodes d'utilisation
WO2011094708A2 (fr) * 2010-01-29 2011-08-04 Dana-Farber Cancer Institute, Inc Petites molécules pour la modulation de mcl-1 et procédés de modulation de la mort cellulaire, la division cellulaire, la différenciation cellulaire et procédés de traitement de troubles
WO2013055895A1 (fr) * 2011-10-14 2013-04-18 Abbvie Inc. Agents d'induction de l'apoptose pour traitement du cancer et de maladies immunitaires et auto-immunes
WO2013055897A1 (fr) * 2011-10-14 2013-04-18 Abbvie Inc. Dérivés 8-carbamoyl-2-(2,3-di-substitué pyrid-6-yl)-1,2,3,4-tétrahydroisoquinoléine en tant qu'agents induisant une apoptose pour le traitement du cancer et de maladies immunes et auto-immunes
WO2014028381A1 (fr) * 2012-08-13 2014-02-20 Abbvie Inc. Agents induisant l'apoptose
WO2015126912A1 (fr) * 2014-02-18 2015-08-27 H. Lee Moffitt Cancer Center And Research Institute, Inc. Dérivés marinopyrrole et procédés de production et d'utilisation de ces composés

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
KOZIEL, J. ET AL.: "The role of Mcl-1 in S. aureus-induced cytoprotection of infected macrophages", MEDIATORS OF INFLAMMATION, 2013, pages 1 - 12 *
LABELLE, J. L. ET AL.: "A stapled BIM peptide overcomes apoptotic resistance in hematologic cancers", THE JOURNAL OF CLINICAL INVESTIGATION, vol. 122, no. 6, 2012, pages 2018 - 2031, XP055189510, DOI: doi:10.1172/JCI46231 *
LUCAS, C.D. ET AL.: "Downregulation of Mcl-1 has anti-inflammatory pro-resolution effects and enhances bacterial clearance from the lung", MUCOSAL IMMUNOLOGY, vol. 7, no. 4, 2014, pages 857 - 868 *
TSUJIMOTO, Y.: "Role of Bcl-2 family proteins in apoptosis: apoptosomes or mitochondria?", GENES TO CELLS, vol. 3, 1998, pages 697 - 707 *
WALENSKY, L. D. ET AL.: "Activation of apoptosis in vivo by a hydrocarbon-stapled BH3 helix", SCIENCE, vol. 305, no. 5689, 2004, pages 1466 - 1470, XP002555229, DOI: doi:10.1126/science.1099191 *

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019067977A1 (fr) * 2017-09-29 2019-04-04 The George Washington University Utilisation d'agents senolytiques pour éliminer des réservoirs de vih persistants
EP3740487A4 (fr) * 2018-01-10 2021-11-10 Recurium IP Holdings, LLC Composés benzamide
JP2021510376A (ja) * 2018-01-10 2021-04-22 リキュリウム アイピー ホールディングス リミテッド ライアビリティー カンパニー ベンズアミド化合物
EP3737681A4 (fr) * 2018-01-10 2022-01-12 Recurium IP Holdings, LLC Composés benzamide
CN111801327A (zh) * 2018-01-10 2020-10-20 里科瑞尔姆Ip控股有限责任公司 苯甲酰胺化合物
US11318134B2 (en) 2018-01-10 2022-05-03 Recurium Ip Holdings, Llc Benzamide compounds
EP3737672A4 (fr) * 2018-01-10 2021-09-15 Recurium IP Holdings, LLC Composés benzamide
CN111801327B (zh) * 2018-01-10 2024-02-09 里科瑞尔姆Ip控股有限责任公司 苯甲酰胺化合物
US11344546B2 (en) 2018-01-10 2022-05-31 Recurium IP Holding, LLC Benzamide compounds
CN111801320A (zh) * 2018-01-10 2020-10-20 里科瑞尔姆Ip控股有限责任公司 苯甲酰胺化合物
US11813260B1 (en) 2018-01-10 2023-11-14 Recurium Ip Holdings, Llc Benzamide compounds
US11813259B2 (en) 2018-01-10 2023-11-14 Recurium Ip Holdings, Llc Benzamide compounds
US11590126B2 (en) 2018-01-10 2023-02-28 Recurium Ip Holdings, Llc Benzamide compounds
JP7355741B2 (ja) 2018-01-10 2023-10-03 リキュリウム アイピー ホールディングス リミテッド ライアビリティー カンパニー ベンズアミド化合物
CN108261414A (zh) * 2018-01-11 2018-07-10 广西师范大学 一种治疗肺癌的药物组合物
CN108261414B (zh) * 2018-01-11 2019-09-06 广西师范大学 一种治疗肺癌的药物组合物
FR3108503A1 (fr) * 2020-03-24 2021-10-01 Hermine Lanniel Composition pharmaceutique active contre une infection virale du type coronavirus
WO2024012557A1 (fr) * 2022-07-15 2024-01-18 Berrybio (Hong Kong) Limited Agents de dégradation de protéines anti-apoptotiques de la famille bcl-2, compositions pharmaceutiques et applications thérapeutiques

Similar Documents

Publication Publication Date Title
US10617735B2 (en) Methods and compositions for preserving retinal ganglion cells
AU2012321110B2 (en) Combination treatment
WO2016131100A1 (fr) Méthodes de traitement de maladies infectieuses
JP7047148B2 (ja) 骨髄腫を治療するためのセルデュラチニブ（ｃｅｒｄｕｌａｔｉｎｉｂ）
DK2337575T3 (en) A method of treatment with single doses of oritavancin
KR20080018874A (ko) 인간 질병과 관련된 돌연변이 단백질의 분해 촉진을 위한물질 및 방법
Sharma et al. Multidrug resistance crisis during COVID-19 pandemic: role of anti-microbial peptides as next-generation therapeutics
RU2592983C2 (ru) Комбинированная терапия, включающая вемурафениб и интерферон, для применения в лечении рака
US11273170B2 (en) Methods of treating cancer
JP6837700B2 (ja) ジピベフリンの使用方法
JP5550906B2 (ja) 嚢胞性疾患の治療のためのプリン誘導体
BR112021006132A2 (pt) compostos bifenil sulfonamida para o tratamento de doenças de colágeno tipo iv
JP2019515927A (ja) 感染症の処置のための方法
KR20150058159A (ko) 바클로펜 및 아캄프로세이트 기반 황반 변성 장애의 치료
US20230102036A1 (en) Use of compounds in the treatment of fungal infections
JP6799296B2 (ja) 新規プリオノイド病用治療薬
WO2020068562A1 (fr) Nouveaux traitements pour des infections amibiennes libres
TW202024058A (zh) 用於治療真菌感染之化合物及方法
RU2805930C1 (ru) Использование соединений при лечении грибковых инфекций
US20240082229A1 (en) Methods for preventing or treating h. pylori infection
US20230073499A1 (en) Compounds for use in the treatment of leukemia
WO2014064811A1 (fr) Agent thérapeutique de lutte contre l'hypertension pulmonaire
AU2013202360B2 (en) Methods of treatment using single doses of oritavancin
MX2015003810A (es) Metadoxina para usarse en el tratamiento de enfermedades hepaticas y formulaciones de liberacion prolongada de metadoxina.
WO2015016694A1 (fr) Composition solide stable d'immunosuppresseurs

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16751823

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 16751823

Country of ref document: EP

Kind code of ref document: A1