[go: up one dir, main page]

WO2024145451A2 - Méthodes de restauration du potentiel régénératif d'alvéoles pulmonaires âgées et de compartiments de cellules souches adultes âgées - Google Patents

Méthodes de restauration du potentiel régénératif d'alvéoles pulmonaires âgées et de compartiments de cellules souches adultes âgées Download PDF

Info

Publication number
WO2024145451A2
WO2024145451A2 PCT/US2023/086195 US2023086195W WO2024145451A2 WO 2024145451 A2 WO2024145451 A2 WO 2024145451A2 US 2023086195 W US2023086195 W US 2023086195W WO 2024145451 A2 WO2024145451 A2 WO 2024145451A2
Authority
WO
WIPO (PCT)
Prior art keywords
cells
subject
aged
inhibitor
young
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/US2023/086195
Other languages
English (en)
Other versions
WO2024145451A3 (fr
Inventor
Tuomas Tammela
Xueqian ZHUANG
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Memorial Sloan Kettering Cancer Center
Original Assignee
Memorial Sloan Kettering Cancer Center
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
Application filed by Memorial Sloan Kettering Cancer Center filed Critical Memorial Sloan Kettering Cancer Center
Publication of WO2024145451A2 publication Critical patent/WO2024145451A2/fr
Publication of WO2024145451A3 publication Critical patent/WO2024145451A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • 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
    • A61K31/427—Thiazoles 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
    • A61K31/54—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 sulfur as the ring hetero atoms, e.g. sulthiame
    • A61K31/5415—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 sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with carbocyclic ring systems, e.g. phenothiazine, chlorpromazine, piroxicam
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides

Definitions

  • the present disclosure generally relates to methods for restoring the regenerative potential of aged lung alveoli or aged adult stem cell compartments in a subject in need thereof comprising administering to the subject an effective amount of an inhibitor of NUPR1/LCN2-2 axis or iron/transferrin.
  • the subject is diagnosed with or suffers from a lung insufficiency such as COPD, COVID, influenza, and pneumonia sequelae.
  • methods for preventing or treating lung cancer in young (non-aged) subjects comprising administering an inhibitor of NUPR1/LCN2-2 axis, and/or ferroptosis inducing agent.
  • the present disclosure provides a method for restoring regenerative potential (sternness) of alveolar stem cells in a subject in need thereof comprising administering to the subject an effective amount of a Nuclear Protein 1 (NUPR1) inhibitor or a lipocalin-2 (LCN2) inhibitor.
  • NUPR1 Nuclear Protein 1
  • LPN2 lipocalin-2
  • the subject is diagnosed with or suffers from a lung insufficiency.
  • the present disclosure provides a method for preventing or treating lung cancer in a subject in need thereof comprising administering to the subject an effective amount of a Nuclear Protein 1 (NUPR1) inhibitor or a lipocalin-2 (LCN2) inhibitor, wherein the subject is a pediatric subject, an adolescent subject, or a young adult subject (e.g., ⁇ 55 years old).
  • a method for preventing or treating lung cancer in a subject in need thereof comprising administering to the subject an effective amount of a ferroptosis-inducing agent, wherein the subject is a pediatric subject, an adolescent subject, or a young adult subject (e.g., ⁇ 55 years old).
  • the young adult subject is about 54, about 53, about 52, about 51, about 50, about 49, about 48, about 47, about 46, about 45, about 44, about 43, about 42, about 41, about 40, about 39, about 38, about 37, about 36, about 35, about 34, about 33, about 32, about 31, about 30, about 29, about 28, about 27, about 26, about 25, about 24, about 23, about 22, about 21, or about 20 years old.
  • the ferroptosis-inducing agent may be a class 1 ferroptosis inducer (system X c “ inhibitor) or a class 2 ferroptosis inducer (glutathione peroxidase 4 (GPx4) inhibitor).
  • the LCN2 inhibitor is a small molecule, an LCN2-specific inhibitory nucleic acid, or an anti- LCN2 neutralizing antibody.
  • the LCN2 inhibitor is ZINC00784494, ZINC00640089, ZINC00230567, or ZINC00829534.
  • lung insufficiency examples include, but are not limited to, COPD (chronic obstructive pulmonary disease), cystic fibrosis, pneumonia, pulmonary embolism, COVID-19; conditions that affect the nerves and muscles that control breathing, such as amyotrophic lateral sclerosis (ALS), muscular dystrophy, spinal cord injuries, and stroke; problems with the spine, such as scoliosis (a curve in the spine); damage to the tissues and ribs around the lungs; drug or alcohol overdose; or inhalation injuries, such as from inhaling smoke (from fires) or harmful fumes.
  • the subject may be an immunocompromised subject or a geriatric subject.
  • FIG. IF Schematic representation of alveolar organoid formation assay (left). Primary and secondary alveolar organoid formation derived from aged vs. young primary AT2 cells (right).
  • the genes are ordered from most downregulated (dark blue) to the most upregulated (dark red) in aged across the x-axis.
  • Mean with SEM is shown in FIGs. 2A, 2B, 2E and 2F.
  • Y young; A: aged.
  • Student’s / test was used in FIGs. 2A-2C and
  • FIG. 3B Schematic summary of lentiviral Nuprl gene-targeting strategy in the context of LU AD tumorigenesis in vivo.
  • FIG. 3C Schematic summary of lentiviral Nuprl gene-targeting strategy in the context of LU AD tumorigenesis in vivo.
  • FIG. 5D Scatter plot showing mean differential methylation level of promoter regions (1 kb upstream and 200 bp downstream from the TSS) of the top 25 upregulated and downregulated genes among the aging-associated signature shared by both AT2 and LU AD cells (FIG. 2L). Pearson correlation was calculated. The regression line (blue) and its confidence interval (light grey) were plotted using a linear model. Genes transcriptionally upregulated in the aged AT2 and LU AD cells are shown in red; genes upregulated in the young are marked in blue.
  • FIG. 5E Experimental design of DNMT1 inhibition (DNMTl-i, orange) with GSK-3484862 over 8 days in vivo.
  • FIG. 51 Gene expression and promoter methylation change of genes among the top 25 upregulated aging signature genes (FIG.
  • FIG. 5K Contour plot showing the correlation between the fold change of DEGs in young transformed tumor spheres subjected to DNMTl-i or vehicle (x-axis) vs. aged or young vehicle controls (y-axis).
  • the grayscale represents the probability distribution. Pearson correlation was calculated.
  • the inset shows the percentage of genes in each quadrant, which is significantly different from random based on a binomial test (p ⁇ 2.2' 16 ).
  • the regression line (blue) and its confidence interval (light grey) were plotted using a linear model.
  • Mean with SD is shown in FIG. 5L.*/? ⁇ 0.05; **/? ⁇ 0.01; ***/? ⁇ 0.001.
  • One-way ANOVA was used in FIG. 5H; Student’
  • FIG. 6D Fold change in organoids formed by primary human AT2 cells in response to stimulation with recombinant human transferrin normalized to the number of control organoids. Scale bar: 50 gm.
  • FIG. 6E Correlation of young mouse LU AD and aged mouse LU AD gene expression signatures with patient age in The Cancer Genome Atlas data. All tumors had a KRAS mutation.
  • FIGs. 7A-7I Representative images of LU AD tumors in aged and young KPT (4 and 8 weeks) or KP (12 weeks) mice. At 4 and 8 weeks post-tumor initiation, tumor cells were visualized by tdTomato immunohistochemistry. Scale bar: 200 gm.
  • FIG. 7C FACS strategy for isolating mouse AT2 cells.
  • FIGs. 7G-7H Representative images (FIG. 7G) and quantification (FIG. 7H) of senescent cells identified by C12RG fluorescence-based detection of senescence-associated P-galactosidase activity.
  • FIG. 71 Quantification of lentiviral transduction efficiency of aged and young AT2 cells in the ex vivo transformation assay.
  • Mean with SD is shown in FIGs. 7D, 7F and 71.
  • Y young; A: aged. *p ⁇ 0.05; **p ⁇ 0.01; and ***p ⁇ 0.001. Student’s t test was used in FIGs. 7B, 7D, 7F, 7H and 71; one-way ANOVA was used in (e).
  • FIGs. 8A-8D Schematic summary of Eml4-Alk LU AD model (image representing chromosomal rearrangement reproduced from Maddalo et al. 34 ). Lungs of aged (104-130 weeks old) and young (12-16 weeks old) wild-type C57BL6/J mice were intratracheally transduced with adeno-sgEml4-sgAlk-Cas9 virus, which induces an Eml4-Alk gene fusion via an intra-chromosomal inversion on chromosome 17.
  • FIGs. 8B-8D Representative HE images (FIG. 8B) and LU AD tumor burden, quantified as tumor number/cross section (FIG.
  • FIGs. 9A-9D Quantification of senescent cancer cells in young vs. aged KP LU AD tumors at 12 weeks post-tumor initiation.
  • FIG. 9D Histopathological grading of KP LU AD tumors derived from aged vs. young mice at 12 and 17 weeks post-tumor initiation. Representative images of the grading by an automated deep neural network (Aiforia Technologies) are shown. Scale bar: 1 mm. Mean with SD is shown in FIG. 9B. Student’s t test was used in FIGs. 9B-9C.
  • FIG. 11C Violin plots showing expression of Lcn2 in young and aged AT2 cells and LU AD cell states. Rank-sum tests on single-cell gene expression vector were performed for significance testing.
  • FIGs. 12A-12H Density plot showing Enzymatic Methyl-Seq sequencing coverage in primary AT2 cell samples.
  • FIG. 12B Density plot of the methylation proportion in AT2 cell samples.
  • FIG. 12C Heatmap showing unsupervised clustering of aged vs. young AT2 cells based on DNA methylation profiles.
  • FIG. 12D Number of differentially methylated cytosines (DMCs) in aged vs. young AT2 cells based on location of CpG residues.
  • DMCs differentially methylated cytosines
  • FIG. 12E Density plot showing Enzymatic Methyl-Seq sequencing coverage in KP LU AD cell samples.
  • FIG. 12F Density plot of the methylation proportion of KP LU AD samples.
  • the present disclosure also provides an antisense nucleic acid comprising a nucleic acid sequence that is complementary to and specifically hybridizes with a portion of an NUPR1 mRNA or LCN2 mRNA, thereby reducing or inhibiting expression of NUPR1 or LCN2.
  • the antisense nucleic acid may be antisense RNA, or antisense DNA.
  • Antisense nucleic acids based on the known gene sequences of NUPR1 or LCN2 can be readily designed and engineered using methods known in the art.
  • the antisense nucleic acid comprises the nucleic acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or a complement thereof.
  • the antisense nucleic acid molecules may be administered to a subject or generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding the protein of interest to thereby inhibit expression of the protein, e.g., by inhibiting transcription and/or translation.
  • the hybridization can occur via Watson-Crick base pairing to form a stable duplex, or in the case of an antisense nucleic acid molecule which binds to DNA duplexes, through specific interactions in the major groove of the double helix.
  • NUPR1 or LCN2 mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel and Szostak (1993) Science 261 : 1411-1418, incorporated herein by reference.
  • the tracrRNA segment comprises a nucleotide sequence that is partially or completely complementary to the stem sequence of the crRNA and a nucleotide sequence that binds to the CRISPR enzyme.
  • the crRNA segment and the tracrRNA segment are provided as a single guide RNA.
  • the crRNA segment and the tracrRNA segment are provided as separate RNAs.
  • the combination of the CRISPR enzyme with the crRNA and tracrRNA make up a functional CRISPR-Cas system. Exemplary CRISPR-Cas systems for targeting nucleic acids, are described, for example, in WO20 15/089465.
  • the portion of the sequence 5' of the final “N” and upstream of the loop corresponds to the crRNA stem sequence
  • the portion of the sequence 3' of the loop corresponds to the tracrRNA sequence.
  • single polynucleotides comprising a guide sequence, a stem sequence, and a tracr sequence are as follows (listed 5' to 3'), where “N” represents a base of a guide sequence (e.g.
  • CRISPR enzymes are available for use in conjunction with the disclosed guide RNAs of the present disclosure.
  • the CRISPR enzyme is a Type II CRISPR enzyme.
  • the CRISPR enzyme catalyzes DNA cleavage.
  • the CRISPR enzyme catalyzes RNA cleavage.
  • the CRISPR enzyme is any Cas9 protein, for instance any naturally-occurring bacterial Cas9 as well as any chimeras, mutants, homologs or orthologs.
  • Cas proteins include Casl, CaslB, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8, Cas9 (also known as Csnl and Csxl2), CaslO, Csyl, Csy2, Csy3, Csel, Cse2, Cscl, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4, Csm5, Csm6, Cmrl, Cmr3, Cmr4, Cmr5, Cmr6, Csbl, Csb2, Csb3, Csxl7, Csxl4, CsxlO, Csxl6, CsaX, Csx3, Csxl, Csxl5, Csfl, Csf2, Csf3, Csf4, homologues thereof, or modified variants thereof.
  • a ferroptosis-inducing agent may be a class 1 ferroptosis inducer (system X c inhibitor) or a class 2 ferroptosis inducer (glutathione peroxidase 4 (GPx4) inhibitor).
  • the ferroptosis inducing agent is an inhibitory nucleic acid e.g., an antisense oligonucleotide, a shRNA, a sgRNA or a ribozyme) that targets GPX4.
  • an inhibitory nucleic acid e.g., an antisense oligonucleotide, a shRNA, a sgRNA or a ribozyme
  • Transferrin is a plasma glycoprotein capable of tightly but reversibly binding two atoms of iron and transporting them to proliferating cells for the synthesis of hemoglobin. Upon binding to the transferrin receptors on the cell surface, the transferrin is internalized within an endocytic vesicle. After the irons are dissociated from transferrin, irons pass through the endosomal membrane and enter into cytosol.
  • compositions of the present technology can be manufactured by methods well known in the art such as conventional granulating, mixing, dissolving, encapsulating, lyophilizing, or emulsifying processes, among others.
  • Compositions may be produced in various forms, including granules, precipitates, or particulates, powders, including freeze dried, rotary dried or spray dried powders, amorphous powders, tablets, capsules, syrup, suppositories, injections, emulsions, elixirs, suspensions or solutions.
  • Formulations may optionally contain solvents, diluents, and other liquid vehicles, dispersion or suspension aids, surface active agents, pH modifiers, isotonic agents, thickening or emulsifying agents, stabilizers and preservatives, solid binders, lubricants and the like, as suited to the particular dosage form desired.
  • the compositions disclosed herein are formulated for administration to a mammal, such as a human.
  • the rate of compound release can be controlled.
  • biodegradable polymers include poly(orthoesters) and poly(anhydrides).
  • Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.
  • Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or in a certain part of the intestinal tract, optionally, in a delayed manner.
  • Examples of embedding compositions that can be used include polymeric substances and waxes.
  • the active compounds can also be in micro-encapsulated form with one or more excipients as noted above.
  • the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch.
  • Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose.
  • the dosage forms may also comprise buffering agents.
  • opacifying agents may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or in a certain part of the intestinal tract, optionally, in a delayed manner.
  • embedding compositions include polymeric substances and waxes.
  • the present disclosure provides a method for restoring regenerative potential (sternness) of alveolar stem cells in a subject in need thereof comprising administering to the subject an effective amount of a Nuclear Protein 1 (NUPR1) inhibitor or a lipocalin-2 (LCN2) inhibitor.
  • NUPR1 Nuclear Protein 1
  • LPN2 lipocalin-2
  • the subject is diagnosed with or suffers from a lung insufficiency.
  • the present disclosure provides a method for preventing or treating lung cancer in a subject in need thereof comprising administering to the subject an effective amount of a Nuclear Protein 1 (NUPR1) inhibitor or a lipocalin-2 (LCN2) inhibitor, wherein the subject is a pediatric subject, an adolescent subject, or a young adult subject (e.g., ⁇ 55 years old). Also disclosed herein is a method for preventing or treating lung cancer in a subject in need thereof comprising administering to the subject an effective amount of a ferroptosis-inducing agent, wherein the subject is a pediatric subject, an adolescent subject, or a young adult subject (e.g., ⁇ 55 years old).
  • NUPR1 Nuclear Protein 1
  • LN2 lipocalin-2
  • ferroptosis-inducing agents include, but are not limited to, an inhibitory nucleic acid that targets GPX4, erastin, erastin derivatives (e.g., MEII, PE, AE, imidazole ketone erastin (IKE)), DPI2, BSO, SAS, lanperisone, SRS13-45, SRS13-60, RSL3, DPI7, DPI10, DPI12, DPI13, DPI17, DPI18, DPI19, ML160, sorafenib, artemisinin derivatives, artesunate, BAY87-2243, cisplatin, ironomycin, lanperisone, salinomycin, sulfasalazine, temozolomide, and lapatinib in combination with siramesine.
  • the lung cancer is lung adenocarcinoma (LU AD).
  • the lung cancer is lung adenocarcinoma (LU AD).
  • the NUPR1 inhibitor is a small molecule, an NUPR1 -specific inhibitory nucleic acid, or an anti-NUPRl neutralizing antibody.
  • the NUPR1 inhibitor is trifluoperazine (TFP), ZZW-115, ZZW-129, ZZW-130, ZZW-131, ZZW-132, ZZW-142, ZZW-143, ZZW-144, ZZW-145, or ZZW-148.
  • the LCN2 inhibitor is a small molecule, an LCN2-specific inhibitory nucleic acid, or an anti- LCN2 neutralizing antibody.
  • the LCN2 inhibitor is ZINC00784494, ZINC00640089, ZINC00230567, or ZINC00829534.
  • the NUPR1 -specific inhibitory nucleic acid, the LCN2-specific inhibitory nucleic acid or the inhibitory nucleic acid that targets GPX4 is a siRNA, a shRNA, an antisense oligonucleotide, or a sgRNA.
  • the present disclosure provides a method for restoring regenerative potential (sternness) of alveolar stem cells in a subject in need thereof comprising administering to the subject an effective amount of iron or transferrin.
  • the iron is unbound iron.
  • the iron is bound to transferrin or is complexed with vitamin C.
  • the subject is diagnosed with or suffers from a lung insufficiency.
  • lung insufficiency examples include, but are not limited to, COPD (chronic obstructive pulmonary disease), cystic fibrosis, pneumonia, pulmonary embolism, COVID-19; conditions that affect the nerves and muscles that control breathing, such as amyotrophic lateral sclerosis (ALS), muscular dystrophy, spinal cord injuries, and stroke; problems with the spine, such as scoliosis (a curve in the spine); damage to the tissues and ribs around the lungs; drug or alcohol overdose; or inhalation injuries, such as from inhaling smoke (from fires) or harmful fumes.
  • the subject may be an immunocompromised subject or a geriatric subject.
  • the present disclosure provides a method for restoring regenerative potential (sternness) of aged adult stem cell compartments in a subject comprising administering to the subject an effective amount of a Nuclear Protein 1 (NUPR1) inhibitor or a lipocalin-2 (LCN2) inhibitor.
  • NUPR1 Nuclear Protein 1
  • LPN2 lipocalin-2
  • the present disclosure provides a method for restoring regenerative potential (sternness) of aged adult stem cell compartments in a subject comprising administering to the subject an effective amount of iron or transferrin.
  • the aged adult stem cell compartments comprise one or more of neural adult subventricular zone (SVZ) stem cells, hair follicle stem cells, mesenchymal stem cells, keratinocyte stem cells, or intestinal stem cells.
  • SVZ neural adult subventricular zone
  • the iron is unbound iron. In other embodiments, the iron is bound to transferrin or is complexed with vitamin C.
  • the NUPR1 inhibitor, LCN2 inhibitor, ferroptosis-inducing agent, iron and/or transferrin is administered orally, topically, intranasally, systemically, intravenously, subcutaneously, intraperitoneally, intradermally, intraocularly, iontophoretically, transmucosally, or intramuscularly.
  • the subject is human.
  • the iron, transferrin, inhibitor of NUPR1/LCN2-2 axis, and/or ferroptosis inducing agent are administered to the subject in effective amounts (i.e., amounts that have desired therapeutic effect).
  • effective amounts i.e., amounts that have desired therapeutic effect.
  • the dose and dosage regimen will depend upon the degree of the disease symptoms in the subject, the characteristics of the iron, transferrin, inhibitor of NUPR1/LCN2-2 axis, and/or ferroptosis inducing agent, e.g., its therapeutic index, the subject, and the subject’s history.
  • the iron, transferrin, inhibitor of NUPR1/LCN2-2 axis, and/or ferroptosis inducing agent can be incorporated into pharmaceutical compositions for administration, singly or in combination, to a subject for the treatment or prevention of a disorder described herein.
  • Such compositions typically include the active agent and a pharmaceutically acceptable carrier.
  • pharmaceutically acceptable carrier includes saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.
  • Supplementary active compounds can also be incorporated into the compositions.
  • Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide.
  • a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents
  • antibacterial agents such as benzyl alcohol or methyl parabens
  • antioxidants
  • the iron, transferrin, inhibitor of NUPR1/LCN2-2 axis, or ferroptosis inducing agent described herein is administered by a parenteral route or a topical route.
  • Oral compositions generally include an inert diluent or an edible carrier.
  • the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules, e.g., gelatin capsules.
  • Oral compositions can also be prepared using a fluid carrier for use as a mouthwash.
  • Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition.
  • the iron, transferrin, inhibitor of NUPR1/LCN2-2 axis, and/or ferroptosis inducing agent of the present technology can be formulated in a carrier system.
  • the carrier can be a colloidal system.
  • the colloidal system can be a liposome, a phospholipid bilayer vehicle.
  • the therapeutic iron, transferrin, inhibitor of NUPR1/LCN2-2 axis, and/or ferroptosis inducing agent is encapsulated in a liposome while maintaining structural integrity.
  • there are a variety of methods to prepare liposomes. See Lichtenberg et al. , Methods Biochem.
  • Dosage, toxicity and therapeutic efficacy of the iron, transferrin, inhibitor of NUPR1/LCN2-2 axis, and/or ferroptosis inducing agent can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population).
  • the dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50/ED50.
  • an effective amount of the iron, transferrin, inhibitor of NUPR1/LCN2- 2 axis, and/or ferroptosis inducing agent range from about 0.000001 mg per kilogram body weight per day to about 10,000 mg per kilogram body weight per day.
  • the dosage ranges are from about 0.0001 mg per kilogram body weight per day to about 100 mg per kilogram body weight per day.
  • dosages can be 1 mg/kg body weight or 10 mg/kg body weight every day, every two days or every three days or within the range of 1-10 mg/kg every week, every two weeks or every three weeks.
  • the skilled artisan will appreciate that certain factors may influence the dosage and timing required to effectively treat a subject, including but not limited to, the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of the therapeutic compositions described herein can include a single treatment or a series of treatments.
  • the mammal treated in accordance present methods can be any mammal, including, for example, farm animals, such as sheep, pigs, cows, and horses; pet animals, such as dogs and cats; laboratory animals, such as rats, mice and rabbits. In some embodiments, the mammal is a human.
  • kits for restoring regenerative potential (sternness) of aged alveolar stem cells/aged adult stem cell compartments in a subject comprising one or more of iron, transferrin, or an inhibitor of NUPR1/LCN2-2 axis.
  • the subject is diagnosed with or suffers from a lung insufficiency, including but not limited to, chronic obstructive pulmonary disease (COPD), COVID, influenza, and pneumonia sequelae.
  • COPD chronic obstructive pulmonary disease
  • influenza influenza
  • pneumonia sequelae ae.
  • the aged adult stem cell compartments comprise one or more of neural adult subventricular zone (SVZ) stem cells, hair follicle stem cells, mesenchymal stem cells, keratinocyte stem cells, or intestinal stem cells.
  • SVZ neural adult subventricular zone
  • the above-mentioned components may be stored in unit or multi-dose containers, for example, sealed ampoules, vials, bottles, syringes, and test tubes, as an aqueous, preferably sterile, solution or as a lyophilized, preferably sterile, formulation for reconstitution.
  • the kit may further comprise a second container which holds a diluent suitable for diluting the pharmaceutical composition towards a higher volume. Suitable diluents include, but are not limited to, the pharmaceutically acceptable excipient of the pharmaceutical composition and a saline solution.
  • the kit may comprise instructions for diluting the pharmaceutical composition and/or instructions for administering the pharmaceutical composition, whether diluted or not.
  • the kit can also comprise, e.g., a buffering agent, a preservative or a stabilizing agent.
  • the kit can also contain a control sample or a series of control samples, which can be assayed and compared to the test sample.
  • Each component of the kit can be enclosed within an individual container and all of the various containers can be within a single package, along with instructions for interpreting the results of the assays performed using the kit.
  • the kits of the present technology may contain a written product on or in the kit container. The written product describes how to use the reagents contained in the kit. In certain embodiments, the use of the reagents can be according to the methods of the present technology.
  • Histological classification of mouse lung tumor grades was performed on the HE- stained sections by two methods: (i) an automated deep neural network (Aiforia Technologies, NSCLC_v25 algorithm), as and (ii) an independent classification by a board- certified veterinary pathologist, who was blinded to the sample group identifiers. Established histopathological criteria to evaluate mouse models of lung cancer were used 11,45 .
  • Tumor grades in pulmonary lobes ranged from 1 to 4 with grade 1 being composed by uniform histomorphology of the neoplastic cells without nuclear atypia and grade 4 being composed by pleomorphic neoplastic cells exhibiting increased nuclear atypia, mitoses and/or occasionally binucleated to multinucleated cells.
  • tumor subtypes were identified in the lungs of young and aged mice: solid adenomas and adenocarcinomas, papillary adenomas and adenocarcinoma, and mixed tumor subtypes containing both papillary and solid structures.
  • Immunohistochemistry was performed on 5 pm FFPE sections using standard staining protocols. Briefly, sections were de-paraffmized and heat-induced antigen retrieval was performed by EDTA antigen retrieval buffer (Sigma Aldrich, #E1161). Sections were blocked by BLOXALL solution (Vector laboratories, #SP-6000-100) at room temperature for 30 minutes and incubated with primary antibody at 4 °C overnight. IgG controls (Thermo Fisher Scientific, #02-6102, #02-6202 and #10400C) from the corresponding species of primary antibody were used as negative controls.
  • Immunofluorescence was performed on 5 pm FFPE sections. Briefly, sections were de-paraffinized and heat-induced antigen retrieval was performed by EDTA antigen retrieval buffer (Sigma Aldrich, #E1161). Sections were blocked by donkey immunomix [0.2% BSA (Sigma, #810533), 5% donkey serum (Thermo Fisher Scientific, #31874) and 0.3% Triton-X (Fisher Scientific, #BP151-100) in PBS (Gibco, #10010-023)] at room temperature for 30 minutes. Incubation of primary antibodies diluted in donkey immunomix was performed at 4 °C overnight.
  • IgG controls (Thermo Fisher Scientific, #02-6102, #02- 6202, and #10400C) from the corresponding species of primary antibody were used as negative controls.
  • AlexaFluor secondary antibodies raised in donkey were used for signal detection (Thermo Fisher Scientific #A-31571, #A-21207, #A32795, #A-11058, #A32787).
  • slides were counterstained with 1 pg/mL DAPI (Sigma Aldrich, #D9542) for 10 min, with coverslips using Mowiol mounting reagent (EMD Millipore, #475904). Mounted slides were digitally scanned by the Mirax Midi-Scanner (Carl Zeiss AG). Image analysis was performed by Fiji 53 .
  • FASTQ files of bulk-cell RNA-sequencing data were aligned to a custom GRCm38 / mmlO reference containing additional transgenes using STAR (version 2.7.5a) 65 .
  • Read counts were generated using the Python package HTSeq-count 66 .
  • the generated sample-gene count matrices were analyzed using a combination of published packages and custom scripts. The specific analytical workflows employed are summarized below.
  • Example 2 Aged AT2 cells exhibit reduced cell-intrinsic potential for proliferation and tumorigenesis
  • Example 6 DNA demethylation underpins aging-associated induction of the NUPR1- lipocalin-2 axis
  • DMCs differentially methylated cytosines
  • H3K4mel histone-3
  • H3K27ac histone-3
  • H3K4me3 modifications 40 FIG. 13A.
  • Example 7 Aging-induced changes in AT2 cells and LU AD are conserved in humans
  • Example 8 Young AT2 cells are sensitive to ferroptosis but aged cells are resistant
  • Example 9 Aging induces the NUPRl-lipocalin-2 axis in other adult stem cell compartments
  • a range includes each individual member.
  • a group having 1-3 cells refers to groups having 1, 2, or 3 cells.
  • a group having 1-5 cells refers to groups having 1, 2, 3, 4, or 5 cells, and so forth.
  • HTSeq— 1228 a Python framework to work with high throughput sequencing data. Bioinformatics 31, 166-169 (2015). Love, M. I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 15, 550 (2014). Krueger, F. & Andrews, S. R. Bismark: a flexible aligner and methylation caller for Bisulfite-Seq applications. Bioinformatics 27, 1571-1572 (2011). Krueger, F. Trim Galore. (2022). Martin, M. Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet.journal 17 (2022). Langmead, B.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pulmonology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente divulgation concerne des méthodes de restauration du potentiel de régénération d'alvéoles pulmonaires âgées ou de compartiments de cellules souches adultes âgés chez un sujet en ayant besoin, comprenant l'administration au sujet d'une quantité efficace d'un inhibiteur de l'axe NUPR1/LCN2-2 ou du fer/transferrine. Dans certains modes de réalisation, le sujet est diagnostiqué comme présentant une insuffisance pulmonaire, ou est atteint d'une insuffisance pulmonaire, telle que les séquelles de la BPCO, de la COVID, de la grippe et de la pneumonie. Sont également divulguées des méthodes de prévention ou de traitement du cancer du poumon chez des sujets jeunes (non âgés) comprenant l'administration d'un inhibiteur de l'axe NUPR1/LCN2-2, et/ou d'un agent induisant la ferroptose.
PCT/US2023/086195 2022-12-29 2023-12-28 Méthodes de restauration du potentiel régénératif d'alvéoles pulmonaires âgées et de compartiments de cellules souches adultes âgées Ceased WO2024145451A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202263435914P 2022-12-29 2022-12-29
US63/435,914 2022-12-29

Publications (2)

Publication Number Publication Date
WO2024145451A2 true WO2024145451A2 (fr) 2024-07-04
WO2024145451A3 WO2024145451A3 (fr) 2024-07-25

Family

ID=91719248

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2023/086195 Ceased WO2024145451A2 (fr) 2022-12-29 2023-12-28 Méthodes de restauration du potentiel régénératif d'alvéoles pulmonaires âgées et de compartiments de cellules souches adultes âgées

Country Status (1)

Country Link
WO (1) WO2024145451A2 (fr)

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006078717A2 (fr) * 2005-01-19 2006-07-27 Beth Israel Deaconess Medical Center Lipocaline 2 pour traiter, prevenir et maitriser les metastases cancereuses, l'angiogenese et la fibrose
EP2771014A4 (fr) * 2011-10-28 2015-07-08 Univ Nat Yang Ming Composition pharmaceutique destinée à éradiquer les cellules souches cancéreuses
US9351949B2 (en) * 2012-07-19 2016-05-31 Sbi Pharmaceuticals Co., Ltd. Prophylactic/therapeutic agent for influenza virus infection
ES2774330T3 (es) * 2013-04-09 2020-07-20 Massachusetts Inst Technology Polímero de suministro de fármaco y usos del mismo
HUE070063T2 (hu) * 2016-03-15 2025-05-28 Solvotrin Therapeutics Ltd Készítmények és módszerek a vasbevitel növelésére emlõsök esetében
US11406664B2 (en) * 2016-03-31 2022-08-09 Shield TX (UK) Limited Ferric maltol compositions for use in the treatment or prevention of cancer and tumours
CN113905762A (zh) * 2019-05-30 2022-01-07 北京生命科学研究所 特发性肺纤维化的药物靶点
WO2021105384A1 (fr) * 2019-11-27 2021-06-03 INSERM (Institut National de la Santé et de la Recherche Médicale) Ciblage de la région nls de la protéine nupr1 pour traiter le cancer
US11768206B2 (en) * 2020-03-21 2023-09-26 Henry Winchester Methods of treating or preventing serious symptoms from COVID-19 infection
EP3892275A1 (fr) * 2020-04-08 2021-10-13 INSERM (Institut National de la Santé et de la Recherche Médicale) Production d'aérosols de hcq ou de ses métabolites pour le traitement d'infections pulmonaires
WO2021222660A1 (fr) * 2020-04-30 2021-11-04 Kaleido Biosciences, Inc. Compositions d'oligosaccharides et leurs procédés d'utilisation pour le traitement d'infection virales
CN115916213A (zh) * 2020-05-28 2023-04-04 胡建倧 用于治疗SARS-CoV-2的组合物和方法

Also Published As

Publication number Publication date
WO2024145451A3 (fr) 2024-07-25

Similar Documents

Publication Publication Date Title
Hu et al. Oncogenic KRAS signaling drives evasion of innate immune surveillance in lung adenocarcinoma by activating CD47
Ciesla et al. Heme oxygenase-1 controls an HDAC4-miR-206 pathway of oxidative stress in rhabdomyosarcoma
JP6081798B2 (ja) ｍｉＲＮＡに関連する癌を検出および処置するための方法および組成物およびｍｉＲＮＡインヒビターおよび標的
US11490603B2 (en) Animal model of brain tumor and manufacturing method of animal model
US20220389393A1 (en) Compositions and methods for editing of the cdkl5 gene
US20100137411A1 (en) Ras-mediated epigenetic silencing effectors and uses thereof
US11814688B2 (en) Markers for determining tumor hypoxia
JP6566933B2 (ja) ガンを治療するための療法のための標的として使用するためのｆａｌｚ
EP3371211B1 (fr) Inhibiteur de protéine kinase associée à rho dans le traitement de tumeurs et du cancer, et identification de sujets candidats à ce traitement
WO2024229094A1 (fr) Procédés de traitement de maladies inflammatoires gastro-intestinales à l'aide de modulateurs d'alpha-cétoglutarate (akg)
Chen et al. EGR3 Inhibits Tumor Progression by Inducing Schwann Cell‐Like Differentiation
US20220259673A1 (en) Methods for identifying and treating high-plasticity cell state driving tumor progression in lung cancer
WO2020051342A1 (fr) Méthodes pour traiter une maladie métastatique faisant appel à un inhibiteur cx 5461 de la biogenèse des ribosomes
EP4014047B1 (fr) Inhibiteurs d'ogdh pour utilisation dans la traitement de cancer
US20210220471A1 (en) Methods of using pharmacologic inhibitors of type 2 cytokine signaling to treat or prevent pancreatic cancer
WO2024145451A2 (fr) Méthodes de restauration du potentiel régénératif d'alvéoles pulmonaires âgées et de compartiments de cellules souches adultes âgées
US11266677B2 (en) Methods for treatment or prevention of leukemia
US11246868B2 (en) Treatment of hippo pathway mutant tumors and methods of identifying subjects as candidates for treatment
Liu et al. ALKB homolog 5 (ALKBH5)-induced circPUM1 upregulation facilitated the progression of neuroblastoma via miR-423–5p/PA2G4 axis
CA3202058A1 (fr) Agent de ciblage jmjd6 pour traiter le cancer de la prostate
CN112725436A (zh) 一种人circMKLN1基因的用途及相关产品
US20230232794A1 (en) Animal model of brain tumor and manufacturing method of animal model
Wu et al. CircSP3 promotes ccRCC progression and sunitinib resistance via encoding SP3-461aa to stabilize MYH9 and activate the PI3K-AKT pathway
Cai Translational mechanisms of stem cell fate regulation in epidermal oncogene tolerance
JP7097606B2 (ja) 二本鎖核酸分子、ｄｎａ、ベクター、がん細胞増殖抑制剤、医薬、及びｓｐｏｎ１－ｔｒｉｍ２９融合遺伝子の利用

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: 23913717

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase

Ref country code: DE