WO2020036658A2 - Médicaments favorisant la survie des photorécepteurs du bâtonnet rétinien - Google Patents

Médicaments favorisant la survie des photorécepteurs du bâtonnet rétinien Download PDF

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WO2020036658A2
WO2020036658A2 PCT/US2019/029258 US2019029258W WO2020036658A2 WO 2020036658 A2 WO2020036658 A2 WO 2020036658A2 US 2019029258 W US2019029258 W US 2019029258W WO 2020036658 A2 WO2020036658 A2 WO 2020036658A2
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photoreceptor
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retinal
yfp
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Jeff Mumm
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Johns Hopkins University
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    • 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
    • A61K31/4418—Non condensed pyridines; Hydrogenated derivatives thereof having a carbocyclic group directly attached to the heterocyclic ring, e.g. cyproheptadine
    • 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
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/357—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
    • 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
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
    • A61K31/37—Coumarins, e.g. psoralen
    • 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
    • A61K31/4402—Non condensed pyridines; Hydrogenated derivatives thereof only substituted in position 2, e.g. pheniramine, bisacodyl
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A61K33/30—Zinc; Compounds thereof

Definitions

  • HTS target-directed high- throughput screening
  • a combination of compounds may be administered to a subject.
  • An example of a combined therapy includes administering dhydroartemisinin, artemisinin, and ciclopirox olamine to a subject.
  • the one or more compounds of Figs. 7 and 13 may administered to a subject by any suitable means such as eye drops delivering one or more compounds to the eye of a subject.
  • a pharmaceutical composition of the one or more compounds of Figs. 7 and 13 may be formed prior to the administration to a subject.
  • Another embodiment of the present invention is a method of promoting photoreceptor and/or retinal cell survival.
  • the method includes the steps of: administering one or more compounds of Figs. 7 and 13 to a photoreceptor, a retinal cell, or both having a disease or treated with an agent that causes cell death; and promoting the survival of the photoreceptor, retinal cell, or both compared to a reference photoreceptor, a reference retinal cell, or both that have not been administered one or more compounds of Figs 7 and 13.
  • the photoreceptor, the retinal cell, the reference photoreceptor, and the reference retinal cell used may have a disease, causing blindness such as RP as an example, prior to administering one or more compounds of Figs. 7 and 13.
  • Other embodiments of the present invention use a photoreceptor, a retinal cell, or both that is treated with the agent that causes photoreceptor cell death, retinal cell death, or both after the administering of one or more compounds of Figs. 7 and 13.
  • Suitable the agents used in the present invention tunicamycin, thapsigargin or a combination thereof, as examples.
  • Another embodiment of the present invention includes a method of treating or preventing blindness in a subject.
  • the method includes the steps of: administering to a subject having a disease resulting in photoreceptor death, retinal cell death, or a combination thereof, one or more compounds of Figs. 7 and 13; and treating or preventing blindness of the subject compared to a reference subject that has not been administered the one or more compounds of Figs. 7 and 13.
  • agent any small molecule chemical compound, antibody, nucleic acid molecule, or polypeptide, or fragments thereof.
  • ameliorate is meant decrease, suppress, attenuate, diminish, arrest, or stabilize the development or progression of a disease.
  • analog is meant a molecule that is not identical, but has analogous functional or structural features.
  • disease is meant any condition or disorder that damages or interferes with the normal function of a cell, tissue, or organ.
  • diseases include retinitis pigmentosa (RP).
  • an effective amount is meant the amount required to ameliorate the symptoms of a disease relative to an untreated patient.
  • the effective amount of active compound(s) used to practice the present invention for therapeutic treatment of a disease varies depending upon the manner of administration, the age, body weight, and general health of the subject.
  • Ranges provided herein are understood to be shorthand for all of the values within the range.
  • a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,
  • the terms“treat,” treating,”“treatment,” and the like refer to reducing or ameliorating a disorder and/or symptoms associated therewith. It will be appreciated that, although not precluded, treating a disorder or condition does not require that the disorder, condition or symptoms associated therewith be completely eliminated.
  • the terms“prevent,”“preventing,”“prevention,”“prophylactic treatment” and the like refer to reducing the probability of developing a disorder or condition in a subject, who does not have, but is at risk of or susceptible to developing a disorder or condition.
  • Figure 1A-1B Primary Screen.
  • A. Rod photoreceptor ablation at different Mtz concentrations. Five concentrations of Mtz from 10 mM to 0.625 mM were tested for the ability to induce rod photoreceptor ablation in rho:YFP-NTR larvae using a 48 hr treatment regimen from 5 to 7 dpf (days post-fertilization), ARQiv quantification of YFP loss, and a sample size of 24 per condition. The 2.5 mM Mtz treated group displayed progressive YFP loss, equivalent to higher concentrations by the second day of treatment, and was therefore chosen as the regimen for large-scale screening.
  • YFP signal of each larva was quantified by ARQiv (i.e. Tecan Ml 000 Pro microplate reader assay).
  • ARQiv i.e. Tecan Ml 000 Pro microplate reader assay.
  • Data was collected and analyzed in near real-time to allow same day visual confirmation of compound effects.
  • Data processing included a signal to background ratio plot, a SSMD score plot, a heat map of signal to background ratio within the plate, and SSMD score table.
  • Conditions producing an SSMD score >1 were visually inspected using stereo fluorescence microscopy. Conditions producing elevated YFP signals either from drug autofluorescence or deceased larvae were excluded.
  • FIG. 3 Intravital Imaging. Confocal z-stack projections images show YFP reporter signals in rho:YFP-NTR transgenic larval zebrafish eyes following Mtz-induced rod photoreceptor loss and treatment with hit compounds or vehicle control. At 7 dpf, non- ablated retinas displayed strong YFP expression in rod photoreceptors in the outer nuclear layer (ONL) with elevated signals in the ventral retina corresponding to a higher of rod cells in this region. Mtz-treated rho:YFP-NTR retinas showed reduced YFP levels when treated with 0.1% DMSO vehicle alone (negative control). Retinas treated with hit compounds exhibited enhanced YFP signals relative to the negative control, thus preservation of rod photoreceptor cells. High magnification images of hit compound treated retinas detail surviving rod cell morphologies, which ranged from healthy elongated rod cell structure to condensed cells, suggesting variable effects on preservation of rod photoreceptor function.
  • FIG. 4 Cell Death Assay. TUNEL staining was used to quantify cell death. Non- ablated control retinas showed only occasional TUNEL-positive cells. In retinas treated with 2.5 mM Mtz, YFP signal was lost and an increased number of TUNEL-positive cells was observed in the ONL. Mtz-treated retinas exposed to hit compounds, e.g., dihydroartemisin (Dih), showed sustained YFP signals and a reduction in TUNEL-positive cells, suggesting suppression of cell death. The mean, SD and sample size of each group is listed. Student’s t- test was used to calculate the p value.
  • hit compounds e.g., dihydroartemisin (Dih)
  • Additive Effects Assay Combinatorial assays pairing 7 hit compounds at their maximal effective concentrations were used to test for additive effects. Ten pairs produced additive effect sizes (all green boxes) near or above the sum of their individual effects (italicized values in lower left). Moreover, eight pairs produced better than additive effects (dark green boxes).
  • Figure 7. Eleven hit compounds abbreviations.
  • Figure 8. Mouse Primary Retinal Cell Culture Assays. Table summarizing protective effects of tested hit compounds on primary cultures of isolated mouse retinal cells treated with exogenous stressor compounds to induce cell death. (++: >2SD; +++: >3SD)
  • NTR Inhibition Assay The effect of 11 hit compounds on NTR enzymatic activity was tested. NTR activity was estimated using CB1954 kinetics assay in the presence of test compounds at 300 mM. The NTR activity in each compound treated sample was compared to vehicle only controls. If the ratio was higher than 0.75, drugs were not considered to have an inhibitory effect. War, Cic, Cal and Sul were found to inhibit NTR activity at 300 mM. Using the same drug concentrations, NTR inhibitory effects of War, Cic, Cal was also tested using an Mtz kinetics assay. Subsequently, IC50 values were calculated. Relative NTR activity levels, standard deviation and IC50 in CB1954 kinetics assay, as well as NRT activity in Mtz kinetics assay are summarized in the table.
  • FIG. 10 Rod Cell Fate Assay. Effects of 11 hit compounds on rod cell differentiation was tested. Larvae were treated with hit compounds at maximal
  • Mtz was then rinsed out and larvae exposed to hit compound for 3 days (6 to 9 dpf).
  • YFP signal was quantified at 9 dpf. All signals were compared to Mtz ablated controls treated with vehicle alone. Non-ablated larvae served as positive controls. None of the larvae treated with the hit compounds showed improved YFP signals compared to vehicle controls; two compounds (Clo and Cic) showed reduced YFP signals (black asterisks, p-value ⁇ 0.005). Student’s /-test with multiple comparisons correction was performed to calculate 95% confidence intervals and p values, shown in a table below the graph along with the sample size for each compound.
  • FIG. 14 Possible signaling pathways for each hit compound. Searches were conducted in in PubChem (https://puhchem.ncbi.nlm.nih.gov/) to assess bioassay results for hit compounds, and previously implicated signaling pathways are summarized. Note that some drugs appear to modulate multiple pathways and many of the implicated signaling pathways overlap between hit compounds. For example, Tdpl inhibition was shared by 8 out of 11 hit compounds.
  • Tdpl Tyrosyl-DNA phosphodiesterase 1; Rorc: RAR related orphan receptor gamma; AR: androgen receptor signaling pathway; TR: thyroid receptor signaling pathway; VDR: vitamin D receptor; ER: estrogen receptor alpha signaling; AhR: aryl hydrocarbon receptor signaling; GR: glucocorticoid receptor; Dopa: dopamine related; HIF1 : Hypoxia-inducible factor 1 -alpha; SHH: Sonic hedgehog; COX: cyclooxygenase.
  • Zebrafish are amendable to large-scale chemical screening due to their high fecundity rate, small size, and ease of visualizing phenotypes across gross morphological, tissue- specific, cell-specific, molecular, and behavioral levels of analysis (Mathias et al., 2012) (Zon and Peterson, 2005).
  • Semi-automated low- to mid-throughput compound screens have been performed with zebrafish using high-content imaging (Selderslaghs et al, 2009)(Padilla et al, 20l2)(Ton et al, 2006) or behavioral assay platforms (Rihel et al., 20l0)(Kokel et al, 2010).
  • the inventors established a complementary platform, automated reporter quantification in vivo (ARQiv; (Walker et al, 20l2)(White et al, 2016)) to enable high-throughput screening rates in whole-organism (Wang et al, 2015) and organoid (Vergara et al, 2017) disease models.
  • ARQiv automated reporter quantification in vivo
  • the ARQiv system was used to perform a large-scale whole-organism screen to identify potential new therapeutics for delaying photoreceptor loss in RP patients. Using an inducible zebrafish model of RP, over 300,000 larvae were screened to identify
  • Neuroprotective compounds that promoted rod photoreceptor survival.
  • a collection of approximately 3,000 largely human-approved compounds (the Johns Hopkins Drug Library; (Shim and Liu, 2014), was evaluated across six concentrations using quantitative HTS (qHTS; (Inglese et al, 2006b)) principles.
  • the inventors identified 114 hits with and 42 of the top performing compounds were advanced through a series of confirmatory and orthogonal assays. Eleven compounds passed all secondary tests and moved forward as lead drug candidates.
  • YFP-NTR larvae were treated with either 2.5mM Mtz/0. l% DMSO or 0.1% DMSO (control) from 5-7 dpf. Each group contained at least 48 larvae and three biological replicates were performed. YFP signals were measured using the TECAN microplate reader at 7 dpf. Power calculations were used to determine sample sizes across a range of error rates and effect sizes for both raw and log2 -transformed data, as per published methods (White et al, 2016). A sample size of 9 per condition was predicted to facilitate detection of an effect size of 50% of non-ablated controls with error rates of 0.05 for false-positives and false-negatives (type I and type II, respectively). However, to account for dispensing errors and in keeping with 96-well microtiter plate formats, a sample size of 16 was chosen for the primary screen.
  • JHDL John Hopkins Drug Library
  • the JHDL consists of -2,200 human-approved drugs and an additional -800 in clinical trials (Chong et al, 2006).
  • the ARQiv-based screening process has been detailed previously (White et al, 2016) and adapted here for large-scale quantification of YFP-expressing rod photoreceptors.
  • rho:YFP-NTR embryos were collected and raised in zebrafish E3 embryo media (5 mM NaCl; .17 mM KC1; .33 mM CaCl; .33 mM MgS04).
  • E3 media E3/PTU
  • visual screens were performed to remove larvae with abnormal morphology or low retinal YFP levels.
  • Stock drug and DMSO (negative control) solutions were automatically dispensed into and diluted across a 96-well plate containing E3/PTU using a robotic liquid handling system (Hudson Robotics), as previously described (White et al, 2016).
  • a COPAS-XL Complex Object Parametric Analyzer and Sorter, Union Biometrica
  • a COPAS-XL Complex Object Parametric Analyzer and Sorter, Union Biometrica
  • YFP signals were measured using TECAN Ml 000 PRO microplate reader (excitation 514 nm, bandwidth 5 nm; emission 538nm, bandwidth 10 nm; note these settings are optimized for the eYFP reporter present in the rho:YFP-NTR transgene, and different than previously published settings for tag YFP, (Wang et al, 2015) (White et al, 2016)). Larvae exposed to 2.5mM Mtz but treated only with 0.1% DMSO served as controls for maximal rod photoreceptor cell ablation.
  • top-performing‘hit’ compounds were selected for confirmatory and orthogonal assays.
  • hit drugs were obtained from new sources and tested across a wider range of concentrations, using a five-fold dilution series and a total of eight concentrations, with a sample size 30 fish per condition. Based on the toxicity profile of each drug, the starting concentration was either 100, 10 or 1 mM. Three biological replicates were conducted. The results were normalized and pooled to calculate effect sizes, confidence intervals, and p-values using Student’s t-test.
  • non-ablated (positive control), 2.5mM Mtz- ablated (negative control) and Mtz+drug treated rho:YFP-NTR larvae were collected at 7 dpf (i.e., after a 48 hr exposure as per the primary screen). All compounds were tested at the maximal effective concentration.
  • Three larvae from each group were embedded in 1% low melt agarose gel with either the left or right eye facing up.
  • An Olympus Fluoview FV1000 confocal microscope with a 20x water immersion objective (0.95 NA) was used to take 30-40 images at 4 pm intervals. These image stacks were then processed to produce a maximal intensity projection of the entire retina.
  • a region in the dorsal -nasal quadrant was imaged with a 60x water immersion objective (1.10 NA) to provide greater detail of rod
  • rho:YFP-NTR larvae were handled as described for the primary screen with the following exceptions. At 5 dpf, larvae were dispensed into 96-well plate and exposed solely to tested compounds; larvae were not treated with Mtz. YFP reporter signals were then quantified by microplate reader at 7 dpf. Each experiment was repeated at least twice, with tested groups containing
  • both non-transgenic and transgenic rho:YFP-NTR larvae were treated with PTU at 16 hpf.
  • larvae were incubated with either lOmM Mtz/E3/TU or DMSO/E3/PTU for 24 hours.
  • larvae were then placed in new DMSO/E3/PTU media containing test compounds (or DMSO alone) for three days.
  • YFP signal intensity was measured by TECAN microplate reader at 9 dpf. Each experiment was repeated at least twice, with tested groups containing approximately 30 fish.
  • Eyes were then incubated in 1 mL of high glucose Hanks balanced salt solution containing 0.5mg/mL proteinase K at 37°C for 7 min.
  • the eyeballs were then placed in Neurobasal medium (Life Technologies) plus 10% fetal calf serum to stop enzymatic activity.
  • the retina with attached RPE attached was dissected free from the choroid and sclera. Relaxing cuts were made to flatten the tissue and transferred to the upper compartment of a Costar Transwell chamber using a drop of Neurobasal medium, RPE layer faced-down. A drop of fluid was used to flatten-out the retina.
  • Neurobasal media with B-27 supplement (Life Technologies) was placed in the lower compartment. The cultures were kept in an incubator (5% C02, balanced air, 100% humidity, 37°C) and the media in the lower compartment changed every two days under dim red light. No antimitotics nor antibiotics were required.
  • the ARQiv data analysis package was used to calculate sample size, quality control strictly standardized mean difference (SSMD) and hit selection SSMD scores as previously described (White et al, 2016).
  • the following results of each drug were derived: 1) a plot of signal: background at all tested concentrations, 2) at table of SSMD hit selection scores, and 3) a signal intensity heat map of each drug plate. (96-well plate view).
  • a transgenic line, Tg(rho:YFP-Eco.NfsB)gmc500 (hereafter, rho:YFP-NTR ), facilitating prodrug-inducible rod photoreceptor loss (Walker et al, 2012).
  • rho 3.7kb rhodopsin (rho) promoter fragment
  • YFP yellow fluorescent protein
  • NfsB bacterial nitroreductase
  • ARQiv could be used to quantify loss and regeneration kinetics of rod photoreceptors following a 24 hr exposure of lOmM Mtz from 5-6 dpf (Walker et al, 2012).
  • confocal intravital imaging suggested maximal loss of YFP occurred two days after initiation of Mtz treatments (Walker et al, 2012). Consequently, a 48 hr Mtz exposure regimen would provide maximal YFP (rod photoreceptor) loss, thereby providing the broadest signal window to detect neuroprotective effects. Concluding the experiment at 7 dpf also avoids complications associated with exogenous feeding (see Walker et al.
  • ARQiv was used to quantify changes in YFP reporter signal daily from 5-8 dpf.
  • the data showed a concentration-dependent reduction in YFP reporter signal for all treatment regimens.
  • Maximal YFP loss was observed at 7 dpf with 10, 5, and 2.5mM Mtz exposures (Figure 1 A).
  • the 2.5mM Mtz treatment in particular, displayed a linear decrease in YFP levels from 5 to 7 dpf, consistent with gradual rod photoreceptor loss (Figure 1A).
  • no evidence of toxicity or deleterious effects on morphology were observed for 48 hr Mtz treatments of 2.5 mM or less.
  • XL880 also affected general morphology (e.g., causing edema around the eye) thus making it unclear whether sustained YFP levels resulted from rescued rod photoreceptors or changes in detection of the reporter associated with edema. Nevertheless, as no other previously implicated neuroprotectant proved effective in our model, XL880 was included as a positive control for the detection of increased YFP signals in the primary screen.
  • NTR prodrug converting enzyme NTR prodrug converting enzyme
  • rho:YFP-NTR larvae were exposed to hit drugs alone (i.e. no Mtz induced cell ablation) from 5 to 7 dpf and YFP levels evaluated by ARQiv.
  • Retinoic acid (RA, 1.25 mM) was used as a positive control.
  • RA treated fish displayed significantly increased YFP signals (Fig. 10).
  • none of the eleven hit compound treated retinas exhibited increased YFP expression compared to untreated controls, suggesting that the compounds do not promote rod photoreceptor cell fate (Fig. 10).
  • three hit compounds produced reproducibly lower YFP signals than controls (Clo, Cic, and Cor; p-value ⁇ 0.05), suggesting negative effects on rod photoreceptor development.
  • orthologous human gen & PDE6B also leads to RP in humans (McLaughlin et al, l993)(Gal et al, l994)(Bayes et al, l995)(Hmani-Aifa et al, 2009).
  • photoreceptor degeneration begins around P10 and by P21, only a few rows of photoreceptor cells remain in the ONL (LaVail and Sidman, 1974).
  • retinal explants from P10 rdl mice were isolated and cultured ex vivo as previously described (Bandy opadhyay and Rohrer, 2010). In the absence of exogenous factors, photoreceptor degeneration proceeds rapidly under these conditions.
  • TUNEL terminal deoxynucleotidyl transferase dUTP nick end labeling
  • Combinatorial assays demonstrated additive effects of -50% tested pairs, indicating that complementary signaling pathways regulate cell death during retinal degeneration.
  • retinal cell death is characterized by the excessive activation of histone deacetylase (HD AC), poly-ADP-ribose-polymerase (PARP), calpain and the abnormal accumulation of cyclic guanosine monophosphate (cGMP) and poly-ADP-ribose (PAR).
  • HD AC histone deacetylase
  • PARP poly-ADP-ribose-polymerase
  • calpain the abnormal accumulation of cyclic guanosine monophosphate (cGMP) and poly-ADP-ribose (PAR).
  • Embodiments of the disclosure concern methods and/or compositions for treating and/or preventing Retinitis pigmentosa (RP).
  • individuals with Retinitis pigmentosa are administered with one or more compounds of Figs. 7 and 13.
  • an individual is given an agent for RP therapy in addition to the one or more compounds of Figs. 7 and 13.
  • the additional therapy may be given prior to, at the same time as, and/or subsequent to the one or more compounds of Figs. 7 and 13.
  • compositions of the present invention comprise an effective amount of one or more the one or more compounds of Figs. 7 and 13, dissolved or dispersed in a
  • pharmaceutically acceptable carrier refers to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to an animal, such as, for example, a human, as appropriate.
  • the preparation of a pharmaceutical composition that comprises at least one or more compounds of Figs. 7 and 13 or additional active ingredient will be known to those of skill in the art in light of the present disclosure, as exemplified by Remington: The Science and Practice of Pharmacy, 21 st Ed. Lippincott Williams and Wilkins, 2005, incorporated herein by reference.
  • preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biological Standards.
  • FDA Office of Biological Standards As used herein,
  • “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, drugs, drug stabilizers, gels, binders, excipients, disintegration agents, lubricants, sweetening agents, flavoring agents, dyes, such like materials and combinations thereof, as would be known to one of ordinary skill in the art (see, for example, Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990, pp. 1289-1329, incorporated herein by reference).
  • preservatives e.g., antibacterial agents, antifungal agents
  • isotonic agents e.g., absorption delaying agents, salts, preservatives, drugs, drug stabilizers, gels, binders, excipients, disintegration agents, lubricants, sweetening agents, flavoring agents, dye
  • the one or more compounds of Figs. 7 and 13 may comprise different types of carriers depending on whether it is to be administered in solid, liquid or aerosol form, and whether it need to be sterile for such routes of administration as injection.
  • the one or more compounds of Figs. 7 and 13 may be formulated into a composition in a free base, neutral or salt form.
  • Pharmaceutically acceptable salts include the acid addition salts, e.g., those formed with the free amino groups of a proteinaceous composition, or which are formed with inorganic acids such as for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric or mandelic acid. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as for example, sodium, potassium, ammonium, calcium or ferric hydroxides; or such organic bases as
  • solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective.
  • the formulations are easily administered in a variety of dosage forms such as formulated for parenteral administrations such as injectable solutions, or aerosols for delivery to the lungs, or formulated for alimentary administrations such as drug release capsules and the like.
  • the composition of the present invention suitable for administration is provided in a pharmaceutically acceptable carrier with or without an inert diluent.
  • the carrier should be assimilable and includes liquid, semi-solid, i.e., pastes, or solid carriers. Except insofar as any conventional media, agent, diluent or carrier is detrimental to the recipient or to the therapeutic effectiveness of a composition contained therein, its use in administrable composition for use in practicing the methods of the present invention is appropriate.
  • carriers or diluents include fats, oils, water, saline solutions, lipids, liposomes, resins, binders, fillers and the like, or combinations thereof.
  • composition may also comprise various antioxidants to retard oxidation of one or more component. Additionally, the prevention of the action of microorganisms can be brought about by preservatives such as various antibacterial and antifungal agents, including but not limited to parabens (e.g., methylparabens, propylparabens), chlorobutanol, phenol, sorbic acid, thimerosal or combinations thereof.
  • parabens e.g., methylparabens, propylparabens
  • chlorobutanol phenol
  • sorbic acid thimerosal or combinations thereof.
  • the composition is combined with the carrier in any convenient and practical manner, i.e., by solution, suspension, emulsification, admixture, encapsulation, absorption and the like. Such procedures are routine for those skilled in the art.
  • the present invention may concern the use of a
  • lipid vehicle compositions that include the one or more compounds of Figs. 7 and 13, and an aqueous solvent.
  • the term“lipid” will be defined to include any of a broad range of substances that is characteristically insoluble in water and extractable with an organic solvent. This broad class of compounds are well known to those of skill in the art, and as the term“lipid” is used herein, it is not limited to any particular structure. Examples include compounds which contain long-chain aliphatic hydrocarbons and their derivatives. A lipid may be naturally occurring or synthetic (i.e., designed or produced by man). However, a lipid is usually a biological substance.
  • the one or more compounds of Figs. 7 and 13 may be dispersed in a solution containing a lipid, dissolved with a lipid, emulsified with a lipid, mixed with a lipid, combined with a lipid, covalently bonded to a lipid, contained as a suspension in a lipid, contained or complexed with a micelle or liposome, or otherwise associated with a lipid or lipid structure by any means known to those of ordinary skill in the art.
  • the dispersion may or may not result in the formation of liposomes.
  • the actual dosage amount of a composition of the present invention administered to an animal patient can be determined by physical and physiological factors such as body weight, severity of condition, the type of disease being treated, previous or concurrent therapeutic interventions, idiopathy of the patient and on the route of administration.
  • the number of administrations of a preferred dosage and/or an effective amount may vary according to the response of the subject.
  • the practitioner responsible for administration will, in any event, determine the concentration of active ingredient(s) in a composition and appropriate dose(s) for the individual subject.
  • compositions may comprise, for example, at least about 0.1% of an active compound.
  • the an active compound may comprise between about 2% to about 75% of the weight of the unit, or between about 25% to about 60%, for example, and any range derivable therein.
  • the amount of active compound(s) in each therapeutically useful composition may be prepared is such a way that a suitable dosage will be obtained in any given unit dose of the compound. Factors such as solubility, bioavailability, biological half-life, route of administration, product shelf life, as well as other pharmacological considerations will be contemplated by one skilled in the art of preparing such pharmaceutical formulations, and as such, a variety of dosages and treatment regimens may be desirable.
  • a dose may also comprise from about 1
  • microgram/kg/body weight about 5 microgram/kg/body weight, about 10
  • microgram/kg/body weight about 50 microgram/kg/body weight, about 100
  • microgram/kg/body weight about 200 microgram/kg/body weight, about 350
  • microgram/kg/body weight about 500 microgram/kg/body weight, about 1
  • milligram/kg/body weight about 5 milligram/kg/body weight, about 10 milligram/kg/body weight, about 50 milligram/kg/body weight, about 100 milligram/kg/body weight, about 200 milligram/kg/body weight, about 350 milligram/kg/body weight, about 500
  • milligram/kg/body weight to about 1000 mg/kg/body weight or more per administration, and any range derivable therein.
  • a derivable range from the numbers listed herein a range of about 5 mg/kg/body weight to about 100 mg/kg/body weight, about 5 microgram/kg/body weight to about 500 milligram/kg/body weight, etc., can be administered, based on the numbers described above.
  • the one or more compounds of Figs. 7 and 13 are formulated to be administered via an alimentary route.
  • Alimentary routes include all possible routes of administration in which the composition is in direct contact with the alimentary tract.
  • the pharmaceutical compositions disclosed herein may be administered orally, buccally, rectally, or sublingually.
  • these compositions may be formulated with an inert diluent or with an assimilable edible carrier, or they may be enclosed in hard- or soft- shell gelatin capsule, or they may be compressed into tablets, or they may be incorporated directly with the food of the diet.
  • the tablets, troches, pills, capsules and the like may also contain the following: a binder, such as, for example, gum tragacanth, acacia, cornstarch, gelatin or combinations thereof; an excipient, such as, for example, dicalcium phosphate, mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate or combinations thereof; a disintegrating agent, such as, for example, com starch, potato starch, alginic acid or combinations thereof; a lubricant, such as, for example, magnesium stearate; a sweetening agent, such as, for example, sucrose, lactose, saccharin or combinations thereof; a flavoring agent, such as, for example peppermint, oil of wintergreen, cherry flavoring, orange flavoring, etc.
  • a binder such as, for example, gum tragacanth, acacia, cornstarch, gelatin or combinations thereof
  • an excipient such as,
  • the dosage unit form When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier. Various other materials may be present as coatings or to otherwise modify the physical form of the dosage unit. For instance, tablets, pills, or capsules may be coated with shellac, sugar, or both. When the dosage form is a capsule, it may contain, in addition to materials of the above type, carriers such as a liquid carrier. Gelatin capsules, tablets, or pills may be enterically coated. Enteric coatings prevent denaturation of the composition in the stomach or upper bowel where the pH is acidic. See, e.g., U.S. Pat. No. 5,629,001.
  • the basic pH therein dissolves the coating and permits the composition to be released and absorbed by specialized cells, e.g., epithelial enterocytes and Peyer's patch M cells.
  • a syrup of elixir may contain the active compound sucrose as a sweetening agent methyl and propylparabens as preservatives, a dye and flavoring, such as cherry or orange flavor.
  • any material used in preparing any dosage unit form should be pharmaceutically pure and substantially non-toxic in the amounts employed.
  • the active compounds may be incorporated into sustained-release preparation and formulations.
  • compositions of the present disclosure may alternatively be incorporated with one or more excipients in the form of a mouthwash, dentifrice, buccal tablet, oral spray, or sublingual orally- administered formulation.
  • a mouthwash may be prepared incorporating the active ingredient in the required amount in an appropriate solvent, such as a sodium borate solution (Dobell's Solution).
  • the active ingredient may be incorporated into an oral solution such as one containing sodium borate, glycerin and potassium bicarbonate, or dispersed in a dentifrice, or added in a therapeutically - effective amount to a composition that may include water, binders, abrasives, flavoring agents, foaming agents, and humectants.
  • the compositions may be fashioned into a tablet or solution form that may be placed under the tongue or otherwise dissolved in the mouth.
  • suppositories are solid dosage forms of various weights and shapes, usually medicated, for insertion into the rectum. After insertion, suppositories soften, melt or dissolve in the cavity fluids.
  • traditional carriers may include, for example, polyalkylene glycols, triglycerides or combinations thereof.
  • suppositories may be formed from mixtures containing, for example, the active ingredient in the range of about 0.5% to about 10%, and preferably about 1% to about 2%.
  • inducer of expression of PGC-l ⁇ may be administered via a parenteral route.
  • parenteral includes routes that bypass the alimentary tract.
  • the pharmaceutical compositions disclosed herein may be administered for example, but not limited to intravenously, intradermally, intramuscularly, intraarterially, intrathecally, subcutaneous, or intraperitoneally U.S. Pat. Nos. 6,7537,514, 6,613,308, 5,466,468, 5,543,158; 5,641,515; and 5,399,363 (each specifically incorporated herein by reference in its entirety).
  • Solutions of the active compounds as free base or pharmacologically acceptable salts may be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions may also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
  • the pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions (U.S. Patent 5,466,468, specifically incorporated herein by reference in its entirety). In all cases, the form must be sterile and must be fluid to the extent that easy injectability exists.
  • the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (i.e., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and/or vegetable oils.
  • polyol i.e., glycerol, propylene glycol, and liquid polyethylene glycol, and the like
  • suitable mixtures thereof and/or vegetable oils.
  • Proper fluidity may 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 dispersion and by the use of surfactants.
  • the prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.
  • isotonic agents for example, sugars or sodium chloride.
  • Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.
  • aqueous solutions For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose.
  • aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous, and intraperitoneal administration.
  • sterile aqueous media that can be employed will be known to those of skill in the art in light of the present disclosure.
  • one dosage may be dissolved in isotonic NaCl solution and either added hypodermoclysis fluid or injected at the proposed site of infusion, (see for example, "Remington's Pharmaceutical Sciences” l5th Edition, pages 1035-1038 and 1570- 1580).
  • Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with several of the other ingredients enumerated above, as required, followed by filtered sterilization.
  • dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above.
  • the preferred methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
  • a powdered composition is combined with a liquid carrier such as, e.g., water or a saline solution, with or without a stabilizing agent.
  • 7 and 13 may be formulated for administration via various miscellaneous routes, for example, topical (i.e., transdermal) administration, mucosal administration (intranasal, vaginal, etc.) and/or inhalation.
  • topical i.e., transdermal
  • mucosal administration intranasal, vaginal, etc.
  • inhalation inhalation
  • Suitable penetration enhancers include glycerin, alcohols, alkyl methyl sulfoxides, pyrrolidones and luarocapram.
  • Possible bases for compositions for topical application include polyethylene glycol, lanolin, cold cream and petrolatum as well as any other suitable absorption, emulsion or water-soluble ointment base.
  • Topical preparations may also include emulsifiers, gelling agents, and antimicrobial preservatives as necessary to preserve the active ingredient and provide for a homogenous mixture.
  • Transdermal administration of the present invention may also comprise the use of a "patch".
  • the patch may supply one or more active substances at a predetermined rate and in a continuous manner over a fixed period of time.
  • the pharmaceutical compositions may be delivered by eye drops, intranasal sprays, inhalation, and/or other aerosol delivery vehicles.
  • Methods for delivering compositions directly to the lungs via nasal aerosol sprays has been described e.g., in U.S. Pat. Nos. 5,756,353 and 5,804,212 (each specifically incorporated herein by reference in its entirety).
  • the delivery of drugs using intranasal microparticle resins are examples of drugs using intranasal microparticle resins
  • aerosol refers to a colloidal system of finely divided solid of liquid particles dispersed in a liquefied or pressurized gas propellant.
  • the typical aerosol of the present invention for inhalation will consist of a suspension of active ingredients in liquid propellant or a mixture of liquid propellant and a suitable solvent.
  • Suitable propellants include hydrocarbons and hydrocarbon ethers.
  • Suitable containers will vary according to the pressure requirements of the propellant.
  • Administration of the aerosol will vary according to subject’s age, weight and the severity and response of the symptoms.
  • kits may comprise a suitably aliquoted the one or more compounds of Figs. 7 and 13 and, in some cases, one or more additional agents.
  • the component(s) of the kits may be packaged either in aqueous media or in lyophilized form.
  • the container means of the kits will generally include at least one vial, test tube, flask, bottle, syringe or other container means, into which a component may be placed, and preferably, suitably aliquoted. Where there are more than one component in the kit, the kit also will generally contain a second, third or other additional container into which the additional components may be separately placed.
  • kits of the present invention also will typically include a means for containing the one or more compounds of Figs. 7 and 13 and any other reagent containers in close confinement for commercial sale.
  • Such containers may include injection or blow-molded plastic containers into which the desired vials are retained.
  • the liquid solution is an aqueous solution, with a sterile aqueous solution being particularly preferred.
  • the one or more compounds of Figs. 7 and 13 composition(s) may be formulated into a syringeable composition.
  • the container means may itself be a syringe, pipette, and/or other such like apparatus, from which the formulation may be applied to an infected area of the body, injected into an animal, and/or even applied to and/or mixed with the other components of the kit.
  • the components of the kit may be provided as dried powder(s). When reagents and/or components are provided as a dry powder, the powder can be reconstituted by the addition of a suitable solvent. It is envisioned that the solvent may also be provided in another container means.
  • Quantitative high-throughput screening A titration-based approach that efficiently identifies biological activities in large chemical libraries. Proc. Natl. Acad. Sci. 103,
  • Quantitative high-throughput screening a titration-based approach that efficiently identifies biological activities in large chemical libraries. Proc. Natl. Acad. Sci. U. S. A. 103, 11473-8. doi: 10.1073/pnas.0604348103.
  • Phenotypic Drug Discovery Is a Viable, Neoclassic Pharma Strategy. J. Med. Chem. 55, 4527-4538. doi: l0. l02l/jm20l649s.
  • NfsA-like nitroreductases from Neisseria meningitidis and Bartonella henselae for enzyme-prodrug therapy, targeted cellular ablation, and dinitrotoluene bioremediation. Biotechnol. Lett. 40, 359-367. doi: l0. l007/sl0529-0l7-2472-5.
  • ablation facilitates cell-specific regenerative studies in zebrafish. Methods 62, 232-240. doi: l0. l0l6/j.ymeth.20l3.03.0l7.
  • reticulum stress induced by tunicamycin and thapsigargin protects against transient ischemic brain injury.

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Abstract

La présente invention concerne des compositions et des méthodes de traitement ou de prévention de la cécité chez un sujet. La méthode comprend une étape d'administration à un sujet ayant une maladie résultant en la mort des photorécepteurs, la mort des cellules rétiniennes, ou leurs combinaisons, d'un ou de plusieurs composés de la présente invention.
PCT/US2019/029258 2018-04-27 2019-04-26 Médicaments favorisant la survie des photorécepteurs du bâtonnet rétinien Ceased WO2020036658A2 (fr)

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DE19718826A1 (de) * 1997-05-05 1998-11-12 Marion S Dr Eckmiller Verwendung biologisch aktiver Wirkstoffe zum Beeinflussen des Extrazellulär-Raumes von Sinneszellen und Verfahren zur Wirkstoff-Administrationssteuerung
WO2007150046A2 (fr) * 2006-06-22 2007-12-27 Sirion Therapeutics, Inc Procédés et compositions pour traiter des conditions ophtalmiques par la modulation de l'activité de mégaline
ITMI20090284A1 (it) * 2009-02-26 2010-08-27 Consiglio Nazionale Ricerche Uso di inibitori della serina palmitoiltrasferasi per la prevenzione e il rallentamento delle degenerazioni retiniche ereditarie e relative composizioni
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