[go: up one dir, main page]

WO2015073527A1 - Nitroalcène-tocophérols et analogues de ceux-ci utilisables dans le cadre du traitement et de la prévention d'affections associées à l'inflammation - Google Patents

Nitroalcène-tocophérols et analogues de ceux-ci utilisables dans le cadre du traitement et de la prévention d'affections associées à l'inflammation Download PDF

Info

Publication number
WO2015073527A1
WO2015073527A1 PCT/US2014/065203 US2014065203W WO2015073527A1 WO 2015073527 A1 WO2015073527 A1 WO 2015073527A1 US 2014065203 W US2014065203 W US 2014065203W WO 2015073527 A1 WO2015073527 A1 WO 2015073527A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
nitroalkene
formula
pharmaceutical composition
pharmaceutically
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/US2014/065203
Other languages
English (en)
Inventor
Carlos Ignacio BATTHYANY DIGHIERO
Gloria Virginia LOPEZ GONZALEZ
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.)
Complexa Inc
Original Assignee
Complexa Inc
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 Complexa Inc filed Critical Complexa Inc
Publication of WO2015073527A1 publication Critical patent/WO2015073527A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
    • C07D311/70—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with two hydrocarbon radicals attached in position 2 and elements other than carbon and hydrogen in position 6
    • C07D311/72—3,4-Dihydro derivatives having in position 2 at least one methyl radical and in position 6 one oxygen atom, e.g. tocopherols
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
    • C07D311/66—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached in position 2
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/70—Mechanisms involved in disease identification
    • G01N2800/7095—Inflammation

Definitions

  • organic acids include formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, mesylic, salicylic, 4-hydrobenzoic, phylacetic, mandelic, embonic, methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, 2- hydroxyethanesulfonic, toluenesulfonic, sulfanilic, cyclohyexylaminosuflonic, stearic, algenic, ⁇ - hydrobutyric, galactaric and galacturnoic acid.
  • Suitable pharmaceutically-acceptable base addition salts of compounds of Formula I, Formula II, and Formula III include metallic salts, such as salts made from aluminum, calcium, lithium, magnesium, potassium, sodium and zinc, or salts made from organic bases including primary, secondary and tertiary amines, substituted amines including cyclic amines, such as caffeine, arginine, diethylamine, N-ethyl piperidine, histidine, glucamine, isopropylamine, lysine, morpholine, N-ethyl morpholine, piperazine, triethylamine, trimethylamine.
  • metallic salts such as salts made from aluminum, calcium, lithium, magnesium, potassium, sodium and zinc
  • organic bases including primary, secondary and tertiary amines, substituted amines including cyclic amines, such as caffeine, arginine, diethylamine, N-ethyl piperidine, histidine, glucamine, isopropylamine
  • powders, pills, capsules and tablets can employ solid excipients such as starches, sugars, kaolin, lubricants, binders, disintegrating agents, antioxidants and the like.
  • Parenteral compositions typically employ sterile water as a carrier and optionally other ingredients, such as solubility aids.
  • injectable solutions can be prepared, for example, using a carrier comprising a saline solution, a glucose solution or a solution containing a mixture of saline and glucose. Further guidance for methods suitable for use in preparing pharmaceutical compositions is provided in Remington: The Science and Practice of Pharmacy, 21 st edition (Lippincott Williams & Wilkins, 2006).
  • composition comprising an effective amount of pharmaceutically-acceptable salts of active agent and a pharmaceutically-acceptable carrier.
  • the active agent may be mixed with one or more stabilizers such as, for example, antioxidants, vitamin E, vitamin C, ⁇ -carotene, wheat germ oil and the like, and in some embodiments the capsule may be combined with one or more solubilizers such as, for example, surfactants, hydrophilic or hydrophobic solvents, oils, or combinations thereof.
  • stabilizers such as, for example, antioxidants, vitamin E, vitamin C, ⁇ -carotene, wheat germ oil and the like
  • solubilizers such as, for example, surfactants, hydrophilic or hydrophobic solvents, oils, or combinations thereof.
  • propylene glycol esters may include, for example, propylene carbonate, propylene glycol monoacetate, propylene glycol diacetate, propylene glycol fatty acid esters, acetylated propylene glycol fatty acid esters, and mixtures thereof.
  • propylene glycol fatty acid esters may be a propylene glycol fatty acid monoester, propylene glycol fatty acid diester, or mixture thereof.
  • propylene glycol ester may be propylene glycol monocaprylate, propylene glycol dicaprylate, propylene glycol dicaprate, propylene glycol dicaprylate/dicaprate, and mixtures thereof.
  • the coatings of various embodiments may further include one or more film forming materials and/or binders and/or other conventional additives such as lubricants, fillers, antiadherents, antioxidants, buffers, solubilizers, dyes, chelating agents, disintegrants, and/or absorption enhancers.
  • lubricants such as lubricants, fillers, antiadherents, antioxidants, buffers, solubilizers, dyes, chelating agents, disintegrants, and/or absorption enhancers.
  • Surfactants may act as both solubilizers and absorption enhancers.
  • Capsular materials may further include one or more preservatives, coloring and opacifying agents, flavorings and sweeteners, sugars, gastroresistant substances, or combinations thereof.
  • Suitable preservative and colorant are known in the art and include, for example, benzoic acid, para-oxybenzoate, caramel colorant, gardenia colorant, carotene colorant, tar colorant, and the like.
  • one or more flavoring agents may be included the contents of the core of the gelatin capsule or in one or more coating layers of the capsule, or a combination thereof.
  • the gel capsules of embodiments may include at least one coating layer including one or more secondary agent.
  • a layer including one or more secondary agent may be of sufficient thickness to prevent oxidative degradation of the one or more secondary agent.
  • the thickness of this layer may be from about 5 to about 400 microns, about 10 to about 200 microns, about 20 to about 100 microns, or in certain embodiments, from about 40 to about 80 microns.
  • the thickness of such layers may be expressed in terms of percentage weight gain based on the total weight of the capsule.
  • a layer including one or more secondary agents may create a weight gain of about 0.05 to about 20 %, about 0.1 to about 10%, about 0.1 to about 5%, and in particular embodiments about 0.25 to about 1 %.
  • a coating layer containing one or more secondary agent may further include at least one compound to prevent oxidative degradation.
  • at least one polymer such as, but not limited to cellulose derivatives such as hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinylpyrrolidone, polyvinylpyrrolidone/vinyl acetate copolymer, ethyl cellulose aqueous dispersions, and combinations thereof, preferably
  • the secondary agent may be provided as a homogenous coating solution or a heterologous suspension in a pharmaceutically acceptable solvent.
  • a pharmaceutically acceptable solvent may be an aqueous or organic solvent such as, for example, methanol, ethanol, isopropranol, ethylene glycol, acetone, or mixtures thereof.
  • pharmaceutically acceptable solvents may include, but are not limited to, polypropylene glycol, polypropylene glycol, polyethylene glycol, for example, polyethylene glycol 600, polyethylene glycol 900, polyethylene glycol 540, polyethylene glycol 1450, polyethylene glycol 6000, polyethylene glycol 8000, and the like; pharmaceutically acceptable alcohols that are liquids at about room temperature, for example, propylene glycol, ethanol, 2-(2-ethoxyethoxy)ethanol, benzyl alcohol, glycerol, polyethylene glycol 200, polyethylene glycol 300, polyethylene glycol 400 and the like, polyoxyethylene castor oil derivatives, for example, polyoxyethyleneglycerol triricinoleate or polyoxyl 35 castor oil, polyoxyethyleneglycerol oxystearate, RH 40 (poly ethylenegly col 40 hydrogenated castor oil) or RH 60 (polyethyleneglycol 60 hydrogenated castor oil), and the like, saturated polyglycolized glycerides
  • capsules may be produced by a method including the steps of preparing a sheet of an outer coating layer and one or more sheets of other layers, laminating the sheets, drying the laminated sheets to obtain a dried sheet, and encapsulating the active agent or the compounds and one or more secondary agents within the dried sheet on a rotary filler to form a seamed capsule.
  • seamless capsules may be produced using an instrument equipped with two or more nozzles arranged concentrically.
  • gelatin capsules may be manufactured as, for example, a two-piece, sealed or unsealed hard gelatin capsule.
  • a gelatin capsule including the active agent may be formed by the encapsulation of a dose of the active agent in a gelatin capsule.
  • the gelatin capsule may be made of, for example, gelatin, glycerol, water, a flavoring, a coloring agent and combinations thereof, and the compound dosage may be, for example, 0.001 to 1000 milligrams.
  • One compound dosage range is 0.01 to 500 mg.
  • Preferred compound dosages include 1, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, and 750 mg of the active ingredient.
  • the manufacturing process of such embodiments may include the steps of combining gelswatch ingredients, melting and forming a liquefied gelswatch, delivering the liquefied gelswatch and the active agent to an encapsulation machine, such as the Capsule filler - Zanasi 70C from IMA Pharma (Tonawanda, NY), encapsulating a dose of the compound drying the encapsulated dose, washing the encapsulated dose and packaging the capsules for shipment.
  • an encapsulation machine such as the Capsule filler - Zanasi 70C from IMA Pharma (Tonawanda, NY
  • the gelswatch ingredients may include any ingredients described herein that are useful in the production of gelatin capsules such as, for example, gelatin or a gelatin substitute such as modified starch or other suitable gelatin substitute known in the art, a softener such as glycerol or sorbitol or other suitable polyol or other gelatin softener known in the art, a flavoring agent such as strawberry flavor Firmenich #52311 A or other suitable gelatin capsule flavoring known in the art and optionally a coloring agent such as keratin or other suitable gelatin capsule coloring agent known in the art.
  • gelatin or a gelatin substitute such as modified starch or other suitable gelatin substitute known in the art
  • a softener such as glycerol or sorbitol or other suitable polyol or other gelatin softener known in the art
  • a flavoring agent such as strawberry flavor Firmenich #52311 A or other suitable gelatin capsule flavoring known in the art
  • optionally a coloring agent such as keratin or other suitable gelatin
  • the gel capsule may be formed from a gelswatch mixture of about 45 parts by weight of gelatin, about 20 parts by weight of glycerol, about 35 parts by weight of water and about 0.5 or more parts by weight of flavoring.
  • the gelswatch ingredients may be heated to about 60° C to 70° C and mixed together to form liquefied gelswatch.
  • the liquefied gelswatch and the active agent may then be poured into an encapsulation machine.
  • the encapsulation machine such as the Capsule filler - Zanasi 70C from IMA Pharma (Tonawanda, NY), then forms the capsule by encapsulating the dose of the compound into a gelatin capsule.
  • the one or more coatings on the capsule may be applied by any technique known in the art including, but not limited to, pan coating, fluid bed coating or spray coating, and the one or more coatings may be applied, for example, as a solution, suspension, spray, dust or powder.
  • a polymeric coating may be applied as aqueous-based solutions, organic-based solutions or dispersions containing and, in some embodiments, one or more secondary agent.
  • polymer-containing droplets may atomized with air or an inert gas and sprayed onto the a core containing the active agent, and in some embodiments, heated air or inert gas may be added to facilitate evaporation of the solvent and film formation.
  • the processing parameters of spray rate and bed temperature must be controlled to limit solubilization and capsule
  • agglomeration a high bed temperature may result in evaporation of residual water from the capsule shell, causing the capsule to become brittle.
  • coating uniformity which includes mass variance of the coated capsules and variance of the content of the coated active agent and accuracy of deposition must be evaluated.
  • Gel capsules of various embodiments of the invention may be of any shape such as, but not limited to, round, oval, tubular, oblong, twist off, or a non-standard shape (e.g., animal, tree, star, heart, etc.), and the size of the capsule may vary in accordance to the volume of the fill composition intended to be contained therein.
  • hard or soft gelatin capsules may be manufactured using conventional methods as a single body unit comprising the standard capsule shape.
  • hard gel capsules may be manufactured using conventional methods in standard shapes and various standard sizes, such as those designated (000), (00), (0), (1), (2), (3), (4), and (5) where the largest number corresponds to the smallest size.
  • Nonstandard shapes may be used as well.
  • Suitable vegetable oils including sesame, castor, corn, and cottonseed oils, include those listed in R. C. Rowe and P. J. Shesky, Handbook of Pharmaceutical Excipients, (2006), 5th ed., which is incorporated herein by reference in its entirety.
  • Suitable polyethoxylated vegetable oils include but are not limited to, CremaphorTM EL or RH series (available from BASF), EmulphorTM EL-719 (available from Stepan products), and EmulphorTM EL-620P (available from GAF).
  • Suitable polyethoxylated castor oils include, but are not limited to, the NikkolTM HCO series (available from Nikko Chemicals Co. Ltd.), such as Nikkol HCO-30, HC-40, HC-50, and HC-60 (polyethylene glycol-30 hydrogenated castor oil, polyethylene glycol-40 hydrogenated castor oil, polyethylene glycol-50 hydrogenated castor oil, and polyethylene glycol-60 hydrogenated castor oil, EmulphorTM EL-719 (castor oil 40 mole- ethoxylate, available from Stepan Products), the CremophoreTM series (available from BASF), which includes Cremophore RH40, RH60, and EL35 (polyethylene glycol-40 hydrogenated castor oil, polyethylene glycol-60 hydrogenated castor oil, and polyethylene glycol-35 hydrogenated castor oil, respectively), and the Emulgin® RO and HRE series (available from Cognis PharmaLine).
  • Other suitable polyoxy ethylene castor oil derivatives include those listed in R. C. Rowe and P
  • Sterol As used herein, the term “sterol” refers to a compound, or mixture of
  • PEG-5 soya sterol, NikkolTM BPS-5 available from Nikko
  • PEG- 10 soya sterol, NikkolTM BPS-10 available from Nikko
  • PEG-20 soya sterol, NikkolTM BPS-20 available from Nikko
  • PEG-30 soya sterol, NikkolTM BPS-30 available from Nikko
  • the average molecular weight of the polyethylene glycol is from about 200 to about 9000. In some embodiments, the average molecular weight of the polyethylene glycol is from about 200 to about 5000. In some embodiments, the average molecular weight of the polyethylene glycol is from about 200 to about 900. In some embodiments, the average molecular weight of the polyethylene glycol is about 400.
  • Suitable polyethylene glycols include, but are not limited to polyethylene glycol-200, polyethylene glycol-300, polyethylene glycol-400, polyethylene glycol-600, and polyethylene glycol-900. The number following the dash in the name refers to the average molecular weight of the polymer. In some embodiments, the polyethylene glycol is polyethylene glycol-400.
  • Suitable propylene glycol fatty acid esters include, but are not limited to, propylene glycol laurates: LauroglycolTM FCC and 90 (available from Gattefosse); propylene glycol caprylates: CapryolTM PGMC and 90 (available from Gatefosse); and propylene glycol dicaprylocaprates: LabrafacTM PG (available from Gatefosse).
  • Stearoyl macrogol glyceride refers to a polyglycolized glyceride synthesized predominately from stearic acid or from compounds derived
  • hydroxyethylcellulose methylhydroxyethylcellulose, starch, sodium starch glycolate, pregelatinized starch, a calcium phosphate, a metal carbonate, a metal oxide, or a metal aluminosilicate.
  • Exemplary excipients or carriers for use in solid and/or liquid dosage forms include, but are not limited to:
  • Liponic 70-NC and 76-NC available from Lipo Chemical
  • Neosorb available from Roquette
  • Partech SI available from Merck
  • Sorbogem available from SPI Polyols
  • Starch, sodium starch glycolate, and pregelatinized starch include, but are not limited to, those described in R. C. Rowe and P. J. Shesky, Handbook of Pharmaceutical Excipients,
  • sphingomyelin 34 mM in chloroform:methanol (1 : 1 v/v). Organic solvents were evaporated by centrifugation under vacuum (Speed Vac) for 1 hr at 30°C. Lipids were resuspended in 1 ml of Tris 20mM, Mes lOmM, NaCl 120mM, DTP A ⁇ and acetic acid lOmM buffer at 50°C for 20 min. Suspension was sonicated at least 10 times at 10% max frequency and till solution became clear. Small Unilamellar Vesicles (SUVs) with NATOH were incubated with b- Mercaptoethanol 250 ⁇ from SIGMA (St. Louis, MO). UV -visible spectrums were taken every min up to 10 min. Spectrums were acquired in a Varian Cary 50 Bio from Agilent Technologies (Santa Clara, CA).
  • Reactive scheme 2 prepares a trolox derivative with a nitroalkene at position 5 of the chroman ring and an unprotected hydroxyl group at position 6 of the chroman ring.
  • scheme 4 shows that other electrophilic centers may be introduced at position 5.
  • nitroalkenes groups are incorporated at the phytenyl tail of tocotrienols as shown in scheme 5.
  • Scheme 5 includes not only single substitutions of nitro groups on the phytenyl chain but also multiple substitutions of nitro groups by increasing the AgN0 2 to tocotrienol mole ratio of the reaction.
  • step 1 To the solution of step 1 (1.24 g, 2.24 mmol) in dry acetonitrile (20 mL), NMMO (1.05 g, 9.0 mmol, 4 equiv) was added. After stirring overnight at room temperature, the solvent was evaporated and the crude residue purified by column chromatography (Hex/EtOAc 15:1), affording 3 (1.01 g, 93%) as yellow dense oil.
  • step 2 The solution of step 2 (0.150 g, 0.3 mmol) was added in a mixture of 1.2 mL CH3N02 and equivalent amount of CH3COONH4. The mixture was stirred at 100°C for 2 h. The solvent is then evaporated and H20 and diethyl ether were added. The organic layer was washed with H20 (2x 50 mL), HC1 3N (2x 25 mL), and saturated aqueous NaCl, dried, and the solvent was evaporated. The crude residue purified by column chromatography (Hex/EtOAc 9:1), affording NATOH (0.04 g, 27%) as yellow dense oil.
  • Step 1 Preparation of 6-hydroxy-2,5,7,8-tetramethylchroman carboxylic acid methyl ester (4).
  • Step 2 Preparation of 6-acetoxy-5-bromomethyl-2,7,8-trimethylchroman carboxylic acid methyl ester.
  • a round-bottom flask was charged with 1.58 g (0.006 mol) of the solution of step 1, 33.0 mL of dichloromethane. The mixture was stirred at room temperature in the dark, and a solution of 0.3 mL (0.006 mol) of bromine in 4.5 mL of dichloromethane was added dropwise. Stirring continued at rt for 2 h after completion of the bromine addition; the resulting solution was dark, but no bromine color or vapor was detectable. The mixture was purged with a stream of nitrogen to remove most of the HBr present, then stripped to dryness under reduce pressure.
  • the crude intermediate product was dissolved with 13 mL of dichloromethane, and treated with 1 1.0 mL of glacial acetic acid, 3.0 mL of acetic anhydride, and 1 drop of cone, sulfuric acid. After stirring overnight at rt, the mixture was treated with 60.0 mL of water and stirred for 1 h. The mixture was transferred to a separatory funnel and the layers were separated. The aqueous layer was extracted with dichloromethane. The combined organic layers were washed with brine, dried (MgS04), and solvent evaporated under reduced pressure. The crude residue purified by column chromatography (Hex/Et20, 7:3), affording 5 (2.0 g, 87% yield) as white solid.
  • Step 3 Preparation of 6-acetoxy-5-formyl-2,7,8-trimethylchroman carboxylic acid methyl ester.
  • Step 4 Preparation (E)-6-hydroxy-2,7,8-trimethyl-5-(2-nitrovinyl)chromancarboxylic acid methyl ester (NATxME).
  • step 3 The solution of step 3 (0.18 g, 0.56 mmol) was added in a mixture of 2.5 mL anhyd CH3N02 and equivalent amount of CH3COONH4. The mixture was stirred at 100 oC for 2 h. The solvent is then evaporated and H20 and diethyl ether were added. The organic layer was washed with H20 (2 ⁇ 50 mL), HC1 3 N (2 ⁇ 25 mL), and satd aqueous NaCl, dried, and the solvent was evaporated. The crude residue purified by column chromatography (Hex/Et20, 7:3), affording NATxME (0.032 g, 18%) as yellow dense oil.
  • FIG. 1 a Western Blot analysis was performed to analyze HO-1 protein expression after exposure to the NA-Trolox-ME compound.
  • Raw 264.7 cells a murine macrophage cell line catalog no. TIB-71TM from ATCC (Manassas, VA), were treated with NA-Trolox-ME during 18 hs and after that time cells were lysed.
  • Total protein concentration was measured with Pierce BCA assay from Thermo Fisher Scientific (Rockford, IL).
  • For electrophoresis 30 ug of protein was used in each line. The proteins were electrophoresed on a Tris/glycine SDS- polyacrylamide gradient gel (10-15%) and transferred to nitrocellulose membrane.
  • FIG. 2 a Western Blot analysis was performed to analyze GCLM protein expression after exposure to the NA-Trolox-ME compound.
  • Raw 264.7 cells a murine macrophage cell line catalog no. TIB-71TM from ATCC (Manassas, VA), were treated with NA-Trolox-ME for 18 hs were lysed, and total protein concentration was measured with Pierce BCA assay from Thermo Fisher Scientific (Rockford, IL). For electrophoresis, 30 ug of protein was used in each line. The proteins were electrophoresed on a Tris/glycine SDS-polyacrylamide gradient gel (10-15%) and transferred to nitrocellulose membrane.
  • the primary antibodies used for detection was rabbit polyclonal anti-GCLM from Abeam (Cambridge, MA). Blots were visualized using horseradish peroxidase-conjugated secondary antibodies and ECL Plus detection system from GEHealthcare (Little Chalfont, UK). Blots were detected with a scanner. Protein expression was quantified with ImageQuant TL7.0 software from GE Healthcare (Little Chalfont, UK).
  • FIG. 1 and FIG. 2 illustrate that the nitroalkene tocopherol analog NA-Trolox-ME performs at least similar to electrophilic fatty acid derivatives (positive control OA-NO 2 ) that mediate cytoprotective cell signaling reactions via phase 2 gene expression.
  • Q RT-PCR Quantitative Reverse Transcription Polymerase Chain Reaction analysis
  • Raw 264.7 cells a murine macrophage cell line catalog no. TIB- 71TM from ATCC (Manassas, VA), were treated with NA-Trolox-ME for 5 hours.
  • RNA extraction was performed using TRIzol ® from Invitrogen Life Technologies (Carlsbad, CA) reagent according to the manufacturer's instructions. lOOng/ml of RNA was retrotranscripted using the iScriptTM cDNA kit from Bio-Rad (Hercules, CA). Samples were analyzed with TaqMan fast universal PCR master mixture using HO-1, GCLM, NQO-1 and GAPDH primers.
  • Figure 3 exemplifies the dose dependence of HO-1 protein expression buy the cells.
  • FIGS. 4-6 enzyme-linked immunosorbent assays (ELISA) were performed on raw cells to quantify the concentration of cytokines after exposure to NA-Trolox-ME.
  • Raw 264.7 cells activated with LPS (50 ng/ml) were treated with NA-Trolox-ME. for 18 hours.
  • Cell medium was taken and the concentration of cytokines (TNF-a, MCP-1 & IL-6) was determined using ELISA kits according to the manufacturer's instructions (R & D systems, Minneapolis, MN).
  • NA-Tx- TOH is similar to nitro-fatty acid derivatives that potently inhibit stimulus-induced proinflammatory cytokine release from a variety of cells, highlighting the diversity of mechanisms by which nitroalkenes mediate signal transduction. Similar to the endogenous nitroalkenes the NA-Tx-TOH inhibited LPS-induced cytokine release in the macrophage cell line THP-1, with significant reductions observed in interleukin-6 (IL-6), tumor necrosis factor alpha (TNFa), and monocyte chemotractant protein- 1 (MCP-1).
  • IL-6 interleukin-6
  • TNFa tumor necrosis factor alpha
  • MCP-1 monocyte chemotractant protein- 1
  • FIGS. 7-9 In vivo experiments in mice were performed to detect and quantify mR A transcription levels for HO-1, GCLM, and NQOl after administration of NA-a-TOH.
  • C57BL 6J Mice were treated with NATOH or NA-Trolox-ME (50mg/kg) via intraperitoneal injection or gavage. After 5 hs, mice were sacrificed and organ samples were taken in order to perform Q RT-PCR in vivo analysis of Nrf2/Keapl reporters (HO-1, GCLM & NQOl).
  • Total RNA was extracted with TRIzol ® Invitrogen Life Technologies (Carlsbad, CA) according to
  • FIGS. 7-9 illustrate the over expression of HO-1, GCLM and NQOl as compared to the positive control groups.
  • RP-LC-MS reverse phase liquid chromatography mass spectroscopy
  • Another model includes in vivo incorporation of NATOH into LDL by treating mice with NATOH at a 50mg/kg dose via intraperitoneal injection or gavage. After three days of consecutive administration of the compound, LDL was purified by sequential ultracentrifuation in KBr gradients to show selective secretion of the NATOH into circulation via the VLDL-LDL system by the Alpha-Tocopherol Transfer Protein (a-TTP) after hepatic uptake.
  • a-TTP Alpha-Tocopherol Transfer Protein
  • Other animal models include: (a) atherosclerosis treatment models using rLDL -/- knockout mice and Apo E -/- knockout mice; (b) systemic inflammation or sepsis models using Ccl20 reporter and cecal ligation and perforation; (c) ischemia-reperfusion models characterized by myocardial infarction by isoproterenol and ischemia-reperfusion injury in rat kidneys; and (d) ventilator induced lung injury in rats.
  • the Ccll20 reporter mouse model is a transgenic mouse harboring the transcriptional fusion Ccl20-luciferase as a reporter of pro-inflammatory response.
  • NA-a-TOH 100 uM
  • b-Mercaptoethanol lOmM, SIGMA
  • FIG. 11 the mixture at the end of the reaction described in association with FIG. 10 was analyzed by RP-HPLC (C-18, 2.1 mm ID x 150 mm, 5um, VYDAC from Grace Davison Discovery Sciences (Deerfield, IL)). Column was equilibrated in 50% Acetonitrile from
  • Compounds of the invention would be useful to treat arthritis, including but not limited to rheumatoid arthritis, spondyloarthropathies, gouty arthritis, osteoarthritis, systemic lupus erythematosus and juvenile arthritis. Such compounds of the invention would also be useful in the treatment of asthma, bronchitis, menstrual cramps, preterm labor, tendinitis, bursitis, liver disease including hepatitis, skin related conditions such as psoriasis, eczema, burns and dermatitis, and from post-operative inflammation including from ophthalmic surgery such as cataract surgery and refractive surgery.
  • the method above would be useful for, but not limited to, treating and preventing inflammation-related cardiovascular disorders in a subject.
  • the method would be useful for treatment and prevention of vascular diseases, coronary artery disease, aneurysm,
  • the compounds would be useful for, but not limited to, the treatment of angiogenesis-related disorders in a subject.
  • the compounds are administered to a subject in need of angiogenesis inhibition.
  • Treatment may include combination therapies comprising a compound of the present invention with one or more secondary agents including but not limited to the following: statins, beta-blockers, calcium antagonists, angiotensin-converting enzyme inhibitors, inhibitors of angiotensin II receptors, diuretics, and anti-inflammatory agents such as but not limited to acetyl salicylic acid and omega- 3 fatty acids.
  • secondary agents including but not limited to the following: statins, beta-blockers, calcium antagonists, angiotensin-converting enzyme inhibitors, inhibitors of angiotensin II receptors, diuretics, and anti-inflammatory agents such as but not limited to acetyl salicylic acid and omega- 3 fatty acids.
  • Nrf2 gene transfer induces antioxidant enzymes and suppresses smooth muscle cell growth in vitro and reduces oxidative stress in rabbit aorta in vivo, Arterioscler Thromb Vase Biol 27, 741- 747.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Pain & Pain Management (AREA)
  • Rheumatology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne une nouvelle stratégie pharmaceutique de traitement de l'athérosclérose fondée sur le rôle pathologique principal joué par les lipoprotéines de basse densité et l'inflammation chronique dans l'athérogénèse. Cette approche pharmacologique implique que le composé hybride soit sélectivement incorporé dans les particules de lipoprotéines durant le métabolisme normal du fait de la présence de la structure chromanol du tocophérol. Une fois que le composé hybride nitroalcène-analogue de vitamine E est incorporé dans la LDL, cette dernière le transportera dans tout l'organisme et, notamment, dans les lésions athéroscléreuses, où le composé hybride pourra exercer ses efficaces propriétés anti-inflammatoires et anti-athérogènes similaires à celles des acides gras nitrés, mais sans les inconvénients de la β-oxydation, ni le manque de contrôle sur le ciblage et la localisation du composé.
PCT/US2014/065203 2013-11-12 2014-11-12 Nitroalcène-tocophérols et analogues de ceux-ci utilisables dans le cadre du traitement et de la prévention d'affections associées à l'inflammation Ceased WO2015073527A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201361903068P 2013-11-12 2013-11-12
US61/903,068 2013-11-12

Publications (1)

Publication Number Publication Date
WO2015073527A1 true WO2015073527A1 (fr) 2015-05-21

Family

ID=53057961

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2014/065203 Ceased WO2015073527A1 (fr) 2013-11-12 2014-11-12 Nitroalcène-tocophérols et analogues de ceux-ci utilisables dans le cadre du traitement et de la prévention d'affections associées à l'inflammation

Country Status (1)

Country Link
WO (1) WO2015073527A1 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018037279A1 (fr) 2016-08-23 2018-03-01 Institut Pasteur De Montevideo Dérivés de nitroalcène trolox et leurs méthodes d'utilisation dans le traitement et la prévention d'états associés à une inflammation
US10537541B2 (en) 2015-10-02 2020-01-21 Complexa Inc. Treatment of focal segmental glomerular sclerosis (FSGS) using therapeutically effective oral doses of 10-nitro-9(E)-octadec-9-enoic acid
WO2020058917A1 (fr) * 2018-09-20 2020-03-26 Institut Pasteur De Montevideo Procédés de traitement d'états associés à une inflammation à l'aide de modulateurs anti-inflammatoires et métaboliques pluripotents
US11400066B2 (en) 2016-10-14 2022-08-02 Institut Pasteur De Montevideo Methods of treatment of inflammation related conditions using pluripotent anti-inflammatory and metabolic modulators
US11608342B2 (en) 2015-07-07 2023-03-21 H. Lundbeck A/S PDE9 inhibitors with imidazo triazinone backbone and imidazo pyrazinone backbone for treatment of peripheral diseases
US12006319B2 (en) 2018-05-25 2024-06-11 Cardurion Pharmaceuticals, Inc. Monohydrate and crystalline forms of 6-[(3S,4S)-4-methyl-1-(pyrimidin-2-ylmethyl)pyrrolidin-3-yl]-3-tetrahydropyran-4-yl-7H-imidazo[1,5-a]pyrazin-8-one
US12213975B2 (en) 2018-08-31 2025-02-04 Cardurion Pharmaceuticals, Inc. PDE9 inhibitors for treating sickle cell disease
US12280029B2 (en) 2016-10-14 2025-04-22 Institut Pasteur De Montevideo Methods of treatment of inflammation related conditions using pluripotent anti-inflammatory and metabolic modulators

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010129763A1 (fr) * 2009-05-08 2010-11-11 Perricone Nicholas V Compositions à base de nitroalcène

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010129763A1 (fr) * 2009-05-08 2010-11-11 Perricone Nicholas V Compositions à base de nitroalcène

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CHEN, W. ET AL.: "Synthesis and screening of novel vitamin E derivatives for anticancer functions.", EUR. J. MED. CHEM., vol. 58, 2012, pages 72 - 83 *
LAMAS BERVEJILLO, M. ET AL.: "Estudio del potencial anti-asterogenico del AAN02 in vivo.", TESINA DEL GRADO DE LA LICENCIATURA EN BIOQUIMICA. FACULTAD DE CIENCIAS, UDELAR., February 2012 (2012-02-01), pages 5 - 6 *
LOPEZ, G. V. ET AL.: "Second generation of a-tocopherol analogs-nitric oxide donors: Synthesis, physicochemical, and biological characterization.", BIOORG. MED. CHEM., vol. 15, 2007, pages 6262 - 6272, XP022169173, DOI: doi:10.1016/j.bmc.2007.06.019 *
SETIADI, D. H. ET AL.: "Vitamin E models. Conformational analysis and stereochemistry of tetralin, chroman, thiochroman and selenochroman.", J. MOL. STRUCT (THEOCHEM)., vol. 594, 2002, pages 161 - 172 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11608342B2 (en) 2015-07-07 2023-03-21 H. Lundbeck A/S PDE9 inhibitors with imidazo triazinone backbone and imidazo pyrazinone backbone for treatment of peripheral diseases
US10537541B2 (en) 2015-10-02 2020-01-21 Complexa Inc. Treatment of focal segmental glomerular sclerosis (FSGS) using therapeutically effective oral doses of 10-nitro-9(E)-octadec-9-enoic acid
WO2018037279A1 (fr) 2016-08-23 2018-03-01 Institut Pasteur De Montevideo Dérivés de nitroalcène trolox et leurs méthodes d'utilisation dans le traitement et la prévention d'états associés à une inflammation
US20180057475A1 (en) * 2016-08-23 2018-03-01 Institut Pasteur De Montevideo Nitroalkene Trolox Derivatives and Methods of Use Thereof In The Treatment And Prevention of Inflammation Related Conditions
US9994541B2 (en) * 2016-08-23 2018-06-12 Institut Pasteur De Montevideo Nitroalkene trolox derivatives and methods of use thereof in the treatment and prevention of inflammation related conditions
US11400066B2 (en) 2016-10-14 2022-08-02 Institut Pasteur De Montevideo Methods of treatment of inflammation related conditions using pluripotent anti-inflammatory and metabolic modulators
US12280029B2 (en) 2016-10-14 2025-04-22 Institut Pasteur De Montevideo Methods of treatment of inflammation related conditions using pluripotent anti-inflammatory and metabolic modulators
US12006319B2 (en) 2018-05-25 2024-06-11 Cardurion Pharmaceuticals, Inc. Monohydrate and crystalline forms of 6-[(3S,4S)-4-methyl-1-(pyrimidin-2-ylmethyl)pyrrolidin-3-yl]-3-tetrahydropyran-4-yl-7H-imidazo[1,5-a]pyrazin-8-one
US12466832B2 (en) 2018-05-25 2025-11-11 Cardurion Pharmaceuticals, Inc. Monohydrate and crystalline forms of 6-[(3S,4S)-4-methyl-1-(pyrimidin-2-ylmethyl)pyrrolidin-3-yl]-3-tetrahydropyran-4-yl-7H-imidazo[1,5-a]pyrazin-8-one
US12213975B2 (en) 2018-08-31 2025-02-04 Cardurion Pharmaceuticals, Inc. PDE9 inhibitors for treating sickle cell disease
WO2020058917A1 (fr) * 2018-09-20 2020-03-26 Institut Pasteur De Montevideo Procédés de traitement d'états associés à une inflammation à l'aide de modulateurs anti-inflammatoires et métaboliques pluripotents

Similar Documents

Publication Publication Date Title
WO2015073527A1 (fr) Nitroalcène-tocophérols et analogues de ceux-ci utilisables dans le cadre du traitement et de la prévention d'affections associées à l'inflammation
AU2019222911B2 (en) Berberine salts, ursodeoxycholic salts and combinations, methods of prepration and application thereof
US20250312461A1 (en) Lymph directing prodrugs
JP7033642B2 (ja) 選択的ｓ１ｐ１レセプターアゴニストを含む薬学的合剤
KR102014524B1 (ko) 산화성 망막 질환
US20180297973A1 (en) Nitroalkene Trolox Derivatives and Methods of Use Thereof In The Treatment And Prevention of Inflammation Related Conditions
JP2012513774A (ja) ニトロ脂肪酸を含有する栄養補助物
EP3180349A1 (fr) Promédicaments de transport vers le système lymphatique
JP2015508065A (ja) 抗炎症薬としての脂肪酸
JP2013534930A (ja) 糖尿病を治療するための多成分調剤
WO2019079677A1 (fr) Procédés et compositions relatifs au 5-(1,1-diméthylheptyl)-résorcinol ultrapur
US20230321022A1 (en) Reversibly protected thiolated electrophilic fatty acids as prodrugs
US11491128B2 (en) Reversible nitroxide derivatives of nitroalkenes that mediate nitrosating and alkylating reactions
WO2011153353A1 (fr) Dérivés de tocophérol et leurs méthodes d'utilisation
JP2002532543A (ja) 抗アテローム硬化剤としてのプロアントシアニジンａ２リン脂質複合体
EP3176164A1 (fr) Isomère optique de dérivé de 1,4-benzothiazépine-1-oxyde, et composition pharmaceutique préparée à l'aide de celui-ci
CN107362159A (zh) 呫酮类化合物及其衍生物在制备降血脂药物中的应用
Shchepinov et al. Oxidative retinal diseases
HK40068487A (zh) 定向淋巴的前药
HK40035235A (en) Oxidative retinal diseases
HK1195018B (en) Oxidative retinal diseases
HK1195018A (en) Oxidative retinal diseases

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: 14861555

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: 14861555

Country of ref document: EP

Kind code of ref document: A1