[go: up one dir, main page]

WO2025016521A1 - Composé pour le traitement de l'amyloïdose à transthyrétine - Google Patents

Composé pour le traitement de l'amyloïdose à transthyrétine Download PDF

Info

Publication number
WO2025016521A1
WO2025016521A1 PCT/EP2023/025333 EP2023025333W WO2025016521A1 WO 2025016521 A1 WO2025016521 A1 WO 2025016521A1 EP 2023025333 W EP2023025333 W EP 2023025333W WO 2025016521 A1 WO2025016521 A1 WO 2025016521A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
ttr
pitb
tolcapone
amyloidosis
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.)
Pending
Application number
PCT/EP2023/025333
Other languages
English (en)
Inventor
Salvador Ventura Zamora
Irantzu Pallarès Goitiz
Francisca PINHEIRO
David REVERTER CENDRÓS
Nathalia VAREJAO
Ramon ALIBÉS ARQUÉS
Félix BUSQUÉ SÁNCHEZ
Adrià SÁNCHEZ MORALES
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.)
Universitat Autonoma de Barcelona UAB
Original Assignee
Universitat Autonoma de Barcelona UAB
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 Universitat Autonoma de Barcelona UAB filed Critical Universitat Autonoma de Barcelona UAB
Priority to PCT/EP2023/025333 priority Critical patent/WO2025016521A1/fr
Publication of WO2025016521A1 publication Critical patent/WO2025016521A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C205/00—Compounds containing nitro groups bound to a carbon skeleton
    • C07C205/45—Compounds containing nitro groups bound to a carbon skeleton the carbon skeleton being further substituted by at least one doubly—bound oxygen atom, not being part of a —CHO group
    • 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
    • A61K31/12—Ketones
    • A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system

Definitions

  • the present invention relates to the field of small molecules for their use in the treatment of transthyretin amyloidosis.
  • M-23 is a transthyretin (TTR) kinetic stabilizer that displays affinity for both binding sites and binds to TTR in human plasma with higher TTR stabilizing activity in comparison with tafamidis and tolcapone molecules, two therapeutically relevant molecules for the treatment of transthyretin amyloidosis.
  • TTR transthyretin
  • Amyloid diseases encompass a diverse range of disorders characterized by the accumulation of amyloid fibrils, highly organized structures that arise from the misfolding and subsequent aggregation of proteins. Amyloids build up in specific organs and tissues, interfering with their normal function.
  • Transthyretin amyloidosis is a group of life-threatening systemic disorders caused by the extracellular deposition of TTR amyloid fibrils. ATTR amyloidosis can be either hereditary or sporadic (non-hereditary).
  • TTR is a 55 kDa homotetrameric protein, which is mainly synthesized in the liver, choroid plexus and retinal pigment epithelium. Besides its well-known role as a transporter of thyroxine (T4) and retinol binding protein-vitamin A complex, TTR is increasingly recognized as having a neuropro- tective activity in the central nervous system.
  • T4 thyroxine
  • retinol binding protein-vitamin A complex TTR is increasingly recognized as having a neuropro- tective activity in the central nervous system.
  • TTR variants are more amyloidogenic than the wild-type protein.
  • ATTR The most common forms of ATTR are related with the aggregation of TTR mutants and inherited in an autosomal dominant manner. Most of the genetic variants causes familial amyloid polyneuropathy (FAP) and familial amyloid cardiomyopathy (FAC), e.g., V30M-TTR and V122I-TTR mutations/vari- ants. In some rare cases, TTR mutations can induce leptomeningeal amyloidosis. ATTR can also be caused by the wild-type (WT) protein, known as senile systemic amyloidosis (SSA).
  • WT wild-type
  • SSA senile systemic amyloidosis
  • Amyloid deposits start to form and then build up until they cause clinical disease, mainly affecting the nerves and/or heart, and sometimes the kidneys, eyes and synovial tissues (tendons and ligaments). Symptoms may appear at any time from early adult life onwards. This condition runs in families.
  • TTR 5 clinically relevant TTR variants V30M and V122I.
  • PITB greatly increases the stability of TTR tetramers in vitro, inhibiting their aggregation.
  • PITB presents a higher binding selectivity and stabilization potency in plasma.
  • the high-resolution crystal structures of TTR:PITB complexes have confirmed that PITB keeps critical interactions which might underlie its increased efficacy in plasma relative to tolcapone. See EXAMPLE 2 to EXAMPLE 6 (FIG.
  • cytotoxicity assays suggest a low risk associated with PITB administration (see EXAMPLE 7, FIG. 5), thereby being a drug potentially without side effects such severe liver injury. Since tolcapone is contraindicated for patients with liver-related diseases, PITB would be a perfect candidate for these patients.
  • FIG. 3 shows the crystal structures of WT-TTR (A), V30M-TTR (B) and V122I-TTR (C) in complex
  • Another aspect of the invention is a process for preparing the compound of formula (I),
  • the process for preparing the compound of formula (I) further comprises the coupling reaction of a Grignard reagent of compound (l)-2: with 3,4-dimethoxy-5-nitrobenzaldehyde, to obtain compound of formula (l)-1 :
  • the Grignard reagent of the iodoarene compound (l)-2 is prepared by an iodine-magnesium exchange reaction with i-PrMgBr at about -40 °C in an organic solvent, such as, e.g., tetrahydrofuran (THF), e.g., for about 1 hour.
  • an organic solvent such as, e.g., tetrahydrofuran (THF), e.g., for about 1 hour.
  • the synthesis leading to compound of formula (l)-1 comprises the coupling reaction of the prepared Grignard reagent of iodoarene compound (l)-2 to the 3,4-dimethoxy- 5-nitrobenzaldehyde e.g., in THF, overnight.
  • the synthesis leading to compound of formula (l)-1 further comprises an oxidation of compound (l)-2, particularly a Dess-Martin periodinane oxidation reaction using e.g., DMPI (Dess-Martin periodinane or DMP) and methylene dichloride (DCM, dichloromethane) anhydrous, overnight.
  • DMPI Dess-Martin periodinane or DMP
  • DCM methylene dichloride
  • the process comprises the following steps: a) compound (l)-1 is obtained by a coupling reaction of a Grignard reagent of compound (l)-2 and
  • compound (l)-1 is obtained by oxidation of compound (l)-2, particularly using a Dess-Martin peri- odinane oxidation; and c) compound (I) is obtained by demethylation of compound (l)-1 , particularly using boron tribromide in an organic solvent, more particularly the organic solvent is methylene dichloride.
  • the compound as defined in the first aspect is in the form of a pharmaceutical composition comprising appropriate amounts of acceptable excipients.
  • acceptable excipient or vehicle refers to compounds, materials, compositions and/or administration forms which are suitable for use in contact with the tissues of a subject (e.g., human) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • a subject e.g., human
  • Each carrier, excipient, vehicle, etc. must also be “acceptable” in the sense of being compatible with the other ingredients of the formulation.
  • compositions of this invention include, but are not limited to, fillers/diluents/bulking agents, binders, antiadherents, disintegrants, coatings, anti-caking agents, antioxidants, lubricants, sweeteners, flavors, colours, tensides, stabilizers, control-release agents, antimicrobial preservatives, acidifying agents, gelling agents, suspending agents, cosolvents, emulsifying agents and other classes of acceptable excipients.
  • the compound of the present invention is formulated into pharmaceutical compositions that can be
  • the presentation of the composition will be adapted to the type of administration used.
  • the composition can be presented in the form of solutions or any other form of clinically permissible administration and in a therapeutically effective amount.
  • the pharmaceutical composition can be thus formulated into solid, semisolid or liquid preparations, such as tablets, capsules, powders (such as those derived from lyophilization (freeze-drying) or air-drying), granules, solutions, suppositories, gels or microspheres.
  • the pharmaceutical composition is formulated for administration in liquid form or in solid form.
  • the pharmaceutical composition is in form of a capsule, a powder, a tablet, a pill, lozenges, sachets, sticks, or granules.
  • the composition is in form of a pill, a tablet or a capsule.
  • the pharmaceutical composition is in liquid form such as oral solutions,
  • Both inorganic and organic excipient materials are suitable as acceptable excipients or vehicles.
  • lactose, corn starch or derivatives thereof, talc, stearic acid or salts thereof can be used as carrier materials for tablets, coated tablets, coated tablets and hard gelatin capsules.
  • Suitable carriers for soft gelatin capsules are, e.g., vegetable oils, waxes, fats and semi-solid and liquid polyols.
  • Suitable carrier materials for the production of solutions and syrups are, e.g., water, polyols, sucrose, invert sugar and glucose.
  • Suitable carrier materials for injection solutions are, e.g., water, alcohols, polyols, glycerin and vegetable oils. Examples of suitable carrier materials for suppositories are natural or hardened oils, waxes, fats and semi-liquid or liquid polyols.
  • the pharmaceutical composition can be in the form of a sterile injectable preparation, e.g., a sterile injectable aqueous or oleaginous suspension.
  • a sterile injectable preparation e.g., a sterile injectable aqueous or oleaginous suspension.
  • This suspension can be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation can also be a sterile injectable solution or suspen ⁇
  • a non-toxic parenterally-acceptable diluent or solvent e.g., as a solution in 1 ,3-butanediol.
  • acceptable vehicles and solvents water, Ringer's solution and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil can be employed including synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful
  • oils such as olive oil or castor oil, especially in their polyoxyethylated versions.
  • oils such as olive oil or castor oil, especially in their polyoxyethylated versions.
  • These oil solutions or suspensions can also contain a long-chain alcohol diluent or dispersant.
  • the pharmaceutical composition is intravenous and comprises about 5% of nonionic surfactants (e.g., Cremophor®), 5% of mannitol, and/or 5% of dimethyl sulfoxide (DMSO).
  • nonionic surfactants e.g., Cremophor®
  • mannitol e.g., mannitol
  • DMSO dimethyl sulfoxide
  • the pharmaceutical composition is administered at about 1 mg/kg.
  • the administration volume is of about 5 ml/kg.
  • the pharmaceutical composition can be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions.
  • carriers which are commonly used include lactose and corn starch.
  • Lubricating agents such as magnesium stearate, are also typically added.
  • useful diluents include lactose and dried corn starch.
  • sweetening, flavouring or colouring agents can also be added.
  • the pharmaceutical composition is oral and comprises about 0.5% of carboxymethyl cellulose (CMC) and/or about 0.1 % of polysorbate 80 (Tween 80).
  • CBD carboxymethyl cellulose
  • Tween 80 polysorbate 80
  • the pharmaceutical composition is administered at about 10 mg/kg.
  • the pharmaceutical composition can be administered in the form of suppositories for
  • the pharmaceutical composition can be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers.
  • Carriers for topical administration of the compound of the present invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water.
  • the pharmaceutical compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers.
  • Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
  • the pharmaceutical composition can be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, particularly, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzalkonium chloride.
  • the pharmaceutical compositions can be formulated in an ointment such as petrolatum.
  • compositions can also be administered by nasal aerosol or inhalation through the use of a nebulizer, a dry powder inhaler or a metered dose inhaler.
  • a nebulizer a dry powder inhaler or a metered dose inhaler.
  • Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and can be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters
  • the compound strongly interacts with the T4 binding sites of the tetramer, with an affinity of 16, 36 and 14 nM for WT-, V30M- and V122I-TTR, respectively, which is significantly better than that of tolcapone. Furthermore, the compound shows a low risk of cytotoxicity and a favorable PK profile in vivo, with a 10.1 h half-life and 85.1 % oral bioavailability.
  • Another aspect of the invention relates to a compound as defined in the first aspect of the invention for use as a medicament.
  • an aspect of the invention refers to the use of a compound as defined in the first aspect of the present invention for the manufacture of a medicament for the treatment of a transthyretin
  • ATTR amyloid polyneuropathy
  • FAP familial amyloid polyneuropathy
  • - FAC familial amyloid cardiomyopathy
  • TTR mutations can induce leptomeningeal amyloidosis, which is often accompanied by ocular involvement.
  • V30M-TTR mutation The most common amyloidogenic variant worldwide is the V30M-TTR mutation, which is associated with FAP. It has been identified as the predominant mutation in several populations affected by transthyretin amyloidosis, including Portugal, Sweden, and some Japanese populations. V122I-TTR mutation is also relatively common, particularly in the United States, especially in individuals of African descent or West Africa, and is associated with FAC.
  • Familial amyloid polyneuropathy is disclosed e.g., in the review article of V. Pante-Bordeneuve et al., 2011 .
  • Familial amyloid cardiomyopathy is disclosed e.g., in C. Rapezzi et al., 2010.
  • Leptomeningeal amyloidosis is disclosed e.g., in H. Goren and MC. Steinberg, 1980.
  • ATTR can also be caused by the wild-type (WT) protein.
  • WT-TTR wild-type
  • the transthyretin amyloidosis is selected from the group consisting of familial amyloid polyneuropathy (FAP), senile systemic amyloidosis, familial amyloid cardiomyopathy (FAC), and leptomeningeal amyloidosis.
  • FAP familial amyloid polyneuropathy
  • FAC familial amyloid cardiomyopathy
  • leptomeningeal amyloidosis leptomeningeal amyloidosis.
  • the transthyretin amyloidosis is an hereditary amyloidosis, particularly, familial
  • FAP familial amyloid cardiomyopathy
  • FAC familial amyloid cardiomyopathy
  • the transthyretin amyloidosis is familial amyloid polyneuropathy (FAP). In another embodiment, the transthyretin amyloidosis is senile systemic amyloidosis. In another em ⁇
  • the transthyretin amyloidosis is familial amyloid cardiomyopathy (FAC).
  • FAC familial amyloid cardiomyopathy
  • the transthyretin amyloidosis is leptomeningeal amyloidosis.
  • the ATTR is associated with a mutation in transthyretin (TTR) protein/gene (i.e. , amyloidogenic TTR variants).
  • TRR transthyretin
  • the TRR mutation is selected from mutations listed
  • the familial amyloid polyneuropathy is associated with V30M, V28M, L55P, L58H and/or S77Y transthyretin mutations/variants. In a particular embodiment, the familial amyloid polyneuropathy is associated with V30M transthyretin variant.
  • the familial amyloid cardiomyopathy is associate with V122I, T60A, I68L and/or L1 11 M transthyretin mutations/variants. In a particular embodiment, the familial amyloid cardiomyopathy is associated with V122I transthyretin variant.
  • S77Y mutation is one of the most frequent mutations in France (apart from V30M); L55P is the most aggressive TTR variant; I68L is the most frequent mutation in Italy; and L111 M is the most frequent in Denmark.
  • treat refers to a clinical intervention to prevent
  • a disease or condition e.g., suppress or inhibit
  • cure the disease or condition e.g., delay the onset of the disease or condition; reduce the seriousness or severity of the disease or condition; ameliorate or eliminate one or more symptoms or sequelae associated with a disease or condition; or the provision of beneficial effects to a subject with a disease or condition, without necessarily curing the disease or condition.
  • the term refers to a clinical intervention to improve one or more symptoms; improve one or more sequelae; prevent (e.g., suppress, inhibit or delay) one or more symptoms; prevent (e.g., suppress, inhibit or delay) one or more sequelae; delay one or more symptoms; delay one or more sequelae; ameliorate one or more symptoms; ameliorate one or more sequelae; shorten the duration of one or more symptoms; shorten the duration of one or more sequelae; reduce the frequency of one or more symptoms; reduce the frequency of one or more sequelae; reduce the severity of one or more symptoms; reduce the severity of one or more sequelae; improve the quality of life; increase survival; prevent (e.g., suppress, inhibit or delay) a recurrence of the disease or condition; delay a recurrence of the disease or condition; reduce the severity of the disease; or any
  • treatment also includes prophylaxis or prevention (e.g., suppression, inhibition or delay) of a disease or condition or its symptoms or sequelae thereof.
  • prophylaxis refers to a therapeutic or
  • 10 refer to any mammalian subject, including without limitation, humans, domestic animals (e.g., dogs, cats and the like), farm animals (e.g., cows, sheep, pigs, horses and the like), and laboratory animals (e.g., monkey, rats, mice, rabbits, guinea pigs and the like) for whom diagnosis, treatment, or therapy is desired, particularly humans.
  • domestic animals e.g., dogs, cats and the like
  • farm animals e.g., cows, sheep, pigs, horses and the like
  • laboratory animals e.g., monkey, rats, mice, rabbits, guinea pigs and the like
  • the subject is a mammal, including a human. In a further particular embodiment, the subject is a human subject.
  • the subject presents a mutation in the transthyretin (TTR) protein/gene.
  • TRR transthyretin
  • the TRR mutation is selected from mutations listed in http://amyloidosismutations.com/mut- attr.php.
  • the subject presents:
  • a mutant TTR comprising at least one amino acid substitution at a position in the sequence of TTR (Uniprot P02766, version 1 , date June 28, 2023) selected from the group consisting of positions
  • ATTR The symptoms and sequelae of ATTR can vary depending on the type and progression of the disease. Some common symptoms and sequelae associated with ATTR are:
  • peripheral neuropathy e.g., numbness, tingling, and weakness in the extremities, starting in the lower limbs and progressing upwards. Loss of sensation and motor function may lead to difficulty in walking or performing fine motor tasks;
  • - autonomic dysfunction impaired regulation of involuntary bodily functions, such as blood pressure, heart rate, digestion, and sweating. Symptoms can include orthostatic hypotension (low blood pressure upon standing), gastrointestinal disturbances, sexual dysfunction, and abnormal sweating;
  • cardiomyopathy which is characterized by the stiffening and thickening of the
  • Symptoms can include shortness of breath, fatigue, chest pain, palpitations, and eventually heart failure;
  • gastrointestinal tract in some cases of ATTR, the gastrointestinal tract can be affected, leading to symptoms such as abdominal pain, diarrhea, constipation, and weight loss;
  • deposition of amyloid fibrils in the eye can cause various ocular abnormalities, including dry eyes, blurred vision, and vitreous opacities;
  • ATTRwt in particular, can result in the deposition of amyloid fibrils in the kidneys, leading to renal impairment and potentially progressing to end-stage renal disease;
  • ATTR can involve the central nervous sys ⁇
  • amyloid deposition can affect other organs, such as the liver, lungs, and spleen, although the severity and impact vary.
  • the main symptoms and sequelae of ATTR for each type of ATTR can be:
  • FAP familial amyloid polyneuropathy
  • FAC familial amyloid cardiomyopathy
  • the administration of the compound as defined in the first aspect of the invention results in at least one outcome (i.e., effect) selected, but not limited to, from the group consisting of:
  • TTR transthyretin
  • cardiac function e.g., slowing or halting the progression of cardiomyopathy, or improved heart muscle function
  • - reduced risk of heart failure e.g., prevention or delay of the onset of heart failure
  • renal impairment e.g., delay or prevention end-stage renal disease
  • the invention also refers to a compound as defined in the first aspect of the invention in combination with one or more therapeutic agents, particularly for the treatment of a transthyretin amyloidosis.
  • the additional therapeutic agent is selected from the group consisting of
  • 10 can be administered in a single pharmaceutical composition, or in two different pharmaceutical compositions, either administered simultaneously, sequentially or separately after a certain period of time. Particularly, if the administration is not simultaneous, the compounds are administered in a relatively close time proximity to each other. Furthermore, compounds are administered in the same or different dosage form or by the same or different administration route, e.g., one compound can be administered intravenously and the other compound can be administered orally.
  • the term "combination” relates herein to the various combinations of the compound as defined in the first aspect and other therapeutic agent(s), e.g., in a single pharmaceutical composition, in a combined mixture composed from separate pharmaceutical formulations/compositions of the single active compounds, such as a "tank-mix", and in a combined use of the single active ingredients when applied in a sequential manner, i.e. one after the other with a reasonably short period, such as a few hours or days or in simultaneous administration.
  • the order of applying the compounds is not essential.
  • the compound as defined in the first aspect is independently administered from the other therapeutic agent(s) (i.e., in two units) but at the same time.
  • the compound of the present invention is administered first and then the other therapeutic agent(s) is separately or sequentially administered; alternatively, the other therapeutic agent(s) is administered first and then the compound of the present is separately or sequentially administered.
  • Tolcapone was purchased from Fisher.
  • PITB (I) was chemically synthesized as shown in Scheme I. The detailed synthesis procedure is described below.
  • the cDNA encoding for WT-TTR was cloned into a pET28a vector (Novagen).
  • the vectors encoding for V30M- and V122I-TTR were prepared by classical site-directed mutagenesis protocols using the WT-TTR vector as a template.
  • WT-, V30M- and V122I-TTR were expressed in Escherichia coli BL21 (DE3) and purified as previously described (F. Pinheiro et al., 2021). The purest fractions eluting from the gel filtration chromatography were combined and kept at -20 °C until use.
  • the concentration of protein was determined spectrophotometrically at 280 nm, using a molar extinction coefficient of 77 600 M- 1 cm 1 .
  • the Trp residues Upon denaturation, the Trp residues become more exposed to the polar solvent, which is accompanied by a change in the emission maximum from ⁇ 335 to ⁇ 355 nm.
  • the fluorescence intensity ratio 355/335 nm was calculated for each time point and the values normalized from minimum (folded state) to maximum (unfolded state), with the maximum being the one of the control sample after 96 h incubation.
  • TTR solutions (7.2 pM in 10 mM sodium phosphate, 100 mM KCI, 1 mM EDTA, 1 mM DTT, pH 7.0) were incubated with varying concentrations of test compound for 30 min at 37 °C. The percentage of DMSO was the same in all samples (5%).
  • each inhibitor concentration was calculated by dividing the turbidity of the test sample by that of a sample aggregated in the absence of compound, and multiply by 100.
  • Cocrystals of WT-TTR/PITB, V30M-TTR/PITB, V122I-TTR/PITB and V30M-TTR/tolcapone were obtained as explained previously (F. Pinheiro et al., 2021). Briefly, purified proteins (140 pM) were mixed with 1 .4 mM of PITB/tolcapone and cocrystallized at 18 °C by hanging-drop vapor diffusion methods. The reservoir solution contained 20-30% PEG 400, 200 mM CaCh, 100 mM HEPES, pH 7.0-8.0.
  • the crystals were flash-frozen in liquid nitrogen (100 K) and diffraction data were collected at the BL13-XALOC beamline from the ALBA Synchrotron in Barcelona. Data were integrated and merged using XDS and scaled, reduced, and further analyzed using CCP4.
  • the structures of TTR/PITB and V30M-TTR/tolcapone complexes were determined from the X-ray data by molecular replacement with Phenix (version 1.19.2-4158) using a previous TTR structure (PDB 1 F41) as a model. Model refinement and building were done with Phenix and Coot, respectively.
  • T4-TBG globulin
  • T4-ALB albumin + T4-TTR
  • TTR tetramer + monomer The ratio of the TTR tetramer over total TTR (TTR tetramer + monomer) was determined for each sample, and the percentage of tetramer stabilization was calculated as ((ratio treated sample - ratio control sample)/ratio control sample) x 100.
  • HeLa and HepG2 cells were cultured in MEM ALPHA medium (Gibco) supplemented with 10% fetal bovine serum at 37 °C in a 5% CO2 humidified atmosphere. HeLa cells were seeded at 3500 cells/well and HepG2 cells at 4500 cells/well in 96-well plates and incubated with increasing concentrations of compound (2-100 pM) for 72 h at 37°C. Controls were prepared with the equivalent amount of DMSO relative to each concentration of compound. Then, 10 pl of PrestoBlue® reagent (Thermo Fisher Scientific) were added to each well, and after incubating for 15 min at 37°C, the fluorescence emis ⁇
  • PITB intravenous, IV and oral, PO
  • tolcapone oral, PO
  • PITB was dissolved in 5% Cremophor/5% Mannitol/5% DMSO.
  • the compound was administered at 1 mg/Kg, with an administration volume of 5 mL/Kg.
  • timepoints after administration 0.0833, 0.5, 1 , 2, 4, 6 and 24 h
  • three animals were anesthetized with isoflurane,
  • t 0 h (predose). Blood samples were centrifuged at 10,000 rpm for 5 min to obtain the plasmas, which were stored at -80 °C until analysis.
  • PITB and tolcapone were formulated in 0.5% CMC/0.1 % tween 80 and admin ⁇
  • Plasma samples were obtained as described above at 0.25, 0.5, 1 , 2, 4, 6 and 24 h post-dosing (three animals for time point). Three animals were used as blank with no administration (predose). All plasma samples were analyzed using the API 3200 LC-MS/MS system (Sciex) coupled to an UPLC-Acquity (Waters). A calibration curve was done for each compound in plasma. The lower limit of quantification (LLOQ) for PITB and
  • 10 tolcapone were 7.68 ng/mL and 2.73 ng/mL, respectively.
  • PK parameters were determined with a non-compartmental model using Phoenix 64 8.3 (WinNolin) software.
  • TTR subunits do not unfold until the tetramer dissociates. Since monomer unfolding is much faster than tetramer dissociation, it is possible to evaluate the rate of tetramer dissociation by irreversibly linking these two processes. This can be achieved by using urea concentrations in the post-transition zone for tertiary structural changes, which assures that monomers unfold extremely fast and cannot
  • WT-TTR and the two most clinically important TTR variants, V30M- and V122I- TTR (1.8 pM) were incubated in the absence or presence of PITB (3.6 pM) and TTR denaturation was initiated by adding urea to a final concentration of 6 M.
  • Tryptophan (Trp) intrinsic fluorescence was measured each 24 h and used to calculate the fraction of unfolded protein (FIG. 1 (A)). The effect of PITB was compared to the one of tolcapone.
  • PITB stabilizing activity was equivalent to the one exerted by tolcapone for each of the analyzed proteins.
  • EXAMPLE 3 PITB protects TTR from aggregation in vitro
  • TTR samples were mixed with
  • PITB is highly effective at inhibiting the aggregation of WT-TTR and, most importantly, of V30M- and V122I-TTR.
  • TTR aggregation >60% at a 1 :1 TTR/PITB ratio and >87.5% when PITB concentration is higher than or equal to the one of T4-binding sites.
  • PITB performed better than tolcapone, inhibiting up to 89.2% and 92% the aggregation of WT-TTR and V30M-TTR, respectively.
  • V122I-TTR PITB equaled the potency of tolcapone, reaching up to 90% inhibition at 20 pM.
  • ITC isothermal titration calorimetry
  • PITB binds with high affinity and no cooperativity to both WT and TTR variants V30M and V1 121 (FIG. 2 and Table 1). Moreover, PITB binds to WT- and V122I-TTR with an affinity > 3-fold than that of tolcapone, and to V30M-TTR with an affinity > 12-fold.
  • V122I-TTR: Kd1 8.1 nM and Kd2 ⁇ 1 pM
  • the binding of PITB was entirely enthalpically driven (AH ⁇ 0; -TAS > 0), suggesting the establishment of noncovalent interactions (e.g., hydrogen bonds or ionic interactions) at the binding interface.
  • TTR:PITB complexes were determined at 1 .85 (WT-TTR:PITB), 1.20 (V30M-TTR:PITB) and 1.42 (V122I-TTR:PITB) A resolution (FIG. 3). Additionally, inventors obtained, for the first time, the cocrystal structure of tolcapone with V30M-TTR, at a resolution of 1 .57 A.
  • PITB binds to TTR T ⁇ binding sites at the weaker AB/CD dimer-dimer interface. Because of the twofold symmetry axis that runs through the binding pockets, two symmetry-related binding conformations appear for PITB. In all structures, PITB was found in the forward binding mode, with the 3- methoxy-4-hydroxy-5-nitrophenyl pointing to the outer binding cavity, where it makes hydrophobic interactions with the residues from halogen binding pockets (HBPs) 2/2’ and 1/1 ’ (FIG. 3). In this orientation, the 4-hydroxy group of PITB forms hydrogen bonds with the K15/K15’ residues of TTR, which in turn stabilize their electrostatic interactions with E54/E54’. These interactions by K15 are also observed in TTR:tolcapone complexes and were proposed to close the binding cavity, protecting the protein-ligand complex from the solvent.
  • HBPs halogen binding pockets
  • PITB establishes important interactions in the inner binding cavity, which are not present in the TTR:tolcapone structures.
  • the fluorine atom of the 3-fluoro-5-hydroxy- phenyl ring interacts with A108, while the hydroxy group forms hydrogen bonds with the S117 residues from adjacent subunits.
  • the hydrogen bonds between PITB and S117 help to bridge the dimers at the weaker dimer-dimer interface, increasing the kinetic barrier for tetramer dissociation.
  • a short hydrogen bond can be formed between the S117 residues from adjacent monomers in the same dimer (FIG. 3(D)). This inter-subunit interaction was also observed in the structure of the kinetically stabilized TTR variant T119M and could further contribute to the high stabilizing effect of PITB.
  • EXAMPLE 6 PITB selectively binds and stabilizes WT-TTR and V30M-TTR in human plasma
  • PITB must selectively bind to TTR in plasma over all other plasmatic proteins.
  • WT-TTR control individuals
  • V30M-TTR V30M-TTR
  • T4-binding proteins were detected by autoradiography, as shown in FIG. 4(C).
  • T4-binding globulin TSG
  • ALB albumin
  • TTR T4-binding globulin
  • PITB selectively binds TTR in human plasma
  • inventors studied its effect on the stability of TTR in the plasma of normal individuals and FAP V30M patients.
  • the ability of PITB to prevent tetramer dissociation was monitored ex vivo by isoelectric focusing (IEF) under partially dissociating conditions (4 M urea). Tolcapone was also used as a control. Briefly, plasma samples were incubated with or without compound overnight at 4 °C and, after IEF, the bands were compared. Under the conditions used, it is possible to distinguish two main sets of bands, one corresponding to monomers (normal and oxidized forms), and one including several bands of lower pl that represent different forms of tetramers (FIG. 4(D)).
  • PITB has a higher stabilizing activity than tolcapone, especially in plasma from FAP patients, with an increase in tetramer stability
  • EXAMPLE 7 PITB displays no toxicity for human cells
  • HeLa human cervical carcinoma
  • HepG2 human hepatoblastoma
  • PITB was substantially less toxic than tolcapone at concentrations higher than 10 pM, suggesting that PITB could be a safer molecule when administered at high doses than tolcapone.
  • EXAMPLE 8 Pharmacokinetic (PK) studies in mice confirm the bioavailability and stability of PITB
  • PK profiles for PITB following IV (1 mg/Kg) or PO (10 mg/Kg) administration of the compound are shown in FIG. 6(A), and the calculated parameters are depicted in Table 2.
  • a Cmax value of 4043.3 ng/mL and a ti/2 of 3.5 h were determined. Additionally, the results demonstrated that PITB is rapidly absorbed after PO administration, reaching a Cmax of 4176.7 ng/mL, at 0.5 h (Tmax). While the concentration achieved by the oral route was very similar to that observed for the IV infusion, the elimination of PITB was considerably slower in this case, with a ti/2 of 10.1 h.
  • the concentration of PITB in plasma was still quantifiable 24 h post-dosing.
  • the area under the plasma concentration-time curve (AUC) values were 3162.3 IT ng/mL and 26895.9 h ng/ml for the IV (1 mg/Kg) and PO (10 mg/Kg) administration of PITB, respectively.
  • AUC area under the plasma concentration-time curve
  • %F oral bioavailability
  • the bioavailability of PITB is significantly higher than the one reported for AGI O (26.84%), a TTR kinetic stabilizer that is under clinical trials for ATTR- related cardiomyopathy.
  • PITB seems to have better PK proper ⁇ ties than tolcapone, which might be translated into an improved pharmacologic activity in vivo.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Epidemiology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne un composé de benzophénone de formule (I) et ses sels pharmaceutiquement acceptables, qui sont utiles dans le traitement d'une amyloïdose à transthyrétine. Le composé présente une affinité plus élevée vis-à-vis de la transthyrétine, une biodisponibilité plus élevée et une toxicité réduite. L'invention concerne également un procédé de préparation du composé et une composition pharmaceutique comprenant le composé.
PCT/EP2023/025333 2023-07-18 2023-07-18 Composé pour le traitement de l'amyloïdose à transthyrétine Pending WO2025016521A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/EP2023/025333 WO2025016521A1 (fr) 2023-07-18 2023-07-18 Composé pour le traitement de l'amyloïdose à transthyrétine

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/EP2023/025333 WO2025016521A1 (fr) 2023-07-18 2023-07-18 Composé pour le traitement de l'amyloïdose à transthyrétine

Publications (1)

Publication Number Publication Date
WO2025016521A1 true WO2025016521A1 (fr) 2025-01-23

Family

ID=87519949

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2023/025333 Pending WO2025016521A1 (fr) 2023-07-18 2023-07-18 Composé pour le traitement de l'amyloïdose à transthyrétine

Country Status (1)

Country Link
WO (1) WO2025016521A1 (fr)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0237929A1 (fr) 1986-03-11 1987-09-23 F. Hoffmann-La Roche Ag Dérivés du pyrocatéchol substitués en 3,5

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0237929A1 (fr) 1986-03-11 1987-09-23 F. Hoffmann-La Roche Ag Dérivés du pyrocatéchol substitués en 3,5

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
"Uniprot", Database accession no. P02766
C. RAPEZZI ET AL.: "Transthyretin-related amyloidosis and the heart: a clinical overview", NAT. REV, CARDIOL., vol. 7, 2010, pages 398 - 408
F. PINHEIRO ET AL.: "Development of a Highly Potent Transthyretin Amyloidogenesis Inhibitor: Design, Synthesis, and Evaluation", J. MED. CHEM., vol. 65, no. 21, 28 October 2022 (2022-10-28), pages 14673 - 14691
F. PINHEIRO ET AL.: "Tolcapone, a potent aggregation inhibitor for the treatment of familial leptomeningeal amyloidosis", THE FEBS JOURNAL, vol. 288, 2021, pages 310 - 324
H. GORENMC. STEINBERG: "Familial oculoleptomeningeal amyloidosis", BRAIN, vol. 103, 1980, pages 473 - 495
P. WESTERMARK ET AL.: "Fibril in senile systemic amyloidosis is derived from normal transthyretin", PROC. NATL. ACAD. SCI. USA, vol. 87, 1990, pages 2843 - 5, Retrieved from the Internet <URL:http://amyloidosismutations.com/mut-attr.php>
PINHEIRO FRANCISCA ET AL: "Development of a Highly Potent Transthyretin Amyloidogenesis Inhibitor: Design, Synthesis, and Evaluation", JOURNAL OF MEDICINAL CHEMISTRY, vol. 65, no. 21, 10 November 2022 (2022-11-10), US, pages 14673 - 14691, XP093126570, ISSN: 0022-2623, DOI: 10.1021/acs.jmedchem.2c01195 *
V. PANTE-BORDENEUVE ET AL.: "Familial amyloid polyneuropathy", LANCET NEUROL., vol. 10, no. 12, 2011, pages 1086 - 97

Similar Documents

Publication Publication Date Title
JP6938705B2 (ja) 多発性嚢胞腎の予防及び／又は治療薬
CN110305095A (zh) 色甘酸衍生物以及成像和治疗的相关方法
EA028378B1 (ru) Дейтерированные производные ивакафтора
TW202425973A (zh) 使用ag10治療ttr澱粉樣沉積症之方法
JP2024531374A (ja) タキソールコンジュゲート化合物、これを含む医薬組成物およびそれらを使用する方法
CN103781771B (zh) 苯并噻唑酮化合物
WO2025016521A1 (fr) Composé pour le traitement de l&#39;amyloïdose à transthyrétine
EP3609495B1 (fr) Association comprenant un antagoniste de nk1 et méthode de traitement de synucléinopathies
JP2008179541A (ja) 神経因性疼痛治療薬
JP7569953B2 (ja) Ｓｓｔｒ４アゴニスト及びその塩の調製のための方法並びに使用のための用量レジメン
EP2961391B1 (fr) Préparation contenant un composé de benzothiazolone
CN113260612B (zh) 用于治疗或预防肌营养不良症的化合物及其在医药用途中的治疗、改善或预防用衍生物
JP2013010776A (ja) 脊髄小脳変性症治療剤
CN112457291B (zh) 苯并硫代吡喃酮类化合物的盐及其制备方法和用途
JP7228529B2 (ja) １，４－ベンゾチアゼピン－１－オキシド誘導体の光学異性体を含む腎機能障害の改善薬
JP4243701B2 (ja) ベンズアミド誘導体を有効成分とするリウマチ治療薬
EP2890687B1 (fr) Sels de composé benzothiazolone utilisés en tant qu&#39;agoniste de beta-2-adrenoceptor
US20250197371A1 (en) N-acylhydrazonic compounds, use in the treatment of amyloid and non-amyloid degenerative aggregopathies, and pharmaceutical composition
CN102215842A (zh) 含有吡啶-3-甲醛o-(哌啶-1-基-丙基)-肟衍生物作为有效成分的脉络膜视网膜变性疾病的治疗剂
JPH06192089A (ja) メイラード反応阻害剤
US20230134773A1 (en) N-acylhydrazonic compounds, use in the treatment of amyloid and non-amyloid degenerative aggregopathies, and pharmaceutical composition
JP2025526688A (ja) 疾患を治療するためのモノカルボン酸トランスポータ４の複素環式阻害剤の塩
JPH01501797A (ja) ３―置換―１―アルキルアミノ―２―プロパノールの不安定なケトン誘導体と、そのβ―アドレナリン作動効果遮断薬としての応用
HK1212213B (en) Formulation comprising benzothiazolone compound
HK1242680A1 (en) Salts of benzothiazolone compound as beta-2-adrenoceptor agonist

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: 23748426

Country of ref document: EP

Kind code of ref document: A1