WO2023240205A1 - Composés deutérés - Google Patents

Composés deutérés Download PDF

Info

Publication number
WO2023240205A1
WO2023240205A1 PCT/US2023/068152 US2023068152W WO2023240205A1 WO 2023240205 A1 WO2023240205 A1 WO 2023240205A1 US 2023068152 W US2023068152 W US 2023068152W WO 2023240205 A1 WO2023240205 A1 WO 2023240205A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
deuterium
pharmaceutically acceptable
acceptable salt
incubation
Prior art date
Application number
PCT/US2023/068152
Other languages
English (en)
Inventor
Evan Smith
Romain Siegrist
Original Assignee
Neurocrine Biosciences, Inc.
Idorsia Pharmaceuticals Ltd
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 Neurocrine Biosciences, Inc., Idorsia Pharmaceuticals Ltd filed Critical Neurocrine Biosciences, Inc.
Publication of WO2023240205A1 publication Critical patent/WO2023240205A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
    • C07B59/002—Heterocyclic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/12—Drugs for disorders of the metabolism for electrolyte homeostasis
    • A61P3/14—Drugs for disorders of the metabolism for electrolyte homeostasis for calcium homeostasis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/05—Isotopically modified compounds, e.g. labelled

Definitions

  • the present disclosure relates to deuterated compounds and their use as T-type calcium channel blockers in the treatment or prevention of various diseases or disorders associated with calcium T channels.
  • Intracellular calcium concentrations control important life processes such as signal transduction pathways, hormones and neurotransmitter release, muscular contraction, gene expression and cell division.
  • Control of calcium influx across the cellular membrane is in part regulated by a family of transmembrane proteins tenned voltage-gated calcium channels (VOCs). They are activated by changes in electrical potential difference across the membrane and have been further classified into different subtypes based on biophysical and pharmacological considerations: Cavl.x (L-type for Long-lasting), Cav2.x (N-, P/Q- and R-types; N for Neuronal, P for Purkinje cells, Q (after P) and R for Remaining or Resistant) and Cav3.x (T-type for Transient).
  • VOCs voltage-gated calcium channels
  • the L, N, P and Q-type channels activate at more positive potentials (high voltage activated) and display diverse kinetics and voltage-dependent properties.
  • the T-type class (or “low voltage-activated”) is characterized by fast inactivation (transient) and small conductance (tiny) and is composed of three members due to the different main pore-forming al subunits: Cav3.1 (al G), Cav3.2 (al H) and Cav3.3 (al I).
  • the compounds of the present disclosure are calcium T channel blockers and therefore useful for the prevention or treatment of diseases or disorders where calcium T channels are involved.
  • the present application provides, inter alia, a compound of Formula I: or a pharmaceutically acceptable salt thereof, wherein the constituent members are defined herein.
  • compositions comprising a compound described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
  • the present disclosure further provides methods of blocking a T-type calcium channel, comprising contacting the T-type calcium channel with a compound described herein, or a pharmaceutically acceptable salt thereof.
  • the present disclosure further provides methods of blocking a T-type calcium channel in a patient, comprising administering to the patient a compound described herein, or a pharmaceutically acceptable salt thereof.
  • the present disclosure further provides methods of treating a disease or disorder associated with a T-type calcium channel in a patient, comprising administering to the patient a compound described herein, or a pharmaceutically acceptable salt thereof.
  • the present disclosure further provides compounds described herein, or a pharmaceutically acceptable salt thereof, for use in any of the methods described herein.
  • the present disclosure further provides uses of a compound described herein, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for use in any of the methods described herein. DESCRIPTION OF DRAWINGS
  • FIG. 1 shows recovery of total radioactivity in incubations of Compound 2 with liver microsomes and hepatocytes of various species.
  • FIG. 2 shows proposed metabolic pathways of Compound 1.
  • the aglycon M29 was only detected after enzymatic cleavage of the corresponding glucuronic acid conjugate M4.
  • FIG. 3 shows cross-species comparison of Compound 2 metabolic profdes after 60 minutes incubation with liver microsomes. All values are expressed as percent of total chromatogram radioactivity and rounded to two significant figures. Empty cells indicate the absence of a metabolite.
  • FIG. 4 shows radiochromatogram following 60 minutes incubation of Compound 2 with human liver microsomes.
  • FIG. 5 shows radiochromatogram following 60 minutes incubation of Compound 2 with Wistar rat liver microsomes.
  • FIG. 6 shows radiochromatogram following 60 minutes incubation of Compound 2 with Beagle dog liver microsomes.
  • FIG. 7 shows radiochromatogram following 60 minutes incubation of Compound 2 with cynomolgus monkey liver microsomes.
  • FIG. 8 shows radiochromatogram following 60 minutes incubation of Compound 2 with CD-I mouse liver microsomes.
  • FIG. 9 shows radiochromatogram following 60 minutes incubation of Compound 2 with NZW rabbit liver microsomes.
  • FIGs. 10A-10C show radiochromatogram following 60 minutes incubation of Compound 2 with liver microsomes of human (FIG. 10A), rat (FIG. 10B), and mouse (FIG. 10C) in the absence of NADPH.
  • FIG. 11 shows radiochromatogram following 60 minutes incubation of Compound 2 in the absence of liver microsomes.
  • FIG. 12 shows cross-species comparison of Compound 2 metabolic profiles in incubations with hepatocytes. All values are expressed in percent of total chromatogram radioactivity and rounded to two significant figures. Empty cells indicate the absence of a metabolite.
  • FIGs. 13A-13B show radiochromatogram following 4 h (FIG. 13A) and 24 h (FIG. 13B) incubation of Compound 2 with ready-plated human hepatocytes (batch 1).
  • FIGs. 14A-14B show radiochromatogram following 4 h (FIG. 14A) and 24 h (FIG. 14B) incubation of Compound 2 with ready-plated human hepatocytes (batch 2).
  • FIGs. 15A-15B show radiochromatogram following 4 h (FIG. 15 A) and 24 h (FIG. 15B) incubation of Compound 2 with cryopreserved human hepatocytes (batch 3).
  • FIGs. 16A-16B show radiochromatogram following incubation of Compound 2 with fresh human hepatocytes (batch 4) in the absence (FIG. 16A) or presence (FIG. 16B) of P-glucuronidase.
  • FIG. 17 shows radiochromatogram following 24 h incubation of Compound 2 with Wistar rat hepatocytes.
  • FIG. 18 shows radiochromatogram following 24 h incubation of Compound 2 with Beagle dog hepatocytes.
  • FIG. 19 shows radiochromatogram following 24 h incubation of Compound 2 with cynomolgus monkey hepatocytes.
  • FIG. 20 shows radiochromatogram following 4 h incubation of Compound 2 with CD-I mouse hepatocytes.
  • FIG. 21 shows radiochromatogram following 6 h incubation of Compound 2 with NZW rabbit hepatocytes.
  • FIG. 22 shows radiochromatogram following 24 h incubation of Compound 2 in the absence of hepatocytes.
  • FIG. 23 shows stability of Compound 2 in plasma. All values are expressed in percent of total chromatogram radioactivity and rounded to two significant figures; n.d.: not determined. Empty cells: 0.
  • FIG. 24 shows stability of Compound 2 in blood at 37 °C. All values are expressed in percent of total chromatogram radioactivity; n.d.: not determined. Empty cells: 0. FTG. 25A-25D show representative radiochromatograms following 6 h incubation of Compound 2 with rat plasma at 37 °C (FIG. 25 A), 4 h incubation with rat plasma and 0.1 % DCV at 37 °C (FIG. 25B), 4 h incubation with rat plasma and 0.2 mM BNPP at 37 °C (FIG. 25C), and 4 h incubation with rat plasma at 4 °C (FIG. 25D).
  • FIGs. 26A-26B show representative radiochromatograms following incubation of Compound 2 with blood of human (FIG. 26A) and rat (FIG. 26B) at 37°C.
  • Compound 1 i.e., N-(l-((5-cyanopyridin-2-yl)methyl)-lH-pyrazol-3-yl)-2-(4-(l- (trifluoromethyl)cyclopropyl)phenyl)acetamide
  • T-type calcium channel blocker in development for the prevention or treatment of epilepsy (see e.g., U.S.
  • the present application provides deuterated analogs of Compound 1, and pharmaceutically acceptable salts thereof. Substitution with heavier isotopes, such as deuterium, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances, (see e.g., A. Kerekes et. al. J. Med. Chem. 2011, 54, 201-210; R. Xu et. al. J. Label Compd. Radiopharm. 2015, 58, 308-312). In particular, substitution at one or more metabolism sites may afford one or more of the therapeutic advantages. In some embodiments, the present application provides a compound of Formula I: or a pharmaceutically acceptable salt thereof, wherein:
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 15 , R 17 , and R 18 are each independently selected from hydrogen and deuterium; and wherein at least one R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 is deuterium.
  • one to eighteen of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 are deuterium, for example, one to sixteen, one to fourteen, one to twelve, one to ten, one to eight, one to six, one to four, or one to two of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 are deuterium .
  • one to six of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 are deuterium.
  • two to six of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 1(J , R u , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 are deuterium.
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 are deuterium.
  • four to six of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 are deuterium.
  • one of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 is deuterium.
  • two of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 are each deuterium.
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 are each deuterium.
  • four of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 13 , R 16 , R 17 , and R 18 are each deuterium.
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 are each deuterium.
  • R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 13 , R 16 , R 17 , and R 18 are each deuterium.
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 are each deuterium.
  • the compound of Formula I is a compound of Formula II: or a pharmaceutically acceptable salt thereof.
  • the compound of Formula I is a compound of Formula III: or a pharmaceutically acceptable salt thereof.
  • the compound of Formula I is a compound of Formula IV:
  • the compound of Formula I is a compound of Formula V: or a pharmaceutically acceptable salt thereof.
  • the compound of Formula I is selected from:
  • the compounds disclosed and described herein allow atoms at each position of the compound independently to have: 1) an isotopic distribution for a chemical element in proportional amounts to those usually found in nature or 2) an isotopic distribution in proportional amounts different to those usually found in nature unless the context clearly dictates otherwise.
  • a particular chemical element has an atomic number defined by the number of protons within the atom’s nucleus. Each atomic number identifies a specific element, but not the isotope; an atom of a given element may have a wide range in its number of neutrons. The number of both protons and neutrons in the nucleus is the atom's mass number, and each isotope of a given element has a different mass number.
  • a compound wherein one or more atoms have an isotopic distribution for a chemical element in proportional amounts different to those usually found in nature is commonly referred to as being an isotopically-labeled compound.
  • Each chemical element as represented in a compound structure may include any isotopic distribution of said element.
  • a hydrogen atom may be explicitly disclosed or understood to be present in the compound.
  • the hydrogen atom can be an isotopic distribution of hydrogen, including but not limited to protium ( 1 H) and deuterium ( 2 H) in proportional amounts to those usually found in nature and in proportional amounts different to those usually found in nature.
  • references herein to a compound encompasses all potential isotopic distributions for each atom unless the context clearly dictates otherwise.
  • isotopes include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, chlorine, bromine, and iodine.
  • any of the compounds as disclosed and described herein may include radioactive isotopes.
  • isotopes of hydrogen include protium ( 1 H), deuterium ( 2 H), and tritium ( 3 H).
  • Isotopes of carbon include carbon-11 ( U C), carbon-12 ( 12 C), carbon-13 ( 13 C), and carbon-14 ( 14 C).
  • Isotopes of nitrogen include nitrogen-13 ( 13 N), nitrogen-14 ( 14 N) and nitrogen- 15 ( 15 N).
  • Isotopes of oxygen include oxygen- 14 ( 14 O), oxygen- 15 ( 15 O), oxygen- 16 ( 16 O), oxygen- 17 ( 17 O), and oxygen- 18 ( 18 O).
  • Isotope of fluorine include fluorine-17 ( 1 Z F), fluorine-18 ( 18 F) and fluorine-19 ( 19 F).
  • Isotopes of phosphorous include phosphorus-31 ( 31 P), phosphorus-32 ( 32 P), phosphorus-33 ( 33 P), phosphorus-34 ( 34 P), phosphorus-35 ( 35 P) and phosphorus-36 ( 36 P).
  • Isotopes of sulfur include sulfur-32 ( 32 S), sulfur-33 ( 33 S), sulfur-34 ( 34 S), sulfur-35 ( 35 S), sulfur-36 ( 36 S) and sulfur-38 ( 38 S).
  • Isotopes of chlorine include chlorine-35 ( 35 C1), chlorine-36 ( 36 C1) and chlorine-37 ( 37 C1).
  • Isotopes of bromine include bromine-75 ( 75 Br), bromine-76 ( 76 Br), bromine-77 ( 77 Br), bromine-79 ( 79 Br), bromine-81 ( 81 Br) and bromine-82 ( 82 Br).
  • Isotopes of iodine include iodine-123 ( 123 I), iodine-124 ( 124 I), iodine-125 ( 125 I), iodine-131 ( 131 I) and iodine-135 ( 135 I).
  • atoms at every position of the compound have an isotopic distribution for each chemical element in proportional amounts to those usually found in nature.
  • an atom in one position of the compound has an isotopic distribution for a chemical element in proportional amounts different to those usually found in nature (remainder atoms having an isotopic distribution for a chemical element in proportional amounts to those usually found in nature).
  • atoms in at least two positions of the compound independently have an isotopic distribution for a chemical element in proportional amounts different to those usually found in nature (remainder atoms having an isotopic distribution for a chemical element in proportional amounts to those usually found in nature).
  • atoms in at least three positions of the compound independently have an isotopic distribution for a chemical element in proportional amounts different to those usually found in nature (remainder atoms having an isotopic distribution for a chemical element in proportional amounts to those usually found in nature). In some embodiments, atoms in at least four positions of the compound independently have an isotopic distribution for a chemical element in proportional amounts different to those usually found in nature (remainder atoms having an isotopic distribution for a chemical element in proportional amounts to those usually found in nature).
  • atoms in at least five positions of the compound independently have an isotopic distribution for a chemical element in proportional amounts different to those usually found in nature (remainder atoms having an isotopic distribution for a chemical element in proportional amounts to those usually found in nature). In some embodiments, atoms in at least six positions of the compound independently have an isotopic distribution for a chemical element in proportional amounts different to those usually found in nature (remainder atoms having an isotopic distribution for a chemical element in proportional amounts to those usually found in nature).
  • Certain compounds for example those having incorporated radioactive isotopes such as 3 H and 14 C, are also useful in drug or substrate tissue distribution assays.
  • Tritium ( 3 H) and carbon-14 ( 14 C) isotopes are particularly preferred for their ease of preparation and detectability.
  • Compounds with isotopes such as deuterium ( 2 H) in proportional amounts greater than usually found in nature may afford certain therapeutic advantages resulting from greater metabolic stability, such as, for example, increased in vivo half-life or reduced dosage requirements.
  • Isotopically-labeled compounds can generally be prepared by performing procedures routinely practiced in the chemical art.
  • isotopic variant means a compound that contains an unnatural proportion of an isotope at one or more of the atoms that constitute such a compound.
  • an “isotopic variant” of a compound contains unnatural proportions of one or more isotopes, including, but not limited to, protium ( 1 H), deuterium ( 2 H), tritium ( 3 H), carbon-11 ( n C), carbon-12 ( 12 C), carbon-13 ( 13 C), carbon-14 ( 14 C), nitrogen- 13 ( 13 N), nitrogen- 14 ( 14 N), nitrogen- 15 ( 15 N), oxygen- 14 ( 14 O), oxygen- 15 ( 15 O), oxygen- 16 ( 16 O), oxygen- 17 ( 17 O), oxygen- 18 ( 18 O), fluorine- 17 ( 17 F), fluorine- 18 ( 18 F), phosphorus-31 ( 31 P), phosphorus-32 ( 32 P), phosphorus-33 ( 33 P), sulfur-32 ( 32 S), sulfur-33 ( 33 S), sulfur-34 ( 34 S), sulfur-35 ( 35 S
  • an “isotopic variant” of a compound is in a stable form, that is, non-radioactive.
  • an “isotopic variant” of a compound contains unnatural proportions of one or more isotopes, including, but not limited to, hydrogen ( X H), deuterium ( 2 H), carbon-12 ( 12 C), carbon-13 ( 13 C), nitrogen-14 ( 14 N), nitrogen-15 ( 15 N), oxygen- 16 ( 16 O), oxygen- 17 ( 17 O), and oxygen- 18 ( 18 O).
  • an “isotopic variant” of a compound is in an unstable form, that is, radioactive.
  • an “isotopic variant” of a compound of the disclosure contains unnatural proportions of one or more isotopes, including, but not limited to, tritium f'H), carbon-11 ( n C), carbon-14 ( 14 C), nitrogen-13 ( 13 N), oxygen-14 ( 14 O), and oxygen-15 ( 15 O).
  • any hydrogen can include 2 H as the major isotopic form, as example, or any carbon include be 13 C as the major isotopic form, as example, or any nitrogen can include 15 N as the major isotopic form, as example, and any oxygen can include 18 O as the major isotopic form, as example.
  • an “isotopic variant” of a compound contains an unnatural proportion of deuterium ( 2 H).
  • a position designated as having deuterium typically has a minimum isotopic enrichment factor of, in certain embodiments, at least 3500 (52.5% deuterium incorporation), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation) at each designated deuterium position.
  • the present disclosure further provides synthetic methods for incorporating radioisotopes into compounds of the disclosure. Synthetic methods for incorporating radioisotopes into organic compounds are well known in the art, and an ordinary skill in the art will readily recognize the methods applicable for the compounds of disclosure.
  • the present disclosure also relates to a method for the prevention or treatment of a disease or disorder mentioned herein comprising administering to a subject a compound of Formula I (e.g., a therapeutically effective amount of a compound of Formula I), or a pharmaceutically acceptable salt thereof.
  • a compound of Formula I e.g., a therapeutically effective amount of a compound of Formula I
  • a pharmaceutically acceptable salt thereof e.g., a pharmaceutically acceptable salt thereof.
  • the compound is administered in an amount of between about 1 mg and about 1000 mg per day, for example, between about 5 mg and about 500 mg per day, about 25 mg and about 400 mg per day, or about 50 mg and about 200 mg per day.
  • the word “between” is used to describe a numerical range, it is to be understood that the end points of the indicated range are explicitly included in the range. For example: if a temperature range is described to be between 40 °C and 80 °C, this means that the end points 40 °C and 80 °C are included in the range; or if a variable is defined as being an integer between 1 and 4, this means that the variable is the integer 1, 2, 3, or 4.
  • the term “about” (or alternatively the term “around”) placed before a numerical value “X” refers in the current application to an interval extending from X minus 10% of X to X plus 10% of X, and preferably to an interval extending from X minus 5% of X to X plus 5% of X.
  • the term “about” placed before a temperature “Y” refers in the current application to an interval extending from the temperature Y minus 10 °C to Y plus 10 °C, and preferably to an interval extending from Y minus 5 °C to Y plus 5 °C.
  • Example diseases or disorders where calcium T channels are involved include, but are not limited to:
  • epilepsy e.g. absence epilepsy, childhood absence and other forms of idiopathic generalized epilepsies, epileptic encephalopathy with continuous spike-and-wave during sleep, and temporal lobe epilepsy
  • pain e.g., inflammatory pain, neuropathic pain, peripheral pain, and chronic pain associated with peripheral axonal injury
  • neurological diseases and disorders e.g., essential tremors, Parkinson’s disease, schizophrenia, depression, anxiety, psychosis, neurodegenerative disorders, autism, and drug addiction
  • cardiovascular diseases and disorders e.g., hypertension, cardiac arrhythmias, atrial fibrillation, congenital heart failure, and heart block
  • the disease or disorder is selected from epilepsy, neurological disease and disorders, and pain. In some embodiments, the disease or disorder is epilepsy or pain. In some embodiments, the disease or disorder is neurological disease and disorders
  • the disease or disorder is epilepsy.
  • the epilepsy is selected from epileptic encephalopathy with continuous spike-and-wave during sleep (CSWS), and childhood absence epilepsy.
  • the epilepsy is epileptic encephalopathy with continuous spike-and-wave during sleep (CSWS).
  • the epilepsy is childhood absence epilepsy.
  • epilepsy describes recurrent unprovoked seizures wherein the term “seizure” refers to an excessive and/or hypersynchronous electrical neuronal activity.
  • Different types of “epilepsy” can be found, for example, Berg et al., Epilepsia, 2010; 51(4): 676-685, the disclosure of which is incorporated herein by reference in its entirety.
  • the term “epilepsy” as used herein preferably refers to absence epilepsy, childhood absence and other forms of idiopathic generalized epilepsies, temporal lobe epilepsy.
  • pain preferably refers to inflammatory pain, neuropathic pain, peripheral pain, or chronic pain associated with peripheral axonal injury.
  • neurodegenerative disorders preferably refers to essential tremors, Parkinson’s disease, schizophrenia, depression, anxiety, psychosis, neurodegenerative disorders, autism, or drug addiction.
  • the neurological diseases and disorders is essential tremor.
  • cardiac diseases and disorders preferably refers to hypertension, cardiac arrhythmias, atrial fibrillation, congenital heart failure, or heart block.
  • the compounds described herein are also useful in a method of reducing the concentration of calcium in a neuronal cell, and wherein said reduction in calcium is achieved by blocking the calcium T-channel present in such neuronal cell.
  • the compounds provide herein are also useful in a method of decreasing burst firing discharges in a neuronal cell and wherein said decrease of burst firing is achieved by blocking the calcium T-channel.
  • the method provided herein comprise administering a compound provided herein (i.e., a compound of any of Formulas I-V), or a pharmaceutically acceptable salt thereof.
  • the compounds provided herein may be metabolized by one or more cytochrome P450 isoforms.
  • cytochrome P450 isoforms in a subject include, but are not limited to, CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2A13, CYP2B6, CYP2C8, CYP2C9, CYP2C18, CYP2C19, CYP2D6, CYP2E1, CYP2G1, CYP2J2, CYP2R1, CYP2S1, CYP3A4, CYP3A5, CYP3A5P1, CYP3A5P2, CYP3A7, CYP4A11, CYP4B1, CYP4F2, CYP4F3, CYP4F8, CYP4F1, CYP4F12, CYP4X1, CYP1A1, CYP1A2, CYP1B1, CY
  • the term “contacting” refers to the bringing together of indicated moieties in an in vitro system or an in vivo system.
  • the term “patient” or “subject” used interchangeably refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, and most preferably humans.
  • the phrase “therapeutically effective amount” refers to the amount of active compound or pharmaceutical agent such as an amount of any of the solid forms or salts thereof as disclosed herein that elicits the biological or medicinal response in a tissue, system, animal, subject, or human that is being sought by a researcher, veterinarian, medical doctor or other clinician.
  • An appropriate “effective” amount in any individual case may be determined using techniques known to a person skilled in the art.
  • phrases “pharmaceutically acceptable” is used herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, immunogenicity or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • the phrase “pharmaceutically acceptable carrier or excipient” refers to a pharmaceutically-acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, solvent, or encapsulating material. Excipients or carriers are generally safe, non-toxic and neither biologically nor otherwise undesirable and include excipients or carriers that are acceptable for veterinary use as well as human pharmaceutical use. In some embodiments, each component is “pharmaceutically acceptable” as defined herein.
  • treating refers to inhibiting the disease; for example, inhibiting a disease, condition or disorder in a subject who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., arresting further development of the pathology and/or symptomatology) or ameliorating the disease; for example, ameliorating a disease, condition or disorder in a subject who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., reversing the pathology and/or symptomatology) such as decreasing the severity of disease.
  • the compounds of the disclosure are useful in preventing or reducing the risk of developing any of the diseases referred to herein; e.g., preventing or reducing the risk of developing a disease, condition or disorder in a subject who may be predisposed to the disease, condition or disorder but does not yet experience or display the pathology or symptomatology of the disease.
  • certain features of the disclosure which are, for clarity, described in the context of separate embodiments, can also be provided in combination in a single embodiment (while the embodiments are intended to be combined as if written in multiply dependent form).
  • various features of the disclosure which are, for brevity, described in the context of a single embodiment, can also be provided separately or in any suitable subcombination.
  • the compounds provided herein can also be used, for example, in one or more methods and/or uses described herein in combination with one or more additional therapeutic agents.
  • the additional therapeutic agent is an antiepileptic therapeutic agent, or a pharmaceutically acceptable salt thereof. Examples of additional therapeutic agents useful in combination with the compounds provided herein can be found, for example, in U.S. Patent No.: 11,213,517, the disclosure of which is incorporated herein by reference in its entirety.
  • the additional therapeutic agent is selected from 6-(2,3- dichlorophenyl)-l,2,4-triazine-3,5-diamine, (S)-2-(2-oxopyrrolidin-l-yl)butanamide, and 2-propylpentanoic acid, or a pharmaceutically acceptable salt of any of the aforementioned.
  • the additional therapeutic agent is 6-(2,3- dichlorophenyl)-l,2,4-triazine-3,5-diamine, or a pharmaceutically acceptable salt thereof.
  • the additional therapeutic agent is (S)-2-(2-oxopyrrolidin-l- yl)butanamide, or a pharmaceutically acceptable salt thereof.
  • the additional therapeutic agent is 2-propylpentanoic acid, or a pharmaceutically acceptable salt thereof.
  • the compounds provided herein and the additional therapeutic agents are comprised in a single pharmaceutical composition. In some embodiments, the compound provided herein and the additional therapeutic agents are comprised in separated pharmaceutical compositions (e.g., two or more pharmaceutical compositions). In some embodiments, the compounds provided herein and the additional therapeutic agents are administered simultaneously. In some embodiments, the compound provided herein and the additional therapeutic agents are administered sequentially.
  • the combination exhibits synergistic effect. In some embodiments, the combination exhibits additive effect.
  • compositions can be effected in a manner which will be familiar to any person skilled in the art (see e.g., Remington, The Science and Practice of Pharmacy 21st Edition (2005), Part 5, “Pharmaceutical Manufacturing” [published by Lippincott Williams & Wilkins]) by bringing the described compounds of formula (I), or their pharmaceutically acceptable salts, optionally in combination with other therapeutically valuable substances, into a galenical administration form together with suitable, non-toxic, inert, therapeutically compatible solid or liquid carrier materials and, if desired, usual pharmaceutical adjuvants.
  • the compounds provided herein, and pharmaceutically acceptable salts thereof, can be used as medicaments, e.g., in the form of pharmaceutical compositions for enteral (e.g., oral) or parenteral administration, including topical application or inhalation.
  • LC-MS Analytical UPLC on a Agilent Zorbax RRHD SB-Aq (2.1x50mm, 1 ,8um); detection at 210 nm and MS; Gradient of water/ 0.04% TFA (A) and MeCN (B).
  • the eluent flow rate was 0.8 mL/min and the characteristics of the eluting mixture proportion in function of the time t from start of the elution are summarized in the table below (a linear gradient being used between two consecutive time points):
  • Preparative HPLC/MS purifications are performed on a Gilson HPLC system, equipped with a Gilson 215 autosampler, Gilson 333/334 pumps, Finnigan AQA MS detector system, and a Dionex UV detector, using a Waters Xbridge Cl 8 or an Waters Atlantis column, with a linear gradient of water/formic acid 0.02% (A) and MeCN (B) (acidic conditions) or water/ammonia 0.02% (A) and MeCN (B) (basic conditions).
  • the amine 5’ undergoes an amide coupling with the known acid 6’ (O. Bezemjon et al., J. Med Chem 2017,60, 9769-9789) using an activating agent such as HATU in presence of a base such as DIPEA to give amide 7’.
  • the bromopyrazole 7’ is converted into the deuterated analog 1’ using a catalyst such as Pd(OH)2 in a presence of a base such as NaOAc in a solvent such as EtOAc or Pd/C in a presence of a base such as NEt3 in a deuterated solvent like D3COD under a D2- atmosphere.
  • Nitropyrazole 21’ is alkylated with the bromide 3’ in the presence of a base such as K 2 CO 3 to give the nitroderivative 22’.
  • a base such as K 2 CO 3
  • Deuteration and concomitant nitroreduction of the dibromoderivative 22’ using a palladium catalyst in the presence of a base under D 2 -atmosphere affords deuterated amine 23’.
  • a final amide coupling between amine 23’ and the acid 6’ using an activating agent such as HATU in the presence of a base such as DIPEA yields compound 20’.
  • 2-(4-(l-(trifluoromethyl)cyclopropyl)phenyl)acetamide) is prepared as presented in Scheme 5.
  • Alcohol 13’ is activated e.g. as a mesylate and used to alkylate nitropyrazole 21’ in the presence of a base to give compound 25’.
  • Deuteration and concomitant nitroreduction of the dibromoderivative 25’ yields the amine 26’ which undergoes an amide coupling with acid 6’ in the presence of an activating agent such as HATU and a base such as DIPEA to give the final compound 24’.
  • an activating agent such as HATU
  • DIPEA a base
  • Step 3 N-(4-bromo-l-((5-cyanopyridin-2-yl)methyl)-lH-pyrazol-3-yl)-2-(4-(l-)
  • Step 4 N-( I -( (5-cyariopyridin-2-yl)methyl)-lH-pyrazol-3-yl-4-d)-2-(4-( 1 - (trifluoromethyl)cyclopropyl)phenyl)acetamide (1 ’)
  • Step 4 N-( 1 -( (5-cyanopyridin-2-yl)methyl)-lH-pyrazol-3-yl-5-d)-2-(4-( 1 - ( trifluoromethyl)cyclopropyl)phenyl)acetamide (8’)
  • Step 2 N-( I -( (5-cyanopyridin-2-yl)methyl)-lH-pyrazol-3-yl)-2-(4-( I - (trifluoromethyl)cyclopropyl)phenyl)acetamide-2,2-d2 (18’)
  • the mixture was kept at 4 °C for 18 hours.
  • the mixture was diluted with EtOAc (25 mL) and washed with 0.1M aq. HC1 soln. (25 mL), sat. aq. NaHCOs (25 mL), sat. aq. NaCl soln. (25 mL), dried over MgSO4 and concentrated in vacuo.
  • the residue was purified by prep. HPLC (column: Water X- Bridge, 30x75 mm, 10 um, UV/MS, basic conditions). The fractions containing product were concentrated in vacuo.
  • the residue was diluted with sat. aq. NaHCOs soln. (10 mL) and extracted with DCM (3 x 10 mL).
  • Example 2 In vitro Methods - Measurement of calcium channel flux by means of FLIPR assays.
  • HEK293 cells recombinantly expressing either voltage-dependent T-type calcium channel subunit alpha- 1G (Cav3.2) or voltage-dependent L-type calcium channel subunit alpha-lC (Cavl.2) are assayed for calcium flux using the calcium indicator dye Calcium 6 (Molecular Devices) and FLIPR technology (Fluorometric Imaging Plate Reader, Molecular Devices) (Xie X, Van Deusen AL, Vitko I, Babu DA, Davies LA, Huynh N, Cheng H, Yang N, Barrett PQ, Perez-Reyes E. Validation of high throughput screening assays against three subtypes of Ca(v)3 T-type channels using molecular and pharmacologic approaches.
  • the HEK293 cells recombinantly expressing Cav3.2 are maintained in DMEM growth medium Gibco) supplemented with 10 % Fetal Bovine Serum (FBS), 100 U/ml penicilin (Gibco), 100 pg/ml streptomycin (Gibco) and 1 mg/ml G418 (Gibco).
  • HEK293 cells recombinantly expressing Cavl .2 are maintained in MEM (Gibco), 10% FBS (Gibco); 2mM L-Glutamine (Gibco); 1% Pen/strep; 400 pg/ml G418; 10 pg/ml Zeocin.
  • Cells are washed once with PBS, then dissociated in 0.25 % trypsin/EDTA (Gibco) and seeded into PureCoat Amine coated 384-well black (Corning), clear bottom plates at a density of 20,000 cells/well, in culture medium for the HEK-hCav3.2 cell line, and without antibiotics forthe HEK-hCavl.2 cell line. The seeded plates are incubated overnight at 37°C, 5% CO2.
  • HBSS IX (137 mM NaCl; 5.4 mM KCl; 0.25 mM Na2HPC>4; 1.3 mM CaCl 2 ; 0.4 mM MgSCU; 0.5 mM MgCh; 0.4 mM KH2PO4), 0.375 g/L NaHCCh, 20 mM Hepes,l% FBS (Gibco), pH 7.4.
  • HBSS IX 137 mM NaCl; 5.4 mM KCl; 0.25 mM Na2HPC>4
  • CaCl 2 0.4 mM MgSCU; 0.5 mM MgCh; 0.4 mM KH2PO4
  • 0.375 g/L NaHCCh 20 mM Hepes,l% FBS (Gibco), pH 7.4.
  • the cells are loaded in presence of Probenecid (AATBioquest; 2.5 mM final concentration in loading buffer) for 1 hour at 37°C. Then, the loading buffer is discarded, and cells are kept in 50 pl/well of assay buffer (HBSS IX; 0.375 g/L NaHCCL; 20 mM Hepes; 1 % FBS; pH 7.4) for 30 min at RT in the dark.
  • AATBioquest Probenecid
  • HBSS IX 0.375 g/L NaHCCL
  • 20 mM Hepes 1 % FBS; pH 7.4
  • test compounds are prepared to a concentration of 10 mM in DMSO.
  • TEAC buffer 100 mM tetraethylammonium chloride; 20 mM Hepes; 2.5 mM CaCh; 5 mM KC1; 1 mM MgCE; 1 % FBS; pH 7.2
  • assay buffer HBSS IX; 0.375 g/L NaHCCh; 20 mM Hepes; 1 % FBS; pH 7.4
  • Test compounds are added to the cells to give a 3-fold dilution range from 10 pM to 0 05 nM
  • the compounds are incubated with the cells for 3 minutes and Ca 2+ entry is stimulated by adding either CaCh to a final concentration of 10 mM (Cav3.2 assay) or by adding KC1 to a final concentration of 75 mM (Cavl.2 assay).
  • the kinetics of fluorescence increase are recorded for every well and the area under the fluorescence trace for every compound concentration is used to generate inhibition curves using non-linear regression sigmoidal concentration-response curve analysis with in-house software.
  • IC50 values are calculated and represent the compound concentration required to inhibit 50% of the signal that is obtained in the presence of vehicle instead of test compound.
  • Antagonistic activities (IC50 values) have been measured for the for the Cav3.1- and the Cav3.3 -channel.
  • the metabolism of Compound 2 was investigated using liver microsomes and hepatocytes of man and a set of animal species used or considered to be used in preclinical safety testing.
  • the in vitro metabolic profile of Compound 2 with human liver preparations was characterized by the formation of five metabolites, i.e., M1-M5. It was found that Compound 2 undergoes three primary metabolic pathways: oxidative dealkylation of the pyrazole ring to form Ml, hydrolysis of the amide bond to form M2, and hydroxylation to yield M29.
  • M1-M5 five metabolites
  • Ml was the product of aP450/FMO-dependent reaction and undergoes further conjugation with pentose to yield the humanspecific metabolite, M3. Hydrolysis to M2 was shown to be catalyzed by microsomal enzymes other than cytochrome P450s.
  • M5 is a cysteine conjugate of 3-aminopyrazole.
  • Compound 2 was hydrolyzed to M2 in plasma of rat, monkey and mouse whereas no degradation was observed in human and rabbit plasma. M2 was also formed in blood from human, rat and monkey, but not in blood from rabbit and mouse.
  • Compound 2 is the 14 C-labelled analogue of Compound 1 bearing the radiolabel in the pyrazole moiety of the molecule.
  • the 14 C labeled Compound 2 was used as a tool compound.
  • the deuterated compounds can also be used instead of Compound 2 for any of the assays described herein.
  • the objective of this in vitro study was the determination and comparison of Compound 2 metabolic profiles using liver microsomes, hepatocytes, blood, and plasma, from a number of animal species envisaged for toxicity testing, as well as of man.
  • the number and proportion of metabolites generated following incubation of the 14 C-labelled analogue, Compound 2 at a single concentration of 10 pM with liver preparations from CD-I mouse, Wistar rat, NZW rabbit, Beagle dog, cynomolgus monkey, and man were determined using HPLC coupled with 14C-radiodetection.
  • the stability in plasma and blood of all species used in the safety evaluation was investigated as Compound 2 hydrolyzed to metabolite M2. Data on plasma and blood stability were generated in support of the plasma protein binding and blood partitioning studies.
  • Glucose-6-phosphate dehydrogenase was supplied by Roche Diagnostics (Mannheim, Germany).
  • the liquid scintillation cocktail for HPLC analysis, Optiflow Safe 2 was purchased from Berthold Technologies GmbH (Regensdorf, Switzerland). Leibovitz's L-15 and William’s E media were supplied from Life Technologies (Basel, Switzerland).
  • the HPLC/MS system for the recording of metabolic profiles and mass spectra consisted of two Shimadzu pumps LC-30AD (Shimadzu, Reinach, Switzerland) equipped with a Shimadzu membrane degasser DGU-30A5, a Shimadzu diode array detector SPD- M20A, a Shimadzu column oven CTO-20A, and a Shimadzu autosampler model SIL- 20 AC. Radio detection was performed by a Berthold radioflow detector LB513 with a 200 pL liquid cell Z-200-6M, a LB5036 pump for supplementing liquid scintillation cocktail at 3 mL/min (Berthold AG, Regensdorf, Switzerland).
  • Detection of mass data was performed by a LTQ XL linear ion trap mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). The acquisition and analysis of radiochemical and mass data were done using the RadioStar (version 5.0.12.4, Berthold AG, Regensdorf, Switzerland) and Xcalibur software packages (version 2.2 SP1.48, Thermo Fisher Scientific, Waltham, MA, USA). As used herein, the disclosed m/z values refer to the singly protonated [M+H] + . The following MS equipment and parameters were used:
  • the NADPH-regenerating system used for the liver microsomal incubations was prepared as a 10-fold concentrated stock solution and kept at -20 °C. It consisted of 11 mM NADP, 100 mM glucose-6-phosphate and 50 mM magnesium chloride in 0.1 M phosphate buffer (pH 7.4). 20 lU/mL of glucose-6-phosphate dehydrogenase was added before use.
  • Cryopreserved human hepatocytes (batch 3) were provided by Celsis (Neuss, Germany), while NZW rabbit and CD-I mouse cells were provided by Biopredic International (Rennes, France).
  • Hepatocytes from Wistar rat were prepared at Actelion Pharmaceutical Ltd following the standard two-step collagenase perfusion method (see e.g.. Seglen & Fossa, Exp. Cell Res. 1978, 116: 199-206).
  • Incubations of Compound 2 with liver microsomes of all species were performed at a single substrate concentration of 10 pM in 2 mL amber reaction vials.
  • a 2.0 pL-aliquot of the Compound 2 stock solution was added to 100 mM phosphate buffer (pH 7.4) containing the liver microsomes at a protein concentration of 1 mg/mL for human, rabbit, rat, monkey and mouse liver microsomes, and at 3 mg/mL for dog liver microsomes.
  • the mixture was incubated at 37 °C in an Eppendorf thermomixer and agitation at 650 rpm.
  • the organic solvent concentration in all incubations was kept ⁇ 1 % (v/v).
  • the reaction was initiated by addition of 20 pL of prewarmed NADPH-regenerating system containing the glucose-6-phosphate dehydrogenase and terminated either immediately after the start of the reaction (control) or after 60 minutes, by addition of 200 pL of ice-cold acetonitrile. Samples were centrifuged at 20'800 g and 10 °C for 5 min. Prior to HPLC analysis, a 100 pL-aliquot of the supernatant was mixed with 300 pL mobile phase A in order to mimic initial HPLC conditions. Control experiments were performed in the absence of either the NADPH- regenerating system or the liver microsomes under otherwise identical conditions. Tn these controls, the volumes of both co-factors were replaced by 100 mM phosphate buffer (pH 7.4).
  • Freshly prepared hepatocytes were cultured in William’s E medium supplemented with 10 % fetal bovine serum and 4 pg/mL bovine insulin. Cryopreserved cells were thawed and the supplied medium was replaced by culture medium. Incubations with Compound 2 were performed using a culture medium additionally fortified with 0.48 pg/mL hydrocortisone and 400 pM L-glutamine. Neither the culture nor the incubation media contained phenol red or antibiotics.
  • Freshly isolated and collagenase-perfused rat liver was kept in Leibovitz's medium and was mechanically dissociated using sterile pipette tips.
  • the cell suspension was transferred through a nylon mesh cell strainer with a pore size of 70 pm into a sterile 50 mb centrifugation tube.
  • the cell suspension was centrifuged at 50 g and 4 °C for 4 min, the cell pellet re-suspended in Leibovitz’s medium, followed by purification on a Percoll cushion (15%) and another centrifugation step at 50 g and 4 °C for 4 min. After centrifugation, cells were resuspended in 1 mL culture medium.
  • Vi-Cell viability analyzer (Beckman Coulter, Nyon, Switzerland) for the determination of cell number and initial cell viability, by a trypan blue dye exclusion test. Viabilities are summarized in Table 2.
  • the cell suspension was then adjusted with culture medium at a nominal density of 5 x 105 viable cells/mL. 400 pL-aliquots of this suspension were dispensed into collagen-coated 24-well plates and incubated at 37 °C for a period of about 3 h in a humidified atmosphere containing 5 % CO2.
  • the medium was removed from each well and replaced by 200 pL of pre-warmed (37 °C) incubation medium containing Compound 2 at a final concentration of 10 pM.
  • Triplicate wells were sampled after 0, 4 and 24 h of incubation by addition of 200 pL of ice-cold acetonitrile. The entire well content was transferred into 2 mL amber reaction vials. Samples were stored frozen at - 20 °C pending analysis. Prior to HPLC analysis, samples were thawed at 37 °C, vortex- mixed and centrifuged at 20'800 g and 10 °C for 5 min. A 100 pL-aliquot of the supernatant was mixed with 300 pL mobile phase A in order to mimic initial HPLC conditions and submitted to HPLC analysis.
  • Cryopreserved hepatocytes from NZW rabbit were thawed at 37 °C, and purified on a Percoll cushion (15%) at 50 g for 4 min at 4 °C.
  • the resulting cell pellet was resuspended in 1 mL culture medium and cell viabilities determined as described above.
  • the cell suspension was adjusted with incubation medium at a nominal density of 1 x 10 6 viable cells/mL. 500 pL-aliquots of this cell suspension were dispensed into 2 mL amber reaction vials and placed in an Eppendorf thermomixer at 37 °C and 850 rpm to keep the cells in suspension.
  • Plasma and blood of cynomolgus monkey, Wistar rat, CD-I mouse, New Zealand White rabbit and man were fortified with Compound 2 at a final concentration of 10 pM in a total reaction volume of 1 mL. After an incubation time of 0.5, 1 and 2 h for blood, or 2, 4 and 6 h for plasma, 200 pL-aliquots of the incubation mixture were mixed with 600 pL of a 8:2 (v/v) mixture of acetonitrile and methanol to lyse blood cells and precipitate proteins.
  • Recoveries were determined as the ratio of total radioactivity before and after centrifugation of the quenched incubation mixtures. For this purpose, triplicate 10 pL aliquots of each incubation were mixed with 4 mL of IRGA SafePlus liquid scintillation cocktail and submitted for liquid scintillation counting using a Tricarb 2300 TR liquid scintillation analyzer (Perkin Elmer, Zurich, Switzerland) with luminescence correction and on-line quenching correction by means of an internal standard. Results of the recovery determinations are summarized in FIG. 1. Mean recoveries were in excess of 97% and 84% for liver microsomal and hepatocyte incubations, respectively.
  • M4 is a secondary metabolite and the product of hydroxylation in the pyrazole amide moiety followed by glucuronidation.
  • M29 the aglycon of M4, (i.e., the primary hydroxylation product of the pyrazole moiety) was only detected after treatment with ⁇ -glucuronidase.
  • Mass spectrometry data indicate that M29 was not present in incubations with Compound 2 with liver microsomes or hepatocytes. The exact chemical structure of M4 and its precursor is yet unknown. Conjugation of Ml with a pentose gives the phase II metabolite M3.
  • M5 is a cysteine conjugate of 3-aminopyrazole.
  • Compound 2 was incubated at a single substrate concentration of 10 pM for up to 60 minutes with liver microsomes of CD-I mouse, Wistar rat, NZW rabbit, Beagle dog, cynomolgus monkey and man at microsomal protein concentrations of 1 or 3 mg/mL. Control experiments in the absence of liver microsomes or the NADPH-regenerating system were performed in order to demonstrate that metabolite formation was indeed P450/FMO-dependent.
  • FIG. 3 gives an overview on the number of metabolites formed and their individual relative contributions. The radiochromatograms of these incubations including controls are presented in FIGs. 4-11.
  • FIG. 12 gives an overview on the number of metabolites formed in the different incubations together with their individual relative contributions.
  • the respective radiochromatograms including the control without cells are presented in FIGs. 13-22.
  • the metabolism of Compound 2 with human hepatocytes was investigated using four different batches of human liver cells (FIGs. 13-16). Up to five products, designated M1-M5, were observed. All five metabolites were seen following 24 h-incubation of Compound 2 with batches 3 and 4, none individually exceeding 21 % and with an overall turnover of 32% and 52%, respectively. Only Ml was observed with human hepatocyte batches 1 , 2 and 3 after an incubation time of 4 h. Metabolites M2 and M3 were detected in batch 1 after 24 h-incubation, accounting for 9.2% and 3.1%, respectively. Metabolites Ml and M2 were detected in batch 2 after 24 h-incubation representing 3.1% and 10%, respectively. Control experiments in the absence of liver cells confirmed that all five products were indeed Compound 2 metabolites (FIG. 22).
  • Compound 2 yielded five metabolites with rat hepatocytes (FIG. 17) after 24 h incubation and a turnover of 88%.
  • dog and mouse hepatocytes yielded metabolites Ml and M2, accounting for 4.2% and 8.5% in dog liver cells, respectively, while 12% and 8.0% were observed in mouse liver cells.
  • Rabbit hepatocytes (FIG. 21) resulted in the formation of M2 accounting for 4.0% after 6 h of incubation.
  • Compound 2 was incubated with plasma of Wistar rat, CD-I mouse, NZW rabbit, cynomolgus monkey and man at 37 °C for 2, 4 and 6 hours, as well as with blood of the same species for 0.5, 1 and 2 hours.
  • the rate constant k of the hydrolysis to M2 was determined from the slope of log concentration versus time plot.
  • Metabolic profiles of microsomal and hepatocytes incubations were compared to assign metabolites to phase I or phase II metabolism.
  • the primary metabolite Ml observed in liver microsomes from all species was also detected in hepatocyte incubations of human, dog, monkey and mouse. Since its formation was dependent on NADPH, it is most likely a product of a P450/FMO-catalyzed reaction.
  • the hydrolysis product M2 observed in hepatocytes of all species was only observed in microsomes of rat and mouse. Its formation in the absence of NADPH suggests a P450-independent hydrolysis.
  • Metabolites M3 and M4 are products of phase II metabolism and were only present in hepatocyte incubations.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Endocrinology (AREA)
  • Rheumatology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Diabetes (AREA)
  • Hematology (AREA)
  • Obesity (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 des composés deutérés et leur utilisation en tant que bloqueurs des canaux calciques de type T dans le traitement ou la prévention de diverses maladies ou troubles associés aux canaux calciques T.
PCT/US2023/068152 2022-06-10 2023-06-08 Composés deutérés WO2023240205A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US202263350887P 2022-06-10 2022-06-10
US63/350,887 2022-06-10
US202363458323P 2023-04-10 2023-04-10
US63/458,323 2023-04-10

Publications (1)

Publication Number Publication Date
WO2023240205A1 true WO2023240205A1 (fr) 2023-12-14

Family

ID=87136681

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2023/068152 WO2023240205A1 (fr) 2022-06-10 2023-06-08 Composés deutérés

Country Status (1)

Country Link
WO (1) WO2023240205A1 (fr)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015186056A1 (fr) 2014-06-03 2015-12-10 Actelion Pharmaceuticals Ltd Composés de pyrazole et leur utilisation en tant que bloqueurs des canaux calciques de type t
US10246426B2 (en) 2014-09-15 2019-04-02 Idorsia Pharmaceuticals Ltd Triazole compounds as T-type calcium channel blockers
WO2020203610A1 (fr) 2019-03-29 2020-10-08 日本ケミファ株式会社 Utilisation d'un bloqueur des canaux calciques de type t servant au traitement de la polyarthrite rhumatoïde
US10899695B2 (en) 2017-02-06 2021-01-26 Idorsia Pharmaceuticals Ltd Process for the synthesis of 1-aryl-1-trifluoromethylcyclopropanes
US11059803B2 (en) 2017-07-05 2021-07-13 Idorsia Pharmaceuticals Ltd Crystalline form of N-[1-(5-cyano-pyridin-2- ylmethyl)-1H-pyrazol-3-yl]-2-[4-(1-trifluoromethyl-cyclopropyl)-phenyl]-acetamide
US11213517B2 (en) 2016-12-16 2022-01-04 Idorsia Pharmaceuticals Ltd Pharmaceutical combination comprising a T-type calcium channel blocker

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015186056A1 (fr) 2014-06-03 2015-12-10 Actelion Pharmaceuticals Ltd Composés de pyrazole et leur utilisation en tant que bloqueurs des canaux calciques de type t
US9932314B2 (en) 2014-06-03 2018-04-03 Idorsia Pharmaceuticals Ltd Pyrazole compounds and their use as T-type calcium channel blockers
US10065929B2 (en) 2014-06-03 2018-09-04 Idorsia Pharmaceuticals Ltd Pyrazole compounds and their use as T-type calcium channel blockers
US10246426B2 (en) 2014-09-15 2019-04-02 Idorsia Pharmaceuticals Ltd Triazole compounds as T-type calcium channel blockers
US11213517B2 (en) 2016-12-16 2022-01-04 Idorsia Pharmaceuticals Ltd Pharmaceutical combination comprising a T-type calcium channel blocker
US10899695B2 (en) 2017-02-06 2021-01-26 Idorsia Pharmaceuticals Ltd Process for the synthesis of 1-aryl-1-trifluoromethylcyclopropanes
US11059803B2 (en) 2017-07-05 2021-07-13 Idorsia Pharmaceuticals Ltd Crystalline form of N-[1-(5-cyano-pyridin-2- ylmethyl)-1H-pyrazol-3-yl]-2-[4-(1-trifluoromethyl-cyclopropyl)-phenyl]-acetamide
WO2020203610A1 (fr) 2019-03-29 2020-10-08 日本ケミファ株式会社 Utilisation d'un bloqueur des canaux calciques de type t servant au traitement de la polyarthrite rhumatoïde

Non-Patent Citations (11)

* Cited by examiner, † Cited by third party
Title
"Pharmaceutical Preformulation and Formulation", 2009, THE PHARMACEUTICAL PRESS AND THE AMERICAN PHARMACEUTICAL ASSOCIATION
"Remington, The Science and Practice of Pharmacy", 2005, LIPPINCOTT WILLIAMS & WILKINS: PHILADELPHIA, article "Pharmaceutical Manufacturing"
A. KEREKES, J. MED. CHEM., vol. 54, 2011, pages 201 - 210
ALAN F. THOMAS: "Deuterium Labeling in Organic Chemistry", 1971, APPLETON-CENTURY-CROFTS
BAHAR ET AL., J. PHARM. SCI, vol. 101, 2012, pages 3979 - 88
BERG ET AL., EPILEPSIA, vol. 51, no. 4, 2010, pages 676 - 685
BEZENÇON ET AL., J. MED. CHEM., vol. 60, 2017, pages 9769 - 9789
JAMES R. HANSON: "The Organic Chemistry of Isotopic Labelling", 2011, ROYAL SOCIETY OF CHEMISTRY
R. XU, J. LABEL COMPD. RADIOPHARM., vol. 58, 2015, pages 308 - 312
SEGLENFOSSA, EXP. CELL RES, vol. 116, 1978, pages 199 - 206
XIE XVAN DEUSEN ALVITKO IBABU DADAVIES LAHUYNH NCHENG HYANG NBARRETT PQPEREZ-REYES E: "Validation of high throughput screening assays against three subtypes of Ca(v)3 T-type channels using molecular and pharmacologic approaches", ASSAY AND DRUG DEVELOPMENT TECHNOLOGIES, vol. 5, no. 2, 2007, pages 7744 - 7765

Similar Documents

Publication Publication Date Title
JP5656408B2 (ja) 心臓の神経支配を画像化するためのリガンド
EP2300481B1 (fr) Nouveaux azabenzoxazoles substitués
Wang et al. [11C] GSK2126458 and [18F] GSK2126458, the first radiosynthesis of new potential PET agents for imaging of PI3K and mTOR in cancers
CN109641887B (zh) 可用作用于治疗癌症的突变idh1抑制剂的噻唑衍生物
EP2252605B1 (fr) Analogues marqués d'halogéno-benzamides en tant que produits radiopharmaceutiques multimodaux et leurs précurseurs
TWI518087B (zh) 咪唑并［１，２－ａ］吡啶－７－胺
US11534434B2 (en) Xanomeline derivatives and methods for treating neurological disorders
CA2835018A1 (fr) Derives de pentylenetetrazole
WO2023240205A1 (fr) Composés deutérés
US20220144817A1 (en) Compounds and methods of deuterated xanomeline for treating neurological disorders
Langer et al. Synthesis of fluorine-18-labeled ciprofloxacin for PET studies in humans
WO2023202568A1 (fr) Composé de sel d'ammonium quaternaire, son procédé de préparation et son utilisation
EP3743427B1 (fr) Nouveau procédé de préparation d'un composé d'imagerie
US10406126B2 (en) ALDH2 activator
Gao et al. [11C] Dimebon, radiosynthesis and lipophilicity of a new potential PET agent for imaging of Alzheimer’s disease and Huntington’s disease
Marchais-Oberwinkler et al. N-Oxide analogs of WAY-100635: new high affinity 5-HT1A receptor antagonists
JP7262141B2 (ja) シャペロン介在性オートファジー調節剤として有用な化合物
US12006302B2 (en) Tau PET imaging ligands
WO2024064178A1 (fr) Inhibiteurs d'hexahydro-2h-pyrido[2,1-a]isoquinoline vmat2 et procédés d'utilisation
EP4396162A2 (fr) Allyl tryptamines asymétriques
WO2023034645A2 (fr) Allyl tryptamines asymétriques
WO2024091523A1 (fr) Analogues de phényléthylamine à conformation restreinte
Liu et al. A carbon-11 labeled imidazo [1, 2-a] pyridine derivative as a new potential PET probe targeting PI3K/mTOR in cancer
KR20230077667A (ko) 신규 양성자방출단층촬영 방사성 화합물 및 이의 용도
WO2023049480A1 (fr) Phénylalkylamines substituées

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: 23738392

Country of ref document: EP

Kind code of ref document: A1