Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/08—Tripeptides
  • C07K5/0819—Tripeptides with the first amino acid being acidic
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P11/00—Drugs for disorders of the respiratory system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P11/00—Drugs for disorders of the respiratory system
  • A61P11/06—Antiasthmatics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P13/00—Drugs for disorders of the urinary system
  • A61P13/12—Drugs for disorders of the urinary system of the kidneys
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/06—Antipsoriatics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P19/00—Drugs for skeletal disorders
  • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P19/00—Drugs for skeletal disorders
  • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
  • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
  • A61P21/02—Muscle relaxants, e.g. for tetanus or cramps
• • 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
• • 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
  • A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
• • 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
  • A61P25/08—Antiepileptics; Anticonvulsants
• • 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
  • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
• • 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
  • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
  • A61P25/16—Anti-Parkinson drugs
• • 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
  • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P27/00—Drugs for disorders of the senses
  • A61P27/02—Ophthalmic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• • 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
  • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
  • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/04—Antibacterial agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/12—Antivirals
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/12—Antivirals
  • A61P31/14—Antivirals for RNA viruses
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/12—Antivirals
  • A61P31/14—Antivirals for RNA viruses
  • A61P31/18—Antivirals for RNA viruses for HIV
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/12—Antivirals
  • A61P31/20—Antivirals for DNA viruses
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
  • A61P35/02—Antineoplastic agents specific for leukemia
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
  • A61P35/04—Antineoplastic agents specific for metastasis
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
  • A61P37/02—Immunomodulators
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
  • A61P37/02—Immunomodulators
  • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
  • A61P7/06—Antianaemics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
  • A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
  • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C271/00—Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
  • C07C271/06—Esters of carbamic acids
  • C07C271/08—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
  • C07C271/10—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms
  • C07C271/22—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C317/00—Sulfones; Sulfoxides
  • C07C317/44—Sulfones; Sulfoxides having sulfone or sulfoxide groups and carboxyl groups bound to the same carbon skeleton
  • C07C317/48—Sulfones; Sulfoxides having sulfone or sulfoxide groups and carboxyl groups bound to the same carbon skeleton the carbon skeleton being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups
  • C07C317/50—Sulfones; Sulfoxides having sulfone or sulfoxide groups and carboxyl groups bound to the same carbon skeleton the carbon skeleton being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups at least one of the nitrogen atoms being part of any of the groups, X being a hetero atom, Y being any atom
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
  • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
  • C07D209/04—Indoles; Hydrogenated indoles
  • C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
  • C07D209/14—Radicals substituted by nitrogen atoms, not forming part of a nitro radical
  • C07D209/16—Tryptamines
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
  • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
  • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
  • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
  • C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
  • C07D213/81—Amides; Imides
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
  • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
  • C07D215/12—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
  • C07D295/22—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with hetero atoms directly attached to ring nitrogen atoms
  • C07D295/26—Sulfur atoms
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
  • C07F9/02—Phosphorus compounds
  • C07F9/28—Phosphorus compounds with one or more P—C bonds
  • C07F9/38—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)]
  • C07F9/40—Esters thereof
  • C07F9/4003—Esters thereof the acid moiety containing a substituent or a structure which is considered as characteristic
  • C07F9/4006—Esters of acyclic acids which can have further substituents on alkyl
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
  • C07F9/02—Phosphorus compounds
  • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
  • C07F9/6527—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07F9/6533—Six-membered rings
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/81—Protease inhibitors
  • C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
  • C07K14/8139—Cysteine protease (E.C. 3.4.22) inhibitors, e.g. cystatin
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
  • C07K5/0202—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-X-X-C(=0)-, X being an optionally substituted carbon atom or a heteroatom, e.g. beta-amino acids
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/06—Dipeptides
  • C07K5/06008—Dipeptides with the first amino acid being neutral
  • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
  • C07K5/06034—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/08—Tripeptides
  • C07K5/0802—Tripeptides with the first amino acid being neutral
  • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
  • C07K5/081—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing O or S as heteroatoms, e.g. Cys, Ser
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/08—Tripeptides
  • C07K5/0802—Tripeptides with the first amino acid being neutral
  • C07K5/0812—Tripeptides with the first amino acid being neutral and aromatic or cycloaliphatic
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/10—Tetrapeptides
  • C07K5/1002—Tetrapeptides with the first amino acid being neutral
  • C07K5/1005—Tetrapeptides with the first amino acid being neutral and aliphatic
  • C07K5/1013—Tetrapeptides with the first amino acid being neutral and aliphatic the side chain containing O or S as heteroatoms, e.g. Cys, Ser
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/005—Enzyme inhibitors
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C2602/00—Systems containing two condensed rings
  • C07C2602/02—Systems containing two condensed rings the rings having only two atoms in common
  • C07C2602/04—One of the condensed rings being a six-membered aromatic ring
  • C07C2602/08—One of the condensed rings being a six-membered aromatic ring the other ring being five-membered, e.g. indane
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/06—Dipeptides
  • C07K5/06008—Dipeptides with the first amino acid being neutral
  • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
  • C07K5/06034—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms
  • C07K5/06052—Val-amino acid
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/06—Dipeptides
  • C07K5/06008—Dipeptides with the first amino acid being neutral
  • C07K5/06078—Dipeptides with the first amino acid being neutral and aromatic or cycloaliphatic
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/06—Dipeptides
  • C07K5/06104—Dipeptides with the first amino acid being acidic
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/06—Dipeptides
  • C07K5/06139—Dipeptides with the first amino acid being heterocyclic

Definitions

• the present invention relates to chemical compounds and their pharmaceutical uses. More particularly, the invention relates to selective inhibitors of caspases and their uses for the prevention and/or treatment of various diseases and conditions in subjects.
• Caspases comprise a family of cysteine protease enzymes with a well-known role as key mediators in apoptosis signaling pathways and cell disassembly, lnterleukin converting enzyme (ICE), also known as Caspase-1 , was the first identified caspase. In humans, 11 other known caspases have been further identified. Caspases have been classified in two general groups according to their effects: proapoptotic (caspase-2, 3, 6, 7, 8, 9, 10) and proinflammatory (caspase-1 , 4, 5, 11, 12) caspases.
• the proapoptotic caspases have been divided in initiators (caspase-2, 8, 9, 10) also known as group II, and executioners (caspase-3, 6, 7) of the apoptotic process or group III.
• the lnterleukin converting enzyme (ICE) also known as Caspase-1 has a proinflammatory role only.
• caspase-1 plays an important role in response to pathogenic infection as well as in inflammatory and autoimmune disorders.
• caspase-1 activity is increased in retinas of diabetic patients and it constitutes a critical regulator of cardiomyocyte programmed cell death in the mammalian heart.
• Caspases also plays a role in neurodegenerative diseases and trauma. For instance, it has been shown that the caspase-3 cascade is highly activated due to the traumatic spinal cord injury.
• ALS Amyotrophic Lateral Sclerosis
• caspase-7, -8, and -9 in a mouse model at end stage of ALS
• Increased levels of apoptosis and caspase activity are reported to be frequently observed at sites of cellular damage in both acute (e.g. Sepsis, myocardial infarction(MI), Ischemic Stroke, Spinal cord injury (SCI), traumatic Brain Injury (TBI)) and neurodegenerative disease (e.g. Alzheimer's, Parkinson's and Huntington's diseases, and multiple sclerosis (MS)).
• MI myocardial infarction
• Ischemic Stroke Spinal cord injury
• TBI traumatic Brain Injury
• MS multiple sclerosis
• caspases are involved in a number of diseases, several compounds and methods have been developed to inactivate them.
• the broad irreversible caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (z-VAD-fmk) was protective and efficiently blocked death receptor-mediated liver injury in animal models (Rodriguez et al. (1996), J Exp Med. 1996 Nov 1 ; 184(5):2067-72). Myocardial infarction and the resulting death of myocytes was ameliorated by z-VAD-fmk and related peptide inhibitors in animal models (Yaoita et al., 91998) Circulation 97: 276-281).
• the present invention addresses these needs for novel therapies, new treatment methods, compounds, and pharmaceutical compositions.
• the present invention relates to compounds according to any of Formula I, IA, II, MA, III, or IHA as defined herein, compositions thereof and methods for the prevention and/or treatment of caspase-related diseases in subjects.
• Particular aspects of the invention relates to use of compounds according to any of Formula I, IA, II, MA, III, or MIA as defined herein.
• One aspect of the invention concerns a method for preventing and/or treating a caspase- related disease in a subject in need thereof, comprising administering to said subject an effective amount of a compound represented by any of Formula I, IA, II, HA, III, or IHA as defined herein.
• One aspect of the invention concerns the use of a compound a compound represented by any of Formula I, IA, II, NA, III, or MIA as defined herein for preventing and/or treating of caspase-related diseases in a subject in need thereof.
• Another related aspect of the invention concerns the use of a compound represented by any of Formula I, IA, II, MA, III, or IHA as defined herein for the manufacture of a medication for preventing and/or treating of caspase-related diseases in a subject in need thereof.
• One aspect ot he invention concerns a method of treating excessive apoptosis affected by caspase activity in a cell or a tissue, the method comprising: contacting the cell or tissue with an effective amount of one or more compounds represented by any of Formula I, IA, II, HA, III, or HIA as defined herein, so as to treat the excessive apoptosis.
• One particular aspect of the invention concerns the use of a compound represented by any of Formula I, IA, II, MA, III, or IHA as defined herein for use in apoptosis mediated diseases.
• Caspase-related disease as defined herein are selected from the group consisting of apoptosis mediated diseases, IL-1 mediated diseases, inflammatory diseases, autoimmune diseases, autoinflammatory diseases, proliferative diseases, infectious diseases, degenerative diseases, retinal disorders, inflammatory peritonitis, osteoarthritis, pancreatitis, asthma, respiratory distress syndrome, rheumatoid arthritis, systemic lupus erythematous, scleroderma, Grave's disease, autoimmune gastritis, diabetes, autoimmune hemolytic anemia, autoimmune neutropenia, thrombocytopenia, hepatitis, inflammatory bowel disease, Crohn's disease, psoriasis, dermatitis, Graft vs host disease, organ transplant rejection, osteoporosis, leukemias and related disorders, multiple myeloma-related diseases, metastatic melanomas, Kaposi's sarcoma, sepsis, septic shock, Alzheimer's disease
• the invention also provides methods and strategies of targeting caspases.
• the approach consists of designing a suicide substrate leading to a permanent inhibition of the caspase.
• the approach consists of designing a substrate that is recognizable enough for caspases, especially one or more specific caspase(s), to fit into it, to be potentially cleaved at a specific position in a way that makes the caspase enzyme irreversibly linked to the substrate thereby leading to a permanent inhibition of the caspase.
• the suicide substrates of this invention are vinyl electron withdrawing group (EWG).
• the present inventors have discovered compounds that have beneficial pharmaceutical properties and that these compounds may be effective for use in caspase-mediated diseases such as sepsis, myocardial infarction, ischemic stroke, spinal cord injury (SCI) 1 traumatic brain injury (TBI) and neurodegenerative disease (e.g. multiple sclerosis (MS) and Alzheimer's, Parkinson's, and Huntington's diseases).
• caspase-mediated diseases such as sepsis, myocardial infarction, ischemic stroke, spinal cord injury (SCI) 1 traumatic brain injury (TBI) and neurodegenerative disease (e.g. multiple sclerosis (MS) and Alzheimer's, Parkinson's, and Huntington's diseases).
• the invention concerns a compound represented by Formula I
• A, PX, P5, P4, P3, P2, R 1 , R 2 , a and b are as defined hereinabove and hereinbelow; or a prodrug, or a pharmaceutically acceptable salt to allow the drug to penetrate the cell membrane or the compound is labeled with a detectable label or an affinity tag thereof.
• the line "-" when located between P2, P3, P4, P5 and PX represents a peptide bond or a peptidomimetic bond;
• the PX, P5, P4, P3, P2 amino acid residues are normally linked via a peptide bond, that is, a peptidic carbamoyl group, i.e. -CONH--.
• peptidomimetic bonds are also contemplated, such as CH 2 -NH, CO-CH 2 , azapeptide and retro-inverso bonds.
• R 1 and R 2 that are bonded to the vinyl group can be either in the cis configuration or the trans configuration, as represented by the wavy lines.
• R 1 is configured to be trans such that the electron withdrawing capability of the R 1 group is stabilized.
• A, AA x , AA 5 , AA 4 , AA 3 , AA 2 R 1 , R 2 , a and b are as defined hereinabove and hereinbelow; or a prodrug, or a pharmaceutically acceptable salt to allow the drug to penetrate the cell membrane, or the compound is labeled with a detectable label or an affinity tag thereof.
• the present invention includes compounds of Formula Il :
• NA wherein A, AA 4 , AA 3 , AA 2 R 1 , and R 2 are as defined hereinabove and hereinbelow.
• One subset of compounds of Formula III includes compounds of Formula IMA :
• MIA MIA wherein A, AA 5 , AA 4 , AA 3 , AA 2, R 1 , and R 2 are as defined hereinabove and hereinbelow.
• a and b are both 0. In another example, a is 0 and b is 1.
• A is H.
• A is R 3 -OC(O)-.
• A is PhCH 2 OC(O)-.
• R 1 is an electron withdrawing group (EWG) selected from
• aryl, the heteroaryl, or the heterocyclyl are optionally substituted with one or more R 30 .
• R 2 is
• R 1 , R 6 , R 7 , R 8 and R 9 are as defined hereinabove and hereinbelow.
• R 2 is H.
• R 2 is halogen
• R 2 is Cl.
• R 4 is
• R 4 is
• R 4 is H.
• R 5 is
• heterocyclyl or 8) any optionally protected (D) or (L) amino acid residue.
• R 6 is
• alkyl or the cycloalkyl are optionally substituted with one or more R 10 substituents; and in which the aryl, heteroaryl or heterocyclyl are optionally substituted with one or more R 20 substituents.
• R 7 and R 8 are independently selected from: 1) H,
• alkyl and the cycloalkyl are optionally substituted with one or more R 10 substituents, and the aryl, the heteroaryl and the heterocyclyl are optionally substituted with one or more R 20 substituents.
• R 9 is
• heterocyclyl in which the alkyl or the cycloalkyl are optionally substituted with one or more R 10 substituents; and in which the aryl, heteroaryl or heterocyclyl are optionally substituted with one or more R 20 substituents.
• R 10 is independently selected from:
• R 7 , R 8 , R 9 are as defined hereinabove and hereinbelow; m is an integer of O, 1 , or 2.
• R 20 is independently selected from:
• alkyl and the cycloalkyl are optionally substituted with one or more R 6 substituents; and wherein the aryl, the heteroaryl, or the heterocyclyl are optionally substituted with one or more R 30 .
• R 7 , R 8 and m are as defined hereinabove and hereinbelow.
• R 30 is
• R 5 and R 20 are as defined hereinabove and hereinbelow.
• the present invention includes compounds of Formula MA:
• AA 2 is the amino acid side chain of VaI, Leu, Pro, Met, Ala, Thr, His.
• AA 3 is the amino acid side chain of Trp, Tyr, Ala, Asp, GIu, GIn, Phe, Ser, Thr, VaI, Tyr, GIy, Leu; or AA 3 is phenylglycine, indanylglycine, or Ala-(2'-quinolyl);
• AA 4 is the amino acid side chain of Asp
• R 1 , R 2 and R 4 are as defined hereinabove and hereinbelow.
• the present invention includes compounds of Formula MA:
• AA 2 is the amino acid side chain of Thr, His, VaI, Trp, He, or Ala
• AA 3 is the amino acid side chain of GIu or AA 3 is Ala-(2'-quinolyl);
• AA 4 is the amino acid side chain of lie, Leu, GIu, Asp, Ala, Pro or VaI;
• R 1 , R 2 and R 4 are as defined hereinabove and hereinbelow.
• the present invention includes compounds of Formula MIA
• AA 2 is the amino acid side chain of Ala, Ser, Lys or VaI;
• AA 3 is the amino acid side chain of VaI, GIu, Thr, or GIn;
• AA 4 is the amino acid side chain of Asp, or Leu;
• AA 5 is the amino acid side chain of VaI or Leu;
• R 1 , R 2 and R 4 are as defined hereinabove and hereinbelow.
• the present invention includes compounds of Formula MA (caspase 1 inhibitors)
• AA 2 is the amino acid side chain of VaI, Ala, Thr, or His;
• AA 3 is the amino acid side chain of GIu, GIn, Asp, Ala, GIy, Thr, VaI, Trp; or AA 3 is phenylglycine or indanylglycine;
• AA 4 is the amino acid side chain of Tyr, Trp, Phe, or Asp;
• R 1 , R 2 and R 4 are as defined hereinabove and hereinbelow.
• the term “comprising” is intended to mean that the list of elements following the word “comprising” are required or mandatory but that other elements are optional and may or may not be present 0
• the term “consisting of” is intended to mean including and limited to whatever follows the phrase “consisting of Thus the phrase “consisting of indicates that the listed elements are required or mandatory and that no other elements may be present
• alkyl is intended to include both branched and straight chain 5 saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, for example, C 1 -C 6 as in C 1 -C 6 - alkyl is defined as including groups having 1 ,2,3,4,5 or 6 carbons in a linear or branched arrangement
• Examples of Ci-C ⁇ -alkyl and C 1 -C 4 alkyl as defined above include, but are not limited to, methyl, ethyl, n-propyl, /-propyl, n-butyl, f-butyl, /-butyl, pentyl and hexyl 0
• alkenyl is intended to mean unsaturated straight or branched chain hydrocarbon groups having the specified number of carbon atoms therein, and in which at least two of the carbon atoms are bonded to each other by a double bond, and having either E or Z regiochemistry and combinations thereof
• cycloalkyl is intended to mean a monocyclic saturated aliphatic hydrocarbon group having the specified number of carbon atoms therein, for example, C 3 -C 7 as in C 3 -C 7 cycloalkyl is defined as including groups having 3,4,5,6, or 7 carbons in a monocyclic arrangement.
• Examples of C 3 -C 7 cycloalkyl as defined above include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.
• halo or halogen is intended to mean fluorine, chlorine, bromine and iodine.
• haloalkyl is intended to mean an alkyl as defined above, in which each hydrogen atom may be successively replaced by a halogen atom.
• haloalkyls include, but are not limited to, CH 2 F, CHF 2 and CF 3 .
• aryl either alone or in combination with another radical, means a carbocyclic aromatic monocyclic group containing 6 carbon atoms which may be further fused to a second 5- or 6-membered carbocyclic group which may be aromatic, saturated or unsaturated.
• Aryl includes, but is not limited to, phenyl, indanyl, 1- naphthyl, 2-naphthyl and tetrahydronaphthyl.
• the aryls may be connected to another group either at a suitable position on the cycloalkyl ring or the aromatic ring.
• heteroaryl is intended to mean a monocyclic or bicyclic ring system of up to ten atoms, wherein at least one ring is aromatic, and contains from 1 to 4 hetero atoms selected from the group consisting of O, N, and S.
• the heteroaryl substituent may be attached either via a ring carbon atom or one of the heteroatoms.
• heteroaryl groups include, but are not limited to thienyl, benzimidazolyl, benzo[b]thienyl, furyl, benzofuranyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl, 2H- pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolyl, phthalazinyl, napthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl, isothiazolyl, isochromanyl, chromanyl,
• heterocycle As used herein, the term “heterocycle”, “heterocyclic” or “heterocyclyl” is intended to mean a 5, 6, or 7 membered non-aromatic ring system containing from 1 to 4 heteroatoms selected from the group consisting of O, N and S.
• heterocycles include, but are not limited to pyrrolidinyl, tetrahydrofuranyl, piperidyl, pyrrolinyl, piperazinyl, imidazolidinyl, morpholinyl, imidazolinyl, pyrazolidinyl, and pyrazolinyl.
• EWG electron withdrawing group
• the EWG is conjugated with the alkene bond, such that the electron withdrawing properties of the EWG allow nucleophilic attack by a caspase at the alkene bond, i.e. the alkene bond and the EWG are electronically conjugated.
• the covalent bond between the alkene bond and the EWG is a direct one, without intervening moieties that would prevent the electron withdrawing properties of the EWG from being exerted on the alkene bond.
• the term "detectable label” is intended to mean a group that may be linked to a compound of the present invention to produce a probe or to a caspase, such that when the probe is associated with the caspase, the label allows either direct or indirect recognition of the probe so that it may be detected, measured and quantified.
• affinity tag is intended to mean a ligand or group, which is linked to either a compound of the present invention or to a caspase to allow another compound to be extracted from a solution to which the ligand or group is attached.
• the term "probe” is intended to mean a compound of Formula I, IA, II, MA, III, or IMA, which is labeled with either a detectable label or an affinity tag, and which is capable of binding, either covalently or non-covalently, to a caspase.
• the probe When, for example, the probe is non-covalently bound, it may be displaced by a test compound.
• the probe When, for example, the probe is bound covalently, it may be used to form cross-linked adducts, which may be quantified and inhibited by a test compound.
• the term "optionally substituted with one or more substituents” or its equivalent term “optionally substituted with at least one substituent” is intended to mean that the subsequently described event of circumstances may or may not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not.
• the definition is intended to mean from zero to five substituents.
• the substituents themselves are incompatible with the synthetic methods of the present invention, the substituent may be protected with a suitable protecting group (PG) that is stable to the reaction conditions used in these methods. The protecting group may be removed at a suitable point in the reaction sequence of the method to provide a desired intermediate or target compound.
• PG protecting group
• Suitable protecting groups and the methods for protecting and de-protecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples of which may be found in T. Greene and P. Wuts, Protecting Groups in Chemical Synthesis (3 rd ed.), John Wiley & Sons, NY (1999), which is incorporated herein by reference in its entirety.
• Examples of protecting groups used throughout include, but are not limited to Fmoc, Bn, Boc, CBz and COCF 3 .
• a substituent may be specifically selected to be reactive under the reaction conditions used in the methods of this invention. Under these circumstances, the reaction conditions convert the selected substituent into another substituent that is either useful in an intermediate compound in the methods of this invention or is a desired substituent in a target compound.
• a series of non natural amino acids which may be used in place of natural amino acids is but not limited to 3-am ⁇ no 2-hydroxy pyridine, (2furyl)alan ⁇ ne, 1 amino Icyclohexane carboxylic acid, (2th ⁇ enyl)alan ⁇ ne, 2-Am ⁇ nobenzo ⁇ c acid (2-Abz), 2Py ⁇ dylAlan ⁇ ne, IAmino I Cyclopentanecarboxilic acid, 2-Am ⁇ nobutyr ⁇ c acid (2Abu), 3Am ⁇ no3phenylprop ⁇ on ⁇ c acid, Aminocyclopentane carboxylic acid (ACPC), 4-Am ⁇ nomethylbenzo ⁇ c acid (Amb), Aminoisobutiric acid (Aib), p-Benzoyl-1 -phenylalanine (Bpa), AllyIGlycine, 4-Am ⁇ nomethyl cyclohexane carboxylic acid (Amc), Cyclohexyl-alanine (Cha), delta
• the term "residue" when referring to ⁇ -am ⁇ no acids is intended to mean a radical derived from the corresponding ⁇ -am ⁇ no acid by eliminating the hydroxyl of the carboxy group and one hydrogen of the ⁇ -am ⁇ no group
• the terms GIn, Ala, GIy, lie, Arg, Asp, Phe, Ser, Leu, Cys, Asn, and Tyr represent the residues of L- glutamine, L-alanine, glycine, L-isoleucine, L-arginine, L-aspartic acid, L-phenylalanine, L-se ⁇ ne, L-leucine, L-cysteine, L-asparagine, and L-tyrosine, respectively
• amino acid side chain is intended to mean the part of an amino acid's chemistry that differentiates it from other amino acids
• Amino acid structure includes a carboxyl group, an amine group plus the individual side chain
• Each amino acid has a unique side chain This is applied to unatural amino acids as well This side chain may exist in protected form or not
• prodrug is intended to mean a compound that may be converted under physiological conditions or by solvolysis to a biologically active compound of the present invention
• prodrug refers to a precursor of a compound of the invention that is pharmaceutically acceptable.
• a prodrug may be inactive or display limited activity when administered to a subject in need thereof, but is converted in vivo to an active compound of the present invention.
• prodrugs are transformed in vivo to yield the compound of the invention, for example, by hydrolysis in blood or other organs by enzymatic processing.
• the prodrug compound often offers advantages of solubility, tissue compatibility or delayed release in the subject (see, Bundgard, H., Design of Prodrugs (1985), pp.
• prodrug includes any covalently bonded carriers which release the active compound of the invention in vivo when such prodrug is administered to a subject.
• Prodrugs of a compound of the present invention may be prepared by modifying functional groups present in the compound of the invention in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to a parent compound of the invention.
• pharmaceutically acceptable salt is intended to mean both acid and base addition salts.
• the term "pharmaceutically acceptable acid addition salt” is intended to mean those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.
• inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like
• organic acids such as acetic acid,
• salts derived from inorganic bases include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like.
• Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like.
• basic ion exchange resins such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine,
• the compounds of the present invention, or their pharmaceutically acceptable salts may contain one or more asymmetric centers, chiral axes and chiral planes and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms and may be defined in terms of absolute stereochemistry, such as (R)- or (S)- or, as (D)- or (L)- for amino acids.
• the present invention is intended to include all such possible isomers, as well as, their racemic and optically pure forms.
• Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as reverse phase HPLC.
• the racemic mixtures may be prepared and thereafter separated into individual optical isomers or these optical isomers may be prepared by chiral synthesis.
• the enantiomers may be resolved by methods known to those skilled in the art, for example by formation of diastereoisomeric salts which may then be separated by crystallization, gas-liquid or liquid chromatography, selective reaction of one enantiomer with an enantiomer specific reagent. It will also be appreciated by those skilled in the art that where the desired enantiomer is converted into another chemical entity by a separation technique, an additional step is then required to form the desired enantiomeric form. Alternatively specific enantiomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts, or solvents or by converting one enantiomer to another by asymmetric transformation.
• Certain compounds of the present invention may exist as a mix of epimers.
• Epimers means diastereoisomers that have the opposite configuration at only one of two or more stereogenic centres present in the respective compound.
• Certain compounds of the present invention may exist in Zwitterionic form and the present invention includes Zwitterionic forms of these compounds and mixtures thereof.
• the compounds of the invention also may exist in hydrated and anhydrous forms. Hydrates of the compound of any of the formulas described herein are included as compounds of the invention.
• the compound according to any of the formulas described herein is a monohydrate.
• the compound of the invention comprises about 10% or less, about 9 % or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2% or less, about 1% or less, about 0.5% or less, about 0.1 % or less by weight of water.
• the compounds of the invention comprise, about 0.1 % or more, about 0.5% or more, about 1% or more, about 2% or more, about 3% or more, about 4% or more, about 5% or more, or about 6% or more by weight of water.
• the starting materials and reagents used in preparing these compounds are either available from commercial suppliers such as Sigma-Aldrich Chemicals, Anaspec or chemipex.
• the starting materials and the intermediates of the reaction may be isolated and purified if desired using conventional techniques such chromatography (Biotage flash chromatography), filtration, distillation, etc. Such materials can be characterized using conventional analytical methods such as NMR and LCMS.
• the coupling step is carried out in the presence of a suitable coupling agent such as, but not limited to, diisopropyl carbodiimide (DPC), anhydride mixte (isobutylchloroformate),0-benzotriazole-1-yloxytrispyrrolidinophosphonium hexafluorophosphate(PyBOP), O-benzotriazol-1-yl- ⁇ /, ⁇ /, ⁇ /, ⁇ / tetramethyluroniumhexafluoro-phosphate (HBTU), O-(7-azabenzotriazol-1 -yl)-1 , 1 ,3,3- tetramethyluronium hexafluoro-phosphate (HATU), 1-hydroxybenzotriazole (HOBT) in the presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC).
• DPC diisopropyl carbodiimide
• a base such as N,N-diisopropylethylamine, triethyl amine, or N-methylmorpholine.
• the reaction is carried out at 20 0 C except for anhydride mix formation (isobutyl chloroformate, -5/-13 0 C) to avoid any racemisation of amino acids.
• Removal of the amino protecting group is carried out but by piperidine in dichloromethane (Fmoc protecting group).
• the removal of ⁇ -tert-butyl carboxylic acid is carried out by TFA.
• Selective removal of N-Boc from B-tert butyl aspartic acid is carried out by PTSA or TFA at O 0 C.
• Vinyl suifone function is elucidated as an example only in the following examples, but is not limited to it. It will be recognized by a skilled person in the art, that other vinyl electron-withdrawing group (EWG) may be used in this invention.
• EWG vinyl electron-withdrawing group
• Benzyloxycarbonyl (Z) is elucidated as an example of linkers that may facilitate the penetration of the drug into cells. It is recognized that other X-R3 may be used as well.
• the allyl group was introduced directly by the use of commercially available AA 2 -OAIIyI protected form or synthesized from the the corresponding amino
• a compound of Formula 1a can be prepared by the procedure described bellow
• allyl group was introduced directly by the use of commercially available AA 2 -OAIIyI protected form or synthesized from the protected amino acid fmoc-AA 2 -OH and allyl alcohol (EDC, DMAPcat, NMM, CH 2 CI 2 /DMF (5/1)) followed with fmoc deprotection (piperidine, CH 2 CI 2 ).
• the AA 2 -OAIIyI is coupled to Fmoc-AA 3 -OH with the coupling reagent 1-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and subsequent fmoc deprotection with piperidine gives AA 3 -AA 2 -OaIIyI (intermediate A) ready for coupling with (Z) AA 4 -OH gave (Z) AA 4 -AA 3 -AA 2 -OaIIyI (intermediate B) which upon removal of allyl group (tetrakis) liberate the C-terminal carboxylic acid (tripeptide A) as described in the following schem. CH 2 CI 2
• the suicide substrate proposed in the following scheme is Asp ⁇ -chlorovinyl methylsulfone.
• Boc-Asp(B-tert-butyl)-H is synthesized from Boc-Asp(B-tert- butyl)-N-hydroxysuccinimide ester as reported by (Mancuso A et al., 1981 , William R. Ewing et al., 1999 and Won Bum Jang. 2004).
• a compound of Formula 1 b can be prepared by the procedure described bellow
• the suicide substrate proposed in the following scheme is Asp ⁇ -chlorovinyl phenylsulfone.
• a compound of Formula 1c can be prepared by the procedure described bellow
• the suicide substrate proposed in the following schem is Asp ⁇ -methoxyvinyl methylsulfone.
• Oxydation of diethyl methylthiomethylphosphonate include anodic oxidation in NaOMe/MeOH (saturated with CO2) to give the O, S-acetal of diethoxyphosphinylformaldehyde, which upon oxidation may give the corresponding sulfone as Diethyl methoxy(methylsulfone)phenyl phosphonate (Shankaran, K et al. 2001).
• a compound of Formula 1d can be prepared by the procedure described bellow
• the suicide substrate proposed in the following schem is Asp(B-Methyl) methyl vinylsulfone.
• Boc-Asp(B-Methyl)-N-hydroxysuccinimide ester was reduced to the corresponding alcohol (NaBH 4 , THF), a subsequent oxidation (oxalyl chloride , DMSO) gave the aldehyde: Boc-Asp(B-Methyl)-H.
• Boc-Asp(B-Methyl)-H Treatment of Boc-Asp(B-Methyl)-H with sodium anion of Diethyl (methylsulfone) methyl phosphonate results in the corresponding Boc-Asp ( ⁇ -Methyl) methyl vinylsulfone in the manner of Wadsworth and Emmons.
• a compound of Formula 1e can be prepared by the procedure described bellow
• the suicide substrate proposed in the following schem is Asp methyl vinyllsulfone.
• Boc-Asp(B-tert-butyl)-H is synthesized from Boc-Asp(B-tert- butyl)-N-hydroxysuccinimide ester as reported by (Mancuso A et al., 1981 , William R. Ewing et al., 1999 and Won Bum Jang. 2004).
• agent A compound of Formula 1f can be prepared by the procedure described bellow
• the suicide substrate proposed in the following schem is Asp phenyl vinyllsulfone.
• Diethyl phenylsulfonylmethylphosphonate was obtained in one step from benzenesulfonyl fluoride and triethyl phosphorane in the presence of lithium hexamethyldisilazide (Won Bum Jang et al., 1998).
• a compound of Formula 1g can be prepared by the procedure described bellow
• the suicide substrate proposed in the following schem is Asp phenoxy vinyllsulfone.
• Diethyl (phenoxysulfone) methylphosphonate was obtained from methanesulfonyl phenoxy and diethyl chlorophosphonate in the presence of potassium bis(trimethylsilyl)amide. A subsequent oxidation (AcOH, H 2 O 2 ) gave the corresponding sulfone.
• a compound of Formula 1 h can be prepared by the procedure described bellow
• the suicide substrate proposed in the following schem is Asp morpholine vinyllsulfone.
• Diethyl (morpholinesulfone) methylphosphonate was prepared from methane sulfonyl morpholine and chloromethylphosphonate in the presence of potassium bis(thmethylsilyl) amide.
• a compound of Formula 1i can be prepared by the procedure bellow
• the suicide substrate proposed in the following schem is Asp(B-Methyl)- vinylsulfone derivatives.
• Boc-Asp(B-Methyl)-N-hydroxysuccinimide ester was reduced to the corresponding alcohol (NaBH 4 , THF), a subsequent oxidation (oxalyl chloride , DMSO) gave the aldehyde Boc-Asp(B-Methyl)-H.
• Boc-Asp(B-Methyl)-H Treatment of Boc-Asp(B-Methyl)-H with sodium anion of Diethyl (methylsulfone) phosphonate derivatives results in the corresponding Boc-Asp ( ⁇ -Methyl) vinylsulfone derivatives in the manner of Wadsworth and Emmons.
• the compounds of the present invention are represented by generalized Formula I, IA, II, HA, III, or IHA, or a pharmaceutically acceptable salt and/or prodrug thereof:
• An additional aspect of the invention relates to a method for designing caspase inhibitors. Following the same approach as outlined hereinafter, those skilled in the art will appreciate that it is conceivable to further improve the potency of the compounds of Formula II, e.g. Z-Asp-Phg-Val-AspVSmethyl (53), against caspase-3 and inhibit selectively additional caspases such as caspase-2, caspase-8, caspase-9 and caspase- 1 .
• caspase-3 requires an additional Asp at position P4, which confers caspase-3 its specificity:
• suicide substrates such as AspVSmethyl (Compound 93) and Asp(Ofbu)VSmethyl (Compound 33) are devoid of any activity against caspase-
• the substrate z-Asp(Ofbu)-Phg-val-OH (Compound 16) is also devoid of any activity against caspase-3. However the fusion product of peptide and suicide substrate, z-Asp-
• Phg-Val-AspVSmethyl (Compound 53), proved to be a very potent inhibitor of caspase-3 with an IC50 30-9OnM. Inhibition is selective since even though caspase-7 belongs to the same group than caspase-3, the IC50 value for this caspase was about 12 fold higher.
• the amino acids at both position P3 and P2 can serve to selectively target caspase-3 as observed but also to selectively target other caspases.
• This substitution enhanced the inhibitory effect against the group III of caspases (caspase-3 (30-9OnM) and 7 (0.18-0.30 uM) and, with about 29 fold less efficiency, against group 1 (caspase-1 (0.6-1.2 uM)).
• Trp at P3 position (z-Asp-Trp-Val- AspVSmethyl; Compound 88) retained the selectivity against caspase-3 (30-90 nM) and produced an additional activity against caspase-1 (0.6-1.2 uM). This molecule possesses therefore the capability to inhibit selectively two caspases belonging to two different groups, namely the proinflamatory and the proapoptotic group.
• the presence of Glutamic acid at P-3 position (z-Asp-Glu-Val-AspVSmethyl; Compound 59) retained the selectivity against caspase-3 (IC50 2OnM) and produced an additional activity against caspase-7 (IC50 42 nM) and caspase-9 (509 nM). This molecule therefore possesses the capability to inhibit selectively two caspases belonging to two different groups, namely initiator and executioner caspases.
• Changes in position P4 can also affect the selectivity of a given inhibitor.
• the amino acid that has been showed to fit well into the corresponding caspase-1 pockets at position P4 is Tyrosine. Therefore, z-Tyr-Val-Ala-AspVS phenyl (Compound 96) was tested against different caspases, and it proved to be selective against caspase-1 (IC50 1.2-1.5 ⁇ M).
• the amino acid that has been showed to fit well into the corresponding caspase-1 pockets at position P3 is glutamic acid. Therefore, z-Tyr-Glu-Ala-AspVS methyl (Compound 82) was tested against different caspases and the inhibition of caspase-1 was enhanced to 0.5 ⁇ M.
• the method comprises synthesizing compounds having the following general Formula D1 :
• P2 is selected from the following amino acids: V, L, P, M, A, T, and H;
• P3 is selected from the following amino acids: Phg, E, Indanylglycine, W, Y, A, D, AIa- (2'-quinolyl), Q, F, S, T, V, Y, G, L;
• D a is (D) or (L) aspartic acid.
• D b is the side chain of (D) or (L) aspartic acid.
• the following compound (DEVD-vinyl phenyl sulfone) is an example a compounds having a sequence (i.e. D a is Cbz-aspartic acid; P3 is GIu; P2 is VaI; D b is Asp; and the suicide substrate is vinyl phenyl sulfone) designed to selectively inhibit caspase-3 :
• the method comprises synthesizing compounds having the following general Formula D2:
• P2 is selected from the following amino acids: T, H, V, W, I, and A;
• P3 is selected from the following amino acids: E, Ala(2'-quinolyl;
• P4 is selected from the following amino acids: I, L, E, D, A, P, and V;
• D is the side chain of (D) or (L) aspartic acid and the suicide substrate is is vinyl phenyl sulfone.
• P4 is Cbz-L
• P3 is GIu
• P2 is His
• D is Asp
• the suicide substrate is vinyl phenyl sulfone
• the method comprises synthesizing compounds having the following general Formula D3:
• P2 is selected from the following amino acids: A, S, K and V;
• P3 is selected from the following amino acids: V, E, T and Q;
• P4 is selected from the following amino acids: D and L;
• P5 is selected from the following amino acids: V and L;
• D is the side chain of (D) or (L) aspartic acid; and the suicide substrate is selected from the group consisting of vinyl phenyl sulfone.
• VDEHD- vinylphenyl sulfone having a sequence (i.e. P5 is Cbz VaI; P4 is Asp; P3 is GIu; P2 is His and D is Asp.
• the suicide substrate is vinyl phenyl sulfone)., which is designed to selectively inhibit caspase-2:
• the method comprises synthesizing compounds having the following general Formula D4:
• P2 is selected from the following amino acids: V, A, T, and H.
• P3 is selected from the following amino acids: E, Q, D, A, G, T, V, Ala(2'-quinolyl), indanylglycine, and W;
• P4 is selected from the following amino acids: Y, W, F, and D;
• D is the side chain of (D) or (L) aspartic acid; and the suicide substrate is selected from the group consisting of vinyl phenyl sulfone.
• the compounds of the invention have beneficial pharmaceutical properties and these compounds may have pharmaceutical applications in the prevention and/or treatment of various diseases and conditions in a subject.
• Medical and pharmaceutical applications contemplated by the inventors include, but are not limited to, caspase-mediated diseases.
• the compounds of the present invention may have useful benefits on cells in vitro such as promoting cell survival or the health of the cells.
• subject includes living organisms in which blood disorders, renal failure, inflammatory-related diseases associated with high blood pressure, and/or oxidative stress-related disorders, can occur, or which are susceptible to such conditions.
• subject includes animals (e.g., mammals (e.g., cats, dogs, horses, pigs, cows, goats, sheep, rodents (e.g., mice or rats), rabbits, squirrels, bears, primates (e.g., chimpanzees, monkeys, gorillas, and humans)), as well as avians (e.g. chickens, ducks, Peking ducks, geese), and transgenic species thereof.
• the subject is a mammal. More preferably, the subject is a human. Even more preferably, the subject is a human patient in need of treatment.
• caspase-mediated disease includes all diseases, disorder and/or conditions in which any one or more of caspase-1 , -2, -3, -4, -5, -6, -7, -8, -9, -10, -11 , -12, plays a significant role.
• the caspase-mediated disease mainly involves executioner caspases (caspase-3, 6, 7).
• the caspase-mediated disease mainly involves initiators (caspase-2, 8, 9, 10).
• a compound of the invention shows a high specificity towards one particular caspase.
• a compound of the invention is able to inhibit two groups of caspases.
• a compound of the invention even is able to inhibit two specific caspases belonging to two different groups of caspases.
• caspase-mediated disease includes, but are not limited to, apoptosis mediated diseases, IL-1 mediated diseases, inflammatory diseases, autoimmune diseases, autoinflammatory diseases, proliferative diseases, infectious diseases, degenerative diseases, retinal disorders, inflammatory peritonitis, osteoarthritis, pancreatitis, asthma, respiratory distress syndrome, rheumatoid arthritis, systemic lupus erythematous, scleroderma, Grave's disease, autoimmune gastritis, diabetes, autoimmune hemolytic anemia, autoimmune neutropenia, thrombocytopenia, hepatitis, inflammatory bowel disease, Crohn's disease, psoriasis, dermatitis, Graft vs host disease, organ transplant rejection, osteoporosis, leukemias and related disorders, multiple myeloma-related diseases, metastatic melanomas, Kaposi's sarcoma, sepsis, septic shock, Alzheimer's disease
• preventing or “prevention” is intended to refer to at least the reduction of likelihood of the risk of (or susceptibility to) acquiring a disease or disorder (i.e., causing at least one of the clinical symptoms of the disease not to develop in a patient that may be exposed to or predisposed to the disease but does not yet experience or display symptoms of the disease).
• Biological and physiological parameters for identifying such patients are provided herein and are also well known by physicians.
• treatment includes the application or administration of a compound of the invention to a subject (or application or administration of a compound of the invention to a cell or tissue from a subject) with the purpose of stabilizing, curing, healing, alleviating, relieving, altering, remedying, less worsening, ameliorating, improving, or affecting the disease or condition, the symptom of the disease or condition, or the risk of (or susceptibility to) the disease or condition.
• treating refers to any indicia of success in the treatment or amelioration of an injury, pathology or condition, including any objective or subjective parameter such as abatement; remission; lessening of the rate of worsening; lessening severity of the disease; stabilization, diminishing of symptoms or making the injury, pathology or condition more tolerable to the subject; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; or improving a subject's physical or mental well-being.
• the term “treating” can include increasing a subject's life expectancy and/or delay before additional treatments are required.
• the present invention relates to methods, compounds and compositions for prevention and/or treatment of a caspase-mediated disease in a subject, preferably a human patient in need thereof.
• Another aspect of the invention relates to the use of the compounds described herein for inhibiting a caspase or a caspase-like protein in a cell, comprising contacting the caspase or caspase-like protein with an effective amount of a caspase inhibitor according to the invention.
• the subject may be suffering from a viral infection. Therefore, the invention also relates to a method for the prophylaxis or therapy of a viral infection, comprising administering to a subject in need thereof an effective dose of a caspase inhibitor according to the invention (or a pharmaceutical composition comprising the same). This may be helpful for inhibiting a cellular caspase thereby inhibiting virus multiplication.
• Also of particular interest is a method for the treatment of excessive apoptosis affected by caspase activity in a cell or a tissue, comprising contacting the cell or the tissue with an effective amount of one or more caspase inhibitor according to the invention (or a pharmaceutical composition comprising the same).
• the method for culturing a large quantity of stem cells may involves an effective amount of one or more caspase inhibitor according to the invention (or a pharmaceutical composition comprising the same) and a medium for culturing stem cells.
• the compounds of the invention be also effective in inhibiting additional families of proteases, including but not limited to, serine peptidases, cysteine peptidases, aspartic peptidases, metallo-peptidases, and other peptidases of unknown catalytic type.
• proteases including but not limited to, serine peptidases, cysteine peptidases, aspartic peptidases, metallo-peptidases, and other peptidases of unknown catalytic type.
• ZBIGNIEW GRZONKA Cysteine protease. Industrial Enzymes, 181- 195, Chapter 11 , 2007 Springer.
• serial measurements can be determined. Quantitative assessment of caspase functions and parameters of caspase dysfunction are well known in the art. Examples of assays for the determination of caspases activity are provided in the Exemplification section. The compounds according to the invention can be further analyzed, tested or validated for their ability to cross the Blood Brain Barrier BBB is so desired. Many in-vitro, in-vivo and in-silico methods may be employed during drug development to mimic the BBB (Lohmann et al. (2002) Predicting blood-brain barrier permeability of drugs: evaluation of different in vitro assays.
• In-vitro models include primary endothelial cell culture and immortalized cell lines such as Caco-2, BMEC, MDCK. These cells are useful as a screening method and can appropriately rank compounds in order of BBB permeability.
• In vivo models such as the internal carotid artery single injection or perfusion, intravenous bolus injection, brain efflux index and intracerebral microdialysis provide more accurate information regarding brain uptake, and these can be complemented with novel imaging techniques (such as magnetic resonance imaging and positron emission tomography), although such methods are not suited to high-throughput permeability assessment.
• novel imaging techniques such as magnetic resonance imaging and positron emission tomography
• At least some of the prodrugs administered generates the corresponding pharmaceutical compound only after absorption by the gastrointestinal tract and/or only once it has reached the brain, i.e. after it has passed the blood brain barrier (BBB).
• BBB blood brain barrier
• a related aspect of the invention concerns pharmaceutical compositions comprising one or more of the compounds of the invention described herein.
• the compounds of the invention may be useful in preventing and/or treating caspase- mediated disease, and more particularly sepsis, myocardial infarction, ischemic stroke, spinal cord injury (SCI), traumatic brain injury (TBI) and neurodegenerative disease (e.g. multiple sclerosis (MS) and Alzheimer's, Parkinson's, and Huntington's diseases.
• the term "therapeutically effective amount' means the amount of compound that, when administered to a subject for treating or preventing a particular disorder, disease or condition, is sufficient to effect such treatment or prevention of that disorder, disease or condition. Dosages and therapeutically effective amounts may vary for example, depending upon a variety of factors including the activity of the specific agent employed, the age, body weight, general health, gender, and diet of the subject, the time of administration, the route of administration, the rate of excretion, and any drug combination, if applicable, the effect which the practitioner desires the compound to have upon the subject and the properties of the compounds (e.g. bioavailability, stability, potency, toxicity, etc), and the particular disorder(s) the subject is suffering from.
• the properties of the compounds e.g. bioavailability, stability, potency, toxicity, etc
• the therapeutically effective amount may depend on the subject's blood parameters (e.g. lipid profile, insulin levels, glycemia), the severity of the disease state, organ function, or underlying disease or complications.
• blood parameters e.g. lipid profile, insulin levels, glycemia
• Such appropriate doses may be determined using any available assays including the assays described herein.
• a physician may for example, prescribe a relatively low dose at first, subsequently increasing the dose until an appropriate response is obtained.
• pharmaceutical composition refers to the presence of at least one compound of the invention according to any one of Formula I, IA, II, NA, III, or MIA as defined herein and at least one pharmaceutically acceptable vehicle.
• pharmaceutically acceptable vehicle examples include the compounds in Table 1 and pharmaceutically acceptable salts thereof.
• “Pharmaceutically acceptable vehicle” refers to a diluent, adjuvant, excipient, or carrier with which a compound is administered.
• pharmaceutically acceptable refers to drugs, medicaments, inert ingredients etc., which are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, incompatibility, instability, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio. It preferably refers to a compound or composition that is approved or approvable by a regulatory agency of the Federal or state government or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals and more particularly in humans.
• the pharmaceutically acceptable vehicle can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils.
• a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils.
• compositions include, but are not limited to: Water for Injection USP; aqueous vehicles such as, but not limited to, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.
• aqueous vehicles such as, but not limited to, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection
• water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glyco
• Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like.
• isotonic agents are included, for example, sugars, sodium chloride, or polyalcohols such as mannitol and sorbitol, in the composition.
• Prolonged absorption of injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate or gelatin.
• Nanoparticles, liposomes, and antibodies conjugated to nanoparticles or combinations thereof, are also contemplated as pharmaceutically acceptable vehicles.
• compositions of the invention comprise an effective amount of a compound of the Formula I, IA, II, MA, III, or IMA as described hereinbefore, preferably compound Z-Asp-lndanylglycine-Val- Aspmethyl vinyl sulfone (57); Z-Asp- Glu-Val-Aspmethyl vinyl sulfone (59) Z-Asp-Ala(2'-quinolyl)-Val- Aspmethyl vinyl sulfone (55); Z-Asp-Phg-Val- Aspmethyl vinyl sulfone (53); Z-Asp-Ala(2'-quinolyl)-Val-Asp- ⁇ chlorovinyl methylsulfone (48); Z-Asp( ⁇ -methyl)-lndanylglycine-Val-Asp( ⁇ - methyl)methyl vinyl sulfone (51) Z-Asp-Tyr-Val-Aspmethyl vinyl sulfone (76)
• the invention pertains to pharmaceutical compositions for preventing and/or treating diseases or other medical conditions in which at least one caspase is significantly involved that include one or more compounds of Formula I, IA, II, HA, III, or MIA as defined herein.
• the invention pertains to pharmaceutical compositions for preventing and/or treating diseases or other medical conditions in which at least one caspase is significantly involved, the composition comprising one or more compounds of Formula I, IA, II, HA, III, or MIA as defined herein.
• the compounds of the invention may be formulated prior to administration into pharmaceutical compositions using available techniques and procedures.
• the pharmaceutical compositions may be formulated into suitable administration (orally, parenterally, (intravascular (IV), intraarterial (IA), intramuscular (IM), depo-IM, subcutaneous (SC), and depo SC), sublingually, intranasally (inhalation), intrathecally, topically, or rectally.
• the compound(s) of the invention can be orally administered.
• the formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. Methods of preparing these formulations or compositions include the step of bringing into association a compound of the present invention with a pharmaceutically acceptable vehicle (e.g. an inert diluent or an assimilable edible carrier) and, optionally, one or more accessory ingredients.
• a pharmaceutically acceptable vehicle e.g. an inert diluent or an assimilable edible carrier
• the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.
• the amount of the therapeutic agent in such therapeutically useful compositions is such that a suitable dosage will be obtained.
• Formulations of the invention suitable for oral administration may be in the form of capsules (e.g. hard or soft shell gelatin capsule), cachets, pills, tablets, lozenges, powders, granules, pellets, dragees, e.g., coated (e.g., enteric coated) or uncoated, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles or as mouth washes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient.
• capsules e.g. hard or soft shell gelatin capsule
• cachets e.g., pills, tablets, lozenges
• powders granules, pellets, dragees, e.g., coated (e.g., enteric coated) or uncoated, or as a solution or a suspension
• a compound of the present invention may also be administered as a bolus, electuary or paste, or incorporated directly into the subject's diet.
• these pellets can be formulated to (a) provide for instant or rapid drug release (i.e., have no coating on them); (b) be coated, e.g., to provide for sustained drug release over time; or (c) be coated with an enteric coating for better gastrointestinal tolerability. Coating may be achieved by conventional methods, typically with pH or time-dependent coatings, such that the compound(s) of the invention is released in the vicinity of the desired location, or at various times to extend the desired action.
• Such dosage forms typically include, but are not limited to, one or more of cellulose acetate phthalate, polyvinylacetate phthalate, hydroxypropyl methyl cellulose phthalate, ethyl cellulose, waxes, and shellac.
• a compound of the present invention may be mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, or any of the following: fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, or silicic acid; binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose or acacia; humectants, such as glycerol; disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; solution retarding agents, such as paraffin; absorption accelerators, such as quaternary ammonium compounds; wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; absorbents, such as kaolin and bentonite clay; lubricants, such as talc, calcium
• compositions may also comprise buffering agents.
• Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.
• Peroral compositions typically include liquid solutions, emulsions, suspensions, and the like.
• the pharmaceutically acceptable vehicles suitable for preparation of such compositions are well known in the art.
• Typical components of carriers for syrups, elixirs, emulsions and suspensions include ethanol, glycerol, propylene glycol, polyethylene glycol, liquid sucrose, sorbitol and water.
• typical suspending agents include methyl cellulose, sodium carboxymethyl cellulose, tragacanth, and sodium alginate;
• typical wetting agents include lecithin and polysorbate 80; and typical preservatives include methyl paraben and sodium benzoate.
• Peroral liquid compositions may also contain one or more components such as sweeteners, flavoring agents and colorants disclosed above.
• compositions suitable for injectable use may include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions.
• the composition must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.
• Sterile injectable solutions can be prepared by incorporating the therapeutic agent in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization.
• dispersions are prepared by incorporating the therapeutic agent into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above.
• a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above.
• the methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient (i.e., the therapeutic agent) plus any additional desired ingredient from a previously sterile-filtered solution thereof.
• compositions are also provided which are suitable for administration as an aerosol, by inhalation. These formulations comprise a solution or suspension of the desired compound of any Formula herein or a plurality of solid particles of such compound(s).
• the desired formulation may be placed in a small chamber and nebulized. Nebulization may be accomplished by compressed air or by ultrasonic energy to form a plurality of liquid droplets or solid particles comprising the agents or salts.
• the liquid droplets or solid particles should have a particle size in the range of about 0.5 to about 5 microns.
• the solid particles can be obtained by processing the solid agent of any Formula described herein, or a salt thereof, in any appropriate manner known in the art, such as by micronization.
• the size of the solid particles or droplets will be, for example, from about 1 to about 2 microns.
• a pharmaceutical formulation suitable for administration as an aerosol may be in the form of a liquid, the formulation will comprise a water-soluble agent of any Formula described herein, or a salt thereof, in a carrier which comprises water.
• a surfactant may be present which lowers the surface tension of the formulation sufficiently to result in the formation of droplets within the desired size range when subjected to nebulization.
• compositions of this invention can also be administered topically to a subject, e.g., by the direct laying on or spreading of the composition on the epidermal or epithelial tissue of the subject, or transdermal ⁇ via a "patch".
• Such compositions include, for example, lotions, creams, solutions, gels and solids.
• These topical compositions may comprise an effective amount, usually at least about 0.1%, or even from about 1% to about 5%, of a compound of the invention.
• Suitable carriers for topical administration typically remain in place on the skin as a continuous film, and resist being removed by perspiration or immersion in water.
• the carrier is organic in nature and capable of having dispersed or dissolved therein the therapeutic agent.
• the carrier may include pharmaceutically acceptable emollients, emulsifiers, thickening agents, solvents and the like.
• compositions useful for attaining systemic delivery of the subject agents include sublingual, buccal and nasal dosage forms.
• Such compositions typically comprise one or more of soluble filler substances such as sucrose, sorbitol and mannitol; and binders such as acacia, microcrystalline cellulose, carboxymethyl cellulose and hydroxypropyl methyl cellulose. Glidants, lubricants, sweeteners, colorants, antioxidants and flavoring agents disclosed above may also be included.
• the compound(s) of the invention may also be administered parenterally, intraperitoneal ⁇ , intravenously, intraspinally, intrathecal ⁇ or intracerebrally.
• the compound(s) of the invention can be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms.
• the method of treatment of the present invention may also include co-administration of the at least one compound according to the invention, or a pharmaceutically acceptable salt thereof together with the administration of another therapeutically effective agent.
• an additional aspect of the invention relates to methods of concomitant therapeutic treatment of a subject, comprising administering to a subject in need thereof an effective amount of a first agent and a second agent, wherein the first agent is as defined in Formula I 1 IA, II, HA, III, or IHA and the second agent is for the prevention or treatment of any one of disorder or disease indicated hereinbefore.
• the term "concomitant” or “concomitantly” as in the phrases “concomitant therapeutic treatment” or “concomitantly with” includes administering a fist agent in the present of a second agent.
• a concomitant therapeutic treatment method includes methods in which the first, second, third or additional agents are co-administered.
• a concomitant therapeutic treatment method also includes methods in which the first or additional agents are administered in the presence of a second or additional agents, wherein the second or additional agents, for example, may have been previously administered.
• a concomitant therapeutic treatment method may be executed step-wise by different actors. For example, one actor may administer to a subject a first agent and as a second actor may administer to the subject a second agent and the administering steps may be executed at the same time, or nearly the same time, or at distant times, so long as the first agent (and/or additional agents) are after administration in the presence of the second agent (and/or additional agents).
• the actor and the subject may be the same entity (e.g. a human).
• the invention also relates to a method for preventing, reducing or eliminating a symptom or complication of any one of the above mentioned disease or condition.
• the method comprises administering, to a subject in need thereof, a first pharmaceutical composition comprising at least one compound of the invention and a second pharmaceutical composition comprising one or more additional active ingredients, wherein all active ingredients are administered in an amount sufficient to inhibit, reduce, or eliminate one or more symptoms or complications of the disease or condition to be treated.
• the administration of the first and second pharmaceutical composition is temporally spaced apart by at least about two minutes.
• the first agent is a compound of Formula I 1 IA, II, MA, III, or IHA as defined herein, or a pharmaceutically acceptable salt thereof.
• the second agent may be selected from the following list of compounds: Z-Asp-Phg-Val- Aspmethyl vinyl sulfone (53) F) Screening assays
• the compounds of the present invention may also be used in screening methods. For instance, these compounds may be used in methods for tracking activity of caspases in vitro and/or in vivo. The compounds of the present invention may also be helpful for identifying other compounds that bind to a caspase active side.
• the compounds of the invention are labeled or tagged (e.g. fluorescently or radioactively labeled, affinity tag). Fluorescent or radiolabeled compounds may also be useful in diagnostic assays.
• the caspase is bound to a support, and a labeled compound of the invention is added to the assay.
• the compound of the invention may be bound to the support and the caspase is added.
• drug candidate or test compounds
• test compounds are used interchangeably and describe any molecule, for example, protein, oligopeptide, small organic molecule, polysaccharide, polynucleotide, and the like, to be tested for bioactivity.
• the signals that are detected in the assay may include fluorescence, resonance energy transfer, time resolved fluorescence, radioactivity, fluorescence polarization, plasma resonance, or chemiluminescence and the like, depending on the nature of the label.
• Detectable labels useful in performing screening assays in this invention include a fluorescent label such as Fluorescein, Oregon green, dansyl, rhodamine, tetramethyl rhodamine, texas red, Eu 3+ ; a chemiluminescent label such as luciferase; colorimetric labels; enzymatic markers; or radioisotopes such as tritium, I 125 and the like.
• Affinity tags, which may be useful in performing the screening assays of the present invention include be biotin, polyhistidine and the like.
• the compound(s) of the invention may be packaged as part of a kit, optionally including a container (e.g. packaging, a box, a vial, etc).
• the kit may be commercially used according to the methods described herein and may include instructions for use in a method of the invention.
• Additional kit components may include acids, bases, buffering agents, inorganic salts, solvents, antioxidants, preservatives, or metal chelators.
• the additional kit components are present as pure compositions, or as aqueous or organic solutions that incorporate one or more additional kit components. Any or all of the kit components optionally further comprise buffers.
• the compound(s) of the invention may or may not be administered to a patient at the same time or by the same route of administration. Therefore, the methods of the invention encompass kits which, when used by the medical practitioner, can simplify the administration of appropriate amounts of two or more active ingredients to a patient.
• a typical kit of the invention comprises a unit dosage form of a at least one compound according to the invention, e.g., a compound of Formula I, IA, II, HA, III, or IHA as defined herein or a pharmaceutically acceptable salt thereof, and a unit dosage form of at least one additional active ingredient.
• additional active ingredients include, but are not limited to any of the compounds that could be used in combination with the compound(s) of the invention as indicated herein before.
• Kits of the invention can further comprise devices that are used to administer the active ingredients.
• devices include, but are not limited to, syringes, drip bags, patches, inhalers, enemas, and dispensers for the administration of suppository formulations.
• Kits of the invention can further comprise pharmaceutically acceptable vehicles that can be used to administer one or more active ingredients.
• the kit can comprise a sealed container of a suitable vehicle in which the active ingredient can be dissolved to form a particulate-free sterile solution that is suitable for parenteral administration.
• suitable vehicles are provided hereinbefore.
• Fmoc-Ala (2'-quinolyl)-OH (0.152 g, 0.347 mmol) was solvated in DMF (1 mL) and CH 2 CI 2 (0.9 mL).
• L-VaI allylester toluene-4-sulfonate (0.115 g, 1.01 eq) in 0.3 mL of CH 2 CI 2 was aded, followed with 4-methylmorpholine (0.04 mL, 1.05 eq) and EDC (0.0681g, 1.02eq). The mixture was stirred for 3 hours, and then it was extracted using CH 2 CI 2 / brine. The organic layer was dried over MgSO 4 , filtered off and concentrated to dryness.
• Ts-Ala(2'-quinolyl)-OAIIyl (0.010 g, 0.02 mmol) was dissolved in THF (3.5 ml.) and rotary evaporated to dryness, the sample was then redissolved in THF (3.5 mL) and vacuum aspirated (3 * 1 min of aspiration), followed by replacement of the atmosphere with Argon.
• Pd(PPh 3 ) 4 (0.0036 g, 0.16 eq) was added under Argon, followed by Morpholine (0.007ml_, 4.1 eq).
• the sample flask was then covered with a tin foil and kept under stirring for 3 days under Argon.
• the compound was evaporated to dryness and the obtained residue was subjected to purification on C 18 using a gradient: MeOH/H 2 O (15 to 100 %) to get 0.006 g of the desired compound.
• Fmoc lndanylglycine (0.54 g, 1.315 mmol) was solvated in CH 2 CI 2 (5 mL) and DMF (1.8 ml.) followed with the addition of L- VaI allylester (0.437 g, 1 eq), 4-methylmorpholine (0.15 ml, 1.04 eq) and 4 min latter DMAP (14.5 mg, 0.09 eq) then EDC (0.265 g, 1.05 eq). The mixture was stirred for 1 hour 45 min. Then it was extracted using EtOAc/ brine. The organic layer was dried over MgSO 4 , filtered off and concentrated to dryness.
• Fmoc- Indanylglycine-Val-OAIIyl (0.758 g, 1.31 mmol) was solvated in CH 2 CI 2 (5ml_) and DMF (1.8 mL) followed by dropwise addition of piperidine (1.05 mL, 8.1 eq) over 30 seconds. After 45 min, the mixture was subjected to extraction CH 2 CI 2 / brine (30/15 mL) and saturated NH 4 CI (5 mL). The organic layer was dried over MgSO 4 , concentrates and purified by silica using a gradient of MeOH/ CH 2 CI 2 (0 to 5 %) to get 0.45 g of the desired compound.
• Indanylglycine-Val-OAIIyl (0.46 g, 1.3 mmol) and Z-L-Asp(OfBu)-OH (0.425 g, 1.3 mmol) were solvated in CH 2 CI 2 (4.5 mL) followed by addition of 4-methylmorpholine (1.45 ml_, 1.01 eq), DMAP (14 mg, 0.09 eq) then EDC (0.251 g, 1.01 eq). The mixture was stirred for 1 hour 40 min. Then it was extracted using CH 2 CI 2 / brine. The organic layer was dried over MgSO 4 , concentrates and purified by silica using a gradient of EtOAc/Hexane (10 to 80 %) to get 0.578 g of the desired compound.
• Z-Asp( ⁇ -tert-butyl)-Phg-Val-OAIIyl (0.064 g; 0.107 mmol) was dissolved under argon in dry THF (3 ml, inhibitor free). The solvent was degassed three times under argon before adding morpholine (28 ul, 3 eq), followed by Tetrakis (13 mg). The mixture was stirred for 3.5 days at room temperature. The mixture was then concentrated under vacuum (12 mbar) and purified on silica gel (gradient methanol/dichloromethane: 1 to 14 %) to afford 46 mg of the desired Z-Asp( ⁇ -tert-Butyl)-Phg-Val-OH.
• Fmoc- GIu(O-(Bu)-VaI(OAIIyI) (0.394 g, 0.698 mmol) was solvated in CH 2 CI 2 (4 mL) followed by addition of piperidine (0.550 mL, 7.98 eq). After 40 min, the mixture was evaporated to dryness and co-evaporated with CH 2 CI 2 (20 ml_ * 2) followed with high vacuum for 10 min to remove the exces of piperidine. The sample was purified on silica with a gradient of MeOH /CH 2 CI 2 (0 to 7 %) to get 0.191 g of the desired compound.
• GIu-VaI (OAIIyI) (0.189 g, 0.55 mmol) was solvated in CH 2 CI 2 (2.5 ml) followed addition of Z-Asp (OfBu)-OH (0.187 g, 1.05 eq), DMAP (5.98 mg, 0.088 eq) and 4- methylmorpholine (0.065 ml, 1.07 eq), then EDC (0.109 g, 1.03 eq.
• the vial was rinced with 0.5 ml of dichloromethane). The mixture was stirred for 2 hour at room temperature. Then it was extracted using CH 2 CI 2 / brine. The organic layer was dried over MgSO 4 , filtered off and concentrated. The obtained residue was purified on silica using a gradient EtOAc/Hex (10 to 100 %) to get 0.300 g of the desired compound.
• the vial was rinsed with THF (0.3 mL*2) and added to the reaction mixture. After 45 min of stirring, the solution was quenched with a solution of saturated ammonium chloride (5 mL) and extracted with EtOAc (30 mL). The organic layer was washed with brine, dried over MgSO 4 , filtered and concentrated. The residue was purified on silica using a gradient of EtOAc/Hex (0 to 40 %) to get first the cis isomer and then 0.183 g of the trans isomer compound.
• Boc-Asp ( ⁇ -tert-butyl) Chlorovinyl-methyl vinyl sulfone (0.182 g, 0.4937 mmol) was solvated in CH 2 CI 2 (0.7 mL) followed by the addition of Et 2 O (0.7 mL).
• p-Toluene sulfonic Acid monohydrate (0.101 g, 1.07 eq) was added in one shot. After 15 hours of stirring at room temperature, it was then diluted with Ether (8 mL) and filtered off. The while solid was then dried over vacuum, 0.12 g of the desired compound was obtained.
• Example 8 Synthesis of compound 30 (Asp( ⁇ -Methyl) methyl vinyl sulfone tosyl salt) a) Boc-Aspart ⁇ mol( ⁇ -Methyl)
• Boc Asp (OCH 3 )-CH 2 OH in CH 2 CI 2 (2.6 mL) was added dropwise over a period of 20 min and the reaction was stirred for an extra 15 min at -65 0 C.
• Boc-Asp( ⁇ -Methyl) methyl vinyl sulfone (0.172 g, 0.56 mmol) was solvated in CH 2 CI 2 (0.44 mL) followed by the addition of Et 2 O (0.44 ml_).
• p-Toluene sulfonic Acid monohydrate (0.19 g, 1.04 eq) was added in one shot. After 15 hours of stirring at room temperature, it was then diluted with Ether (2 mL) and filtered off. The while solid was then dried over vacuum, 0.14O g of the desired compound was obtained.
• Boc-Asp( ⁇ -tert-butyl) methyl vinyl sulfone (0.158 g, 0.4534 mmol) was solvated in CH 2 CI 2 (0.7 mL) followed by the addition of Et 2 O (0.7 mL).
• p-Toluene sulfonic Acid monohydrate (0.0878 g, 1.02 eq) was added in one shot.
• the use of excess of PTSA hydrate cleaves both boc and tert-butyl groups unlike what was reported by Palmer. After 15 hours of stirring at room temperature, it was then diluted with Ether (5 mL) and filtered off. The while solid was then dried over vacuum, 0.121 g of the desired compound was obtained.
• phenyl vinyl sulfone was obtained in one step from benzenesulfonyl fluoride and triethyl phosphorane in the presence of lithium hexamethyldisilazide at -78 0 C to get diethyl phenylsulfonylmethylphosphonate as described in the literature (Won Bum Jang et al., 1998).
• Boc-Asp( ⁇ -tert-Butyl)-vinyl phenyl sulfone (Gang Wang et al., 2003; Marion G. Gotz et al., 2004; Palmer, James T et al., 1995).
• Boc-L-Asp( ⁇ -tert-Butyl)-Vinyl phenyl sulfone (0.1 g, 0.243 mmol) was dissolved in a mix of dichloromethane and ether (0.7 / 0.7 ml), then PTSA hydrate (1eq) was added.
• PTSA hydrate (1eq) was added.
• the use of excess of PTSA hydrate cleaves both boc and tert-butyl groups unlike what was reported by Palmer.
• the mixture was stirred at room temperature overnight. Then, it was diluted with ether (8 ml). The white precipitate was filtered off and dried to yield to the desired compound as a white powder.
• the vial was rinsed with THF (1 mL) and added to the reaction mixture. After 2 hours of stirring at room temperature, the solution was quenched with a solution of ammonium chloride saturated (5 mL) and extracted with EtOAc (30 mL). The organic layer was washed with brine, dried over MgSO 4 , filtered off and concentrated. The residue was purified on silica using a gradient of EtOAc/Hexane (0 to 30%, 30 to 80 %) to get 0.359 g of the desired compound.
• Asp( ⁇ -tert-butyl) phenoxy vinyl sulfone (0.187 g, 0.444 mmol) was solvated in CH 2 CI 2 (OJmL) followed by the addition of Et 2 O (0.7 ml_).
• p-Toluene sulfonic Acid monohydrate (0.084 g, 1.01 eq) was added in one shot. After 15 hours of stirring at room temperature, it was then diluted with Ether (8 mL) and filtered off. The while solid was then dried over vacuum to get the desired compound.
• Chloromethyl isopropylsulfide (12.54 g, 100.6 mmol) was heated to 110 0 C follow by dropwise addition of triethylphosphonyl (21 mL, 1.10 eq). After stirring for 8 hours, the reaction was allowed to reach to room temperature. The sample was purified by distillation (110 0 C / 6 mmbar) to get 4.7 g of Diethyl (isopropylthio)methylphosphonate.
• Boc-Asp( ⁇ -tert-butyl) isopropyl vinyl sulfone (0.028 g, 0.0741 mmol) was solvated in CH 2 CI 2 (0.2ml_) followed by the addition of Et 2 O (0.2 ml_).
• p-Toluene sulfonic Acid monohydrate (0.0154 g, 1.09 eq) was added in one shot. After 15 hours of stirring at room temperature, it was then diluted with Ether (8 ml_) and filtered off. The while solid was then dried over vacuum, 11 mg of the desired compound was obtained.
• Boc-Asp( ⁇ -tert-butyl) morpholine vinyl sulfone (0.075 g, 0.178 mmol) was solvated in CH 2 CI 2 (0.2 mL) followed by the addition of Et 2 O (0.2 mL).
• p-Toluene sulfonic Acid monohydrate (0.034 g, 1.0 eq) was added in one shot. After 15 hours of stirring at room temperature, it was then diluted with Ether (0.5 mL) and filtered off. The while solid was then dried over vacuum, 0.045 g of the desired compound was obtained.
• the Z-Asp( ⁇ -tert-Butyl)-Ala(2'-quinolyl)-Val-OH (18 mg, 0.029 mmol) is dissolved in a mix of dichloromethane and DMF (0.39 ml / 0.13 ml). The mixture was allowed to reach - 15/-20 0 C before adding N-methyl morpholine (8 ⁇ l) followed 3 min latter with isobutyl chloroformate (7 ⁇ l). The mixture was stirred at -15 0 C for 10 minutes. Next, Asp ( ⁇ -tert- butyl) ⁇ chloromethyl-methylsulfone tosyl salt (13 mg, 1eq) was added in one shot, followed by N-methyl morpholine (8 ⁇ l).
• the Ts-AIa (2'-quinolyl)-OH (5.6 mg, 0.0123 mmol) was dissolved in a mix of CH 2 CI 2 / DMF (0.15/ 0.13 ml_). The mixture was allowed to reach - 15 °C / - 20 0 C (ice MeOH bath) before adding NMM (0.004 ml_) followed 3 min later with isobutylchloroformate (0.004 ml_). The mixture was stirred for 10 min at -15 °C before adding Asp ( ⁇ -tert-butyl) Chlorovinyl-methyl vinyl sulfone tosyl salt (6.3 mg, 1 eq) in one shot, followed with A- methylmorpholine (0.004 ml_).
• the mixture was sirred at - 15 0 C / -20 0 C for 30 min, it was then diluted with CH 2 CI 2 (5 ml_), then water (2 ml.) and the mixture was allowed to reach room temperature, extracted. The organic layer was dried over MgSO 4 , concentrated and purified by silica using Hex/ EtOAc (15 to 100%) to get 6 mg of the desired compound.
• Example 21 Synthesis of compound 50 (Ts-Ala(2'quinolyl)-Val-Asp- ⁇ chlorovinyl methylsulfone) Ts-Ala(2'quinolyl)-Val-Asp( ⁇ -tert-Butyl)- ⁇ chlorovinyl methylsulfone (6 mg) was dissolved in dichloromethane (0.15 ml), followed by addition of trifluoroacetic acid (0.2 ml). The mixture was stirred at room temperature overnight (15 h). It was then quenched with diethyl ether (5 ml) and the solvent was removed under vacuum. The obtained residue was diluted again with ether and the process was repeated twice. The precipitate was washed with 2* 1 ml of ether, dried to give 4 mg of the desired compound.
• the Z-Asp( ⁇ -methyl)-lndanylglycine-Val-OH (16.7 mg, 0.0301 mmol) is dissolved in a mix of THF and DMF (0.5 ml / 0.1 ml). The mixture was allowed to reach -15/-20 0 C before adding N-methyl morpholine (9 ⁇ l) followed 3 min latter with isobutyl chloroformate (8 ⁇ l). The mixture was stirred at -15 0 C for 10 minutes.
• the Z-Asp( ⁇ -tert-Butyl)-Phg-Val-OH (17 mg, 0.0306 mmol) is dissolved in a mix of dichloromethane and DMF (0.39 ml / 0.13 ml). The mixture was allowed to reach -15/-20 0 C before adding N-methyl morpholine (8 ⁇ l) followed 3 min latter with isobutyl chloroformate (8 ⁇ l). The mixture was stirred at -15 0 C for 10 minutes. Next, Asp ( ⁇ -tert- butyl) methyl vinyl sulfone tosyl salt (12 mg, 1 eq) was added in one shot, followed by N- methyl morpholine (8 ⁇ l).
• the Z-Asp( ⁇ -tert-Butyl)- AI(2'-quinolyl)-Val-OH (19 mg, 0.0306 mmol) is dissolved in a mix of dichloromethane and DMF (0.4 ml / 0.14 ml). The mixture was allowed to reach - 15/-20 0 C before adding N-methyl morpholine (9 ⁇ l) followed 3 min latter with isobutyl chloroformate (9 ⁇ l). The mixture was stirred at -15 0 C for 10 minutes. Next, Asp ( ⁇ -tert- butyl) methyl vinyl sulfone tosyl salt (13.1 mg, 1 eq) was added in one shot, followed by N-methyl morpholine (9 ⁇ l).
• the Z-Asp( ⁇ -tert-Butyl)-lndanylglycine -VaI-OH (21 mg, 0.0353 mmol) is dissolved in a mix of dichloromethane and DMF (0.44 ml / 0.15 ml). The mixture was allowed to reach - 15/-20 0 C before adding N-methyl morpholine (10 ⁇ l) followed by isobutyl chloroformate (9 ⁇ l). The mixture was stirred at -15 0 C for 10 minutes. Next, Asp ( ⁇ -tert-butyl) methyl vinyl sulfone tosyl salt (15 mg, 1 eq) was added in one shot, followed by N-methyl morpholine (10 ⁇ l).
• the Z-Asp( ⁇ -tert-Butyl)-Glu( ⁇ -tert-Butyl)-Val-OH (18 mg, 0.029 mmol) is dissolved in a mix of dichloromethane and DMF (0.39 ml / 0.13 ml). The mixture was allowed to reach - 15/-20 0 C before adding N-methyl morpholine (8 ⁇ l) followed 3 min latter with isobutyl chloroformate (7 ⁇ l). The mixture was stirred at -15 0 C for 10 minutes. Next, Asp ( ⁇ -tert- butyl) methyl vinyl sulfone tosyl salt (13mg) was added in one shot, followed by N-methyl morpholine (8 ⁇ l).
• the Z-VaI-OH (15 mg, 0.059 mmol) is dissolved in a mix of dichloromethane and DMF (0.5 ml / 0.2 ml). The mixture was allowed to reach -15/-20 0 C before adding N-methyl morpholine (10 ⁇ l) followed by isobutyl chloroformate (9 ⁇ l). The mixture was stirred at - 15 0 C for 10 minutes. Next, Asp ( ⁇ -tert-butyl) methyl vinyl sulfone tosyl salt (25mg, 1eq) was added in one shot, followed by N-methyl morpholine (10 ⁇ l).
• Example 36 Synthesis of compound 62 (Z-Asp( ⁇ -tert-Butyl)-Ala(2'-quinolyl)-Val- Asp( ⁇ -tert-Butyl)phenyl vinyl sulfone)
• the Z-Asp( ⁇ -tert-Butyl)-Ala(2'-quinolyl)-Val-OH (19 mg, 0.0306 mmol) is dissolved in a mix of dichloromethane and DMF (0.39 ml / 0.13 ml). The mixture was allowed to reach - 15/-20 0 C before adding N-methyl morpholine (8 ⁇ l) followed by isobutyl chloroformate (7 ⁇ l). The mixture was stirred at -15 0 C for 10 minutes. Next, Asp ( ⁇ -tert-butyl) phenyl vinyl sulfone tosyl salt (11 mg) was added in one shot, followed by N-methyl morpholine (8 ⁇ l).
• the Z-Asp( ⁇ -tert-Butyl)-AI(2'-quinolyl)-Val-OH (10 mg, 0.016 mmol) is dissolved in a mix of dichloromethane and DMF (0.24 ml / 0.080 ml). The mixture was allowed to reach - 15/-20 0 C before adding N-methyl morpholine (5 ⁇ l) followed 3 min latter with isobutyl chloroformate (5 ⁇ l). The mixture was stirred at -15 0 C for 10 minutes.
• Example 39 Synthesis if compound 65 (Z-Asp-Ala(2'-quinolyl)-Val-Aspphenoxy vinyl sulfone)
• the mixture was stirred 35 minutes at -15 /-20 0 C and then diluted with 5 ml of dichloromethane and quenched with 1.5 ml of a saturated solution of sodium bicarbonate. After that, the organic layer was washed with sodium bicarbonate and dried over magnesium sulphate. The solvent was evaporated to dryness. The obtained residue was purified on silica gel (gradient: ethyl acetate / hexane: 12 to 100%) to afford 10 mg of the desired compound.
• This compound was synthesized from Asp( ⁇ -tert-butyl)isopropyl vinyl sulfone tosyl salt (43) and Cbz-Asp(O-fl3u)lndanylglycine-Val-OH (12) via anhydride mixte method as for Z-Asp-lndanylglycine-Val- Aspmethyl vinyl sulfone (57).
• This compound was synthesized as the corresponding Z-Asp-Ala(2'-quinolyl)-Val- Asppheny) vinyl sulfone (63).
• This compound was synthesized from Z-Asp-Tyr(OtBu)-Val-OH (75) and Asp(OtBu)- Vinyl methy sulfone tosyl salt (33) via anhydride mixte method as for Z-Asp-Phg-Val- Aspmethyl vinyl sulfone (53).
• Z-Asp-Tyr(OtBu)-Val-OH was synthesized as Cbz-Asp(O-tBu)-Glu(O-tBu)-Val-OH (20).
• This compound was synthesized from Z-Tyr(OtBu)-Glu(OtBu)-Val-OH (80) and Asp(OtBu)-Vinyl methy sulfone tosyl salt (33) via anhydride mixte method as for Z-Asp- Phg-Val-Aspmethyl vinyl sulfone (53).
• Z-Tyr(OtBu)-Glu(OtBu)-Val-OH (80) was synthesized as Cbz-Asp(O-tBu)-Glu(O-tBu)- VaI-OH (20).
• This compound was synthesized from Z-Asp(0tBu)-Ala(2'-pyridyl)-Val-0H (83) and Asp(OtBu)-Vinyl phenyl sulfone tosyl salt (37) via anhydride mixte method as for Z-Asp- Ala(2'-quinolyl)-Val-Aspphenyl vinyl sulfone (63).
• Example 50 Selectivity of exemplary compounds for caspase-3 relative to caspase-1, caspase-5, caspase-7, and caspase-9
• Compound 55 showed an inhibitory effect on both caspase-3 and - 7 activity. However, based on the IC50 values calculated, it was about 200 fold more selective of Caspase-3 over Caspase-7 (see Table 2). No significant inhibitory activity of the compound was observed for caspase-1 , -5, and -9 at the tested dose-range. Inhibition of Caspase-1 , -5, and -9 was achieved (25%, 21%, and 57% respectively) but at extremely high concentration (about 10,00OnM).
• Compound 63 Similar to Compound 55, Compound 63 also showed high selectivity in caspase-3 activity inhibition relative to caspase-1 , -5, -7, and -9. Compound 63 also showed an inhibitory effect on caspase-3 and -7, with about a 50 fold selectivity of caspase-3 over caspase-7. No significant inhibitory activity was observed for caspase-1 , -5, and -9 at the tested dose-range of Compound 63. The data indicates that both Compound 55 and Compound 63 are highly potent and selective compounds for caspase-3 activity inhibition.
• Compound 48 was able to inhibit specific groups of caspases.
• the compound was able to inhibit caspase-3, -7, and -9 with IC50 values of about 8-30 nM, 0.4-0.9 uM and 1-1.8 uM respectively.
• Compound 48 shows inhibition of Group III caspases (caspase-3 and -7) with high potency, and inhibition of Group Il caspase (caspase-9) with a lower yet significant efficacy.
• Compound 57 is a potent inhibitor of Group III caspases (caspase-3 and -7).
• Compound 57 inhibited caspase-3 and -7 activities with IC50 values of about 30-90 nM and about 180-300 nM respectively. Therefore, data indicates that Compound 48 and Compound 57 are not specific inhibitors for caspase-3 activity, but are in fact inhibitors targeted at specific groups of caspases.
• Compound 88 has a dual inhibition effect on both caspase-1 and caspase-3 activity. It is a potent inhibitor of caspase-3 activity, with an IC50 value of about 30-90 nM. It also inhibits caspase-1 activity with an IC50 value of about 0.6-1.2 uM.
• Example 51 In vitro inhibitory activity of Caspase-3 inhibitors
• HeIa cells are seeded in 96 well plates and incubated for 4 hours with staurosporine, a well characterized inducer of apoptosis, alone or together with different concentrations of compound (50, 25, 10 and 3uM). After formaldehyde-based fixation, the cells are stained with a fluorescein-labeled anti-cleaved PARP antibody (Cell signaling, Cat#: 9547) and counterstained with Hoechst33342 (Invitrogen, Cat#: H3570) to mark all nuclei.
• a fluorescein-labeled anti-cleaved PARP antibody Cell signaling, Cat#: 9547
• Hoechst33342 Invitrogen, Cat#: H3570
• Fluorescence images are taken on a CellomicsTM microscope system (Thermo Scientific, Pittsburgh, USA) with the Hoechst stain in the blue channel and the cleaved PARP antibody stain in the green channel.
• the percentage of cleaved PARP positive cells is determined by calculating the ratio between nuclei with a cleaved PARP antibody staining above a certain threshold and all (Hoechst positive) nuclei.
• the efficacy of caspase-3 inhibition is determined by calculating the ratio between cleaved PARP positive cells after staurosporine incubation together with compounds and staurosporine incubation without compounds. Results are summarized in Table 2.
• results of this assay show that compounds that inhibited Caspase- 3 activity in the enzymatic assay with an IC50 below 10OnM were generally also effective in inhibiting PARP cleavage in vitro, although a major factor contributing to a compound's activity / effectivity in this assay is the compound's cell membrane permeability coefficient (the higher the permeability coefficient, the greater the amount of the compound in the media reaching inside the cell).
• Certain modifications in the molecular composition of the compounds improved the inhibitory activity further; for example Compound 51, Compound 53, Compound 57, and Compound 76 reduced the percentage of PARP positive cells after staurosporine treatment by more than 50%, with Compound 51 and Compound 57 being the most effective compounds reaching values of about 65% inhibition, and Compound 53 also being highly effective relative to DEVD- fmk (positive control compound with low caspase-3 selectivity but highly cell membrane permeable with general caspase inhibitory activity), and Compounds 48, 55, and 59 also showing effectivity especially relative to DEVD-fmk.
• DEVD- fmk positive control compound with low caspase-3 selectivity but highly cell membrane permeable with general caspase inhibitory activity
• Compounds 48, 55, and 59 also showing effectivity especially relative to DEVD-fmk.

Landscapes

• Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Life Sciences & Earth Sciences (AREA)
• General Health & Medical Sciences (AREA)
• Medicinal Chemistry (AREA)
• Chemical Kinetics & Catalysis (AREA)
• General Chemical & Material Sciences (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Pharmacology & Pharmacy (AREA)
• Animal Behavior & Ethology (AREA)
• Public Health (AREA)
• Veterinary Medicine (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Molecular Biology (AREA)
• Biochemistry (AREA)
• Biophysics (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Genetics & Genomics (AREA)
• Neurology (AREA)
• Neurosurgery (AREA)
• Oncology (AREA)
• Biomedical Technology (AREA)
• Virology (AREA)
• Immunology (AREA)
• Physical Education & Sports Medicine (AREA)
• Communicable Diseases (AREA)
• Diabetes (AREA)
• Orthopedic Medicine & Surgery (AREA)
• Hematology (AREA)
• Rheumatology (AREA)
• Cardiology (AREA)
• Gastroenterology & Hepatology (AREA)
• Pulmonology (AREA)
• Heart & Thoracic Surgery (AREA)
• Pain & Pain Management (AREA)
• Crystallography & Structural Chemistry (AREA)
• Psychology (AREA)
• Urology & Nephrology (AREA)

Priority Applications (13)

Applications Claiming Priority (2)

Related Child Applications (2)

Publications (1)

Family

ID=41339699

Family Applications (1)

Country Status (11)

Cited By (4)

Families Citing this family (7)

Citations (8)

Family Cites Families (29)

• 2009
  • 2009-05-21 EP EP09749369.6A patent/EP2288615B1/en not_active Not-in-force
  • 2009-05-21 ES ES09749369.6T patent/ES2640814T3/es active Active
  • 2009-05-21 WO PCT/CA2009/000696 patent/WO2009140765A1/en not_active Ceased
  • 2009-05-21 AU AU2009250295A patent/AU2009250295B2/en not_active Ceased
  • 2009-05-21 CA CA2724785A patent/CA2724785C/en active Active
  • 2009-05-21 JP JP2011509829A patent/JP2011524340A/ja not_active Ceased
  • 2009-05-21 SG SG2013038997A patent/SG191591A1/en unknown
  • 2009-05-21 CN CN200980128709.7A patent/CN102105485B/zh not_active Expired - Fee Related
  • 2009-05-21 US US12/993,565 patent/US8791235B2/en active Active
  • 2009-05-21 DK DK09749369.6T patent/DK2288615T3/en active
• 2010
  • 2010-11-18 IL IL209407A patent/IL209407A0/en active IP Right Grant
• 2014
  • 2014-06-17 US US14/307,025 patent/US9562069B2/en active Active
  • 2014-12-26 JP JP2014265816A patent/JP5995951B2/ja not_active Expired - Fee Related
• 2016
  • 2016-08-23 JP JP2016162904A patent/JP2017039723A/ja active Pending
• 2017
  • 2017-01-06 US US15/400,323 patent/US10167313B2/en active Active

Patent Citations (9)

Non-Patent Citations (11)

Also Published As

Similar Documents

Legal Events