Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
  • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
  • A61K51/04—Organic compounds
  • A61K51/041—Heterocyclic compounds
  • A61K51/044—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins
  • A61K51/0446—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
  • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
  • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
  • A61K47/545—Heterocyclic compounds

Definitions

• This invention relates to novel therapeutic agents that bind to 5-HT receptors and modulate their role in living systems. More particularly, the invention relates to novel therapeutic agents that bind to 5-HT receptors and modulate their activity by acting as a multibinding agent.
• the multibinding agents ofthe invention comprise from 2-10 ligands covalently connected by a linker or linkers, wherein said ligands in their monovalent (i.e. unlinked) state are capable of binding to a 5-HT receptor and modulating its activity.
• the manner in which the ligands are linked is such that the multibinding agents so constructed demonstrate an increased biological and or therapeutic effect as compared to the same number of unlinked ligands available for binding to the 5-HT receptor.
• the compounds ofthe invention are particularly useful in treating conditions in a mammal that are mediated by 5-HT receptors. Accordingly, the invention also relates to pharmaceutical compositions comprising a pharmaceutically acceptable excipient and an effective amount of a compound ofthe invention, and to methods of using such compounds and pharmaceutical compositions containing them for the treatment of such conditions.
• the invention relates to methods of preparing such compounds.
• Serotonin (5-hydroxytryptamine) is a major neurotransmitter in mammals, and plays a role that is central to the function of the central and peripheral nervous system. It is responsible for controlling important physiological functions such as sleep, appetite, pain, movement, and temperature regulation. Accordingly, much emphasis has been placed on modulating the action of serotonin for treating disease states such as migraine, headache, itch, motion sickness, depression, emesis, memory loss, anxiolytic disorders, obesity, gastrointestinal disorders, irritable bowel syndrome, and the like.
• Serotonin is the endogenous ligand for a variety of cell surface receptors. These include the seven transmembrane G-protein coupled receptors (GPCR), receptor ligand gated ion channels (e.g. 5HT3), and the twelve transmembrane serotonin reuptake protein.
• GPCR transmembrane G-protein coupled receptors
• 5HT3 receptor ligand gated ion channels
• 12 transmembrane serotonin reuptake protein the twelve transmembrane serotonin reuptake protein.
• 5HT receptor subtypes there are believed to be at least 14 mammalian 5HT receptor subtypes. These receptors may be positively coupled to adenylate cyclase (5HT4, 5HT6, 5HT7) or negatively coupled to adenylate cyclase (5HT1A, 5HT1B, 5HT1D, 5HT1F) or coupled to phospholipase C (5HT2A, 5HT2B, 5HT2C). The effector system for the remaining subtypes, 5HT5A and 5HT5B has not been determined.
• the 5HT1 class is negatively coupled to adenlyate cyclase through Gi (see, for example, Saxena, Pramod R. Modern 5-HT receptor classification and 5-HT based drugs. Expert Opin. Invest. Drugs (1994), 3(5), 513-23).
• triptan class are small molecule agonists (or partial agonists) for seven transmembrane cell surface receptors. They are believed to act both through peripheral and central mechanisms.
• the triptan class of drugs shows mixed subtype activity across a minimum of four serotonin subtypes.
• it is the agonism of the 5HT1 receptor, in particular the 5HT1B, 5HT1D, and 5HT1F receptors, that is proposed to be responsible for the efficacy of triptans in the treatment of migraine.
• drugs that act indiscriminately upon two or more of these subtypes i.e. are not selective for the targeted subtype
• are reported to have undesirable side effects e.g. cardiovascular contraindications, including chest tightness and pressure.
• a drug that is selective for the 5HT1D receptor may reduce these side effects.
• the present invention relates to novel multibinding agents wherein a multibinding agent comprises 2-10 ligands. which may be the same or different, covalently connected by a linker or linkers, which may be the same or different, each of said ligands comprising a ligand domain capable of binding to a 5-HT receptor.
• the preferred multibinding agents are represented by Formula I:
• each of said ligands comprises a ligand domain state capable of binding to a 5- HT receptor.
• q is less than p.
• the invention in a second aspect, relates to a method of modulating 5-HT receptors in a biologic tissue, preferably in a mammalian or avian subject, comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound of Formula I, or a salt thereof.
• the invention in a third aspect, relates to a pharmaceutical composition comprising a therapeutically effective amount of one or more compounds of Formula I, or a pharmaceutically available salt thereof, admixed with at least one pharmaceutically acceptable excipient.
• the invention relates to processes for preparing the compounds of Formula I.
• the invention in a fifth aspect, relates to a method for identifying a multibinding agent capable of binding to a 5-HT receptor, comprising preparing an array of multimeric agents, contacting the multimeric agent array with a 5-HT receptor, and selecting a multibinding agent based upon its ability to bind to the 5-HT receptor.
• Bioactive ligands in general involve molecular interactions between bioactive ligands and their receptors, in which the receptor "recognizes " ' a molecule (a ligand) or portion of a molecule (a ligand domain).
• a ligand a molecule
• a ligand domain a molecule
• 5-HT receptors drugs
• the result of this interaction can be either to initiate the desired biological effect of the 5-HT receptor, or alternatively to inhibit or alter (i.e. to modulate) the normal function of the 5-HT receptor. Accordingly, the modulation of such processes is regarded as an important target for drug development.
• the interaction of a 5-HT receptor with a ligand may be described in terms of
• affinity and “specificity”. The affinity and specificity of any given ligand receptor interaction are dependent upon the complementarity of molecular binding surfaces and the energetic costs of complexation. Affinity may be quantified by the equilibrium constant of complex formation. Specificity relates to the difference in affinity between different ligands binding to the same receptor subtype, or the same ligand binding to different receptor subtypes.
• the compounds of the invention are multibinding agents, and although not wishing to be bound or restricted by any particular theory or proposed mechanism of action, it is believed that the surprising activity of these compounds at least in part arises from their ability to bind in a multivalent manner with 5-HT receptors (i.e. the ligand binding site), and thus lower the energetic costs of binding.
• Multivalent binding interactions are characterized by the concurrent interaction of at least two ligands of a multibinding agent with multiple ligand binding sites, which may be multiple distinct 5- HT receptors or multiple distinct binding sites on a single 5-HT receptor. Multivalent interactions differ from collections of individual monovalent interactions in that they give rise to an enhanced biological effect.
• alkyl refers to a monoradical branched or unbranched saturated hydrocarbon chain, preferably having from 1 to 40 carbon atoms, preferably 1-10 carbon atoms, more preferably 1-6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n- butyl, secondary butyl, tert-butyl, n-hexyl, n-octyl, n-decyl, n-dodecyl, 2-ethyldodecyl, tetradecyl, and the like, unless otherwise indicated.
• lower alkyl referes to an alkyl group as defined above having from 1-6 carbon atoms.
• substituted alkyl refers to an alkyl group as defined above having from 1 to 5 substituents selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido.
• R a and R may be the same or different and and are chosen from hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.
• alkylene refers to a diradical of a branched or unbranched saturated hydrocarbon chain, preferably having from 1 to 40 carbon atoms, preferably 1-10 carbon atoms, more preferably 1 -6 carbon atoms. This term is exemplified by groups such as methylene (-CH -), ethylene (-CH 2 CH 2 -), the propylene isomers (e.g., -CH 2 CH 2 CH 2 - and -CH(CH 3 )CH 2 -) and the like.
• substituted alkylene refers to: (a) an alkylene group as defined above having from 1 to 5 substituents selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino (including, for example, N- glucosaminecarbonyl, benzoylamino, biphenylcarbonylamino, and the like), acyloxy, amino, aminoacyl, aminoacyloxy, oxyacylamino, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, thioaryloxy, heteroaryl, heteroaryloxy, thioheteroaryloxy, heterocyclic, heterocyclooxy
• substituted alkylene groups include those where 2 substituents on the alkylene group are fused to form one or more cycloalkyl, substituted cycloalkyl. cycloalkenyl, substituted cycloalkenyl, aryl, heterocyclic or heteroaryl groups fused to the alkylene group.
• alkylene group as defined above that is interrupted by 1-20 atoms or substituents independently chosen from oxygen, sulfur and NR a -, wherein R a is chosen from hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl. cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic; or
• alkylene group as defined above that has both from 1 to 5 substituents as defined above and is also interrupted by 1 -20 atoms as defined above.
• substituted alkylenes are chloromethylene (-CH(Cl)-), aminoethylene
• lower alkylene refers to the group alkylene as defined above having from 1 -6 carbon atoms.
• alkaryl or "aralkyf'refers to the groups -alkylene-aryl and -substituted alkylene-aryl in which alkylene and aryl are as defined herein. Such alkaryl groups are exemplified by benzyl, phenethyl, and the like.
• alkoxy refers to the groups alkyl-O-, alkenyl-O-, cycloalkyl-O-, cycloalkenyl-O-, and alkynyl-O-, where alkyl. alkenyl, cycloalkyl, cycloalkenyl, and alkynyl are as defined herein.
• Preferred alkoxy groups are alkyl-O- and include, by way of example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy. sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like
• substituted alkoxy refers to the groups substituted alkyl-O-, substituted alkenyl-O-, substituted cycloalkyl-O-, substituted cycloalkenyl-O-, and substituted alkynyl-O- where substituted alkyl, substituted alkenyl, substituted cycloalkyl, substituted cycloalkenyl and substituted alkynyl are as defined herein.
• alkylalkoxy refers to the groups -alkylene-O-alkyl, alkylene-O-substituted alkyl, substituted alkylene-O-alkyl and substituted alkylene-O- substituted alkyl wherein alkyl.
• substituted alkyl, alkylene and substituted alkylene are as defined herein.
• Examples of such groups are methylenemethoxy (-CH 2 OCH ), ethylenemethoxy (-CH 2 CH 2 OCH ), n-propylene-iso-propoxy (-CH 2 CH 2 CH 2 OCH(CH 3 )2), methylene-t-butoxy (-CH 2 -O-C(CH 3 ) 3 ) and the like.
• alkylthioalkoxy refers to the group -alkylene-S-alkyl, alkylene-S-substituted alkyl, substituted alkylene-S-alkyl and substituted alkylene-S- substituted alkyl wherein alkyl, substituted alkyl, alkylene and substituted alkylene are as defined herein.
• Preferred alkylthioalkoxy groups are alkylene-S-alkyl and include, by way of example, methylenethiomethoxy (-CH 2 SCH 3 ), ethylenethiomethoxy
• Alkenyl refers to a monoradical of a branched or unbranched unsaturated hydrocarbon preferably having from 2 to 40 carbon atoms, preferably 2-10 carbon atoms, more preferably 2-6 carbon atoms, and preferably having 1-6 double bonds. This term is further exemplified by such radicals as vinyl, prop-2-enyl, pent-3-enyl. hex-5-enyl. 5- ethyldodec-3,6-dienyl, and the like.
• substituted alkenyl refers to an alkenyl group as defined above having from 1 to 5 substituents selected from the group consisting of alkoxy, substituted alkoxy, acyl, acylamino. acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano.
• halogen hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thiol, thioalkoxy, substituted thioalkoxy, aryl, heteroaryl, heterocyclic, aryloxy, thioaryloxy, heteroaryloxy, thioheteroaryloxy, heterocyclooxy, thioheterocyclooxy, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO 2 -alkyl, -SO 2 -substituted alkyl, -SO 2 -aryl, -SO 2 - heteroaryl, and.
• R a and R b may be the same or different and are chosen from hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.
• Alkenylene refers to a diradical of an unsaturated hydrocarbon, preferably having from 2 to 40 carbon atoms, preferably 2-10 carbon atoms, more preferably 2-6 carbon atoms, and preferably having 1-6 double bonds. This term is further exemplified by such radicals as 1,2-ethenyl, l,3-prop-2-enyl, l,5-pent-3-enyl. 1 ,4-hex-5-enyl, 5-ethyl- l,12-dodec-3,6-dienyl, and the like.
• substituted alkenylene refers to an alkenylene group as defined above having from 1 to 5 substituents. selected from the group consisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyacylamino, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thiol, thioalkoxy, substituted thioalkoxy, aryl. aryloxy, thioaryloxy.
• R a and R b may be the same or different and are chosen from hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl. aryl, heteroaryl and heterocyclic.
• substituted alkenylene groups include those where 2 substituents on the alkenylene group are fused to form one or more cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heterocyclic or heteroaryl groups fused to the alkenylene group.
• Alkynyl refers to a monoradical of an unsaturated hydrocarbon, preferably having from 2 to 40 carbon atoms, preferably 2-10 carbon atoms, more preferably 2-6 carbon atoms, and preferably having 1-6 triple bonds. This term is further exemplified by- such radicals as acetylenyl, prop-2-ynyl, pent-3-ynyl, hex-5-ynyl. 5-ethyldodec-3,6- diynyl, and the like.
• substituted alkynyl refers to an alkynyl group as defined above having from 1 to 5 substituents, selected from the group consisting of alkoxy. substituted alkoxy. acyl, acylamino, acyloxy. amino, aminoacyl, aminoacyloxy.
• oxyacylamino azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, thioaryloxy, heteroaryl, heteroaryloxy, thioheteroaryloxy, heterocyclic, heterocyclooxy, thioheterocycloxy, nitro, -SO-alkyl, -SO-substituted alkyl,
• R a and R b may be the same or different and are chosen from hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.
• Alkynylene refers to a diradical of an unsaturated hydrocarbon radical, preferably having from 2 to 40 carbon atoms, preferably 2- 10 carbon atoms, more preferably 2-6 carbon atoms, and preferably having 1-6 triple bonds. This term is further exemplified by such radicals as l,3-prop-2-ynyl, l,5-pent-3-ynyl, l,4-hex-5-ynyl, 5- ethyl-l,12-dodec-3,6-diynyl, and the like.
• acyl refers to the groups -CHO, alkyl-C(O)-, substituted alkyl-C(O)-. cycloalkyl-C(O)-, substituted cycloalkyl-C(O)-, cycloalkenyl-C(O)-, substituted cycloalkenyl-C(O)-, aryl-C(O)-, heteroaryl-C(O)- and heterocyclic-C(O)- where alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl and heterocyclic are as defined herein.
• acylamino refers to the group -C(O)NRR where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, heterocyclic or where both R groups are joined to form a heterocyclic group (e.g.. morpholino) wherein alkyl. substituted alkyl, aryl, heteroaryl and heterocyclic are as defined herein.
• aminoacyl refers to the group -NRC(O)R where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl and heterocyclic are as defined herein.
• aminoacyloxy refers to the group -NRC(O)OR where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl and heterocyclic are as defined herein.
• acyloxy refers to the groups alkyl-C(O)O-, substituted alkyl-C(O)O-, cycloalkyl-C(O)O-, substituted cycloalkyl-C(O)O-, aryl-C(O)O-, heteroaryl-C(O)O-, and heterocyclic-C(O)O- wherein alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, heteroaryl, and heterocyclic are as defined herein.
• aryl refers to an unsaturated aromatic carbocyclic group of from 6 to 20 carbon atoms having a single ring (e.g., phenyl) or multiple condensed (fused) rings (e.g., naphthyl or anthryl).
• aryl groups can optionally be substituted with from 1 to 5 substituents selected from the group consisting of acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy.
• aryloxy refers to the group aryl-O- wherein the aryl group is as defined above including optionally substituted aryl groups as also defined above.
• arylene refers to a diradical derived from aryl or substituted aryl as defined above, and is exemplified by 1,2-phenylene, 1 ,3-phenylene, 1 ,4-phenylene, 1.2- naphthylene and the like.
• carboxyalkyl refers to the group “-C(O)Oalkyl” where alkyl is as defined above.
• cycloalkyl refers to cyclic alkyl groups of from 3 to 20 carbon atoms having a single cyclic ring or multiple condensed rings.
• Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantanyl, and the like.
• substituted cycloalkyl refers to cycloalkyl groups having from 1 to 5 substituents selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, cycloalkenyl, substituted cycloalkenyl.
• acyl acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO- substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO -alkyl, -SO 2 -substituted alkyl, -SO 2 -aryl, -SO 2 -heteroaryl, and NR a R b , wherein R a
• cycloalkenyl refers to cyclic alkenyl groups of from 4 to 20 carbon atoms having a single cyclic ring or fused rings and at least one point of internal unsaturation.
• suitable cycloalkenyl groups include, for instance, cyclobut-2- enyl, cyclopent-3-enyl, cyclooct-3-enyl and the like.
• substituted cycloalkenyl refers to cycloalkenyl groups having from 1 to 5 substituents selected from the group consisting of alkoxy. substituted alkoxy. cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl.
• cycloalkyl alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.
• halo or “halogen” refers to fluoro, chloro, bromo and iodo.
• Haloalkyl refers to alkyl as defined above substituted by 1 -4 halo groups as defined above, which may be the same or different, such as 3-fluorododecyl, 12,12,12- trifluorododecyl, 2-bromooctyl, -3-bromo-6-chloroheptyl, and the like.
• heteroaryl refers to an aromatic group of from 1 to 15 carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfur within at least one ring (if there is more than one ring). Unless otherwise constrained by the definition for the heteroaryl substituent, such heteroaryl groups can be optionally substituted with 1 to 5 substituents selected from the group consisting of acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkoxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl, amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxyl, carboxylalkyl, cyano, halo, nitro, heteroaryl, heteroaryloxy.
• heterocyclic heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, -SO-alkyl, -SO- substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO 2 -alkyl, -SO -substituted alkyl, -SO 2 -aryl, -SO 2 -heteroaryl, trihalomethyl, mono-and di-alkylamino, mono- and NR a R b , wherein R a and R b may be the same or different and are chosen from hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.
• Preferred heteroaryls nclude pyridyl, pyrrolyl and furyl
• heteroaryloxy refers to the group heteroaryl-O-.
• heteroarylene refers to the diradical group derived from heteroaryl or substituted heteroaryl as defined above, and is exemplified by the groups 2,6-pyridylene, 2,4-pyridiylene, 1,2-quinolinylene, 1,8-quinolinylene, 1 ,4-benzofuranylene, 2,5- pyridinylene, 1,3-morpholinylene, 2,5-indolenyl, and the like.
• heterocycle or “heterocyclic” refers to a monoradical saturated or unsaturated group having a single ring or multiple condensed rings, from 1 to 40 carbon atoms and from 1 to 10 hetero atoms, preferably 1 to 4 heteroatoms, selected from nitrogen, sulfur, phosphorus, and/or oxygen within the ring.
• heterocyclic groups can be optionally substituted with 1 to 5, and preferably 1 to 3 substituents, selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl
• nitrogen heterocycles and heteroaryls include, but are not limited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purin.e, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline.
• cinnoline pteridine, carbazole, carboline, phenanthridine, acridine, phenanthrohne, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, morpholino, piperidinyl, tetrahydrofuranyl, and the like as well as N-alkoxy-nitrogen containing heterocycles.
• a preferred class of heterocyclics include “crown compounds” which refers to a specific class of heterocyclic compounds having one or more repeating units of the formula [-(CH2-)mY-] where m is equal to or greater than 2, and Y at each separate occurrence can be O, N, S or P.
• Examples of crown compounds include, by way of example only, [-(CH 2 ) 3 -NH-] 3 , [-((CH 2 )2-O) 4 -((CH 2 )2-NH)2] and the like. Typically such crown compounds can have from 4 to 10 heteroatoms and 8 to 40 carbon atoms.
• heterocycloalkyl refers to heterocycle as defined above linked to alkyl as defined above, for example piperidin-4-ylmethyl, oxazolidin-2-one-3-ylethyl, and the like.
• heterocyclooxy refers to the group heterocyclic-O-.
• thioheterocyclooxy refers to the group heterocyclic-S-.
• heterocyclene refers to the diradical group derived from a heterocycle as defined herein, and is exemplified by the groups 2,6-morpholino, 2,5-morpholino and the like.
• oxyacylamino refers to the group -OC(O)NRR where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl and heterocyclic are as defined herein.
• thiol refers to the group -SH.
• thioalkoxy refers to the group -S-alkyl.
• substituted thioalkoxy refers to the group -S-substituted alkyl.
• thioaryloxy refers to the group aryl-S- wherein the aryl group is as defined above including optionally substituted aryl groups also defined above.
• heteroaryloxy refers to the group heteroaryl-S- wherein the heteroaryl group is as defined above including optionally substituted aryl groups as also defined above.
• any ofthe above groups which contain one or more substituents it is understood, of course, that such groups do not contain any substitution or substitution patterns which are sterically impractical and/or synthetically non-feasible.
• the compounds of this invention include all stereochemical isomers arising from the substitution of these compounds.
• Alkyl optionally interrupted by 1-5 atoms chosen from O, S, or N refers to alkyl as defined above in which the carbon chain is interrupted by O, S, or N.
• ethers, sulfides, and amines for example 1-methoxydecyl, 1-pentyloxynonane, l-(2- isopropoxyethoxy)-4-methylnonane, l-(2-ethoxyethoxy)dodecyl, 2-(t-butoxy)heptyl, 1- pentylsulfanylnonane, nonylpentylamine, and the like.
• Heteroarylalkyl refers to heteroaryl as defined above linked to alkyl as defined above, for example pyrid-2-ylmethyl. 8-quinolinylpropyl. and the like.
• Optional or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not.
• optionally substituted alkyl means that alkyl may or may not be substituted by those groups enumerated in the definition of substituted alkyl.
• Ligand denotes a compound that is a binding partner for a 5-HT receptor, preferably a 5-HT1 receptor, and is bound thereto by complementarity.
• the specific region or regions ofthe ligand that is (are) recognized by the 5-HT receptor is designated as the "ligand domain".
• a ligand may be either capable of binding to a 5-HT receptor by itself, or may require the presence of one or more non-ligand components for binding (e.g. Ca ⁇ ' Mg ⁇ * . or a water molecule is required for the binding of a ligand domain to various receptors). Examples of ligands useful in this invention, including the triptan class, are given below.
• ligands is not intended to be limited to compounds known to be useful as 5-HT agonists, antagonists, modulators, or the like, (for example, known drugs).
• ligand can equally apply to a molecule that is not normally recognized as having useful properties related to binding to a 5-HT receptor, in that ligands that exhibit minimally useful properties as monomers can be highly active as multibinding agents, due to the biological benefit (increased biological effect) conferred by multivalency.
• the primary requirement for a ligand as defined herein is that it has a ligand domain as defined above.
• Preferred ligands include AH-25086, Almotriptan (LAS-31416), Alniditan. ALX- 0646, Avitriptan (BMS- 180048), CP 122.288, 5-CT, Eletriptan (UK- 1 16044), IS- 159. LY-334370, L-694,247, ⁇ -Methyl-5-HT, 2-Methyl-5-HT, Naratriptan, Oxymetazoline, PNU-109291, PNU-142633, Rizatriptan (MK-462), (L-741 ,519), S-9977, S-20749, SB209509 (VML251), Sumatriptan, VML-251, Zolmitriptan.
• Alnespirone (S-20499), B- 20991 , BAY x 3702. Buspirone, Cisapride, CGS-12066B, CP93129, Gepirone, GR 127935, Ipsaperone, MDL 74,721, 5-Methoxytryptamine, Methysergide, Metoclopramide, MKC-242, Mosapride citrate, 8-OH-DPAT. Quipazine. Renzapride, RS67506, Tandospirone. VB20B7, Zelmac, (R) Zacopride.Zacopride, Zalospirone, Umespirone, Enilospirone, WY-48723, SUN-8399. Flesinoxan.
• Formula I is intended to include racemic ligands L and racemic linker X, as well as the individual stereoisomers ofthe ligands and linkers, including enantiomers and non-racemic mixtures thereof.
• the scope ofthe invention as described and claimed encompasses the racemic forms ofthe ligands and linkers as well as the individual enantiomers and non-racemic mixtures thereof.
• Multibinding agent or “multibinding compound” as used herein refers to a compound that is capable of multivalency as defined herein, and which has 2-10 ligands as defined herein, which may be the same or different, connected by one or more covalent linker or linkers, which may be the same or different, preferably from 1 -20 in number.
• a multibinding agent provides an improved biological a id/or therapeutic effect as measured against that achieved by the same number of unlinked ligands available for binding to the ligand binding site ofthe 5-HT receptor.
• Examples of increased biological and/or therapeutic effect with respect to the target include, for example, increased specificity, increased affinity, increased selectivity, increased potency, increased efficacy, increased therapeutic index, a change in the duration of action, decreased toxicity, decreased side effects, improved bioavailability, improved pharmacokinetics. improved activity spectrum, and the like.
• the multibinding compounds of the invention exhibit one or more ofthe foregoing effects.
• “Potency” as used herein refers to the minimum concentration at which a ligand is able to achieve a desirable biological or therapeutic effect.
• the potency of a ligand is typically proportional to its affinity for its ligand binding site. In some cases, the potency may be non-linearly correlated with its affinity.
• the dose-response curve of each is determined under identical test conditions (e.g., in an in vitro or in vivo assay or in an appropriate animal model. The finding that the multibinding agent produces an equivalent biological or therapeutic effect at a lower concentration than the aggregate unlinked ligand (e.g., on a per weight, per mole, or per ligand basis) is indicative of enhanced potency.
• Univalency refers to a single binding interaction between the ligand domain of one ligand as defined herein with the ligand recognition site of a 5-HT receptor. It should be noted that a compound having multiple copies of a ligand (or ligands) exhibits univalency when only one ligand of that compound is interacting with a ligand binding site. Examples of univalent interactions are depicted below.
• Multivalency refers to the concurrent binding of 2 to 10 linked ligands (which may be the same or different) and two or more corresponding ligand binding sites of one or more 5-HT receptors. Accordingly, two ligands connected by a linker that bind concurrently to two ligand binding sites of one or more 5-HT receptors would be considered to be a bivalent compound; similarly, three ligands thus connected provide a trivalent compound.
• An example of trivalent binding, illustrating a multibinding agent bearing three ligands is shown below. It should be understood that all compounds that contain multiple copies of a ligand attached to a linker (or linkers) do not necessarily exhibit the phenomena of multivalency, i.e.
• multibinding agent that improved biological and/or therapeutic effect of the multibinding agent is obtained as measured against that produced by the same number of unlinked ligands available for binding to a ligand binding site.
• the ligand domains ofthe ligands that are connected by a linker have to be presented to their appropriate receptor(s) (i.e. the ligand binding sites) by the linker in a specific manner in order to bring about the desired ligand-orienting result, and thus produce a multibinding event.
• the term "multimeric ligand compound” refers to multiple copies of a ligand attached to a linker (or linkers) that may or may not exhibit the phenomena of multivalency.
• Multimeric ligand compound library refers to the collection of multimeric ligand compounds that are provided by the synthetic methods disclosed herein.
• Selectivity or “specificity” is a measure ofthe binding preferences of a ligand for different receptors and/or different ligands for the same receptor.
• the selectivity of a ligand with respect to its target receptor relative to another receptor is given by the ratio ofthe respective values of Kd (i.e., the dissociation constants for each ligand-receptor complex), or in cases where a biological effect is observed below the K d , selectivity is given by the ratio ofthe respective EC 50 s (i.e. the concentrations that produce 50% ofthe maximum response for the ligand interacting with the two distinct receptors).
• ligand recognition site or "ligand binding site” as used herein denotes the site on a 5-HT receptor that recognizes a ligand domain and provides a binding partner for a ligand.
• the ligand binding site may be defined by monomeric or multimeric structures. This interaction may be capable of producing a unique biological effect, for example, agonism, antagonism, modulatory effect, and the like, or may maintain an ongoing biological effect.
• ligand binding sites of receptors that participate in biological multivalent binding interactions are constrained to varying degrees by their intra- and intermolecular associations (e.g., they may be covalently joined in a single or multiple structure, noncovalently associated in a multimeric structure, embedded in a membrane or polymeric matrix, and so on) and therefore have less relative translational and rotational freedom than if the same receptors were present as monomers in solution.
• the terms "agonism” and “antagonism” are well known in the art.
• modulatory effect we mean the ability of a ligand to change the biological effect of an agonist or antagonist through binding to a receptor.
• inert organic solvent or “inert solvent” mean a solvent inert under the conditions ofthe reaction being described in conjunction therewith [including, for example, benzene, toluene, acetonitrile. tetrahydrofuran (“THF”), dimethylformamide (“DMF”), chloroform (“CHC1 3 "), methylene chloride (or dichloromethane or "CH 2 CI2), diethyl ether, ethyl acetate, acetone, methylethyl ketone, methanol, ethanol, propanol, isopropanol, tert-butanol, dioxane, pyridine, and the like].
• THF tetrahydrofuran
• DMF dimethylformamide
• CHC1 3 chloroform
• CH 2 CI2 methylene chloride
• diethyl ether diethyl ether, ethyl acetate, acetone, methylethyl
• the solvents used in the reactions ofthe present invention are inert solvents.
• “Pharmaceutically acceptable salt” means those salts which retain the biological effectiveness and properties ofthe multivalent compounds ofthe invention, and which are not biologically or otherwise undesirable.
• the multivalent compounds ofthe invention are capable of forming both acid and base salts by virtue ofthe presence of amino and carboxyl groups respectively.
• Pharmaceutically acceptable base addition salts may be prepared from inorganic and organic bases.
• Salts derived from inorganic bases include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, and magnesium salts.
• 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, and cyclic amines, including isopropylamine, trimethyl amine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine, purines, piperazine, piperidine, and N- ethylpiperidine.
• carboxylic acid derivatives would be useful in the practice of this invention, for example carboxylic acid amides, including carboxamides, lower alkyl carboxamides, di(lower alkyl) carboxamides. and the like.
• Salts derived from inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like.
• Salts derived from organic acids include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic 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.
• treatment covers any treatment of a condition or disease in an animal, particularly a mammal, more particularly a human, and includes: (i) preventing the disease or condition from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it; (ii) inhibiting the disease or condition, i.e. arresting its development; (iii) relieving the disease or condition, i.e. causing regression ofthe condition; or. (iv) relieving the conditions caused by the disease, i.e. symptoms ofthe disease.
• disease or condition which is alleviated by treatment with a multibinding agent covers all conditions and disease states which are generally acknowledged in the art to be usefully treated with the ligands as defined in general, and those disease states which have been found to be usefully treated by the specific multibinding agents of our invention, including the compounds of Formula I.
• the term covers prophylactic treatment as well as relief or regression ofthe disease. It also covers the treatment of conditions that are not necessarily considered as disease states, for example the use of multibinding agents as contraceptives, as pregnancy limiting agents, for the treatment of insomnia, treatment of obesity, and the like.
• Such disease states include, but are not limited to, treatment of a mammal for modifying physiological functions related to sleep, appetite, pain, movement, and temperature regulation, and includes disease states such as migraine, headache, itch, motion sickness, depression, emesis, memory loss, anxiolytic disorders, obesity, gastrointestinal disorders, irritable bowel syndrome, and the like.
• the term "therapeutically effective amount” refers to that amount of a multibinding agent, for example a compound of Formula I, that is sufficient to effect treatment, as defined above, when administered to a mammal or avian in need of such treatment.
• the therapeutically effective amount will vary depending on the subject and disease state being treated, the severity of the affliction and the manner of administration. and the like, and may be determined routinely by one of ordinary skill in the art.
• protecting group refers to any group which when bound to one or more hydroxyl, thiol, amino or carboxyl groups ofthe compounds prevents reactions from occurring at these groups and which protecting group can be removed by conventional chemical or enzymatic steps to reestablish the hydroxyl. thio, amino or carboxyl group.
• the particular removable blocking group employed is not critical and preferred removable hydroxyl blocking groups include conventional substituents such as allyl, benzyl, acetyl, chloroacetyl.
• Preferred removable amino blocking groups include conventional substituents such as t-butyoxycarbonyl (t-BOC), benzyloxycarbonyl (CBZ), fluorenylmethoxycarbonyl (FMOC), allyloxycarbonyl (ALOC) and the like, which can be removed by conventional conditions compatible with the nature ofthe product.
• t-BOC t-butyoxycarbonyl
• CBZ benzyloxycarbonyl
• FMOC fluorenylmethoxycarbonyl
• ALOC allyloxycarbonyl
• Preferred carboxyl protecting groups include esters such as methyl, ethyl, propyl, t-butyl etc. which can be removed by mild conditions compatible with the nature of the product.
• Esters such as methyl, ethyl, propyl, t-butyl etc. which can be removed by mild conditions compatible with the nature of the product.
• Linker or “linkers” as used herein, identified where appropriate by the symbol
• X refers to a group or groups that covalently link(s) from 2-10 ligands (as defined herein) in a manner that provides a compound capable of multivalency.
• the linker is a ligand domain orienting entity that permits attachment of multiple copies of ligands (which may be the same or different) thereto. The extent to which multivalent binding is realized depends upon the efficiency with which the linker that joins the ligands permits the ligand domains to be presented to the ligand recognition sites substrates). Beyond presenting ligand domains for multivalent interactions with receptors, the linker spatially constrains these interactions to occur within dimensions defined by the linker.
• linker valency, geometry, orienting capabilities, size, flexibility, chemical composition
• linker does not include solid inert supports such as beads, resins, glass particles, rods, fibers, and the like, but it should be understood that the multibinding compounds of the invention can be attached to a solid support if desired to provide, for example, a material useful for separation and purification processes (e.g. affinity chromatography).
• the ligands are covalently attached to the linker or linkers using conventional chemical techniques, for example reaction between a carboxylic acid and an amine to form an amide, an amine and a sulfonyl halide to form a sulfonamide. an alcohol or phenol with an alkyl or aryl halide to form an ether, and the like.
• the linker (or linkers) is attached to the ligand at a position such that the ligand domain is permitted to orient itself appropriately in order to bind to the ligand binding site.
• the linker is attached to an existing sidechain of a ligand.
• the linker X is normally defined as including the sidechain, and the ligand is defined as excluding the sidechain.
• the compound of Formula I having a structure:
• R 3 is the linking position; and the linker is:
• the relative orientation in which the ligand domains are displayed derives from the particular point or points of attachment ofthe ligands to the linker, and on the framework geometry.
• the determination of where acceptable substitutions can be made on a ligand is typically based on prior knowledge of structure-activity relationships ofthe ligand and/or congeners and/or structural information about ligand-receptor complexes (e.g., from X-ray crystallography, NMR). Such positions and the synthetic methods for covalent attachment are well known in the art. Suitable linkers are discussed below.
• the multibinding agent is a bivalent compound, in which two ligands are covalently linked.
• Bio effect includes, but is not limited to, increased affinity, increased selectivity, increased potency, increased efficacy, increased duration of action, decreased toxicity, and the like.
• the methods described above lend themselves to combinatorial approaches for selecting compounds that have multibinding properties related to the 5-HT receptor, in particular the 5HT1 receptor, from a library of multimeric compounds.
• factors such as the proper juxtaposition of the individual 5HT ligands of a multibinding compound with respect to the relevant array of binding sites on a target or targets is important in optimizing the interaction of the multibinding compound with its target(s) and to maximize the biological advantage through multivalency.
• One approach is to identify a library of candidate multibinding compounds with properties spanning the multibinding parameters that are relevant for a particular target. These parameters include: (1) the identity of ligand(s), (2) the orientation of ligands, (3) the valency ofthe construct, (4) linker length, (5) linker geometry, (6) linker physical properties, and (7) linker chemical functional groups. Libraries of multimeric compounds potentially possessing multibinding properties
• a single 5-HT ligand or set of 5-HT ligands is (are) selected for inco ⁇ oration into the libraries of candidate multibinding compounds.
• the only requirement for the ligands chosen is that they are capable of interacting with a 5-HT receptor(s). preferably a 5-HTl receptor.
• 5-HTl ligands may be known drugs, modified forms of known drugs, substructures of known drugs or substrates of modified forms of known drugs (which are competent to interact with the target), or other compounds.
• 5-HTl ligands are preferably chosen based on known favorable properties that may be projected to be carried over to or amplified in multibinding forms.
• 5-HT ligands which display an unfavorable property from among the previous list may obtain a more favorable property through the process of multibinding compound formation; i.e., 5-HT ligands should not necessarily be excluded on such a basis.
• a 5-HT ligand that is not sufficiently potent at a particular target so as to be efficacious in a human patient may become highly potent and efficacious when presented in multibinding form.
• a 5-HT ligand that is potent and efficacious but not of utility because of a non-mechanism-related toxic side effect may have increased therapeutic index (increased potency relative to toxicity) as a multibinding compound.
• Compounds that exhibit short in vivo half-lives may have extended half-lives as multibinding compounds.
• Physical properties of 5-HT ligands that limit their usefulness e.g. poor bioavailability due to low solubility, hydrophobicity. hydrophilicity
• each 5-HT ligand at which to attach the 5-HT ligand to the linker.
• the selected points on the 5-HT ligand/linker for attachment are functionalized to contain complementary reactive functional groups. This permits probing the effects of presenting the 5-HT ligands to their receptor(s) in multiple relative orientations, an important multibinding design parameter.
• the only requirement for choosing attachment points is that attaching to at least one of these points does not abrogate activity ofthe 5-HT ligand.
• Such points for attachment can be identified by structural information when available. For example, inspection of a co-crystal structure of a protease inhibitor bound to its target allows one to identify one or more sites where linker attachment will not preclude the enzyme inhibitor interaction.
• positions of attachment that do abrogate the activity ofthe monomeric 5-HT ligand may also be advantageously included in candidate multibinding compounds in the library provided that such compounds bear at least one 5- HT ligand attached in a manner which does not abrogate intrinsic activity. This selection derives from, for example, heterobivalent interactions within the context of a single target molecule.
• the most favorable orientation for interaction of the second 5-HTl ligand molecule with the receptor/matrix may be achieved by attaching it to the linker at a position which abrogates activity of the 5-HTl ligand at the formal agonist binding site.
• Another way to consider this is that the SAR of individual 5-HTl ligands within the context of a multibinding structure is often different from the SAR of those same 5-HTl ligands in momomeric form.
• the most preferred types of chemical linkages are those that are compatible with the overall structure ofthe 5-HT ligand (or protected forms ofthe 5-HT ligand) readily and generally formed, stable and intrinsically inocuous under typical chemical and physiological conditions, and compatible with a large number of available linkers.
• Amide bonds, ethers, amines, carbamates, ureas, and sulfonamides are but a few examples of preferred linkages.
• Linkers spanning relevant multibinding parameters through selection of valency, linker length, linker geometry, rigidity, physical properties, and chemical functional groups
• the selection of linkers employed in this library of linkers takes into consideration the following factors:
• linkers In most instances the library of linkers is initiated with divalent linkers.
• divalent linkers or constructs are also typically of modest size such that they retain the desirable biodistribution properties of small molecules.
• Linker length Linkers are chosen in a range of lengths to allow the spanning of a range of inter-ligand distances that encompass the distance preferable for a given divalent interaction. In some instances the preferred distance can be estimated rather precisely from high-resolution structural information of targets, typically enzymes and soluble receptor targets.
• preferred linker distances are 2-20 angstroms, with more preferred linker distances of 3-12 angstroms. In situations where two binding sites reside on separate (e.g., protein) target sites, preferred linker distances are 20-100 angstroms, with more preferred distances of 30-70 angstroms. Linker geometry and rigidity.
• linker geometry and rigidity are nominally determined by chemical composition and bonding pattern, which may be controlled and are systematically varied as another spanning function in a multibinding array. For example, linker geometry is varied by attaching two 5-HT ligands to the ortho, meta, and para positions of a benzene ring, or in cis- or trans-arrangements at the 1,1- vs. 1,2- vs. 1,3- vs.
• Linker rigidity is varied by controlling the number and relative energies of different conformational states possible for the linker.
• a divalent compound bearing two 5-HT ligands joined by 1,8-octyl linker has many more degrees of freedom, and is therefore less rigid than a compound in which the two 5-HT ligands are attached to the 4,4' positions of a biphenyl linker.
• Linker physical properties The physical properties of linkers are nominally determined by the chemical constitution and bonding patterns ofthe linker, and linker physical properties impact the overall physical properties of the candidate multibinding compounds in which they are included.
• a range of linker compositions is typically- selected to provide a range of physical properties (hydrophobicity, hydrophilicity, amphiphilicity, polarization, acidity, and basicity) in the candidate multibinding compounds.
• the particular choice of linker physical properties is made within the context ofthe physical properties of the 5-HT ligands they join and preferably the goal is to generate molecules with favorable PK/ADME properties.
• linkers can be selected to avoid those that are too hydrophilic or too hydrophobic to be readily absorbed and/or distributed in vivo.
• Linker chemical functional groups are selected to be compatible with the chemistry chosen to connect linkers to the 5-HT ligands and to impart the range of physical properties sufficient to span initial examination of this parameter.
• n being determined by the sum ofthe number of different attachment points for each 5-HT ligand chosen
• m linkers by the process outlined above
• a library of (n!)m candidate divalent multibinding compounds is prepared which spans the relevant multibinding design parameters for a particular target. For example, an array generated from two 5-HT ligands, one which has two attachment points (Al, A2) and one which has three attachment points (Bl, B2, B3) joined in all possible combinations provide for at least 15 possible combinations of multibinding compounds:
• the combinatorial library can employ solid phase chemistries well known in the art wherein the 5-HT ligand and/or linker is attached to a solid support.
• the combinatorial libary is prepared in the solution phase.
• candidate multibinding compounds are optionally purified before assaying for activity by, for example, chromatographic methods (e.g., HPLC).
• Various methods are used to characterize the properties and activities ofthe candidate multibinding compounds in the library to determine which compounds possess multibinding properties. Physical constants such as solubility under various solvent conditions and logD/clogD values can be determined. A combination of NMR spectroscopy and computational methods is used to determine low-energy conformations ofthe candidate multibinding compounds in fluid media. The ability ofthe members of the library to bind to the desired target and other targets is determined by various standard methods, which include radioligand displacement assays for receptor and ion channel targets, and kinetic inhibition analysis for many enzyme targets. In vitro efficacy, such as for receptor agonists and antagonists, ion channel blockers, and antimicrobial activity, can also be determined. Pharmacological data, including oral absorption, everted gut penetration, other pharmacokinetic parameters and efficacy data can be determined in appropriate models. In this way, key structure-activity relationships are obtained for multibinding design parameters which are then used to direct future work.
• the members ofthe library which exhibit multibinding properties can be readily determined by conventional methods. First those members which exhibit multibinding properties are identified by conventional methods as described above including conventional assays (both in vitro and in vivo). Second, ascertaining the structure of those compounds which exhibit multibinding properties can be accomplished via art recognized procedures. For example, each member ofthe library can be encrypted or tagged with appropriate information allowing determination of the structure of relevant members at a later time. See, for example, Dower, et al., International Patent Application Publication No. WO 93/06121; Brenner, et al., Proc. Natl. Acad. Sci., USA, 89:5181 ( 1992); Gallop, et al., U.S. Patent No.
• the structure of relevant multivalent compounds can also be determined from soluble and untagged libaries of candidate multivalent compounds by methods known in the art such as those described by Hindsgaul, et al., Canadian Patent Application No. 2,240,325 which was published on July 1 1 , 1998. Such methods couple frontal affinity chromatography with mass spectroscopy to determine both the structure and relative binding affinities of candidate multibinding compounds to receptors.
• an optional component ofthe process is to ascertain one or more promising multibinding "lead” compounds as defined by particular relative 5-HT ligand orientations, linker lengths, linker geometries, etc. Additional libraries can then be generated around these leads to provide for further information regarding structure to activity relationships. These arrays typically bear more focused variations in linker structure in an effort to further optimize target affinity and/or activity at the target (antagonism, partial agonism, etc.), and or alter physical properties.
• suitable divalent linkers include, by way of example only, those derived from dicarboxylic acids, disulfonylhalides, dialdehydes, diketones, dihalides, diisocyanates,diamines, diols, mixtures of carboxylic acids, sulfonylhalides, aldehydes, ketones, halides, isocyanates, amines and diols.
• carboxylic acid, sulfonylhahde, aldehyde, ketone. halide, isocyanate, amine and diol functional group is reacted with a complementary functionality on the 5-HT ligand to form a covalent linkage.
• complementary functionality is well known in the art as illustrated in the following table:
• First Reactive Group Second Reactive Group Linkage hydroxyl isocyanate urethane amine epoxide ⁇ -hydroxyamine sulfonyl halide amine sulfonamide carboxyl acid amine amide hydroxyl alkyl/aryl halide ether aldehyde amine/NaCNBH 2 amine ketone amine/NaCNBH 2 amine amine isocyanate urea
• Exemplary linkers include those described in the Appendix.
• the compounds ofthe invention are usually administered in the form of pharmaceutical compositions. These compounds can be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular, and intranasal. These compounds are effective as both injectable and oral compositions. Such compositions are prepared in a manner well known in the pharmaceutical art and comprise at least one active compound.
• compositions that contain, as the active ingredient, one or more of the compounds of formula I above associated with one or more pharmaceutically acceptable carriers.
• the active ingredient is usually mixed with an excipient, diluted by an excipient or enclosed within such a carrier which can be in the form of a capsule, sachet, paper or other container.
• the excipient serves as a diluent, it can be a solid, semi- solid, or liquid material, which acts as a vehicle, carrier or medium for the active ingredient.
• compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.
• the particle size is normally adjusted by milling to provide a substantially uniform distribution in the formulation, e.g. about 40 mesh.
• suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methyl cellulose.
• the formulations can additionally include: lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as methyl- and propylhydroxy-benzoates; sweetening agents; and flavoring agents.
• lubricating agents such as talc, magnesium stearate, and mineral oil
• wetting agents such as talc, magnesium stearate, and mineral oil
• emulsifying and suspending agents preserving agents such as methyl- and propylhydroxy-benzoates
• sweetening agents and flavoring agents.
• the compositions ofthe invention can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient by employing procedures known in the art.
• the compositions are preferably formulated in a unit dosag e form, each dosage containing from about 0.1 mg to about 1 g, more usually about 1 to about 100 mg, ofthe active ingredient.
• unit dosage forms refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient.
• the compound of formula I above is employed at no more than about 20 weight percent of the pharmaceutical composition, more preferably no more than about 15 weight percent, with the balance being pharmaceutically inert carrier(s).
• the active compound is effective over a wide dosage range and is generally administered in a pharmaceutically effective amount. It, will be understood, however, that the amount of the compound actually administered will be determined by a physician, in the light ofthe relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the age, weight, and response of the individual patient, the severity ofthe patient's symptoms, and the like.
• the principal active ingredient is mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound of the present invention.
• a solid preformulation composition containing a homogeneous mixture of a compound of the present invention.
• the active ingredient is dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules.
• This solid preformulation is then subdivided into unit dosage forms ofthe type described above containing from, for example, 0.1 to about 500 mg ofthe active ingredient ofthe present invention.
• the tablets or pills ofthe present invention may be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action.
• the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former.
• the two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release.
• enteric layers or coatings such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.
• liquid forms in which the novel compositions of the present invention may be incorporated for administration orally or by injection include aqueous solutions, suitably flavored syrups, aqueous or oil suspensions, and flavored emulsions with edible oils such as corn oil, cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.
• compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders.
• the liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described supra.
• the compositions are administered by the oral or nasal respiratory route for local or systemic effect.
• Compositions in preferably pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizing device or the nebulizing device may be attached to a face mask tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered, preferably orally or nasally, from devices that deliver the formulation in an appropriate manner.
• composition or formulation to be administered will, in any event, contain a quantity ofthe active compound(s) in an amount effective to alleviate the symptoms ofthe subject being treated.
• the multibinding agents of the invention are useful in medical treatments related to the modulation of 5-HT receptors, and accordingly exhibit biological effects well known to those skilled in the art.
• Examples of such activity include the modification of physiological functions related to sleep, appetite, pain, movement, and temperature regulation, and includes disease states such as migraine, headache, itch, motion sickness, depression, emesis, memory loss, anxiolytic disorders, obesity, gastrointestinal disorders, irritable bowel syndrome, and the like.
• the multibinding agents of the invention are useful in medical treatments and exhibit biological effects that can be demonstrated in routine tests well known to those skilled in the art. For example, measurement of in vitro binding assay/affinity using radioligand binding assays, evaluation of agonist partial agonist character in in vitro functional assays, measurement of functional activity in in vitro reporter gene assay, measurement of efficacy ex vivo functional assays, and utilization of in vivo models. See Example 28.
• the linker when covalently attached to multiple copies of the 5-HT ligands, provides a biocompatible, substantially non-immunogenic multibinding agent.
• the biological effects of the multibinding agent is highly sensitive to the valency, geometry, composition, size, flexibility or rigidity, the presence or absence of anionic or cationic charge, and similar considerations (including hydrophilicity and hydrophobicity as discussed below) with respect to the linker. Accordingly, the linker is preferably chosen to maximize the desired biological effect.
• the linker may be biologically "neutral", i.e. not itself contribute any biological activity to the compound of Formula I, or it may be chosen to enhance the biological effect ofthe molecule.
• the linker may be chosen from any organic molecule that orients two or more ligands to the 5-HT receptors, and permits multivalency.
• the linker can be considered as a "framework" on which the ligands are arranged in order to bring about the desired ligand-orienting result, and thus produce a multibinding agent.
• different orientations can be achieved by including in the framework groups containing monocyclic or polycyclic groups, including aryl and heteroaryl groups, or structures inco ⁇ orating one or more carbon-carbon multiple bonds (i.e., alkenes and alkynes).
• Other groups can also include oligomers and polymers which are branched- or straight-chain species.
• rigidity is imparted by the presence of cyclic groups (e.g., aryl, heteroaryl, cycloalkyl, heterocycles. etc.).
• the ring is a six-or ten membered ring.
• the ring is an aromatic group such as, for example, phenyl or naphthyl.
• frameworks can be designed to provide preferred orientations of the ligands.
• Such frameworks may be represented by using an array of dots (as shown below) wherein each dot may potentially be an atom, such as C, O, N. S. P, H, F, Cl, Br, and F, or the dot may alternatively indicate the absence of an atom at that position.
• the framework is illustrated as a two dimensional array in the following diagram, although clearly the framework is a three dimensional array in practice:
• Each dot is either an atom, chosen from carbon, hydrogen, oxygen, nitrogen, sulfur, phosphorus, or halogen, or the dot represents a point in space (i.e. an absence of an atom). Only certain atoms on the grid have the ability to act as an attachment point for the ligands, namely C, O, N, S, and P.
• Atoms can be connected to each other via bonds (single, double, or triple with acceptable resonance and tautomeric forms), with regard to the usual constraints of chemical bonding.
• Ligands may be attached to the framework via single, double, or triple bonds (with chemically acceptable tautomeric and resonance forms).
• Multiple ligand groups (2 to 10) can be attached to the framework such that the minimal, shortest path distance between adjacent ligand groups does not exceed 100 atoms or 40 angstroms.
• Nodes (1,2), (2,0), (4,4), (5,2), (4,0), (6,2), (7,4). (9.4). (10,2), (9.0), (7,0) all represent carbon atoms.
• Node (10,0) is a chlorine atom. All other nodes (or dots) are points in space (i.e. represent an absence of atoms).
• Hydrogen atoms are affixed to nodes (2.4), (4,4), (4,0). (2,0), (7,4), (10,2), and (7,0).
• the carbon atoms present are connected by either single or double bonds, taking into consideration the principle of resonance and or tautomerism.
• Display vectors around similar central core structures such as phenyldiacetylene and cyclohexane dicarboxylic acid can be varied, as can the spacing ofthe ligand domain from the core structure (i.e., the length of the attaching moiety). It is to be noted that core structures other than those shown here can be used for determining the optimal framework display orientation ofthe ligands. The process may require the use of multiple copies ofthe same central core structure or combinations of different types of display cores.
• the physical properties of the linker can be optimized by varying the chemical composition.
• the composition of a linker can be varied in numerous ways to achieve the desired physical properties. It can therefore be seen that there is a plethora of possibilities for the composition of a linker.
• linkers include aliphatic moieties, aromatic moieties, steroidal moieties, peptides, and the like. Specific examples are peptides or polyamides. hydrocarbons, aromatic groups, ethers, lipids, cationic or anionic groups, or a combination thereof, and many specific examples of linkers are shown in the Appendix.
• linker can be modified by the addition or insertion of ancillary groups into the linker, for example, to change solubility ofthe multibinding agent (in water, fats, lipids. biological fluids, etc.). hydrophobicity, hydrophilicity, linker flexibility, antigenicity, molecular size, molecular weight, in vivo half-life, in vivo distribution, biocompatability, immunogenicity, stability, and the like.
• the introduction of one or more poly or preferably oIigo(ethylene glycol) (PEG) groups onto the linker enhances hydrophilicity and water solubility ofthe multibinding agent, increases both molecular weight and molecular size and, depending on the nature ofthe unPEGylated linker, may increase the in vivo retention time. Further, PEG may decrease antigenicity and potentially enhances the overall rigidity ofthe linker. Ancillary groups that enhance the water solubility/hydrophilicity of the linker are useful in practicing the present invention.
• ancillary groups such as, for example, poly(ethylene glycol), alcohols, polyols (e.g., glycerin, glycerol propoxylate, saccharides, including mono-, oligo- and polysaccharides, etc.), carboxylates, polycarboxylates (e.g., polyglutamic acid, polyacrylic acid, etc.), amines, polyamines (e.g., polylysine, poly(ethyleneimine), etc) to enhance the water solubility and/or hydrophilicity ofthe compounds of Formula I.
• the ancillary group used to improve water solubility/hydrophilicity will be a polyether.
• the ancillary group will be a poly(ethylene glycol).
• lipophilic ancillary groups within the structure of the linker to enhance the lipophilicity and/or hydrophobicity of the compounds of Formula I is within the scope ofthe present invention.
• Lipophilic groups of use in practicing the instant invention include, but are not limited to, aryl and heteroaryl groups.
• the aromatic groups may be either unsubstituted or substituted with other groups, but are at least substituted with a group which allows their covalent attachment to the linker.
• Other lipophilic groups of use in practicing the instant invention include fatty acid derivatives which do not form bilayers in aqueous medium until higher concentrations are reached.
• lipid refers to any fatty acid derivative that is capable of forming a bilayer or micelle such that a hydrophobic portion ofthe lipid material orients toward the bilayer while a hydrophilic portion orients toward the aqueous phase. Hydrophilic characteristics derive from the presence of phosphato, carboxylic, sulfato, amino, sulfhydryl, nitro, and other like groups.
• Hydrophobicity could be conferred by the inclusion of groups that include, but are not limited to, long chain saturated and unsaturated aliphatic hydrocarbon groups of up to 20 carbon atoms and such groups substituted by one or more aryl, heteroaryl, cycloalkyl and/or heterocyclic group(s).
• Preferred lipids are phosphoglycerides and sphingolipids, representative examples of which include phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoyl phosphatidylcholine, lysophosphatidylcholine, lysophosphatidyl-ethanolamine, dipalmitoylphosphatidylcholine, dioleoylphosphatidylcholine, distearoyl- phosphatidylcholine or dilinoleoylphosphatidylcholine could be used.
• lipid Other compounds lacking phosphorus, such as sphingolipid and glycosphingolipid families are also within the group designated as lipid. Additionally, the amphipathic lipids described above may be mixed with other lipids including triglycerides and sterols.
• the flexibility ofthe linker can be reduced by the inclusion of ancillary groups which are bulky and/or rigid.
• the presence of bulky or rigid groups can hinder free rotation about bonds in the linker or bonds between the linker and the ancillary group(s) or bonds between the linker and the functional groups.
• Rigid groups can include, for example, those groups whose conformational lability is restrained by the presence of rings and/or multiple bonds, for example, aryl, heteroaryl, cycloalkyl, and/or heterocyclic.
• Other groups which can impart rigidity include polymeric groups such as oligo- or polyproline chains. Rigidity can also be imparted electrostatically.
• the similarly charged ancillary groups will force the presenter linker into a configuration affording the maximum distance between each of the like charges.
• the energetic cost of bringing the like-charged groups closer to each other will tend to hold the linker in a configuration that maintains the separation between the like-charged ancillary groups.
• ancillary groups bearing opposite charges will tend to be attracted to their oppositely charged counte ⁇ arts and potentially may enter into both inter- and intramolecular ionic bonds. This non-covalent bonding mechanism will tend to hold the linker into a conformation which allows bonding between the oppositely charged groups.
• Rigidity may also be imparted by internal hydrogen bonding, or by hydrophobic collapse.
• Bulky groups can include, for example, large atoms and/or ions (e.g., iodine, sulfur, metal ions, etc.) groups containing large atoms, polycyclic groups, including aromatic groups, non-aromatic groups and structures inco ⁇ orating one or more carbon- carbon multiple bonds (i.e., alkenes and alkynes).
• Bulky groups can also include oligomers and polymers which are branched- or straight-chain species. Species that are branched are expected to increase the rigidity ofthe structure more per unit molecular weight gain than are straight-chain species.
• rigidity is imparted by the presence of cyclic groups (e.g., aryl, heteroaryl, cycloalkyl, heterocycles, etc.).
• cyclic groups e.g., aryl, heteroaryl, cycloalkyl, heterocycles, etc.
• the ring comprises one or more six-membered rings.
• the ring is an aryl group such as, for example, phenyl or naphthyl.
• the antigenicity of a compound of Formula I may be reduced or eliminated by the use of groups such as, for example, poly(ethylene glycol).
• the multibinding agents of the invention comprise 2-10 ligands attached to a linker that connects the ligands in such a manner that they are presented to the 5-HT receptors for multivalent interactions with the appropriate receptors (ligand binding site). The linker spatially constrains these interactions to occur within dimensions defined by the linker, thus increasing the biological effect ofthe multibinding agent as compared to the same number of individual units of the ligand.
• the multivalent compounds ofthe invention are represented by the empirical formula (L) p (X) q .
• L the empirical formula
• X the ligands can be linked together in order to achieve the objective of multivalency, and a more detailed explanation is given below.
• the linker can be considered as a framework, and it should be understood that the ligands can be attached to this framework at any intermediate point on the framework. and/or on the termini ofthe framework.
• the linker is a linear chain
• a bivalent compound can be constructed by attaching two ligands at the two ends ofthe linear chain, or alternatively attaching two ligands at some intermediate atom along the chain.
• the same considerations apply to the compounds of the present invention containing more than 2 ligands.
• the simplest (and preferred) multibinding agent is a bivalent compound, which can be represented as L-X-L, where L is a ligand and is the same or different, and X is the linker.
• the linker X can be linear or cyclic, or a combination of both linear and cyclic constructs, and that the two ligands may be located at the termini of the linker or may be attached at some intermediate attachment point.
• This concept is diagrammed in the Appendix. The same is true for a trivalent compound, which can also be represented in a linear fashion, i.e.
• L is a ligand and is the same or different at each occurrence, as can X, or a compound comprising three ligands attached to a central core, and thus represented as (L) 3 X, where the linker X could include, for example, an aryl or cycloalkyl group. See the Appendix for a pictorial representation of this concept, in which the shaded objects represent a ligand and the remaining structure represents the linker.
• L i.e. a branched construct analogous to the isomers of butane (n-butyl, sec-butyl, tert- butyl). Alternatively, it could be represented as an aryl or cycloalkyl derivative as above with four ligands attached to the core linker.
• the same principles apply to the higher multibinding agents, e.g. pentavalent to decavalent compounds.
• the preferred linker length will vary depending upon the distance between adjacent ligand recognition sites , and the geometry, flexibility and composition of the linker.
• the length of the linker will preferably be in the range of about 2-100 Angstroms, more preferably about 2-50 Angstroms, and even more preferably about 5-20 Angstroms.
• Z' and Z " at each separate occurrence are alkylene, cycloalkylene, alkenylene, alkynylene, arylene. heteroarylene, heterocycloalkylene. or a covalent bond; Y' and Y" at each separate occurrence are
• n 0, 1 or 2; and R, R' and R" at each separate occurrence are chosen from hydrogen, alkyl, cycloalkyl. alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl, and heterocyclo.
• linker moiety can be optionally substituted at any atom in the chain by alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, halo, nitro, aryl, heteroaryl, or heterocyclo.
• the simplest (and preferred) construct is a bivalent compound, which can be represented as L-X-L, where L is a ligand that is the same or different at each occurrence, and X is the linker.
• Preferred ligands are triptans. Examples of such compounds are disclosed in the following patents and patent applications, the complete disclosure of which is hereby inco ⁇ orated by reference.
• Preferred ligands may be represented by the following structure:
• R 1 , R 2 , R 6 and R 7 are all hydrogen
• R 3 is hydrogen, heterocyclic, heterocycloalkyl, alkylaminoalkyl. dialkylaminoalkyl, or a linking position;
• R 4 is hydrogen or heterocyclic
• R 5 is hydrogen, fluorine, alkyl, heterocyclic, heteroaryl, heteroarylalkyl, amidoalkyl, alkylaminosulfony lalkyl. dialkylaminosulfonylalkyl, arylsulfonylalkyl, heterocyclosulfonylalkyl, arylcarbonylamino, alkylsulfonamido, or alkylsulfonylalkyl, or a linking position; or R 2 and R 3 when taken together with the carbons to which they are attached represents optionally substituted cycloalkenyl. all of which may be optionally substituted as defined in the Detailed Description ofthe Invention. Particularly preferred are those ligands in which:
• R 3 is dimethylaminoethyl, l-methylpiperidin-4-yl, 1 -methylpyrrolidin-2-yl, methylamino; or a linking position;
• R 5 is methylaminosulfonylmethyl, methylaminosulfonylethyl, l,3-oxazolidin-2-one-3- methyl, 1-triazolemethyl, 4-fluorobenzoylamino, phenylsulfonylethyl, and 1- pyrroiidinsulfonylmethyl;
• R and R when taken together with the carbons to which they are attached represent 4- methylaminocyclohexene.
• sumatriptan is a compound of Formula II in which R , R", R , R and R are all hydrogen, R 3 is (CH 3 ) 2 NCH 2 CH 2 -, and R 5 is CH 3 NHSO 2 CH 2 -, that is:
• naratriptan is a compound of Formula II in which ⁇ S. R- R4 ⁇ R6 an d R7 are all hydrogen, R 3 is N-methylpiperidin-4-yl-, and R 5 is CH 3 NHSO 2 CH 2 -, that is:
• Zolmitriptan is a compound of Formula II in which Rl, R ⁇ , R4, R6 and R? are all hydrogen, R- is (CH ) 2 NCH 2 CH2-, and R ⁇ is oxazolidin-2-one-4-methyl, that is:
• Rizatriptan is a compound of Formula II in which R 1 , R ⁇ , R4, R6 and R 7 are all hydrogen, R 3 is (CH 3 ) NCH 2 CH 2 -. and R 5 is lH-l,2,4-triazole-l -methyl, that is:
• LY-334370 is a compound of Formula II in which R 1 , R 2 , R 4 , R 6 and R 7 are all hydrogen, R J is N-methylpiperidin-4-yl-, and R ⁇ is 4-fluorophenylcarboxamido, that is:
• Eletriptan is a compound of Formula II in which Rl, R2, R4 5 R6 anf j R7 ⁇ g a ⁇ hydrogen, R3 is N-methylpyrrolidine-2-methyl and R ⁇ is 2-(phenylsulfonyl)ethyl, that is:
• Almotriptan is a compound of Formula II in which Rl , R ⁇ , R 4 , R and R 7 are all hydrogen, R- is (CH 3 ) 2 NCH CH2-, and R5 is pyrrolidin-1 -ylsulfonylmethyl, that is:
• VML-251 is a compound of Formula II in which R R4, R6 and R 7 are all hydrogen, R 2 and R 3 when taken together with the carbons to which they are attached represents 4- methylaminocyclohexene, and R ? is carboxamido, that is :
• the triptans may be linked via any position on the ring, including the indole nitrogen.
• Ligands can be linked at a variety of positions using known synthetic methodology.
• An example of positions on a ligand that may be utilized for linking is shown below ( Figure 1), using sumatriptan as an example; the positions available for linking are indicated by arrows.
• the positions indicated by a star are the preferred positions for linking. Additionally, the 3 and 5 positions are preferred for linking in those instances where there is no existing sidechain at the 3 or 5 position.
• ligands it is preferred to link ligands directly, using the functionality already present in the known drug.
• a ring nitrogen e.g., an indole N
• Such intermediates may be commercially available, or are prepared by means well known in the art.
• the intermediates can be linked to each other before the intermediate is modified to the target ligand structure, or linking can occur after such modification.
• the ligand or the ligand intermediate is reacted with a 'core' molecule having two or more functional groups with reactivity that is complementary to that ofthe functional groups on the ligand, thus linking ligands by a linker.
• Selecting different core molecules allows control of linker size, shape, orientation, rigidity, acidity/basicity, hydrophobicity/hydrophilicity, hydrogen bonding characteristics, and number of ligands connected. Examples of "cores" are shown below in Figure 2.
• the solid circle is used in the following figures and reaction schemes to represent any ofthe possible core molecules. That is, the solid circle is equivalent to a linker as defined above after reaction.
• the preferred compounds ofthe invention are bivalent. Accordingly, for the sake of simplicity, the following figures and reaction schemes illustrate the synthesis of bivalent triptans. It should be noted, however, that the same techniques can be used to prepare multibinding agents that are derived from ligands other than triptans, and also generate higher order multibinding compounds, i.e. the compounds ofthe invention where n is 3-10.
• linking at the R 1 position can be accomplished by starting with a monovalent ligand, since the indole N can function as a nucleophile. Using this strategy, the triptan is reacted with a core molecule presenting two or more appropriate electrophilic groups.
• Figure 3 demonstrates this strategy, as applied to sumatriptan, where the electrophilic groups chosen are alkyl bromides.
• the electrophilic groups chosen are alkyl bromides.
• electrophiles equivalent to bromides for example, other halides, mesylates, tosylates, etc.
• Linking at the R " position can also be accomplished by using the monovalent ligand as the starting material, since this position can be functionalized by known techniques. After an electrophilic functional group has been introduced as a sidechain at R 2 , a core molecule presenting two or more appropriate nucleophilic groups, such as amines, is added. A variety of such coupling reactions are well known in the art; examples are shown in Figure 4.
• R 8 is hydrogen , alkyl, alkaryl, alkenyl, alkynyl, cycloalkyl, all of which are optionally substituted as defined in the Detailed Description of the Invention. Linking at the 3-Position
• a preferred method of linking ligands utilizes the sidechains already present on the ligand to form a linker.
• the preferred method for linking these sidechains in many cases requires the preparation of a suitable intermediate.
• linking two triptans through their (CH 3 ) 2 NCH 2 CH 2 - groups (see, for example, sumatriptan) at the R 3 position can be accomplished indirectly through an electrophilic intermediate at the second carbon ofthe sidechain.
• the preparation of such a compound is shown in Figure 5 (the preparation ofthe alcohol intermediate (10) is discussed in more detail in WO 9617842).
• This intermediate is then reacted with a core molecule that has two or more appropriate nucleophilic functional groups.
• a variety of such coupling reactions are well known in the art; Figure 6 illustrates several examples.
• R is hydrogen , alkyl, alkaryl, alkenyl, alkynyl, cycloalkyl, all of which are optionally substituted as defined in the Detailed Description ofthe Invention.
• Linking two ligands through (CH 3 ) 2 NCH 2 CH 2 - groups at the R J position can also be accomplished indirectly by introducing a secondary amine, of the form (CH 3 )HNCH 2 CH2-, into the 3 -position of the triptan as an intermediate.
• a secondary amine of the form (CH 3 )HNCH 2 CH2-
• This intermediate is then reacted with a core molecule having two or more appropriate electrophilic functional groups.
• a variety of such coupling reactions are well known in the art, as shown in Figure 8.
• FIGURE 8 Similar linking reactions may be carried out on ligands having a piperazine substituent at the 3-position, e.g., a ligand derived from naratriptan. for example, having a structure:
• Such a compound is prepared starting from N-(4-fluoro)benzoylation according to a literature procedure (Johnson, Kirk; Phebus, Lee, PCT WO 98/1 1895, 1997).
• ligands exhibit greater variety at the R 3 position, but all can be linked to cores using known chemistry.
• linking two ligands through CH 3 NHSO 2 CH 2 groups can be accomplished by first preparing an electrophilic intermediate of the form (PhO)SO2CH 2 -R.
• the preparation of such an intermediate is shown in Figure 9, and the preparation is described in more detail in DE 3527648 and EP 145459.
• This intermediate is then reacted with a core molecule with two or more appropriate nucleophilic groups, such as amines, as shown in Figure 10.
• FIGURE 10 Where R is hydrogen , alkyl, alkaryl, alkenyl, alkynyl, cycloalkyl, all of which are optionally substituted as defined in the Detailed Description ofthe Invention.
• Linking ligands through an oxazolidinone sidechain can be accomplished directly from the ligand by first protecting the indole nitrogen, as shown in Figure 1 1. After coupling to a core molecule with two or more appropriate electrophilic functional groups, the indole nitrogen is deprotected
• FIGURE 1 A first figure.
• Linking ligands through an oxazolidine sidechain groups can also be accomplished indirectly by first preparing a nucleophilic intermediate ofthe form (NH 2 )(CH 2 OH)CHCH2-R.
• a nucleophilic intermediate ofthe form (NH 2 )(CH 2 OH)CHCH2-R The preparation of such a compound for zolmitriptan is known (ref: Drugs of the Future 1997, 22(3):260-269).
• This intermediate is reacted with a core molecule with two or more appropriate electrophilic functional groups, as shown in Figure 12.
• the oxazolidine ring is formed by known cyclization reactions.
• Another strategy for linking at the 5-position starts from 5-nitroindole, which is commercially available. As shown below in Figure 13. the first step involves substitution at the 3-position by a desired substituent, for example N-methylpiperidine, and then reduction by conventional means to the 5-amino derivative.
• a desired substituent for example N-methylpiperidine
• the compounds of formula (41) and (44), or any similar 5-amino derivative can be converted to a compound of Formula I by reaction of the 5-amino group with a linker via any ofthe routes shown above, e.g., by reacting with .a dicarboxylic acid, a dihalide, a dialdehyde (reductive alkylation), a disulfonyl halide, a diacid halide, a diisocyanate (to give a diureido derivative), and the like.
• the compounds of formula (41) and (44), or any similar 5-amino derivative can be converted to a compound of Formula I that are linked at the 3-position by first reacting the 5-amino group with a desired reagent, for example reaction with 4- fluorobenzoyl chloride to form the desired 5-(4-fluorobenzoylamino) derivative, and then linking the 3-popsition by first removing the Boc protecting group under standard conditions and then reacting the free amino group as described above, e.g., with .a dicarboxylic acid, a dihalide, a dialdehyde (reductive alkylation), a disulfonyl halide, a diacid halide, a diisocyanate (to give a diureido derivative), and the like, to form a bivalent compound.
• a desired reagent for example reaction with 4- fluorobenzoyl chloride to form the desired 5-(4-fluorobenzoylamino) derivative
• linking the 3-popsition by first
• Linking at the 5-positon can also be accomplished by first preparing a 5-carboxy indole derivative, as shown in Figure 15.
• the compounds of formula (45) and (47), or any similar 5-carboxy derivative can be converted to a compound of Formula I by reaction ofthe 5-carboxy group with a linker via any ofthe routes shown above, e.g., by reacting with a diamine. a dihalide, a disulfonyl halide, a diisocyanate (to give a dicarbamate derivative), and the like.
• 5-HT ligands that are not triptans may be prepared and used for linking with triptans.
• a naphthalene derivative can be prepared as shown in Figure 16.
• the compound of formula (48), or any similar hydroxy bearing ligand can be converted to a compound of Formula I in which both ligands are the same by reaction of about 2 molar equivalents of the 5-hydroxy group with about 1 molar equivalent of a linking moiety via any ofthe routes shown above, e.g., by reacting with .a dihalide, a disulfonyl halide, a dicarboxylic acid, and the like.
• a compound of formula (48), or a similar 5- hydroxy derivative is reacted with a large excess of a dihalo compound, for example a dihaloalkane derivative, to give a compound ofthe formula:
• the compound of formula (49) can then be further reacted with a different ligand to provide a compound of Formula I where the ligands are different.
• compounds of Formula I where p is 2 and q is 1 can be prepared in which the 5HT ligands are the same, both being 5HT agonists or both being 5HT antagonists, or are different, in which case both ligands may still both be 5HT agonists or 5HT antagonists, or alternatively a compound of Formula I where one 5HT ligand is a 5HT agonist and the other ligand is a 5HT antagonist may be prepared.
• reaction ofthe compound of formula (49) with a triptan 5HT ligand gives a compound of Formula I with
• reaction of a ligand bearing a 5-amino group with an excess of a dicarboxylic acid, a dihalide, a dialdehyde (reductive alkylation), a disulfonyl halide, a diacid halide, a diisocyanate (to give a diureido derivative), and the like provides a suitably substituted ligand that can be further reacted with a different ligand to provide a compound of Formula I where the ligands are different.
• reaction of a ligand bearing a 5-carboxy derivative with an excess of a diamine, a dihalide, a disulfonyl halide. a diisocyanate provides a suitably substituted ligand that can be further reacted with a different ligand to provide a compound of Formula I where the ligands are different.
• Another strategy, which can be applied to all ligands is to introduce a 'spacer' at any desired position on the molecule before coupling to the central core.
• a spacer can itself be chosen from the possible core compounds, and comprises an electrophile or nucleophile on one end and an electrophile or nucleophile on the other. That is, the ligand is coupled to the spacer, and the product of that reaction is then coupled, after deprotection if necessary, with an appropriate central core. Examples of this type of linking as applied to the 3-position are shown below in Figure 17.
• FIGURE 17 Where R 9 and R 10 are independently chosen from hydrogen, alkyl, alkaryl, alkenyl, alkynyl, cycloalkyl, all of which are optionally substituted as defined in the Detailed Description ofthe Invention.
• All ofthe synthetic strategies described above employ a step in which the ligand, attached to spacers or not, is symmetrically linked to a central core in a single reaction to give a bivalent compound.
• Higher order compounds i.e. multibinding agents in which n is 3-10) can also be synthesized using an asymmetric, linear approach. This strategy is preferred when the target compound is not symmetric.
• Linear synthesis is preferred when linking two or more monomers at different points of connectivity.
• the R 3 of one sumatriptan monomer can be linked to the R 3 or R 1 of another sumatriptan monomer, as shown in Figure 18.
• FIGURE 18 Where R 9 is hydrogen , alkyl, alkaryl, alkenyl. alkynyl, cycloalkyl, all of which are optionally substituted as defined in the Detailed Description of the Invention.
• Type 1 Homodimers Type 3 _ ⁇ ype , Heterodimers
• FIG. 20 A method of preparing multibinding agents where n is 3-10 is illustrated in Figure 20, in which sumatriptan is linked from R to R .
• FIGURE 20 Where R 9 is hydrogen , alkyl, alkaryl, alkenyl, alkynyl, cycloalkyl, all of which are optionally substituted as defined in the Detailed Description of the Invention.
• a linear strategy can also be extended to the synthesis of compounds of Formula I in which L is different. Examples of such reactions are shown in Figure 21, which is a simple extension of the strategies described above.
• FIGURE 21 It should be noted that while all figures are drawn with aliphatic couplings for the sake of simplicity, an alternate linking strategy involves direct substitution on an aryl ring. An example of this is shown in Figure 22.
• Isolation and purification ofthe compounds and intermediates described herein can be effected, if desired, by any suitable separation or purification procedure such as, for example, filtration, extraction, crystallization, column chromatography, thin-layer chromatography, thick-layer chromatography, preparative low or high-pressure liquid chromatography or a combination of these procedures.
• suitable separation and isolation procedures can be had by reference to the Examples hereinbelow. However, other equivalent separation or isolation procedures could, of course, also be used.
• DIPEA diisopropylethylamine
• Hunig's base DMA N,N-dimethylacetamide
• DPPA diphenylphosphoryl azide HATU 6-(7-azabenzotriazol- 1 -y -N.N.N" ⁇ tetramethyluronium hexafluorophosphate
• reaction mixtures were worked up as described specifically in each reaction; normally it was purified by flash column chromatography with silica gel.
• reaction mixtures were routinely purified by preparative HPLC: a general protocol is described below. Characterization of reaction products was routinely performed by IH-NMR spectrometry; samples were dissolved in deuterated solvent (CD 3 OD, CDC1 3 . or DMSO-d 6 ), and followed by acquisition of its IH-NMR spectra with a Varian Gemini 2000 instrument (300 MHz) under standard observe parameters. Mass spectrometric identification of compounds was performed by an electrospray ionization method (ESMS) with a Perkin Elmer instrument (PE SCIEX API 150 EX).
• ESMS electrospray ionization method
• PE SCIEX API 150 EX Perkin Elmer instrument
• reaction of 1 molar equivalent each of two dissimilar ligands bearing an amino group with 1 molar equivalent of a linker having two chemically complementary groups forms a compound of Formula I having two different ligands (a heterodimer).
• reaction of 1 molar equivalent of a ligand bearing an amino group with 1 molar equivalent of a linker having two chemically complementary groups followed by reaction ofthe intermediate thus formed with a second (and diferent) ligand provides a compound of Formula I that is a heterodimer.
• reaction mixture was cooled to 0°C, followed by dropwise addition of 2M MeNH 2 in THF (400 mL). After addition, the mixture was warmed gradually to room temperature, and stirred for 2 h. The reaction mixture was concentrated in vacuo, yielding a pale yellow solid residue. It was partitioned between water (300 mL) and CH2CI 2 (500 mL). and the organic phase was removed. Solid mass in the aqueous phase was collected on Buchner funnel, and washed with water (300 mL), 1.0 M NaOH (200 mL). water (300 mL), and MeOH (500 mL).
• reaction of 1 molar equivalent of a ligand bearing an amino group with 1 molar equivalent of a linker having two chemically complementary groups followed by reaction ofthe intermediate thus formed with a second (and diferent) ligand provides a compound of Formula I that is a heterodimer.
• reaction of 1 molar equivalent of a ligand bearing an amino group with 1 molar equivalent of a linker having two chemically complementary groups followed by reaction ofthe intermediate thus formed with a second (and diferent) ligand provides a compound of Formula I that is a heterodimer.
• reaction of 1 molar equivalent of a ligand bearing an amino group with 1 molar equivalent of a linker having two chemically complementary groups followed by reaction ofthe intermediate thus formed with a second (and diferent) ligand provides a compound of Formula I that is a heterodimer.
• the reaction is allowed to cool, and is then quenched with aqueous NH 4 C1 solution until the pH ofthe solution is adjusted to pH 7.0 using either 1 M HCl or 1 M NaOH.
• the product is extracted from this aqueous phase with EtOAc.
• the organic layer is dried using Na SO 4 , the drying agent is then filtered off and the solvent removed in vacuo to provide the crude product.
• the desired material is .purified from this mixture using reverse phase HPLC.
• reaction is quenched with saturated aqueous NH 4 C1 solution and extracted with EtOAc.
• organic layer is dried over Na 2 SO 4 , the drying agent is then filtered off, and the solvent removed in vacuo to provide the crude product.
• the desired material is then purified from this mixture using reverse phase HPLC.
• This product (protected zolmitriptan) (387 mg, 1 mmol), is then dissolved in DMF (10 ml) and cooled to 0°C. NaH (27 mg. 1.1 mmol) is then added to the cold solution, then the reaction vessel is allowed to warm to room temperature and stirred for 30 minutes.
• a solution of 1 ,4-bis(bromomethyl)benzene, a compound of formula (2) (132 mg, 0.5 mmols) in DMF (10 ml) is added dropwise to the reaction vessel. After 2 hours, the reaction is quenched with saturated aqueous NaHCO 3 , and extracted into EtOAc. The organic layer is then dried using Na 2 SO 4 , the drying agent is then filtered off, and the solvent removed in vacuo to provide the crude product. The desired material is then purified from this mixture using reverse phase HPLC.
• This product (protected zolmitriptan dimer) 19 (677 mg, 1 mmol), is dissolved in CH 2 C1 2 (4 ml). A solution of 10% trifluoroacetic acid in CH 2 C1 2 (10 ml) is added, and the reaction is stirred for 1 hour at room temperature. The solvent is then remove under vacuum to provide the desired material as the TFA salt. The desired material is then purified from this mixture using reverse phase HPLC.
• the selective 5-HT I F receptor agonist can be used as a standard in these assays as it displaces [ 3 H] sumatriptan binding to 5- HT I F receptors with nanomolar affinity.
• a similar assay using a radiolabelled version of a 5HT I selective ligand such as LY 334370 can also be used.
• 5-HTI B Receptor Agonists In vitro saphenous venoconstrictor assay The ability of compounds to contract the isolated saphenous vein of the rabbit or dog is used to determine the potency and efficacy of novel 5-HT ] B receptor agonists (Connor et al., 1997; Martin, 1997). As both the saphenous vein and cranial vasculature contain contractile 5-HT ⁇ B receptors, the former tissue is used as a marker for cranial vasoconstrictor activity of 5-HT I B receptor agonists.
• Rings of the rabbit saphenous vein are mounted in organ baths containing a physiological Krebs solution and drug-induced changes in tension are recorded. The potency and efficacy of compounds is measured.
• Naratriptan biological profile in animal models relevant to migraine. Cephalalgia, 17, 145-152.
• the selective 5-HT ⁇ receptor agonists that are clinically effective anti-migraine agents have little affinity for 5-HT A receptors (e.g. sumatriptan; Humphrey et al., 1988).
• the rabbit isolated thoracic aorta is used to determine the 5-HT 2A activity of novel compounds.
• Thoracic aorta rings are mounted in organ baths containing a physiological Krebs solution, and drug-induced changes in tension are recorded. The potency and efficacy of compounds is measured
• Butina D. (1988). GR43175, a selective agonist for the 5-HT ⁇ -like receptor in dog isolated saphenous vein. Br. J. Pharmacol, 94, 1123-1 132. Krisch, I., Budihna, M.V. and Rucman, R. (1992). Structure-activity study of some newly synthesized ergoline derivatives on 5-HT2 receptors and alpha-adrenoceptors in rabbit isolated aorta. Pharmacol., 45, 195-208
• Trigeminal nerve stimulation produces inflammation (consisting of plasma protein extravasation) in cranial tissues, an event implicated in migraine pathogenesis.
• 5-HT I B/I D/I F receptors inhibits plasma protein extravasation in the dura of anesthetized rats and guinea-pigs following either electrical or chemical trigeminal nerve stimulation (Buzzi and Moskowitz, 1992; Phebus et al., 1997; Shepheard et al., 1997). It has been suggested that the inhibitory activity of 5-HT ⁇ receptor agonists (e.g. sumatriptan) on neurogenic inflammation contributes to their anti-migraine efficacy.
• 5-HT ⁇ receptor agonists e.g. sumatriptan
• guinea-pigs or rats are anesthetized and placed in a stereotaxic frame. The skull is exposed by a midline incision and a hole is drilled on either side for electrode placement into each trigeminal ganglion. Novel compounds are administered, followed by 125 I-bovine serum albumin. One trigeminal ganglion is electrically stimulated, and animals are then perfused with saline via the left cardiac ventricle. The right atrium is incised to allow outflow of perfusate. The eyelid, eyeball and dura are dissected out. Tissues from the stimulated and unstimulated sides are weighed and counted for radioactivity in a gamma counter.
• the potency of compounds in inhibiting neurogenic cranial inflammation is determined.
• I-bovine serum albumin As an alternative to I-bovine serum albumin, Evan's Blue (50mg/kg i.v.), detected by fluorescence microscopy or HPLC, can be used to measure the plasma protein extravasation evoked by trigeminal ganglion stimulation.
• EXAMPLE 21 In vitro 5-HT I B /I D receptor binding assay The 5-HT I B/I D binding affinity and selectivity of compounds is measured in radioligand displacement assays using commercially available membrane preparations (Euroscreen s.a., Brussels/Belgium) from a CHO-K1 cell line expressing the cloned human 5-HT I D and 5-HT I B receptors.
• Radioligand displacement assays are carried out at a constant concentration ofthe radioligand [ ⁇ ]5-hydroxytryptamine with increasing concentrations (10 " nding reactions are incubated for 1 hour at room temperature in a buffer of 50mM Tris-HCl pH 7.4, ImM EDTA, 12.5mM MgCl 2 , 0.1%) ascorbic acid and are stopped by rapid filtration over GF/B glassfiber filters using a cell harvester. Filter-bound radioactivity is counted in a liquid scintillation counter.
• the selective 5-HT I D agonists sumatriptan and rizatriptan may be used as standards in these assays.
• EXAMPLE 22 This example illustrates the preparation of a representative pharmaceutical formulation for oral administration containing a multibinding compound of the invention. Ingredients Quantity per tablet, mgs.
• the above ingredients are mixed and introduced into a hard-shell gelatin capsule.
• multibinding compounds ofthe invention can be used as the active compound in the preparation ofthe orally administrable formulations of this example.
• This example illustrates the preparation of another representative pharmaceutical formulation for oral administration containing a multibinding compound of the invention.
• EXAMPLE 24 This example illustrates the preparation of a representative pharmaceutical formulation containing a multibinding compound ofthe invention
• An oral suspension is prepared having the following composition. Ingredients
• Veegum K (Vanderbilt Co.) 1.0 g
• multibinding compounds ofthe invention can be used as the active compound in the preparation of the orally administrable formulations of this example.
• This example illustrates the preparation of a representative pharmaceutical formulation containing a multibinding compound ofthe invention.
• An injectable preparation buffered to a pH of 4 is prepared having the following composition: Ingredients
• EXAMPLE 26 This example illustrates the preparation of a representative pharmaceutical formulation for injection containing a multibinding compound ofthe invention.
• a reconstituted solution is prepared by adding 20 ml of sterile water to lg ofthe compound of Formula I. Before use, the solution is then diluted with 200 ml of an intravenous fluid that is compatible with the compound of Formula I.
• Such fluids are chosen from 5% dextrose solution, 0.9% sodium chloride, or a mixture of 5% dextrose and 0.9% sodium chloride.
• Other examples are lactated Ringer's injection, lactated Ringer's plus 5% dextrose injection. Normosol-M and 5% dextrose, Isolyte E. and acylated Ringer's injection
• EXAMPLE 27 This example illustrates the preparation of a representative pharmaceutical formulation for topical application containing a multibinding compound ofthe invention. Ingredients grams
• multibinding compounds of the invention can be used as the active compound in the preparation of topical formulations of this example.
• This example illustrates the preparation of a representative pharmaceutical formulation containing a multibinding compound ofthe invention.
• a suppository totalling 2.5 grams is prepared having the following composition: Ingredients
• Active Compound 500 mg Witepsol H-15 balance * triglycerides of saturated vegetable fatty acid; a product of Riches-Nelson, Inc., New York, N.Y.

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