Therapeutic compounds for pain and their synthesis

专利摘要:

公开号:ES2809703T9
申请号:ES16842971T
申请日:2016-09-01
公开日:2022-01-24
发明作者:Bart Lieven Daniel Decorte；Jacob Cornelis Russcher；Menno Cornelis Franciscus Monnee
申请人:Janssen Pharmaceutica NV；
IPC主号:

专利说明:

[0002] Therapeutic compounds for pain and their synthesis
[0004] field of invention
[0006] The invention provides new pharmaceutically active chemical compounds, which can be used to treat conditions and disorders in animals, mammals and humans.
[0008] Background
[0010] New chemical compounds that have pharmaceutical activity may be indicated for the treatment of previously untreatable conditions, better treatment of conditions than can be achieved with conventional pharmaceuticals, and treatment of conditions that were previously treatable with conventional pharmaceuticals, but now it can no longer be treated effectively. For example, such compounds may be useful in the case of bacterial or viral infectious agents that have evolved drug resistance.
[0012] WO 92/06188 relates to quinuclidine derivatives and pharmaceutically acceptable salts thereof. The compounds are said to be antagonists of substance P and are therefore said to be useful in the treatment of gastrointestinal disorders, central nervous system disorders, inflammatory disorders, pain and migraine.
[0014] WO2007/024814 relates to amide and urea derivatives of heteroaryl-substituted diazatricycloalkanes, pharmaceutical compositions including the compounds, methods of preparing the compounds, and methods of treatment using the compounds. More specifically, methods of treatment are said to involve modulating the activity of the a7 nAChR subtype by administering one or more of the compounds to treat or prevent disorders mediated by the a7 nAChR subtype. Diazatricycloalkanes typically consist of a 1-azabicyclooctane fused to the pyrrolidine ring.
[0016] WO2008/112734 relates to heterocyclo-carbonyl-diazabicycloalkanes as putative modulators of the neural nicotinic acetylcholine alpha-4 beta subtype receptor for the treatment of CNS-related disorders.
[0018] SUMMARY OF THE INVENTION
[0020] The invention provides a compound of Formula 1:
[0025] In certain embodiments, Formula 1 is
[0028] ((2S*,3S*,3aS*,6R*,7aR*)-1-(pyridin-2-ylmethyl)-4-(3,3,3-trifluoropropanoyl)octahydro-1H-2,6-methanepyrrolo[3 ,2-b]pyridin-3-yl pivalate).
[0030] In certain embodiments, the invention includes a pharmaceutical composition containing a compound of Formula 1 and/or a derivative thereof. In one embodiment, the invention includes a pharmaceutical composition comprising a compound of Formula 1 and/or a derivative thereof and a pharmaceutically acceptable carrier or diluent. In another embodiment, the invention provides a method of treating a subject (a human or an animal) suffering from a condition, disease or disorder, comprising administering to the subject an effective amount of a compound of Formula 1 and/or derivative thereof . In one embodiment, the compound is administered to effect localized administration to the subject. In another embodiment, the compound is administered to effect systemic administration to the subject. In a further embodiment, a compound of Formula 1, and/or a derivative thereof is used as a medicament, or is used in the manufacture of a medicament. In some embodiments, the condition or disorder is neuropathic pain or chronic pain.
[0032] In other embodiments, the method includes making the compound of Formula 1. In one such embodiment, the method of making the compound of Formula 1 includes reacting a compound of Formula 2:
[0037] (2S*,3S*,3aS*,6R*,7aR*)-4-(3,3,3-trifluoropropanoyl)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridin-3-yl pivalate with 2-pyridinecarboxaldehyde in the presence of a reducing agent. In some embodiments, the 2-pyridinecarboxaldehyde was added before the reducing agent. In certain embodiments, the reducing agent is sodium triacetoxyborohydride. In some embodiments, the compound of Formula 1 is chirally separated.
[0039] In some embodiments, the method may also include making the compound of Formula 2. In one embodiment, the method of making the compound of Formula 2 includes reacting a compound of Formula 3:
[0044] (2S*,3S*,3aS*,6R*,7aR*)-tert-butyl 3-(pivaloyloxy)-4-(3,3,3-trifluoropropanoyl)octahydro-1H-2,6-methanepyrrolo[3,2 -b]pyridine-1-carboxylate with an acid. In certain embodiments, the acid is trifluoroacetic acid.
[0046] In some embodiments, the method may also include making the compound of Formula 3. In one embodiment, the method of making the compound of Formula 3 includes reacting a compound of Formula 4:
[0049] (2S*,3S*,3aS*,6R*,7aR*)-tert-butyl 3-hydroxy-4-(3,3,3-trifluoropropanoyl)octahydro-1H-2,6-methanepyrrolo[3,2-b ]pyridine-1-carboxylate with dimethylaminopyridine (DMAP).
[0050] In some embodiments, the method may also include making the compound of Formula 4. In one embodiment, the method of making the compound of Formula 4 includes reacting a compound of Formula 5:
[0055] (2S*,3S*,3aS*,6R*,7aR*)-tert-butyl 3-((tert-butyldiphenylsilyl)oxy)-4-(3,3,3trifluoropropanoyl)octahydro-1H-2,6-methanepyrrolo[3, 2-b]pyridine-1-carboxylate with tert-butyldiphenylchlorosilane. In some embodiments, the reaction further comprises pyridine.
[0056] In some embodiments, the method may also include making the compound of Formula 5. In one embodiment, the method of making the compound of Formula 5 includes reacting a compound of Formula 6.b:
[0060] (2S*,3R*,3aS*,6R*,7aR*)-tert-butyl 3-((tert-butyldiphenylsilyl)oxy)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1-carboxylate with 3,3,3-trifluoropropanoic acid. In some embodiments, the reaction further comprises NN-diisopropylethylamine. In certain embodiments, the reaction further comprises (1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate)sodium triacetoxyborohydride. In some embodiments, the method includes chirally separating a compound of Formula 7:
[0063] rac-(2S*,3R*,3aS*,6R*,7aR*)-tert-butyl 3-((tert-butyldiphenylsilyl)oxy)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1- carboxylate.
[0065] In other embodiments, the method includes making the compound of Formula 7. In one such embodiment, the method of making the compound of Formula 7 includes reacting a compound of Formula 8:
[0070] rac-(2R,3R,6S,7aS)-tert-butyl 4-benzyl-3-((tert-butyldiphenylsilyl)oxy)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1-carboxylate with hydrogen. The reaction can be carried out in the presence of a catalyst. In a preferred embodiment, the catalyst includes palladium. For example, the catalyst can be palladium on carbon.
[0072] In other embodiments, the method includes making the compound of Formula 8. In one such embodiment, the method of making the compound of Formula 8 includes reacting a compound of Formula 9:
[0077] rac-(2R,3R,6S,7aS)-4-benzyl-3-((tert-butyldiphenylsilyl)oxy)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine with di-tert-dicarbonate butyl (Boc ² O) to add a tert-butyloxycarbonyl (Boc) protecting group. In a preferred embodiment, the reaction further comprises triethylamine (Et ³ N).
[0079] In other embodiments, the method also includes making the compound of Formula 9. In one such embodiment, the method of making the compound of Formula 9 includes reacting a compound of Formula 10:
[0081] (2R,3R,6S,7aS)-ethyl 4-benzyl-3-((tert-butyldiphenylsilyl)oxy)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1-carboxylate with iodotrimethylsilane.
[0082] In other embodiments, the method also includes making the compound of Formula 10. In one such embodiment, the method of making the compound of Formula 10 includes reacting a compound of Formula 11:
[0086] (2R,3S,6S,7aS)-ethyl-4-benzyl-3-hydroxyoctahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1-carboxylate with TBDPS. In a preferred embodiment, the reaction further comprises imidazole.
[0087] In other embodiments, the method also includes making the compound of Formula 11. In one such embodiment, the method of making the compound of Formula 11 includes reacting a compound of Formula 12:
[0091] (2R,3S,6S,7aS)-ethyl 3-hydroxyoctahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1-carboxylate with benzaldehyde. In a preferred embodiment, the reaction further comprises sodium triacetoxyborohydride (STAB).
[0092] In other embodiments, the method also includes making the compound of Formula 12. In one such embodiment, the method of making the compound of Formula 12 includes cyclizing a compound of Formula 12.a:
[0095] (1R,2R,4S,5S,7s)-ethyl 7-(aminomethyl)-3-oxa-9-azatricyclo[3.3.1.02.4]nonane-9-carboxylate in a solvent. The solvent can be ethanol (EtOH).
[0096] In other embodiments, the method also includes making the compound of Formula 12.a. In one such embodiment, the method of making the compound of Formula 12.a includes reacting a compound of Formula 13:
[0101] (1R,2R,4S,5S,7s)-ethyl 7-cyano-3-oxa-9-azatricyclo[3.3.1.02.4]nonane-9-carboxylate with hydrogen. The reaction can be carried out in the presence of a catalyst. In one embodiment, the catalyst includes nickel. For example, the catalyst can be Raney nickel.
[0102] In other embodiments, the method also includes making the compound of Formula 13. In one such embodiment, the method of making the compound of Formula 13 includes reacting a compound of Formula 14:
[0107] (1R,2R,4S,5S,7r)-ethyl 7-((methylsulfonyl)oxy)-3-oxa-9-azatricyclo[3.3.1.02.4]nonane-9-carboxylate with potassium cyanide. In other embodiments, the reaction further comprises 18-crown-6 (1,4,7,10,13,16-hexaoxacyclooctadecane).
[0108] In other embodiments, the method also includes making the compound of Formula 14. In one such embodiment, the method of making the compound of Formula 14 includes reacting a compound of Formula 15:
[0113] (1R,2R,4S,5S,7r)-ethyl 7-hydroxy-3-oxa-9-azatricyclo[3.3.1.02.4]nonane-9-carboxylate with mesyl chloride. In a preferred embodiment, the reaction further comprises triethylamine (ET3N).
[0114] In other embodiments, the method also includes making the compound of Formula 15. In one such embodiment, the method of making the compound of Formula 15 includes reacting a compound of Formula 16:
[0117] (1R,2R,4S,5S,7r)-ethyl 7-(benzoyloxy)-3-oxa-9-azatricyclo[3.3.1.02.4]nonane-9-carboxylate with a reducing agent. The reducing agent may be sodium borohydride.
[0119] In other embodiments, the method also includes making the compound of Formula 16. In one such embodiment, the method of making the compound of Formula 16 includes reacting a compound of Formula 17:
[0124] (1R,2R,4S,5S,7r)-9-methyl-3-oxa-9-azatricyclo[3.3.1.02.4]nonan-7-yl benzoate with ethyl chloroformate. In a preferred embodiment, the reaction further comprises a base. The base can be potassium carbonate.
[0126] In other embodiments, the method also includes making the compound of Formula 17. In one such embodiment, the method of making the compound of Formula 17 includes reacting a compound of Formula 18:
[0131] (1R,2R,4S,5S)-9-methyl-3-oxa-9-azatricyclo[3.3.1.02.4]nonan-7-ol) with benzoic acid in the presence of an activating agent. The activating agent can be diethylazodicarboxylate (DEAD) with triphenylphosphine (PPh3) or diisopropyl azodicarboxylate (DIAD) with PPh3.
[0133] In other embodiments, the method also includes making the compound of Formula 18. In one such embodiment, the method of making the compound of Formula 18 includes reacting a compound of Formula 19:
[0138] (2S)-(1 R,2R,4S,5S)-9-methyl-3-oxa-9-azatricyclo[3.3.1.02.4]nonan-7-yl-3-hydroxy-2-phenylpropanoate hydrobromide trihydrate (scopolamine ) with a reducing agent The reducing agent may be sodium borohydride. In a preferred embodiment, the reaction further comprises HCl in isopropyl alcohol.
[0140] In some embodiments, the compounds described herein are used in the treatment or prevention of neuropathic pain in a subject in need thereof. In other embodiments, the compounds described herein are useful in the treatment or prevention of chronic pain in a subject in need thereof.
[0142] BRIEF DESCRIPTION OF THE DRAWINGS
[0144] The foregoing summary, as well as the following detailed description of the invention, can be better understood when read in conjunction with the accompanying Figures. For the purpose of illustrating the invention, the Figures show embodiments of the present invention. However, it is to be understood that the invention is not limited to the precise arrangements, examples and instruments shown.
[0146] Figure 1 shows the results of a 1HNMR (CDCb) analysis of the compound of Formula 18, according to one embodiment of the invention.
[0147] Figure 2 shows the results of an MS analysis of the compound of Formula 17, according to one embodiment of the invention.
[0148] Figures 3A and 3B show the results of a structural analysis of the compound of Formula 16. Figure 3A shows the results of a 1HNMR analysis of the compound of Formula 16. Figure 3B shows the results of an MS analysis of the compound of Formula 16. .
[0149] Figure 4 shows the results of a 1HNMR analysis of the compound of Formula 15.
[0150] Figure 5 shows the results of a 1HNMR analysis of the compound of Formula 14.
[0151] Figure 6 shows the results of a 1HNMR analysis of the compound of formula 13.
[0152] Figure 7 shows the results of a 1HNMR analysis of the compound of formula 12.
[0153] Figures 8A and 8B show the results of a structural analysis of the compound of Formula 11. Figure 8A shows the results of an MS analysis of the compound of Formula 11. Figure 8B shows the results of a 1HNMR analysis of the compound of Formula eleven.
[0154] Figures 9A and 9B show the results of a structural analysis of the compound of Formula 10. Figure 9A shows the results of an LCMS analysis of the compound of Formula 10. Figure 9B shows the results of a 1HNMR analysis of the compound of Formula 10.
[0155] Figure 10 shows the results of an LCMS analysis of the compound of Formula 9.
[0156] Figures 11A and 11B show the results of a structural analysis of the compound of Formula 8. Figure 11A shows the results of a 1HNMR analysis of the compound of Formula 8. Figure 11B shows the results of an LCMS analysis of the compound of Formula 8.
[0157] Figures 12A and 12B show the results of a structural analysis of the compound of Formula 7. Figure 12A shows the results of an LCMS analysis of the compound of Formula 7. Figure 12B shows the results of a 1HNMR analysis of the compound of Formula 7.
[0158] Figure 13 shows the results of a 1HNMR analysis of the compound of Formula 2.
[0159] Figure 14 shows the results of a 1HNMR analysis of the compound of Formula 1.
[0160] Figure 15 shows the effect of daily administration of Formula 1 from day 8 to day 14 on the expression of 14-3-3a in the blood of mice dosed with Taxol, according to one embodiment of the invention.
[0161] Figure 16 demonstrates changes in 14-3-3a in the posterior plantar skin of mice administered Taxol and treated with Formula 1. As illustrated, 14-3-3a expression was significantly reduced after repeated dosing with Formula 1 vs. vehicle (p < 0.01), according to one embodiment of the invention.
[0162] Figure 17 demonstrates changes in Formula 1 following the IV route of administration at a dose of 5 mg/kg or the PO route of administration at a dose of 50 mg/kg. Plasma collection was performed over 8 hours, according to one embodiment of the invention.
[0163] Figure 18 shows the level of Formula 1 in the feces of mice after IV (5 mg/kg) and PO (50 mg/kg) dosing, according to one embodiment of the invention.
[0164] Figure 19 shows the activity of Formula 1 in Taxol-induced neuropathic pain, according to one embodiment of the invention.
[0165] Figure 20 illustrates the activity of Formula 1 in CCI-induced neuropathic pain, according to one embodiment of the invention.
[0166] Figure 21 illustrates the activity of Formula 1 on cold allodynia after CCI-induced neuropathic pain, according to one embodiment of the invention.
[0167] Figure 22 illustrates the activity of Formula 1, dosed orally, on Taxol-induced neuropathic pain in mice, according to one embodiment of the invention.
[0168] Figure 23 illustrates the activity of Formula 1, using the oral route of administration, on Taxol-induced neuropathic pain in rats, according to one embodiment of the invention.
[0169] Figure 24 illustrates the effect of Formula 1 treatment on body weight loss after Taxol dosing.
[0170] Figure 25 illustrates the effect of Formula 1 in a rat model of CCI, according to one embodiment of the invention. Continuous dosing of formula 1 resulted in an increase in the activity of the compound.
[0171] Detailed description
[0172] Embodiments of the invention are discussed in detail below. In describing these embodiments, specific terminology is used for the sake of clarity. However, the invention is not intended to be limited to the specific terminology selected.
[0173] certain definitions
[0174] The term "alkyl" refers to branched or unbranched hydrocarbon chains, for example, hydrocarbon chains having from 1 to 12 carbon atoms in the chain. In some embodiments, an alkyl group is a C ¹ -C ⁶ alkyl group. In some embodiments, an alkyl group is a C ¹ -C ⁴ alkyl group. Examples of alkyl groups include methyl (Me), ethyl (Et), n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl (tBu), pentyl, isopentyl, tert-pentyl, hexyl, isohexyl, and which in light of ordinary skill in the art and the teachings provided herein would be considered equivalent to any of the foregoing examples.
[0175] The term "haloalkyl" refers to a straight or branched chain alkyl group having from 1 to 12 carbon atoms in the chain and having at least one of the hydrogens replaced by a halogen. In some embodiments, a haloalkyl group is a C ¹ -C ⁶ haloalkyl group. In some embodiments, a haloalkyl group is a C ¹ -C ⁴ haloalkyl group. An exemplary substituent is fluorine. Preferred substituted alkyl groups of the invention include trihalogenated alkyl groups such as trifluoromethyl groups. Haloalkyl includes and is not limited to CF ³ , CH ² F, -CHF ² , -CH ² Cl, -CH ² -CF ³ and the like.
[0176] "Cycloalkyl" refers to monocyclic nonaromatic hydrocarbon groups having from 3 to 7 carbon atoms. Examples of cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
[0177] The term "alkoxy" includes a straight or branched chain alkyl group with a terminal oxygen linking the alkyl group to the rest of the molecule. In some embodiments, an alkoxy group is a C ¹ -C ⁶ alkoxy group. In some embodiments, an alkoxy group is a C ¹ -C ⁴ alkoxy group. Alkoxy includes methoxy, ethoxy, propoxy, isopropoxy, butoxy, t-butoxy, pentoxy, and so on.
[0178] The term "heterocycle" represents a mono- or bicyclic hydrocarbon ring structure optionally containing heteroatoms selected from O, S and N. Heterocyclyl rings may have from 2 to 10 carbon ring atoms.
[0179] The term "halogen" represents chlorine, fluorine, bromine or iodine. The term "halo" represents chlorine, fluorine, bromine or iodine.
[0180] A wavy line ' ' indicates the point of attachment to the rest of the molecule.
[0181] "Benzyl" and -CH ² -phenyl are used interchangeably.
[0182] "Pharmaceutically acceptable" means approved or may be approved by a federal or state government regulatory agency or the appropriate agency in countries other than the United States, or that is listed in the United States Pharmacopeia or other generally recognized pharmacopeia for its use in animals, and more particularly, in humans.
[0183] "Pharmaceutically acceptable salt" refers to a salt of a compound of the invention that is pharmaceutically acceptable and that possesses the desired pharmacological activity of the parent compound. In particular, such non-toxic salts may be inorganic or organic acid addition salts and base addition salts. Specifically, such salts include: (1) acid addition salts, formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or formed with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid , 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-Toluenesulfonic Acid, Camphorsulfonic Acid, 4-Methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic Acid, Glucoheptonic Acid, 3-Phenylpropionic Acid, Trimethylacetic Acid, tertiary-Butylacetic Acid, Laurylsulfuric Acid, Gluconic Acid, Acid glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid and the like; or (2) salts formed when an acidic proton present in the parent compound is replaced by a metal ion, eg, an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine, and the like. Salts further include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and when the compound contains a basic functionality, salts of non-toxic organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate and the like.
[0184] "Pharmaceutically acceptable carrier" refers to a diluent, adjuvant, excipient, or carrier with which a compound of the invention is administered. A "pharmaceutically acceptable carrier" refers to a substance that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to a subject, as an inert substance, added to a pharmacological composition, or otherwise used as a carrier. , carrier or diluent to facilitate the administration of an agent and that is compatible therewith. Examples of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.
[0185] "Subject" includes humans. The terms "human", "patient" and "subject" are used interchangeably herein.
[0186] "Treat" or "treatment" of any disease or disorder refers, in one embodiment, to ameliorate the disease or disorder (i.e., stop or reduce the development of the disease or at least one of the clinical symptoms thereof) . In another embodiment, "treating" or "treatment" refers to improving at least one physical parameter, which may not be discernible by the subject. In yet another embodiment, "treating" or "treatment" refers to modulating the disease or disorder, either physically (eg, stabilization of a discernible symptom), physiologically (eg, stabilization of a physical parameter), or both. In yet another embodiment, "treating" or "treatment" refers to delaying the onset of the disease or disorder.
[0187] In methods of treatment according to the invention, a therapeutically effective amount of a pharmaceutical agent according to the invention is administered to a subject suffering from or diagnosed with such a disease, disorder or condition. A "therapeutically effective amount" means an amount or dose sufficient to generally achieve the desired therapeutic or prophylactic benefit in patients in need of such treatment for the designated disease, disorder, or condition.
[0188] Effective amounts or doses of the compounds of the present invention can be determined by routine methods such as modeling, dose escalation studies, or clinical trials, and by taking into account routine factors, for example, the mode or route of drug administration or administration, the pharmacokinetics of the compound, the severity and course of the disease, disorder or condition, the subject's prior or ongoing therapy, the subject's health status and response to drugs, and the judgment of the treating physician. An example of a dose is in the range of from about 0.001 to about 200 mg of compound per kg of body weight of the subject per day, preferably from about 0.05 to 100 mg/kg/day, or from about 1 to 35 mg/kg. kg/day, in single or divided dosage units (eg BID, TID, QID). For a 70 kg human, an illustrative range for a suitable dosage amount is from about 0.05 to about 7 g/day, or from about 0.2 to about 2.5 g/day.
[0189] "Compounds of the present invention", and equivalent expressions, are intended to encompass compounds of the Formula as described herein, which expression includes pharmaceutically acceptable salts and solvates, eg, hydrates, where the context permits. Similarly, reference to intermediates, whether or not they themselves are claimed, is intended to encompass their salts and solvates, where the context permits.
[0190] As used herein, the term "isotopic variant" refers to a compound that contains unnatural ratios of isotopes at one or more of the atoms that constitute said compound. For example, an "isotopic variant" of a compound may be radiolabeled, that is, contain one or more nonradioactive or radioactive isotopes, such as deuterium (2H or D), carbon-13 (13C), nitrogen-15 ( 15N), or the like. It will be understood that, in a compound where such isotopic substitution is made, the following atoms, when present, may vary so that, for example, any hydrogen may be 2H/D, any carbon may be 13C, or any nitrogen may be 15N, and that the presence and placement of such atoms can be determined within the skill of the art. Similarly, the invention may include the preparation of isotopic variants with radioisotopes, in the case, for example, where the resulting compounds may be used for drug and/or substrate tissue distribution studies. The radiolabeled compounds of the invention can be used in diagnostic methods such as single photon emission computed tomography (SPECT). The radioactive isotopes tritium, ie, 3H, and carbon-14, ie, 14C, are particularly useful for their ease of incorporation and ready means of detection. In addition, compounds that are substituted with positron emitting isotopes such as 11C, 18F, 15O, and 13N can be prepared, and will be useful in Positron Emission Tomography (PET) studies to examine substrate receptor occupancy.
[0191] All isotopic variants of the compounds of the invention, radioactive or not, are intended to be embraced within the scope of the invention. In one aspect, deuterated or tritiated analogs of the disclosed compounds are provided herein.
[0192] It should also be understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are called "isomers". Isomers that differ in the arrangement of their atoms in space are called "stereoisomers".
[0193] Stereoisomers that are not mirror images of each other are called "diastereomers" and those that are non-superimposable mirror images of each other are called "enantiomers". When a compound has an asymmetric center, for example, it is attached to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the way the molecule rotates the plane of polarized light and is designated dextrorotatory or levorotatory (ie as (+) or (-) isomers respectively). A chiral compound can exist as a single enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a "racemic mixture".
[0194] "Tautomers" refer to compounds that are interchangeable forms of a particular compound structure, and that vary in the displacement of hydrogen atoms and electrons. Thus, two structures can be in equilibrium through the movement of n electrons and one atom (usually H). For example, enols and ketones are tautomers because they rapidly interconvert upon treatment with acid or base. Another example of tautomerism is the aci- and nitro- forms of phenyl nitromethane, which are formed in the same way by treatment with acid or base.
[0195] Tautomeric forms may be relevant to achieving optimal chemical reactivity and biological activity of a compound of interest.
[0196] Compounds of the invention may also exist as "rotamers", i.e., conformational isomers that occur when rotation leading to different conformations is hindered, resulting in a rotational energy barrier that must be overcome to convert from one conformational isomer to another. other.
[0197] The compounds of this invention may possess one or more asymmetric centers; such compounds may therefore occur as individual (R)- or (S)-stereoisomers or as mixtures thereof.
[0198] Unless otherwise indicated, the description or designation of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic or otherwise, thereof. Methods for stereochemistry determination and separation of stereoisomers are well known in the art.
[0199] As used herein, the term "localized administration" denotes the administration of a pharmaceutical or therapeutic agent to a specific, limited region of the body.
[0200] As used herein, the term "systemic administration" denotes administration of a pharmaceutical or therapeutic agent throughout the body, for example, via administration to the circulatory system.
[0201] As used herein, the term "mass spectrometry (MS)" denotes an analytical technique that ionizes a chemical compound to generate charged molecules or molecule fragments and measures their abundance as a function of the mass-to-charge ratio (m/ z) (the mass spectrum). From the mass spectrum, conclusions can be drawn as to the structure of the chemical compound.
[0202] As used herein, the term "liquid chromatography-mass spectrometry (LCMS)" denotes an analytical technique that combines the physical separation capability of liquid chromatography with the analytical capability of mass spectrometry. In the liquid chromatography step, the sample is introduced into a column packed with a stationary phase, separating the chemical compounds in the sample by their retention time (Rt) in the column. The chemical compound(s) associated with a retention time interval is then directed to a mass spectrometer, to obtain a mass spectrum that allows conclusions to be drawn as to the structure of this chemical compound(s).
[0203] As used herein, the term "thin layer chromatography (TLC)" denotes an analytical technique that separates chemical compounds in a sample by the different rates at which a plate coated with a stationary phase material is drawn.
[0204] As used herein, the term "Nuclear Magnetic Resonance (NMR) Spectroscopy" denotes an analytical technique that measures the intensity of a resonance response of an array of nuclei to a radio frequency pulse to enable information about the electronic environment to be obtained. of the nuclei. From this, conclusions can be drawn about the chemical structure of the compound in which the nuclei reside. A nuclear magnetic resonance spectroscopy technique that uses hydrogen nuclei (protons) is called proton nuclear magnetic resonance spectroscopy (1HNMR).
[0205] The term "ester" is used herein as is conventional in the field of organic chemistry. For For example, the term "ester" may denote a carbonyl group with attached oxygen and alkyl or an oxygen with attached carbonyl and alkyl.
[0207] As used herein, the term "metabolic syndrome" denotes a biological or medical disorder of energy utilization and storage in an animal or human, which may be characterized by abdominal obesity, elevated blood pressure, elevated fasting plasma glucose, triglycerides elevated serum levels, and/or low levels of high-density cholesterol.
[0209] As used herein, the term "polymerase chain reaction" denotes a biomedical technique for generating many copies of a particular DNA sequence.
[0211] As used herein, the term "milling" denotes a method of purifying a material in which the crude material is washed with a solvent. The solvent can be selected such that the desired product is insoluble and the impurities are soluble, in which case the purified product is left in solid form and the impurities are removed with the solvent. Conversely, the solvent can be selected such that the desired product is soluble and the impurities are insoluble, in which case the purified product is in solution and the impurities are removed as solids. The solvent can be removed, for example by evaporation, to obtain the purified product.
[0213] As used herein, the term "Boc protection" denotes the functionalization of a chemical compound with a tert-butyloxycarbonyl (Boc) group as a protecting group. This allows the chemical compound as a whole to be treated with reagents that would otherwise undesirably attack the unprotected group. The protected group can subsequently be deprotected to produce the desired parent group.
[0215] Exemplary compounds
[0217] The present invention provides a molecule having the structure of a compound of the structure of Formula 1:
[0222] (2S,3S,6R,7aR)-1-(pyridin-2-ylmethyl)-4-(3,3,3-trifluoropropanoyl)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridin-3 -yl pivalate and stereoisomers thereof. This compound can be prepared by the reaction sequences described in the Schemes set forth in Example 1.
[0224] Pharmaceutical Compositions and Administration
[0226] The compounds of the present invention are useful as pharmaceutical agents and can be incorporated into pharmaceutical compositions comprising a therapeutically effective amount of a compound of the invention, as defined herein, and a pharmaceutically acceptable carrier or diluent.
[0228] The compounds of the invention may also be used in the manufacture of derivative compounds which are useful as pharmaceutical agents, and which may likewise be incorporated into pharmaceutical compositions prepared with a therapeutically effective amount of said derivative compound and a pharmaceutically acceptable carrier or diluent.
[0230] The compounds of the invention, and such derivatives thereof, may be useful in the treatment of conditions, diseases and disorders in humans and animals. Such compounds may be formulated as pharmaceutical compositions and administered to a subject in need of treatment, eg, a mammal, such as a human patient, in a variety of forms adapted to the chosen route of administration. For example, the compounds of the invention may be formulated for administration by the oral, nasal, intraperitoneal, or parenterally, intravenously, intramuscularly, topically, or subcutaneously, or by tissue injection.
[0232] Thus, the compounds of the invention may be administered systemically, for example, orally, in combination with a pharmaceutically acceptable carrier such as an inert diluent or assimilable edible carrier, or by inhalation or insufflation. They may be enclosed in soft or hard shell gelatin capsules, may be compressed into tablets, or may be incorporated directly with the foods of the patient's diet. For oral therapeutic administration, the compounds may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, dissolvable pastilles, capsules, elixirs, suspensions, syrups, wafers, and the like. The compounds can be combined with an inert powder carrier and inhaled by the subject or insufflated. Such compositions and preparations must contain at least 0.1% of a compound of the present invention. The percentage of the compound of the invention present in such compositions and preparations may, of course, vary and may conveniently be between about 2% and about 60% by weight of a given unit dosage form. The amount of the compound in such therapeutically useful compositions is such that an effective dosage level will be obtained.
[0234] Tablets, dissolvable tablets, pills, capsules, and the like may also contain the following: binders such as gum tragacanth, acacia, corn starch, or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, fructose, lactose or aspartame, or a flavoring agent such as peppermint, oil of wintergreen or cherry flavor may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as vegetable oil or polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For example, tablets, pills, or capsules can be coated with gelatin, wax, shellac, or sugar, and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propylparabens as preservatives, a dye, and flavorings such as cherry or orange flavor. Of course, any material used in the preparation of any unit dosage form must be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the compounds can be incorporated into sustained release preparations and devices. For example, the compounds can be incorporated into sustained release capsules, sustained release tablets, sustained release pills, and sustained release polymers or nanoparticles.
[0236] The compounds can also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the compounds can be prepared in water, optionally mixed with a non-toxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof, and in oils. Under normal conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms.
[0238] Pharmaceutical dosage forms suitable for injection or infusion may include sterile aqueous solutions or dispersions or sterile powders comprising the compounds which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the final dosage form must be sterile, fluid, and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a liquid solvent or dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, non-toxic glyceryl esters, and mixtures. suitable for them. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions, or by the use of surfactants. Prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases it will be preferable to include isotonic agents, eg sugars, buffers or sodium chloride. Prolonged absorption of injectable compositions can be brought about by the use of absorption delaying agents in the compositions, for example, aluminum monostearate and gelatin.
[0240] Sterile injectable solutions are prepared by incorporating the compounds in the required amount in the appropriate solvent with various of the other ingredients listed above, as required, preferably followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying techniques, which produce a powder of the active ingredient plus any additional desired ingredients present in the previously sterile filtered solutions.
[0242] For topical administration, the compounds can be applied in neat form. However, it may be desirable to administer them to the skin as compositions or formulations, in combination with a dermatologically acceptable carrier, which may be a solid or liquid.
[0244] Useful solid carriers include finely divided solids such as talc, clay, cellulose microcrystalline, silica, alumina and the like. Other solid carriers include non-toxic polymeric nanoparticles or microparticles. Useful liquid carriers include water, alcohols or glycols, or water/alcohol/glycol mixtures, in which the compounds can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants. Adjuncts such as fragrances and additional antimicrobial agents may be added to optimize properties for a given use. The resulting liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using pump or aerosol type sprayers.
[0245] Thickeners such as synthetic polymers, fatty acids, salts and esters of fatty acids, fatty alcohols, modified celluloses, or modified mineral materials may also be used with liquid carriers to form pastes, gels, ointments, soaps, and other spreads for application directly to the skin. user's skin.
[0246] Examples of useful dermatological compositions that can be used to deliver the compounds to the skin are known in the art; for example, see Jacquet et al. (US Patent No. 4,608,392), Geria (US Patent No. 4,992,478), Smith et al. (US Patent No. 4,559,157) and Wortzman (US Patent No. 4,820,508).
[0247] The concentration of the therapeutic compounds of the invention in such formulations can vary widely depending on the nature of the formulation and the intended route of administration. For example, the concentration of the compounds in a liquid composition, such as a lotion, may preferably be about 0.1-25% by weight, or more preferably about 0.5-10% by weight. The concentration in a semi-solid or solid composition, such as a gel or powder, may preferably be about 0.1-5% by weight, or more preferably about 0.5-2.5% by weight.
[0248] Effective dosages and routes of administration of the agents of the invention are conventional. The exact amount (effective dose) of the agent will vary from subject to subject depending on, for example, the subject's species, age, weight and general or clinical condition, the severity or mechanism of any disorder being treated. . particular agent or vehicle used, the method and schedule of administration, and the like. A therapeutically effective dose can be determined empirically, by standard procedures known to those of skill in the art. See, eg, The Pharmacological Basis of Therapeutics, Goodman and Gilman, eds., Macmillan Publishing Co., New York. For example, an effective dose can be initially estimated in cell culture assays or in suitable animal models. The animal model can also be used to determine appropriate concentration ranges and routes of administration. Such information can be used to determine useful doses and routes of administration in humans. Methods for extrapolation of effective doses in mice and other animals to humans are known in the art; for example, see US Patent No. 4,938,949. A therapeutic dose can also be selected by analogy with dosages for comparable therapeutic agents.
[0249] The particular mode of administration and dosage regimen will be selected by the treating physician, taking into account the particular details of the case (eg, the subject, the disease, the disease state involved, and whether the treatment is prophylactic). Treatment may involve daily or multiple daily doses of compounds over a period of a few days to months, or even years.
[0250] In general, however, a suitable dose will be in the range of from about 0.001 to about 100 mg/kg of body weight per day, preferably from about 0.01 to about 100 mg/kg of body weight per day, more preferably from about 0.1 to about 50 mg/kg of body weight per day, or even more preferred, in a range of from about 1 to about 10 mg/kg of body weight per day. For example, a suitable dose may be about 1 mg/kg, 10 mg/kg, or 50 mg/kg of body weight per day.
[0251] The compounds are conveniently administered in unit dosage form; for example, containing from about 0.05 to about 10,000 mg, from about 0.5 to about 10,000 mg, from about 5 to about 1,000 mg, or from about 50 to about 500 mg of active ingredient per unit dosage form.
[0252] Compounds can be administered to achieve peak plasma concentrations of, for example, from about 0.25 to about 200 pM, from about 0.5 to about 75 pM, from about 1 to about 50 pM, from about 2 to about 30 pM. , or from about 5 to about 25 pM. Exemplary desirable plasma concentrations include at least 0.25, 0.5, 1, 5, 10, 25, 50, 75, 100, or 200 pM. For example, plasma levels can be from about 1 to about 100 micromolar or from about 10 to about 25 micromolar. This can be achieved, for example, by intravenous injection of a 0.05 to 5% solution of the compounds, optionally in saline, or administered orally as a bolus containing from about 1 to about 100 mg of the compounds. . Desirable blood levels can be maintained by continuous or intermittent infusion.
[0253] The compounds may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example as one dose per day or as two, three, four or more sub-doses per day. The subdose itself may be further divided, for example, into a number of discrete, closely spaced administrations; as multiple inhalations from an insufflator.
[0255] Example 1: Synthesis of a compound of Formula I
[0257] A compound of Formula 1 was synthesized from the compound of Formula 19 (scopolamine [51-34-3]) ((2S)-(1R,2R,4S,5S)-9-methyl-3-oxa-9 -azatricyclo[3.3.1.02.4]nonan-7-yl-3-hydroxy-2-phenylpropanoate hydrobromide trihydrate) by the steps described below in Schemes 1 to 18.
[0259] A first step is illustrated in Scheme 1.
[0264] Into a 10 liter four neck round bottom flask, sodium borohydride (172 g, 4558 mmol) was added portionwise over about 2 hours to a mechanically stirred suspension of a compound of Formula 19 (333 g, 760 mmol). in 3 liters of absolute ethanol in an ice bath. During this time, gas formation occurred and the suspension was stirred while warming to room temperature overnight. While heating, at about 10°C, sudden additional gassing and foaming occurred.
[0266] The milky suspension was then concentrated to about half its original volume (ie about 3 L to 1.5 L) with an additional precipitate observed, which produced the batch. 5M HCl in isopropyl alcohol (IPA) (5.318 mmol, 1.064 L) was then diluted with 2 L of technical diethyl ether (Et ² Ü). The obtained hydrochloric acid (HCl) solution was then added dropwise to the ice-cold batch, while stirring. The white suspension was allowed to stir mechanically overnight to allow complete hydrolysis of the borate salts.
[0268] The reaction mixture was filtered and the resulting solid was rinsed twice with 500 ml portions of Et ² Ü. The dry solid (containing some Et ² Ü) was dissolved in a minimal amount of 10% aqueous potassium carbonate (K ² CO ³ ) solution (~1.5 L) until a clear solution was obtained. 200 mL of brine and ~50 g of solid NaCl were added to the solution. The aqueous phase was then thoroughly extracted with chloroform/methanol (MeOH)/[7N NH ³ in MeOH] (85:14:1). This procedure was performed 5 times with 1.0 L portions of this solvent mixture each.
[0270] The combined organic extracts were dried (sodium sulfate (Na ² SO ⁴ )), filtered, and the solvent removed under reduced pressure to give 102.2 g (659 mmol) of a compound of Formula 18 ((1R,2R, 4S,5S)-9-methyl-3-oxa-9-azatricyclo[3.3.1.02.4]nonan-7-ol) as a lightly tan oil in 87% yield. ^1H NMR (CDCl ³ ) (Figure 1) showed structural match to the compound of Formula 18 with minor amounts of impurities. ^1H NMR (400 MHz, Chloroform -d) 64.03-4.00 (m, 1H), 3.67 (s, 2H), 3.20-3.18 (m, 2H), 2.52 (s, 3H), 2.14 2.08 (m, 2H), 1.69-1.37 (m, 3H).
[0272] The next step proceeded as illustrated in Scheme 2.
[0275] To a solution of the compound of Formula 18 (102.2 g, 659 mmol), benzoic acid (BzOH) (97 g, 790 mmol), and triphenylphosphine (PPh ³ ) (207 g, 790 mmol) in 1000 mL of dry tetrahydrofuran ( THF) was added a solution of diisopropyl azodicarboxylate (DIAD) (160 g, 790 mmol, 154 mL) in 100 mL dry THF dropwise over a period of 4 hours. During the addition, the solution was kept at -35 to -25°C using acetone/dry ice. The clear colorless solution was removed from the ice bath and stirred at room temperature overnight.
[0277] Samples were taken and analyzed, and the analysis showed that the reaction was complete. The reaction mixture was concentrated, dissolved in 1 L of ethyl acetate (EtOAc), extracted with 1 L of saturated sodium bicarbonate (NaHCO ³ ) and subsequently with 2M aqueous HCl (1x1 L, 2x0.5 L). . The combined acidic aqueous fractions were washed once more with 1 L EtOAc. Approximately 400 g of potassium carbonate (K ² CO ³ ) was added portionwise to the acidic aqueous layer, while stirring, until no further gas formation was observed. The pH of the resulting solution was slightly basic and slightly cloudy and yellow.
[0279] The aqueous phase was then extracted with a 9:1 dichloromethane (DCM)/MeOH solution (3x, 1 L each) and the combined organic fractions dried over sodium sulfate (Na ² SO ⁴ ), filtered and concentrated. to provide 118.3 g (447 mmol) of a compound of Formula 17 ((1R,2R,4S,5S,7r)-9-methyl-3-oxa-9-azatricyclo[3.3.1.02.4]nonan-7 -yl benzoate), which was then confirmed by MS (Figure 2) to be 98% pure with a 67.9% yield. ^1H NMR (400 MHz, Chloroform-d) or 8.07-7.93 (m, 2H), 7.59-7.48 (m, 1H), 7.44 7.40 (m, 2H), 5.39-5.30 (m, 1H), 3.63 (s, 2H), 3.42-3.25 (m, 2H), 2.57 (s, 3H), 2.10-2.04 (m, 2H), 1.92-1.86 (m, 2H).
[0281] The next step continued as illustrated in Scheme 3.
[0286] To a solution of the compound of Formula 17 (201.9 g, 779 mmol) in chloroform (350 mL) under nitrogen atmosphere (not stream), was added K ² CO ³ (452 g, 3270 mmol) and chloroformate of ethyl (279 g, 2569 mmol, 247 mL) to form a light yellow suspension which was then stirred at reflux overnight.
[0288] A sample was then taken and analyzed to show that the reaction had reached 74% conversion to the product, a compound of Formula 16 (1R,2R,4S,5S,7r)-ethyl 7-(benzoyloxy)-3-oxa -9-azatricyclo[3.3.1.02.4]nonane-9-carboxylate). The mixture was further stirred at reflux temperature for another 24 hours.
[0290] Another sample was taken and analyzed which showed that the reaction had reached 75% conversion to product. To drive the reaction to completion, additional K ² CO ³ (53.8 g, 389 mmol) and ethyl chloroformate (85 g, 779 mmol, 74.8 mL) were added to the reaction solution and the mixture was stirred. at reflux temperature overnight.
[0292] After stirring and refluxing overnight, another sample was taken and analyzed to show that the reaction had reached 81% conversion to the compound of Formula 16.
[0293] The reaction mixture was then diluted with 500 mL of DCM and the organic layer was washed with 750 mL of half-saturated aqueous NaHCO3 solution, 750 mL of 0.4 M aqueous HCl, and 750 mL of brine. The mixture was then dried over Na2SO4, then filtered and concentrated under reduced pressure to give a yellow oil. 300 mL of heptane was added and the mixture was vigorously stirred overnight.
[0294] A white suspension had formed containing large white lumps which were ground with a spatula. The suspension was filtered through a glass filter, rinsed with about 250 ml of heptane and about 200 ml of pentane. The suspension was then dried using a vacuum oven for 3 hours to afford the compound of Formula 16 as a white solid (219.6 g, 692 mmol, 89% yield). LCMS of the product showed greater than 95% percent yield, with mass and structure agreement with the desired product as shown by MS (Figure 3B) and 1HNMR (Figure 3A). 1HNMR (400 MHz, Chloroform-d) or 8.01-7.97 (m, 2H), 7.61-7.53 (m, 1H), 7.48-7.42 (m, 2H), 5.48-5.39 (m, 1H), 4.58 (m, 1H), 4.48 (m, 1H), 4.16 (q, J = 7.1 Hz, 2H), 3.56 - 3.53 (m, 2H), 2.34 - 2.21 (m, 2H), 1.98-1.86 (m, 2H), 1.27 (t, J = 7.1 Hz, 3H).
[0295] The next step proceeded as illustrated in Scheme 4.
[0299] In a 6 L three-necked flask, sodium borohydride (157 g, 4152 mmol) was added to a suspension of the compound of Formula 16 (219.6 g, 692 mmol) in 1.5 L of absolute ethanol at room temperature . The reaction was exothermic, having an internal temperature of over 60°C for a period of about 4 hours, extreme gas/foam formation was observed during the reaction. The suspension was magnetically stirred at 50°C overnight.
[0300] A sample was then taken and analyzed by TLC to show that the reaction was complete. The resulting product was a white solid which was stopped on the magnetic stirrer overnight. The mixture was concentrated under reduced pressure and the white solid residue was partitioned between 1 liter of chloroform and 3.5 liters of half-saturated aqueous NaHCO ³ solution. The layers were then separated and the aqueous layer was extracted with additional chloroform (2x, 1 L each). The combined organic layers were washed with 1 L of brine, dried over Na ² SO ⁴ , filtered and concentrated under reduced pressure to give ca. 220 g of the product as a white solid which was stirred in 0.6 L of heptane. overnight with a magnetic stirrer.
[0301] The mixture was then filtered, the product formed spheres which were ground and to which 500 ml of heptane was added. The mixture was vigorously stirred overnight with a magnetic stirrer.
[0302] After stirring the mixture overnight, the whitish suspension still contained spheres which were then crushed with a spatula. The suspension was filtered and the residue was rinsed with ca. 300 mL of heptane and dried in vacuo to give ca. 148 g of product.
[0303] A sample was taken and analyzed by ^1H NMR to show that the structure was consistent with the compound of Formula 15 (1R,2R,4S,5S,7r)-ethyl 7-hydroxy-3-oxa-9-azatricyclo[3.3. 1.02.4] nonane-9-carboxylate), (Figure 4).
[0304] The residue was then stirred in ca. 300 mL Et ² O for 1 hour. The white suspension was filtered; and the residue was rinsed again with ca. 300 mL Et ² O and then dried in vacuo (low N ² flow) to give the compound of Formula 15 (122 g, 572 mmol, 82% yield). ^1H NMR (400 MHz, Chloroform -d) or 4.50 (m, 1H), 4.41 (m, 1H), 4.23-4.09 (m, 3H), 3.42-3.39 (m, 2H), 2.15-2.08 (m, 2H ), 1.73-1.62 (m, 2H), 1.44 (d, J = 5.9 Hz, 1H), 1.26 (t, J = 7.1 Hz, 3H).
[0305] The next step continued as illustrated in Scheme 5.
[0308] Triethylamine (22.78 g, 225 mmol, 31.4 mL) and mesyl-Cl (23.64 g, 206 mmol, 16.08 mL) were added dropwise to a solution of the compound of formula 15 (40 g, 188 mmol) in DCM (500 mL) at 0°C. Once the addition was complete, the ice bath was removed and the slightly milky suspension was stirred while warming to room temperature.
[0310] After 1 hour a sample was taken and analyzed by TLC which showed that complete conversion had occurred. The reaction mixture was then washed twice with 500 ml of water. The DCM layer appeared milky and was dried over Na2SO4 (which made the layer lighter), then filtered and concentrated under reduced pressure to give a thick oil. The oil was stripped twice with toluene to give 54.2 g of light tan solid containing 21 wt% toluene.
[0312] The solid was further dried in vacuo at 50°C until the weight remained constant at 43.2 g (148 mmol; 78.9% yield) yielding a compound of Formula 14 ((1R,2R,4S,5S,7r )-ethyl 7-((methylsulfonyl)oxy)-3-oxa-9-azatricyclo[3.3.1.02.4]nonane-9-carboxylate). A sample was taken and the structure was confirmed by 1HNMR (Figure 5). 1HNMR (400 MHz, Chloroform-d) 65.11-5.02 (m, 1H), 4.54-4.53 (m, 1H), 4.44-4.43 (m, 1H), 4.13 (q, J = 7.1 Hz, 2H), 3.47- 3.45 (m, 2H), 3.00 (s, 3H), 2.28-2.23 (m, 2H), 2.00-1.90 (m, 2H), 1.25 (t, J = 7.1 Hz, 3H).
[0314] The next step proceeded as illustrated in Scheme 6.
[0319] Potassium cyanide (12.14 g, 186 mmol) and 18-crown-6 (1,4,7,10,13,16-hexaoxacyclooctadecane) (0.493 g, 1.864 mmol) were added to a solution of the compound of Formula 14 (19.89 g, 62.1 mmol, 91%) in 300 mL of dry dimethyl sulfoxide to form a pale yellow solution which was stirred at 65°C for two and a half days, or about 65 hours, to produce a solution. light brown
[0321] A sample was taken and analyzed by TLC (heptane/DME 1:1, molybdate staining required), which showed a clean conversion to the desired product (no exoepimeric side product observed). However, at this time, the reaction was found to be incomplete as starting material was also observed. Stirring continued for a total of 118 hours, after which the brown solution was allowed to cool to room temperature, and was combined with an additional batch before being partitioned between 2 L EtOAc and 2 L water.
[0323] The layers were separated and the organic layer was washed twice with 1 L of brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, a compound of Formula 13 ((1R,2R, 4S,5S,7s)-ethyl 7-cyano-3-oxa-9-azatricyclo[3.3.1.02.4]nonane-9-carboxylate). The product was purified by gravity column chromatography (750 g silica, heptane/[5->50% EtOAc]) to give 15.1 g of a white solid, or a compound of Formula 13. A sample was taken and was analyzed by 1HNMR (Figure 6) which showed the product to be consistent with the structure of Formula 13, although the product contained 10% by weight of the exo-side product (which was not problematic for subsequent reactions) and the 7 0.5% by weight of heptane. The combined yield from all experiments was 7.55 g, or 45% yield, after correction for solvent and by-product content. 1HNMR (400 MHz, Chloroform-d) 64.53-4.52 (m, 1H), 4.43-4.41 (m, 1H), 4.12 (q, J = 7.1 Hz, 2H), 3.70-3.68 (m, 2H), 2.93-2.89 (m, 1H), 2.22-2.12 (m, 2H), 2.04- 1.98 (m, 2H), 1.24 (t, J = 7.1 Hz, 3H).
[0325] The next step continued as illustrated in Scheme 7.
[0330] Formula 13 Formula 12.a Formula 12
[0331] Scheme 7
[0333] A 50% Raney nickel in water slurry was added to a solution of the compound of Formula 13 (18.20 g, 82 mmol) in 350 mL MeOH/200 mL ammonia (7N in MeOH). The solution was kept under a nitrogen atmosphere and the Raney nickel slurry was added until a dark black suspension was obtained while stirring vigorously.
[0335] The reaction vessel was evacuated and filled with balloons of H ² , which was repeated twice, and then stirred at 45°C under an atmosphere of H ² created by the balloons. After 3 hours, a sample was taken and analyzed by TLC using heptane/dimethoxyethane (DME) 1:1, which showed that the reaction was complete. The reaction mixture was filtered over a short pad of celite that had been previously rinsed with MeOH. The residue was also rinsed with additional MeOH.
[0337] The filtrate was then concentrated under reduced pressure to give a pale yellow oil. This crude product consisted primarily of the open amines of a compound of Formula 12.a (1R,2R,4S,5S,7s)-ethyl 7-(aminomethyl)-3-oxa-9-azatricyclo[3.3.1.02.4] nano-9-carboxylate and, to a lesser extent, the (desired) cyclized amine a compound of formula 12 (rac-(2R,3S,6S,7aS)-ethyl 3-hydroxyoctahydro-1H-2,6-methanepyrrolo[3, 2-b]pyridine-1-carboxylate).
[0339] To drive the major endoisomer to completion, the intermediate was dissolved in 500 mL absolute ethanol, which created a pale yellow solution, which was then stirred and refluxed overnight.
[0341] A sample was taken, concentrated under reduced pressure, dissolved in CDCl ³ and analyzed by ^1H NMR (FIG. 7) which showed that the intermediate open endoisomer had cyclized. It was further shown that about 9% of the product was open exoamine, with some solvent remaining. ^1H NMR (400 MHz, Chloroform-d) 64.46-4.01 (m, 5H), 3.50-3.44 (m, 1H), 3.16-3.11 (m, 1H), 3.96-2.93 (m, 1H), 2.10-1.66 ( m, 5H), 1.47 (d, J = 13.3 Hz, 1H), 1.26 (t, J = 7.1 Hz, 3H).
[0343] The main batch, a yellow solution, was concentrated under reduced pressure and the residue redissolved in 500 mL CHCl ³ and dried over Na ² SO ⁴ . The solution was filtered and concentrated to give 21.7 g of a compound of Formula 12 as a thick yellow oil containing solvent and the open exoamine used in the next step.
[0344] The next step continued as illustrated in Scheme 8.
[0349] Formula 12 Formula 11
[0351] Scheme 8
[0353] Benzaldehyde (22.74g, 214mmol, 21.72ml) was added to a solution of the compound of formula 12 (37.3g, 165mmol) in 1000ml dichloromethane. After 15 minutes STAB (55.9 g, 264 mmol) was added. The suspension was then stirred at room temperature overnight.
[0355] The reaction mixture was washed with 1 L of water and 1 L of NaHCO3. The organic layer was dried over Na2SO4 and concentrated to dryness to give 55 g of reacted product, which was then purified by gravity column chromatography (600 g, Hep/5-60% ETOAc) to give: 2.2 g of exo-Bn2N adduct; and 35.3 g of a compound of Formula 11 (rac-(2R,3S,6S,7aS)-ethyl 3-hydroxyoctahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1-carboxylate) as was analyzed and confirmed by 1HNMR (FIG. 8B) and MS (FIG. 8A).
[0356] 1HNMR (400 MHz, Chloroform-d) 67.35-7.30 (m, 4H), 7.26-7.22 (m, 2H), 4.41-4.02 (m, 5H), 3.83-3.78 (m, 1H), 3.66 (d, J = 13.3 Hz, 1H), 3.30 - 3.26 (m, 1H), 3.11-3.06 (m, 1H), 2.35-2.31 (m, 1H), 2.07-1.88 (m, 3H), 1.77-1.65 (m, 2H ), 1.44 (d, J = 13.9 Hz, 1H), 1.25 (t, J = 7.1 Hz, 3H).
[0358] The next step continued as illustrated in Scheme 9.
[0362] Scheme
[0364] Imidazole (15.19 g, 223 mmol) and tert-butyldiphenylchlorosilane (30.7 g, 112 mmol, 28.7 mL) were added to a solution of the compound of Formula 11 (35.3 g, 112 mmol) in 100 mL of dry N,N-dimethylformamide to form a pale yellow solution which was stirred at room temperature overnight.
[0366] After stirring was complete, a sample was taken and analyzed by LCMS which showed that the reaction was complete.
[0368] The solution was then concentrated under reduced pressure to give an oily residue which was diluted with 750 mL of DCM and washed with 750 mL of 1:1 saturated aqueous NaHCO3 solution and water. Then the solution was washed with 750 ml of brine. The organic layer was dried over Na2SO4, filtered and concentrated to give ca. 65 g of reacted product as confirmed by TLC.
[0370] The reacted product was purified by gravity column chromatography (ca. 600 g, Hep/5-15% EtOAc) which provided 59.5 g, or 90% yield, of a compound of Formula 10 (rac-(2R, 3R,6S,7aS)-ethyl 4-benzyl-3-((tert-butyldiphenylsilyl)oxy)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1-carboxylate) as very thick colorless oil . A sample was taken and analyzed by 1HNMR (FIG. 9B) and LCMS (FIG. 9A), which showed that the product matched the structure of Formula 10 and contained 6% w/w heptane. 1HNMR (400 MHz, Chloroform-d) 67.72-7.66 (m, 4H), 7.47-7.36 (m, 6H), 7.26-7.16 (m, 3H), 7.12-7.09 (m, 2H), 4.62-4.48 (m , 1H), 4.26 (s, 1H), 4.22-4.03 (m, 3H), 3.40-3.29 (m, 2H), 2.89-2.78 (m, 2H), 1.92-1.76 (m, 4H), 1.62-1.52 (m, 1H), 1.31-1.23 (m, 3H), 1.17-1.11 (m, 1H), 1.02 (s, 9H).
[0375] Iodotrimethylsilane (75.0 g, 375 mmol, 51 mL) was added to a solution of the compound of Formula 10 (73.9 g, 124 mmol, 93%) in 1.2 L of dry toluene to create a yellow reaction mixture. which was stirred at 85°C overnight.
[0377] A sample was then taken and analyzed by TLC, which showed that the reaction was complete. The resulting reaction mixture was a dark solution and was allowed to cool to room temperature (slurry) and quenched with 250 mL MeOH. The mixture was then concentrated to approximately 250 mL. After which 750 mL of DCM was added and the mixture was washed with 750 mL of saturated aqueous NaHCO3/H2O 1:1 solution. The organic layer was then washed with 750 mL of brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to provide about 72 g, or 92% yield, of a compound of Formula 9 (rac-(2R, 3R,6S,7aS)-4-benzyl-3-((tert-butyldiphenylsilyl)oxy)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine) as a dark yellow/orange oil. A sample was taken and analyzed by LCMS (Figure 10) which showed the correct mass, and the product was approximately 80% pure, with the peak at 0.448 being toluene. 1HNMR (400 MHz, Chloroform-d) or 7.69-7.63 (m, 4H), 7.47-7.37 (m, 6H), 7.26-7.12 (m, 5H), 4.36 (s, 1H), 3.73-3.70 (m, 1H), 3.39 (d, J = 13.7 Hz, 1H), 3.26 (d, J = 7.6 Hz, 1H), 3.06 (s, 1H), 2.90 (d, J = 13.7 Hz, 1H), 2.79-2.74 ( m, 1H), 2.41 (bs, 1H), 1.90-1.80 (m, 4H), 1.67-1.64 (m, 1H), 1.11-0.99 (m, 10H).
[0379] The next step proceeded as illustrated in Scheme 11.
[0384] Et ³ N (48.3 g, 477 mmol, 0.067 L) and di-tert-butyl dicarbonate (Boc ² O) (39.1 g, 179 mmol) were added to a solution of the compound of Formula 9 (72 g , 119 mmol, 80%) in 1 L of dichloromethane to form a light yellow solution which was stirred at room temperature over the weekend.
[0386] A sample was taken and analyzed by TLC which showed that the reaction was complete. The solution was diluted with 250 mL of DCM and washed with 1 L of saturated aqueous NaHCO ³ solution and 1 L of brine. The organic layer was dried over Na ² SO ⁴ , filtered, and concentrated to give ca. 80 g of crude product.
[0388] Purification by gravity column chromatography (800 g, heptane/[EtOAc 1 ->10%]) provided 68.4 g, or a 94% yield, of a compound of Formula 8 (rac-(2R,3R, 6S,7aS)-tert-butyl 4-benzyl-3-((tert-butyldiphenylsilyl)oxy)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1-carboxylate) as colorless glass.
[0390] A sample was taken and analyzed by ^1H NMR (Figure 11A) and LCMS (Figure 11B) which showed agreement between the product and the structure of Formula 8, and also showed that the product contained 4% w/w heptane. . ^1H NMR (400 MHz, Chloroform-d) or 7.73-7.65 (m, 4H), 7.47-7.35 (m, 6H), 7.24-7.10 (m, 5H), 4.53-4.40 (m, 1H), 4.24 (d, J = 3.8 Hz, 1H), 4.10-3.92 (m, 1H), 3.44-3.32 (m, 2H), 2.87 (d, J = 13.6 Hz, 1H), 2.33- 2.77 (m, 1H), 1.93-1.72 (m, 4H), 1.65-1.54 (m, 1H), 1.50-1.47 (m, 9H), 1.10-1.02 (m, 10H).
[0392] The next step continued as illustrated in Scheme 12.
[0396] Under a flow of nitrogen, palladium, 10% on activated carbon (7 g, 125 mmol) was added to a solution of the compound of Formula 8 (72.9 g, 125 mmol) in 600 mL of acetic acid. The container was closed and the resulting mixture was stirred at 50°C for 2 hours under an atmosphere of hydrogen created by a balloon.
[0398] The mixture was then stirred at 50°C overnight. The black suspension was filtered over celite rinsed with EtOH, and the filtrate was concentrated under reduced pressure. The residue was separated twice with 0.5 L of toluene, after which it was dissolved in 1 L of diethyl ether.
[0400] The organic layer was then washed with 1 L of 10% (w/v) aqueous K ² CO ³ solution, 1 L of brine, dried over Na ² SO ⁴ , filtered and concentrated under reduced pressure before being separated again with pentane to give 58.5 g of a thick tan syrup, a compound of Formula 7 (rac-(2R,3S,6S,7aS)-tert-butyl 3-((tert-butyldiphenylsilyl)oxy) octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1-carboxylate).
[0402] A sample was taken and analyzed by ^1H NMR (Figure 12B) and LCMS (Figure 12A) which showed the product to be consistent with the structure of Formula 7 and to contain 5.1 wt% toluene and 1.3 wt% by weight of n-pentane. ^1H NMR (400 MHz, Chloroform-d) or 7.68-7.63 (m, 4H), 7.45-7.35 (m, 6H), 4.40-4.25 (m, 1H), 4.13-3.93 (m, 2H), 3.41-3.36 (m, 1H), 2.97-2.92 (m, 1H), 2.62 (d, J = 11.5 Hz, 1H), 1.96-1.78 (m, 2H), 1.67 (s, 1H), 1.64-1.56 (m, 1H ), 1.49-1.47 (m, 9H), 1.16-1.13 (m, 1H), 1.05-1.04 (m, 9H).
[0404] The compound of Formula 7 was separated into its respective enantiomers by supercritical fluid chromatography (SFC) on a Welkho-1 column with 90/10 scCO ² /iPrOH 0.2% isopropylamine eluent as illustrated in Scheme 13.
[0407] The next step continued as illustrated in Scheme 14.
[0412] 3,3,3-Trifluoropropanoic acid (3.629 mL, 41.1 mmol, 1.5 eq) was dissolved in DCM (120 mL) and dry DMF (10 mL). DIPEA (7.16 mL, 41.1 mmol, 1.5 eq) and HATU (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3 -oxydhexafluorophosphate) (15.63 g, 41.1 mmol, 1.5 eq) and the mixture was stirred at room temperature for 1.5 hours. This resulted in the formation of a clear, reddish-brown solution.
[0414] To that solution, a solution of the compound of Formula 6.b (13.5 g, 27.4 mmol) in DCM (100 mL) was added and the solution was stirred at room temperature for 4 hours.
[0416] The reaction mixture was diluted with DCM (250 mL), washed with 1M aqueous KHSO4 (400 mL), saturated aqueous NaHCO3 (400 mL), water (400 mL), brine (250 mL), dried over Na2SO4 and concentrated in vacuo to give 22.74 g (>100%) of a compound of Formula 5 ((2S*,3S*,3aS*,6R*,7aR*)-tert-butyl 3-((tert-butyldiphenylsilyl)oxy) -4-(3,3,3-trifluoropropanoyl)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1-carboxylate) as a brown oil.
[0418] The next step continued as illustrated in Scheme 15.
[0423] The compound of Formula 5 (max 27.4 mmol) was dissolved in dry THF (115 mL).
[0425] A solution of tetrabutylammonium fluoride in THF (1M, 82 mL, 82 mmol) was added and the reaction mixture was stirred at 50°C overnight. LCMS analysis revealed complete conversion to the desired material.
[0427] The solution was concentrated in vacuo and coevaporated twice with 50% EtOAc/heptane (2x, 100 mL each) to give 38.66 g of crude material as a brown oil. The material was dissolved in 25% EtOAc/Et ² O (800 mL) and washed with water (2*600 mL each). The aqueous layers were combined and extracted with 25% EtOAc/Et ² O (400 mL). The organic layers were combined, washed with brine (400 mL), dried over Na ² SO ⁴ and concentrated in vacuo to give 15.14 g of material as a brown oil.
[0429] Purification by gravity column chromatography (gradient 50% EtOAc/heptane to 100% EtOAc) yielded 5.85 g of a compound of Formula 4 ((2S*,3S*,3aS*,6R*,7aR*) -tert-butyl 3-hydroxy-4-(3,3,3-trifluoropropanoyl)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1-carboxylate) (58% in 2 steps) as a white foam.
[0430] The next step proceeded as illustrated in Scheme 16,
[0434] The compound of Formula 4 (5.85 g, 16 mmol) was dissolved in pyridine (50 mL), followed by the addition of DMAP (dimethylaminopyridine) (1.96 g, 16.06 mmol) and pivaloyl chloride (3, 95ml, 32.1mmol).
[0436] The reaction mixture was stirred overnight at 60°C. LCMS analysis revealed complete conversion to the desired material. The reaction mixture was allowed to cool to room temperature (a light brown suspension formed) and concentrated in vacuo.
[0438] The residue was diluted with EtOAc (250 mL) and washed with 0.5M aqueous KHSO ⁴ solution (200 mL) and saturated aqueous NaHCÜ ³ (250 mL). Each time the aqueous layer was extracted with additional EtOAc (50 mL).
[0440] The combined organic layers were washed with brine (200 ml), dried over sodium sulfate, filtered and evaporated to dryness to give 6.8 g of crude material. Purification by flash column chromatography (EtOAc/heptane gradient) provided 5.49 g (76%) of a compound of Formula 3 ((2S*,3S*,3aS*,6R*,7aR*)-tert-butyl 3 -(pivaloyloxy)-4-(3,3,3-trifluoropropanoyl)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridine-1-carboxylate) as a white foam. LCMS analysis: purity > 95%, found 449.3 [M+H]+ and 393.2 (M-(C4H8)+H]+).
[0442] The next step continued as illustrated in Scheme 17.
[0447] The compound of Formula 3 (1g, 2.23mmol) was dissolved in DCM (20ml).
[0449] TFA (trifluoroacetic acid) (8.54 mL, 111 mmol) was added and the mixture was stirred at room temperature for 1 h. LCMS analysis revealed complete conversion to the desired material.
[0451] The reaction mixture was concentrated in vacuo and coevaporated with toluene (2x, 20 mL each). The residue was dissolved in chloroform (40 mL) and washed with saturated aqueous Na ² CO ³ solution (40 mL). The aqueous phase was extracted with chloroform (3x, 20 mL each).
[0453] The organic layers were combined, washed with brine (70 mL), dried (Na ² SO ⁴ ), filtered and evaporated under reduced pressure to give 769.9 mg (99%) of a compound of Formula 2 (( 2S*,3S*,3aS*,6R*,7aR*)-4-(3,3,3-trifluoropropanoyl)octahydro-1H-2,6-methanepyrrolo[3,2-b]pyridin-3-ylpivalate) as an off-white solid. The structure was confirmed by ^1H NMR as shown in Figure 13.
[0454] The next step proceeded as illustrated in Scheme 18,
[0458] The compound of Formula 2 (693.8mg, 1.99mmol) was dissolved in DCM (20ml).
[0459] 2-Pyridinecarboxaldehyde (284 mL, 2.99 mmol) was added and the mixture was stirred for 2 hours. Sodium triacetoxyborohydride (696 mg, 3.29 mmol) was then added and the mixture was stirred overnight at room temperature.
[0460] LCMS analysis revealed complete conversion to the desired material. The mixture was concentrated in vacuo. The residue was dissolved in chloroform (40 mL) and washed with saturated aqueous Na2C03 solution (40 mL). The aqueous phase was extracted with chloroform (3x, 20 mL each). The organic layers were combined, washed with brine (70 mL), dried (Na2SO4), filtered and evaporated under reduced pressure to give crude material.
[0461] Purification by column chromatography (silica, gradient 14% EtOAc/heptane to 100% EtOAc) provided 718 mg of oily material. The oil was dissolved in EtOAc (~10 mL) and filtered through a paper filter, again concentrated in vacuo and successively co-evaporated with acetonitrile, pentane and Et ² O to give 658 mg (75%) of a compound of Formula 1 ((2S*,3S*,3aS*,6R*,7aR*)-1-(pyridin-2-ylmethyl)-4-(3,3,3-trifluoropropanoyl)octahydro-1H-2,6- methanepyrrolo[3,2-b]pyridin-3-yl pivalate) as a foam. The structure was confirmed by ^1H NMR analysis as shown in Figure 14, and the absolute stereochemistry was confirmed by X-ray analysis.
[0462] A general description of these synthetic steps to transform the starting reagent into a compound of Formula 1 is provided in Scheme 19, below.
[0463]
[0466] The Taxol model was used to investigate the MoA of Formula 1.
[0468] Animals showing chemotherapy-induced pain after repeated doses of taxol were treated with either vehicle gabapentin or Formula 1. Plasma was collected after the first dose of the test articles and after 7 days of daily treatment. The skin of the hind paws was also collected after 7 days of daily treatment. Tissues were processed and expression levels of 1310 proteins were evaluated (Somascan provided by SomaLogic, Inc., Boulder, CO).
[0469] Three statistical methods were used to identify protein expression that discriminated between Formula 1, gabapentin, and vehicle on study day 8, after a single dose, and study day 14, after multiple dosing.
[0471] A comparison between day 8 and day 14 within each group was also made. All methods were performed with scaled data. First, a Student's t-test was performed to compare proteins between groups. Second, the LASSO method (less absolute selection and contraction operator) was applied. This is a regression technique that simultaneously performs the selection of variables and the modification of the coefficient of the model. These models used cluster as the outcome and all 1310 proteins as initial predictors. The LASSO was then adjusted to reduce the unimportant coefficients to zero, leaving approximately the ten most important predictors (or discriminators between) the groups.
[0473] The final method used was Random Forest. The Random Forest is an ensemble method that randomly selects subsets of proteins, creating classification trees based on the subset to discriminate between the two groups. The process was repeated 2000 times. The varying importance of each protein is ranked based on the yield of the proteins in the trees.
[0475] The LASSO method automatically provides the top 10 predictors of group discrimination. T-test and Random Forest results were ordered by p-value and mean precision decline, respectively, and the top 10 of these results were selected. Additional barriers were placed on the top 10 predictors selected by statistical analysis to screen for more significant changes: (1) a selected protein must rank among the top 10 major proteins in plasma and skin; (2) the change must be in the same direction in the plasma and in the skin; and (3) the p-value in the Student's t-test must be equal to or less than 0.01.
[0477] This screening and selection process revealed a single protein that fits all the criteria, 14-3-3a (STRATIFIN gene). Biostatistical analysis showed that in skin, 14-3-3a ranked second using the LASSO classification method and 8 using the Random Forest classification method. Student's t-test versus vehicle showed a p-value of 0.0110. Exploring changes in protein obtained in plasma when comparing day 8 and day 14, 14-3-3a ranked 7 using the LASSO ranking method, and Student's t-test p-value was <0.01 .
[0479] Figure 15 shows the relative decrease in stratifin in percentages from day 8 to day 14 after treatment with vehicle, gabapentin, or Formula 1. Treatment with Formula 1 resulted in a nearly 50% decrease in stratifin vs. animals. untreated. These changes were significant versus the changes observed in vehicle-treated animals. Formula 1 treatment significantly reduced 14-3-3a expression in the skin at study day 14 versus vehicle-treated animals (FIG. 16).
[0481] Example 3: Putative MoA via 14-3-3o Targeting: Biological Significance
[0483] According to Li et al. discovered that selected 14-3-3 isomers, including the o-isomer, interact with the Ca ^v 2.2 ion channel, a subtype of voltage-gated Ca ²⁺ channels that is central to neurotransmitter release at the presynaptic nerve terminal . They reported that protein 14-3-3 (E, t, y, or y isomers)^ reduced Ca ²⁺ channel inactivation in both the open and closed states. This protein-protein interaction is mediated by the binding of 14-3-3 or the carboxyl tail of the pore-forming α1B subunit of the channel that contains at least two potential 14-3-3 interaction sites.
[0485] Cav2.2 is an N-type calcium channel. This channel mediates neurotransmission of pain signals in the spinal cord, making its suppression a desirable target for the treatment of chronic pain. Prialt® (Jazz Pharmaceuticals, ziconotide), a selective Cav2.2 blocker, has been approved by the FDA for the treatment of chronic pain. Although it has been shown to be effective in treating some patients with chronic pain, the drug has serious limitations, including its invasive (intrathecal) route of administration and its short therapeutic window.
[0487] Unlike the currently available Cav2.2 ziconotide blocker for pain treatment, Formula 1 may offer some advantages. Formula 1 does not directly target CaV2.2, but indirectly modulates by reducing 14-3-3, leading to a reduction in 14-3-3/Cav2.2 interaction and ultimately a reduction in Ca ²⁺ influx. Unlike the reported side effects of ziconotide, no side effects or tolerability effects were observed after treatment with Formula 1. A cumulative analgesic response was observed in some of the neuropathic pain models and a more durable response was observed in other models. Formula 1 is the only Cav2.2 modulator active by oral administration.
[0488] Example 4: Senerga® Phenotypic Screening Data
[0489] Formula 1 was discovered after extensive phenotypic selection of Senerga®. The compound showed activity in pain-related models, without affecting the withdrawal threshold of untreated animals, suggesting a limited CNS effect. Formula 1 was not effective on swelling, thus excluding an NSAID-like mechanism of action.
[0490] The compound demonstrated a good pharmacological safety profile, with no immediate alarm events (no effect on blood pressure, no post-mortem findings, no effect on body weight or food intake, and no effect on specific CNS-related tests). These initial data suggest that Formula 1 and its chemical analogs may be candidates for the treatment of pain through a potentially novel mechanism of action.
[0491] Example 5: In vitro absorption of bioavailability
[0492] 5.1: Permeability using monolayers of Caco-2 cells
[0493] Formula 1 was evaluated for cell permeability using monolayers of Caco-2 cells. Both apical-to-basolateral (A-B) and basolateral-to-apical (B-A) bidirectional patency and efflux ratios were evaluated in the trial. The observed Papp A-B was 6.6x10-6 cm/s, demonstrating moderate passive permeability (10x10-6 cm/s Papp AB 1x10-6 cm/s is considered moderate permeability).
[0494] The Papp B-A observed was high, at 68.3x10-6 cm/s, leading to an efflux ratio of 10.4. This high efflux ratio suggested that the compound is a substrate for one or more efflux transporters. When an efflux transporter inhibitor (Pgp inhibitor) was added in the assay, the PappA-B value increased from 6.6x10-6 cm/s to 45.2x10-6 cm/s, showing that the compound is a good substrate for this efflux transporter (Pgp)
[0495] 5.2: Physicochemical properties
[0496] Formula 1 has good physicochemical properties (MW<500Da, cLogP and cLogD=2.6, and no "Rule of Five" violations). The low polar surface area (PSA<70Á2) also suggests that the compound should be a good membrane and brain penetrant (Table 1).
[0497] Table 1 : Physicochemical properties of Formula 1
[0499]
[0501] Example 6: Pharmacokinetic Studies
[0502] Pharmacokinetic studies of MD-354 were performed in mice after oral administration of 50 mg/kg and intravenous administration of 5 mg/kg (FIG. 17). After intravenous administration of 5 mg/kg, the initial plasma concentration (C0) reached 7941 ng/ml. Plasma concentration decreased in a multiphasic manner to 6.74 ng/mL at 8 hours post-dose, with a terminal half-life of 0.35 hours (excluding data at the 8-hour time point). The compound demonstrated a moderate systemic plasma clearance (CLp) of 46.1 mL/min/kg (mouse hepatic blood flow rate: 90 mL/min/kg) and a moderate steady-state volume of distribution (Vss) of 1.08 L. /kg.
[0503] After oral administration of 50 mg/kg, a high maximum plasma concentration (Cmax) was reached within 5 minutes, reaching 1933 ng/ml. Plasma concentration decreased in a multiphasic manner to 0.76 ng/mL at 8 hours post-dose. The terminal half-life was 0.85 h. The absolute oral bioavailability was relatively 10.5%.
[0504] Two additional sets of oral data in mice showed similar pharmacokinetic profiles in all studies. Peak 354 plasma concentrations were high, at 3157 ng/mL and 3001 ng/mL, all achieved within 5 minutes. Plasma concentrations decreased to 8.4 ng/ml and 11.4 ng/ml at 8 hours after administration, with a terminal half-life of 1.19 h and 1.25 h, respectively. Both data sets showed oral bioavailability values of 14.1% and 14.7%, respectively.
[0505] Example 7: Tissue Distribution
[0506] 7.1 In Vitro Studies: Plasma Protein Binding
[0507] Plasma protein binding assay for Formula 1 was performed in mouse and human plasma at a single concentration of 1 | ^j M using the equilibrium dialysis method. The compound showed moderate to high plasma protein binding with 84% and 92% plasma protein bound in human and mouse plasma, respectively. Plasma protein binding of the compound is acceptable.
[0508] 7.2 In vivo studies
[0509] 7.2.1 Formula 1 levels in mouse tissues
[0510] Tissue distribution studies were performed by administering the compound to mice at a dose of 50 mg/kg orally and intravenously. Plasma and tissue samples were collected at various time points from 5 min to 8 h. The tissue-to-plasma concentration ratios measured after oral dosing were: 0.6 (range: 0.3-1, 1, brain), 3.2 (range: 1.9-5.0, spinal cord) , 6.5 (range: 0.9-10.3, liver), 24.0 (range: 5.1-59.9, lung), 6.6 (range: 1.2-14.9, heart ), 16.9 (range: 4.3-26.8, kidney), 77.5 (range: 11.5-341, spleen), 29.2 (range: 6.3-50.2, cecum), 67.4 (range: 11.6-326, colon), 5.2 (range: 2.1-8.0, testis). The concentration-time profiles of the compound in these tissues generally followed those in plasma. Tissue-to-plasma concentration ratios in these selected tissues are generally within the normal ranges for marketed drugs.
[0511] In general, a substantial effort is needed to discover CNS drugs that penetrate the brain and spinal cord. Brain levels are commonly used as a surrogate for spinal cord levels when the test compound is not mediated by uptake or efflux transporters. The efflux transporter Pgp is highly expressed on the luminal side of the blood-brain barrier and blood spinal cord barrier. When a drug is a substrate for Pgp, such as Formula 1, the compound must have a higher concentration in the CSF than in the brain, since the compound is pumped out of the brain at the BBB but into the CSF at the BCSFB, which makes the compound available to enter the spinal cord tissue.
[0512] There are complex equilibria between the concentrations of compounds in the blood, CSF, and spinal cord. A high concentration in the CSF would produce a large concentration in the spinal cord. As Formula 1 is a substrate of Pgp and is highly tissue bound, its higher concentrations in spinal cord tissue compared to brain tissue are not surprising (Table 2).
[0513]
[0514] Example 8: Metabolism Transporter
[0515] 8.1 In vitro metabolism
[0516] 8.1.1 Stability in blood/plasma
[0517] Formula 1 was stable in both mouse and human plasma, with t90 values greater than 4 h, while it was unstable in rat plasma, with t90 less than 1 h (t90 should be greater than 4 h to be considered stable). Therefore, most of the efficacy and safety pharmacology has been performed in mice.
[0518] 8.1.2 Metabolic Stability in Liver Microsomes/Hepatocytes Between Species
[0519] Metabolic stability studies were performed on mouse, rat, and human liver microsomes at a compound concentration of 1 pM. Intrinsic clearance (CLint) values were >346 μl/min/mg in both mouse and rat liver microsomes (t1/2 of <4 min) and 174 μl/min/mg in human liver microsomes (t1/ 2 of 8 min). This may indicate that the compound undergoes significant pre-systemic metabolism or a first-pass effect in the liver of the species tested, since the compound shows good cell permeability.
[0520] The results of this study showed that N-dealkylation and oxidation appeared to be the main metabolic pathways. A total of five metabolites were detected in the three species. There were no significant differences in metabolic profiles between mouse and rat liver microsomes, suggesting that either mouse or rat can be used as rodent species for GLP toxicology studies. Due to the similarity in plasma stability of the compound in mouse and human plasma, toxicology studies would preferably be performed in the mouse. No unique human metabolites were detected in this study.
[0521] 8.2 Enzyme inhibition and induction
[0522] 8.2.1 Inhibition of CYP450 enzymes
[0523] Preliminary in vitro studies on reversible inhibition of major CYP450 enzymes were performed in a cocktail format for this compound. IC50 values were 4 pM for CYP2C9, 5 pM for CYP2C19, and greater than 10 pM for CYP3A4, 2C8, 2D6, and 1A2. If total plasma concentrations in humans are less than 0.4 pM, then in vivo drug interaction studies in clinical trials may not be required for this compound.
[0524] Example 9: Excretion
[0525] 9.1 In vivo studies
[0526] The fecal excretion of the parent compound was evaluated after oral administration of Formula 1 in mice at a dose of 50 mg/kg and intravenous administration at a dose of 5 mg/kg. Peak concentrations in feces were 150 pg/g and 33 pg/g for oral and intravenous dosing, respectively. These peak concentrations were generally achieved within 1 hour after oral administration and within 5 minutes after intravenous administration. Concentrations remained high (greater than 1 pg/g) even after 8 h after dosing by both routes of administration (FIG. 18).
[0527] Example 10: Non-clinical safety pharmacology
[0528] 10.1 In Vitro Safety Pharmacology Profiles
[0529] The IC50 for hERG was greater than 10 pM. A generally accepted safety margin against hERG and other important cardiac targets is >30 (a safety margin is calculated as: IC50/Cmax, free or EC50/Cmax, free). This suggests that a total plasma concentration of at least 4 pM can be achieved in humans with a safety margin of 30.
[0530] Screening for Cerep activity was performed and was negative against selected targets at 10 pM (including some important cardiovascular safety related targets) for Formula 1.
[0531] 10.2 In Vivo Safety Pharmacology Profiles
[0532] 10.2.1. General safety remarks
[0533] The tolerability of Formula 1 was evaluated in mice and rats. The compound was well tolerated in mice after 5 days of intraperitoneal dosing at 30 mg/kg, subcutaneous dosing at 60 mg/kg, and oral dosing at 50 mg/kg orally. The compound was also tolerated after a single dose of 100 mg/kg in the mouse. At rats dosed orally, once daily with Formula 1 at 50 mg/kg for a period of 5 days, showed no adverse effects (Table 3).
[0534] Table 3: Safety pharmacology data after repeated dosing of Formula 1 using if r n vi mini r i n. NPF - in h ll z li .
[0536]
[0538] 10.3 CNS Safety Pharmacology
[0539] to. open field test
[0540] Mice received an IP dose of 10 mg/kg of Formula 1. Animals were placed in the open field apparatus after 30 minutes (box 0.5 X 0.5 X 0.5 m in length, width and height). ) for a period of 5 minutes. The walking distance and speed of the animals were evaluated and compared with the vehicle-treated group. No differences were found between animals treated with vehicle and animals treated with Formula 1 (Table 4).
[0541] Table 4
[0545] b. food consumption
[0546] The animals were tested for animal feed consumption over a period of 3 consecutive days during which they received a daily dose of Formula 1 at 10 mg/kg IP. No changes were detected in food consumption of animals treated with Formula 1 versus animals treated with vehicle. Vehicle-treated animals consumed a mean of 4.39 ± 0.61 g per night compared to 4.00 ± 0.20 g per night for Formula 1-treated animals.
[0547] c. Body weight
[0548] Formula 1 dosing to untreated mice and untreated rats over a period of 31 days and 7 days, respectively, did not affect body weight gain. At 4 days, vehicle-treated animals gained approximately 1% body weight from 25.24±0.74 g to 25.50±0.64 g. Animals treated with Formula 1 went from 25.28±1.04 g to 25.82±1.25 g.
[0549] Animals treated with Formula 1 at a dose of 10 mg/kg IP over a period of 31 days gained 15.64% body weight (changing from 21.44±0.18 g to 25.49±0.46 g). . Vehicle-treated animals gained 18.9% body weight (changing from 21.45 ± 0.19 g to 23.97 ± 0.62 g; p = 0.07), therefore, no significant changes were found.
[0550] d. Response of untreated animals to hot and cold stimuli.
[0551] Changes in normal withdrawal threshold responses to hot and cold stimuli in untreated animals after pharmacological treatment may reflect an effect on the CNS, as in the case of cannabinoids or opioids. Therefore, the response of untreated mice after administration of Formula 1 at a dose of 10 mg/kg IP was evaluated (Table 5). Formula 1 was not effective in altering the basal response of animals stimulated with cold (2±1°C) or heat (50±1°C), indicating no CNS effect. This may also suggest that Formula 1 is acting on a target that is elevated or exposed in pain state and is less relevant in the untreated state.
[0552] Table 5: Latency of response of untreated animals to hot-cold stimuli
[0554]
[0556] Example 11: In vivo activity of Formula 1 in models of neuropathic pain
[0557] The IP route of administration was used for proof of principle for initial studies with Formula 1. However, the route of administration of choice in most studies conducted with Formula 1 was SC or PO. A brief summary of the protocol is provided in each section, for the purpose of understanding the results without going through all the detailed procedures.
[0558] Data are presented as mean±SD; the number of animals per group is presented in the study outline table; p<0.05 was considered a significant change. The efficacy summary table also includes the activity of the analogs.
[0559] 11.1 Activity of SC Administered Formula 1 on Taxol-Induced Neuropathic Pain in Mice
[0560] Brief Summary of Study: Animals were dosed daily with Taxol for a period of 8 consecutive days (Day 0 to Day 7). On day 8 of the study, before drug dosing, the decrease in withdrawal force (tactile allodynia) was verified by applying the von Frey test. After assigning the animals to their treatment groups, the animals were treated daily for a period of 5 days with vehicle or Formula 1 at different doses. Gabapentin at a dose of 150 mg/kg was dosed only on test days (day 8 and day 14).
[0561] Data presented in Figure 19 shows that 8 days after Taxol dosing, vehicle-treated animals experienced significant low withdrawal strength. This low withdrawal strength was maintained throughout the study period. Gabapentin treatment resulted in increased withdrawal strength over a 4-hour period. indicating analgesia activity. Formula 1 treatment at a dose as low as 1 mg/kg resulted in profound analgesic activity over a period of 24 h to 3 days.
[0562] Response to heat stimuli was assessed using the hot plate apparatus set at 50°C. Daily Taxol treatment resulted in a decrease in latency response time. At baseline prior to Taxol dosing, the animals' response time to heat was 28.10 ± 1.60 seconds (vehicle-treated group). Eight days after Taxol treatment, the response time was significantly reduced (9.90±2.18 seconds, p<0.01). On study days 13 and 17, the response time was significantly shorter than at baseline (11.70±2.11 and 11.10±1.79 seconds, respectively). Treatment with Formula 1 at doses of 10 mg/kg, 3 mg/kg and 1 mg/kg was active in increasing response time. However, it is interesting to note that the beneficial activity of Formula 1 on heat hyperalgesia was more marked on study day 14, after repeated daily dosing of the compound, than on study day 8 after a single dose ( Table 6). It is suggested that the activity of Formula 1 increases over time, rather than decrease as would be expected due to tolerance with currently used drugs.
[0563] Table 6: The effect of Formula 1 on thermal hyperalgesia (heat, 50°C)
[0565]
[0567] 11.2 Activity of Formula 1 in a Mouse Chronic Constriction Injury (CCI) Model Using the SC Route of Administration
[0568] Brief summary of the study: The sciatic nerve of mice was exposed under anesthesia and 3 loose ligatures were placed around the nerve. Animals were allowed to recover for 3 days. Tactile allodynia (von Frey test) and thermal hyperalgesia (heat at 50±1°C and cold at 2±1°C) were then assessed. Animals experiencing tactile allodynia were assigned to their treatment groups (vehicle, gabapentin, and Formula 1 at different doses). Subsequently, the animals were treated daily for a period of 5 days with vehicle or Formula 1 at different doses. Gabapentin at a dose of 150 mg/kg was dosed only on test days (day 3 and day 7).
[0569] Three days after CCI, vehicle-treated animals experienced a reduction in withdrawal strength after the von Frey test. The low strength of withdrawal was also shown on day 7 of the study (FIG. 20). Gabapentin treatment at a dose of 150 mg/kg resulted in a significant, but not complete, reversal in withdrawal strength. Treatment with Formula 1 at a dose of 60 mg/kg resulted in a significant increase in withdrawal strength after repeated daily treatment.
[0570] In addition to decreased withdrawal strength, vehicle-treated animals showed increased sensitivity to cold, as observed by the reduction in the animals' response time to a cold plate (2YC) (FIG. 21). Gabapentin treatment at a dose of 150 mg/kg increased the response time of the animals to values above the reference value before the operation, suggesting an adverse effect on the CNS. Treatment with Formula 1 at 30 mg/kg and 60 mg/kg resulted in increased response time to cold stimuli. Activity peaked at 2 h after a single dose (study day 4) and at 2 h after dosing on study day 7 after repeated daily dosing (study day 6; Figure 21). .
[0571] No tolerance effect was observed for Formula 1 after repeated dosing; on the contrary, repeated dosing led to an increase in the activity of the compound. The effect of Formula 1 on the response time in the hot plate test was not significant.
[0572] 11.3 Formula 1 is active using the oral route of administration
[0573] The activity of Formula 1 after oral administration was evaluated using the Taxol model (see Study Scheme in section 7.1). Formula 1 at a dose of 50 mg/kg was active in completely reversing the withdrawal force at 2 h after single administration on study day 8. This activity was maintained at study day 14 after dosing. repeated daily from day 8 to day 14. Interestingly, Formula 1 was significantly active at 24 h post-dosing with increasing strength of withdrawal (study day 14, pre-dosing, Figure 22). No tolerance effect was observed. Gabapentin dosed at 150 mg/kg was active for a period of 4 h after dosing. No activity was found when the animals were placed in the hot plate test.
[0574] In this study, the effect of Formula 1 on heat hyperalgesia was not statistically different from the vehicle-treated group.
[0575] 11.4 Summary of Activity in Mice
[0576] Table 7 summarizes the activity of Formula 1 in mouse models of neuropathic pain after repeated daily administration using different routes of administration. The following is observed: (1) Formula 1 treatment at relatively low doses was as good as gabapentin at 150 mg/kg, and in some cases even better than gabapentin; (2) The duration of activity of Formula 1 is t and longer than gabapentin; (3) Gabapentin treatment at a dose of 150 mg/kg resulted in an increase in response time to cold beyond the initial response time, suggesting a clear CNS effect. Treatment with Formula 1 shows activity without CNS effect.
[0577] T l 7: R m n livi 4 n m l nim l r l l r n r i n r n .
[0579]
[0581] 11.5 Formula 1 Is Orally Active in Taxol-Induced Neuropathic Pain in the Rat
[0582] Brief summary of the study: Taxol was administered daily for a period of 17 days (day 0 to day 16). On day 16, the strength of withdrawal was assessed using the von Fey test. The animals experienced a greater than 60% reduction in withdrawal force after repeated administration of Taxol. After a single treatment with Formula 1 at a dose of 50 mg/kg, a complete reversal was recorded, ie, the strength of withdrawal was similar to the values recorded before the start of Taxol dosing. The duration of analgesic activity was at least 6 h. There were no significant differences between the vehicle and Formula-treated groups 124 h after dosing (pre-dosing day 23). The activity of Formula 1 was similar to the activity observed after treatment with gabapentin at a dose of 150 mg/kg (FIG. 23).
[0583] Multiple doses of Taxol resulted in a reduction in body weight that continued throughout the study period and did not cease even after Taxol administration was discontinued on day 15. In Formula 1 treatment, the rate of weight loss was reduced (Figure 24), suggesting that Formula 1 improved the general welfare of the animals.
[0584] 11.6 Orally Administered Formula 1 Is Active in CCI-Induced Neuropathic Pain in the Rat
[0585] Brief Study Plan: Male SD rats were operated on as described by Bennet and Xie7. On study day 7, the animals were tested for sensitivity to von Frey. Only animals that showed a significant reduction in withdrawal strength were assigned to the treatment groups. Animals were then dosed with vehicle, gabapentin, or Formula 1. Formula 1 treatment was ineffective 2 h after the single dose. However, after daily treatment with Formula 1 at a dose of 50 mg/kg, the activity of the compound increased and on study day 11 there was a significant increase in the withdrawal threshold at 2 h after treatment, which suggests pain relief (Figure 25).
[0586] The embodiments illustrated and discussed in this specification are intended only to teach those skilled in the art the best way known to the inventors to make and use the invention. Nothing in this specification should be considered as limiting the scope of the present invention. all examples presented are representative and not limiting. The embodiments of the invention described above may be modified or varied, without departing from the invention, as will be appreciated by those skilled in the art in light of the above teachings. Therefore, it is to be understood that, within the scope of the claims, the invention may be practiced other than as specifically described.
[0588] It should be understood that although the compounds of Formulas 1-18 may be drawn with specific chirality for simplicity, one of ordinary skill in the art will recognize how to create and separate these various isomers. Accordingly, all isomers of the compounds of Formulas 1-18 can be understood to be within the scope of the present application.

权利要求:
Claims (23)
[1]
1. A compound of Formula 1:

[2]
2. A pharmaceutical composition comprising a compound of Claim 1, and a pharmaceutically acceptable carrier or diluent.
[3]
3. A compound of Claim 1 for use in a method of treating a subject (human or animal) suffering from a condition, disease or disorder, said method comprising administering to the subject an effective amount of said compound;
optionally wherein the condition or disorder is neuropathic pain or chronic pain.
[4]
4. The compound for use according to claim 3, wherein the compound is administered to effect:
a) localized administration to the subject; or
b) systemic administration to the subject.
[5]
5. A method of making a compound of Formula 1:

[6]
6. The method of claim 5, further comprising making the compound of Formula 2 by reacting a compound of Formula 3:

[7]
7. The method of claim 6, further comprising making the compound of Formula 3 by reacting a compound of Formula 4:

[8]
8. The method of claim 7, further comprising making the compound of Formula 4 by reacting a compound of Formula 5:

[9]
9. The method of claim 8, further comprising making the compound of Formula 5 by reacting a compound of Formula 6.b:

[10]
10. The method of claim 9, further comprising making the compound of Formula 6.b by chirally separating a compound of Formula 7:

[11]
11. The method of claim 10, further comprising making the compound of Formula 7 by reacting a compound of Formula 8:

[12]
12. The method of claim 11, further comprising making the compound of Formula 8 by reacting a compound of Formula 9:

[13]
13. The method of claim 12, further comprising making the compound of Formula 9 by reacting a compound of Formula 10:

[14]
14. The method of claim 13, further comprising making the compound of Formula 10 by reacting a compound of Formula 11:

[15]
15. The method of claim 14, further comprising making the compound of Formula 11 by reacting a compound of Formula 12:

[16]
16. The method of claim 15, further comprising making the compound of Formula 11 by cyclizing a compound of Formula 12.a:

[17]
17. The method of claim 16, further comprising making the compound of Formula 12.a by reacting a compound of Formula 13:

[18]
18. The method of claim 17, further comprising making the compound of Formula 13 by reacting a compound of Formula 14:

[19]
19. The method of claim 18, further comprising making the compound of Formula 14 by reacting a compound of Formula 15:

[20]
20. The method of claim 19, further comprising making the compound of Formula 15 by reacting a compound of Formula 16:

[21]
21. The method of claim 20, further comprising making the compound of Formula 16 by reacting a compound of Formula 17:

[22]
22. The method of claim 21, further comprising making the compound of Formula 17 by reacting a compound of Formula 18:

[23]
23. The method of claim 22, further comprising making the compound of Formula 18 by reacting a compound of Formula 19:

with a reducing agent;
optionally wherein the reducing agent is sodium borohydride;
optionally wherein the reaction further comprises HCl in isopropyl alcohol.

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法律状态:

优先权:

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申请号 | 申请日 | 专利标题

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US201562214734P| true| 2015-09-04|2015-09-04|

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US201562214727P| true| 2015-09-04|2015-09-04|

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PCT/US2016/049871|WO2017040764A1|2015-09-04|2016-09-01|Therapeutic compounds for pain and synthesis thereof|

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