Pharmaceutical compositions of 6-(2-(2H-tetrazol-5-yl)ethyl)-6-fluorodecahydroisoquinoline-3-carboxylic acid and its ester derivatives.

Abstract

6-(2-(2H-tetrazol-5-yl)ethyl)-6-fluorodecahydroisoquinoline-3-carboxylic acid and 6-(2-(2H-tetrazol-5-yl)ethyl)-6-fluorodecahydroisoquinoline-3-carboxylic acid hydrocarbyl ester derivatives represented by the formula are disclosed, as are pharmaceutical compositions and methods for treating pain, epilepsy, convulsions, and epileptic seizures using those compositions.

Description

[Technical field]

[0001] The present invention relates to pharmaceutical compositions of 6-(2-(2H-tetrazol-5-yl)ethyl)-6-fluorodecahydroisoquinoline-3-carboxylic acid and 6-(2-(2H-tetrazol-5-yl)ethyl)-6-fluorodecahydroisoquinoline-3-carboxylic acid hydrocarbyl ester derivatives for use in the treatment of pain and epilepsy. [Background technology]

[0002] Alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (also known as AMPA receptors, AMPARs, or quisqualate receptors) are ionotropic transmembrane receptors known as glutamate-gated ion channels that mediate fast synaptic transmission in the central nervous system (CNS). AMPARs have traditionally been classified as non-NMDA-type receptors, along with kainate receptors.

[0003] Glutamate is the major excitatory amino acid transmitter in the central nervous system. AMPA receptors (AMPARs) are large multisubunit ion channels made up of four AMPAR subunit proteins, GluA1, GluA2, GluA3, and GluA4 (encoded by four separate genes). AMPARs are present at excitatory synapses of neurons and are responsible for fast excitatory neurotransmission. AMPARs transmit glutamate signals to depolarize the postsynaptic neuron. Glutamate released from excitatory neurons diffuses across the synapse and binds to AMPARs on the postsynaptic neuron. AMPARs physically penetrate the cell membrane of neurons and contain ion pores or channels that selectively allow the inward flow of primarily sodium ions (but also potassium and, rarely, calcium) from the outside into the cell. In the absence of glutamate, the AMPAR ion pores are closed, and thus ions cannot flow into the neuron. Glutamate binding to AMPARs opens them, allowing primarily sodium ions to pass through the pore across the cell membrane of the postsynaptic neuron, resulting in depolarization. Thus, sodium conducts a depolarizing current. AMPARs are crucial for neuronal networks and the physiology of the brain and central nervous system. Briefly, AMPA receptors are neurotransmitter-gated (glutamate-gated) ion channels that open only in response to the chemical signal glutamate.

[0004] AMPA receptors play an important role in the initiation and spread of epileptic seizures (Scharfman HE.. "The Neurobiology of Epilepsy", Curr Neurol Neurosci Rep., (Germany), 2007, Vol. 7, p.348-354; Rogawski MA., "Revisiting AMPA receptors as an antiepileptic drug target", Epilepsy Currents, (USA), 2011, Vol. 11, p.56-63.). Neurosurgeons and neurologists have observed that in clinical samples taken from microdialysis fluids of the brains (hippocampi) of human epilepsy patients (n=6 patients), glutamate levels increase before and during seizures (During MJ and Spencer DD, “Extracellular Hippocampal Glutamate and Spontaneous Seizure in the Conscious Human Brain”, The Lancet., (Netherlands), 1993, Vol. 341, No. 8861, p.1607-10).

[0005] Treatment with various AMPAR antagonists in animal model studies of seizures, convulsions and epilepsy has consistently demonstrated the preclinical efficacy of this class of compounds, regardless of whether the AMPAR antagonist molecules have a non-competitive or competitive mechanism of action. In early clinical trials, neurologists found that treatment with the experimental therapeutic AMPAR antagonist talampanel reduced seizures in epilepsy patients (Chappell AS, Sander JW, Brodie MJ, Chadwick D, Lledo A, Zhang D, Bjerke J, Kiesler GM, Arroyo S., “A Crossover, Add-On Trial of Talampanel in Patients With Refractory Partial Seizures”, Neurology, (Netherlands), 2002, Vol. 58, p.1680-1682).Ten years later, the AMPAR antagonist perampanel was approved as the first FDA- and EMEA-approved AMPAR antagonist for the treatment of epilepsy (French JA, Krauss GL, Steinhoff BJ, Squillacote D, Yang H, Kumar D, Laurenza A., “Evaluation of adjunctive perampanel in patients with refractory partial-onset seizures: Results of randomized global phase III study 305”, Epilepsia, (USA), 2013, Vol. 54, p.117-125; and Krauss GL, Perucca E, Ben-Menachem E, Kwan P, Shih JJ, Squillacote D, Yang H, Gee M, Zhu J, Laurenza A., “Perampanel, a selective, noncompetitive α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor antagonist, as a adjunctive therapy for refractory partial-onset seizures: interim results from phase III, extension study 307”, Epilepsia, (US), 2013, Vol. 54, p.126-134).

[0006] AMPAR permeability to calcium and other cations, such as sodium and potassium, is regulated by the GluA2 subunit. If an AMPAR lacks the GluA2 subunit, it will be permeable to sodium, potassium, and calcium. The presence of the GluA2 subunit renders the channel almost always impermeable to calcium. This is determined by post-transcriptional modification (RNA editing) of the Q→R editing site in the GluA2 mRNA. Here, A→I (adenosine→inosine) editing (by double-stranded RNA-specific adenosine deaminase (Adenosine Deaminase Acting on RNA) 2) changes the GluA2 RNA that codes for the uncharged amino acid glutamine (Q) in the receptor's ion channel to code for the positively charged arginine (R). The presence of a positively charged amino acid at a critical site makes it energetically unfavorable for calcium to pass through the pore and into the cell. Sodium is the main ion that can permeate the glutamate-gated ion channel of AMPAR.

[0007] Loss of function of double-stranded RNA-specific adenosine deaminase (ADAR), which edits GluA2 mRNA, leads to certain neurological disorders due to the resulting calcium permeability of altered AMPARs. Animals lacking ADAR2 die from seizures by postnatal day 21. However, in epilepsy, normal edited AMPAR receptors are present and are permeable to sodium.

[0008] AMPAR antagonists can reduce AMPAR-mediated overactivation of neuronal networks in epilepsy, and therefore represent potential targets for the treatment of epilepsy. A review article is Rogawski MA., "Revisiting AMPA receptors as an antiepileptic drug target", Epilepsy Currents, (USA), 2011, Vol. 11, P.56-63.

[0009] U.S. Patent No. 5,670,516 discloses that certain decahydroisoquinoline derivatives are AMPA receptor antagonists and are therefore useful for treating various neurological conditions, including pain, migraine, convulsions, and epileptic seizures. In addition, WO 01 / 02367 search report published on January 11, 2001 discloses diester-type prodrug forms of 3S,4aR,6S,8aR-6-(((4-carboxy)phenyl)methyl)-1,2,3,4,4a,5,6,7,8,8a-decahydroisoquinoline-3-carboxylic acid, which is a selective GluR5 antagonist.

[0010] U.S. Pat. No. 7,247,644 discloses that monoesters of the monoacid (3S,4aR,6R,8aR)-6-[2-(1(2)H-tetrazol-5-yl)ethyl-1,2,3,4,4a,5,6,7,8,8a-decahydroisoquinoline-3-carboxylic acid provide significantly improved bioavailability of the monoacid compared to that obtained by administration of the monoacid itself.

[0011] However, the bioavailability of these aforementioned methods is still not high enough to be considered for oral application. Summary of the Invention

[0012] In a first aspect, the present invention provides a compound of formula I: [ka] (In the formula, R is H and (C1-C 20 ) hydrocarbyl The present invention relates to a compound of the formula:

[0013] In a second aspect, the present invention provides a compound of formula II: [ka] The present invention relates to a compound of the formula:

[0014] In a third aspect, compounds of formula II (wherein R=H) are shown to selectively inhibit (S)-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (s-AMPA)-induced AMPAR calcium permeability in vitro and prevent seizures in animal models of epilepsy in vivo.

[0015] In a fourth aspect, the present invention relates to a method or medicament for treating epilepsy and / or pain by administering a compound of formula II as an orally administrable prodrug (R≠H) of the active pharmaceutical ingredient (R=H).

[0016] In a fourth aspect, the present invention relates to a pharmaceutical composition comprising a pharma- ceutically acceptable carrier and a compound as described herein.

[0017] These and other objects, features and advantages of the present invention will become apparent from the following detailed description of the various aspects of the invention. [Brief description of the drawings]

[0018] [Figure 1] FIG. 1 shows the results of s-AMPA induced currents in an in vitro electrophysiological study of pyramidal neurons from Sprague-Dawley rat cerebral cortical slices using compound #1, 6-(2-(2H-tetrazol-5-yl)ethyl)-6-fluorodecahydroisoquinoline-3-carboxylic acid (R=H). [Diagram 2] FIG. 2. Results of s-AMPA induced currents in in vitro electrophysiological studies of pyramidal neurons from Sprague-Dawley rat cerebral cortical slices using compound #1. [Diagram 3] FIG. 1 shows the results of NMDA-induced currents in an in vitro electrophysiology study of pyramidal neurons from Sprague-Dawley rat cerebral cortical slices using compound #1. [Figure 4]FIG. 1 shows the results of a single acute dose pharmacokinetic study of treatment of Sprague-Dawley rats with Compound 1 or Compound 4. Compound 1 was measured by LCMS in plasma samples. [Diagram 5] FIG. 1 shows the results of a single acute dose pharmacokinetic study of treatment of Sprague-Dawley rats with Compound 1 or Compound 4. Compound 1 was measured by LCMS in brain samples. [Figure 6] FIG. 1 shows the results of a single acute dose pharmacokinetic study of treatment of Sprague-Dawley rats with Compound 1 or Compound 4. Compound 1 was measured by LCMS in cerebrospinal fluid (CSF) samples. [Figure 7] 7A-7C show the results of a single ascending dose (SAD) study of acute oral administration of compound 4 as a 1-day treatment in vivo in Sprague-Dawley rats, with compound 1 measured by LCMS in plasma (7A), CSF (7B), and brain (7C) samples at 75 and 300 minutes. [Figure 8] 8A and 8B show the results of an open field test (OFA) on the same animals as in the SAD test of Figure 7. Total distance traveled (8A) and exploration rearing latency (8B) were measured. [Figure 9] 9A-9C show further results from the OFA of Figure 8. Locomotor activity (9A) and stereotypic activity (9B) were counted, and immobility time (9C) was measured. [Figure 10] Figure 10 shows the results of the Irwin neurological test battery for the same animals as in the SAD test of Figure 7. Cage observations (10A), autonomic responses (10B), and the suspension test (10C) were scored, and fecal pellet counts (10D) were performed. [Figure 11] FIG. 1 shows the results of body weight measurements in Sprague-Dawley rats undergoing a multiple ascending dose (MAD) study of once daily dosing for 5 days. [Figure 12] OFA results for the same Sprague-Dawley rats as in Figure 11. Total distance traveled and exploratory rearing time were measured, and ambulatory and stereotypic movements were counted. [Figure 13]FIG. 13 shows the results of the Irwin neurological test battery for the same Sprague-Dawley rats as in FIG. 11. Cage observations (13A), anatomical responses (13B), and the suspension test (13C) were scored, and fecal pellets (13D) were counted (performed on day 3 of the MAD test). [Figure 14] FIG. 16 shows the results of the Irwin neurological test battery for the same Sprague-Dawley rats as in FIG. 13 (performed on day 5 of the MAD test). [Figure 15] 15A-15B show the results of a MAD study in Sprague-Dawley rats administered orally once daily for 5 days. Plasma levels of Compound 1 were measured by LCMS on days 1, 3, and 5; Compound 1 concentrations are plotted against time (15A) and dose (15B). [Figure 16] FIG. 16 shows the results of the same MAD study as in FIG. 15, where levels of Compound 1 in CSF on day 5 were measured by LCMS. [Figure 17] FIG. 15 shows the results of the same MAD study in FIG. 15, where the levels of Compound 1 in the brain on day 5 were measured. Table 1 shows some of the compounds. Table 2 shows the results of a comparison between s-AMPA and NMDA in an in vitro electrophysiology study of pyramidal neurons using 6-(2-(2H-tetrazol-5-yl)ethyl)-6-fluorodecahydroisoquinoline-3-carboxylic acid (R=H). Table 3 shows the results of a study on the in vivo efficacy of 6-(2-(2H-tetrazol-5-yl)ethyl)-6-fluorodecahydroisoquinoline-3-carboxylic acid (R=H).

[0019] [Detailed Description] The present disclosure relates generally to 6-(2-(2H-tetrazol-5-yl)ethyl)-6-fluorodecahydroisoquinoline-3-carboxylic acid and hydrocarbyl ester derivatives of 6-(2-(2H-tetrazol-5-yl)ethyl)-6-fluorodecahydroisoquinoline-3-carboxylic acid, and pharmaceutical compositions thereof, as well as methods of treating certain disorders.

[0020] Throughout the specification the definitions of terms and substituents are retained. For convenience and clarity, certain terms used in the specification, examples, and claims are explained here.

[0021] The terms "hydrocarbyl," "aliphatic hydrocarbyl," "aromatic hydrocarbyl," and "alkyl," as used herein and as described below, include all possible structural features of the defined groups, such as straight-chain, branched, cyclic, polycyclic, bridged, etc.

[0022] "Hydrocarbyl" (or "hydrocarbon") refers to any group composed solely of hydrogen and carbon as elemental components.

[0023] The first subset of hydrocarbyls is "aliphatic hydrocarbyls," which refer to hydrocarbyl groups that are not aromatic. Aliphatic hydrocarbyls are any of a variety of sp hydrocarbyls that are not arranged to be aromatic, as would be readily understood by one of ordinary skill in chemistry. 3 -,sp 2 The aliphatic hydrocarbyl group contains one or more alkanes (sp 3 ), alkenes (sp 2 ), alkynes (sp), and allenes (sp 2 and sp) functional groups. Two or more alkene, alkyne, and / or arene functional groups may be conjugated in a hydrocarbyl group and the group is still defined as an aliphatic hydrocarbyl group so long as the conjugation does not result in aromaticity.

[0024] Examples of aliphatic hydrocarbyl groups include methyl, ethyl, isopropyl, isobutyl, cyclopropyl, t-butyl, neopentyl, 3-methylbutyl, 3,3-dimethylbutyl, 2-propylpentyl, 2-butylhexyl, 2-pentylheptyl, 2-hexyloctyl, n-hexyl, n-octyl, 2-ethylbutyl, 1-methyl-2-ethylbutyl, decyl, dodecyl, tetradecyl, 9-hexadeca-en-yl, 9-octadec-en-yl, 9,12-octadeca-dien-yl, cyclohexyl, cyclohexylmethyl, 2-cyclohexylethyl, dicyclohexylmethyl, 2-butenyl, 2-butynyl, cyclopentyl, norbornyl, and the like.

[0025] A second subset of hydrocarbyl is "aromatic hydrocarbyl" and "aryl", which refers to a hydrocarbyl group that is aromatic. Aromatic hydrocarbyl groups include, for example, phenyl (C6H5), naphthyl (C 10 H7), anthracene, etc. (C 14 H9) is one example.

[0026] The third subset of hydrocarbyls is "alkyl" (or alkane), which is a sp 3 Refers to a hydrocarbyl group consisting only of hybridized carbon atoms. Alkyl groups are also a subset of aliphatic hydrocarbyl groups, except for sp 2It is a fully saturated hydrocarbyl group excluding the presence of - and sp hybridized carbon atoms. Examples of alkyl groups from the above-mentioned examples of aliphatic hydrocarbyl groups include methyl, ethyl, isopropyl, isobutyl, cyclopropyl, t-butyl, neopentyl, 3-methylbutyl, 3,3-dimethylbutyl, 2-propylpentyl, 2-butylhexyl, 2-pentylheptyl, 2-hexyloctyl, n-hexyl, n-octyl, 2-ethylbutyl, 1-methyl-2-ethylbutyl, decyl, dodecyl, tetradecyl, cyclohexyl, cyclohexylmethyl, 2-cyclohexylethyl, dicyclohexylmethyl, cyclopentyl, norbornyl, etc. The alkyl groups do not include, for example, 9-hexadeca-en-yl, 9-octadec-en-yl, 9,12-octadeca-dien-yl, 2-butenyl, 2-butynyl, etc., which are also examples of aliphatic hydrocarbyl groups mentioned above.

[0027] The hydrocarbyl group may be exclusively aliphatic hydrocarbyl, aromatic hydrocarbyl or alkyl in nature. Alternatively, the term hydrocarbyl includes a combination of one or more of the above subsets of groups, typically as a substituent. Thus, an aliphatic hydrocarbyl that is optionally substituted, for example with a phenyl group, can be included in the term "hydrocarbyl" or "aliphatic hydrocarbyl optionally substituted with phenyl". "Alkyl optionally substituted with phenyl" refers to an alkyl group in which all carbon atoms, except for the phenyl group, are sp 3 This indicates that a hybrid orbital is formed.

[0028] As defined above, aliphatic hydrocarbyl, aromatic hydrocarbyl (i.e., aryl), and alkyl, either alone or in combination, are appropriate limitations of hydrocarbyl, provided that the number of carbon atoms is the same or fewer than the number of carbon atoms in the original hydrocarbyl. Similarly, alkyl is a further limitation of aliphatic hydrocarbyl, provided that the number of carbon atoms is the same or fewer than the number of carbon atoms in the original hydrocarbyl.

[0029] As used herein, the term "optionally substituted" may be used interchangeably with "unsubstituted or substituted." The term "substituted" refers to the replacement of one or more hydrogen atoms in a particular group with a specified radical. For example, substituted alkyl, aryl, cycloalkyl, etc. refers to alkyl, aryl, or cycloalkyl in which one or more H atoms in each residue are replaced with a specified substituent.

[0030] As set forth above, the definitions of R for the hydrocarbyl, aliphatic hydrocarbyl, aromatic hydrocarbyl, and alkyl groups are further defined as follows: x -C y ), where x is the minimum number of carbon atoms and y is the maximum number of carbon atoms. For example, "(C1-C 20 "(C5-C)hydrocarbyl" refers to a hydrocarbyl group having 1 to 20 carbon atoms. 14 "Aliphatic hydrocarbyl" refers to an aliphatic hydrocarbyl group having 5 to 14 carbon atoms. Any substituent containing carbon, such as phenyl, may have any number of carbon atoms, e.g., (C x -C y ) is separate from the specification.

[0031] Unless otherwise specified, the term "carbocycle" is intended to include ring systems in which the ring atoms are all carbon, but in any oxidation state. Thus, (C3-C 10 ) Carbocycle refers to both non-aromatic and aromatic systems, including such systems as cyclopropane, benzene, and cyclohexene; (C8-C 12 ) Carbopolycycle refers to systems such as norbornane, decalin, indane and naphthalene. Carbocycle refers to monocyclic, bicyclic and polycyclic rings, unless otherwise limited.

[0032] The compounds described herein may contain double bonds in the substituent R and may further contain other centers of geometric asymmetry; unless otherwise specified, it is intended that the compounds include both E / Z geometric isomers. Similarly, all tautomeric forms, such as the tautomer of tetrazole: [ka] is also intended to be included.

[0033] The compounds may further contain one or more asymmetric centers in the substituent R and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined in terms of absolute stereochemistry as (R)- or (S)-. The present invention is intended to include all such possible isomers, as well as their racemic and optically pure isomers. Optically active (R)- and (S)-isomers may be prepared using chiral synthons or chiral reagents or resolved using conventional techniques.

[0034] For clarity, the atom numbering convention for the decahydroisoquinoline ring system and its adducts is shown below: [ka]

[0035] In various embodiments of the present invention, R is selected from the group consisting of (C1-C 20 ) hydrocarbyl, (C-C 18 ) hydrocarbyl, (C-C 19 ) hydrocarbyl, (C-C 18 ) hydrocarbyl, (C-C 17 ) hydrocarbyl, (C-C 16 ) hydrocarbyl, (C-C 15 ) hydrocarbyl, (C-C 14 ) hydrocarbyl, (C-C 13 ) hydrocarbyl, (C-C 12 ) hydrocarbyl, (C-C 11 ) hydrocarbyl, (C-C 10)hydrocarbyl, (C1-C9)hydrocarbyl, (C1-C8)hydrocarbyl, (C1-C7)hydrocarbyl, (C1-C6)hydrocarbyl, (C1-C5)hydrocarbyl, (C1-C4)hydrocarbyl, and (C1-C3)hydrocarbyl.

[0036] In various embodiments of the present invention, R is (C-C 20 ) hydrocarbyl, (C-C 19 ) hydrocarbyl, (C-C 18 ) hydrocarbyl, (C-C 17 ) hydrocarbyl, (C-C 16 ) hydrocarbyl, (C-C 15 ) hydrocarbyl, (C-C 14 ) hydrocarbyl, (C-C 13 ) hydrocarbyl, (C-C 12 ) hydrocarbyl, (C-C 11 ) hydrocarbyl, (C-C 10 )hydrocarbyl, (C2-C9)hydrocarbyl, (C2-C8)hydrocarbyl, (C2-C7)hydrocarbyl, (C2-C6)hydrocarbyl, (C2-C5)hydrocarbyl, (C2-C4)hydrocarbyl, and (C2-C3)hydrocarbyl.

[0037] In various embodiments of the present invention, R is (C3-C 20 ) hydrocarbyl, (C3-C 19 ) hydrocarbyl, (C3-C 18 ) hydrocarbyl, (C3-C 17 ) hydrocarbyl, (C3-C 16 ) hydrocarbyl, (C3-C 15 ) hydrocarbyl, (C3-C 14 ) hydrocarbyl, (C3-C 13 ) hydrocarbyl, (C3-C 12 ) hydrocarbyl, (C3-C 11 ) hydrocarbyl, (C3-C 10)hydrocarbyl, (C3-C9)hydrocarbyl, (C3-C8)hydrocarbyl, (C3-C7)hydrocarbyl, (C3-C6)hydrocarbyl, (C3-C5)hydrocarbyl, and (C3-C4)hydrocarbyl.

[0038] In various embodiments of the present invention, R is (C4-C 20 ) hydrocarbyl, (C4-C 19 ) hydrocarbyl, (C4-C 18 ) hydrocarbyl, (C4-C 17 ) hydrocarbyl, (C4-C 16 ) hydrocarbyl, (C4-C 15 ) hydrocarbyl, (C4-C 14 ) hydrocarbyl, (C4-C 13 ) hydrocarbyl, (C4-C 12 ) hydrocarbyl, (C4-C 11 ) hydrocarbyl, (C4-C 10 ) hydrocarbyl, (C4-C9) hydrocarbyl, (C4-C8) hydrocarbyl, (C4-C7) hydrocarbyl, (C4-C6) hydrocarbyl, and (C4-C5) hydrocarbyl.

[0039] In various embodiments of the present invention, R is (C5-C 20 ) hydrocarbyl, (C5-C 19 ) hydrocarbyl, (C5-C 18 ) hydrocarbyl, (C5-C 17 ) hydrocarbyl, (C5-C 16 ) hydrocarbyl, (C5-C 15 ) hydrocarbyl, (C5-C 14 ) hydrocarbyl, (C5-C 13 ) hydrocarbyl, (C5-C 12 ) hydrocarbyl, (C5-C 11 ) hydrocarbyl, (C5-C 10 ) hydrocarbyl, (C5-C9) hydrocarbyl, (C5-C8) hydrocarbyl, (C5-C7) hydrocarbyl, and (C5-C6) hydrocarbyl.

[0040] In various embodiments of the present invention, R is (C6-C 20 ) hydrocarbyl, (C6-C 19 ) hydrocarbyl, (C6-C 18 ) hydrocarbyl, (C6-C 17 ) hydrocarbyl, (C6-C 16 ) hydrocarbyl, (C6-C 15 ) hydrocarbyl, (C6-C 14 ) hydrocarbyl, (C6-C 13 ) hydrocarbyl, (C6-C 12 ) hydrocarbyl, (C6-C 11 ) hydrocarbyl, (C6-C 10 ) hydrocarbyl, (C6-C9) hydrocarbyl, (C6-C8) hydrocarbyl, and (C6-C7) hydrocarbyl.

[0041] In various embodiments of the present invention, R is (C7-C 20 ) hydrocarbyl, (C7-C 19 ) hydrocarbyl, (C7-C 18 ) hydrocarbyl, (C7-C 17 ) hydrocarbyl, (C7-C 16 ) hydrocarbyl, (C7-C 15 ) hydrocarbyl, (C7-C 14 ) hydrocarbyl, (C7-C 13 ) hydrocarbyl, (C7-C 12 ) hydrocarbyl, (C7-C 11 ) hydrocarbyl, (C7-C 10 )hydrocarbyl, (C7-C9)hydrocarbyl, and (C7-C8)hydrocarbyl.

[0042] In various embodiments of the present invention, R is (C-C 20 ) hydrocarbyl, (C8-C 19 ) hydrocarbyl, (C8-C 18 ) hydrocarbyl, (C8-C 17 ) hydrocarbyl, (C8-C 16 ) hydrocarbyl, (C8-C 15 ) hydrocarbyl, (C8-C14 ) hydrocarbyl, (C8-C 13 ) hydrocarbyl, (C8-C 12 ) hydrocarbyl, (C8-C 11 ) hydrocarbyl, (C8-C 10 ) hydrocarbyl, and (C8-C9) hydrocarbyl.

[0043] In various embodiments of the present invention, R is (C9-C 20 ) hydrocarbyl, (C9-C 19 ) hydrocarbyl, (C9-C 18 ) hydrocarbyl, (C9-C 17 ) hydrocarbyl, (C9-C 16 ) hydrocarbyl, (C9-C 15 ) hydrocarbyl, (C9-C 14 ) hydrocarbyl, (C9-C 13 ) hydrocarbyl, (C9-C 12 ) hydrocarbyl, (C9-C 11 ) hydrocarbyl, and (C9-C 10 ) hydrocarbyl.

[0044] In various embodiments of the present invention, R is 10 -C 20 ) hydrocarbyl, (C 10 -C 19 ) hydrocarbyl, (C 10 -C 18 ) hydrocarbyl, (C 10 -C 17 ) hydrocarbyl, (C 10 -C 16 ) hydrocarbyl, (C 10 -C 15 ) hydrocarbyl, (C 10 -C 14 ) hydrocarbyl, (C 10 -C 13 ) hydrocarbyl, (C 10 -C 12 ) hydrocarbyl, and (C 10 -C 11 ) hydrocarbyl.

[0045] In various embodiments of the present invention, R is 11 -C 20 ) hydrocarbyl, (C 11 -C 19 ) hydrocarbyl, (C 11 -C 18 ) hydrocarbyl, (C 11 -C 17 ) hydrocarbyl, (C 11 -C 16 ) hydrocarbyl, (C 11 -C 15 ) hydrocarbyl, (C 11 -C 14 ) hydrocarbyl, (C 11 -C 13 ) hydrocarbyl, and (C 11 -C 12 ) hydrocarbyl.

[0046] In various embodiments of the present invention, R is 12 -C 20 ) hydrocarbyl, (C 12 -C 19 ) hydrocarbyl, (C 12 -C 18 ) hydrocarbyl, (C 12 -C 17 ) hydrocarbyl, (C 12 -C 16 ) hydrocarbyl, (C 12 -C 15 ) hydrocarbyl, (C 12 -C 14 ) hydrocarbyl, and (C 12 -C 13 ) hydrocarbyl.

[0047] In various embodiments of the present invention, R is 13 -C 20 ) hydrocarbyl, (C 13 -C 19 ) hydrocarbyl, (C 13 -C 18 ) hydrocarbyl, (C 13 -C 17 ) hydrocarbyl, (C 13 -C16 ) hydrocarbyl, (C 13 -C 15 ) hydrocarbyl, and (C 13 -C 14 ) hydrocarbyl.

[0048] In various embodiments of the present invention, R is 14 -C 20 ) hydrocarbyl, (C 14 -C 19 ) hydrocarbyl, (C 14 -C 18 ) hydrocarbyl, (C 14 -C 17 ) hydrocarbyl, (C 14 -C 16 ) hydrocarbyl, and (C 14 -C 15 ) hydrocarbyl.

[0049] In various embodiments of the present invention, R is 15 -C 20 ) hydrocarbyl, (C 15 -C 19 ) hydrocarbyl, (C 15 -C 18 ) hydrocarbyl, (C 15 -C 17 ) hydrocarbyl, and (C 15 -C 16 ) hydrocarbyl.

[0050] In various embodiments of the present invention, R is 16 -C 20 ) hydrocarbyl, (C 16 -C 19 ) hydrocarbyl, (C 16 -C 18 ) hydrocarbyl, and (C 16 -C 17 ) hydrocarbyl.

[0051] In various embodiments of the present invention, R is 17 -C 20 ) hydrocarbyl, (C17 -C 19 ) hydrocarbyl, and (C 17 -C 18 ) hydrocarbyl.

[0052] In various embodiments of the present invention, R is (C 18 -C 20 ) hydrocarbyl or (C 18 -C 19 ) hydrocarbyl.

[0053] In various embodiments of the present invention, R is (C 19 -C 20 ) hydrocarbyl.

[0054] In various embodiments of the present invention, R is 20 ) hydrocarbyl, (C 19 ) hydrocarbyl, (C 18 ) hydrocarbyl, (C 17 ) hydrocarbyl, (C 16 ) hydrocarbyl, (C 15 ) hydrocarbyl, (C 14 ) hydrocarbyl, (C 13 ) hydrocarbyl, (C 12 ) hydrocarbyl, (C 11 ) hydrocarbyl, (C 10 ) hydrocarbyl, (C9) hydrocarbyl, (C8) hydrocarbyl, (C7) hydrocarbyl, (C6) hydrocarbyl, (C5) hydrocarbyl, (C4) hydrocarbyl, and (C3) hydrocarbyl.

[0055] In some embodiments of the present invention, R is an aliphatic (C1-C2) optionally substituted with 1 or 2 phenyl groups, with the proviso that R contains 20 or fewer carbons. 20 ) hydrocarbyl, aliphatic (C1-C2) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 19 ) hydrocarbyl, aliphatic (C1-C2) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 18) hydrocarbyl, aliphatic (C1-C2) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 17 ) hydrocarbyl, aliphatic (C1-C2) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 16 ) hydrocarbyl, aliphatic (C1-C2) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 15 ) hydrocarbyl, aliphatic (C1-C2) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 14 ) hydrocarbyl, aliphatic (C1-C2) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 13 ) hydrocarbyl, aliphatic (C1-C2) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 12 ) hydrocarbyl, aliphatic (C1-C2) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 11 ) hydrocarbyl, aliphatic (C1-C2) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 10 ) hydrocarbyl, aliphatic (C1-C9) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, aliphatic (C1-C8) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, aliphatic (C1-C7) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, aliphatic (C1-C6) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, aliphatic (C1-C5) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, aliphatic (C1-C4) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, and aliphatic (C1-C3) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons.

[0056] In various embodiments of the present invention, R is an aliphatic (C2-C20 ) hydrocarbyl, aliphatic (C2-C3) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 19 ) hydrocarbyl, aliphatic (C2-C3) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 18 ) hydrocarbyl, aliphatic (C2-C3) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 17 ) hydrocarbyl, aliphatic (C2-C3) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 16 ) hydrocarbyl, aliphatic (C2-C3) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 15 ) hydrocarbyl, aliphatic (C2-C3) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 14 ) hydrocarbyl, aliphatic (C2-C3) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 13 ) hydrocarbyl, aliphatic (C2-C3) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 12 ) hydrocarbyl, aliphatic (C2-C3) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 11 ) hydrocarbyl, aliphatic (C2-C3) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 10) hydrocarbyl, an aliphatic (C2-C9) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, an aliphatic (C2-C8) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, an aliphatic (C2-C7) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, an aliphatic (C2-C6) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, an aliphatic (C2-C5) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, an aliphatic (C2-C4) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, and an aliphatic (C2-C3) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons.

[0057] In various embodiments of the present invention, R is an aliphatic (C3-C6) optionally substituted with 1 or 2 phenyl groups, with the proviso that R contains 20 or fewer carbons. 20 ) hydrocarbyl, aliphatic (C3-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 19 ) hydrocarbyl, aliphatic (C3-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 18 ) hydrocarbyl, aliphatic (C3-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 17 ) hydrocarbyl, aliphatic (C3-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 16 ) hydrocarbyl, aliphatic (C3-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 15 ) hydrocarbyl, aliphatic (C3-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 14 ) hydrocarbyl, aliphatic (C3-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 13 ) hydrocarbyl, aliphatic (C3-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons12 ) hydrocarbyl, aliphatic (C3-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 11 ) hydrocarbyl, aliphatic (C3-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 10 ) hydrocarbyl, an aliphatic (C3-C9) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, an aliphatic (C3-C8) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, an aliphatic (C3-C7) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, an aliphatic (C3-C6) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, an aliphatic (C3-C5) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, and an aliphatic (C3-C4) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons.

[0058] In various embodiments of the present invention, R is an aliphatic (C4-C 20 ) hydrocarbyl, aliphatic (C4-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 19 ) hydrocarbyl, aliphatic (C4-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 18 ) hydrocarbyl, aliphatic (C4-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 17 ) hydrocarbyl, aliphatic (C4-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 16 ) hydrocarbyl, aliphatic (C4-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 15 ) hydrocarbyl, aliphatic (C4-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 14) hydrocarbyl, aliphatic (C4-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 13 ) hydrocarbyl, aliphatic (C4-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 12 ) hydrocarbyl, aliphatic (C4-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 11 ) hydrocarbyl, aliphatic (C4-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 10 ) hydrocarbyl, an aliphatic (C4-C9) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, with the proviso that R contains 20 or fewer carbons, an aliphatic (C4-C8) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, an aliphatic (C4-C7) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, an aliphatic (C4-C6) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, and an aliphatic (C4-C5) hydrocarbyl optionally substituted with 1 or 2 phenyl groups.

[0059] In various embodiments of the present invention, R is an aliphatic (C5-C 20 ) hydrocarbyl, aliphatic (C5-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 19 ) hydrocarbyl, aliphatic (C5-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 18 ) hydrocarbyl, aliphatic (C5-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 17 ) hydrocarbyl, aliphatic (C5-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 16 ) hydrocarbyl, aliphatic (C5-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 15) hydrocarbyl, aliphatic (C5-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 14 ) hydrocarbyl, aliphatic (C5-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 13 ) hydrocarbyl, aliphatic (C5-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 12 ) hydrocarbyl, aliphatic (C5-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 11 ) hydrocarbyl, aliphatic (C5-C6) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 10 ) hydrocarbyl, aliphatic (C5-C9) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, with the proviso that R contains 20 or fewer carbons, aliphatic (C5-C8) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, aliphatic (C5-C7) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, and aliphatic (C5-C6) hydrocarbyl optionally substituted with 1 or 2 phenyl groups.

[0060] In various embodiments of the present invention, R is an aliphatic (C6-C 20 ) hydrocarbyl, aliphatic (C6-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 19 ) hydrocarbyl, aliphatic (C6-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 18 ) hydrocarbyl, aliphatic (C6-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 17 ) hydrocarbyl, aliphatic (C6-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 16) hydrocarbyl, aliphatic (C6-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 15 ) hydrocarbyl, aliphatic (C6-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 14 ) hydrocarbyl, aliphatic (C6-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 13 ) hydrocarbyl, aliphatic (C6-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 12 ) hydrocarbyl, aliphatic (C6-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 11 ) hydrocarbyl, aliphatic (C6-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 10 ) hydrocarbyl, aliphatic (C6-C9) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, with the proviso that R contains 20 or fewer carbons, aliphatic (C6-C8) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, and aliphatic (C6-C7) hydrocarbyl optionally substituted with 1 or 2 phenyl groups.

[0061] In various embodiments of the present invention, R is an aliphatic (C7-C 20 ) hydrocarbyl, aliphatic (C7-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 19 ) hydrocarbyl, aliphatic (C7-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 18 ) hydrocarbyl, aliphatic (C7-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 17 ) hydrocarbyl, aliphatic (C7-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 16) hydrocarbyl, aliphatic (C7-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 15 ) hydrocarbyl, aliphatic (C7-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 14 ) hydrocarbyl, aliphatic (C7-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 13 ) hydrocarbyl, aliphatic (C7-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 12 ) hydrocarbyl, aliphatic (C7-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 11 ) hydrocarbyl, aliphatic (C7-C) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 10 ) hydrocarbyl, aliphatic (C7-C9) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, with the proviso that R contains 20 or fewer carbons, and aliphatic (C7-C8) hydrocarbyl optionally substituted with 1 or 2 phenyl groups.

[0062] In various embodiments of the present invention, R is an aliphatic (C8-C10) optionally substituted with 1 or 2 phenyl groups, with the proviso that R contains 20 or fewer carbons. 20 ) hydrocarbyl, aliphatic (C8-C10) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 19 ) hydrocarbyl, aliphatic (C8-C10) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 18 ) hydrocarbyl, aliphatic (C8-C10) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 17 ) hydrocarbyl, aliphatic (C8-C10) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 16) hydrocarbyl, aliphatic (C8-C10) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 15 ) hydrocarbyl, aliphatic (C8-C10) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 14 ) hydrocarbyl, aliphatic (C8-C10) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 13 ) hydrocarbyl, aliphatic (C8-C10) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 12 ) hydrocarbyl, aliphatic (C8-C10) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 11 ) hydrocarbyl, aliphatic (C8-C10) optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons 10 ) hydrocarbyl, and aliphatic (C8-C9) hydrocarbyl optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons.

[0063] In various embodiments of the present invention, R is an aliphatic (C9-C 20 ) hydrocarbyl, aliphatic (C9-C10) optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons 19 ) hydrocarbyl, aliphatic (C9-C10) optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons 18 ) hydrocarbyl, aliphatic (C9-C10) optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons 17 ) hydrocarbyl, aliphatic (C9-C10) optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons 16 ) hydrocarbyl, aliphatic (C9-C10) optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons 15 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C9-C 14) hydrocarbyl, optionally phenyl-substituted aliphatic (C9-C 13 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C9-C 12 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C9-C 11 ) hydrocarbyl, and optionally phenyl-substituted aliphatic (C9-C 10 ) hydrocarbyl.

[0064] In various embodiments of the present invention, R is an aliphatic (C 10 -C 20 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 10 -C 19 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 10 -C 18 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 10 -C 17 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 10 -C 16 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 10 -C 15 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C 10 -C 14 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C 10 -C 13 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C 10 -C 12 ) hydrocarbyl, and optionally phenyl-substituted aliphatic (C 10 -C 11 ) hydrocarbyl.

[0065] In various embodiments of the present invention, R is an aliphatic (C 11 -C 20 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 11 -C 19 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 11 -C 18 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 11 -C 17 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 11 -C 16 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 11 -C 15 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C 11 -C 14 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C 11 -C 13 ) hydrocarbyl, and optionally phenyl-substituted aliphatic (C 11 -C 12 ) hydrocarbyl.

[0066] In various embodiments of the present invention, R is an aliphatic (C 12 -C 20 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 12 -C 19 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 12 -C 18 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C12 -C 17 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 12 -C 16 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 12 -C 15 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C 12 -C 14 ) hydrocarbyl, and optionally phenyl-substituted aliphatic (C 12 -C 13 ) hydrocarbyl.

[0067] In various embodiments of the present invention, R is an aliphatic (C 13 -C 20 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 13 -C 19 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 13 -C 18 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 13 -C 17 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 13 -C 16 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 13 -C 15 ) hydrocarbyl, and optionally phenyl-substituted aliphatic (C 13 -C 14 ) hydrocarbyl.

[0068] In various embodiments of the present invention, R is an aliphatic (C14 -C 20 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 14 -C 19 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 14 -C 18 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 14 -C 17 ) hydrocarbyl, aliphatic optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 14 -C 16 ) hydrocarbyl, and optionally phenyl-substituted aliphatic (C 14 -C 15 ) hydrocarbyl.

[0069] In various embodiments of the present invention, R is an aliphatic (C 15 -C 20 ) Hydrocarbyl, Aliphatic (C 15 -C 19 ) Hydrocarbyl, Aliphatic (C 15 -C 18 ) Hydrocarbyl, Aliphatic (C 15 -C 17 ) hydrocarbyl, and aliphatic (C 15 -C 16 ) hydrocarbyl.

[0070] In various embodiments of the present invention, R is an aliphatic (C 16 -C 20 ) Hydrocarbyl, Aliphatic (C 16 -C 19 ) Hydrocarbyl, Aliphatic (C 16 -C 18 ) hydrocarbyl, and aliphatic (C 16 -C 17 ) hydrocarbyl.

[0071] In various embodiments of the present invention, R is an aliphatic (C 17 -C 20 ) Hydrocarbyl, Aliphatic (C 17 -C 19 ) hydrocarbyl, and aliphatic (C 17 -C 18 ) hydrocarbyl.

[0072] In various embodiments of the present invention, R is an aliphatic (C 18 -C 20 ) hydrocarbyl or aliphatic (C 18 -C 19 ) hydrocarbyl.

[0073] In various embodiments of the present invention, R is an aliphatic (C 19 -C 20 ) hydrocarbyl.

[0074] In various embodiments of the present invention, R is an aliphatic (C 20 ) Hydrocarbyl, Aliphatic (C 19 ) Hydrocarbyl, Aliphatic (C 18 ) Hydrocarbyl, Aliphatic (C 17 ) Hydrocarbyl, Aliphatic (C 16 ) Hydrocarbyl, Aliphatic (C 15 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C 14 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C 13 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C 12 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C 11 ) hydrocarbyl, optionally phenyl-substituted aliphatic (C 10) hydrocarbyl, an aliphatic (C9) hydrocarbyl optionally substituted with phenyl groups, an aliphatic (C8) hydrocarbyl optionally substituted with one or two phenyl groups, an aliphatic (C7) hydrocarbyl optionally substituted with one or two phenyl groups, an aliphatic (C6) hydrocarbyl optionally substituted with one or two phenyl groups, an aliphatic (C5) hydrocarbyl optionally substituted with one or two phenyl groups, an aliphatic (C4) hydrocarbyl optionally substituted with one or two phenyl groups, and an aliphatic (C3) hydrocarbyl optionally substituted with one or two phenyl groups.

[0075] In some embodiments of the invention, R is optionally substituted with 1 or 2 phenyl groups, with the proviso that R contains 20 or fewer carbons (C1-C 20 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C1-C 19 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C1-C 18 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C1-C 17 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C1-C 16 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C1-C 15 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C1-C 14 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C1-C 13 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C1-C 12 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C1-C 11) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C1-C 10 )alkyl, (C1-C9)alkyl optionally substituted with one or two phenyl groups, with the proviso that R contains 20 or fewer carbons, (C1-C8)alkyl optionally substituted with one or two phenyl groups, (C1-C7)alkyl optionally substituted with one or two phenyl groups, (C1-C6)alkyl optionally substituted with one or two phenyl groups, (C1-C5)alkyl optionally substituted with one or two phenyl groups, (C1-C4)alkyl optionally substituted with one or two phenyl groups, and (C1-C3)alkyl optionally substituted with one or two phenyl groups.

[0076] In various embodiments of the present invention, R is optionally substituted with one or two phenyl groups, with the proviso that R contains 20 or fewer carbons (C2-C 20 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C2-C 19 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C2-C 18 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C2-C 17 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C2-C 16 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C2-C 15 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C2-C 14 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C2-C 13 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C2-C 12) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C2-C 11 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C2-C 10 )alkyl, (C2-C9)alkyl optionally substituted with one or two phenyl groups, (C2-C8)alkyl optionally substituted with one or two phenyl groups, (C2-C7)alkyl optionally substituted with one or two phenyl groups, (C2-C6)alkyl optionally substituted with one or two phenyl groups, (C2-C5)alkyl optionally substituted with one or two phenyl groups, (C2-C4)alkyl optionally substituted with one or two phenyl groups, and (C2-C3)alkyl optionally substituted with one or two phenyl groups, provided that R contains 20 or fewer carbons.

[0077] In various embodiments of the present invention, R is optionally substituted with one or two phenyl groups, with the proviso that R contains 20 or fewer carbons (C3-C 20 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C3-C 19 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C3-C 18 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C3-C 17 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C3-C 16 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C3-C 15 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C3-C 14 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C3-C 13) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C3-C 12 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C3-C 11 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C3-C 10 )alkyl, (C3-C9)alkyl optionally substituted with one or two phenyl groups, (C3-C8)alkyl optionally substituted with one or two phenyl groups, (C3-C7)alkyl optionally substituted with one or two phenyl groups, (C3-C6)alkyl optionally substituted with one or two phenyl groups, (C3-C5)alkyl optionally substituted with one or two phenyl groups, (C3-C4)alkyl optionally substituted with one or two phenyl groups, provided that R contains 20 or fewer carbons.

[0078] In various embodiments of the present invention, R is optionally substituted with one or two phenyl groups, with the proviso that R contains 20 or fewer carbons (C4-C 20 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C4-C 19 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C4-C 18 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C4-C 17 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C4-C 16 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C4-C 15 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C4-C 14 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C4-C 13) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C4-C 12 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C4-C 11 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C4-C 10 )alkyl, (C4-C9)alkyl optionally substituted with one or two phenyl groups, with the proviso that R contains 20 or fewer carbons, (C4-C8)alkyl optionally substituted with one or two phenyl groups, (C4-C7)alkyl optionally substituted with one or two phenyl groups, (C4-C6)alkyl optionally substituted with one or two phenyl groups, and (C4-C5)alkyl optionally substituted with one or two phenyl groups.

[0079] In various embodiments of the present invention, R is optionally substituted with one or two phenyl groups, with the proviso that R contains 20 or fewer carbons (C5-C 20 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C5-C 19 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C5-C 18 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C5-C 17 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C5-C 16 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C5-C 15 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C5-C 14 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C5-C 13) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C5-C 12 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C5-C 11 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C5-C 10 )alkyl, (C5-C9)alkyl optionally substituted with one or two phenyl groups, with the proviso that R contains 20 or fewer carbons, (C5-C8)alkyl optionally substituted with one or two phenyl groups, (C5-C7)alkyl optionally substituted with one or two phenyl groups, and (C5-C6)alkyl optionally substituted with one or two phenyl groups.

[0080] In various embodiments of the present invention, R is optionally substituted with one or two phenyl groups, with the proviso that R contains 20 or fewer carbons (C6-C 20 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C6-C 19 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C6-C 18 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C6-C 17 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C6-C 16 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C6-C 15 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C6-C 14 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C6-C 13 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C6-C 12) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C6-C 11 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C6-C 10 )alkyl, (C6-C9)alkyl optionally substituted with 1 or 2 phenyl groups, with the proviso that R contains 20 or fewer carbons, (C6-C8)alkyl optionally substituted with 1 or 2 phenyl groups, and (C6-C7)alkyl optionally substituted with 1 or 2 phenyl groups.

[0081] In various embodiments of the present invention, R is optionally substituted with one or two phenyl groups, with the proviso that R contains 20 or fewer carbons (C7-C 20 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C7-C 19 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C7-C 18 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C7-C 17 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C7-C 16 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C7-C 15 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C7-C 14 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C7-C 13 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C7-C 12 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C7-C 11) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C7-C 10 )alkyl, (C7-C9)alkyl optionally substituted with 1 or 2 phenyl groups, with the proviso that R contains 20 or fewer carbons, and (C7-C8)alkyl optionally substituted with 1 or 2 phenyl groups.

[0082] In various embodiments of the present invention, R is optionally substituted with one or two phenyl groups, with the proviso that R contains 20 or fewer carbons (C8-C 20 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C8-C 19 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C8-C 18 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C8-C 17 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C8-C 16 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C8-C 15 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C8-C 14 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C8-C 13 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C8-C 12 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C8-C 11 ) alkyl, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C8-C 10)alkyl, and (C8-C9)alkyl optionally substituted with 1 or 2 phenyl groups, with the proviso that R contains 20 or fewer carbons.

[0083] In various embodiments of the present invention, R is an optionally substituted phenyl group, with the proviso that R contains 20 or fewer carbons (C9-C 20 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C9-C 19 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C9-C 18 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C9-C 17 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C9-C 16 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C9-C 15 ) alkyl, optionally substituted with a phenyl group (C9-C 14 ) alkyl, optionally substituted with a phenyl group (C9-C 13 ) alkyl, optionally substituted with a phenyl group (C9-C 12 ) alkyl, optionally substituted with a phenyl group (C9-C 11 ) alkyl, and optionally phenyl substituted (C9-C 10 ) alkyl.

[0084] In various embodiments of the present invention, R is an optionally substituted phenyl group, with the proviso that R contains 20 or fewer carbons (C 10 -C 20 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 10 -C 19 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 10 -C 18 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C10 -C 17 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 10 -C 16 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 10 -C 15 ) alkyl, optionally substituted with a phenyl group (C 10 -C 14 ) alkyl, optionally substituted with a phenyl group (C 10 -C 13 ) alkyl, optionally substituted with a phenyl group (C 10 -C 12 ) alkyl, and optionally phenyl substituted (C 10 -C 11 ) alkyl.

[0085] In various embodiments of the present invention, R is an optionally substituted phenyl group, with the proviso that R contains 20 or fewer carbons (C 11 -C 20 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 11 -C 19 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 11 -C 18 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 11 -C 17 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 11 -C 16 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 11 -C 15 ) alkyl, optionally substituted with a phenyl group (C 11 -C 14 ) alkyl, optionally substituted with a phenyl group (C 11 -C 13 ) alkyl, and optionally phenyl substituted (C11 -C 12 ) alkyl.

[0086] In various embodiments of the present invention, R is an optionally substituted phenyl group, with the proviso that R contains 20 or fewer carbons (C 12 -C 20 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 12 -C 19 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 12 -C 18 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 12 -C 17 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 12 -C 16 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 12 -C 15 ) alkyl, optionally substituted with a phenyl group (C 12 -C 14 ) alkyl, and optionally phenyl substituted (C 12 -C 13 ) alkyl.

[0087] In various embodiments of the present invention, R is an optionally substituted phenyl group, with the proviso that R contains 20 or fewer carbons (C 13 -C 20 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 13 -C 19 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 13 -C 18 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 13 -C 17) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 13 -C 16 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 13 -C 15 ) alkyl, and optionally phenyl substituted (C 13 -C 14 ) alkyl.

[0088] In various embodiments of the present invention, R is an optionally substituted phenyl group, with the proviso that R contains 20 or fewer carbons (C 14 -C 20 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 14 -C 19 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 14 -C 18 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 14 -C 17 ) alkyl, optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 14 -C 16 ) alkyl, and optionally substituted with phenyl groups, provided that R contains 20 or fewer carbons (C 14 -C 15 ) alkyl.

[0089] In various embodiments of the present invention, R is 15 -C 20 ) alkyl, (C 15 -C 19 ) alkyl, (C 15 -C 18 ) alkyl, (C 15 -C 17 ) alkyl, and (C 15 -C 16 ) alkyl.

[0090] In various embodiments of the present invention, R is 16 -C 20 ) alkyl, (C 16 -C 19 ) alkyl, (C 16 -C 18 ) alkyl, and (C 16 -C 17 ) alkyl.

[0091] In various embodiments of the present invention, R is 17 -C 20 ) alkyl, (C 17 -C 19 ) alkyl, and (C 17 -C 18 ) alkyl.

[0092] In various embodiments of the present invention, R is (C 18 -C 20 ) alkyl or (C 18 -C 19 ) alkyl.

[0093] In various embodiments of the present invention, R is (C 19 -C 20 ) alkyl.

[0094] In various embodiments of the present invention, R is 20 ) alkyl, (C 19 ) alkyl, (C 18 ) alkyl, (C 17 ) alkyl, (C 16 ) alkyl, (C 15 ) alkyl, optionally substituted with a phenyl group (C 14 ) alkyl, optionally substituted with a phenyl group (C 13 ) alkyl, optionally substituted with a phenyl group (C 12 ) alkyl, optionally substituted with a phenyl group (C 11 ) alkyl, optionally substituted with a phenyl group (C 10 ) alkyl, (C9)alkyl optionally substituted with phenyl groups, (C8)alkyl optionally substituted with one or two phenyl groups, (C7)alkyl optionally substituted with one or two phenyl groups, (C6)alkyl optionally substituted with one or two phenyl groups, (C5)alkyl optionally substituted with one or two phenyl groups, (C4)alkyl optionally substituted with one or two phenyl groups, (C3)alkyl optionally substituted with one or two phenyl groups, ethyl optionally substituted with one or two phenyl groups, and methyl optionally substituted with one or two phenyl groups.

[0095] In various embodiments of the present invention, R is C n H mIn some of the above embodiments, n is 1 and m is 3 (i.e., methyl). In some of the above embodiments, n is 2 and m is 5 (i.e., ethyl). In some of the above embodiments, n is 3 and m is selected from 3, 5, and 7. In some of the above embodiments, n is 4 and m is selected from 5, 7, and 9. In some of the above embodiments, n is 5 and m is selected from 7, 9, and 11. In some of the above embodiments, n is 6 and m is selected from 5, 7, 9, 11, and 13. In some of the above embodiments, n is 7 and m is selected from 7, 9, 11, 13, and 15. In some of the above embodiments, n is 8 and m is selected from 5, 7, 9, 11, 13, 15, and 17. In some of the above embodiments, n is 9 and m is selected from 7, 9, 11, 13, 15, 17, and 19. In some of the above embodiments, n is 10 and m is selected from 7, 9, 11, 13, 15, 17, 19, and 21. In some of the above embodiments, n is 11 and m is selected from 9, 11, 13, 15, 17, 19, 21, and 23. In some of the above embodiments, n is 12 and m is selected from 7, 9, 11, 13, 15, 17, 19, 21, 23, and 25. In some of the above embodiments, n is 13 and m is selected from 9, 11, 13, 15, 17, 19, 21, 23, 25, and 27. In some of the above embodiments, n is 14 and m is selected from 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, and 29. In some of the above embodiments, n is 15 and m is selected from 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, and 31. In some of the above embodiments, n is 16 and m is selected from 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, and 33. In some of the above embodiments, n is 17 and m is selected from 11, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, and 35.In some of the above embodiments, n is 18 and m is selected from 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, and 37. In some of the above embodiments, n is 19 and m is selected from 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, and 39. In some of the above embodiments, n is 20 and m is selected from 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, and 41.

[0096] In some embodiments, R is n-propyl, isopropyl, cyclopropyl, n-butyl, 1-methylpropyl, 2-ethylbutyl, 1-methyl-2-ethylbutyl, 2-methylpropyl, tert-butyl, 2-methylcyclopropyl, 1-methylcyclopropyl, cyclobutyl, cyclopropylmethyl (i.e., [ka] ), n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, cyclobutylmethyl (i.e., [ka] ), 2-(cyclopropyl)ethyl (i.e. [ka] ), cyclopentyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1,1,2-trimethylpropyl, 1,2,2-trimethylpropyl, 3-(cyclopropyl)propyl (i.e., [ka] ), 2-(cyclobutyl)ethyl (i.e. [ka] ), cyclopentylmethyl (i.e. [ka] ), cyclohexyl, cyclohexylmethyl, 2-cyclohexylethyl, dicyclohexylmethyl, n-octyl, benzyl, diphenylmethyl, decyl, dodecyl, tetradecyl, hexadecyl, hexadec-9-enyl, octadecyl, octadec-9-enyl, octadeca-9,12-dienyl, 2-propylpentyl, 2-butylhexyl, 2-pentylheptyl, 2-hexyloctyl.

[0097] As used herein, and as would be understood by one of skill in the art, reference to a "compound" is intended to include salts of that compound, unless expressly limited further.

[0098] The term "pharmaceutically acceptable salts" refers to salts whose counterions are derived from pharmaceutically acceptable non-toxic acids and bases. Suitable pharmaceutically acceptable acids for the salts of the amino-substituted compounds of the invention include, for example, acetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid (besylic acid), benzoic acid, boric acid, lactic acid, camphoric acid, camphorsulfonic acid, carbonic acid, citric acid, ethanedisulfonic acid, ethanesulfonic acid, ethylenediaminetetraacetic acid, formic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, iodine acid, and the like. Hydrogen, hydroxynaphthoic, isethionic, lactic, lactobionic, laurylsulfonic, maleic, malic, mandelic, methanesulfonic, mucic, naphthylenesulfonic, nitric, oleic, pamoic, pantothenic, phosphoric, pivalic, polygalacturonic, salicylic, stearic, succinic, sulfuric, tannic, tartaric, teoclatic, p-toluenesulfonic, etc. Suitable pharma- ceutically acceptable base addition salts for the carboxylic acid-substituted compounds of the invention include, but are not limited to, metallic salts made from aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc, or organic salts made from lysine, arginine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine. Further pharma-ceutically acceptable salts include, where appropriate, non-toxic ammonium cations, and carboxylate, sulfonate, and phosphonate anions attached to alkyl groups having 1 to 20 carbon atoms.

[0099] It will be appreciated that the compounds of the present invention may exist in isotopically labeled form, i.e., the compounds may contain one or more atoms with an atomic mass or mass number different from the atomic mass or mass number normally found in nature. As another example, multiple molecules of a single structure may contain at least one atom that exists in an isotopic abundance different from the isotopic abundance found in nature. Radioactive isotopes of hydrogen, carbon, phosphorus, fluorine, chlorine and iodine include, respectively: 2H, 3 H, 11 C. 13 C. 14 C. 15 N, 35 S, 18 F, 36 Cl, 125 I, 124 I and 131 I. Compounds containing these radioisotopes and / or other radioisotopes of other atoms are within the scope of the present invention. Deuterium has been used to improve the metabolic stability of compounds, and those skilled in the art will recognize that the principles can be applied to these compounds. The radioisotope tritium, i.e. 3 H and carbon 14 i.e. 14 C is particularly preferred because it is easy to prepare and detectable. 11 C. 13 N, 15 O. 124 I, and 18 Compounds containing F are well suited for positron emission tomography. The radioisotope-labeled compounds of formula I and II of the present invention and their prodrugs can generally be prepared by methods well known to those skilled in the art. Conveniently, such radioisotope-labeled compounds can be prepared by carrying out the procedures disclosed in the examples and schemes, by using readily available radioisotope-labeled reagents instead of non-radioisotope-labeled reagents.

[0100] Although the present invention can be embodied in various forms, the preferred embodiment of the present invention is shown. It should be understood, however, that the present disclosure should be regarded as an exemplification of the principles of the present invention, and is not intended to limit the present invention to the illustrated embodiment. During examination, certain members of a genus described in the claims may be found to be unlicensed to the inventors in this application. In this case, the subsequent exclusion of certain species from the scope of the applicant's claims should be considered an artifact of patent examination and not a reflection of the inventors' conception or description of the invention; the invention encompasses all members of genus I that have not yet been made public.

[0101] The terms "subject" or "subject in need thereof" are used interchangeably herein. These terms refer to a patient who has been diagnosed with an underlying disease to be treated. The subject may be currently experiencing or may have previously experienced symptoms associated with the disease. Additionally, a "subject in need thereof" may be a patient who is at risk of developing a particular disease, or who suffers from one or more of the physiological systems of a disease, even if the disease has not been diagnosed. As a non-limiting example, a "subject in need thereof," for purposes of this application, may include a subject who is currently diagnosed with or has been previously diagnosed with epilepsy, or who is at risk for epilepsy seizures regardless of current symptomatology.

[0102] As used herein, the terms "treatment" and "treating" are used interchangeably. These terms refer to an approach to obtain a beneficial or desired result, such as, but not limited to, a therapeutic benefit. A therapeutic benefit includes eradicating or alleviating the underlying disease being treated; and further includes eradicating or alleviating one or more symptoms associated with the underlying disease, such that the patient observes an improvement even if the patient is still afflicted by the underlying disease.

[0103] The compounds described herein are useful for treating epilepsy and pain. When referring to "treating epilepsy," the term includes the relief of symptoms of seizures and convulsions. When referring to "treating pain," the term includes the relief of neuropathic pain, chemotherapy-induced pain, back pain, bone pain, abdominal pain, post-operative pain, traumatic pain, menstrual pain, muscle pain, joint pain, headache, migraine, dental pain, induced pain, and inflammatory pain.

[0104] [Chemical synthesis of compounds] In general, the compounds of the present invention can be prepared by the methods illustrated in the general reaction schemes, for example as described below, or by modifications thereof, using readily available starting materials, reagents and conventional synthetic procedures. In these reactions, it is also possible to utilize variants that are known per se but not mentioned herein. The starting materials may be commercially available, synthesized as described in the examples, or obtained by methods well known to those skilled in the art. Standard synthetic methods and procedures for the preparation of organic molecules and functional group transformations and manipulations can be readily obtained from the relevant scientific literature or from standard textbooks in the field. It is understood that where typical or preferred reaction process conditions (i.e., reaction temperatures, times, molar ratios of reactants, solvents, pressures, etc.) are given; other reaction process conditions can also be used unless otherwise specified. Optimum reaction conditions may vary depending on the particular reactants or solvents used. Those skilled in the art will recognize that the nature and order of the synthetic steps presented can be varied for the purpose of optimizing the formation of the compounds described. A comprehensive list of abbreviations used by organic chemists appears in the first issue of each volume of the Journal of Organic Chemistry. The above list, typically presented in a table entitled "Standard List of Abbreviations", provides definitions of abbreviations not included in the abbreviation list below. ℃ Celsius temperature ACN Acetonitrile Cl Chloride Cs2CO3 Cesium carbonate DCC N,N'-dicyclohexylcarbodiimide DCM dichloromethane or methylene chloride DIPEA Diisopropylethylamine DMAP 4-Dimethylaminopyridine or N,N-Dimethylaminopyridine DMF N,N-Dimethylformamide EDC N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride eq. equivalent weight EtOAc Ethyl acetate EtOH Ethanol ESI Electrospray Ionization g grams HATU 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate HBTU 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate HCl Hydrochloric acid HPLC High Performance Liquid Chromatography h time I iodide LC / MS Liquid Chromatography Mass Spectrometry LDA Lithium diisopropylamide LiHMDS Lithium bis(trimethylsilyl)amide K2CO3 Potassium Carbonate KHMDS Potassium bis(trimethylsilyl)amide KMnO4 Potassium Permanganate M Molar concentration μm microns MeOH Methanol mg milligram min mL Milliliters mmol N specified concentration Na2CO3 Sodium Carbonate NaH Sodium hydride NaHCO3 Sodium bicarbonate NaHMDS Sodium bis(trimethylsilyl)amide NaOH Sodium hydroxide NMR nuclear magnetic resonance OMs mesylate OTf Triflate OTs Tosylate Pd / C Palladium Carbon Ph Phenyl psi pounds per square inch PyBOP (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate Ru / C Ruthenium Carbon TBTU 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylaminium tetrafluoroborate TEA Triethylamine TFA Trifluoroacetic acid THF Tetrahydrofuran TMS-I Iodotrimethylsilane TLC Thin Layer Chromatography

[0105] The terms "protect", "deprotect" and "protected" for functionalities are used throughout this application. Such terms are well understood by those skilled in the art and are used in the context of a reaction process involving sequential treatment with a series of reagents. In that context, a protecting group refers to a group used to mask a functional group in a reaction process step where it is not desired that the reaction occur, but the functional group would otherwise react. The protecting group prevents reaction in that step, but can be removed thereafter to reveal the original functional group. Removal or "deprotection" occurs after the reaction or reactions that the functional group interferes with are complete. Thus, when a series of reagents is specified as in the reaction processes described herein, one skilled in the art can readily envision groups that would be suitable as "protecting groups". Groups suitable for that purpose are discussed in standard textbooks in chemistry such as TW Greene and PGM Wuts, "Protective Groups in Organic Synthesis", John Wiley and Sons, New York, 1999.

[0106] List of protecting groups and their abbreviations: Acetyl (Ac) Acylal Carboaryloxy (Alloc) Benzoyl (Bz) Benzyl (Bn, Bnl) Benzyl ester Carbamate Carbobenzyloxy (Cbz) Dimethoxytrityl, [bis-(4-methoxyphenyl)phenylmethyl] (DMT) Zitian Ethoxyethyl ether (EE) Fluorenylmethyloxycarbonyl (Fmoc) Methoxymethyl ether (MOM) Methoxytrityl [(4-methoxyphenyl)diphenylmethyl], MMT) Methyl ether Methyl (Me) Methyl ester Methylthiomethyl ether Orthoesters Oxazoline Pivaloyl (Piv) Phthalimide p-Methoxybenzylcarbonyl (Moz or MeOZ) p-Methoxybenzyl (PMB) p-Methoxyphenyl (PMP) Propargyl Alcohol Silyl groups (e.g. trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS), tri-iso-propylsilyloxymethyl (TOM) and triisopropylsilyl (TIPS)) Silyl Ester tert-Butyl ester tert-Butyloxycarbonyl (Boc or tBoc) Tetrahydropyranyl (THP) Tosyl (Ts or Tos) Trichloroethyl chloroformate (Troc) Trimethylsilylethoxymethyl (SEM) Trityl (triphenylmethyl, Tr) β-Methoxyethoxymethyl ether (MEM) (4-Nitrophenyl)sulfonyl or (4-Nitrophenyl)(dioxide)-lambda(6)-sulfanyl (Nosyl) 2-Cyanoethyl 2-Nitrophenylsulfenyl (Nps) 3,4-Dimethoxybenzyl (DMPM) 2,2,4,6,7-Pentamethyldihydrobenzofuran-5-sulfonyl (Pbf)

[0107] I. General Chemical Synthesis Section The compounds of the present invention are prepared using the methods illustrated in the general synthetic schemes and the experimental procedures detailed below. These general synthetic schemes and experimental procedures are presented for illustrative purposes and are not intended to be limiting. The starting materials used to prepare the compounds of the present invention are either commercially available or can be prepared using routine methods known in the art. Where indicated, compound names were generated using InstantJChem v6.1 for ChemAxon desktop and the IUPAC Naming plug-in.

[0108] The intermediate compounds I, V, and VIII have been previously described in the literature (U.S. Pat. No. 5,284,957 (B. Huff “Excitatory amino acid receptor antagonists,” 1994); U.S. Pat. No. 5,648,492 (AM Brian Arnold et al., “Process for preparing isoquinoline compounds,” 1997); Paul L. Ornstein et al., “(3SR,4aRS,6RS,8aRS)-6-[2-(1H-tetrazol-5-yl)ethyl)] decahydroisoquinoline-3-carboxylic Acid: A Structurally Novel, Systemically Active, Competitive AMPA Receptor Antagonist,” J. Med. Chem., (USA), 1993, Vol. 36, p. 2046-2048; Paul L. Ornstein et al., “Syntheses of Oxodecahydroisoquinoline-3-carboxylates. Useful Intermediates for The compound is prepared using the protocol described in “The Preparation of Conformationally Defined Excitatory Amino Acid Antagonists,” J. Org. Chem., (USA), 1991, Vol. 56, p. 4388-4392).

[0109] Scheme 1 illustrates a general synthetic scheme for preparing the desired compound IV from ketone I. In step 1, the carbamate-protected carboxylic acid I is treated with iodotrimethylsilane at room temperature, or alternatively with 6N hydrochloric acid at 90° C. to remove the carbamate-methyl protecting group and provide the ketone amino acid II. Compound II is reacted with benzyl chloroformate (Cbz), di-tert-butyl dicarbonate (Boc), or a similar reagent under basic conditions (e.g., 2N aqueous NaOH, triethylamine, diisopropylethylamine, etc.) to provide the carbamate-protected carboxylic acid III. Compound III is esterified to produce the ketone carbamate ester IV. Several methods are possible, including: (1) compound III is esterified with an alcohol (HO-R1) using a coupling reagent (e.g., N,N'-dicyclohexylcarbodiimide (DCC), EDC, HBTU, HATU, PyBOP, etc.) under basic conditions (e.g., triethylamine, diisopropylethylamine, pyridine, N,N-4-dimethylaminopyridine, etc.); (2) compound III is alkylated with an activated alkane (X-R1, where X = OTf, OTs, OMs, I, Br, and Cl) under basic conditions (e.g., NaOH, NaH, NaCO3, K2CO3, Cs2CO3, NaHCO3, etc.); and (3) compound III is esterified with an alcohol (HO-R1) using Mitsunobu conditions (e.g., diethylazodicarboxylate and triphenylphosphine, or similar reagents).

[0110] Scheme 1. General synthesis of ketone carbamate ester IV from ketone I. [ka]

[0111] Scheme 2 illustrates a general synthetic scheme for preparing the desired compound III from a ketone ester V. In step 1, the methyl carbamate-protected ethyl ester V is treated with iodotrimethylsilane at room temperature to remove the methyl carbamate protecting group and give the amino ester VI. The ethyl ester VI is reacted under basic conditions (e.g., 2N aqueous NaOH, triethylamine, diisopropylethylamine, etc.) with benzyl chloroformate (Cbz), di-tert-butyl dicarbonate (Boc), or a similar protecting group that is stable under basic conditions but more easily removable than methyl carbamate, to give the carbamate-protected ethyl ester VII. Compound VII is then hydrolyzed under standard aqueous basic conditions (e.g., 2N aqueous NaOH in an alcoholic solvent or similar aqueous-miscible organic solvent) to give the carbamate-protected acid III. Compound III is then esterified as described in Scheme 1 to produce the ketone carbamate ester IV.

[0112] Scheme 2. General synthesis of ketone carbamate acids III from ketone esters V. [ka]

[0113] Scheme 3 illustrates a general synthetic scheme for preparing the desired compound IX from the protected ketone carbamate ester IV. The tetrazole Wittig reagent VIII (previously prepared as described in B. Huff, U.S. Pat. No. 5,284,957) was deprotonated under strong basic conditions (e.g., LiHMDS, NaHMDS, LDA, etc.) and reacted with the ketone IV to prepare the olefin compound IX.

[0114] Scheme 3. General synthesis of olefin compounds IX from ketone carbamate esters IV. [ka] Scheme 4 illustrates a general synthetic scheme for preparing the desired compound X from a protected olefin IX. Free radical hydrofluorination of IX gives a mixture of fluorinated carbamate esters X (Timothy J. Barker and Dale L. Boger, "Fe(III) / NaBH4-Mediated Free Radical Hydrofluorination of Unactivated Alkenes", J. Am. Chem. Soc., (USA), 2012, Vol. 134, No. 33, p.13588-13591). The isomers can be separated by crystallization or chromatography to give isomerically pure fluorinated carbamate esters X.In addition, other exemplary hydrofluorination conditions, such as but not limited to a protocol using KHSO4-13HF complex (Zhichao Lu, Xiaojun Zeng, Gerald B. Hammond, and Bo Xu, “Widely Applicable Hydrofluorination of Alkenes via Bifunctional Activation of Hydrogen Fluoride,” J. Am. Chem. Soc., (USA), 2017, Vol. 139, p.18202-18205), a protocol using cobalt fluoride complex (Hiroki Shigehisa, Eriko Nishi, Mayu Fujisawa, and Kou Hiroya, “Cobalt-Catalyzed Hydrofluorination of Unactivated Olefins: A Radical Approach of Fluorine Transfer.” Org. Lett., (USA), 2013, Vol. 15, No. 20, 5158), a protocol using superacid HF / SbF5 (Sebastien Thibaudeau, Agnes Martin-Mingot, Marie-Paule Jouannetaud, Omar Karamb, and Fabien Zuninob, “A novel, facile route to beta-fluoroamines by hydrofluorination using superacid HF / SbF5”, Chem. Commun., (Eng), 2007, Vol. 31, p.98), and a protocol using HF-pyridine (George A. Olah and Michael Watkins, “Fluorinations With Pyridinium Polyhydrogen Fluoride”, Org. Synth., 1978, Vol. 58, p.75).

[0115] Scheme 4. General synthesis of fluorinated carbamate ester X from olefin IX [ka]

[0116] Scheme 5 illustrates a general synthetic scheme for making fluorinated ester amine prodrugs IX from fluorinated carbamate esters X. The carbamate protecting group is removed using established published protocols (e.g., hydrogenation using hydrogen gas and Pd / C catalyst for Cbz, TFA conditions or 4N HCl for Boc, etc.). Isomers can be separated by crystallization or chromatography to give isomerically pure fluorinated ester amines XI.

[0117] Scheme 5. General synthesis of fluorinated ester amine prodrugs XI from fluorinated carbamate esters X. [ka] Scheme 6 illustrates a general synthetic scheme for making the desired fluorinated amino acid compound 1 from the fluorinated amino ester XI. The ester is hydrolyzed under standard aqueous basic conditions (e.g., 2N aqueous NaOH or 2N aqueous NaOH in an alcoholic solvent or similar aqueous-miscible organic solvent) to give the fluorinated amino acid XII.

[0118] Scheme 6. General synthesis of fluorinated decahydroisoquinoline amino acids XII [ka] Final purification and isolation affords the fluorinated decahydroisoquinoline amino acids 1 and 2. [ka]

[0119] II. Experimental Chemical Synthesis Section All chemical reagents were purchased commercially and used without further purification. Reactions were carried out under air / nitrogen atmosphere according to the required conditions. Column chromatography was performed on silica gel 60 (230-400 mesh) and analytical TLC was carried out on silica gel coated plates. TLC plates were stained with cerium ammonium molybdate (CAM), p-anisaldehyde (Anis), potassium permanganate (KMnO4), or ninhydrin stain. 11 H NMR spectra were recorded using a Bruker 300 MHz or a Varian 300 MHz instrument using deuterium oxide, chloroform-d or methanol-d4 as solvent. HPLC spectra were recorded using an Agilent 1100 series HPLC with a Zorbax SB-C18 (4.6 x 150 mm) column with a gradient elution of 5% B → 95% B in 8.5 min (mobile phase A: 0.05% HClO4 in water; mobile phase B: acetonitrile) and UV detection at 205 nm, or a Waters Sunfire C18 (4.6 x 75 mm, 3.5 μm, part number 186002552) column with a gradient elution of 5% B → 95% B in 8.6 min (mobile phase A: 0.1% TFA in water; mobile phase B: 0.1% TFA in acetonitrile) and UV detection at all wavelengths. Mass spectrometry was performed using an Advion ExpressionCMS® (ESI) or Agilent (Hewlett Packard 1100 series MSD) with a MassLynx™ interface (ESI: positive or negative ion mode) or a Waters 29996, Micromass® ZQ (ESI: positive or negative ion mode). Preparative reversed phase chromatography was performed using a Gilson system equipped with a Waters Sunfire C18 OBD preparative column (30×150 mm column, 10 μm, part number 186002670). In some cases, normal phase silica gel column chromatography was performed using a Teledyne ISCO CombiFlash® system.

[0120] Preparation of ethyl (3S,4aS,8aR)-6-oxo-decahydroisoquinoline-3-carboxylate (I-01) [ka] To a solution of 3-ethyl 2-methyl (3S,4aS,8aR)-6-oxo-decahydroisoquinoline-2,3-dicarboxylate (35.42 g, 125 mmol) in methylene chloride (600 mL) (under nitrogen atmosphere) was added iodotrimethylsilane (100 g, 500 mmol) in one portion at room temperature. The reaction mixture was stirred overnight and quenched with ethanol (250 mL). The solution was concentrated under vacuum and dried under reduced pressure for 3 hours to give the desired crude aminoester I-01 as a yellow-orange solid (43 g crude), which was used directly in the next step without purification. 1 H NMR (300.13 MHz, CD3OD) δ 4.31 (q, J = 5.3 Hz, 2H), 4.17 (d, J = 9.6 Hz, 1H), 3.31-3.21 (m, 1H), 3.14 (dd, J = 9.6, 3.2 Hz, 1H), 2.21 (d, J = 9.5 Hz, 1H), 2.13-2.08 (m, 2H), 2.00 (dt, J = 10.1, 3.1 Hz, 1H), 1.88-1.69 (m, 4H), 1.58 (d, J = 9.9 Hz, 1H), 1.40-1.35 (m, 1H), 1.32 (t, J = 5.3 Hz, 3H) ppm. Scale-up batch: To a solution of 3-ethyl 2-methyl (3S,4aS,8aR)-6-oxo-decahydroisoquinoline-2,3-dicarboxylate (74.4 g, 262.6 mmol) in methylene chloride (1200 mL) (under nitrogen atmosphere) was added iodotrimethylsilane (200 g, 1.0 mol) in one portion at room temperature. The reaction mixture was stirred overnight and quenched with ethanol (280 mL). The solution was concentrated under vacuum and dried under reduced pressure for 3 hours to give the desired crude aminoester I-01 as a yellow-orange solid (90.5 g crude), which was used directly without further purification.

[0121] <Synthesis of ketone carbamate ester IV> Preparation of 2-benzyl 3-ethyl (3S,4aS,8aR)-6-oxo-decahydroisoquinoline-2,3-dicarboxylate (I-02) [ka] To a slurry of I-01 (crude, 6.25 mmol) in methylene chloride (50 mL) was added triethylamine (3.5 mL, 25.1 mmol) at 5-10 °C and the mixture was stirred under nitrogen atmosphere for 10 min. Benzyl chloroformate (1.12 mL, 7.62 mmol) was added slowly at 5-10 °C. The mixture was warmed to room temperature and stirred for 2-3 h (the reaction was monitored by TLC and KMnO4 staining). The mixture was adjusted to pH 3-4 with 3N HCl and diluted with ethyl acetate (50 mL). The layers were separated and the combined organic layers were washed with brine (15 mL), dried over sodium sulfate, and concentrated under vacuum. The resulting residue was purified by flash column chromatography (silica gel, 0% to 40% ethyl acetate / hexanes) to give I-02 as a pale yellow oil (2.12 g, 94% yield). 1 H NMR (300.13 MHz, CDCl3) δ 7.36-7.27 (m, 5H), 5.14 (d, J = 24.6, 14.1 Hz, 2H), 4.92 (dd, J = 41.3, 8.3 Hz, 1H), 4.22-4.14 (m, 2H), 4.02 (dd, J = 22.5, 13.5 Hz, 1H), 3.27 (ddd, J = 34.5, 13.5, 3.0 Hz, 1H), 2.59 (dd, J = 16.5, 6.0 Hz, 1H), 2.38-1.65 (m, 9H), 1.26 (t, J = 7.2 Hz, 1.5H), 1.21 (t, J = 7.2 Hz, 1.5H) ppm.

[0122] Preparation of 2-tert-butyl 3-ethyl (3S,4aS,8aR)-6-oxo-decahydroisoquinoline-2,3-dicarboxylate (I-03) [ka] Crude I-01 (125 mmol) from the previous reaction was dissolved in methylene chloride (600 mL) and triethylamine (60.7 mL, 435 mmol) was added. After stirring for 15 min, a solution of di-tert-butyl dicarbonate (32.7 g, 150 mmol) in methylene chloride (100 mL) was added. The resulting mixture was stirred overnight at room temperature and then concentrated under vacuum. The resulting solid was suspended in ethyl acetate (300 mL) and filtered. The filtrate was washed with 1N HCl (60 mL) and brine (100 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated under vacuum. The resulting residue was purified by flash column chromatography (silica gel, 0%→25% ethyl acetate / hexanes) to give I-03 as a colorless oil (30.0 g, 74% yield). 1 H NMR (300.13 MHz, CDCl3) δ 4.82 (dd, J = 65.0, 3.9 Hz, 1H), 4.17-4.14 (m, 2H), 3.90 (dd, J = 27.9, 10.2 Hz, 1H), 3.15 (dd, J = 39.7, 10.1 Hz, 1H), 2.56 (dd, J = 10.7, 4.3 Hz, 1H), 2.41-2.30 (m, 2H), 2.16-1.97 (m, 5H), 1.95-1.78 (m, 1H), 1.75-1.65 (m, 1H), 1.42 (s, 4.5H), 1.40 (s, 4.5H), 1.23 (t, J = 4.3 Hz, 3H) ppm. Scale-up batch: Crude I-01 (393.9 mmol) from the previous reaction was dissolved in methylene chloride (1.8 L) and triethylamine (219.3 mL, 1573 mmol) was added. After stirring for 15 min, a solution of di-tert-butyl dicarbonate (343.3 g, 1573 mmol) in methylene chloride (300 mL) was added. The resulting mixture was stirred overnight at room temperature and then concentrated under vacuum. The resulting solid was suspended in ethyl acetate (900 mL) and filtered. The filtrate was washed with 1N HCl (180 mL) and brine (100 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated under vacuum. The resulting colorless oil (151 g crude, quantitative yield) was used directly without further purification.

[0123] Preparation of (3S,4aS,8aR)-2-[(tert-butoxy)carbonyl]-6-oxo-decahydroisoquinoline-3-carboxylic acid (I-04) [ka] To a solution of I-03 (35.2 g, 108 mmol) in THF (100 mL) was added 2N NaOH (486 mL, 972 mmol) at room temperature under nitrogen atmosphere. The solution was stirred at room temperature for 24 h and then concentrated under vacuum to remove most of the THF. The aqueous layer was extracted with MTBE (3×150 mL) to remove organic impurities, acidified to pH ∼2 with 1N HCl, and extracted with ethyl acetate (4×300 mL). The combined organic layers were washed with brine (250 mL), dried over sodium sulfate, and concentrated under vacuum to give I-04 as a white foamy solid (24.9 g, 78% yield). 1 H NMR (300.13 MHz, CDCl3) δ 4.92 (d, J = 58.3 Hz, 1H), 3.95 (dd, J = 29.7, 10.1 Hz, 1H), 3.21 (dd, J = 32.9, 10.2 Hz, 1H), 2.61 (d, J = 10.2 Hz, 1H), 2.42-1.71 (m, 9H), 1.46 (s, 4.5H), 1.44 (s, 4.5H) ppm. Scale-up batch: To a solution of I-03 (151 g crude, 393.9 mmol) in THF (360 mL) was added 2N NaOH (1772 mL, 3545 mmol) at room temperature under nitrogen atmosphere. The solution was stirred at room temperature for 24 h and then concentrated under vacuum to remove most of the THF. The aqueous layer was extracted with MTBE (3×250 mL) to remove organic impurities, acidified to pH ∼2 with 1N HCl, and extracted with ethyl acetate (4×600 mL). The combined organic layers were washed with brine (500 mL), dried over sodium sulfate, and concentrated under vacuum to give I-04 as a white foamy solid (122 g crude, quantitative yield).

[0124] Preparation of (3S,4aS,8aR)-2-[(tert-butoxy)carbonyl]-6-oxo-decahydroisoquinoline-3-carboxylic acid (I-05) [ka] To a solution of I-04 (24.9 g, 83.7 mmol) in DMF (110 mL), solid NaHCO3 (49.4 g, 588 mmol) and 3-(iodomethyl)pentane (23.33 g, 110 mmol) were added at room temperature under nitrogen atmosphere and the mixture was stirred at 35-40 °C for 4 h. The reaction was monitored by HPLC and after completion of the reaction, the mixture was filtered and the solid was washed with acetonitrile (400 mL). The combined organic layers were concentrated and the crude residue obtained was redissolved in ethyl acetate (500 mL). The solution was washed with water (300 mL), brine (300 mL), dried over sodium sulfate and concentrated under vacuum. The residue was purified by flash column chromatography (silica gel, 0%→40% ethyl acetate / hexanes) to give I-05 as a red semi-solid (15.2 g of high purity, 49% yield, 3.5 g of low purity, 11% yield). 1H NMR (300.13 MHz, CDCl3) δ 4.86 (d, J = 63.5 Hz, 1H), 4.16-3.97 (m, 2.5H), 3.88 (d, J = 10.1 Hz, 0.5H), 3.15 (dd, J = 31.3, 9.8 Hz, 1H), 2.58 (d, J = 8.4 Hz, 1H), 2.35 (m, 2H), 2.18-1.48 (m, 8H), 1.44 (m, 9H), 1.40-1.30 (m, 4H), 0.87 (m, 6H) ppm. Scale-up batch: To a solution of I-04 crude (426.8 mmol) in DMF (500 mL), solid NaHCO3 (231.6 g, 2.76 mmol) and 3-(iodomethyl)pentane (159.7 g, 754.5 mmol) were added at room temperature under nitrogen atmosphere, and the mixture was stirred at 35-40 °C for 3 days. The reaction was monitored by HPLC, and after completion of the reaction, the mixture was filtered and the solid was washed with acetonitrile (1.5 L). The organic layers were combined and concentrated, and the crude residue obtained was redissolved in ethyl acetate (2 L). The solution was washed with water (500 mL), brine (500 mL), dried over sodium sulfate, and concentrated under vacuum to give the crude product I-05 as a red semi-solid (92.6 g, 57% yield) which was used directly without further purification.

[0125] Preparation of 2-tert-butyl 3-nonyl (3S,4aS,8aR)-6-oxo-decahydroisoquinoline-2,3-dicarboxylate (I-06) [ka] To a solution of I-04 (0.5 mmol) in DMF (2.0 mL) is added NaHCO3 (3.5 mmol) and 1-iodononane (1.3 mmol) at room temperature under nitrogen atmosphere, and the mixture is stirred at room temperature for 24 h. The reaction is monitored by HPLC, and after completion of the reaction, it is poured into water (30 mL) and extracted with ethyl acetate (2 x 30 mL). The combined organic layers are dried over sodium sulfate and concentrated under vacuum. The resulting residue is purified by flash column chromatography (silica gel, ethyl acetate / hexane) to give I-06.

[0126] Preparation of 3-(2R)-butano-2-yl 2-tert-butyl (3S,4aS,8aR)-6-oxo-decahydroisoquinoline-2,3-dicarboxylate (I-07) [ka] To a mixture of I-04 (0.5 mmol) and (2R)-butanol (0.6 mmol) in dichloromethane (1.0 mL) is added DCC (0.6 mmol) and DMAP catalyst under nitrogen atmosphere. The mixture is stirred at room temperature for 18 h and monitored by TLC or HPLC. Upon completion, acetonitrile (10 mL) is added and the mixture is stirred for 5-10 min. The solid precipitate is removed by filtration through a sintered glass funnel and the solid is washed with acetonitrile (10 mL). The filtrate is concentrated under vacuum and the residue is purified by flash column chromatography (silica gel, ethyl acetate / hexanes) to give I-07.

[0127] <Synthesis of olefin compound IX> Preparation of 2-benzyl 3-ethyl (3S,4aR,6E,8aR)-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethylidene]-decahydroisoquinoline-2,3-dicarboxylate (I-08) [ka] To a solution of triphenyl[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]phosphanium bromide salt (VIII, 9.09 g, 20.7 mmol) and ketone (I-02, 6.2 g, 17.25 mmol) in anhydrous DMF (50 mL) was added 2.0 M NaHMDS in THF (24.15 mL, 48.3 mmol) at 0°C to -10°C under nitrogen atmosphere. The internal temperature of the reaction was maintained at 0°C during the addition. The mixture was stirred at this temperature for 30 min and then allowed to warm to room temperature. After stirring at room temperature for 18 h, the mixture was slowly quenched with ice-cold brine solution (120 mL) and extracted with MTBE (7 x 250 mL) to partially remove triphenylphosphine oxide. The pH of the aqueous layer was adjusted to pH 2 using 3N HCl and extracted with ethyl acetate (4 x 250 mL). The combined organic layers were washed with water (2×200 mL), brine (200 mL), dried over sodium sulfate, and concentrated in vacuo. The residue was purified by flash column chromatography (silica gel, 10%→60% ethyl acetate / hexanes) to give I-08 as a pure white foamy solid (2.0 g, 26% yield) and a less pure yellow oil (2.0 g, 26% yield). 1H NMR (299.96 MHz, CDCl3) δ 7.38-7.27 (m, 5H), 5.50 (t, J = 6.7 Hz, 0.5H), 5.32 (dt, J = 21.5, 6.4 Hz, 0.5H), 5.23-5.14 (m, 1.5H), 5.06 (d, J = 12.9 Hz, 0.5H), 4.89 (d, J = 27.0 Hz, 1H), 4.17 (q, J = 6.8 Hz, 2H), 3.95-3.87 (m, 1H), 3.76-3.69 (m, 2H), 3.25 (d, J = 14.3 Hz, 0.5H), 3.15 (d, J = 12.9 Hz, 0.5H), 2.65 (d, J = 12.6 Hz, 0.5H), 2.47 (d, J = 13.8 Hz, 0.5H), 2.34 (d, J = 13.2 Hz, 0.5H), 2.24 (d, J = 11.4 Hz, 0.5H), 2.08-1.65 (m, 6H), 1.62-1.47 (m, 2H), 1.20 (d, J = 7.8 Hz, 3H) ppm.

[0128] Preparation of 2-tert-butyl 3-ethyl (3S,4aR,8aR)-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethylidene]-decahydroisoquinoline-2,3-dicarboxylate (I-09) [ka] To a solution of triphenyl[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]phosphanium bromide salt (VIII, 6.16 g, 14.0 mmol) and ketone (I-03, 3.8 g, 11.7 mmol) in anhydrous DMF (35 mL) was added 2.0 M NaHMDS in THF (16.4 mL, 32.8 mmol) at 0°C to -10°C under nitrogen atmosphere. The internal temperature of the reaction was maintained at 0°C during the addition. The mixture was stirred at this temperature for 30 min and then allowed to warm to room temperature. After stirring at room temperature for 18 h, the mixture was slowly quenched with ice-cold brine solution (50 mL) and extracted with MTBE (8 x 60 mL) to partially remove triphenylphosphine oxide. The pH of the aqueous layer was adjusted to pH 2 using 3N HCl and extracted with ethyl acetate (4 x 250 mL). The combined organic layers were washed with water (2×200 mL), brine (200 mL), dried over sodium sulfate, and concentrated in vacuo to give I-09 as a red oil (6.1 g, crude). 1 H NMR (300.13 MHz, CDCl3) δ 5.37 (m, 1H), 4.82 (dd, J = 48.5, 3.5 Hz, 0.5H), 4.77 (d, J = 45.5 Hz, 0.5H), 4.21-4.18 (m, 2H), 3.87-3.78 (m, 1H), 3.25-3.12 (m, 3H), 2.49-2.47 (m, 1.5H), 2.34 (d, J = 13.1 Hz, 0.5H), 2.1.0-1.60 (m, 8H), 1.54-1.46 (m, 9H), 1.30-1.22 (m, 3H) ppm.

[0129] Preparation of 2-tert-butyl 3-(2-ethylbutyl)(3S,4aR,8aR)-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethylidene]-decahydroisoquinoline-2,3-dicarboxylate (I-10) [ka] To a solution of the Wittig salt of tetrazole (VIII, 23.37 g, 53.2 mmol) and ketone (I-04, 16.9 g, 44.3 mmol) in anhydrous DMF (150 mL) was added 2.0 M NaHMDS in THF (62 mL, 124 mmol) at 0° C. to −10° C. under nitrogen atmosphere. The internal temperature of the reaction was maintained at 0° C. during the addition. The mixture was stirred at this temperature for 30 min and then allowed to warm to room temperature. After stirring at room temperature for 2 h, the mixture was slowly quenched with 10% aqueous brine solution (200 mL) and extracted with MTBE (4×300 mL) to partially remove triphenylphosphine oxide. The pH of the aqueous layer was adjusted to pH 2 using 3N HCl and extracted with ethyl acetate (4×300 mL). The combined organic layers were washed with water (2×150 mL), brine (200 mL), dried over sodium sulfate, and concentrated in vacuo. The residue was purified by flash column chromatography (silica gel, 0%→50% ethyl acetate / hexanes) to give I-10 as a foamy solid (13.1 g, 64% yield, high purity) and a pale yellow semi-solid (2.6 g, 13% yield, low purity). 1 H NMR (299.96 MHz, CDCl3) δ 5.55 (t, J = 7.0 Hz, 0.4H), 5.38 (dt, J = 20.3, 16.8 Hz, 0.6H), 4.84 (d, J = 46.8 Hz, 1H), 4.09-4.00 (m, 2H), 3.89-3.73 (m, 3H), 3.14-3.03 (m, 1H), 2.69 (t, J = 10.6 Hz, 0.6H), 2.49 (dd, J = 13.6, 6.1 Hz, 0.4H), 2.34 (d, J = 13.5 Hz, 0.6H), 2.31-2.21 (m, 0.4H), 2.09 (d, J = 14.7 Hz, 1H), 2.01 (t, J = 11.7 Hz, 1H), 1.90-1.75 (m, 4H), 1.65-1.40 (m, 3H), 1.45 (s, 9H), 1.40-1.26 (m, 5H), 0.91-0.85 (m, 6H) ppm.

[0130] Scale-up batch: To a solution of the Wittig salt of tetrazole (VIII, 121.8 g, 277.3 mmol) and crude ketone (I-04, 88 g, 230.7 mmol) in anhydrous DMF (670 mL) was added 2.0 M NaHMDS in THF (323.3 mL, 646.6 mmol) at 0°C to -10°C under nitrogen atmosphere. The internal temperature of the reaction was maintained at 0°C during the addition. The mixture was stirred at this temperature for 30 min and then allowed to warm to room temperature. After stirring at room temperature for 2 h, the mixture was slowly quenched with 10% aqueous brine solution (500 mL) and extracted with MTBE (2 x 400 mL) to partially remove triphenylphosphine oxide. The pH of the aqueous layer was adjusted to pH 2 using 3N HCl and extracted with ethyl acetate (4 x 1 L). The combined organic layers were washed with water (2×1 L), brine (500 mL), dried over sodium sulfate, and concentrated in vacuo. The residue was purified by flash column chromatography (silica gel, 0%→50% ethyl acetate / hexanes) to give I-10 as a foamy solid (51.6 g, 49% yield).

[0131] <Synthesis of fluorinated carbamate ester X> Preparation of 2-benzyl 3-ethyl (3S,4aS,8aR)-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-2,3-dicarboxylate (I-11) [ka] A 500 mL three-neck flask was prepared with a nitrogen inlet, a temperature probe, and a cooling bath. Water (85 mL) was placed in the flask, followed by iron(III) nitrate nonahydrate (Fe(NO3)3·9H2O (1.46 g, 3.04 mmol) and the mixture was stirred until dissolved (Solution A). Acetonitrile (85 mL) was placed in a 500 mL round bottom flask, followed by I-08 (667 mg, 1.517 mmol) and Selectfluor® (1.61 g, 4.55 mmol) and the mixture was stirred until dissolved (Solution B). Solution B was added to Solution A and stirred simultaneously at 22-25 °C. A clear yellow solution was observed. The pH of the solution was measured to be pH 2. The reaction was degassed using nitrogen sparging for 10 minutes and the mixture was cooled to -10°C. Sodium borohydride (591.2 mg) was charged in solid form in four portions over 5-10 minutes. The mixture was stirred at -10°C for 2 hours, then the mixture was warmed to 22-25°C and stirring was continued for 5 hours with HPLC monitoring. The mixture was concentrated under vacuum using a rotary evaporator to remove acetonitrile. 1N tetrahydrofuran was added to the mixture with stirring and maintaining the temperature below 25°C. HCl (110 mL) was added to adjust the pH to 2. The solution was extracted with ethyl acetate (4×100 mL). The organic layer was washed with water (2×100 mL) and brine (100 mL), and the solution was dried over sodium sulfate and concentrated under vacuum using a rotary evaporator to give the crude product (690 mg, 100%). The resulting residue was purified by flash column chromatography (silica gel, 0%→6% methanol / dichloromethane) to give I-11 as a white foamy solid (590 mg, 85% yield). 1H NMR (299.96 MHz, CDCl3) δ 7.35-7.26 (m, 5H), 5.25-5.09 (m, 2H), 4.93 (dd, J = 24.9, 5.4 Hz, 0.6H), 4.62 (m, 0.4H), 4.21-4.10 (m, 2H), 4.00 (t, J = 11.2 Hz, 0.6H), 3.61 (dd, J = 12.9, 6.6 Hz, 0.4H), 3.43 (dd, J = 13.5, 3.6 Hz, 0.4H), 3.27 (t, J = 14.5 Hz, 0.6H), 3.17-3.05 (m, 2H), 2.39-1.36 (m, 12H), 1.27-1.21 (m, 3H) ppm.

[0132] Preparation of 2-tert-butyl 3-ethyl (3S,4aS,8aR)-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-2,3-dicarboxylate (I-12) [ka] A 3 L three-neck flask was prepared with a nitrogen inlet, a temperature probe, and a cooling bath. Water (1.0 L) was placed in the flask followed by iron(III) nitrate nonahydrate (Fe(NO3)3·9H2O, 44.17 g, 109.34 mmol) and the mixture was stirred until dissolved (Solution A). Acetonitrile (1.0 L) was placed in a 2 L round bottom flask followed by I-09 (14.3 g, 35.27 mmol) and Selectfluor® (38.73 g, 109 mmol) and the mixture was stirred until dissolved (Solution B). Solution B was added to Solution A and stirred simultaneously at 22-25°C. A clear yellow solution was observed and the pH of the solution was measured to be pH 2. The reaction was degassed for 30 minutes using nitrogen bubbling and the mixture was cooled to -10°C. Sodium borohydride (13.34 g, 353 mmol) was charged as a solid in 10 portions over 10-15 min. The mixture was stirred at -10°C for 2 h, then the mixture was warmed to 22-25°C and stirring was continued for 5 h with HPLC monitoring. The mixture was concentrated under vacuum using a rotary evaporator to remove acetonitrile. To this mixture was added 1N HCl (500 mL) with stirring and maintaining the temperature below 25°C to adjust the mixture to pH 2. The solution was extracted with ethyl acetate (4 x 500 mL). The combined organic layers were washed with water (2 x 500 mL) and brine (100 mL), dried over sodium sulfate, and concentrated under vacuum using a rotary evaporator to give the crude product (13.5 g, 90%). The resulting residue was purified by flash column chromatography (silica gel, 0% to 70% ethyl acetate / heptane) to afford I-12 as a mixture of diastereomers as a white foamy solid (11.36 g, 76% yield). 1H NMR (299.962 MHz, CDCl3) δ 4.86 (dd, J = 51.3, 5.1 Hz, 0.5H), 4.49 (t, J = 5.1 Hz, 0.5H), 4.25-4.14 (m, 2H), 3.90 (t, J = 15.0 Hz, 0.5H), 3.47 (dd, J = 13.3, 7.3 Hz, 0.5H), 3.37 (dd, J = 13.2, 4.8 Hz, 0.5H), 3.22-3.14 (m, 2H), 3.14-3.04 (m, 0.5H), 2.34-1.53 ​​(m, 12H), 1.49-1.46 (m, 9H), 1.27 (t, J = 6.8 Hz, 3H) ppm.

[0133] Preparation of 2-tert-butyl 3-(2-ethylbutyl)(3S,4aS,8aR)-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-2,3-dicarboxylate (I-13) [ka] A 2 L three-neck flask was prepared with a nitrogen inlet, a temperature probe, and a cooling bath. Water (400 mL) was placed in the flask followed by iron(III) nitrate nonahydrate (Fe(NO3)3·9H2O, 34.2 g, 84.63 mmol) and the mixture was stirred until dissolved (Solution A). Acetonitrile (400 mL) was placed in a 1 L round bottom flask followed by I-10 (12.6 g, 27.3 mmol) and Selectfluor® (30 g, 84.63 mmol) and the mixture was stirred until dissolved (Solution B). Solution B was added to Solution A and stirred simultaneously at 22-25°C. A clear yellow solution was observed and the pH of the solution was measured to be pH 2. The reaction was degassed for 30 minutes using nitrogen bubbling and the mixture was cooled to -10°C. Sodium borohydride (10.33 g, 273 mmol) was charged as a solid in 10 portions over 10-15 min. The mixture was stirred at -10°C for 2 h, then the mixture was warmed to 22-25°C and stirring was continued for 5 h with HPLC monitoring. The mixture was concentrated under vacuum using a rotary evaporator to remove acetonitrile. To this mixture was added 1N HCl (~300 mL) while stirring and maintaining the temperature below 25°C to adjust the mixture to pH 2. The solution was extracted with ethyl acetate (3 x 350 mL). The combined organic layers were washed with water (400 mL) and brine (300 mL), dried over sodium sulfate, and concentrated under vacuum using a rotary evaporator to give the crude product. The resulting residue was purified by flash column chromatography (silica gel, 0%→50% ethyl acetate / heptane) to afford I-13 as a mixture of diastereomers as a white foamy solid (8.03 g, 61% yield). 1H NMR (299.96 MHz, CDCl3) δ 5.55 (t, J = 7.0 Hz, 0.4H), 5.38 (dt, J = 20.3, 16.8 Hz, 0.6H), 4.84 (d, J = 46.8 Hz, 1H), 4.09-4.00 (m, 2H), 3.89-3.73 (m, 3H), 3.14-3.03 (m, 1H), 2.69 (t, J = 10.6 Hz, 0.6H), 2.49 (dd, J = 13.6, 6.1 Hz, 0.4H), 2.34 (d, J = 13.5 Hz, 0.6H), 2.31-2.21 (m, 0.4H), 2.09 (d, J = 14.7 Hz, 1H), 2.01 (t, J = 11.7 Hz, 1H), 1.90-1.75 (m, 4H), 1.65-1.40 (m, 3H), 1.45 (s, 9H), 1.40-1.26 (m, 5H), 0.91-0.85 (m, 6H) ppm.

[0134] Scale-up batch: A 5 L three-neck flask was prepared with a nitrogen inlet, temperature probe, and cooling bath. Water (1.5 L) was placed in the flask followed by iron(III) nitrate nonahydrate (Fe(NO3)3·9H2O, 72.3 g, 178.96 mmol) and the mixture was stirred until dissolved (Solution A). Acetonitrile (1.5 L) was placed in a 3 L round bottom flask followed by I-10 (26.6 g, 57.9 mmol) and Selectfluor® (63.4 g, 178.96 mmol) and the mixture was stirred until dissolved (Solution B). Solution B was added to Solution A and stirred simultaneously at 22-25°C. A clear yellow solution was observed and the pH of the solution was measured to be pH 2. The reaction was degassed for 30 minutes using nitrogen sparging and the mixture was cooled to -10°C. Sodium borohydride (21.8 g, 576.2 mmol) was charged as a solid in 10 portions over 10-15 min. The mixture was stirred at -10°C for 2 h, then the mixture was warmed to 22-25°C and stirring was continued for 5 h with HPLC monitoring. The mixture was concentrated under vacuum using a rotary evaporator to remove acetonitrile. To this mixture was added 1N HCl (~640 mL) with stirring, maintaining the temperature below 25°C, to adjust the mixture to pH 2. The solution was extracted with ethyl acetate (3 x 500 mL). The combined organic layers were washed with water (500 mL) and brine (500 mL), dried over sodium sulfate, and concentrated under vacuum using a rotary evaporator to obtain the crude product. The resulting residue was purified by flash column chromatography (silica gel, 0%→50% ethyl acetate / heptane) to afford I-13 as a mixture of diastereomers as a white foamy solid (21.7 g, 61% yield).

[0135] Preparation of (3S,4aS,8aR)-2-[(benzyloxy)carbonyl]-6-fluoro-6-[2-(2H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-3-carboxylic acid (I-14) [ka] Compound I-11 (0.59 g, 1.28 mmol) was dissolved in 9:1 ethanol-water (2.0 mL) and 2N NaOH (5.3 mL) was added. The mixture was stirred at room temperature for 4 h. The reaction was diluted with water (50 mL) and extracted with MTBE (50 mL) to remove organic impurities. The aqueous layer was acidified with 6N HCl to pH ∼2. The mixture was extracted with ethyl acetate (3 x 50 mL) and washed with brine (50 mL). The combined organic layers were dried over sodium sulfate and concentrated under vacuum using a rotary evaporator to give the crude product I-14 as a white solid (435 mg, 79% yield). The crude product was used directly in the next step without further purification.

[0136] Preparation of (3S,4aS,8aR)-2-[(tert-butoxy)carbonyl]-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-3-carboxylic acid (I-15) [ka] Compound I-12 (185 mg, 0.435 mmol) was dissolved in 9:1 ethanol-water (1.0 mL) and 2N NaOH (2.5 mL) was added. The mixture was stirred at room temperature for 4 h. The reaction was diluted with water (50 mL) and extracted with MTBE (50 mL) to remove organic impurities. The aqueous layer was acidified with 6N HCl to pH ∼2. The mixture was extracted with ethyl acetate (3 x 50 mL) and washed with brine (50 mL). The combined organic layers were dried over sodium sulfate and concentrated under vacuum using a rotary evaporator to give the crude product (183 mg), which was purified by silica gel column chromatography (ISCO gradient system, 0% to 10% methanol-dichloromethane) to give I-15 as an oil (143 mg, 83% yield). This material was used directly in the subsequent step.

[0137] Preparation of 2-benzyl 3-cyclohexyl (3S,4aS,8aR)-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-2,3-dicarboxylate (I-16) [ka] Compound I-14 (51.5 mg, 0.12 mmol) and cyclohexanol (14.4 mg, 0.144 mmol) were dissolved in dichloromethane (1 mL). To the mixture was added diisopropylcarbodiimide (20 mg, 0.16 mmol) and 4-dimethylaminopyridine catalyst (5 mg, 0.04 mmol) under nitrogen atmosphere and the reaction was stirred at room temperature for 16-18 h. The reaction was monitored by HPLC and upon completion, acetonitrile (1 mL) was added and the mixture was stirred for 5-10 min. The solid precipitate was removed by filtration through a sintered glass funnel and the solid was washed with acetonitrile (5 mL). The filtrate was concentrated under vacuum and the residue was purified by silica gel flash column chromatography (0%→5% methanol / dichloromethane) to give I-16 as a colorless oil (40.4 mg, 66% yield). 1 H NMR (299.96 MHz, CDCl3) δ 7.35-7.27 (m, 5H), 5.23-5.07 (m, 2H), 4.95-4.50 (m, 2H), 3.97 (t, J = 12.3 Hz, 0.4H), 3.89-3.73 (m, 0.4H), 3.61 (dd, J = 13.6, 6.7 Hz, 0.4H), 3.40 (dd, J = 12.9, 4.2 Hz, 0.3H), 3.32-3.20 (m, 0.5H), 3.16-3.05 (m, 2H), 2.34-1.65 (m, 13H), 1.51-1.25 (m, 8H), 1.16 (d, J = 6.3 Hz, 1H) ppm.

[0138] Preparation of 2-benzyl 3-octyl (3S,4aS,8aR)-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-2,3-dicarboxylate (I-17) [ka] Compound I-14 (54.8 mg, 0.127 mmol) and 1-octanol (20 mg, 0.153 mmol) were dissolved in dichloromethane (1.0 mL). To the mixture, dicyclohexylcarbodiimide (35 mg, 0.168 mmol) and 4-dimethylaminopyridine catalyst (5 mg, 0.037 mmol) were added under nitrogen atmosphere and the reaction was stirred at room temperature for 16-18 h. The reaction was monitored by HPLC and upon completion, the solid precipitate was removed by filtration through a sintered glass funnel and the solid was washed with acetonitrile (5 mL). The filtrate was concentrated under vacuum and the residue was purified by silica gel flash column chromatography (0%→5% methanol / dichloromethane) to give I-17 as a colorless oil (40.2 mg, 58% yield). 1 H NMR (299.96 MHz, CDCl3) δ 7.40-7.25 (m, 5H), 5.25-5.10 (m, 2H), 4.94 (d, J = 25.5 Hz, 0.65H), 4.65 (s, 0.35H), 4.10-3.95 (m, 3H), 3.70-3.60 (m, 1H), 3.45-3.20 (m, 1H), 3.19-3.05 (m, 2H), 2.35-1.56 (m, 24H), 0.87 (m, 3H) ppm.

[0139] <Synthesis of fluorinated ester amine prodrug XI> Preparation of ethyl (3S,4aS,6S,8aR)-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-3-carboxylate (3) [ka] Compound I-12 (4.14 g, 9.73 mmol) was dissolved in 7:1 dioxane-anisole (57 mL, 0.17 M) and treated with 4N HCl / dioxane (10 equiv, 24 mL). After 1 h at room temperature, the reaction was checked by HPLC, which indicated that the reaction was not complete. Additional 4N HCl (5 equiv, 12 mL) was added and stirring was continued at room temperature. After 1 h, HPLC indicated that the reaction was not complete, so additional 4N HCl (5 equiv, 12 mL) was added. After 30 min, HPLC again indicated that the reaction was incomplete, so additional 4N HCl (2 equiv, 5 mL) was added. The reaction was complete after 4 h. Nitrogen gas was bubbled through the reaction to remove excess HCl from the solution, and then the solvent was partially evaporated under vacuum to half its volume. Hexane was added to precipitate the HCl salt, and the supernatant was discarded. The residue was triturated with dioxane (2 mL) and hexane (2 mL) was added to completely precipitate the HCl salt of the desired product. The residue was dried under vacuum to give crude product 3 (3.48 g, 99.3% yield) as a mixture of fluorinated diastereoisomers. The residue was purified by SFC chromatography (Analytical SFC method - Column: 4.6 x 100 mm, Chromegabond Ethyl Pyridine (ES Industries, West Berlin, NJ, USA); Solvent A: CO2, Solvent B: Methanol with 0.1% triethylamine; Gradient method: 5% to 65% B over 4 min, hold at 65% B for 1 min, return to initial conditions at 4 mL / min; System pressure: 125 bar; Column temperature: 40 °C; Sample diluent: Methanol; Retention time (3): 2.07 min; Retention time (C6-F-isomers): 1.06 min).Purification by preparative SFC method - column: 3.0 × 25.0 cm, 2-Ethylpyridine (Princeton Chromatography Inc., Princeton, NJ, USA); solvent A: CO2, solvent B: methanol with 0.5% triethylamine; isocratic method: 25% solvent B, 100 g / min; system pressure: 100 bar; column temperature: 25 °C; sample diluent: methanol with 0.5% triethylamine) gave 3 as a semi-solid (414 mg, isomeric purity 96.2%). This material was further purified by Gilson reverse phase chromatography (5% → 50% acetonitrile with 0.1% TFA in 0.1% TFA in water, then 5% → 95% acetonitrile in water) to give the pure compound for further testing. 1 H NMR (299.96 MHz, CD3OD) δ 4.31 (q, J = 7.1 Hz, 2H), 4.04 (dd, J = 12.9, 3.9 Hz, 1H), 3.20 (t, J = 13.0 Hz, 1H), 3.09 (dd, J = 12.7, 4.3 Hz, 1H), 3.00 (t, J = 8.1 Hz, 2H), 2.33-2.42 (m, 1H), 2.15-1.90 (m, 7H), 1.84-1.65 (m, 3H), 1.57-1.45 (m, 1H), 1.32 (t, J = 7.2 Hz, 3H) ppm. 19 F NMR (282.22 MHz, CD3OD) δ -160.70 (m, uncorrected, TFA reference -76.97) ppm. Mass Analysis (ES+) = 326.24 [M+H] (Formula: C 15 H 24 FN5O2, Exact Mass: 325.19).

[0140] Preparation of 2-ethylbutyl (3S,4aS,6S,8aR)-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-3-carboxylate (4)

change

[0141] Scale-up batch: To a solution of I-13 (21.7 g, 45.06 mmol) in anhydrous THF (15 mL), 4N HCl / dioxane (112.5 mL, 450.0 mmol) was added with stirring at room temperature. After stirring at room temperature for 4 h, HPLC showed the reaction was complete. Nitrogen gas was bubbled through the reaction mixture to remove excess HCl and the solvent was partially evaporated under vacuum to half the volume. The mixture was diluted with 1:1 MTBE-heptane (150 mL) which caused an oily layer to separate, which was triturated and the supernatant discarded. The oily residue was triturated again with 1:1 MTBE-heptane (150 mL) to further remove lipophilic impurities and the supernatant discarded. The oily residue was placed under vacuum to remove residual solvent to give the crude product as a white solid and a mixture of fluoride diastereomers which was further dried under high vacuum to constant weight (14 g crude, 75% yield). The residue was purified by SFC chromatography (Analytical SFC method - Column: 4.6 x 100 mm, Chiralpak IC SFC (Chiral Technologies, West Chester, PA, USA); Solvent A: CO2, Solvent B: Ethanol with 0.1% ammonium hydroxide; Gradient: 5% to 65% solvent B, 4 mL / min for 4 min; System pressure: 125 bar; Column temperature: 40 °C; Sample diluent: Ethanol; SFC retention time (4): 2.90 min; Preparative SFC method 1 - Column: 2.1 × 25.0 cm, Chiralpak IC (Chiral Technologies, West Chester, PA, USA); Solvent A: CO2, Solvent B: Ethanol with 0.25% ammonium hydroxide; Isocratic: 35% solvent B, 70 g / min; System pressure: 100 bar; Column temperature: 25 °C; Sample diluent: Ethanol with 0.25% ammonium hydroxide; Preparative SFC method 2 - Purification was performed by: column: 2.0 × 25.0 cm, PVA-Sil (YMC, Allentown, PA, USA); solvent A: CO2, solvent B: ethanol with 0.25% ammonium hydroxide; isocratic method: 50% solvent B, 80 g / min; system pressure: 100 bar; column temperature: 25 °C; sample diluent: ethanol with 0.25% ammonium hydroxide).The desired diastereomeric fraction was then further purified by reverse phase chromatography (column: 19×50 mm XBridge OBD Prep C18 5 μm, 5% acetonitrile-95% water with 0.1% ammonium hydroxide for 2 min, then 5%→95% acetonitrile-water with 0.1% ammonium hydroxide for 3 min; flow rate: 25 mL / min; column temperature: 40° C.; sample diluent: 2:1:1 ethanol:acetonitrile:water). The desired fraction was concentrated by rotary evaporation at 35° C. The dried material was redissolved in 1:1 acetonitrile:water and the solution was concentrated by rotary evaporation to remove acetonitrile, then frozen and lyophilized to give 4 as a white solid (2.42 g, 90% pure, 97.8% ee).

[0142] Preparation of 2-ethylbutyl (3S,4aS,6R,8aR)-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-3-carboxylate (5) [ka] SFC chromatography to give compound 4 also afforded C6-F-isomer 5 (SFC retention time: 3.04 min) as a viscous colorless oil (isomer purity 97.9%). This material was further purified by Gilson reverse phase chromatography (15%→60% acetonitrile with 0.1% TFA-0.1% TFA in water, then 10%→98% acetonitrile-water) to give the pure compound for further testing. 1 H NMR (299.96 MHz, CD3OD) δ 4.30-4.25 (m, 2H), 4.18 (dd, J = 10.9, 5.5 Hz, 1H), 3.35-3.27 (m, 2H), 3.09 (t, J = 8.1 Hz, 2H), 2.28-1.94 (m, 7H), 1.86-1.69 (m, 5H), 1.59 (p, J = 6.1 Hz, 1H), 1.46-1.36 (m, 4H), 0.94 (t, J = 7.3 Hz, 6H) ppm.

[0143] Preparation of cyclohexyl (3S,4aS,6S,8aR)-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-3-carboxylate (6) [ka] Compound I-16 (100 mg, 0.2 mmol) was dissolved in THF (20 mL) and 10% Pd / C (100 mg) was added. The mixture was degassed by three hydrogen purge cycles (vacuum, open, and pressurized with hydrogen to 52 psi). The mixture was stirred under hydrogen overnight and then monitored by HPLC. The mixture was filtered through Celite®, washed with THF (20 mL), and the filtrate was concentrated under vacuum. The residue was dissolved in THF (20 mL), and fresh 10% Pd / C catalyst (100 mg) was added, followed by a hydrogen purge cycle as before. The mixture was stirred under hydrogen overnight and monitored by HPLC. The mixture was filtered through Celite®, washed with THF (100 mL), and the filtrate was concentrated under vacuum. The crude product was purified by Gilson reverse phase chromatography (20%→60% acetonitrile with 0.1% TFA-0.1% TFA in water) to give 6 as a TFA salt (17.0 mg, second peak) and the C6-F-isomer TFA salt (21.7 mg, first peak). The desired isomer was further purified by Gilson reverse phase chromatography (10%→98% acetonitrile-water) to give pure compound 6 for further testing. 1 H NMR (299.96 MHz, CD3OD) δ 4.89-4.94 (m, 1H), 4.01 (dd, J = 12.9, 3.9 Hz, 1H), 3.17 (t, J = 12.7 Hz, 1H), 3.06 (dd, J = 13.0, 4.6 Hz, 1H), 2.98 (t, J = 7.9 Hz, 2H), 2.33-2.42 (m, 1H), 1.97-2.15 (m, 5H), 1.83-1.96 (m, 3H), 1.64-1.82 (m, 5H), 1.25-1.62 (m, 8H) ppm.19 F NMR (282.22 MHz, CD3OD) δ -160.20 (m, uncorrected, TFA reference -76.97) ppm. Mass Analysis (ES+) = 380.19 [M+H] (Formula: C 19 H 30 FN5O2, Exact Mass: 379.24).

[0144] Preparation of octyl (3S,4aS,6S,8aR)-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-3-carboxylate (7) [ka] Compound I-17 (40 mg, 0.074 mmol) was dissolved in THF (10 mL) and 10% Pd / C (100 mg) was added. The mixture was degassed by three hydrogen purge cycles (vacuum, open, and pressurized with hydrogen to 52 psi). The mixture was stirred under hydrogen overnight and then monitored by HPLC. The mixture was filtered through Celite®, washed with THF (20 mL), and the filtrate was concentrated under vacuum. The residue was dissolved in THF (20 mL), and fresh 10% Pd / C catalyst (100 mg) was added, followed by hydrogen purge cycles as before. The mixture was stirred under hydrogen overnight and monitored by HPLC. The mixture was filtered through Celite®, washed with THF (100 mL), and the filtrate was concentrated under vacuum. The crude product was purified by Gilson (30%→60% acetonitrile containing 0.1% TFA-0.1% TFA in water) to give 7 as a TFA salt (2.9 mg, second peak) and the C6-F-isomer TFA salt (4.5 mg, first peak). 1H NMR (299.96 MHz, CD3OD) δ 4.26 (dt, J = 6.6, 1.8 Hz, 2H), 4.10 (dd, J = 12.7, 4.0 Hz, 1H), 3.23 (d, J = 13.2 Hz, 1H), 3.17-3.09 (m, 3H), 2.43-2.36 (m, 1H), 2.20-1.93 (m, 1H), 1.89-1.52 (m, 1H), 1.45-1.30 (m, 1H), 0.91 (t, J = 6.9 Hz, 3H) ppm. Mass Analysis (ES+) = 410.17 [M+H] (Formula: C 21 H 36 FN5O2, Exact Mass: 409.29).

[0145] Using the above protocol, additional ketone carbamate esters IV can be made, which can then be reacted with Wittig tetrazoles VIII, fluorinated, and deprotected to generate novel fluorinated ester amine prodrugs (XI) as shown in Table 1. [Table 1: Fluorinated ester amine prodrugs (XI)] [Table 1] TIFF0007653184000041.tif254121 TIFF0007653184000042.tif254121 TIFF0007653184000043.tif187170

[0146] <Synthesis of Fluorinated Decahydroisoquinoline Amino Acid XII> Preparation of (3S,4aS,6S,8aR)-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-3-carboxylic acid (1) [ka] To a solution of compound 3 (1.29 g, 3.97 mmol) in water (2 mL) was added 2N NaOH (9 mL, 16.9 mmol) at room temperature under nitrogen atmosphere. The solution was stirred at room temperature for 12 h and then quenched with 1N HCl (18.5 mL) and pH adjusted to pH 2-3. The compound was purified by a resin catch and release method, Dowex® 50WX8 Purification was performed with 200-400 mesh resin (Sigma-Aldrich, 1.7 equiv / mL swell, 14 mL) which was washed with water in a coarse pore sintered glass funnel until the pH was neutral. Resin (5 mL) was added to the compound solution and the mixture was stirred at room temperature for 10 min. Fresh resin (9 mL) was placed in a column equipped with a coarse frit and the mixture was decanted into the column (including all the resin). The top of the resin column was covered with cotton to prevent disturbance of the layers. The solution was allowed to elute slowly and naturally. The resin was then washed with water (3×30 mL) to remove salts, 1:1 THF:water (3×30 mL) to remove organic impurities, water (2×30 mL), and then 6% ammonium hydroxide (300 mL) to release the desired compound. Fractions were collected and the desired compound was detected using ninhydrin staining of reverse phase TLC plates (10% acetonitrile-water). The desired fractions were combined and concentrated under vacuum to give the crude product. The crude product was dissolved in water and lyophilized to give 1 as a white solid (1.02 g, 86% yield). 1 H NMR (299.96 MHz, D2O) δ 3.58 (dd, J = 12.9, 3.9 Hz, 1H), 3.11 (dd, J = 19.2, 12.9 Hz, 1H), 3.08 (s, 1H), 2.97 (dd, J = 8.8, 7.0 Hz, 2H), 2.35-2.28 (m, 1H), 2.14-1.61 (m, 9H), 1.56-1.47 (m, 1H), 1.38 (dtd, J = 42.6, 14.6, 4.6 Hz, 1H) ppm. 19F NMR (282.22 MHz, D2O) δ -157.59 (m, uncorrected) ppm. Mass Analysis (ES+) = 298.12 [M+H] (Formula: C 13 H 20 FN5O2, Exact Mass: 297.16).

[0147] Preparation of (3S,4aS,6R,8aR)-6-fluoro-6-[2-(1H-1,2,3,4-tetrazol-5-yl)ethyl]-decahydroisoquinoline-3-carboxylic acid (2) [ka] Using a similar protocol to make compound 1, the C6-F-isomer compound 5 was hydrolyzed with NaOH and purified by a resin catch-and-release method to give 2. 1 H NMR (299.96 MHz, D2O) δ 3.75 (dd, J = 10.6, 3.4 Hz, 1H), 3.29-3.14 (m, 2H), 2.96 (t, J = 7.8 Hz, 2H), 2.22 (dt, J = 7.8, 3.6 Hz, 1H), 2.13 (dt, J = 7.8, 3.2 Hz, 1H), 2.10-1.86 (m, 3H), 1.84-1.61 (m, 5H) ppm. 19 F NMR (282.22 MHz, D2O) δ -167.27 (uncorrected, TFA reference -75.69) ppm. Mass Analysis (ES+) = 298.26 [M+H] (Formula: C 13 H 20 FN5O2, Exact Mass: 297.16).

[0148] III. Materials, Methods and Experimental Data for In Vitro Biology and In Vivo Pharmacology The present disclosure relates to the chemical composition of substances (molecules) and characterizes the biological and pharmacological activity of the molecules as AMPA receptor (AMPAR) antagonists or as prodrugs of such molecules. The present disclosure further discloses applications for the treatment of pain, convulsions, seizures, epilepsy, and status epilepticus.

[0149] In addition, the biological activity of the compounds as antagonists of AMPA or NMDA receptors (also called ionotropic glutamate-gated ion channels) was carried out using ex vivo functional electrophysiological assays of pyramidal neurons in slices of the prefrontal cortex of rat brain. The effects of the test compounds compared to reference compounds were studied using whole-cell patch-clamp electrophysiological recordings of s-AMPA or NMDA induced currents, respectively, in prefrontal cortex (layer V) pyramidal neurons in rat brain slices.

[0150] <Brain slice preparation protocol> Male Sprague-Dawley rats were obtained from Charles River Laboratories (Wilmington, MA, USA) and housed four per cage in a controlled temperature (20.5–23.5°C) and humidity (30–80%) environment with a 12-h light / dark cycle and free access to food (Teklad Global Soy Protein, Cat. No. T.2920X10, Envigo, Indianapolis, IN, USA) and water. At 4–6 weeks of age, rats were terminally anesthetized using isofluorane [(1-chloro-2,2,2-trifluoroethyl difluoromethyl ether), supplied by Baxter Healthcare Corp, Deerfield, IL, USA] and decapitated. Brains were removed and 300 μm thick coronal prefrontal cortex (PFC) or sagittal hippocampal slices were cut using a vibratome microtome. After brain removal and throughout sectioning, tissues were submerged in ice-cold aqueous cerebrospinal fluid (aCSF). Once cut, slices were transferred to a beaker containing aCSF and allowed to sit at room temperature for a minimum of 1 h before electrophysiological recordings began. After this time, individual slices were transferred to a recording chamber that was continuously perfused with aCSF at a rate of 4–6 mL / min before the start of the experimental protocol. aCSF composition (in mM): NaCl, 127; KCl, 1.9; KH2PO4, 1.2; CaCl2, 2.4; MgCl2, 1.3; NaHCO3, 26; D-glucose, 10 (reagent suppliers listed below) in water equilibrated with 95% oxygen gas-5% CO2 gas. Experiments examining NMDA currents used aCSF supplemented with 10 μM glycine (reagent supplier listed below). All experiments were performed following an Institutional Animal Care and Use Committee (IACUC) approved protocol.

[0151] <Reagents for electrophysiological research> Fisher Scientific (Fair Lawn, NJ, USA) supplied NaCl product #S271, KCl product #P330, CaCl2 product #C79, MgCl2 product #M33, D-glucose product #D16, HEPES product #BP310, sucrose product #S5, and NaHCO3 product #S233. Millipore-Sigma (St. Louis, MO, USA) supplied Mg-ATP product #A9187, CsCl product #C3032, EGTA-Na product #E4378, GTP product #G8877, glycine product #G7126, KOH product #417661, and D-potassium gluconate product #G4500. EMD Chemicals (Gibbstown, NJ, USA) supplied KH2PO4 product #PX1565. Tocris and BioTechne (Bristol, UK and Minneapolis, MN, USA) supplied (S)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (s-AMPA), product #0254, and N-methyl-D-aspartic acid (NMDA), product #0114. Thermo Scientific (Rockford, IL, USA) supplied DMSO product #20688.

[0152] <Electrophysiological recordings of s-AMPA or NMDA-induced currents in pyramidal neurons of rat prefrontal cortical brain slices> Whole-cell patch-clamp recordings were performed on layer V prefrontal cortical pyramidal neurons at room temperature using a "visualization" version of the patch-clamp technique. Neurons were visualized using a BX51 upright microscope (Olympus, Richmond Hill, Ontario, Canada) fitted with a 40x LUMPlanFl water-immersion objective. The microscope was connected to a C2400 CCD camera (Hamamatsu, Bridgewater, NJ, USA) and images were viewed on a VM5516 B / W monitor (Sanyo, Moriguchi, Osaka, Japan). Electrophysiological recordings were obtained using a Multiclamp™ 700B patch-clamp amplifier (Molecular Devices, Sunnyvale, CA, USA) with analog signals digitized with a Digidata® 1440a (Molecular Devices, Sunnyvale, CA, USA). Patch pipettes were made from thin-walled borosilicate glass, type GC150TF-10 (Harvard Apparatus, Saint-Laurent, Quebec, Canada), with resistances of 3–8 MΩ when filled with intracellular solution, using a P-87 Flaming / Brown micropipette puller (Sutter, Novato, CA, USA).

[0153] The composition of the intracellular solutions used for recording from PFC neurons was as follows (mM): potassium d-gluconate, 140; KCl, 10; EGTA-Na, 1; HEPES, 10; Mg-ATP, 4, 0.3 GTP; pH and osmolality in all intracellular solutions were corrected with potassium hydroxide and sucrose, respectively (reagent suppliers listed above). All test compound experiments were performed in the voltage clamp recording configuration of the whole-cell patch clamp technique, with all recordings made at a holding potential of -60 mV. Recordings were monitored and digitized at 10 kHz on a Dell personal computer (PC) running Axon™ pClamp™ software (Molecular Devices, Sunnyvale, CA, USA).

[0154] <AMPA receptor biological activity assay of compounds in neurons from brain slices> Compounds were tested in whole-cell patch-clamp electrophysiology experiments to measure antagonistic effects on s-AMPA-induced currents. In these experiments, the effect of a single concentration of each test compound on currents evoked by 20 μM s-AMPA in layer V rat brain slice prefrontal cortical pyramidal neurons was investigated. Both 1 μM and 10 μM concentrations of compound 1, as well as 1 μM concentration of compound 2, were tested in separate experiments on brain slices, with the baths perfused from a 50 mL syringe configured in series with the main perfusion line from an aCSF reservoir. 20 μM s-AMPA was pressure-injected for 100–1000 ms every 1–2 min using a pneumatic picopump, model NPI PDES-02DX (npi Electronic GmbH, Tamm, Germany), connected directly to a microelectrode placed 200 μm inside the recording neuron. Each protocol was repeated three times. If the peak current was reduced by more than 70% upon the first application of 10 μM of the test compound, one experiment was performed at this 10 μM compound concentration, in which case the compound was tested three times at 1 μM concentration following a single test at 10 μM.

[0155] <NMDA receptor biological activity assay of compounds in neurons from brain slices> Compounds are tested in whole-cell patch clamp electrophysiology experiments to measure antagonist effects on NMDA-induced currents. For NMDA-induced current experiments, the effect of a single concentration of each test compound on currents evoked by 50 μM NMDA in layer V rat brain slice prefrontal cortical pyramidal neurons was investigated. Compound 1 at a concentration of 30 μM was administered to the slices by bath perfusion from a 50 mL syringe configured in series with the main perfusion line from an aCSF reservoir. 50 μM NMDA was pressure-injected for 100-1000 ms every 1-2 min using a pneumatic picopump, NPI PDES-02DX type (NPI Electronics GmbH, Tamm, Germany) connected directly to a microelectrode placed 200 μm inside the neuron to be recorded. Compound 1 was tested in triplicate at a concentration of 30 μM. Preparations of test or reference compounds, or inducer s-AMPA or inducer NMDA. Test compounds (Sea Pharmaceuticals LLC, Cambridge, MA, USA) were prepared as 10 mM or 30 mM stock solutions in 100% DMSO solvent (solvent supplier listed above). Current inducers s-AMPA or NMDA were made up as 20 μM or 50 μM stock solutions in aCSF, respectively. Test compound stock solutions were diluted to the indicated final test concentrations in the appropriate external recording solution immediately prior to use. All compounds were stored at -20°C prior to use.

[0156] <Testing of Reference Compounds> The Texan panel was tested as a reference compound in the s-AMPA electrophysiological assay at concentrations of 30 μM, 10 μM, 3 μM, 1 μM, 0.3 μM, and 0.1 μM. The compounds concentration-dependently inhibited the peak amplitude response of rat brain slice PFC pyramidal neurons to 20 μM s-AMPA (IC 50 The observed concentration was 481 ± 84 nM in s-AMPA-induced currents in rat cortical pyramidal neurons (n ​​= 4–5 experiments per concentration). Tezane panels were tested in electrophysiological assays of NMDA at a concentration of 30 μM, which partially inhibited (40% ± 2%, n = 4 experiments) the peak amplitude response of rat brain slice PFC pyramidal neurons to 50 μM NMDA-induced currents in rat cortical pyramidal neurons.

[0157] <Electrophysiological Experimental Data and Statistical Analysis>

[0158] All data were sampled using pClamp Clampex acquisition software and all offline analyses were performed using Clampfit (MDS Analytical Technologies). Data editing and plotting were performed using Excel® (Microsoft). One-way repeated measures analysis of variance (ANOVA, Graphpad Prism) with Dunnett's post-hoc comparisons was used for statistical analysis. Table 2: Summary of in vitro biological data for Compound #1 and Compound #2. [Table 2]

[0159] In vivo pharmacology of compounds following treatment in rodents, including in vivo rodent models of epilepsy or epileptic seizures and in vivo rodent models of pain.

[0160] Formulation and preparation of compounds for administration to animals Dosing suspensions or solutions of test compounds were prepared as specified in one of two formulations: (i) a DMSO in aqueous methylcellulose formulation (referred to as DMC) or (ii) a pH-adjusted saline formulation (referred to as SPHA). For the DMC formulations, test compounds 3, 4, or 6 were dissolved in DMSO and diluted in 0.5% methylcellulose (with a final DMSO concentration of 3% or less). 0.5% methylcellulose (0.5% MC, Sigma Cat. No. M-0430, St. Louis, MO, USA) was prepared in water as per the manufacturer's instructions. The DMC mixture of compounds was homogenized by vortexing and stirred on a hot plate (~40°C) until the test compound mixture was a homogenous suspension or completely dissolved. The SPHA formulation was used for some test articles that were more water-soluble compounds or salts. For the SPHA formulations, compound powder was mixed directly into physiological (normal) saline (0.9% NaCl), then 0.1N or 1N sodium hydroxide solution was carefully added to a pH of 9-9.5, and the sample was stirred and vortexed while heating at approximately 40°C until complete dissolution (within 15 minutes). 0.1N or 1N hydrochloric acid solution was then carefully added to adjust the pH to pH 7.1-7.3. Alternatively, 0.5% methylcellulose in water could be used instead of normal saline. Vehicle solutions were prepared using the same protocol above without compound. Dosing mixtures were allowed to come to room temperature prior to administration. Dosing mixtures were prepared fresh on the day of testing and used within 3 hours. All dosing solutions or suspensions were thoroughly mixed prior to administration.

[0161] Compound #1 was formulated in SPHA. Other carboxylic acid isomers and their salts can be similarly formulated. Dosing solutions of levetiracetam (TCI America, Portland, OR, USA) were prepared in 0.5% methylcellulose (MC) in water.

[0162] The compounds may also be formulated in other vehicles, including but not limited to those mentioned above.

[0163] Animal handling protocols and administration of compounds or vehicle in rodent seizure models (mouse or rat) Adult male Carworth Farms (CF-1) mice (25–35 g) or juvenile male Sprague Dawley (SD) rats (100–150 g) were obtained from Charles River Laboratories, Inc. (Wilmington, MA, USA). CF1 mice were typically used for the 6 Hz psychomotor seizure model (6 Hz model described in the following section) but could also be used for the maximal electroshock seizure (MES) model when performed in mice (maximal electroshock seizure model described in the following section). SD rats were typically used for the MES model but could also be used for the 6 Hz psychomotor model when performed in rats. Animals had free access to food and water except during the study period. After delivery from the supplier's laboratory to the in vivo pharmacology laboratory, animals were allowed ample time to acclimate to the housing conditions before testing. Animals were housed in plastic cages in a room with controlled humidity, ventilation, and lighting (12 h light and 12 h dark). Animals were maintained in accordance with the recommendations of the Guide for Care and Use of Laboratory Animals (National Research Council) and in accordance with the guidelines established by the Institutional Animal Care and Use Committee (IACUC). Animal experiments were performed in accordance with the Animal Research: Reporting of In Vivo Experiments (ARRIVE) guidelines (UK) and approved by the IACUC. Test compounds or their respective vehicles (placebo) were administered using optimal fluid volume to body fluid ratios. Solutions or suspensions of test compounds, reference compounds or vehicles were administered to mice or rats at 0.01 mL / g (mice) or 0.004 mL / g (rats) of body weight by subcutaneous (sc) injection, intraperitoneal (ip) injection or by oral gavage (po) unless otherwise specified. The reference drug levetiracetam or vehicle was administered by intraperitoneal (ip) injection.

[0164] <Description of the literature and in vivo pharmacological validation of the 6 Hertz (6 Hz) psychomotor seizure model in rodents> Dr. James EP Toman first described the 6 Hz psychomotor seizure model in 1951. The 6 Hz psychomotor seizure model was extensively characterized in 1953 by pharmacologist Louis S. Goodman, who showed that certain clinical AEDs were ineffective in the mouse 6 Hz model versus the mouse MES model (described below in another section), and was pharmacologically tested in mice using clinical antiepileptic drugs (AEDs) and was found to be resistant to treatment with certain AEDs such as phenytoin. The mouse 6 Hz psychomotor seizure model was little used for the next 50 years until it was reviewed in 2001 by H. Steven White and Harold H. Wolf, who compared the pharmacology of several types of AEDs. Toman JE, Neurology, 1951, Vol, 1, p.444-460. Brown WC et al., J Pharmacol Exp Ther, (USA), 1953, Vol. 107, p.273-283. Barton ME et al., Epilepsy Res, 2001, Vol. 47, p.217-227. Metcalf CS et al., Epilepsia, 2017a, Vol. 58, p.484-493. Metcalf CS, Epilepsia, 2017b, Vol. 58, p.1073-1084.

[0165] 6Hz Psychomotor Seizure Test for Antiseizure Activity of Compounds After topical anesthesia of the cornea of ​​each mouse eye, corneal electrodes are placed. 6 Hz psychomotor seizures are induced in mice (usually 8 per group) using electrical stimulation via corneal electrodes (6 Hz, 0.2 ms rectangular pulse, 22 mA lasting 3 seconds, using a Grass 48 stimulator as described in Barton ME et al., Epilepsy Res, 2001, Vol. 47, p.217-227). Prior to placement of the corneal electrodes for electrical stimulation, drops of 0.5% tetracaine saline solution (Sigma) were applied to each eye. Seizures and behaviors resulting from electrical stimulation in the 6 Hz model include a minor clonic phase followed by stereotypical automatisms such as syncope, forelimb clonus, vibrissa twitching, and Straub's tail reaction. Animals were observed by a pharmacologist for these behaviors for 1 minute after stimulation. If any of these behaviors were observed, the animal was considered to have exhibited seizures. Animals that did not exhibit any of these behaviors were considered "protected" from seizures. This serves as a screening test for the in vivo pharmacological activity of test substances or reference antiepileptic drugs and defines antiseizure activity as shown in several publications [Barton ME et al., Epilepsy Res., 2001, Vol. 47, p.217-227; Barton ME et al., Epilepsy Res., 2003, Vol. 56, p.17-26; Brown WC et al., J Pharmacol Exp Ther, 1953, Vol. 107, p.273-283; Metcalf CS et al., Epilepsia, 2017a, Vol. 58, p.484-493; Metcalf CS et al., Epilepsia, 2017c, Vol. 58, p.239-246.].

[0166] Unless otherwise specified, the pretreatment time for mice or rats was typically 0.5 hours for the respective vehicle, compounds 1, 3, 4, 6 or other compounds administered by subcutaneous or intraperitoneal route (unless other times are indicated). For compounds 3, 4, 6 or other compounds administered by oral route, a pretreatment time of 1 hour was used unless other times are indicated. Test compounds or respective vehicles were occasionally tested at other pretreatment times as indicated. Median effective dose (ED 50 ) and 95% confidence intervals (CI) were calculated using Prism (Graphpad software).

[0167] <References on the 6 Hz psychomotor seizure model in rodents> Toman JE 1951. Neurology 1:444-460. "Neuropharmacologic considerations in psychic seizures." Brown WC, Schiffman DO, Swinyard EA, Goodman LS. 1953. J Pharmacol Exp Ther 107:273-283. "Comparative Assay of an Antiepileptic Drugs by Psychomotor Seizure Test and Minimal Electroshock Threshold Test." Barton ME, Klein BD, Wolf HH, White HS. 2001. Epilepsy Res 47:217-227. "Pharmacological characterization of the 6 Hz psychomotor seizure model of partial epilepsy." Metcalf CS, Klein BD, Smith MD, Pruess T, Ceusters M, Lavreysen H, Pype S, Van Osselaer N, Twyman R, White HS. 2017a. Epilepsia 58:484-493. Efficacy of mGlu2 positive allosteric modulators alone and in combination with levetiracetam in the mouse 6 Hz model of psychomotor seizures; Metcalf CS, West PJ, Thomson KE, Edwards SF, Smith MD, White HS, Wilcox KS. 2017b. Epilepsia 58:1073-1084. "Development and pharmacologic characterization of the rat 6 Hz model of partial seizures." Metcalf CS, Klein BD, McDougle DR, Zhang L, Kaufmann D, Grzegorz Bulaj G, White HS. 2017c. Epilepsia 58:239-246. "Preclinical Evaluation of Intravenous NAX 810-2, a Novel GalR2-preferring Analog, for Anticonvulsant Efficacy and Pharmacokinetics."

[0168] Literature description of the MES (maximal electroshock seizure) model for testing molecules for anticonvulsant, antiseizure and antiepileptic activity in rodents Pharmacologist Louis S. Goodman extensively characterized the in vivo pharmacology of the MES model in therapeutic studies comparing experimental anticonvulsant, antiseizure, and antiepileptic agents with antiepileptic drugs (AEDs) in the 1950s–1970s. Subsequent work by H. Steve White and Harold H. Wolf in the 1980s–2010s further expanded the use of the MES seizure model for pharmacological characterization of antiepileptic compounds.

[0169] Materials, methods, and validation of several clinical antiepileptic drugs in the MES model are described in Swinyard EA et al., J Pharmacol Exp Ther, (USA), 1952, Vol. 106, p.319-330. The method for Goodman's MES model used an electrical stimulation device and electrodes described in Woodbury LA, Davenport VD., Arch Int Pharmacodyn Ther, 1952, Vol. 92, p.97-107. MES animal models and this type of MES device have been used extensively by the National Institutes of Health in the past decades to pharmacologically characterize the efficacy of anticonvulsant agents and antiepileptic drugs (AEDs) in vivo when screening compounds for anticonvulsant activity (the first two references are chapters in two books). White HS et al. (Levy RH, Mattson RH, Meldrum BS, eds. "Antiepileptic Drugs," 4th ed., 1995, pp. 99-110);White HS et al. (Levy R, Mattson R, Meldrum B, Perucca E, eds. "Antiepileptic Drugs," 5th ed., 2002, pp. 36-48);White HS et al., Italian Journal Neurological Sciences, 1995, Vol. 16, p. 73-77;White HS et al., Advances in Neurol, 1998, Vol. 76, p. 29-39. (Review);White HS et al., Epilepsia, 2008, Vol. 49, p. 1213-1220. (Methods: Description of the revised MES model);Barton ME, Peters SC, Shannon HE, Epilepsy Res., 2003, Vol. 56, p.17-26 (detailed methods for the mouse 6Hz model and mouse MES model).

[0170] <References for the rodent maximal electroshock seizure (MES) model in rats or mice> Swinyard EA, Brown WC, Goodman LS. 1952. J Pharmacol Exp Ther 106:319-330. "Comparative assays of antiepileptic drugs in mice and rats" Woodbury LA, Davenport VD. 1952. Arch Int Pharmacodyn Ther 92:97-107. "Design and use of a new electroshock seizure apparatus, and analysis of factors altering seizure threshold and pattern." White HS, Woodhead JH, Franklin MR. Swinyard EA, Wolf HH. (1995) General principles: experimental selection, quantification, and evaluation of antiepileptic drugs. In Levy RH, Mattson RH, Meldrum BS (Eds) Book title: Antiepileptic Drugs . 4th edition. Raven, New York, pp. 99-110; White HS, Woodhead JH, Wilcox KS, Stables JP, Kupferberg HJ, Wolf HH. (2002) Discovery and preclinical development of antiepileptic drugs. In Levy R,Mattson R, Meldrum B, Perucca E (Eds) Book title: Antiepileptic Drugs . 5th edition. Lippincott Williams & Wilkins, Philadelphia, pp. 36-48; White HS, Johnson M, Wolf HH, Kupferberg HJ. 1995. Italian Journal Neurological Sciences 16:73-77. "The early identification of anticonvulsant activity: role of the maximal electroshock and subcutaneous pentylenetetrazol seizure models." (Review); White HS, Wolf HH, Woodhead JH, Kupferberg HJ. 1998. Advances in Neurol 76:29-39. "The national institutes of health anticonvulsant drug development program: screening for efficacy." (Review); White HS, Franklin MR, Kupferberg HJ, Schmutz M, Stables JP, Wolf HH. 2008. Epilepsia 49:1213-1220. "The anticonvulsant profile of rufinamide (CGP 33101) in rodent seizure models." Leander JD, Rathbun RC, Zimmerman DM. 1988. Brain Res. 454:368-732"Anticonvulsant effects of phencyclidine-like drugs: relation to N-methyl-D-aspartic acid antagonism"; Leander JD. 1989. Epilepsy Res. 4: 28-33 "Evaluation of dextromethorphan and carbetapentane as anti-convulsants and N-methyl-D-aspartic acid antagonists in mice"; Yamaguchi S, Donevan SD, Rogawski MA. 1993. Epilepsy Res.15:179-184 "Anticonvulsant activity of AMPA / kainate antagonists: comparison of GYKI 52466 and NBOX in maximal electroshock and chemoconvulsant seizure models." Barton ME, Peters SC, Shannon HE. 2003 Epilepsy Res. 56:17-26 "Comparison of the effect of glutamate receptor modulators in the 6 Hz and maximal electroshock seizure models Metcalf CS, West PJ, Thomson KE, Edwards SF, Smith MD, White HS, Wilcox KS. 2017c. Epilepsia 58:1073-1084. "Development and pharmacologic characterization of the rat 6 Hz model of partial seizures".

[0171] <In Vivo Pharmacological Test of the Maximal Electroshock Seizure (MES) Model of Compounds Related to Anticonvulsant Activity in Rats> Male Sprague-Dawley rats (body weight 100-150 g at the time of testing) were obtained from Charles River Laboratories (Wilmington, MA, USA) (N=8-10 per group). Rats were administered test substances (experimental compounds, or vehicle, or positive control reference anticonvulsant compounds) by intraperitoneal (ip), subcutaneous (sc), intravenous (iv), or oral gavage (po) routes. At the time of dosing and again before corneal stimulation, rats were corneal anesthetized with 0.5% tetracaine saline solution. Unless otherwise specified, pretreatment time for Compound 1 was 30 min before corneal stimulation with electrodes (electrical stimulation AC current 60 Hz, 150 mA, duration 0.2 s for rats) (pretreatment time for vehicle or other test compounds was 60 min or as indicated). When mice are used, stimulation parameters are 60 Hz, 50 mA alternating current for 0.2 seconds, following published methods [Metcalf CS et al., Epilepsia, 2017c, Vol. 58, p.1073-1084;White HS et al., Italian Journal Neurological Sciences, 1995, Vol. 16, p.73-77;Barton ME et al., Epilepsy Res., 2003, Vol. 56, p.17-26 (both of which contain detailed descriptions of the mouse MES model and the 6 Hz model);Leander JD, Rathbun RC, Zimmerman DM., Brain Res., 1988, Vol. 454, p.68-72;Leander JD, Epilepsy Res., 1989 Vol. 4, p.28-33;and Yamaguchi S, Donevan SD, Rogawski MA., Epilepsy Res., 1993, Vol. 15, p.179-184]. If the animal did not show hind limb extension, it was considered to be protected from the convulsive effects of the electric shock. In some cases, the effective dose (ED) of the test or reference compound that abolished the tonic-extension component of the seizures in 50% of the animals was determined. 50And the 95% confidence interval) is calculated from the dose-response data [Litchfield, JT Jr, Wilcoxon., “A simplified method of evaluating dose-effect experiments.”, J. Pharmacol. Exp. Ther., 1949, Vol. 96, p.99-113]. For each experiment, one group of animals is always treated with vehicle (negative control), and one group of animals is always treated with the reference compound (positive control).

[0172] <AMPA receptor antagonist activity of the compound in vivo> The in vivo activity of AMPA receptor antagonist compounds can be assayed using mouse or rat maximal electroshock seizure (MES) model, or mouse or rat 6Hz psychomotor seizure model.The in vivo efficacy data of treatment with AMPA receptor antagonist tezampanel is shown in Ornstein PL et al., J Med. Chem, (USA), 1993, Vol. 36, p.2046-2048 in mouse MES model, and in Barton ME et al., Epilepsy Res, 2003, Vol. 56, p.17-26 in both mouse MES model and mouse 6Hz psychomotor seizure model. In vivo efficacy data for treatment of mice with the FDA-approved antiepileptic drug, AMPA receptor antagonist (AMPARA), perampanel, in both the mouse 6 Hz seizure model and the maximal electroshock seizure MES model is presented in Hanada T et al., Epilepsia, 2011, Vol. 52, p.1331-1340. In vivo efficacy data for the experimental AMPA receptor antagonist (AMPARA) compound YM928 tested in the MES mouse model is presented in Yamashita H et al., J Pharmacol Exp Ther, (US), 2004, Vol. 308, p.127-133. In vivo efficacy data comparing treatment of mice with the experimental therapeutic AMPARA compound YM928 and its derivatives in the MES model to the clinically studied AMPARA (talampanel) as a reference compound are presented in Inami H et al., Chem Pharm Bull (Tokyo), 2019, Vol. 67, p.699-706.

[0173] <References on the therapeutic efficacy of AMPA receptor antagonist compounds or drugs in the MES model or 6 Hz psychomotor seizure model in rats or mice> Ornstein PL, Arnold MB, Augenstein NK, Lodge D, Leander JD, Schoepp DD. 1993. J Med. Chem 36:2046-2048. competitive AMPA receptor antagonist" (Tezanpanel); Barton ME, Peters SC, Shannon HE. 2003. Epilepsy Res 56:17-26. "Comparison of the effect of glutamate receptor modulators in the 6 Hz and maximal electroshock seizure models" (Tezampanel); Hanada T, Hashizume Y, Tokuhara N, Takenaka O, Kohmura N, Ogasawara A, Hatakeyama S, Ohgoh M, Ueno M, Nishizawa Y. 2011. Epilepsia 52:1331-1340. "Perampanel: A Novel, Orally Active, Noncompetitive AMPA-receptor Antagonist That Reduces Seizure Activity in Rodent Models of Epilepsy" (Perampanel, an FDA-approved antiepileptic AMPA receptor antagonist, in mouse 6 Hz seizure models or MES models); Yamashita H, Ohno K, Amada Y, Hattori H, Ozawa-Funatsu Y, Toya T, Inami H, Shishikura JI, Sakamoto S, Okada M, Yamaguchi T. 2004. J Pharmacol Exp Ther 308:127-133. "Effects of 2-[N-(4-chlorophenyl)-N-methylamino]-4H-pyrido[3.2-e]-1,3-thiazin-4-one (YM928), an orally active alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor antagonist, in models of generalized epileptic seizure in mice and rats"(Experimental AMPA receptor antagonist (AMPARA) compound YM928 tested in the MES mouse model); Inami H, Shishikura JI, Yasunaga T, Hirano M, Kimura T, Yamashita H, Ohno K, Sakamoto S. 2019. Chem Pharm Bull (Tokyo) 67:699-706. “Synthesis and pharmacological evaluation of 3-[(4-oxo-4H-pyrido[3,2-e][1,3]thiazin-2-yl)(phenyl)amino]propanenitrile derivatives as orally active AMPA receptor antagonists”. (In vivo efficacy data comparing the experimental therapeutic AMPARA compound YM928 and its derivatives with reference to clinically studied AMPARA (talampanel) in the mouse MES model).

[0174] <Rotarod test for movement disorders> To verify whether the administered doses produced substantial motor impairment, the mice's motor performance on the rotarod was tested in parallel with the stimulation of the 6Hz psychomotor seizure model. Evaluation in the rotarod assay was performed immediately before the 6Hz psychomotor seizure model test. Thus, each treatment group was tested in the 6Hz psychomotor seizure model immediately after evaluation in the rotarod assay. When placed on a 1-inch knurled rod rotating at a speed of 6 rpm, the mice can maintain balance for an extended period of time. Motor impairment was assessed by determining whether the mice remained on the rotarod for a 1-minute observation period; i.e., 3 falls in 1 minute was considered as a failure on the rotarod.

[0175] Pharmacological observation of rat behavior for signs of motor impairment by trained observers. Rats were monitored after dosing for signs of impairment due to Compound 1 administered by the subcutaneous route at a dose of 6 mg / kg. No changes in tolerance or neurological changes were observed. All rats were alert, upright and behaving normally during the 30 minute observation period after dosing.

[0176] <Statistical analysis> Seizure protection and rotarod dysmotility data are presented as # (number of animals protected from seizures) / N (treatment group size consisting of the number of animals tested by treatment with vehicle or compound at a given dose) and # (number with dysmotility) / N (number tested), respectively. The dysmotility values ​​for a particular treatment group were compared to animals treated with VEH using Fisher's exact test. For dose-response analysis, the 50% efficacy ED 50 The doses resulting in values ​​(and 95% CI) were calculated using Prism (GraphPad Software) analysis, which used at least three treatment groups (N = 6–10 animals per group). Plasma concentrations are presented as mean ± SEM and were compared using Student's t test.

[0177] Table 3 shows the results of testing Compound 1 in an in vivo study of protection from seizures in a maximal electroshock convulsion (MES) model test in rats after a single acute systemic administration. Treatment with Compound 1 or vehicle was well tolerated by the rats.

[0178] Table 3: Results of treatment with Compound 1 in the maximal electroshock seizure (MES) model in rats. [Table 3]

[0179] Table 4 shows the results of testing compounds 1, 3, 4 and 6 in an in vivo study of protection from seizures in a 6 Hz psychomotor seizure model test in mice after a single acute systemic administration.

[0180] Table 4: Results of treatment with compounds 1, 3, 4, and 6 in a 6 Hz psychomotor seizure model in mice. [Table 4]

[0181] <Subcutaneous pentylenetetrazol (scPTZ) seizure model in rodents> The scPTZ model can be performed using mice or rats. The materials, methods and protocols are described in White HS, Johnson M, Wolf HH, Kupferberg HJ., “The early identification of anticonvulsant activity: role of the maximal electroshock and subcutaneous pentylenetetrazol seizure models”, Italian Journal Neurological Sciences, 1995, Vol. 16, p.73-77 (review) and in the following reviews (two chapters in two books): Levy RH, Mattson RH, Meldrum BS (eds.), “Antiepileptic Drugs”, (USA), Raven, New York, 4th ed., 1995, pp. 99-110 (White HS, Woodhead JH, Franklin MR. Swinyard EA, Wolf HH., “General principles: experimental selection, quantification, and evaluation of antiepileptic drugs.”); and Levy R, Mattson R, Meldrum B, Perucca E (eds.), “Antiepileptic Drugs”, (USA), Lippincott Williams and Wilkins, Philadelphia, 5th ed., 2002, pp. 36-48 (White HS, Woodhead JH, Wilcox KS, Stables JP, Kupferberg HJ, Wolf HH, “Discovery and preclinical development of antiepileptic drugs”).

[0182] <Lithium-pilocarpine-induced status epilepticus rat model> Status epilepticus can be induced in animals using either Long-Evans or Sprague-Dawley rats following published procedures (Metcalf CS, Radwanski PB, Bealer SL., “Status epilepticus produces chronic alterations in cardiac sympathovagal balance.”, Epilepsia. 2009, Vol. 50, No. 4, p.747-54;Clifford, DB, Olney, JW, Maniotis, A., Collins, RC, Zorumski, CF, “The functional anatomy and pathology of lithium-pilocarpine and high-dose pilocarpine seizures.”, Neuroscience, 1987, Vol. 23, p.953-968;Hanada T, Ido K, Kosasa T, “Effect of perampanel, a novel AMPA antagonist, on benzodiazepine-resistant status epilepticus in a (See Wu T, Ido K, Osada Y, Kotani S, Tamaoka A, Hanada T, “The neuroprotective effect of perampanel in lithium-pilocarpine rat seizure model,” Pharmacol. Res. Perspect. 2014, Vol. 2, No. 5, e00063; Wu T, Ido K, Osada Y, Kotani S, Tamaoka A, Hanada T, “The neuroprotective effect of perampanel in lithium-pilocarpine rat seizure model,” Epilepsy Res. 2017, Vol.137, p.152-158). The effects of the test compounds and their respective vehicles are compared to the reference compound perampanel or other reference compounds.

[0183] <Rodent pain models> The formalin pain test is based on a modification of the original model description (Malmberg AB, Yaksh TL, “Antinociceptive actions of spinal nonsteroidal anti-inflammatory agents on the formalin test in the rat”, J. Pharmacol Exp. Ther., 1992, Vol. 263, No. 1, p.136-46 and Wheeler-Aceto H, Porreca F, Cowan A, “The rat paw formalin test: comparison of noxious agents”, Pain, 1990, Vol. 40, No. 2, p.229-38). Briefly, minor modifications can be made to the above method: juvenile Sprague-Dawley rats are used (as described below) and formalin (Sigma) is prepared fresh daily in sterile saline.

[0184] Young adult male Sprague-Dawley rats (60–70 g body weight) were obtained from Charles River Laboratories (Wilmington, MA, USA). Animals had free access to food and water except during the study period. After delivery, animals were allowed sufficient time (~1 week) to acclimate to the housing conditions before the study and were housed in plastic cages in a room with controlled humidity, ventilation, and lighting (12 h on - 12 h off). Animals were housed in accordance with the recommendations of the Guide for Care and Use of Laboratory Animals (National Research Council) and in accordance with the guidelines established by the Institutional Animal Care and Use Committee (IACUC). Animal experiments were performed in accordance with the ARRIVE guidelines (UK). Protocols were pre-approved by the IACUC prior to the study.

[0185] Test compounds can be administered using a volume of fluid with an optimal ratio to body fluids. Test compounds can be administered in a volume of 0.04 mL per 10 g body weight in rats. Test compounds can be formulated into any of the formulations described above in the 6 Hz seizure model section. Morphine sulfate is commercially available from Sigma and can be prepared by compounding with saline. Morphine is typically administered via the subcutaneous (sc) route. Test substances or vehicles can be administered via the sc route of administration unless otherwise indicated. Other routes of administration (e.g. ip, sc, po, im, etc. can be used) and methods of compound administration can be used, including vehicle solutions and solvent formulations.

[0186] Formalin Pain Model in Rats. The formalin pain model can be performed as follows. Test compounds are administered to the animals 0.5 or 1 hour before a 5% formalin solution [prepared by mixing formalin (Sigma, St. Louis, MO, USA) in sterile saline] is injected subcutaneously (50 μL; 30-gauge needle) into the plantar region of the right hind paw of the rat. This formalin test is a well-established model for evaluating the analgesic effects of various compounds in mice and rats. Formalin induces a distinct biphasic behavioral profile characterized by an initial licking of the injected paw (Phase I: 0-10 min after formalin administration), followed by a brief period of reduced licking followed by a resumption of the licking behavior (Phase II: 20-45 min after formalin administration). After formalin injection, each animal is observed for the first 2 min of every other 5 min epoch until 45 min has elapsed. The cumulative length of licking during each 2 min period is measured. Area under the curve (AUC) values ​​for both Phase I and Phase II are normalized to the vehicle (VEH) treated group of rats.

[0187] An in vivo pharmacological method (patented by C. Pharmaceuticals, LLC) in male Sprague-Dawley rats (Charles River Laboratories) was used to study three different compounds in the study: Study I. In vivo acute dose PK in rats Study II. Determination of the in vivo SAD (single ascending dose study) maximum tolerated daily dose and bioanalytical determination of dose-exposure relationships in rats Study III. Determination of maximum tolerated dose and bioanalytical determination of dose-exposure relationships in a once daily 5-day MAD study in rats in vivo Used for.

[0188] Male Sprague-Dawley rats used in the above studies were supplied by Charles River Laboratories (Wilmington, MA, USA). Animals were housed in a veterinarian-certified animal care facility in the United States with a 7-day acclimation period. Upon arrival, rats weighed 225–250 g. At the time of the PK study, rats were 7–8 weeks old and weighed an average of 260–280 g. Rats were housed two per cage in a ventilated, HEPA-filtered cage-rack system. Rats were housed under a 12:12-h normal lighting cycle (12 h off, 12 h on, with lights on at 7:00 a.m. local time). For all studies, rats were provided with standard rodent chow and water ad libitum. Rats were not fasted during the study. Animals were handled prior to the study and randomly assigned to treatment groups by weight. The temperature in the housing and treatment rooms was 21-24°C, and the humidity was 40-42% (within the normal range for the facility) during the experiment. All procedures were completed in accordance with IACUC-approved protocols for housing and administering test substances.

[0189] <Administration of test substances (each compound formulated in an additive)> Intravenous (IV) Administration Route: To perform IV dosing, each rat was restrained in a decapicone® (a plastic bag with an open tip for breathing) with the tail clipped on either side to a table surface and left free to access the tail. A needle (23G x 1 inch, BD PrecisionGlide™, catalog #BD305145, manufacturer BD Biosciences, NJ, USA) attached to a 1 mL syringe (catalog #BD301025, manufacturer BD Biosciences, NJ, USA) was inserted into the lateral tail vein. Proper needle placement was ensured by withdrawing the syringe until just a trace of blood was visible. The required volume was then administered.

[0190] Oral Gavage (PO) Route To administer PO, each rat was restrained by immobilizing the head, neck, back, and upper body to prevent injury from struggling. Once in a stable vertical position, the rat was then dosed by inserting an oral probe (18G, Instech FTP-18-38, manufacturer Plymouth Meeting, PA, USA) attached to a 1 mL syringe (BD301025, manufacturer BD Biosciences, NJ, USA) or a 3 mL syringe (BD309588, manufacturer BD Biosciences, NJ, USA) into the rat's esophagus, depending on the required dose. Once properly positioned, the dose was expelled into the rat's stomach.

[0191] <Subcutaneous administration route> To perform SC dosing, each rat was restrained by fixing the head, neck, back, and upper body in a manner that allowed for "tenting" of the skin. Once in a stable position, a needle (21G x 0.5 inch, BD PrecisionGlide, BD305111, manufacturer: BD Biosciences, NJ, USA) attached to a 1 mL syringe (BD301025, manufacturer: BD Biosciences, NJ, USA) was inserted under the animal's fur into the tented area between the shoulder blades. The dose was then expelled under the rat's fur.

[0192] Blood was collected by tail bleed (0.0833, 0.25, 0.5, 1, 2, 4 hours for IV) and (0.5, 1, 1.167, 1.5, 2, 4, 6 hours for PO) and (0.25, 0.5, 1, 2, 4, 6 hours for SC). Terminal blood collection was performed by cardiac puncture 4 hours (IV) and 6 hours (PO and SC) post-dose. For rats undergoing Irwin's test (certain PO and SC groups as indicated), terminal blood collection was performed by cardiac puncture at 70 minutes. For rats in certain SC groups as indicated, terminal blood collection was performed by cardiac puncture at 30 minutes.

[0193] [Irwin Testing Procedure] Rats were subjected to a rat-adapted version of the Irwin test procedure, an in vivo assessment of the effects of test compounds on neurological function in rodents. Rats were allowed to habituate to the procedure room for at least 30 minutes. Rats were assessed for the following characteristics. Each rat was scored 0-3, with 0 representing a normal animal's response and 3 representing a maximally impaired animal. Please note that it is normal to score animals in the Irwin test in the range of 0-1. Animals were tested 55-60 minutes after administration of the test compound. References: Irwin, S., “Comprehensive observational assessment: la. A symptomatic quantitative procedure for assessing the behavioral and physiologic state of the mouse”, Psychopharmacologia, 20 September 1968, Vol. 13, No. 3, p.222-257.

[0194] <Irwin parameters - scoring> 1. Overall Appearance - Appearance Comments about grooming behavior, coat color, whiskers, etc. b. Scored from 0 to 3 i. 0: Normal - coat is smooth and shiny; eyes are wide open; no wounds. ii. 1: Slight dirt on the coat, slightly closed eyes iii. 2: Moderately dirty coat; eye discharge / closed eyes / skin lesions iv. 3: Dirty fur; bleeding on nails 2. Hyperactivity a. Cage observation b. Scored from 0 to 3, with 0 being absent and 3 being present i. 0: Normal activity; not hyperactive ii. 1: Touch or stimulation elicits a response greater than normal iii. 2: Grooming behavior is disrupted, running around the cage beyond normal exploratory behavior. iv. 3: Active running; jumping out of cage; no attempt to groom 3. Decreased activity a. Cage observation b. Scored from 0 to 3, with 0 being absent and 3 being present i. 0: Normal activity; not hypoactive ii. 1: Normal movement is slow and often stationary; stopped and no grooming behavior. iii. 2: Little to no movement in response to stimuli, remaining stationary rather than moving around iv. 3: No movement, no response to stimuli, stationary 4. Sedation a. Cage observation b. Scored from 0 to 3, with 0 being absent and 3 being present i. 0: Normal - no sedation ii. 1: Unsteady gait, staggering iii. 2: Partial eye closure iv. 3: Complete sedation; eyes closed 5. Seizures a. Scored from 0 to 3, with 0 being absent and 3 being present i. 0: no seizures ii. 1: Tremor (shivering) iii. 2:HIC - handling-induced convulsions iv. 3: Complete seizure - running-bouncing clonus or hind limb rigidity 6. Body position during rappelling a. The animal is placed in a tail-suspended position. The animals should exhibit struggling behavior and / or extension of all four limbs. b. Scored from 0 to 3, with no = 3 and yes = 0 i. 0: Struggling behavior and all four limbs extended ii. 1: Mild writhing behavior with some limbs extended iii. 2: Initially struggling, then immobility iv. 3: No struggling or limb movement 7. Crossed extensor reflex a. Pinching one of the hind legs causes the opposite hind leg to extend. b. Scored from 0 to 3, with no = 3 and yes = 0

[0195] 8. Forelimb / hindlimb placing response a. Touch the bottom of each paw in turn with a thin rod, and if the animal is suspended in the air and none of its other paws are touching a solid surface, it will lift its paw and place it back on the surface of the rod. b. Scored from 0 to 3, with no = 3 and yes = 0 9. Grasp reflex a. The animal grasps the tool when its paw is pecked b. Scored from 0 to 3, with no = 3 and yes = 0 i. 0: Grab the tool immediately ii. 3: No reaction when touching the feet 10. Bar Grabbing Behavior a. Place a rod for bar-grasping behavior on the animal's front paws to allow the animal to support its own weight. b. Scored from 0 to 3, with no = 3 and yes = 0 i. 0: Animals bear weight ii. 3: Animal does not attempt to hold on to the bars / does not stay on the bars after release 11. Righting reflex test a. Place the animal on its back and evaluate whether it turns over to lie on its stomach (normal position). b. Scored from 0 to 3, with no = 3 and yes = 0 i. 0: The baby flips over instantly and cannot be turned onto its back. ii. 1: The patient lies on their back but then quickly turns over iii. 2: Lying on one's back and slowly turning over or struggling to stand iv. 3: Not trying to recover 12. Tail-pinch reaction a. Animals respond to a tail pinch by withdrawing their body by pinching the tail ~1 cm from the base. b. Scored from 0 to 3, with no = 3 and yes = 0 i. 0: Reaction present, normal tail flick or escape reaction ii. 3: No reaction, no movement or escape 13. Auditory startle a. Reactions should be observed after the onset of auditory startle following the use of a dog clicker (a sound - emitting device). b. Score from 0 to 3 as: None = 3; Present = 0 (presence of startle, ear movement): i. 0: Present, normal startle or ear movement ii. 3: Absent, no startle reaction

[0196] <OFA. Measurement of open - field activity for spontaneous locomotor activity of living rats> Animals were allowed to habituate to the treatment room for at least 30 min prior to testing. All animals were dosed and placed in the open field chamber at the designated time post-dosing (immediately following the Irwin test) for a 15-min recording period (60-75 min after administration of the test substance treatment group (compound formulated in excipient or vehicle) or untreated control group). All open field experiments were performed in an "open field arena" (open field apparatus catalogue no. ENV-515 (manufacturer Med Associates Inc, Fairfax, VT, USA) which is a topless box made of clear organic glass with dimensions 43 × 43 × 30 cm (length × width × height, chamber internal dimensions). Each ENV-515 consists of 16 × 16 infrared photography beams with 2 cm spacing between beams to record movement in all three dimensions (x, y, z planes). Movements were recorded using the Med Associates designed Activity Monitor™ (Activity Monitor 7 software developed by Med Associates Inc, Fairfax, VT, USA) to measure endpoint stereotypy and locomotor counts. Distance traveled and rearing behavior were recorded (Activity Monitor 8 software developed by Med Associates Inc, Fairfax, VT, USA). (Monitor7 software). Reference: Open field activity in rodents and the effect of drug or test substance treatment: Fox KM, Sterling RC, Van Bockstaele EJ, “Cannabinoids and novelty investigation: influence of age and duration of exposure”, Behavioral Brain Research, (Netherlands), 2009, Vol. 196, p.248-253.

[0197] <Plasma and tissue collection> Blood was collected into K2EDTA tubes (BD Microtainer®, BD Biosciences, NJ, USA) and plasma was obtained by centrifuging the blood in a refrigerated centrifuge. Plasma was transferred to clean microcentrifuge tubes, frozen on dry ice, and stored at -80°C until bioanalysis. A minimum of 50 microliters of plasma was obtained. Whole brains were removed at the appropriate termination time points, weighed, and flash frozen on dry ice. Cerebrospinal fluid (CSF) was collected at the designated termination time points. CSF samples and brains were stored at -80°C.

[0198] <Blood collection procedure (for plasma)> Tail Bleeding Procedure: With rats in their home cages, the tail was carefully clipped (cut with scissors) approximately 1 cm from the tip. Blood was carefully collected into K2EDTA tubes using a BD microtainer. After retrieving the tubes, gentle pressure should be applied with cotton or gauze balls to stop bleeding.

[0199] Cardiac puncture - cardiac blood sampling procedure: Rats were anesthetized by placing them in a chamber containing CO2. The rats were subjected to the depth of anesthesia for euthanasia (sufficiently anesthetized). The animals were placed on their backs and the area above the sternum was rubbed with alcohol. The needle was placed under the skin just to the left of the sternum (left side for the animal). While holding the syringe at a 25-30 degree angle from the chest, a 10 mL needle (23G 1”, BD305145, manufacturer BD Biosciences, NJ, USA) was inserted 5 mm from the center of the chest towards the animal's lower jaw, 5-10 mm deep. The plunger was slightly withdrawn to create a slight negative pressure in the syringe. The needle was advanced to the midpoint of the thoracic cavity. When the needle reached the heart, blood flowed into the syringe and the sample was collected. The cardiac needle was kept in place and the plunger was gently withdrawn to collect the blood sample. The rat was euthanized by placing it back in the CO2 chamber.

[0200] <Cerebrospinal fluid (CSF) collection procedure> The animals were euthanized by placing them in a euthanasia chamber containing carbon dioxide. The rats were removed from the chamber and placed in sternal recumbency in the treatment area. The skin was cut with scissors from the upper part of the neck and spread along the midline to expose the cranial region of the skull. Using the fingers of one hand, the head was positioned so that it was bent downward at approximately 45°. CSF was collected by direct puncture into the cistern, using the occipital bone and the ala of the atlas as landmarks.

[0201] Suction on the syringe was released using a butterfly needle (EXELINT 21G x 3 / 4" Thin Wall, Catalog No. #277-04, Manufacturer: EXELINT, Redondo Beach, CA, USA). The needle was carefully supported (bevel up) with the opposite hand and gently "walked" or "slid" posteriorly toward the cerebral cistern (bony portion) carefully following the midline, over the occipital crest, until the surgeon could feel the needle enter the spinal cavity. Once correctly positioned, a small amount of negative pressure was applied to the syringe; resulting in CSF flowing into the needle hub (note that if CSF did not flow, the needle was repositioned and the procedure repeated). Negative pressure was applied with one hand until CSF flowed; the procedure was performed slowly to avoid blood contamination of the sample. Using a white paper background, the sample just above the needle was monitored for discoloration during collection (Note: once discoloration is observed, discontinue pressure on the syringe or tighten the butterfly needle just above the discoloration). To complete the CSF sample collection: the needle was removed from the syringe. The CSF sample was transferred from the syringe to a microcentrifuge tube (Note: once the blood was transferred to the centrifuge tube, it was centrifuged at 13,000 RPM in a clinical microcentrifuge for 1 minute). The CSF supernatant layer was collected, avoiding the blood at the bottom of the tube. The CSF was then quick frozen on dry ice and stored at -80°C until bioanalysis.

[0202] <Brain collection procedure> After CSF collection, the skull was rapidly cut open using scissors, and the brain was gently lifted using forceps and immediately flash frozen on dry ice and stored at −80°C until bioanalysis.

[0203] <Medication sample (DS) collection procedure> Compound 1 or compound 4 Dosing solutions or suspensions in excipients were collected after administration to the animals on each day. Dosing samples were flash frozen at -80°C until bioanalysis.

[0204] Compound 1 or compound 4 Quantitative LCMS bioanalysis of > Compound 1 or Compound 2 from rat samples (plasma, cerebrospinal fluid "CSF" or brain homogenate) 4 The measurements were performed at high resolution using a sensitive LCMS method.

[0205] Extracts of biological samples (either organic solvent precipitation or organic solvent extraction, Compound 1 or Compound 4 (i) Plasma or (ii) CSF or (iii) whole brain homogenate from rats treated with 1,2-dichlorophenyl ether (CD10) were run on LCMS and compared to freshly prepared standard curves on an identical LCMS instrument. The standard curves were generated by spike-in of known concentrations of Compound 1 or Compound 2 into rat plasma (supplied by Bioivt (formerly Bioreclamation), catalog number RAT00PLK2Y2N, Bioivt, Hicksville, NY, USA), such as male Sprague-Dawley rat plasma, collected in potassium-EDTA (K2-EDTA) tubes, centrifuged and stored at -80°C until use. 4 Standard LCMS (liquid chromatography-mass spectrometry) was performed using an AB Sciex Exion instrument and mass spectrometry (MS) detection on an API 5500 instrument. An autosampler AD multiplate was used (AB Sciex). The HPLC method used a Restek Force® Biphenyl column (dimensions 2.1×30 mm, particle size 1.8 micrometers) eluted with a gradient of two mobile phases (mobile phase A: 0.1% formic acid in water, and mobile phase B: acetonitrile with 0.1% formic acid) at a flow rate of 0.6 mL / min. Compound 1 or compound 2 were analyzed by HPLC. 4Tolbutamide was also used as an internal standard for MS detection and calibration. Compound 1 or compound 2 were detected based on the conditions used. 4 The lower limit of quantification was 1 ng / ml and the standard curve was linear up to 5,000 ng / ml. 4 The concentrations of compound were below the detection limit in vivo. 4 The concentration of compound was stable upon in vitro incubation in physiological buffer with or without fatty acid-free bovine serum albumin (Sigma-Aldrich, St. Louis, MO, USA) at 37°C for a 30 min incubation period. 4 In an in vitro study, compound 1 was hydrolyzed by rat plasma within a few minutes of incubation at 37°C. 4 Hydrolysis of was prevented by pretreatment of rat plasma with 10 μM of the serine hydrolase-esterase inhibitor methyl arachidonyl fluorophosphonate (supplied by Sigma-Aldrich) at 37°C for 5 min.

[0206] In vivo PK study of compounds 1 and 2 in male Sprague-Dawley rats Test Compound 1 or Test Compound 2 by the indicated route (either PO, SC or IV) 4 Rats were acutely dosed with 100 mg of ... 4 Each represents different rat treatment groups (A, B, C, D, E) administered various formulations (as specified) by various routes. Rats in treatment group A received SC administration of Cp#1 (formulated in saline with pH increased to pH 9.5 with NaOH; pH adjusted to 7.3 with HCl). Rats in treatment group B received SC administration of Cp#1 (formulated in saline with pH increased to pH 9.5 with NaOH; pH adjusted to 7.3 with HCl) by the intravenous (IV) route. Rats in treatment group C received SC administration of Cp#1 (formulated in saline with pH increased to pH 9.5 with NaOH; pH adjusted to 7.3 with HCl) by the PO route. compound 4HCl (prepared as a "solution" at a concentration of 10 mg / ml in 0.5% aqueous methylcellulose, 3% DMSO). Treatment group D rats were administered compound 4 HCl (prepared in 0.5% methylcellulose in water, 3% DMSO at a concentration of 10 mg / ml) was administered PO at a dose of 10 mg / kg and SC at a dose of 6 mg / kg. compound 4 It was administered from a solution.

[0207] Compounds in male Sprague-Dawley rats 4 In vivo SAD studies Single Ascending Dose Study (SAD) Test Compound by Oral PO Route 4 Rats were administered acute doses of 10 mg / kg of compound A for 1 day and samples were taken at the indicated time points for subsequent extraction, dissolution and quantitative LCMS bioanalysis. See Figure 7. For rat treatment groups A-D, the doses (A 10 mg / kg, B 20 mg / kg, C 30 mg / kg, D 50 mg / kg) were 4 The concentration of was 10 mg / ml, and the solution was prepared in 0.5% methylcellulose aqueous solution + 3% DMSO. compound 4 In the rat treatment groups E and F, the doses (E 100 mg / kg and F 200 mg / kg) were administered from a "solution" of 50 mg / ml of 0.5% methylcellulose in water + 3% DMSO. compound 4 The dose was administered from a "stable suspension" of 100 mg / kg.

[0208] The Irwin test in live rats was performed on the first (only test day) of the SAD study. 4 The data were collected 55-60 minutes after treatment with 10 mg / kg, 20 mg / kg, 30 mg / kg, 50 mg / kg, 100 mg / kg, and 200 mg / kg. Note: For endpoints, data are shown below each endpoint, shown on the x-axis, for increasing doses from left to right: 10 mg / kg, 20 mg / kg, 30 mg / kg, 50 mg / kg, 100 mg / kg, and 200 mg / kg. Many doses were normal or had very little change from normal, so no bar graphs are shown for the zero values.

[0209] OFA (open field activity) of live rats was measured using the test compounds in this 1-day test. 4 The results were obtained 60-75 minutes after treatment with . See Figures 8 and 9.

[0210] Compounds in male Sprague-Dawley rats 4 In vivo MAD test Repeated Ascending Dose Study (MAD) Test compounds were administered to rats once daily for 5 days and samples were taken at the indicated time points for subsequent extraction, dissolution and quantitative LCMS bioanalysis. For rat treatment groups A-D, doses (A 10 mg / kg, B 20 mg / kg, C 30 mg / kg, D 50 mg / kg) were administered at 10 mg / kg for compound A, 20 mg / kg for B, 30 mg / kg for C, and 50 mg / kg for D. 4 The concentration of was 10 mg / ml, and the solution was prepared in 0.5% methylcellulose aqueous solution + 3% DMSO. compound 4 In the rat treatment group with E, the dose (200 mg / kg E) was administered from a 50 mg / ml solution of 0.5% methylcellulose in water + 3% DMSO. compound 4 See Figures 15, 16, and 17. The Irwin test in vivo rats showed that the MAD study showed that the test compound was administered from a "stable suspension" of 100 mg / kg / day on two different treatment days (days 3 and 5). 4 The data were collected 55-60 minutes after treatment with 10 mg / kg, 20 mg / kg, 30 mg / kg, 50 mg / kg, 100 mg / kg, and 200 mg / kg. See Figures 13 and 14. Note: For endpoints, data are shown below each endpoint, shown on the x-axis, for increasing doses from left to right: 10 mg / kg, 20 mg / kg, 30 mg / kg, 50 mg / kg, 100 mg / kg, and 200 mg / kg. Many doses were normal or had very little change from normal, so no bar graphs are shown for the zero values.

[0211] OFA (open field activity) of adult rats was measured using the test compounds on two different treatment days (days 3 and 5). 4 The results were obtained 60-75 minutes after treatment with .

Claims

1. Formula I: 【Chemistry 1】 (wherein R is H and (C １ -C ２０ ) hydrocarbyl) A compound represented by the formula:

2. Formula II: 【Chemistry 2】 The compound according to claim 1 ,

3. R is H, and aliphatic, optionally substituted with 1 or 2 phenyl groups, provided that R contains 20 or fewer carbons (C １ -C ２０ 3. The compound of claim 2, wherein said alkyl group is selected from the group consisting of aryl, aryloxy ...

4. R is H or C ｎ H ｍ and wherein n is 1 and m is 3; n is 2 and m is 5; n is 3 and m is 3, 5, or 7; n is 4 and m is 5, 7, or 9; n is 5 and m is 7, 9, or 11; n is 6 and m is 5, 7, 9, 11, or 13; n is 7 and m is 7, 9, 11, 13, or 15; n is 8 and m is 5, 7, 9, 11, 13, 15, or 17; n is 9 and m is 7, 9, 11, 13, 15, 17, or 19; n is 10 and m is 7, 9, 11, 13, 15, 17, 19, or 21; n is 11 and m is 9, 11, 13, 15, 17, 19, 21, or 23; n is 12 and m is 7, 9, 11, 13, 15, 17, 19, 21, 23, or 25; n is 13 and m is 9, 11, 13, 15, 17, 19, 21, 23, 25, or 27; n is 14 and m is 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, or 29; n is 15 and m is 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, or 31; n is 16 and m is 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, or 33; n is 17 and m is 11, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, or 35; n is 18 and m is 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, or 37; n is 19 and m is 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, or 39; or n is 20 and m is 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41; The compound according to claim 2.

5. R is H, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, 1-methylpropyl, 1-methyl-2-ethylbutyl, 2-ethylbutyl, 2-methylpropyl, tert-butyl, 2-methylcyclopropyl, 1-methylcyclopropyl, cyclobutyl, cyclopropylmethyl (i.e., 【Chemistry 3】 ), n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, cyclobutylmethyl (i.e., 【Chemistry 4】 ), 2-(cyclopropyl)ethyl (i.e. 【Chemistry 5】 ), cyclopentyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1,1,2-trimethylpropyl, 1,2,2-trimethylpropyl, 3-(cyclopropyl)propyl (i.e., 【Chemistry 6】 ), 2-(cyclobutyl)ethyl (i.e. 【Chemistry 7】 ), cyclopentylmethyl (i.e. 【Chemistry 8】 ), cyclohexyl, cyclohexylmethyl, 2-cyclohexylethyl, dicyclohexylmethyl, n-octyl, benzyl, diphenylmethyl, decyl, dodecyl, tetradecyl, hexadecyl, hexadecyl, octadecyl, octadec-9-enyl, octadeca-9,12-dienyl, 2-propylpentyl, 2-butylhexyl, 2-pentylheptyl, 2-hexyloctyl.

6. 6. The compound of claim 5, wherein R is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, 2-methylbutyl, and 2-ethylbutyl. 【Request 7】 【Chemical 9】 【Chemistry 10】 【Chemistry 11】 3. The compound of claim 2 having a structure selected from:

8. 【Chemical 12】 8. The compound of claim 7 having a structure selected from:

9. 【Chemical 13】 9. The compound of claim 8 having a structure selected from:

10.

14. 9. The compound of claim 8 having a structure selected from:

11.

15. 9. The compound of claim 8 having a structure selected from:

12.

16. 9. The compound of claim 8 having a structure selected from:

13. 13. A pharmaceutical composition comprising a pharma- ceutically acceptable carrier and a compound of claim 1 or a pharma- ceutically acceptable salt thereof.

14. 13. A pharmaceutical composition for the treatment of a seizure disorder comprising a therapeutically or prophylactically effective amount of a compound according to claim 1.

15. The pharmaceutical composition of claim 14, wherein the seizure disorder is epilepsy.

16. The pharmaceutical composition of claim 14, wherein the seizure disorder is status epilepticus.

17. The pharmaceutical composition of claim 14, wherein the seizure disorder comprises partial onset seizures or primary generalized tonic-clonic seizures, or any combination thereof.

18. The pharmaceutical composition described in claim 14, wherein the seizure disorder is caused by a seizure disorder involving a hereditary gene.

19. The pharmaceutical composition of claim 14, wherein the seizure disorder is caused by a brain tumor, a concussive brain injury, or a penetrating brain injury.

20. 13. A pharmaceutical composition for the treatment of pain comprising a therapeutically or prophylactically effective amount of a compound according to claim 1.

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