Opioid compounds

Abstract

Compounds are represented by one of Formula I, wherein B may be of Formula B'. The compounds of Formula I may act as opioid reversal agents.

Description

OPIOID COMPOUNDSGOVERNMENT INTEREST

[0001] This presently-disclosed subject matter was made with government support under grant number DA051377 awarded by the National Institutes of Health. The government has certain rights in the presently disclosed subject matter.BACKGROUND

[0002] The presently-disclosed subject matter generally relates to opioid compounds, pharmaceutical compositions comprising such opioid compounds, methods of using such opioid compounds and compositions for treating or preventing pain in a subject, and methods of using such opioid compounds and compositions for harm-reduction goals in connection with opioid use disorder. In particular, certain embodiments of the presently-disclosed subject matter relate to structurally -unique opioid compounds having potent opioid agonist activity and reduced respiratory depression as compared to known opioids, such as fentanyl.

[0003] Although opioid antagonists, such as naloxone, have an established use in the treatment of opioid-induced respiratory depression, recent reports suggest that higher doses or repeated dosing of naloxone (due to recurrence of respiratory depression or renarcotization) may be required to fully reverse fentanyl-induced respiratory depression. Accordingly, there remains a need in the art for opioid reversal agents with greater efficacy as compared to known agents such as naloxone.BRIEF DESCRIPTION

[0004] Disclosed herein is a compound represented by one of Formula I,'* - - 'J\Rb)bFormula B'Formula Ior a co-crystal, hydrate, solvate, pharmaceutically acceptable salt or combination thereof, wherein:A and C are rings;B is Ci-8 alkyd optionally substituted with at least one Rb, or is represented by a ring of Formula B’;rings A, B’, and C are each independently i) a first ring, ii) a second ring, iii) a first ring fused to another first ring, iv) a second ring fused to another second ring, or (v) a first110712877:vlring fused to a second ring, wherein the first ring comprises a 5-membered carbocyclic ring, or a 5-membered heterocyclic ring, and the second ring comprises a 6-membered carbocyclic ring, or a 6-membered heterocyclic ring;a, b, and c are each independently 0 to 5, wherein the sum of a, b, and c is at least 1, Ra, b, and Rc are each independently Z, a Ce-Cso ether, halogen, deuterium, OH, CN, C1-C12 alkyl, C2-C12 alkenyl. C2-C12 alkynyl, C3-C6 cycloalkyl, Ci-C& alkyl(C3- Ct, cycloalkyl). Ci-Cs alkyllXX-Ce cycloalkenyl). C3-C6 cycloalkenyl. CX-Cr, heteroc cloalkyl, C -C6 heterocycloalkenyl, Ci-Cs alkyl(C2-C6 heterocycloalkyl), Ci-C's alkyl(C2-C6heterocycloalkenyl), C5-C12 aryl, C2-C12 heteroaryl, Ci-Cs alkyl (C2-C12 heteroaryl), each optionally substituted with deuterium, halogen, OH, CN, C1-C3 alkyl, C1-C3 haloalkyl, or a combination thereof, wherein any methylene may be replaced with and O or S;each occurrence of Rio is independently hydrogen, deuterium, Ci-Cg alkyl in which anrcarbon-carbon single bond is optionally replaced by a carbon-carbon double or triple bond, and the Ci-Cg alkyl is optionally substituted with deuterium, halogen, hydroxyl, cyano, or a combination thereof,provided that (i) the compound of Formula I comprises a C6-C30 ether; or (ii) the compound of Formula I comprises at least one Z; (iii) or the compound of Formula T comprises a C6-C30 ether and at least one Z;Z is Z’ or Yi-Z’, wherein: Z' is hydroxyl (-OH), thiol (-SH), cyano (-CN), fluorine (-F), isonitrile (-NC), nitro (-NO2). -X O(Y. Y NH2. -NH-Y2, -N(Y2)2, -O-Y2, -S-Y2, -S(=O)(Y2), -S(=O)(Y2)(NY2), -S(=O)2(Y2), aldehyde ( ( I h Ofi. -C(=O)Y2, -C(=S)Y2, -O-C(=O)Y2,-O-C(=S)Y2, -S-C(=O)Y2, -S-C(=S)Y2, -C(=O)-OY2, -C(=S)Y2, -C(=S)-OY2, - C(-O)-SY2, ••( •: S)-SY2. -O-C(= O:>-OY • -o-ct S)-OY2. -O-C Y O)-SY2, -O-( i SPSY-. -S- C(==O)-OY2, -S-C(:=:S)-OY2, -S-f{ O)-SY >. -S-C(=S)-SY2,-C(=O)-NH2, -( ( Oi-Xl lt Y2), - C(=O)-N(Y2)2. -C(=S)-NH2. -C(=S)-NH(Y2), -C(=S)-N(Y2)2, -NH-C(=O)-Y2, -N(Y2)-C(=O)-Y2, -NH-C(===S)-Y2, -N(Y.)-('( S;-Y. -NH-S(===O)2-Y2, -N(Y2)-S(=O)2-Y2, - O-C( =O)-NH 2, - O-C(==O)-NH(Y2), -O-C(==O)-N(Y2)2, -O-C(==S)-NH2, -O-C(-S -NH(Y2), -O-C( ==S)-N( Y 2)2, - NH-C(=O)-NH2, -NH-C(-O)-NH(Y2), -NH-C(:==O)-N(Y2)2, -X! l-C! S i-Xl i ■. -NH-C(-S)- NH(Y2), -O-C(=S)-N(Y2)2, -NH(Y2)-C(=O)-NH2. -NH(Y2)-C(=O)- H(Y2), -NH(Y2)-C(=O)~ N(Y2)2, -NH(Y2)-C(=:=S)-NH2, -NH(Y2)-C(:==:S)-NH(Y2), -O-C( S j-Y.; Y -i ■. carboxylic acid (Ci 0)0111. sulfonic acid (-SO3H ), -C( 0)XH-XH ■. -C( SiXH-XH.. -S(==0)NH~NH2. - C(=0)NH-NH2, -C(=S)NH-NH2, or -C(=NH-NH2)Y2; each occurrence of Yi and each occurrence of Y2independently comprise C1-C12 alky l, C2-Ci2alkenyl. C2-Ci2alkynyl, C3- Cb cycloalkyl, Ci-Cg aJkyl(C3-Cs cycloalkyl). Ci-Ce alkyl(C3-C-6 cycloalkenyl), C3- 210712877:vlCt, cycloalkenyl, C2-C6 heterocycloalkyl, C2-C6 heterocycloalkenyl. Cj-Ce alk l(C2- C« heterocycloalkyl), Ci-Ce alkyl(C2-Ce heterocycloalkenyl), C5-C12 aryl, C2-C12 heteroaryl, Ci-C« alkyl (C2-C12 heteroaryl), each optionally substituted with halogen, deuterium, or a combination thereof:Laand Lb are each a single bond or Ct-Ce alkyl in which any carbon-carbon single bond is optionally replaced by a carbon-carbon double or triple bond, any methylene is optionally replaced by O, S, Si, Ge, P, C(=O), S(=O), or NR10, and Laand Lb are each optionally substituted with Z, deuterium, halogen, OH, CN, C1-C3 alkyl, C1-C3 haloalkyl, or a combination thereof, provided that Laand Li, are not simultaneously a single bond.

[0005] Disclosed herein is a pharmaceutical composition, comprising an above¬ referenced compound of Formula I and a pharmaceutically acceptable excipient.

[0006] Disclosed herein is a method for treating an opioid overdose in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of the above-referenced compound.

[0007] Disclosed herein is a method for treating a subject having an opioid use disorder according to a harm reduction protocol, comprising administering a therapeutically effective amount of the above-referenced compound to the patient having an opioid use disorder.BRIEF DESCRIPTION OF THE DRAWINGS

[0008] The following figures are exemplary embodiments.

[0009] FIG. 1A is a graph illustrating the % inhibition of cAMP accumulation versus the logarithm (log) of fentanyl concentration in molarity (M) for a full dose response curve of fentanyl in the absence or presence of naltrexone (NTX) of Example 2;

[0010] FIG. IB is a graph illustrating the % inhibition of cAMP accumulation versus the log of fentanyl concentration (M) for a full dose response curve of fentanyl in the absence or presence of naloxone (NLX) of Example 2;

[0011] FIG. 1C is a graph illustrating the % inhibition of cAMP accumulation versus the log of fentanyl concentration (M) for a full dose response curve of fentanyl in the absence or presence of Compound 9 of Example 2;

[0012] FIG. ID is a graph illustrating the log of (A7A-1) versus the log of the concentration of the antagonist (M) for the Schild regression of the full dose response curve of fentanyl for NTX, NLX, and Compound 9 of Example 2;

[0013] FIG. IE is a table of the Schild analy sis for the full dose response curve of fentanyl for NTX, NLX, and Compound 9 of Example 2;310712877:vl

[0014] FIG. 2A is a graph illustrating the NLX (%) versus the log of the drug concentration (M) for the MOR-Gil -antagonist mode of Example 3;

[0015] FIG. 2B is a graph illustrating the NLX (%) versus the log of the drug concentration (M) for the MOR-Parrestin2-antagonist mode of Example 3;

[0016] FIG. 3A is a graph illustrating the experimental timeline for Example 4;

[0017] FIG. 3B is a graph illustrating the maximal possible of effect (MPE) ± standard error of the mean (SEM) (%) versus the log of the antagonist concentration (milligrams per kilogram, mg / kg) of Example 4;

[0018] FIG. 4A is a graph illustrating the experimental timeline for Example 5;

[0019] FIG. 4B is a graph illustrating the cumulative withdraw al score for the samples of Example 5;

[0020] FIG. 4C is a graph illustrating the cumulative withdrawal score for the samples of Example 5;

[0021] FIG. 5A is a graph illustrating the experimental timeline for Example 6;

[0022] FIG. 5B is a graph illustrating the distance traveled (centimeters, cm) versus the dose concentration of Compound 9 (mg / kg, subcutaneous) of Example 6:

[0023] FIG. 5C is a graph illustrating the time to right (seconds, sec) versus the dose concentration of Compound 9 (mg / kg, subcutaneous) of Example 6;

[0024] FIG. 6A is a graph illustrating the tidal volume (milliliters per breath per gram, mL / b / g) versus the dose concentration of Compound 9 (mg / kg, subcutaneous) of Example 6:

[0025] FIG. 6B is a graph illustrating the breaths per minute versus the dose concentration of Compound 9 (mg / kg, subcutaneous) of Example 6;

[0026] FIG. 6C is a graph illustrating the minute ventilation (milliliters per minute per gram, mL / min / g) versus the dose concentration of Compound 9 (mg / kg, subcutaneous) of Example 6;

[0027] FIG. 7A is a graph illustrating the experimental timeline for Example 6;

[0028] FIG. 7B is a graph illustrating the distance traveled (mean of cm ± SEM) versus the dose concentration of Compound 9 (mg / kg, subcutaneous) pre-plethysmography testing of Example 6;

[0029] FIG. 7C is a graph illustrating the distance traveled (mean of cm ± SEM) versus the dose concentration of Compound 9 (mg / kg, subcutaneous) post-plethysmography testing of Example 6;

[0030] FIG. 7D is a graph illustrating the time to right (sec) versus the dose concentration of Compound 9 (mg / kg, subcutaneous) pre-plethysmography testing of Example 6;410712877:vl

[0031] FIG. 7E is a graph illustrating the time to right (sec) versus the dose concentration of Compound 9 (mg / kg, subcutaneous) post-plethysmography testing of Example 6;

[0032] FIG. 8A is a graph illustrating the tidal volume (mL / b / g) versus the dose concentration of Compound 9 (mg / kg, subcutaneous) of Example 6;

[0033] FIG. 8B is a graph illustrating the breaths per minute versus the dose concentration of Compound 9 (mg / kg, subcutaneous) of Example 6;

[0034] FIG. 8C is a graph illustrating the minute ventilation (mL / min / g) versus dose concentration of Compound 9 (mg / kg, subcutaneous) of Example 6;

[0035] FIG. 9 is a graph illustrating the minute ventilation (mL / min / g) versus the rescue agent (mg / kg, intravenous) of Example 6; and

[0036] FIG. 10 is a graph illustrating the duration of vocal cord closure (sec) versus the intravenous (i.v.) rescue samples of Example 8.DETAILED DESCRIPTION

[0037] One of the most prominent opioid analgesics in the United States is the high potency agonist fentanyl. It is used in the treatment of acute and chronic pain and as an anesthetic adjuvant. Fentanyl was originally synthesized in 1960 and is approximately 100 times more potent than morphine with an LD50 of 3.1 mg / kg in rats, 0.03 mg / kg in monkeys, and unknown LD50 in humans. Among clinicians, fentanyl rapidly replaced morphine as an anesthetic for surgeries during the 1970s due to its more rapid onset, higher potency, and limited cardiovascular risks compared to morphine. Currently, there are several FDA-approved fentanyl analogues for medical and veterinary purposes, including the ultra-potent carfentanil (approximately 10,000 times more potent than morphine).

[0038] Chemically, synthetic opioids are highly toxic organic solids that may be encountered as injectable powders, liquids, nasal sprays, dermal patches and pills. The particle size of synthetic opioid powders typically ranges from 0.2 to 2.0 microns, and the powders are easily aerosolized, presenting a respiratory hazard and danger to water supply. A secondary dermal hazard exists if there is direct skin contact with large bulk amounts of concentrated threat materials. Due to its high potency, ingestion of just a few milligrams of fentanyl or other synthetic opioid can be deadly to an opioid-naive individual who is exposed to a aerosolized chemical attack or an unsuspecting '‘recreational” drug user upon acute exposure. Furthermore, first responders who come in contact with free base fentanyl analogues are at significant risk for life-threatening toxicities.

[0039] Currently, there are three opioid antagonists available on the market that have potential to reverse the effects of fentanyl in humans: naloxone (NLX), naltrexone (NTX),510712877:vland nalmefene (NLM). NLX is approved for administration by a variety of routes, including intravenous, intramuscular, subcutaneous and intranasal; sublingual and buccal formulations are under development. Under ordinary circumstances, NLX and NTX produce few effects in the absence of an exogenous agonist. However, there are certain conditions (e.g. shock) when the endogenous opioid systems are activated when administration of an opioid antagonist alone may have positive effects on hemodynamic changes. Administration of NLX or other opioid antagonist to a subject who is dependent on opioids can precipitate a withdrawal syndrome. In fact, it is this significantly aversive withdrawal state that can drive resumption of opioid use to mitigate withdrawal.

[0040] There have been reports that suggest that higher doses or repeated dosing of NLX (due to recurrence of respiratory depression) may be required to reverse fully fentanyl- induced respiratory depression. These findings have also been confirmed in mice where NLX less readily reversed respiratory depression by fentanyl compared with morphine. The lower sensitivity of fentanyl to antagonism by NLX cannot be explained simply by fentanyl having higher affinity for the MOR given that under competitive conditions, since the degree of antagonism does not depend on the affinity of the agonist but only upon the affinity and concentration of antagonist.

[0041] It is becoming increasingly clear that the currently approved synthetic opioid rescue agents have significant drawbacks and new agents are needed. One strategy for developing a new rescue agent against synthetic opioid exposure is to develop an opioid antagonist superior to NLX.

[0042] The presently-disclosed subject matter includes opioid compounds, pharmaceutical compositions comprising such opioid compounds, methods of using such opioid compounds and compositions for treating opioid overdose, and methods of using such opioid compounds and compositions for harm-reduction goals in connection with opioid use disorder. In some embodiments, the presently-disclosed subject matter includes structurally- unique opioid compounds having opioid antagonist activity.

[0043] The presently-disclosed compounds may be represented by a compound represented by Formula I,■' B'Formula B’Formula I610712877:vlor a co-crystal, hydrate, solvate, pharmaceutically acceptable salt, or combination thereof, wherein:A and C are rings;B is Ci-8 alkyl optionally substituted with at least one Rb, or is represented by a ring of Formula B’;rings A, B’, and C are each independently i) a first ring, ii) a second ring, iii) a first ring fused to another first ring, iv) a second ring fused to another second ring, or (v) a first ring fused to a second ring, wherein the first ring comprises a 5-membered carbocyclic ring, or a 5-membered heterocyclic ring, and the second ring comprises a 6-membered carbocyclic ring, or a 6-membered heterocyclic ring;a. b, and c are each independently 0 to 5, wherein the sum of a, b, and c is at least 1, Ra, Rb, and Rc are each independently Z, a C6-C30 ether, halogen, deuterium, OH, CN, C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl. C3-C6cycloalkyl, Ci-C6alkyl(C3- Ce cycloalkyl), Ci-Cs alkyl(C3-C« cycloalkenyl), C3-C6 cycloalkenyl, C2-C<5 heterocycloalkyl, C2-C6 heterocycloalkenyl, Ci-CY alkyl(C2-C6 heterocycloalkyl). Ci-Cr, alky 1(C2- C(, heterocycloalkenyl), C5-C12 aryl, C2-C12 heteroaryl, C1-C5 alkyl (C2-C12 heteroaryl), each optionally substituted with deuterium, halogen, OH, CN, C1-C3 alkyl, C1-C3 haloalky!, or a combination thereof, wherein any methylene may be replaced with O, S, Si, Ge, P, C(=O), St Or orNR’°;each occurrence of R10is independently hydrogen, deuterium. Ci-Cg alkyl in which any carbon-carbon single bond is optionally replaced by a carbon-carbon double or triple bond, and the Ci-Ce alkyl is optionally substituted with deuterium, halogen, hydroxyl, cyano, or a combination thereof,provided that (i) the compound of Formula I comprises a C6-C30 ether; or (ii) the compound of Formula I comprises at least one Z; (iii) or the compound of Formula I comprises a C6-C30 ether and at least one Z;Z is Z’ or Yi-Z’, wherein: Z’ is hydroxyl (-OH), thiol (-SH), cyano (-CN), fluorine (-F), isonitrile (-NC), nitro (-NO2). -N O( ¥.■). NH2. -NH-Y2, -N(Y2)2, -O-Y2, -S-Y2, -S(=O)(Y2). -S(=O)(Y2)(NY2), -S(=O)2(Y2), aldehyde (-CH(=O)). -C(=O)Y2. -C(=S)Y2, -O-Cl O? Y >.-O-C( SIY-. -S -( t () )Y2, -S-G S)Y.>. -G O)-GY>. -G S; Y?. -ft S;-OY >. - C(=O)-SY2, -C(==S)-SY2. -O-C(=O)-OY2, -O-C(=S)-OY2, -O-C(=O)-SY2, -O-C(=S)-SY2, -S- C(=O)-OY2, -S-C(=S)-OY2, -S-C(=O)-SY2, -S-C(=S)-SY2,-C(=O)-NH2, -C(=O)-NH(Y2), - C(=O)-N(Y2)2, -C(==S)-NH2, -C(===S)-NH(Y2), -C(===S)-N(Y2)2, -NH-C(==C))-Y2, -N(Y2)-C(=O)- Y2. -NH-C(=S)-Y2, -N(Y2)-C(=S)-Y2, -NH-S(=O)2-Y2, -N(Y2)-S(-O)2-Y2. -O-C(=O)-NH2, - 710712877:vlO-C(-O)-NH(Y2), -O-C(= O)-X(Y2) 2, -()-(■• 8 >-Yi! ■. -O-C(-S)~NH(Y2), -O-C(-S)~N(Y2)2„ - NH~C(-O)-NH2, -NH-C(==O)-NH(Y2), -NH-C(-O)-N(Y2)2, -NH-C(==S)-NH2, -NH-C(-S)~ NH(Y2), -()-( ( S)-X(Y2J:. -Nt K Y.>}-( ( =O)-Nt 12, -NH(Y2)-C(==O)-NH(Y2), -NH(Y2)-C(==O)- N(Y2)2. -NH(Y2)-C(-S)-NH2, -NH(Y2)-C(===S)-NH(Y2), -04?(= =S )-N( Y2)2, carboxylic acid (C(==O)OH), sulfonic acid (-SO3H), -C(-O)NH-NH2, -C(==S)NH-NH2, -S( OiYH-Xf I ■. -C(=O)NH-NH2, -C(=S)NH-NH2, or -C(=XH-NH2)Y2; each occurrence of Yt and each occurrence ofY2independently comprise C1-C12 alkyl. C2-C12 alkenyl, C2-C12 alkynyl, C3-Ce cycloalkyl, Ci-CX alkyllCfoCa cycloal yl), Ci-Ce alkyl(C?-C6 cycloalkenyl), C3- Co cycloalkenyl, Cz-Ce heterocycloalkyl, C2-Co heterocycloalkenyl, Ci-Ce alkyl(C2- Ce heterocycloalkyl). Cj-Ce alkyl(C2-C6 heterocycloalkenyl), C5-C12 aryl, C2-Ci2heteroaryl, Ci-Q> alkyl (C2-Ci2heteroaryl), each optionally substituted with halogen, deuterium, or a combination thereof;Laand Lb are each a single bond or Ct-Ce alky] in which any carbon-carbon single bond is optionally replaced by a carbon-carbon double or triple bond, any methylene is optionally replaced by O, S, Si, Ge, P, C(~O), S(-O), or NR10, and Laand Lb are each optionally substituted with Z, deuterium, halogen. OH, CN. C1-C3 alkyl, Cj-Cs haloalky I, or a combination thereof, provided that Laand Li, are not simultaneously a single bond.Definitions

[0044] As used herein, the term “alkyl” has the broadest meaning generally understood in the art, and may include a linear or branched moiety composed of carbon and hydrogen containing no double or triple bonds.

[0045] With respect to an optionally substituted moiety such as optionally substituted alkyl, a phrase such as “optionally substituted C1-12 alkyl” refers to a C1-12 alkyl that may be unsubstituted, or may have 1 or more substituents, and does not limit the number of carbon atoms in any substituent. Thus, for example, CH2(CH2)nOCH? is optionally substituted C1-12 alkyl because the parent alky 1 group has 12 carbon atoms. A phrase such as “C1-12 optionally substituted alkyl” refers to unsubstituted C1-12 alkyd, or substituted alky l wherein the alkyl parent and all substituents together have from 1-12 carbon atoms. For example, CH2CH2OCH3 is Ci-i2optionally substituted alkyl because the alkyl group (e.g. ethyl) and the substituent (e.g. methoxy) together contain 3 carbon atoms. Similar conventions may be applied to other optionally substituted moieties such as aryl and heterocyclyl.

[0046] As used herein the term “haloalkyl” has the broadest meaning understood in the art, and may include an alkyl group, as defined above, in which at least one hydrogen is replaced with a halogen selected from fluorine, chlorine, bromine and iodine. Examples of 810712877:vl“haloalkyl” may include fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, etc.

[0047] As used herein, the term "cycloalkyl” has the broadest meaning understood in the art, and may include a group having one or more saturated rings in which all ring members are carbon, such as cyclopropyl, cyclobutyl. cyclopentyl, cyclohexyl, etc.

[0048] As used herein, the term “halocycloalkyl” has the broadest meaning understood in the art, and may include a cycloalkyl group, as defined above, in which at least one hydrogen is replaced with a halogen selected from fluorine, chlorine, bromine and iodine. Examples of “halocycloalkyl” may include fluorocyclopropyl, difluorocyclopropyl, fluorocyclobutyl, difluorocyclobutyl, etc.

[0049] As used herein, the term “alkenyl” has the broadest meaning understood in the art, and may include a straight or branched chain, monovalent hydrocarbon group having at least one carbon-carbon double bond. Examples of “alkenyl” may include ethenyl, propenyl, butenyl, butadienyl, etc.

[0050] As used herein, the term “aryl” has the broadest meaning generally understood in the art, and may include an aromatic ring or aromatic ring system such as phenyl, naphthyl, dihydroindene, etc.

[0051] As used herein, the term “heterocycloalkyl” has the broadest meaning understood in the art, and may include a group having one or more saturated rings in which at least one ring member is a heteroatom. “Heterocycloalkenyl” means a cycloalkenyl group wherein one or more single bonds (e.g., carbon-carbon, carbon-heteroatom, or heteroatom-heteroatom) is replaced with a double bond. “Heterocycloalkynyl” means a cycloalkynyl group where one or more carbon-carbon single bonds is replaced with a carbon-carbon triple bond.

[0052] As used herein, the term “cycloalkyl” has the broadest meaning understood in the art, and may include a group having one or more saturated rings in which all ring members are carbon, such as, e.g. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.“Cycloalkenyl” means a cycloalkyl group wherein one or more carbon-carbon single bonds is replaced with a carbon-carbon double bond. Examples include cyclopropenyl, cyclobutenyl, and cyclopentenyl, cyclohexenyl. “Cycloalkynyl” means a cycloalkyl group where one or more carbon-carbon single bonds is replaced with a carbon-carbon triple bond. Examples include cyclopropynyl, cyclobutynyl, cyclopentynyl, and cyclohexynyl.

[0053] If stereochemistry is not indicated, a name or structural representation includes any stereoisomer or any mixture of stereoisomers and Applicant reserves the right to910712877:vlspecifically identify and claim a compound as a single stereoisomer or any particular mixture of stereoisomers.

[0054] Compounds described herein may contain an asymmetric center and may thus exist as enantiomers. Where the compounds according to embodiments herein possess two or more asymmetric centers, they may additionally exist as diastereomers. Embodiments herein include all such possible stereoisomers as substantially pure resolved enantiomers, racemic mixtures thereof, as well as mixtures of diastereomers. In some embodiments, the formulas are shown without a definitive stereochemistry at certain positions. Embodiments herein include all stereoisomers of such formulas and pharmaceutically acceptable salts thereof. Diastereoisomeric pairs of enantiomers may be separated by, for example, fractional crystallization from a suitable solvent, and the pair of enantiomers thus obtained may be separated into individual stereoisomers by conventional means, for example by the use of an optically active acid or base as a resolving agent or on a chiral HPLC column. Further, any enantiomer or diastereomer of a compound of the general formula may be obtained by stereospecific or stereoselective synthesis using optically pure or enantioenriched starting materials or reagents of known configuration. The scope of embodiments herein as described and claimed encompasses the racemic forms of the compounds as well as the individual enantiomers, diastereomers, and stereoisomer-enriched mixtures and Applicant reserves the right to specifically identify and claim a compound in any such form.

[0055] The compounds disclosed herein can exist as and therefore include all stereoisomers, conformational isomers and mixtures thereof in all proportions as well as isotopic forms such as deuterated compounds and Applicant reserves the right to specifically identify and claim a compound in any such form.

[0056] In some embodiments of the invention, one or more hydrogen atoms is replaced by a deuterium. It is well established that deuteration of physiologically active compounds offer the advantage of retaining the pharmacological profile of their hydrogen counterparts while positively impacting their metabolic outcome. Selective replacement of one or more hydrogen with deuterium, in a compound of the present invention, could improve the safety, tolerability and efficacy of the compound when compared to its all hydrogen counterpart.

[0057] Methods for incorporation of deuterium into compounds are well established. Using metabolic studies established in the art, the compound of the present invention can be tested to identify sites for selective placement of a deuterium isotope, wherein the isotope will not be metabolized. Moreover these studies identify sites of metabolism as the location where a deuterium atom would be placed.1010712877:vl

[0058] Ring A may be represented by one of Formulas A-1 to A-4, or one of Formulas A-1 to A-4 fused to another of Formulas A-1 to A-4, wherein the rings in the fused ring system may be the same or different; B may be represented by one of Formulas B-1 to B-4, or one of Formulas B-1 to B-4 fused to another of Formulas B-1 to B-4, wherein the rings in the fused ring system may be the same or different; and ring C may be represented by one of Formulas C-l to C-4, or one of Formulas C-l to C-4 fused to another of Formulas C-l to C- 4, wherein the rings in the fused ring system may be the same or different:A^A2,A3'A2 / A3-A2^2-63, 62-63 I A I HX A4A! A4 O / 1 BT IBiZB4Bl OyB4 A4. / A4~Zx4 / 1"4 A5A5A A As' AB B6-B5 B6-B5 A-1 A-2 A-3 A-4 B-1 B-3 B-4 crc2ZC3 C2^3-62 I C1C4 C, C40^1C4'CB c5-c6C5_C6C-1 C-3 C-4 wherein: Ai-Ae, Bi-Be, and Ci-Ce are each independently C, CH2, CH, CH(Ra), C(Ra), C(Ra)2. O, S, N, or NRi0, Bi-Be are each independently C, CH2. CH, CH(Rb). C(Rb), C(Rb)2, O, S, N. or NR10, and Ci-C6are each independently C, CH2, CH, CH(Rc), C(Rc), C(Rc)2, O, S, N, or NR10

[0059] In Formula I, rings A, B, and C may include: cyclopentane, cyclopentadiene, tetrahydrofuran, tetrahydro thiophene, cyclohexane, cyclohexadiene, tetrahydropyran, tetrahydrothiopyran, thioxane, dithiane, morpholine, thiomorpholine, sulfolane, furan, thiophene, pyrrole, silole, indene, benzofuran, benzothiophene, indole, benzosilole, oxazole, isoxazole, oxadiazole, isoxadiazole, oxatriazole, isoxatriazole, thiazole, isothiazole, thiadiazole, isothiadiazole, thiatriazole, isothiatriazole, pyrazole, imidazole, triazole, tetrazole, benzene, pyridine, pyrimidine, pyrazine, pyridazine, triazine, or fused combinations of the foregoing groups.

[0060] A single bond in the “A” ring (e.g., the A1-A2 bond, the A2-A3 bond, the A3-A4 bond, the A4-A5 bond, the As-Ae bond) may be replaced with a double bond, resulting in a cycloalkene ring as the " A” ring including at least one double bond. A single bond in the B ' ring (e.g., the B1-B2 bond, the B2-B3 bond, the B3-B4 bond, the B4-B5 bond, the B5-B6 bond) may be replaced with a double bond, resulting in a cycloalkene ring as the “B” ring that includes at least one double bond. A single bond in the “C” ring (e.g., the C1-C2 bond, the C2-C3 bond, the C3-C4 bond, the C4-C5 bond, the C5-C6 bond) may be replaced with a double bond, resulting in a cycloalkene ring as the ”C ' ring that includes at least one double bond.1110712877:vl

[0061] Referring to Formula I, at least one of ring A and ring B may be: iii) a first ring fused to another first ring, iv) a second ring fused to another second ring, or (v) a first ring fused to a second ring. In some aspects, ring A and ring B are each independently: iii) a first ring fused to another first ring, iv) a second ring fused to another second ring, or (v) a first ring fused to a second ring. In some aspects, at least one of ring A and ring B are each independently: (v) a first ring fused to a second ring. In some aspects, Laand Lb are each a single bond. Ring A may be represented by one of Formulas A-l to A-4, or one of Formulas A-l to A-4 fused to another of Formulas A-l to A-4, wherein the rings in the fused ring system may be the same or different; ring B may be represented by one of Formulas B-l to B-4, or one of Formulas B-l to B-4 fused to another of Formulas B-l to B-4, wherein the rings in the fused ring system may be the same or different; and ring C may be represented by one of Formulas C-l to C-4, wherein Ai-Ae are each independently C, CH2, CH, CH(Ra), C(Ra), C(R,)2, O, S. N, or NR10, BI-B6are each independently C, CH2, CH, CH(Rb), C(Rb), C(Rb)2, O, S, N, or NR10, and Ci-Ce are each independently C, CH2, CH, CH(Rc), C(Rc), C(RC)2, O. S, N, or NR!0.

[0062] In some aspects, compounds of Formula I include at least one Z. The sum of a, b, and c may be at least 2. at least one of Ra, Rb, and Rcis Z, and at least one of Ra, Rb, and Rcis a Ce-Cso ether. In some aspects, at least one of ring A and ring B are each independently a first ring fused to a second ring, at least one of Ra, Rb, and Rc is Z, and at least one of Ra, Rb, and Rccomprises a C6-C30 ether. In the foregoing aspects, the C6-C30 ether may be a polyether. In some aspects, the bond between the compound of Formula I and the C6-C30 ether may be either a carbon-oxygen bond or a carbon-carbon bond. In some aspects, when the C6-C30 ether is a poly ether, each polyether repeating unit may be “hydrocarbylene-O,” or “O-hydrocarbyl,” wherein “hydrocarbyl” and “hydrocarbylene” encompass groups including C and H only (e.g., alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and combinations thereof). In certain aspects, the poly ether is a C12-C18 poly ether and includes the repeating unit ((CH2)2O).

[0063] In some aspects, the Ce-Cso poly ether is attached to the compound of Formula I with a linking group other than a methylene or an oxygen of an ether unit of the poly ether. In some aspects, the Ce-Cso ether comprises the formula LG-polyether, wherein LG is a linking group. Exemplary linking groups include an ester, an amide, a urea, a carbonate, a carbamate, sulfur analogs of each, and the like. The linking groups can be in any orientation. For example, if the linking group is an ester, the ester may be attached to the compound of Formula I (e.g., attachment to A, B, B’, or C) via the oxygen of the ester or attachment to the 1210712877:vlcarbonyl carbon of the ester. In another aspect, there a hydrocarbyl group may be present in between the linking group (LG) and the compound of Formula I (e.g., attachment to A, B, B’, or C). Non-limiting examples of a hydrocarbyl group present between LG and the compound of Formula I include C1-3 alkyl, C2-4 alkenyl, and C2-4 alkynyl. In a preferred aspect when the linking group is present, the linking group may include an amide or an ester.

[0064] The compound of Formula I may be of one of Formulas I-A to I-GB2• B6I-DB'3. B?BSl-G wherein Ai-Ae are each independently C, CH, C(Ra), or N, Bi-Be are each independently C, CH, C(Rb), or N, and Ci-Ce are each independently C, CH, C(Rc), or N, and Xj-Xe are each independently CH2, CHRX, C(RX)2, N, O, S, C(=O), S(=O), S(=O)2, NH, or NR10.

[0065] The compound of Formula I may be represented by Formula IICI, C4, and C5 are each independently C, CH, C(Rc), or N, E is a Ce-Cio ether, and ring A, ring B'. Lb are as described for Formula I.

[0066] The “A ring” may be represented by one of the following formulas1310712877:vlwherein Ai, A2, and A4-A6 are each independently C, CH, C(Ra), or N, and X1-X3 are each independently CH2, CHRX, C(RX)2, N, O, S, C(=O), S(=O), S(=O)2, NH, or NR10. Rxmay be Z, a C6-C30 ether, halogen, deuterium, OH, CN, C1-C12 alkyl, C2- 2 alkenyl. C2-C12 alkynyl, C3-C6 cycloalkyl, C1-C.5 alkyI(C3-C6 cycloalkyl), Cj-Ce alkyl(C3-C6cycloalkenyl), C3- Cr, cycloalkenyl, C2-C6 heterocycloalkyl, C2-Cs heterocycloalkenyl, Ci-CV, alkyl (C2- Ce heterocycloalkyl), Ci-C<, alkyl(C2-C6 heterocycloalkenyl), C5-C12 and, C2-C12 heteroaryl, Ci-Cs alkyl (C2-Ci2heteroaryl), each optionally substituted with deuterium, halogen, OH, CN, C1-C3 allcy l. C1-C3 haloalkyl, or a combination thereof, wherein any methylene may be replaced with O, S, Si, Ge, P. C(=O), S(=O). or NR10.

[0067] The “B ring” may be represented by one of the following formulaswherein Bi-Be are each independently C, CH, C(Rb), or N, and X4-X6 are each independently CH2, CHRx, C(RX)2, N. O, S. C(=O). S(=O), S(=O)2, NH, or NR10.

[0068] In the foregoing formulas, Z is Z' or Yi-Z’, wherein Z' is -C(=O)-NH2, -C(=O)-NH(Y2), -C{ O:-X( Y -( ( SiAl b. -C(-S)-NH(Y2), -Ci M-XlY -S(-O)(Y2)(NY2), S(=:O)2(Y2)(NY2), NH-S(:=:O)2-Y2, or -N(Y2)-S(=O)2-Y2. Each occurrence of Yj and each occurrence of Y2may independently comprise Cj-Ce alkyl, C2-CG alkenyl, C2-Ce alkynyl, Cs-Cs cycloalkyl, Ci-Ce alkyRCs-Cg cycloalkyl), Ci-Cs alkyl(C3-C6 cycloalkenyl), or C3-C(, cycloalkenyl. In some aspects, Z is Z’ or Yi-Z’, wherein Z’ is Z’ is -C(::::())" NH2, -C(=O)- NH(Y2), -C( O)-\YY ) -( ■ Si-Yl I.-. -C(==S)-NH(Y2), -C(==S)-N(Y2)2, -S(==O)(Y2)(NY2), S(=O)2(Y2)(NY2), NH-S(=O)2-Y2, or -N(Y2)-S(=O)2-Y2, and each occurrence of Yi and each occurrence of Y2may independently comprise Cj-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C4-C6 cycloalkyl, C1-C3 alkyI(C -C6 cycloalkyl), Cj-Cs alkvliCX-Cg cycloalkenyl), or C4- 1410712877:vlCt, cycloalkenyl. In some aspects, Z is Z’ or Yi-Z', wherein Z’ is Z’ is -C(::::O)~NFl2, -C(=O)- NH(Y2), -C(==O)-N(Y2)2. -C(:=:S)~NH2. -C(=S)-NH(Y2), -€ =S )-N( Y2) -S(= =O)(:2)( N Y2), S(=O)2(Y2)(NY2), NH-StyO)2-Y2. or -N(Y2)-S(:==:O)2-Y2, and each occurrence of Yi and each occurrence of Y2may independently comprise Cj-( 4 alkyl.

[0069] In any of the foregoing formulas, each occurrence of Yi and each occurrence of Y2may independently comprise C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyi, C3-Ct, cycloalkyl, Ci-Ce alkyl(C3-Ce cycloalkyl), Ci-Ct, alkyltCs-Ct. cycloalkenyl), or C3-Ce cycloalkenyl. In some aspects, each occurrence ofYi and each occurrence of Y2may independently comprise Ci-Ce alkyl, C2-Ce alkenyl, C2-C.:, alkynyi, C3-C6 cycloalkyl, Cj-Ce alkyl(C?," C6 cycloalkyl), Ci-Ce alkyl(C3-Ce cycloalkenyl), or C3-C6 cycloalkenyl. In some aspects, each occurrence of Yi and each occurrence of Y2may independently comprise Ci-C4 alkyl. C2-C4 alkenyl, C2-C4 alkynyi, C4-C6 cycloalkyl, Ci-Cs alkyl(C4-Cg cycloalky]), Ci-C alkyl(C4-C6 cycloalkenyl), or C4-C6 cycloalkenyl. In some aspects, each occurrence ofYj and each occurrence of Y2may independently comprise C1-C4 alkyl.

[0070] In some aspects, the B group of the compound of Formula I may be a Ci-s alkyl group. In some aspects, the C1-8 alkyl is a C3-8 alkyl. In some aspects, the C3-8 alkyl is a branched alkyl group, such as, for example, isopropyl, sec-butyl, tert-butyl, isobutyl, isopentyl, sec-pentyl, tert-pentyl, neopentyl, and the like. In such aspects, Laand Lb may each independently be a Ci-Ce alkyl in which any methylene is optionally replaced by C(=O), O, S, or NR10. In some aspects, one of Laand Lb is a C1-3 alkyl in which any methylene is optionally replaced by C(=O), O, S, or NR10and the other of Laand Lb is a C3-6 alkyl in which any methylene is optionally replaced by C(=O), O, S, or NR10. In some aspects, one of Laand Lb is a Ci alky l, optionally replaced by C(=O), O, S, or NR10and the other of Laand Lb is a C4-6 alkyd in which any methylene is optionally replaced by C(=O), O. S, or NR10. In some aspects, one of Laand Lb is O and the other of Laand Lb is (CH2)mN(R10)(CH2)n, wherein m and n are each 0 to 5, and the sum of m + n is at least 1. In the foregoing aspects, Z may be Z’.

[0071] In some aspects, the B group is represented by Formula B’ and at least one of A and B’ is a second ring (e.g.. 6-membered carbocyclic ring or 6-membered heterocyclic ring). In some aspects, A and B’ are each independently a second ring. In some aspects. A’ is a 6-membered aromatic or a 6-membered heteroaromatic ring and B’ is a 6-membered carbocyclic ring. In the foregoing aspects, Z may be Z’.

[0072] In some aspects of Formula I, Laand Lb may each independently be a Ci-Ce alkyl in which any methylene is optionally replaced by C(=O), O, S, or NR10. In some aspects, one 1510712877:vlof Laand Lb is a C1-3 alky l in which any methylene is optionally replaced by C(=O), O, S, or NR10and the other of Laand Lb is a C3-6 alkyl in which any methylene is optionally replaced by C(=O), O, S, or NR10. In some aspects, one of Laand Lb is aCi alkyl, optionally replaced by C(=O), O, S, or NR10and the other of Laand Lb is a C4-6 alkyl in which any methylene is optionally replaced by C(=O), O, S, or NR10. In some aspects, one Laand Lb is (CH2)mN(R10)(CH2)n, wherein m and n are each 0 to 5, and the sum of m + n is at least 1. In some aspects, one of Laand Lb is O and the other of Laand Lb is (CH2)mN(R10)(CH2)n, wherein m and n are each 0 to 5, and the sum of m + n is at least 1. In the foregoing aspects, any methylene is optionally substituted with deuterium, fluoro, or a combination thereof. In the foregoing aspects, Z may be Z’.

[0073] The compound of Formula I may be represented by Formula III.A2XA6I I — I—A3x.<A5 >"(Rb)bA4Formula III Formula B’

[0074] In Formula IIL Ai-Ag are each independently C, CH, C(Ra), orN, and Ci-Ce are each independently C, CH, C(Rc), or N. The other variables are the same as disclosed for Formulas I and II.

[0075] In Formula III, Formula B‘ may be represented by one of the following formulasB’-l B’-4wherein Bi-Be are each independently C, CH, or C(Rb), and X4-Xr, are each independently CH2, CHRX, C(RX)2, N, O, S, C(=O), S(=O), S(=O)2, NH, or NR10.

[0076] The compound of Formula III may be represented by Formula III-l or III-2:^AKA2XA6 I I — I—A3 <A5A4Formula III-1 Formula III-21610712877:vlwherein the variables are the same as described for Formulas I-III, B"’ is represented by Formula B’7, and Z is Z'.

[0077] The compound of Formula III may be represented by one of Formula III-A to III- D:Formula 111-A Formula III-BFormula III-C Formula III-Dwherein A1-A5 are each independently C, CH, C(Ra), or N; B1-B4 and Be are each independently C, CH, C(Rb); or N. Ci, C3, C4, and Ce are each independently C, CH, C(Rc), or N: and X4-X6 are each independently CH2, CHRX, or C(RX)2; R10is hydrogen, deuterium or C1-3 alkyl optionally substituted with deuterium, halogen, or a combination thereof; n is 0-4; wherein any hydrogen of a methylene is optionally replaced with deuterium, or fluoro. In some aspects, Z is present and is Z’.

[0078] In the foregoing aspects, Ra-Rc and Rxmay each independently include: Z, a C6- C30polyether, halogen, deuterium, OH, CN, C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyL C3- C6cycloalkyl, Ci-C6alkyl(C3-C.:, cycloalkyl), Ci-C6alkyl(C3-C6cycloalkenyl), C3- Ce cycloalkenyl, C2-C6 heterocycloalkyl, C2-C6 heterocycloalkenyl, C1-C6alkyl (C2-C6heterocycloalkyl), C1-C6alkyl(C2-C6heterocycloalkenyl), C5-C12 aryl, C2-C12heteroaryl, C1-C6alkyl (C2-C12heteroaryl), each optionally substituted with deuterium, halogen, OH, CN, C1-C3 alkyl, C1-C3 haloalkyl, or a combination thereof, wherein any methylene may be replaced with and O or S.

[0079] The compounds of Formula I may be one of the compounds below'1710712877:vlo9: X = N, R = H, R' = CH2CH2Ph; 10: X = CH, R = H, R' = CH2CH2Ph; 11: X = N, R = Cl, R' = CH2CH2Ph: 12: X = N, R = OCH3, R' = CH2CH2Ph; 13: X - N, R - OCH2Ph, R’ - CH2CH2Ph; 14: X = N, R = OCH3, R' = cyclohexylethyl 15: X = N, R = OCH3, R' = CH2CH2CH(CH3)2: 16: X - N, R - OCH3, R' - 2-indanylJWP-231JWP-230JWP-2711810712877:vl

[0080] The presently-disclosed subject matter further includes a pharmaceutical composition comprising a compound as disclosed herein and a pharmaceutically acceptable excipient.

[0081] The presently-disclosed subject matter further includes a method of using the compound as disclosed herein as an opioid antagonist.

[0082] The presently-disclosed subject matter further includes a method of treating opioid overdose in a subject comprising administering to the subject a therapeutically effective amount of a compound as disclosed herein.

[0083] The presently-disclosed subject matter further includes a method of treating a subject having an opioid use disorder according to a harm reduction protocol, comprising administering to the subject a therapeutically effective amount of a compound as disclosed herein.

[0084] While the terms used herein are believed to be well understood by those of ordinary skill in the art, certain definitions are set forth to facilitate explanation of the presently-disclosed subject matter.

[0085] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the presently disclosed subject matter(s) belong.1910712877:vl

[0086] All patents, patent applications, published applications and publications, GenBank sequences, databases, websites and other published materials referred to throughout the entire disclosure herein, unless noted otherwise, are incorporated by reference in their entirety.

[0087] As used herein, the abbreviations for any protective groups, amino acids and other compounds, are, unless indicated otherwise, in accord with their common usage, recognized abbreviations, or the IUPAC-IUB Commission on Biochemical Nomenclature (see, Biochem. (1972) 11(9): 1726-1732).

[0088] Although any methods, devices, and materials similar or equivalent to those described herein can be used in the practice or testing of the presently-disclosed subject matter, representative methods, devices, and materials are described herein.

[0089] The present application can ‘“comprise” (open ended) or “consist essentially of’ the components of the presently disclosed subject matter as well as other ingredients or elements described herein. As used herein, “comprising” is open ended and means the elements recited, or their equivalent in structure or function, plus any other element or elements which are not recited. The terms “having” and “including” are also to be construed as open ended unless the context suggests otherwise.

[0090] T he terms "a" and "an" do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item.

[0091] Unless otherwise indicated, all numbers expressing quantities of ingredients, properties such as reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about”. Accordingly, unless indicated to the contrary, the numerical parameters set forth in this specification and claims are approximations that can vary depending upon the desired properties sought to be obtained by the presently-disclosed subject matter.

[0092] As used herein, the term “about,” when referring to a value or to an amount of mass, weight, time, volume, concentration or percentage is meant to encompass variations of in some embodiments ±20%, in some embodiments ±10%, in some embodiments ±5%, in some embodiments ±1%, in some embodiments ±0.5%, in some embodiments ±0.1%, in some embodiments ±0.01%. and in some embodiments ±0.001% from the specified amount, as such variations are appropriate to perform the disclosed method.

[0093] Recitation of ranges of values are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. The endpoints of all ranges are included within the range and 2010712877:vlindependently combinable. All methods described herein can be performed in a suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or language indicating an example (e.g., "such as"), is intended merely for illustration and does not pose a limitation on the scope of the presently disclosed subject matter unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the presently disclosed subject matter.

[0094] " Pharmaceutically acceptable salts" includes derivatives of a compound of Formula I, wherein the a compound of Formula I is modified by making acid or base addition salts thereof, and further refers to pharmaceutically acceptable solvates, including hydrates, and co-crystals of such compounds and such salts. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid addition salts of basic residues such as amines; alkali or organic addition salts of acidic residues; and the like, and combinations comprising one or more of the foregoing salts. The pharmaceutically acceptable salts include non-toxic salts and the quaternary ammonium salts of the active agent, a compound of Formula I. For example, non-toxic acid salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; other acceptable inorganic salts include metal salts such as sodium salt, potassium salt, cesium salt, and the like; and alkaline earth metal salts, such as calcium salt, magnesium salt, and the like, and combinations comprising one or more of the foregoing salts. Pharmaceutically acceptable organic salts includes salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxy maleic, phenylacetic, glutamic, benzoic, salicylic, mesylic, esylic, besylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, HOOC-(CH2)n-COOH where n is 0-4, and the like; organic amine salts such as triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N, N'-dibenzylethylenediamine salt, and the like; and amino acid salts such as arginate, asparginate, glutamate, and the like; and combinations comprising one or more of the foregoing salts; organic amine salts such as triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N, N' dibenzylethylenediamine salt, and the like; and amino acid salts such as arginate, asparginate, glutamate, and the like; and combinations comprising one or more of the foregoing salts.2110712877:vl

[0095] A “pharmaceutically acceptable excipient” means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable, and includes an excipient that is acceptable for veterinary use as well as human pharmaceutical use. A “pharmaceutically acceptable excipient” as used in the present application includes both one and more than one such excipient.

[0096] A “patient” is a human or non-human animal in need of medical treatment.Medical treatment can include treatment of an existing condition, such as a disease or disorder, prophylactic or preventative treatment, or diagnostic treatment. In some embodiments, the patient is a human patient.

[0097] A “therapeutically effective amount” of a pharmaceutical combination of this disclosure means an amount effective, when administered to a patient, to provide a therapeutic benefit such as an amelioration of symptoms, e.g., an amount effective to decrease the symptoms of a pain.

[0098] The presently-disclosed subject matter is further illustrated by the following specific but non-limiting examples. The following examples may include compilations of data that are representative of data gathered at various times during the course of development and experimentation related to the presently disclosed subject matter.

[0099] As used herein, “carbocyclic ring,” refers to a saturated or unsaturated cyclic group having carbon atoms only as ring-forming atoms. As used herein, “heterocyclic ring,” refers to a saturated or unsaturated cyclic group having carbon atoms and at least one heteroatom as ring forming atoms. The term “alkyl” refers to a linear or branched saturated aliphatic hydrocarbon group “Alkenyl” refers to an alkyl group wherein a carbon-carbon single bond is replaced with a carbon-carbon double bond. “ Alkynyl” refers to an alkyl group wherein a carbon-carbon single bond is replaced with a carbon-carbon triple bond. The term “cycloalkyl” refers to a saturated hydrocarbon monocyclic group. “Cycloalkenyl” refers to a cycloalkyl group wherein a carbon-carbon single bond (in the ring) is replaced with a carbon-carbon double bond. “Heterocycloalkyl” refers to a cycloalkyl group having carbon atoms and at least one heteroatom as ring forming atoms. “Heterocycloalkenyl” refers to a heterocycloalkyl group wherein a carbon-carbon single bond in the ring is replaced with a carbon-carbon double bond “Aryl” refers to a carbocyclic aromatic ring. “Heteroaryl” refers to an aromatic ring including carbon atoms and at least one heteroatom as ring forming atoms. The term “alkylQC)” refers to an alkylene group substituted with an “X” group. For2210712877:vlexample, “Cj-Ce aikyKCs-Ce cycloalkyl)” refers to a Ci-CT alkylene (divalent) group substituted with a Ca-Cs cycloalkyl group.

[0100] The disclosure also includes a compound of Formula L or any formula disclosed herein, in which from 1 to “n” hydrogens attached to a carbon atom is / are replaced by deuterium, in which n is the number of hydrogens in the molecule. Such compounds exhibit increased resistance to metabolism and are thus useful for increasing the half-life of any compound of Formula I when administered to a subject. Such compounds are synthesized by means well known in the art. for example by employing starting materials in which one or more hydrogen atoms have been replaced by deuterium

[0101] The term “hydrate” refers to the complex formed by the combining of a compound of Formula I, or any formula disclosed herein, and water. The term “solvate” refers to a complex formed by the combining of a compound of Formula L or any formula disclosed herein, and a solvent or a cr s tai line solid containing amounts of a solvent incorporated within the crystal structure. The term “co-crystal” refers to a cry stalline material formed by combining compound of Formula I, or any formula disclosed herein, and one or more co-crystal formers (i.e., a molecule, ion or atom). In certain instances, co-crystals may have improved properties as compared to the parent form (i.e., the free molecule, zwitterion, etc.) or a salt of the parent compound. Improved properties can be increased solubility, increased dissolution, increased bioavailability, increased dose response, decreased hygroscopicity, a crystalline form of a normally amorphous compound, a crystalline form of a difficult to salt or unsalable compound, decreased form diversity, more desired morphology, and the like. Methods for making and characterizing co-crystals are known to those of skill in the art.

[0102] This disclosure is further illustrated by the following examples, which are non-limiting.EXAMPLESGeneral Experimental Procedures.

[0103] Reactions were performed in oven-dried glassware under normal atmosphere (unless otherwise specified). All chemical reagents w ere purchased from commercial suppliers and used without further purification. Anhydrous solvents w ere either purchased from commercial suppliers or obtained from a solvent purification system in which solvent w as passed through two columns of activated alumina under nitrogen. Reactions were monitored by thin-layer chromatography (TLC) on 0.25 mm Analtech GHLF silica gel plates 2310712877:vland visualized by UV (254 nm). Flash column chromatography was performed on a CombiFlash NextGen 300+ purification system (Teledyne Isco) using prepacked RediSepRf silica columns. Melting points were determined on a Thermo 100 ID Mel-Temp Electrothermal Melting Point Apparatus. 'H and13C NMR were recorded on a Bruker 400 megahertz (MHz) spectrometer, 500 MHz Varian spectrometer, and / or Bruker 600 MHz spectrometer. Chemical shifts are reported in parts per million (ppm) and referenced with respect to residual solvent: CDCh at 7.26 ppm, MeOD at 3.31 ppm, DMSO-d6 at 2.50 ppm (¹H NMR); CDCl3 at 77.16 ppm, MeOD at 49.00 ppm, DMSO-d6 at 39.52 ppm (13C NMR). In other cases, CDCh containing tetramethylsilane (TMS) was utilized. Coupling constants (J) are reported in hertz (Hz). High-resolution mass spectra (HRMS) were obtained on an Agilent 6230 time-of-flight mass spectrometer with an electrospray ion source in positive mode. Compounds selected for biological testing were identified as > 95% pure by high performance liquid chromatography (HPLC) on an Agilent 1260 Infinity II with diode array detection at 214 nm. A Poroshell 120 EC-C18 column (4.6 x 100 mm, 2.7 pm) with a 5-minute gradient mobile phase of 10 - 100% acetonitrile / 0.1% TFA in water (method A) and an Eclipse XDB-C18 column (4.6 x 150 mm, 5 pm) with a 5-minute gradient mobile phase of 10 - 100% acetonitrile / 0.1 % H3PO4 in water (method B) were utilized.EXAMPLE 1. Aminobenzyloxyarylamides Preparation.CHOf-fcW ’’ a tT^X x ft ■>«».;<» N « » M •m>. X »CH th. X - SB.8 - « S. X * N. ft « ft, ft' »<». X 1M. tVX*& R C ft t «; W^ mX« $ R-“ OCHs U. X*ft,ft <x _> ft - tx x*x,ft v < # i«i3¥a«^.?*, X « ft. ft ( I * 1§. X » ft. ft < v, ft •• s^c^*c^x£ft*wU'-' ft • Reagents and conditions: (a) Appropriate 4-hydroxybenzaldehyde, K2CO3, DMF, 100 - 130 °C, 2 h; (b) K2CO3, DMSO, H2O2, 0 or 10 °C to room temperature, 2 h; (c) NaBH(OAc)3, AcOH, DCE, room temperature, 24 h.

[0104] Reaction of 6-fluoro-3-pyridinecarbonitrile (Compound 17a), 4-hydroxybenzaldehyde, and K2CO3 in dimethylformamide (DMF) provided 6-(4-formyl-phenoxy)-3-pyridinecarbonitrile (Compound 18a). Conversion to 6-(4-formylphenoxy)-3-pyridinecarboxamide (Compound 19a) was carried out with basic hydrogen peroxide.Reductive amination with 2-phenethylamine utilizing sodium triacetoxyborohydride and acetic acid in 1,2-dichloroethylene (DCE) provided Compound 9.

[0105] Compounds 10 - 16, were synthesized analogously to Compound 9.Synthesis of 6-(4-((Phenethylamino)methyl)phenoxy)nicotinamide (Compound 9).2410712877:vl

[0106] A mixture of powdered molecular sieves (3 A), 2-phenylethan-l -amine (2.22 grams (g), 18.33 millimoles (mmol)), Compound 19a (3.70 g. 15.27 mmol), and 1,2-dichloroethane (30 milliliters (mL)) was stirred at room temperature overnight. Sodium triacetoxyborohydride (8.09 g, 38.19 mmol) and acetic acid (2.2 mL, 38.19 mmol) were added and the reaction mixture was stirred for an additional 3 hours (h). The reaction was filtered through a pad of Celite, quenched with saturated aqueous NaHCO3 (75 mL), and then extracted with EtOAc (3 x 50 mL). The combined organic extracts were dried (Na2SO4). filtered, and then concentrated in vacuo. The resulting crude material was purified by flash chromatography eluting with 0 - 15% MeOH / EtOAc to afford 1.20 g (22.6%) of Compound 9 as off-white solid, melting point (mp): 147-148 °C. ¹H NMR (400 MHz, CDCl3) δ8.58 (d, J= 2.5 Hz, 1H), 8.16 (dd,.7= 8.6, 2.5 Hz, 1H), 7.38 - 7.27 (m, 4H), 7.25 - 7.17 (m, 3H), 7.13 - 7.05 (m, 2H), 6.96 (d, J= 8.6 Hz, 1H), 5.78 (br s, 2H), 3.82 (s, 2H), 2.94 (t, J= 7.2 Hz, 2H), 2.85 (t, J= 7.0 Hz, 2H). 13C NMR (101 MHz, CDCl3) δ 167.10, 166.16, 152.34, 147.33, 140.12, 139.43, 137.62, 129.59, 128.87, 128.62, 126.32, 124.19, 121.45, 111.27, 53.43, 50.76, 36.48. HRMS (ESI) m / z for [M + H]+calcd for C21H21N3O2, 348.1707; found, 348.1709. HPLC: retention time (tR) = 2.606 min; purity > 99.0%.Synthesis of 4-(4-((Phenethylamino)methyl)phenoxy)benzamide (Compound 10).

[0107] A mixture of powdered molecular sieves (3 A, 0.9 g), 2-phenylethan-l -amine (0.17 g, 1.39 mmol), Compound 19b (0.28 g, 1.16 mmol), and 1,2-di chloroethane (15 mL) was stirred at room temperature for 12 h. Sodium triacetoxyborohydride (0.61 g, 2.90 mmol) and acetic acid (0.17 g, 2.90 mmol) were added and the reaction was stirred for 12 h. The reaction was filtered through a pad of Celite, quenched with saturated aqueous NaHCO3 (30 mL), and then extracted with EtOAc (3 x 30 mL). The combined organic extracts were dried (Na2SO4), filtered, and then concentrated in vacuo. The resulting crude material was purified by flash chromatography eluting with 0 - 8% MeOH / EtOAc to afford 0.05 g (13%) of 10 as peach colored solid, mp: 104-105 °C. ¹H NMR (400 MHz, CDCl3) δ 7.81 - 7.74 (m, 2H), 7.30 (m, 4H), 7.21 (m, 3H), 7.03 - 6.95 (m, 4H), 5.95 (br s, 2H), 3.81 (s, 2H), 2.94 (t, J= 6.9 Hz. 2H), 2.85 (t, J= 6.9 Hz, 2H), 2.51 (s, 1H). 13C NMR (101 MHz, CDCl3) δ 168.83, 161.26, 154.78, 140.02, 136.45, 129.87, 129.49, 128.86, 128.64, 127.62, 126.36, 120.06, 117.65, 53.24, 50.60, 36.36. HRMS (ESI) m / z for [M + H]+calcd for C22H22N2O2, 347.1754; found, 347.1752. HPLC: tR= 2.761 min; purity >99.0%.Synthesis of 6-(2-Chloro-4-((phenethylamino)methyl)phenoxy)nicotinamide (Compound 11).2510712877:vl

[0108] A mixture of powdered molecular sieves (3 A, 0.3 g), 2-phenylethan-l -amine (0.06 mL, 0.43 mmol). Compound 19c (0.10 g, 0.36 mmol), and 1,2-di chloroethane (10 mL) was stirred at room temperature overnight. Sodium triacetoxyborohydride (0.19 g, 0.90 mmol) and acetic acid (0.05 mL, 0.90 mmol) were added and the reaction was stirred for an additional 3 h. The reaction was filtered through a pad of Celite, quenched with saturated aqueous NaHCCh (20 mL), and then extracted with EtOAc (3 x 25 mL). The combined organic extracts were dried (Na2SO4), filtered, and then concentrated in vacuo. The resulting crude material was purified by flash chromatography eluting with 0 - 15% MeOH / EtOAc to afford 0.11 g (79%) of 11 as white solid, mp: 302-303 °C (decomp).!H NMR (600 MHz, DMSO) 5 8.58 (dd, J= 2.4, 0.7 Hz, 1H), 8.30 (dd, J= 8.6, 2.4 Hz, 1H). 8.13 (s, 1H), 7.52 (d, J= 2.0 Hz. 1H), 7.48 (s, 1H), 7.34 (dd, J= 8.3, 2.0 Hz, 1H), 7.29 - 7.24 (m, 3H). 7.23 - 7.20 (m, 2H), 7.19 - 7.16 (m, 1H), 7.14 (d, J= 8.6 Hz, 1H), 3.75 (s, 2H), 2.75 (s, 4H).13C NMR (101 MHz, DMSO) 5 165.79, 164.02, 147.30, 147.28, 140.48, 140.28, 139.62, 129.33, 128.59. 128.21, 127.85, 125.80, 125.42, 123.86, 110.13, 51.67, 50.43, 35.88. HRMS (ESI) m / z for [M + H]+ calcd for C21H20N3O2Cl. 382.1317; found. 382.1297. HPLC: tR= 2.750 min; purity >99.0%.Synthesis of 6-(2-Methoxy-4-((phenethylamino)methyl)phenoxy)nicotinamide (Compound 12).

[0109] A mixture of powdered molecular sieves (3 A, 0.3 g), 2-phenylethan-l -amine (0.06 mL, 0.44 mmol). Compound 19d (0.10 g, 0.37 mmol), and 1,2-dichloroethane (10 mL) was stirred at room temperature overnight. Sodium triacetoxyborohydride (0.19 g, 0.92 mmol) and acetic acid (0.05 mL, 0.92 mmol) were added and the reaction was stirred for an additional 3 h. The reaction was filtered through a pad of Celite, quenched with saturated aqueous NaHCO3 (20 mL), and then extracted with EtOAc (3 x 25 mL). The combined organic extracts were dried (Na2SO4), filtered, and then concentrated in vacuo. The resulting crude material was purified by flash chromatography eluting with 0 - 12% MeOH / EtOAc to afford 0.11 g (82%) of 12 as white solid, mp: 224-225 °C (decomp).1H NMR (600 MHz, DMSO) 5 8.55 (d, J= 2.4 Hz, 1H), 8.21 (dd, J= 8.6, 2.4 Hz, 1H), 8.03 (s, 1H). 7.42 (s, 1H), 7.30 - 7.25 (m, 2H), 7.24 - 7.21 (m, 2H), 7.20 - 7.15 (m, 1H), 7.10 (d, J= 1.9 Hz, 1H), 7.05 (d, J= 8.0 Hz, 1H), 6.98 (d, J= 8.6 Hz, 1H), 6.91 (dd, J= 8.0, 1.9 Hz, 1H), 3.73 (s, 2H), 3.64 (s, 3H), 2.80 - 2.72 (m, 4H).13C NMR (151 MHz, DMSO) 5 165.95, 164.78, 151.08, 147.37, 140.55. 140.03, 139.41, 139.16, 128.62, 128.21. 125.79, 124.72, 122.42, 119.98, 112.52.2610712877:vl109.51, 55.52, 52.61, 50.51, 35.86. HRMS (ESI) m / z for [M + H]+ calcd for C22H23N3O3, 378.1812; found. 378.1837. HPLC: tR= 2.658 min; purity >99.0%.Synthesis of 6-(2-(Benzyloxy)-4-((phenethylamino)methyl)phenoxy)nicotinamide (Compound 13).

[0110] A mixture of powdered molecular sieves (4 A), Compound 19e (0.21 g, 0.634 mmol), phenethyl amine (0.092 g, 0.760 mmol), acetic acid (0.05 mL, 0.951 mmol), THF (1 mL), and MeOH (1 mL) stirred in a vial for 4 h. Sodium triacetoxyborohydride (0.27 g, 1.27 mmol) was added to the reaction and allowed to stir overnight at room temperature. Upon return, reaction was quenched with saturated aqueous NaHCOs (20 mL), then extracted with 10% MeOH / DCM (5 x 10 mL). The combined organic extracts were dried (Na2SO4), filtered, and then concentrated in vacuo. The resulting crude material was purified by flash chromatography eluting with 0 - 0.3% MeOH / DCM to afford 0.11 g (38%) of Compound 13 as a clear, sticky oil. ¹H NMR (600 MHz, MeOD) δ8.57 (dd, J= 2.5, 0.8 Hz, 1H), 8.19 (dd, J= 8.7. 2.5 Hz, 1H), 7.31 - 7.25 (m, 2H), 7.23 - 7.10 (m, 8H), 7.07 - 7.01 (m, 2H), 6.97 (dd, J= 8.1. 1.9 Hz, 1H), 6.93 (dd, J = 8.7. 0.7 Hz. 1H), 4.98 (s, 2H), 3.80 (s, 2H), 2.88 - 2.80 (m, 2H).13C NMR (151 MHz, MeOD) 5 168.29, 165.83, 150.45, 147.20, 141.49, 139.57, 139.14, 137.75, 136.63, 128.35, 128.19, 127.89, 127.43, 126.78, 125.94, 124.37, 122.37, 121.14, 114.35, 109.84, 70.03, 56.07, 52.55, 49.79, 48.17, 47.73, 47.67, 35.22. HRMS (ESI) m / z for [M + H]+calcd for C28H27N3O3, 454.2125; found, 454.2116. HPLC: tR= 3.188; purity = 96%.Synthesis of 6-(4-(((2-Cyclohexylethyl)amino)methyl)-2-methoxyphenoxy)nicotinamide (Compound 14).

[0111] A mixture of powdered molecular sieves (3 A, 0.3 g), 2-cyclohexylethan-l-amine (0.06 g, 0.44 mmol), Compound 19d (0.10 g. 0.37 mmol), and 1.2-dichloroethane (10 mL) was stirred at room temperature overnight. Sodium triacetoxyborohydride (0.19 g, 0.92 mmol) and acetic acid (0.05 mL, 0.92 mmol) were added and the reaction was stirred for an additional 3 h. The reaction was filtered through a pad of Celite, quenched with saturated aqueous NaHCO3 (20 mL), and then extracted with EtOAc (3 x 25 mL). The combined organic extracts were dried (Na2SO4), filtered, and then concentrated in vacuo. The resulting crude material was purified by flash chromatography eluting with 0 - 15% MeOH / EtOAc to afford 0.10 g (72%) of Compound 14 as white solid, mp: 214-215 °C (decomp). ¹H NMR (400 MHz, DMSO) 5 8.54 (d, J = 2.4 Hz, 1H), 8.20 (dd, J = 8.6, 2.5 Hz, 1H), 7.98 (s, 1H).7.42 (s, 1H), 7.12 (d, J = 1.8 Hz, 1H), 7.05 (d, J = 8.0 Hz, 1H), 6.98 (d, J = 8.6 Hz, 1H), 6.922710712877:vl(dd, J= 8.0, 1.8 Hz, 1H), 3.69 (s, 2H), 3.66 (s, 3H), 2.53 (dd, J= 8.6, 5.4 Hz, 2H), 1.70 -1.55 (m, 5H), 1.39 - 1.30 (m, 3H). 1.27 - 1.06 (m, 4H), 0.87 (q, J= 10.4 Hz, 2H).13C NMR (101 MHz, DMSO) 5 165.95, 164.80, 151.06, 147.33, 140.00, 139.59, 139.12, 124.72, 122.37, 119.98, 112.50, 109.51, 55.52, 52.93, 46.39, 37.19, 35.05, 32.99, 26.18, 25.83.HRMS (ESI) m / z for [M + H]+calcd for C22H29N3O3, 384.2282; found, 384.2298. HPLC: tR= 2.967 min; purity >99.0%.Synthesis of 6-(4-((Isopentylamino)methyl)-2-methoxyphenoxy)nicotinamide (Compound 15).

[0112] A mixture of powdered molecular sieves (3 A, 0.3 g), 3-methylbutan-l -amine (0.06 mL, 0.44 mmol), Compound 19d (0.10 g, 0.37 mmol), and 1,2-di chloroethane (10 mL) was stirred at room temperature overnight. Sodium triacetoxyborohydride (0.19 g, 0.92 mmol) and acetic acid (0.05 mL, 0.92 mmol) were added and the reaction was stirred for an additional 3 h. The reaction was filtered through a pad of Celite, quenched with saturated aqueous NaHCO3 (20 mL), and then extracted with EtOAc (3 x 25 mL). The combined organic extracts were dried (Na2SO4), filtered, and then concentrated in vacuo. The resulting crude material was purified by flash chromatography eluting with 0 - 15% MeOH / EtOAc to afford 0.10 g (83%) of Compound 15 as off-white solid, mp: 191-192 °C. 'H NMR (600 MHz, DMSO) 5 8.56 (dd, J= 2.5, 0.8 Hz, 1H), 8.24 (dd, J= 8.6, 2.5 Hz, 1H), 8.09 (s, 1H), 7.42 (s, 1H), 7.13 (d. J= 1.9 Hz. 1H), 7.05 (d, J= 7.9 Hz. 1H), 6.97 (dd, J= 8.6, 0.7 Hz, 1H), 6.92 (dd. J = 8.0. 1.8 Hz, 1H). 3.69 (s. 2H), 3.66 (s. 3H), 2.53 (t, J = 7.4 Hz, 2H). 1.69 - 1.59 (m, J= 6.7 Hz, 1H), 1.35 (q, J= 6.9 Hz, 2H), 0.86 (d, J= 6.6 Hz, 6H).13C NMR (151 MHz, DMSO) 5 165.95, 164.79, 151.06, 147.42, 140.03, 139.45, 139.19, 124.71, 122.39, 120.02, 112.59. 109.48, 55.55, 52.90, 46.90, 38.61, 25.54, 22.66. HRMS (ESI) m / z for [M + H]+ calcd for C19H25N3O3, 344.1969; found, 344.1985. HPLC: tR= 2.554 min; purity >99.0%.Synthesis of 6-(4-(((2,3-Dihydro-17 / -inden-2-yl)amino)methyl)-2-methoxyphenoxy)nicotinamide (Compound 16).

[0113] A mixture of powdered molecular sieves (3 A, 0.6 g), 2,3-dihydro-177-inden-2-amine (0.12 mL, 0.88 mmol), Compound 19d (0.20 g, 0.74 mmol), and 1,2-dichloroethane (20 mL) was stirred at room temperature overnight. Sodium triacetoxyborohydride (0.39 g, 1.84 mmol) and acetic acid (0.11 mL, 1.84 mmol) were added and the reaction was stirred for an additional 3 h. The reaction was filtered through a pad of Celite, quenched with saturated aqueous NaHCO3 (20 mL), and then extracted with EtOAc (3 x 25 mL). The combined organic extracts were dried (Na2SO4), filtered, and then concentrated in vacuo. The resulting2810712877:vlcrude material was purified by flash chromatography eluting with 0 - 20% MeOH / EtOAc to afford 0.24 g (84%) of Compound 16 as solid, mp: 245-246 °C (decomp). 'H NMR (400 MHz, MeOD) 58.56 (dd, J= 2.5, 0.8 Hz, 1H), 8.21 (dd, J= 8.7, 2.5 Hz, 1H), 7.24 - 7.07 (m, 6H), 7.03 (dd, J= 8.1, 1.9 Hz, 1H), 6.91 (dd, J= 8.6, 0.7 Hz, 1H), 3.88 (s, 2H), 3.74 (s, 3H), 3.68 (p, 7.2 Hz. 1H), 3.21 (dd, J= 15.6, 7.4 Hz, 2H), 2.87 (dd, J= 15.6, 7.1 Hz, 2H).13C NMR (151 MHz, MeOD) 5 169.77, 167.16, 153.05, 148.59, 142.54, 142.40, 140.53, 139.27, 127.59. 125.69, 125.54, 123.75, 122.24. 114.44. 110.95, 59.78, 56.28, 52.66. 40.11. HRMS (ESI) m / z for [M + H]+calcd for C23H23N3O3, 390.1812; found, 390.1841. HPLC: tR= 2.711 min; purity 96.0%.Synthesis of 6-(4-Formylphenoxy)nicotinonitrile (Compound 18a).

[0114] A mixture of K.2CO3 (3.40 g, 24.57 mmol), 4-hydroxybenzaldehyde (1.00 g, 8.19 mmol), and 6-fluoronicotinonitrile (1.00 g, 8.19 mmol), and dimethylformamide (15 mL) was heated to 100 °C for 2 h. The reaction was cooled to room temperature and then poured into H2O. The precipitated brown solid was filtered, rinsed with cold H2O, and dried in vacuo to afford 1.51 g (82%) of Compound 18a as solid: 'H NMR (400 MHz, MeOD) 5 9.99 (s. 1H), 8.52 (dt, J= 2.4, 0.7 Hz, 1H), 8.18 (ddd. J= 8.7. 2.4, 0.5 Hz, 1H). 8.03 - 7.97 (m, 2H), 7.41 - 7.33 (m, 2H), 7.23 (dt, J= 8.7, 0.7 Hz, 1H).13C NMR (101 MHz, MeOD) 8 192.83, 166.20, 159.41, 153.12, 144.50, 135.29, 132.55, 123.18, 117.51, 113.75, 106.35.Synthesis of 4-(4-Formylphenoxy)benzonitrile (Compound 18b).

[0115] A mixture of K2CO3 (0.71 g, 5.16 mmol), 4-hydroxybenzaldehyde (0.62 g, 5.08 mmol), and 4-fluorobenzonitrile (0.61 g, 5.08 mmol), and dimethylformamide (20 mL) was heated to 130 °C for 2 h. The reaction was cooled to room temperature and then poured into H2O and extracted with EtOAc (3 x 50 mL). The combined organic extracts were dried (Na2SO4), filtered, and then concentrated in vacuo. The crude material was purified via flash chromatography eluting with 5 - 50% EtOAc / n-hexanes to afford 0.50 g (44%) of Compound 18b as solid: 'H NMR (400 MHz, CDC13) 89.98 (s, 1H), 7.97 - 7.88 (m, 2H), 7.72 - 7.64 (m, 2H), 7.21 - 7.07 (m, 4H).13C NMR (101 MHz, CDC13) 8 190.67, 160.74, 159.73, 134.55. 132.93, 132.24, 119.77, 119.57, 118.50, 107.86.Synthesis of 6-(2-Chloro-4-formylphenoxy)nicotinonitrile (Compound 18c).

[0116] A mixture of K.2CO3 (1.09 g, 7.91 mmol), 3-chloro-4-hydroxybenzaldehyde (0.62 g, 3.95 mmol), and 6-fluoronicotinonitrile (0.48 g, 3.95 mmol), and dimethyl acetamide (5 mL) was heated at 100 °C for 2 h. The reaction w as cooled to room temperature and then poured into H2O and extracted with EtOAc (3 x 25 mL). The combined organic extracts were2910712877:vldried (Na2SO4), filtered, and then concentrated in vacuo. The resulting crude material was purified by flash chromatography eluting with 0 - 20% EtOAc / n-hexanes to afford 0.48 g (64%) of Compound 18c as white solid: 'H NMR (600 MHz, CDCh) 69.99 (s, 1H), 8.40 (d, J = 2.3 Hz, 1H), 8.02 (d, J= 1.9 Hz, 1H), 8.01 (dd, J= 8.6, 2.3 Hz, 1H), 7.88 (dd, J = 8.3, 1.9 Hz, 1H), 7.40 (d, J= 8.3 Hz, 1H), 7.19 (d, J = 8.6 Hz, 1H). 13C NMR (151 MHz, CDCl3) δ 189.73, 164.26, 153.55, 151.96, 142.85, 135.15, 131.93, 129.55, 128.84, 124.69, 116.49. 112.15, 105.35.Synthesis of 6-(4-Formyl-2-methoxyphenoxy)nicotinonitrile (Compound 18d).

[0117] A mixture of K2CO3 (0.91 g, 6.57 mmol), vanillin (4-hydroxy-3-methoxybenzaldehyde) (0.5 g, 3.29 mmol), and 6-fluoronicotinonitrile (0.41 g, 3.29 mmol), and dimethylacetamide (2 mL) was heated at 100 °C for 2 h. The reaction was cooled to room temperature and then poured into H2O and extracted with EtO Ac (3 x 25 mL). The combined organic extracts were dried (Na2SO4), filtered, and then concentrated in vacuo. The resulting crude material was purified by flash chromatography eluting with 0 - 100% EtOAc / n-hexanes to afford 0.70 g (84%) of Compound 18d as solid: 'H NMR (400 MHz, CDC13) 59.98 (s, 1H), 8.39 (d, J = 2.3 Hz, 1H), 7.95 (ddd, J = 8.6, 2.4, 0.9 Hz, 1H), 7.58 -7.50 (m, 2H), 7.35 - 7.28 (m, 1H), 7.11 (d, J = 8.6 Hz, 1H), 3.81 (s, 3H). 13C NMR (101 MHz, CDCl3) δ 191.02, 164.98, 152.33. 152.03, 146.66, 142.39, 135.27, 125.16, 123.48, 116.78. 111.77, 111.26, 104.62, 56.15.Synthesis of 6-(4-Formylphenoxy)nicotinamide (Compound 19a).

[0118] A mixture of Compound 18a (4.83 g, 21.54 mmol) and K2CO3 (5.96 g, 43.10 mmol) in DMSO (50 mL) was cooled to 10 °C. A solution of aqueous 30% H2O2 (7.34 mL, 94.79 mmol) was added in a dropwise manner and the resulting mixture was stirred and allowed warm to room temperature over 2 h. The reaction was poured into H2O and extracted with EtOAc (3 x 25 mL). The combined organic extracts were dried (Na2SO4), filtered, and then concentrated in vacuo. The resulting crude material was purified by flash chromatography eluting with 0 - 80% EtOAc / n-hexanes to afford 4.05 g (78%) of Compound 19a as solid: 'H NMR (400 MHz, DMSO) 5 10.00 (s, 1H), 8.66 (dt, J= 2.5, 0.7 Hz, 1H), 8.32 (ddd, J= 8.6, 2.5, 0.6 Hz, 1H), 8.08 (s, 1H), 8.03 - 7.94 (m, 2H), 7.53 (s, 1H), 7.41 - 7.35 (m, 2H), 7.22 (dt, J= 8.5, 0.7 Hz, 1H).13C NMR (101 MHz, DMSO) 5 191.89, 165.69, 163.79, 158.50, 147.54, 139.87, 132.92, 131.48, 126.09, 121.60, 111.60.Synthesis of 4-(4-Formylphenoxy)benzamide (Compound 19b).3010712877:vl

[0119] A mixture of Compound 18b (0.38 g, 1.72 mmol) and K2CO3 (0.12 g, 0.86 mmol) in DMSO (1.6 mL) was cooled to 10 °C. A solution of aqueous 30% H2O2 (0.19 mL, 1.89 mmol) was added in a dropwise manner and the resulting mixture was stirred and allowed warm to room temperature over 2 h. The reaction was poured into H2O and extracted with EtOAc (3 x 30 mL). The combined organic extracts were dried (Na₂SO₄), filtered, and then concentrated in vacuo to afford 0.37 g (90%) of Compound 19b as shiny white solid:1H NMR (400 MHz, DMSO) δ 9.95 (s, 1H), 8.02 - 7.90 (m, 5H), 7.38 (s, 1H), 7.19 (dd, J = 8.5, 4.4Hz, 4H).13C NMR (101 MHz, DMSO) 5 191.60, 167.01, 161.49, 157.40, 132.05, 131.79, 130.58, 129.93, 119.23, 118.36.Synthesis of 6-(2-Chloro-4-formylphenoxy)nicotinamide (Compound 19c).

[0120] A mixture of Compound 18c (0.37 g, 1.42 mmol) and K.2CO3 (0.10 g. 0.71 mmol) in DMSO (1.3 mL) was cooled to 0 °C. A solution of aqueous 30% H2O2 (0.16 mL, 1.56 mmol) was added in a dropwise manner and the resulting mixture was stirred and allowed warm to room temperature over 2 h. The reaction was poured into H2O and extracted with EtOAc (3 x 25 mL). The combined organic extracts were dried (Na2SO4), filtered, and then concentrated in vacuo to afford 0.24 g (62%) of Compound 19c as white solid: 'H NMR (600 MHz, DMSO) 5 10.00 (s, 1H), 8.58 (dd, J= 2.5, 0.7 Hz, 1H), 8.34 (dd, J= 8.6, 2.4 Hz, 1H), 8.14 (d, J = 2.0 Hz, 1H), 8.07 (s, 1H), 7.96 (dd, J= 8.3, 2.0 Hz, 1H), 7.57 (d, J = 8.3 Hz, 1H), 7.53 (s, 1H), 7.29 (dd, J= 8.6, 0.7 Hz, 1H).13C NMR (151 MHz, DMSO) 5 191.15, 165.64, 163.40, 153.75, 147.32, 140.03, 134.55, 131.53, 129.52, 127.19, 126.13, 124.86, 110.76.Synthesis of 6-(4-Formyl-2-methoxyphenoxy)nicotinamide (Compound 19d).

[0121] A mixture of Compound 18d (0.70 g, 2.75 mmol) and K.2CO3 (0.76 g. 5.51 mmol) in DMSO (10 mL) was cooled to 10 °C. A solution of aqueous 30% H2O2 (0.94 mL, 12.11 mmol) was added in a dropwise manner and the resulting mixture was stirred and allowed warm to room temperature over 2 h. The reaction was poured into H2O and extracted with EtOAc (3 x 25 mL). The combined organic extracts were dried (Na2SO4), filtered, and then concentrated in vacuo. The resulting crude material was purified by flash chromatography eluting with 0 - 15% MeOH / CH₂Cl₂ to afford 0.65 g (87%) of Compound 19d as white solid: 'H NMR (400 MHz, DMSO) 59.99 (s, 1H). 8.54 (dd. J = 2.5, 0.7 Hz, 1H), 8.27 (dd, J = 8.6, 2.4 Hz, 1H), 8.03 (s, 1H), 7.66 - 7.58 (m, 2H), 7.48 (s, 1H), 7.41 (d, J = 7.8 Hz, 1H), 7.15 (dd, J = 8.7, 0.8 Hz, 1H), 3.77 (s, 3H).13C NMR (101 MHz, DMSO) 53110712877:vl191.99, 165.81, 164.08, 152.02, 147.28, 146.84, 139.51, 134.48, 125.40, 124.03, 123.61, 112.14. 110.12, 55.92.Synthesis of 6-(2-(Benzyloxy)-4-formylphenoxy)nicotinamide (Compound 19e).

[0122] A mixture of K2CO3 (0.83 g, 6 mmol), 3-(benzyloxy)-4-hydroxybenzaldehyde (0.69 g, 3 mmol), and 6-chloronicotinamide (0.31 g, 2 mmol), and DMF (20 mL) was heated at 100 °C for 2 h. The reaction was cooled to room temperature and then poured into H2O and extracted with EtOAc (3 x 25 mL). The combined organic extracts were dried (Na₂SO₄), filtered, and then concentrated in vacuo. The resulting crude material was purified by flash chromatography eluting with 15 - 60% EtO Ac / n-hexanes to afford 0.23 g (33%) of Compound 19e as an orange oil:1H NMR (400 MHz, DMSO) δ 9.99 (s, 1H), 8.59 (d, J= 2.4 Hz, 1H), 8.28 (dd, J= 8.6, 2.5 Hz, 1H), 7.71 (d, J= 1.8 Hz, 1H), 7.66 (dd, J= 7.9, 1.8 Hz, 1H), 7.48 (d, J= 8.1 Hz, 2H), 7.24 (m, 3H), 7.18 (d, J= 8.6 Hz, 1H), 7.04 (dd, J = 6.6, 2.9 Hz, 2H), 5.15 (s, 2H).General procedure for hydrochloride salt formation.

[0123] HC1 (IM in Et20, 1 equiv) was added to a solution of the appropriate free base in MeOH. Once in solution, the MeOH was removed in vacuo and Et20 was added to the crude residue. The resulting precipitate was collected by filtration and dried under vacuum to afford the corresponding hydrochloride salt.General procedure for oxalate salt formation.

[0124] Oxalic acid (1 equiv) was added to a solution of the appropriate free base in MeOH. Once in solution, the MeOH was removed in vacuo and Et20 was added to the crude residue. The resulting precipitate was collected by filtration and dried under vacuum to afford the corresponding oxalate salt.Cell Lines and Cell Culture.

[0125] The cAMP Hunter™ CHO-K1 stably expressing the human p. opioid receptor (OPRM1), K opioid receptor (OPRK1), the human 8 opioid receptor (OPRD1), and nociceptin receptor (OPRL1) were purchased from Eurofins DiscoverX (Fremont, CA, USA) and maintained in F-12 media supplemented with 10% fetal bovine serum (Life Technologies, Grand Island, NY, USA), 1% penicillin / streptomycin / L-glutamine (Life Technologies), and 800 pg / mL Geneticin (Minis Bio, Madison, WI, USA). HEK293T cells (Thermo Fisher Scientific) were cultured in Dulbecco's modified Eagle’s medium (DMEM, Coming) containing 10% fetal bovine serum and 1% penicillin / streptomycin. All cells were grown at 37 °C and 5% CO2 in a humidified incubator.3210712877:vlForskolin-induced cAMP accumulation assay.

[0126] Briefly, cells were plated in a 384-well white tissue culture plate (10,000 cells / well) and incubated overnight at 37 °C. Compounds were first dissolved in DMSO to form stock solutions (5 mM concentration), and then 9 doses of 100X solutions were prepared by serial dilution with DMSO. Then 5X solutions were prepared by diluting 100X solutions with assay buffer consisting of Hank’s Buffered Salt Solution and HEPES, with (agonist) or without (antagonist) forskolin. In the antagonist assays, cells were pretreated with compounds for 15 min at 37 °C followed by 30 min incubation at 37 °C with selected agonists at their EC₅₀ or EC90 dose. The HitHunter cAMP Assay for Small Molecules by Eurofins DiscoverX (Fremont, CA, USA) was then used according to manufacturer’s directions and the BioTek Synergy Hl hybrid and Cytation 5 plate readers (BioTek, Winooski, VT, USA) and Gen5 Software version 2.01 (BioTek, Winooski, VT, USA) were used to quantify luminescence.Example 2. Evaluation of Opioid Antagonism.

[0127] Compounds were characterized at opioid receptors by the cAMP accumulation assay (Table 1).Table 1. Evaluation of Opioid AntagonismMOR KOR DOR% Imax pIC₅₀ ± pIC₅₀ % Imax Cmpd pICso ± IC50 + IC50 CN SEM / Olmax □_ IC50 + S ± SEMSEMa’b(nM) (nM) SEMe(nM) MPO SEMCa,d a,f SEMC eNLX 7.5 ± 120.5 ± 6.8 ± 73.2 ± 6.9 ± 110.6 ±33 170 140 5.4 0.1 2.4 0.1 11.7 0.1 4.6 NTX 8.1 ± 103.4 ± 8.3 ± 41.3 ± 6.6 ± 99.9 ±7.5 5.3 280 5.3 0 1 04 0 1 6.8 0 1 068.7 ± 128.8 ± 8.1 ± 16.8 ± 6.9 ± 90.5 ± NLM 2.1 7.6 140 5.20.1 7.1 0.2 27 0.1 12.4 DPN 8.6 ± 88.9 ± 8.7 ± 4.8 ±26 22 < 5 0 4 1 0.1 3.6 0.1 1 8 10.000 LY255.582 8.7 ± 165.1 ± 8.3 ± 93.5 ± 8.4 ± 155.4 ±2.0 5.4 3.7 4.1 0.1 23.5 0.1 1 8 0.1 13.262 ± 1192 ±NLX-M 660 N / D N / D N / D N / D N / D N / D 4.70.1 1.26.41 130.5 ± 6.21 103.4 ± Naloxegol 410 N / D N / D N / D 690 3.30.5 7.8 0.1 7.9 Naldemedine 8.5 ± 135.9 ±3.5 N / D N / D N / D N / D N / D N / D 1.8 0.1 8.07.9 ± 153.8 ±1 22.9 N / D N / D N / D N / D N / D N / D 2.6 0.8 6.37.4 ± 188.8 ±2 40.1 N / D N / D N / D N / D N / D N / D 2.90.2 38.19.0 ± 149.3 ± 8.0 ± 106.1 ± 8.8 ± 229.9 ± 9 1 0 10 1 7 450.1 11.1 0.2 30 0.1 55.510 8.7 ± 147.7 ± 8.0 ± 109.6 ± 8.5 ± 189.5 ±2.1 10 3.0 4.1 0.1 5.7 0.1 63 0.1 16.611 8.9 ± 152.5 ± 7.7 ± 105.4 ± 8.7 ± 158.1 ±1.2 19 2.1 4.1 0.1 13.8 0.1 75 0.1 7.59.2 ± 156.1 ± 7.3 ± 106.1 ± 9.2 ± 156.8 ± 12 0.65 46 058 4.20.2 16.4 0.3 88 0.1 5.23310712877:vl8.5 ± 134.1 ± 8.4 ± 152.3 ± 13 0.1 3.2 18.3 N / D N / D N D 0.2 3.9 14.0 3.2 14 8.5 ± 3 0 146.1 ± 7.7 ± 22 120.3 ± 8.8 ± 1 5 173.6 ± 370.1 12.5 0.1 17.1 0.1 18.8 8.7 ± 146.4 ± 7.7 ± 8.3 ± 187.0 ± 15 2.0 22 115.5 ± 5.2 4.20.2 11.8 0.1 5 1 0.2 19.9 94 ± 183.7 ± 8.0 ± 111 3 ± 92 ± 1678 ± 16 0.1 0.39 41.7 0.2 10 15.0 0.1 062 0.6 4.6 EW-745 8.4 4.0 3634 N / D N / D N / D N / D N / D N / D N / D EW-738 8.5 3.2 2893 N / D N / D N / D N / D N / D N / D N / D 9.0 ± 2453EW-725 1.0 N / D N / D N / D N / D N / D N / D N / D 0.1 ± 15.29.3 ± 2308EW-710 0.50 N / D N / D N / D N / D N / D N / D N / D 0.3 ± 36.189 ± 146.8JWP-366 1.3 N / D N / D N / D N / D N / D N / D N / D 0.3 ± 15.36.5 ± 1409JWP-271 320 N / D N / D N / D N / D N / D N / D N / D 0.2 ± 21.764 ± 1381JWP-272 400 N / D N / D N / D N / D N / D N / D N / D 0.1 ± 6.87.6 ± 1650JWP-231 25 N / D N / D N / D N / D N / D N / D N / D 0.3 ± 31.48.2 ± 1669JWP-230 6.3 N / D N / D N / D N / D N / D N / D N / D0.3 ± 36.4aValues are expressed as the mean ± standard error of the mean (SEM) of at least three independent experiments.bAntagonist potency (IC50) determined versus EC90 of fentanylcDegree of antagonism (Imax) normalized to naltrexonedAntagonist potency (IC50) determined versus EC90 of U50.488eDegree of antagonism (Imax) normalized to wor-BNIfAntagonist potency (IC50) determined versus ECso of SNC-80eCalculated using CDD Vault N / D = not determinedEW-745 EW-738EW-7253410712877:vlJWP-271JWP-272JWP-231JWP-230

[0128] As expected, Compounds 9 - 16 were found to possess antagonist activity greater than NLX (pICso > 7.5). Aminobenzyloxyarylamides Compounds 9, 12, and 16 were found to be highly potent p-opioid receptor (MOR) antagonists (pICso > 9.0). Interestingly, Compounds 9 - 16 were found to be more efficacious as antagonists than NLX (Imax > 121%). Not wishing to be bound by theory, the reasons for the increased efficacy may relate to the assay conditions used to profile antagonist activity (cAMP versus [35S]GTPgS, short incubation time, and EC90 concentration of fentanyl). Some structure-activity relationships at MORs were readily observable. The pyridine ring in Compound 9 could be readily substituted with a benzene ring (Compound 10) with little reduction in MOR antagonist activity (Compound 10: pICso = 8.7 vs. Compound 9: pICso = 9.0). Substitution of a chlorine 3510712877:vl(Compound 11) and a methoxy group (Compound 12) in the aryloxy ring were also well tolerated (Compound 11: pICso = 8.9 and Compound 12: pICso = 9.2 vs. Compound 9: pICso = 9.0). However, the conversion of the methyl group in Compound 12 to a benzyloxy group (Compound 13) resulted in a 5-fold loss of activity (Compound 13: pICso = 8.5 vs.Compound 12: pICso = 9.2). Finally, the phenethyl substituent could also be substituted with a cyclohexylethyl (Compound 14) or 3-methylbutanyl (Compound 15) group with a loss in activity at MORs (Compound 14: pICso = 8.5 and Compound 15: pICso = 8.7 vs. Compound 12: pICso = 9.2). Conformational constraint of the phenethyl group into a 2-indane (Compound 16) resulted in an increase in MOR antagonist activity (Compound 16: pICso = 9.4 vs. Compound 12: pICso = 9.2). Compounds EW-745, EW-738, EW-725, and EW-710 exhibited full antagonism, with a normalized Imax of > 200%.

[0129] Schild analysis was done by generating fentanyl dose-response curves in the absence and presence of three doses of test compounds. Data were analyzed by nonlinear regression with the Gaddum / Schild ECso shift function in GraphPad Prism. Compounds with Schild slope close to one were considered competitive, and pA2 values were calculated (constraining both HillSlope and SchildSlope to 1). The equilibrium dissociation constant (Ke) values were calculated as well using the formula: Ke= [L] / [(A7A)-1 ]; [L] is the concentration of antagonist, and A’ and A are the ECso of fentanyl in the presence or absence of a single dose of the antagonist, respectively.

[0130] A Schild analysis was conducted on antagonists using a full dose-response curve of fentanyl in the absence or presence of three concentrations of antagonist (1, 10, and 100 nanomoles per liter). The antagonists w ere NTX (FIG. 1A), NLX (FIG. IB), and Compound 9 (FIG. 1C). The Schild slope was determined to not significantly deviate from 1, indicating competitive antagonism (FIGS. ID and IE). The pA values (the concentrations of antagonist required to have a 2-fold increase in the concentration of agonist to produce a selected effect) was found to be 10.66 compared to 8.48 for NLX and 9.47 for naltrexone (NTX). The values are expressed as the mean ± SEM of at least three independent experiments.

[0131] Compounds 9 - 16 were then evaluated for their antagonism at KORs and DORs (Table 1). Compounds 9 - 16 were found to be KOR antagonists more potent and efficacious than NLX (pICso > 6.8 and Imax > 73.2%) and more efficacious than nalmefene (NLM) (Imax = 16.8%). Aminobenzyloxyarylamides 9 - 16 were also found to be DOR antagonists more potent than NLX, NTX, and NLM (pICso > 8.3 vs. pICso = 6.6 - 6.9).3610712877:vlCollectively, aminobenzyloxyarylamides 9 - 16 can be characterized as potent but non-selective full opioid antagonists.Bioluminescence resonance energy transfer (BRET) assay.

[0132] The antagonist activity of Compound 12 on G-protein and P-arrestin2 recruitment at MOR was evaluated in HEK293T cells using BRET2 and BRET1 assays, respectively. A 1:2 ratio of plasmids to Transit 2020 (Minis Biosciences) was used for transfection in 10 cm dishes. HEK293T cells were transfected with 500 ng MOR, 500 ng Gail-RLuc8, 500 ng G|33, and 500 ng GFP2-G / 9 for BRET2, while 500 ng MOR-RLuc8 and 2.5 pg mVenus-0-arrestin2 were used for BRET1 assessment. Sixteen hours posttransfection, cells were plated in poly-L-lysine-coated 96-well white clear-bottom plates at a density of 40,000-50,000 cells per well in 200 pL of plating medium (DMEM with 1% dialyzed fetal bovine serum). The following day, cells were washed once with 60 pL of drug buffer (1 x HBSS and 20 mM HEPES, pH 7.4) and then incubated with 60 pL of RLuc substrate, with coelenterazine 400a for BRET2 and coelenterazine h for BRET 1. This was followed by the addition of 30 pL of 3× final concentrations of the antagonist, prepared in drug buffer supplemented with 0.3% (w / v) BSA. The cells were then incubated in the dark at room temperature for 15 minutes, followed by the addition of 30 pL of 4× EC₅₀ DAMGO prepared in drug buffer containing 0.3% (w / v) BSA, and further incubated for 5 minutes. With the Mithras LB 940 multimode microplate reader (Berthold Technologies), GFP2 emission at 510 nm and RLuc8-coelenterazine 400a emission at 395 nm were detected for Got protein activation, while mVenus emission at 485 nm and coelenterazine h-RLuc8 emission at 530 nm were measured for P-arrestin2 recruitment, with a 1 -second read per well. The GFP to Rluc8 ratio and mVenus to Rluc8 ratio were calculated and plotted against drug concentration, normalized to the percentage of NLX inhibition, and analysed using log (inhibitor) vs. response in GraphPad Prism to detect the examined antagonist’s potency and efficacy.Example 3. BRET Assays of Antagonism

[0133] BRET assays were used to directly measure the antagonism of DAMGO-mediated Gil protein heterotrimer dissociation (FIG. 2A) or p-arrestin2 recruitment (FIG. 2B). NTX and Compound 12 were found to be balanced and full antagonists compared to NLX for MOR-Gil signaling (NTX: Imax= 97.8 ± 1.5% vs. 12: Imax= 111.1 ± 1.9%). In addition, NTX and Compound 12 also antagonized DAMGO-induced p-arrestin2 activation like NLX (naltrexone: Imax = 94.4 ± 1.5%; Compound 12: Imax= 100.1 ± 3.3%) (Table 2).3710712877:vlTable 2. Reversal of DAMGO-induced activation of the MOR by NLX, NTX, and Compound 12.MOR-Gil MOR-|3-arrestin2Cmpd plCjo ± SEM IC50 (nM) %Imax ± pICjo ± SEM IC50 (nM) %Imax ± SEM SEM NLX 7.9 ± 0.05 13.1 99.8 ± 1.6 8.2 ± 0.05 6.2 99.8 ± 1.5 NTX 84 ± 005 3 8 978 ± 1 5 89 ± 006 1 4 944 ± 1 512 9.3 ± 0.05 0.5 111.1 ± 1.9 10.0 ± 0.1 0.0 100.1 ± 3.3

[0134] Values are expressed as the mean ± SEM of three biological replicates.Antagonist potency (IC50) determined versus ECso of DAMGO. Degree of antagonism (Imax) normalized to NLX.Animals.

[0135] All procedures were in accordance with the NIH Guide for the Care and Use of Laboratory Animals (8thedition) and approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Kentucky. For antinociception experiments, male and female Fl hybrid mice (n=31) arrived in the laboratory between 8 - 10 weeks of age (Taconic Biosciences, Cambridge City, IN, USA). For locomotor and ventilation experiments, male and female Sprague-Dawley rats (n=8 per experiment) arrived in the laboratory between 50 - 55 days of age (Envigo-Harlan, Indianapolis, IN, USA). Both mice and rats were maintained on an automated 12h:12h light-dark cycle and individually housed with ad libitum access to both food and water in the home cage. All testing occurred during the early portion of the light phase.Drugs for in Vivo Experiments.

[0136] Fentanyl HC1 (National Institute on Drug Abuse Drug Supply Program) was dissolved in 0.9% NaCl saline (1 mg / kg) and delivered via subcutaneous (s.c.) injection. Carfentanil (100 g / mL in MeOH) (Cayman Chemical, Ann Arbor, MI, USA) was diluted in saline and delivered via s.c. injection. Compound 9 was dissolved in 10% Captisol™ and was administered via s.c. injection. Doses for Compound 9 and carfentanil were expressed as free base weight, while doses for fentanyl were expressed as salt weight. All injections were 0.1 mL / kg of body weight. All doses were delivered in counterbalanced order.Apparatus for in Vivo Experiments.

[0137] Antinociception was recorded via hotplate (11 x 11 x 15; Columbus Instruments, Columbus, OH, USA) with paw lick latency normalized to show maximal possible of effect (MPE). Locomotor activity was recorded via automated horizontal activity monitors (24 x 24 x 30; AccuScan Instruments Inc., Columbus, OH, USA) with distance traveled measured in 5-min intervals. Ventilatory effort was measured via whole body 3810712877:vlplethysmography chambers (WBP) (Data Science International, New Brighton, MN, USA) with minute ventilation, breaths per minute, and tidal volume recorded in 5-min intervals. Sensitive pressure transducers were attached to a data acquisition system to record pressure oscillations associated with rodent respiration. Airflow through the plethysmograph (~3L / min) was maintained constant using appropriate flow controllers to prevent CO2 accumulation and re-breathing of exhaled gas.General procedure for Antinociception.

[0138] Mice were habituated to the testing room for one hour, and then placed on the 55 °C hotplate where baseline measurements were taken. To measure paw lick latency, a timer was started when the mouse first touched the hotplate and was stopped with the lick of one paw, either front or hind. Animals were then given fentanyl (1 mg / kg, s.c.) and 10 min later were placed on the hotplate and the first post-injection measurement was taken. If the animal did not respond to the hotplate after 30 sec, the animal was removed from the hotplate and a paw lick latency of 30 sec was recorded. Five min later, the lowest dose of test compound was administered and then 5 min later mice were placed back onto the hotplate and the second post-injection measurement was taken. This test procedure was repeated 4 subsequent times, with the test dose increasing logarithmically each time until either the maximum dose (1 mg / kg, s.c.) was reached or until the paw-lick latency was reduced to the duration observed during the initial baseline test, i.e., full reversal of fentanyl-induced antinociception was reached. After the final post-injection measurement, animals were returned to their home cage and monitored for recovery.Example 4. Antinociception Testing.

[0139] The ability of several aminobenzyloxy arylamides to inhibit the antinociceptive effects of fentanyl (1 mg / kg, i.p.) were evaluated when tested in the hot plate (HP) assay using C57 / 129S Fl hybrid mice. The experimental design dosing schedule is provided in FIG. 3 A. The Nociceptive dose-response curves for NTX (AD50 = 0.002422 mg / kg, n = 13, R2= 0.7255), Compound 11 (AD50 = 2.955e-005 mg / kg, n = 9, R2= 0.2347), Compound 12 (AD50 = 2.924e-005 mg / kg, n = 10, R2= 0.207). Compound 15 (AD50 = 3.933e+023 mg / kg, n = 21, R2= n / a), Compound 16 (AD50 = 4.186e-005 mg / kg, n = 6, R2= 0.5423). and Compound 9 (AD50 = 0.01572 mg / kg, n = 6, R2= 0.7159) (FIG. 3B). Though none of the tested compounds showed full reversal comparable to NLX, several analogues including Compound 9 were found to significantly inhibit the antinociceptive activity of fentanyl. Compound 9 was found to provide the best-balanced properties: good in vitro potency (pICso3910712877:vl= 9.0) and good oxidative metabolic stability in mouse [ti 2= 62.9 min; CLint= 22.05 mL / min / mg] (data not shown).General procedure for Chronic Withdrawal Testing.

[0140] Prior to testing, mice were given daily injections of fentanyl (1 mg / kg) for 10 consecutive days. On the day of testing, baseline hotplate measurements were taken prior to fentanyl administration (1 mg / kg, intraperitoneal injection (i.p.)). Mice dosed with fentanyl were allowed 10 minutes before being given either NLX (10 mg / kg), the peripherally restricted antagonist naloxone methiodide (NLX-M, 10 mg / kg), Compound 9 (1 mg / kg or 10 mg / kg), or saline. Each mouse was subjected to a 30 minute open field test, in which two researchers blinded to the compounds observed and noted any instances of the following symptoms: jumping, grooming, rearing, urinations, fecal boli. persistent trembling, ptosis, hyperirritability, writhing, wet dog shakes, teeth chattering, hunched posture, and piloerection. Each withdrawal behavior was scored based on how many times the behavior occurred during the open field session, w ith 0 times resulting in a score of 0, 1-3 times having a score of 1, 4-6 times scored a 2, and 6+ times was given a score of 3. The scores of each symptom count were added together, resulting in a Cumulative Withdrawal Score. A secondary Cumulative Withdrawal Score was considered as well, in case symptoms that relate to breathing issues confounded the withdrawal. This secondary withdrawal score included scores only from the following behaviors: Urinations, Fecal Boli, Rearing, Grooming, Ptosis, and Hyperirritability.Example 5. Withdrawal Testing.

[0141] To assess efficacy in opioid rescue, the efficacy of Compound 9 was tested in vivo for precipitation of opioid w ithdraw al. Administration of NLX or other opioid antagonist to an opioid-dependent animal can precipitate a moderate-to-severe w ithdraw al syndrome. To further understand the pharmacology of Compound 9, we evaluated its ability to precipitate w ithdraw al in fentanyl -dependent mice. FIG. 4A provides a timeline of the chronic withdrawal testing and observed withdrawal behaviors. Mice chronically dosed w ith 1 mg / kg of fentanyl were allowed 10 minutes before being given 10 mg / kg of either NLX, the peripherally restricted antagonist naloxone methiodide (NLX-M). Compound 9, or saline. Each mouse was subjected to a 30 minute open field test, in which two blinded researchers observed and noted any instances of the following signs: jumping, grooming, rearing, fecal boli, urinations, persistent trembling, ptosis, and piloerection. The overall withdraw al score for NLX (10 mg / kg). NLX-M (10 mg / kg). and Compound 9 (1 and 10 mg / kg) versus a saline control are illustrated in FIG. 4B. The asterisks indicated * Significant between-group 4010712877:vldifference using Chi-Squared analysis, p<0.05. The withdrawal score without metrics potentially impacted by breathing are shown in FIG. 4C for the NLX. NLX-M, and Compound 9 versus a saline control. As expected, NLX produced significant withdrawal symptoms in the fentanyl-dependent mice. In contrast, NLX-M precipitated little withdrawal symptoms. Like NLX, Compound 9 (10 mg / kg) was found to produce significant withdrawal compared to saline. However, a lower dose of Compound 9 (1 mg / kg) produced a similar level of withdrawal compared to NLX-M.

[0142] Given the unusual in vivo profile of Compound 9 as a rescue agent for ultrapotent synthetic opioids (UPSO) exposure, it was assessed for signs of w ithdraw al when used as a rescue agent following an acute challenge with carfentanil (15 mg / kg, s.c.). When rescued with Compound 9 (1.7 mg / kg. s.c ), rats displayed a return to normal levels of horizontal activity, rearing and grooming, but showed no evidence of withdraw al based on wet dog shakes, piloerection or ptosis (results not shown).General procedure for Locomotion and Respiration.

[0143] In these experiments, rats (n=8 per experiment) were first given 2 days of habituation to the locomotor and WBP chambers prior to the first day of drug testing. On each habituation day, rats were brought to the test room and placed in the locomotor apparatus for 15 min, follow ed immediately by placement into the WBP chamber for 30 min. In the first experiment, following habituation, four rats were randomly assigned to a fentanyl treatment group and four rats were randomly assigned to the saline treatment group. On each test day, rats were given either fentanyl (200 pg / kg, s.c.) or 0.9% NaCl (saline, s.c.), followed 15 min later by one of varying doses of Compound 9 (0.001-1.0 mg / kg, s.c.) or the vehicle. Immediately following the second injection, rats were placed into the locomotor chamber for 15 min to measure distance traveled (cm). Following the locomotor session, rats were given a righting reflex (RR) test. For this test, rats were placed into a supine position within the locomotor chamber by the experimenter and time to return to the upright position was recorded, with a maximum cutoff of 30 sec. This test was repeated three times before the rat was then placed into the WBP chamber where their minute ventilation, breathing rate, and tidal volume were recorded for 30 min. Doses of Compound 9 were given in counterbalanced order, with a minimum of 2 days separating each test day. After all doses of Compound 9 were tested, group assignment for each rat w as reversed such that fentanyl rats were reassigned to the saline group, while saline rats were assigned to the fentanyl group. As before,4110712877:vleach dose of Compound 9 or saline was tested in counterbalanced order, thus yielding a final n=8 for each group (fentanyl or saline) and Compound 9 dose condition.

[0144] In the second experiment, similar procedures were used except that rats were assigned to groups that received either carfentanil (15 µg / kg, s.c.) or 15% MeOH in saline and the dose range of Compound 9 was slightly higher (0.01-3.0 mg / kg, s.c.); in addition, a second 15-min locomotor test session was performed immediately after the WBP test session.Example 6. Locomotion and Respiration Testing.

[0145] The ability of Compound 9 to rescue the locomotor and ventilatory depressant effects of fentanyl and carfentanil, respectively was evaluated. A timeline for the locomotor and ventilation assays are shown in FIGS. 5A and 7A. Rats were first challenged with either fentanyl (200 µg / kg, s.c.) or carfentanil (15 µg / kg, s.c.) or saline, followed by one of varying doses of Compound 9 (0.003 - 2 mg / kg, s.c.) or vehicle. In contrast to NLX, NTX and NLM, Compound 9 did not produce a dose-dependent reversal of either fentanyl-induced or carfentanil-induced motoric incapacitation (FIGS. 5B and 7B); however, this was likely due to incomplete absorption at the time of testing because a follow-up experiment showed a complete reversal when locomotion was measured after the respiration assay (FIG. 7C). Unexpectedly, not only was the motor incapacitation completely reversed in the postplethysmography test, Compound 9 produced a hyperactive response in carfentanil-treated rats compared to saline controls (FIG. 7C). Additionally, Compound 9 was able to produce a full reversal of the righting reflex in rats treated with either carfentanil or fentanyl, suggesting a reversal of the motor rigidity (catalepsy) associated with WCS, though this effect could also be explained by fentanyl-induced sedation (FIGS. 5C, 7D, and 7E). More surprisingly, in contrast to standard antagonists, Compound 9 not only reversed fentanyl-induced ventilatory depression, but stimulated ventilation above control levels in rats that had received the fentanyl attack; no respiratory effect of Compound 9 alone was observed in saline controls within the dose range tested (see FIGS. 6A - 6C). Similar results with Compound 9 were seen when tested against the highly potent synthetic opioid, carfentanil, i.e., Compound 9 reversed carfentanil-induced ventilatory depression factors above control levels (FIGS. 8A -8C). For the data shown in FIGS. 6A - 8C, the # symbol indicates significant between-subject difference from saline group.

[0146] The ability of NLX-M to reverse fentanyl-induced respiratory depression in rats was investigated. Quaternary nitrogen derivatives, such as NLX-M, are thought not to readily cross the blood-brain barrier. Like Compound 9, NLX-M (5 mg / kg, i.v.) was able to4210712877:vlreverse fentanyl-induced respiratory depression (FIG. 9). Collectively, these results suggest that Compound 9, or an active metabolite, may have entered the brain in small trace amounts that were sufficient to precipitate centrally mediated withdrawal signs due to its high activity at MORs in mice. Unexpectedly, Compound 9 has limited brain penetrance and is more potent in vitro than NLX-M (pIC₅₀ of 9.0 versus 6.2 as shown in Table 1).Example 7. Evaluation of In vitro Metabolism.

[0147] The metabolic stability of selected analogues of aminobenzyloxyarylamides in mouse and human microsomes were evaluated (Table 3).Table 3. Evaluation of metabolic stability in human liver microsomes (HLM) and mouse liver microsomes (MLM) and inhibition of selected CYP enzymes in HLMs at 10 pM.% Inhibition at 10 mMCmpd HLMti / 2 MLM ti21A2 2C9 2C19 2D6 3A4 (min) (min)NLX > 145 14.4 2.0 0.0 0.0 17.9 1.4 NTX > 145 57.4 1.0 0.0 0.0 36.7 0.0 NLM > 145 87.1 0.0 0.0 0.0 68.2 0.2 DPN 4.8 < 2.5 6.0 2.3 33.7 59.9 35.3 LY255,582 6.4 1.9 14.5 11.9 35.1 86.4 0.0 9 > 145 62.9 0.6 8.8 11.6 72.7 6.8 10 > 145 7.6 0.0 9.5 27.7 66.4 0.0 11 110.2 13.6 12.1 4.8 9.1 45.4 0.0 12 > 145 41.9 1.0 13.1 1.6 61.2 7.8 14 > 145 8.3 0.0 4.9 40.8 46.4 0.015 > 145 > 145 0.0 9.7 21.7 35.2 3.8Among the clinical opioid antagonists evaluated, diprenorphine (DPN) lacked metabolic stability (ti / 2 = 4.8 min), but NLX, NTX, and NLM had good metabolic stability (ti / 2 > 145 min) in human liver microsomes. Compounds 9, 10, 12, 14, and 15 also possessed good metabolic stability (ti / 2 > 145 min) in human liver microsomes. The results in mouse liver microsomes were different. Among the clinical opioid antagonists evaluated, NLM had the best metabolic stability (ti / 2 = 87.1 min), followed by NTX (ti / 2 = 57.4 min) and NLX (ti / 2 = 14.4 min). Compound 15 had the best metabolic stability (ti / 2 > 145 min), followed by Compound 9 (ti / 2 = 62.9 min). Compound 12 (ti / 2 = 41.9 min) and Compound 11 (ti / 2 = 13.6 min). Compounds 10 (ti / 2 = 7.6 min) and 14 (ti / 2 = 8.3 min) were found to have poor metabolic stability (defined as less than NLX).

[0148] Having identified several antagonists with a half-life of greater than 145 minutes in human liver microsomes, the ability of these analogues to inhibit major human CYP enzymes at 10 pM were profiled. CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 were selected for evaluation because they are related to more than 90% of drug4310712877:vlmetabolism and drug-drug interactions on the market and play an important role in the activation of a variety of endogenous and exogenous substances. While most of the analogues were found to inhibit CYP2D6 to a greater extent than NTX, most analogues were in the general range of inhibition seen with NLM.General Procedure for Vocal Cord Closure.

[0149] Male and female Sprague-Dawley rats (n=4; 28-29 weeks old) were single housed with free access to food and water in the home cage. Rats were maintained on a 12 hr light: dark cycle.

[0150] Five days prior to vocal cord recordings, rats were implanted with chronically indwelling i.v. catheters under ketamine (Schein, Dublin, OH, USA) / xylazine (Akom, Inc., Decatur, IL, USA) / acepromazine (Boehringer Ingelheim, St. Joseph, MO, USA; 75, 7.5, 0.75 mg / kg; i.p.) anesthesia. Following surgery recovery, rats w ere transported to a procedure room and glycopyrrolate (250 pg / kg; i.p.) was administered 30 min prior to the start of testing. Rats were then anesthetized with isoflurane and transferred to a custom board to maintain a 45° angle with the head up. Once positioned, an ear wax removal device with LED lights that incorporated a camera equipped with a spatula extension 0.7 cm from the tip was used to depress the velum, allowing for visualization of the vocal cords until the end of testing. Following a baseline 30-sec recording, rats were administered fentanyl (20 pg / kg; i.v.), followed 10 sec later by Compound 9 (1 mg / kg; i.v.), NLX (1 mg / kg; i.v.) or vehicle (10% captisol, i.v.) and vocal cords were recorded for 60 sec. Vocal cord closure was measured as a dichotomous qualitative variable, in which the vocal cords w ere determined to be either completely closed or completely open. Rescue agents were given in counterbalanced order across rats. If vocal cord opening failed to resume within 60 sec after treatment with Compound 9 or captisol, NLX (1 mg / kg; i.v.) was administered as a rescue.Example 8. Vocal Cord Closure Testing.

[0151] Intravenous fentanyl (10 mg / kg) produced a rapid and sustained vocal cord closure in rats, able to reversed by both Compound 9 and NLX, but not the vehicle (FIG. 10). There was no significant difference in vocal cord closure duration between Compound 9 and NLX treatments, but for both Compound 9 and NLX there was a significant difference from the vehicle rescue.General Procedure for Acute Withdrawal Testing.

[0152] A separate group of rats (n=4, 2 males and 2 females) were used to assess whether Compound 9 produced any opioid-induced withdrawal signs when given alone or4410712877:vlfollowing acute carfentanil. Each rat was injected with carfentanil (15 pg / kg, s.c.) or vehicle, followed 15 min later by Compound 9 (1.7 mg / kg, s.c.) or vehicle. Immediately after the second injection, rats were placed into a rectangular clear Plexiglas chamber with sani-chip bedding for 30 min. Each rat was assessed under each drug treatment condition, with the order of treatments being counterbalanced across rats and a minimum of 2 days between each test session. Videotaped records were assessed in by 3 independent “blind” observers using a time sampling of behaviors from 0-5, 10-15 and 20-25 min across the test duration. Based on previous work, the following behaviors were identified: (1) number of wet dog shakes; (2) number of digs; (3) number of grooms / licks; (4) number of vertical rears; (5) number of horizontal line crosses; (6) incidence of piloerection (yes / no); and (7) incidence of ptosis (yes / no).General Procedures for Data Analysis.

[0153] Antinociception data was measured as paw lick latency and used to calculate maximal percent of efficacy (%MPE) as seen in Equation (1):(paw lick latency— baseline measurement's - ~.- -Z30-base -l -ine measurement / x 100 Equation (11)

[0154] If an animal’s baseline value was over 10 sec, they were excluded from the analysis to account for tolerance to the hotplate. If an animal received a value under 25 sec without the antagonist being administered, they were also removed from the analysis to account for tolerance to fentanyl. GraphPad version 10.2.3 was used to generate a dose-response log(inhibitor) variable slope non-linear regression.

[0155] Locomotion and WBP data from the first 5 min was considered habituation and not included in the analysis. Locomotion was reported as mean (± SEM) distance traveled in cm and respiration was reported as mean (± SEM) minute ventilation in mL / min / g. Minute ventilation was used as the primary outcome measure as it is derived from breathing rate and tidal volume and is most closely associated with blood CO2 levels. For each dependent variable, mixed-effects analyses of variance (ANOVAs) were conducted using GraphPad Prism version 8.2.0, with subsequent post hoc comparisons made using a paired t-test, Dunnett’s, Tukey’s, or Sidak’s multiple comparisons test. Since previous work indicates that there are no reliable sex differences in fentanyl-induced respiratory depression, all primary analyses were collapsed across males and females.

[0156] For the behavioral checklist data, because there were no targeted behavioral signs observed under some treatment conditions, a non-parametric Mann-Whitney U test was used to assess behaviors across the 5-min observation periods.4510712877:vl

[0157] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0158] It will be understood that various details of the presently disclosed subject matter can be changed without departing from the scope of the subject matter disclosed herein. Furthermore, the foregoing description is for the purpose of illustration only, and not for the purpose of limitation.4610712877:vl

Claims

CLAIMSWhat is claimed is:

1. A compound represented by one of Formula I' « * — L — » (RA! B' ’(Rb)bLb— B Formula B’Formula Tor a co-crystal, hydrate, solvate, pharmaceutically acceptable salt, or combination thereof, wherein:A and C are rings;B is Ci-8 alkyl optionally substituted with at least one Rb, or is represented by a ring of Formula B’;rings A, B’, and C are each independently i) a first ring, ii) a second ring, iii) a first ring fused to another first ring, iv) a second ring fused to another second ring, or (v) a first ring fused to a second ring, wherein the first ring comprises a 5-membered carbocyclic ring, or a 5- membered heterocyclic ring, and the second ring comprises a 6-membered carbocyclic ring, or a 6-membered heterocyclic ring;a, b, and c are each independently 0 to 5, wherein the sum of a, b, and c is at least 1, Ra, Rb, and Rc are each independently Z, a C6-C30 ether, halogen, deuterium, OH, CN, Cj- C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, C3-C6 cycloalkyl, Ci-Ce alkyl(Cs-C6 cycloalkyl), Ci- Ce alkyl(Ca-C6 cycloalkenyl), C3-C6 cycloalkenyl, C2-C6 heterocycloalkyl, C2- Cb heterocycloalkenyl, Ci-Ce alkyl(C2-C6 heterocycloalkyl), Cj-Cf, alkyl(C2- Ce heterocycloalkenyl), C5-C12 aryl, C2-C12 heteroaryl, Ci-Ce alkyl (C2-C12 heteroaryl), each optionally substituted with deuterium, halogen, OH, CN, C1-C3 alkyl, C1-C3 haloalkyl, or a combination thereof, wherein any methylene may be replaced with O, S, Si, Ge, P, C(=O), Si ()). or NRi0;each occurrence of R10is independently hydrogen, deuterium, Ci-Cs alkyl in which any carbon-carbon single bond is optionally replaced by a carbon-carbon double or triple bond, and the C1-C6 alkyl is optionally substituted with deuterium, halogen, hydroxyl, cyano, or a combination thereof;4710712877:vlprovided that (i) the compound of Formula I comprises a C6-C30 ether; or (ii) the compound of Formula I comprises at least one Z; (iii) or the compound of Formula I comprises a C6-C30 ether and at least one Z;Z is Z’ or Yi-Z’, wherein: Z’ is hydroxyl (-OH ), thiol (-SH), cyano (-CN), fluorine (-F), isonitrile (-NC), nitro (-NO2), -X O(Y:). NH2, -NH-Y2, -N(Y2)2, -O-Y2, -S-Y2, -S(=O)(Y2), -S( ())( Y2)(N Y;). -S(=O)2(Y2), aldehyde ( (!!( <))). -C-; O: Y;. -('( S )Y;. ■■()•( t ()|Y2. -O- Ci S)Y2. -S-C(=O)Y2, -S-C(=S)Y2, -C(=O)-OY2, -C(=S)Y2, -C(=S)-OY2, -C(=O)-SY2, -C(=S)- SY2, -O-C(= Oj-OY2. -O-CI S)-OY2. -()-('( 0}-SY2. -O-C(===S)-SY2, -S-C(= ())-OY2. -S-( ( S}- OY2, -S-C(=O> SY2, -S-C(= =S)-SY2, -CI =O)-NH2, -C(=O)-NH(Y2), -C( =O)-N( Y2)2, -C(=S)-NH2, -( ( S)-Xi 1( Y;). -('( S)-X(Y2-NI!-('( ())-Y2. -N( Y2)-( ( O)-Y;. -Xi L('( S)-Y2. -N(Y2Ci S:-Y;. -XI l-S( Ok-Y2. -\( Y.')-S( O).-Y;. -()-( ( O)-X11.-. -0-C(==O)-NH(Y2), -()-('•: ()}- N( Y2)2, -O-C(= S ) -X 1 h, -O-C(==S)-NH(Y2), -O-C( S)-X( Yfo. -XH-Ct 0 }-Xl L. -XH-( ( ())■ NH(Y2), -NI l-C( ()L\( Y2)2, -X H -( ( S )- X M2-XI!-('( S)-\Ht Y2), -O-C( S)- N(Y2)2, - Xi b. Y2.}-( 1 O!- H2. -NH(Y2)-C(-O)-NH(Y2), -XI If Y2, I-( f O)-X( Y2b. -NH( Y2>-('( S )-NH2, - NH(Y2)-C(=S)-NH(Y2), -O-C(===S)~N(Y2)2, carboxylic acid (C(=O)OH), sulfonic acid (-SO3H), -C(=O)NH-NH2, -C(=S)NH-NH2, -S(=O)NH-NH2, -C(=O)NH-NH2, -C(=S)NH-NH2, or - C(::::NH-NH2)Y2; each occurrence of Yi and each occurrence of Y2independently comprise Cj-C12 alkyl, C2-C12 alkenyl, C2-Ci2alkynyl, C3-C6 cycloalkyl, Ci-Ce alkyl(C’3-C6 cycloalkyl), Ci-C6 alkyl(C3-C6 cycloalkenyl), C3-C6 cycloalkenyl, C2-C6 heterocycloalkyl, C2-Cfi heterocycloalkenyl, Ci-Cs alkyl(C2-C6 heterocycloalkyl), Ci-Cs alky i(C'~C.5 heterocycloalkenyl), C5-C12 aryl, C2-Ci2heteroaryl, Ci-C& alkyl (C2-C12 heteroaryl), each optionally substituted with halogen, deuterium, or a combination thereof;Laand Lb are each a single bond or Ci-Cs alkyl in which any carbon-carbon single bond is optionally replaced by a carbon-carbon double or triple bond, any methylene is optionally replaced by O, S, Si, Ge, P, C(=O), S( O j. or NR10, and Laand Lb are each optionally substituted with Z, deuterium, halogen, OH, CN, C1-C3 alkyl, C1-C3 haloalkyl, or a combination thereof, provided that Laand Lb are not simultaneously a single bond.

2. The compound of claim 1, wherein the compound of Formula I comprises a Ce- C30 ether.4810712877:vl3. The compound of claim 1, wherein the C6-C30 ether comprises the formula LG-polyether, wherein LG is a linking group comprising an ester, an amide, a urea, a carbamate, a carbonate, a thioester, a thioamide, a thiourea, a thiocarbonate, or a thiocarbamate.

4. The compound of claim 1, wherein the compound of Formula I comprises at least one Z.

5. The compound of claim 1, wherein the compound of Formula I comprises a Ce-C30 ether, and at least one Z.

6. The compound of claim 1, wherein ring A or ring B is a fused ring, and the compound of Formula I comprises a C6-C30 polyether.

7. The compound of claim 1, wherein ring A or ring B is a fused ring, and wherein the compound of Formula I comprises at least one Z.

8. The compound of claim 1, wherein Laand Lb are not single bonds.

9. The compound of claim 1, wherein Laand Lb are not single bonds, and at least one of ring A and ring B is a fused ring.

10. The compound of claim 1, wherein at least one of ring A and ring B are each independently a first ring fused to a second ring.

11. The compound of claim 1, wherein Laand Lb are not single bonds, and at least one of ring A and ring B is a first ring fused to a second ring.

12. The compound of claim 1, wherein the C6-C30 ether is a polyether.

13. The compound of claim 1, wherein Laand Lb are not single bonds, and the C6-C30 ether is a polyether.

14. The compound of claim 1, wherein one of Laand Lb is a C1-3 alkyl in which any methylene is optionally replaced by C(=O), O, S, or NR10, and the other of Laand Lb is a C3-64910712877:vlalkyl in which any methylene is optionally replaced by C(=O), O, S, or NR10, and any methylene is optionally substituted with deuterium, fluoro, or a combination thereof.

15. The compound of claim 1, wherein one of Laand Lb is O Laand Lb is (CH2)mN(R10)(CH2)n, wherein m and n are each 0 to 5, and the sum of m + n is at least 1, and any methylene is optionally substituted with deuterium, fluoro, or a combination thereof.

16. The compound of claim 1, wherein one of Laand Lb is O and the other of Laand Lb is (CH2)mN(R10)(CH2)n, wherein m and n are each 0 to 5, and the sum of m + n is at least 1, and any methylene is optionally substituted with deuterium, fluoro, or a combination thereof.

17. The compound of Claim 1, wherein the compound is represented by one of Formulas LA to I-GI-G wherein Ai-Ae are each independently C, CH, C(Ra), or N, Bi-EL are each independently C, CH, C(Rb), or N, and Ci-Ce are each independently C, CH, C(Rc), or N, and Xi-Xg are each independently N, O, S, C(=O), S(=O), S(=O)2, NH, or NR10.

18. The compound of Claim 1, wherein the compound is by Formula II5010712877:vlwherein Ci, C4, and C5 are each independently C, CH, C(Rc), or N, and E is a C6-C30 polyether.

19. The compound of Claim 18, whereinA is represented by one of the following formulaswherein Ai, A2, and A4- 5 are each independently C, CH, C(Ra), or N, and X1-X3 are each independently CH2, CHRX, C(RX)2, N, O, S, C(=O), S(=O), S(=O)2, NH, or NR10; and B’ is represented by one of the following formulaswherein Bi-Be are each independently C, CH, C(Rb), or N, and X4-X6 are each independently CH2, CHRX, C(RX)2, N, O, S, C(=O), S(=O), S(=O)2, NH, or NR10.

20. The compound of Claim 1, wherein the compound of Formula I is represented by one of Formulas III- A to III-D5110712877:vlFormula III-A Formula III-BFormula III-C Formula III-Dwherein Ai-Ae are each independently C, CH, C(Ra), or N; B1-B4 and Be are each independently C, CH, C(Rb), or N; X4-X6 are each independently CH2, CHRX, C(RX)2, N, O, S, C(=O), S(=O), S(=O)2, NH, or NR10; R10is hydrogen, deuterium or C1-3 alkyl optionally substituted with deuterium, halogen, or a combination thereof; n is 0-4; wherein any hydrogen of a methylene is optionally replaced with deuterium, or halogen.

21. The compound of Claim 17, wherein Z is present and is Z’.

22. The compound of Claim 1, wherein the compound is one of the following compounds5210712877:vl9: X = N, R = H, R' = CH2CH2Ph; 10: X = CH. R = H. R' = CH2CH2Ph: 11: X = N, R = Cl, R' = CH2CH, Ph; 12: X = N, R = OCH3, R' = CH2CH2Ph; 13: X = N, R = OCH, Ph, R' = CH2CH2Ph; 14: X = N, R = OCH,, R' = cyclohexylethyl 15: X = N, R = OCH3. R' = CH2CH2CH(CH3)2; 16: X = N. R = OCH3, R' = 2-indanylJWP-366JWP-231JWP-2305310712877:vlH2N^OJWP-271EW-710 EW-72523. A pharmaceutical composition comprising a compound of claim 1 and a pharmaceutically acceptable excipient.

24. A method for treating opioid overdose in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a compound of claim 1.5410712877:vl25. A method for treating a patient having an opioid use disorder according to a harm reduction protocol, comprising administering of a therapeutically effective amount of a compound of claim 1 to the patient having an opioid use disorder.5510712877:vl

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