Neuroactive steroids, compositions, and uses thereof

Abstract

Described herein are neuroactive steroids of the Formula (I): (see formula I), and pharmaceutically acceptable salts thereof. Such compounds are envisioned, in certain embodiments, to behave as GABA modulators, and which may be used for treatment, e.g., such for inducing sedation and / or anesthesia. The present invention also provides compounds of Formulae (A13) and (A16): (see formula A13) (see formula A16), and pharmaceutically acceptable salts thereof.

Description

92460299 NEUROACTIVE STEROIDS, COMPOSITIONS, AND USES THEREOF RELATED APPLICATIONS [1] The present application claims priority to patent applications, U.S.S.N. 61 / 869,440 filed August 23, 2013, U.S.S.N. 61 / 869,446 filed August 23, 2013, and U.S.S.N. 62 / 014,018 filed June 18, 2014. This is a divisional of Canadian patent application no. 2921512 filed on August 22, 2014. Background of the Invention [2] Brain excitability is defined as the level of arousal of an animal, a continuum that ranges from coma to convulsions, and is regulated by various neurotransmitters. In general, neurotransmitters are responsible for regulating the conductance of ions across neuronal membranes. At rest, the neuronal membrane possesses a potential ( or membrane voltage) of approximately-70 mV, the cell interior being negative with respect to the cell exterior. The potential (voltage) is the result of ion (K+, Na+, CC organic anions) balance across the neuronal semipenneable membrane. Neurotransmitters are stored in presynaptic vesicles and are released under the influence of neuronal action potentials. When released into the synaptic cleft, an excitatory chemical transmitter such as acetylcholine will cause membrane depolarization, e.g., a change of potential from-70 mV to -50 mV. This effect is mediated by postsynaptic nicotinic receptors which are stimulated by acetylcholine to increase membrane permeability to Na+ ions. The reduced membrane potential stimulates neuronal excitability in the form of a postsynaptic action potential. [3] In the case of the GABA receptor complex (GRC), the effect on brain excitability is mediated by GABA, a neurotransmitter. GABA has a profound influence on overall brain excitability because up to 40% of the neurons in the brain utilize GABA as a neurotransmitter. GABA regulates the excitability of individual neurons by regulating the conductance of chloride ions across the neuronal membrane. GABA interacts with its recognition site on the GRC to Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 facilitate the flow of chloride ions down an electrochemical gradient of the GRC into the cell. An intracellular increase in the levels of this anion causes hyperpolarization of the transmembrane potential, rendering the neuron less susceptible to excitatory inputs, i.e., reduced neuron excitability. In other words, the higher the chloride ion concentration in the neuron, the lower the brain excitability and level of arousal. [4] It is well-documented that the GRC is responsible for the mediation of anxiety, seizure activity, and sedation. Thus, GABA and drugs that act like GABA or facilitate the effects of GABA (e.g., the therapeutically useful barbiturates and benzodiazepines (BZs), such as Valium'ID) produce their therapeutically useful effects by interacting with specific regulatory sites on the GRC. Accumulated evidence has now indicated that in addition to the benzodiazepine and barbiturate binding site, the GRC contains a distinct site for neuroactive steroids. See, e.g., Lan, N. C. et al .. Neurochem. Res. (1991) 16:347-356. [5] Neuroactive steroids can occur endogenously. The most potent endogenous neuroactive steroids are 3a-hydroxy-5-reduced pregnan-20-one and 3a-2 l-dihydroxy-5-reduced pregnan- 20-one, metabolites of honnonal steroids progesterone and deoxycorticosterone, respectively. The ability of these steroid metabolites to alter brain excitability was recognized in 1986 (Majewska, M. D. et al., Science 232:1004-1007 (1986); Harrison, N. L. et al., J Pharmaco / . Exp. 1'her. 241:346-353 (1987)). [6] The ovarian honnone progesterone and its metabolites have been demonstrated to have profound effects on brain excitability (Backstrom, T. et al., Acta Obstet. Gynecol. Scand. Suppl. 130:19-24 (1985); Pfaff, D.W and McEwen, B. S., Science 219:808-814 (1983); Gyermek et al., J Med Chem. 11: 117 (1968); Lambert, J. et al., Trends Pharmacol. Sci. 8:224-227 (1987)). The levels of progesterone and its metabolites vary with the phases of the menstmal cycle. It has been well documented that the levels of progesterone and its metabolites decrease prior to the onset of menses. The monthly recurrence of certain physical symptoms prior to the onset of menses has also been well documented. These symptoms, which have become associated with premenstmal syndrome (PMS), include stress, anxiety, and migraine headaches (Dalton, K., Premenstrual Syndrome and Progesterone Therapy, 2nd edition, Chicago Yearbook, Chicago (1984)). Subjects with PMS have a monthly recurrence of symptoms that are present in premenses and absent in postmenses. 2 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 [7] In a similar fashion, a reduction in progesterone has also been temporally correlated with an increase in seizure frequency in female epileptics, i.e., catamenial epilepsy (Laidlaw, J., Lancet, 1235-1237 (1956)). A more direct correlation has been observed with a reduction in progesterone metabolites (Roscisze\vska et al., J. Neural. Neurosurg. Psych. 49:4 7-51 ( 1986)). In addition, for subjects with primary generalized petit mal epilepsy, the temporal incidence of seizures has been correlated with the incidence of the symptoms of premenstmal syndrome (Backstrom, T. et al., J Psychosom. Obstet. Gynaecol. 2:8-20 (1983)). The steroid deoxycorticosterone has been found to be effective in treating subjects with epileptic spells correlated with their menstmal cycles (Aird, R.B. and Gordan, G., J. Amer. Med. Soc. 145:715- 719 (1951)). [8] A syndrome also related to low progesterone levels is postnatal depression (PND). Immediately after birth, progesterone levels decrease dramatically leading to the onset of PND. The symptoms of PND range from mild depression to psychosis requiring hospitalization. PND is also associated with severe anxiety and irritability. PND-associated depression is not amenable to treatment by classic antidepressants, and women experiencing PND show an increased incidence of PMS (Dalton, K., Premenstrual Syndrome and Progesterone Therapy, 2nd edition, Chicago Yearbook, Chicago (1984)). [9] Collectively, these observations imply a cmcial role for progesterone and deoxycorticosterone and more specifically their metabolites in the homeostatic regulation of brain excitability, which is manifested as an increase in seizure activity or symptoms associated with catamenial epilepsy, PMS, and PND. The correlation behveen reduced levels of progesterone and the symptoms associated with PMS, PND, and catamenial epilepsy (Backstrom, T. et al., J Psychosom.Obstet. Gynaecol. 2:8-20 (1983 )); Dalton, K., Premenstrnal Syndrome and Progesterone Therapy, 2nd edition, Chicago Yearbook, Chicago (1984)) has prompted the use of progesterone in their treatment (Mattson et al., "Medroxyprogesterone therapy of catamenial epilepsy," in Advances in Epileptology: XVth Epilepsy International Symposium, Raven Press, New York (1984), pp. 279-282, and Dalton, K., Premenstrual Syndrome and Progesterone Therapy, 2nd edition, Chicago Yearbook, Chicago (1984)). However, progesterone is not consistently effective in the treatment of the aforementioned syndromes. For example, no dose-response relationship exists for progesterone in the treatment 3 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 of PMS (Maddocks et al., Obstet. Gynecol. 154:573-581 (1986); Dennerstein et al., Brit. Med J 290:16-17 (1986)). [10) New and improved neuroactive steroids are needed that act as modulating agents for brain excitability, as well as agents for the prevention and treatment of CNS-related diseases. The compounds, compositions, and methods described herein are directed toward this end. Summary of the Invention

[11] Provided herein are C2 l-substituted neuroactive steroids designed, for example, to act as GABA modulators. In certain embodiments, such compounds are envisioned to be useful as therapeutic agents for the inducement of anesthesia and / or sedation in a subject. In some embodiments, such compounds are envisioned to be useful as therapeutic agents for treating a CNS-related disorder (e.g., sleep disorder, a mood disorder, a schizophrenia spectrum disorder, a convulsive disorder, a disorder of memory and / or cognition, a movement disorder, a personality disorder, autism spectmm disorder, pain, traumatic brain injury, a vascular disease, a substance abuse disorder and / or withdrawal syndrome, or tinnitus) in a subject in need (e.g., a subject with Rett syndrome, Fragile X syndrome, or Angelman syndrome). [12) In one aspect, provided is a compound ofFonnula (I): HO''' R2 0 (;) L: (I), a pharmaceutically acceptable salt thereof, wherein: A is an optionally substituted nitrogencontaining heteroaryl or heterocyclyl; L is-C(R3)(R~)-, -0-, -S-, or-NR3 -; R1 is hydrogen or C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, carbocyclyl, or heterocyclyl; R2 is hydrogen, C1-C6 alkyl (e.g., C1-C6 haloalkyl), or C1-C6 alkoxy; each R3 is independently hydrogen or C1-C6 alkyl; 4 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 R5 is absent or hydrogen; and ::..:.:.:.::..: represents a single or double bond, wherein when one of ::..:.:.:.::..: is a double bond, the other ::..:.:.:.::..: is a single bond; and when one of the ::..:.:.:.::..: is a double bond, R5 is absent.

[13] The present invention also provides pharmaceutical compositions comprising a compound of the present invention and methods of use and treatment, e.g., such as for inducing sedation and / or anesthesia, for treating a CNS-related disorder.

[14] Steroids of Formula (I), sub-genera thereof, and pham1aceutically acceptable salts thereof are collectively referred to herein as "compounds of the present invention."

[15] In another aspect, provided is a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable excipient. In certain embodiments, the compound of the present invention is provided in an effective amount in the pharmaceutical composition. In certain embodiments, the compound of the present invention is provided in a therapeutically effective amount. In certain embodiments, the compound of the present invention is provided in a prophylactically effective amount.

[16] Compounds of the present invention as described herein, act, in certain embodiments, as GABA modulators, e.g., effecting the GABA.A. receptor in either a positive or negative manner. As modulators of the excitability of the central nervous system (CNS), as mediated by their ability to modulate GABA.A receptor, such compounds are expected to have CNS-activity. [ 17] Thus, in another aspect, provided are methods of treating a CNS-related disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound of the present invention. In certain embodiments, the CNS-related disorder is selected from the group consisting of a sleep disorder, a mood disorder, a schizophrenia spectmm disorder, a convulsive disorder, a disorder of memory and / or cognition, a movement disorder, a personality disorder, autism spectmm disorder, pain, traumatic brain injury, a vascular disease, a substance abuse disorder and / or withdrawal syndrome, and tinnitus. In certain embodiments, the compound is administered orally, subcutaneously, intravenously, or intramuscularly. In certain embodiments, the compound is administered chronically. In certain embodiments, the compound is administered continuously, e.g., by continuous intravenous infusion. 5 Date Rec;ue / Date Received 2024-04-12 84019316

[18] Other objects and advantages will become apparent to those skilled in the art from a consideration of the ensuing Detailed Description, and Examples. Definitions Chemical definitions

[19] Definitions of specific functional groups and chemical terms are described in more detail below. The chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Thomas Sorrell, Organic Chemistry, University Science Books, Sausalito, 1999; Smith and March, March's Advanced Organic Chemistry, 5th Edition, John Wiley & Sons, Inc., New York, 200 I; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; and Carruthers, Some Modern Methods a / Organic Synthesis, 3rd Edition, Cambridge University Press, Cambridge, 1987.

[20] Compounds described herein can comprise one or more asymmetric centers, and thus can exist in various isomeric fom1s, e.g., enantiomers and / or diastereomers. For example, the compounds described herein can be in the form of an individual enantiomer, diastereomer or geometric isomer, or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer. Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric syntheses. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981 ); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistry a / Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions p. 268 (E.L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972). The invention additionally encompasses compounds described herein as individual isomers substantially free of other isomers, and alternatively, as mixtures of various isomers. 6 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[21] As used herein a pure enantiomeric compound is substantially free from other enantiomers or stereoisomers of the compound (i.e., in enantiomeric excess). In other words, an "S" form of the compound is substantially free from the "R" fom1 of the compound and is, thus, in enantiomeric excess of the "R" form. The term "enantiomerically pure" or "pure enantiomer" denotes that the compound comprises more than 75% by weight, more than 80% by weight, more than 85% by weight, more than 90% by weight, more than 91 % by weight, more than 92% by weight, more than 93% by weight, more than 94% by weight, more than 95% by weight, more than 96% by weight, more than 97% by weight, more than 98% by weight, more than 98.5% by weight, more than 99% by weight, more than 99.2% by weight, more than 99.5% by weight, more than 99.6% by weight, more than 99.7% by weight, more than 99.8% by weight or more than 99 .9% by weight, of the enantiomer. In certain embodiments, the weights are based upon total weight of all enantiomers or stereoisomers of the compound.

[22] In the compositions provided herein, an enantiomerically pure compound can be present with other active or inactive ingredients. For example, a pharmaceutical composition comprising enantiomerically pure R-compound can comprise, for example, about 90% excipient and about 10% enantiomerically pure R-compound. In certain embodiments, the enantiomerically pure R-compound in such compositions can, for example, comprise, at least about 95% by weight R-compound and at most about 5% by weight S-compound, by total weight of the compound. For example, a pharmaceutical composition comprising enantiomerically pure S-compound can comprise, for example, about 90% excipient and about 10% enantiomerically pure S-compound. In certain embodiments, the enantiomerically pure Scompound in such compositions can, for example, comprise, at least about 95% by weight Scompound and at most about 5% by weight R-compound, by total weight of the compound. In certain embodiments, the active ingredient can be formulated with little or no excipient or earner.

[23] Compound described herein may also comprise one or more isotopic substitutions. For example, H may be in any isotopic form, including 1H, 2H (D or deuterium), and 3H (T or tritium); C may be in any isotopic form, including 12C, 13C, and 14C; 0 may be in any isotopic fom1, including 160 and 180; and the like. 7 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[24] The articles "a" and "an" may be used herein to refer to one or to more than one (i.e. at least one) of the grammatical objects of the article. By way of example "an analogue" means one analogue or more than one analogue. (25] When a range of values is listed, it is intended to encompass each value and sub-range within the range. For example "C1---6 alkyl" is intended to encompass, C1, C2, C3, C4, Cs, C6, C1- 6, C1-s, C1--4, C1-3, C1-2, C2-6, C2-s, C2-4, C2-3, C3---6, C3_5, C3--4, C4-6, C4-s, and Cs---{i alkyl.

[26] The following terms are intended to have the meanings presented therewith below and are useful in understanding the description and intended scope of the present invention. (27] "Alkyl" refers to a radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 20 carbon atoms ("C1_20 alkyl"). In some embodiments, an alkyl group has 1 to 12 carbon atoms ("C1_12 alkyl"). In some embodiments, an alkyl group has I to 8 carbon atoms ("C1-s alkyl"). In some embodiments, an alkyl group has 1 to 6 carbon atoms ("C1-6 alkyl", also referred to herein as "lower alkyl"). In some embodiments, an alkyl group has 1 to 5 carbon atoms ("C1-s alkyl"). In some embodiments, an alkyl group has 1 to 4 carbon atoms ("C1--4 alkyl"). In some embodiments, an alkyl group has 1 to 3 carbon atoms ("C1_3 alkyl"). In some embodiments, an alkyl group has I to 2 carbon atoms ("C1_2 alkyl"). In some embodiments, an alkyl group has I carbon atom ("C1 alkyl"). In some embodiments, an alkyl group has 2 to 6 carbon atoms ("C2---6 alkyl"). Examples of C l---6 alkyl groups include methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), iso-butyl (C4), n-pentyl (Cs), 3-pentanyl (C5), amyl (C5), neopentyl (C5), 3-methyl-2-butanyl (C5), tertiary amyl (Cs), and n-hexyl (C6)- Additional examples of alkyl groups include n-heptyl (C1), n-octyl (Cs) and the like. Unless otherwise specified, each instance of an alkyl group is independently optionally substituted, i.e., tmsubstituted (an "unsubstituted alkyl") or substituted (a "substituted alkyl") vvith one or more substituents; e.g., for instance from 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In certain embodiments, the alkyl group is unsubstituted C1-10 alkyl (e.g., -CH3). In certain embodiments, the alkyl group is substituted C1-10 alkyl. Common alkyl abbreviations include Me (-CH3), Et (-CH2CH3), iPr (-CH(CH3)2), nPr (CH2CH2CH3), n-Bu (-CH2CH2CH2CH3), or i-Bu (-CH2CH(CH3)i).

[28] "Alkenyl" refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 20 carbon atoms, one or more carbon-carbon double bonds, and no triple bonds ("C2- 8 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 20 alkenyl"). In some embodiments, an alkenyl group has 2 to 10 carbon atoms ("C2-io alkenyl"). In some embodiments, an alkenyl group has 2 to 8 carbon atoms ("C28 alkenyl"). In some embodiments, an alkenyl group has 2 to 6 carbon atoms ("C2---6 alkenyl"). In some embodiments, an alkenyl group has 2 to 5 carbon atoms ("C2_5 alkenyl"). In some embodiments, an alkenyl group has 2 to 4 carbon atoms ("C2-4 alkenyl"). In some embodiments, an alkenyl group has 2 to 3 carbon atoms ("C2-3 alkenyl"). In some embodiments, an alkenyl group has 2 carbon atoms ("C2 alkenyl"). The one or more carboncarbon double bonds can be internal (such as in 2-butenyl) or tenninal (such as in 1-butenyl). Examples of C2-4 alkenyl groups include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1- butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like. Examples of C2-6 alkenyl groups include the aforementioned C2-4 alkenyl groups as well as pentenyl (C5), pentadienyl (C5), hexenyl (C6), and the like. Additional examples of alkenyl include heptenyl (C7), octenyl (C8), octatrienyl (Cg), and the like. Unless otherwise specified, each instance of an alkenyl group is independently optionally substituted, i.e., unsubstituted (an "unsubstituted alkenyl") or substituted (a "substituted alkenyl") with one or more substituents e.g., for instance from l to 5 substituents, 1 to 3 substituents, or 1 substituent. In certain embodiments, the alkenyl group is unsubstituted C2-10 alkenyl. In certain embodiments, the alkenyl group is substituted C2-10 alkenyl.

[29] "Alkynyl" refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 20 carbon atoms, one or more carbon-carbon triple bonds, and optionally one or more double bonds ("C2-20 alkynyl"). In some embodiments, an alkynyl group has 2 to 10 carbon atoms ("C2-10 alkynyl"). In some embodiments, an alkynyl group has 2 to 8 carbon atoms ("C2-s alkynyl"). In some embodiments, an alkynyl group has 2 to 6 carbon atoms ("C2_6 alkynyl"). In some embodiments, an alkynyl group has 2 to 5 carbon atoms ("C2_5 alkynyl"). In some embodiments, an alkynyl group has 2 to 4 carbon atoms ("C2_4 alkynyl"). In some embodiments, an alkynyl group has 2 to 3 carbon atoms ("C2-3 alkynyl"). In some embodiments, an alkynyl group has 2 carbon atoms ("C2 alkynyl"). The one or more carboncarbon triple bonds can be internal (such as in 2-butynyl) or terminal (such as in 1-butynyl). Examples of C2-4 alkynyl groups include, without limitation, ethynyl (C2), 1-propynyl (C.~), 2- propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), and the like. Examples of C2---6 alkenyl groups 9 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 include the aforementioned C2--4 alkynyl groups as well as pentynyl (Cs), hexynyl (CG), and the like. Additional examples of alkynyl include heptynyl (C7), octynyl (C&), and the like. Unless otherwise specified, each instance of an alkynyl group is independently optionally substituted, i.e., unsubstituted (an "unsubstituted alkynyl") or substituted (a "substituted alkynyl") with one or more substituents; e.g., for instance from I to 5 substituents, 1 to 3 substituents, or 1 substituent. In certain embodiments, the alkynyl group is unsubstituted C2-rn alkynyl. In certain embodiments, the alkynyl group is substituted C2_10 alkynyl. [30) "Aryl" refers to a radical of a monocyclic or polycyclic ( e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 re electrons shared in a cyclic array) having 6-14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system ("C6-i4 aryl"). In some embodiments, an aryl group has six ring carbon atoms ("CG aryl"; e.g., phenyl). In some embodiments, an aryl group has ten ring carbon atoms ("C 10 aryl"; e.g., naphthyl such as 1- naphthyl and 2-naphthyl). In some embodiments, an aryl group has fourteen ring carbon atoms ("C14 aryl"; e.g., anthracyl). "Aryl" also includes ring systems wherein the aryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the radical or point of attachment is on the aryl ring, and in such instances, the number of carbon atoms continue to designate the number of carbon atoms in the aryl ring system. Aryl groups include, but are not limited to, phenyl, naphthyl, indenyl, and tetrahydronaphthyl. Unless otherwise specified, each instance of an aryl group is independently optionally substituted, i.e., unsubstituted (an "unsubstituted aryl") or substituted (a "substituted aryl") with one or more substituents. In certain embodiments, the aryl group is unsubstituted CG-14 aryl. In certain embodiments, the aryl group is substituted C6_14 aryl. [31) In certain embodiments, an aryl group substituted with one or more of groups selected from halo, C1-Cs alkyl, C1-C8 haloalkyl, cyano, hydroxy, C1-Cs alkoxy, and amino. [32) Examples of representative substituted aryls include the following and wherein one ofR56 and R57 may be hydrogen and at least one ofR56 and R57 is each independently selected from C 1-Cs alkyl, C1-Cs haloalkyl, 4-10 membered heterocyclyl, Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 alkanoyl, C1-Cs alkoxy, heteroaryloxy, alkylamino, arylamino, heteroarylamino, NR58COR59, NR58SOR59 NR58SO2R59 , COOalkyl, COOaryl, CONR58R59, CONR58OR59 , NR58R59 , SO2NR58R59 , S-alkyl, SOalkyl, SO2alkyl, Saryl, SOaryl, SO2aryl; or R56 and R57 may be joined to form a cyclic ring (saturated or unsaturated) from 5 to 8 atoms, optionally containing one or more heteroatoms selected from the group N, 0, or S. R60 and R61 are independently hydrogen, C 1-Cs alkyl, C 1-C4 haloalkyl, C3-C10 cycloalkyl, 4-1 O membered heterocyclyl, C6-C10 aryl, substituted C6-C10 aryl, 5-10 membered heteroaryl, or substituted 5-10 membered heteroaryl .

[33] Other representative aryl groups having a fused heterocyclyl group include the following: wherein each Wis selected from C(R66)2, NR66, 0, and S; and each Y is selected from carbonyl, NR66, 0 and S; and R66 is independently hydrogen, Ci-Cs alkyl, C3-C10 cycloalkyl, 4-10 membered heterocyclyl, C6-C10 aryl, and 5-10 membered heteroaryl.

[34] "Halo" or "halogen," independently or as part of another substituent, mean, unless otherwise stated, a fluorine (F), chlorine (Cl), bromine (Br), or iodine (I) atom. The term "halide" by itself or as part of another substituent, refers to a fluoride, chloride, bromide, or iodide atom. In certain embodiments, the halo group is either fluorine or chlorine.

[35] "Haloalkyl" and "haloalkoxy" can include alkyl and alkoxy structures that are substituted with one or more halo groups or with combinations thereof. For example, the tem1s "fluoroalkyl" and "fluoroalkoxy" include haloalkyl and haloalkoxy groups, respectively, in which the halo is fluorine.

[36] "Heteroaryl" refers to a radical of a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 n electrons shared in a cyclic array) having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur (''5-10 membered heteroaryl"). In heteroaryl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. Heteroaryl bicyclic ring 11 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 systems can include one or more heteroatoms in one or both rings. "Heteroaryl" includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the point of attachment is on the heteroaryl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heteroaryl ring system. "Heteroaryl" also includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more aryl groups wherein the point of attachment is either on the aryl or heteroaryl ring, and in such instances, the number of ring members designates the number of ring members in the fused ( aryl / heteroaryl) ring system. Bicyclic heteroaryl groups wherein one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and the like) the point of attachment can be on either ring, i.e .. , either the ring bearing a heteroatom (e.g., 2-indolyl) or the ring that does not contain a heteroatom ( e.g., 5-indolyl). [37) In some embodiments, a heteroaryl group is a 5-10 membered aromatic ring system having ring carbon atoms and 1--4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (''5-10 membered heteroaryl"). In some embodiments, a heteroaryl group is a 5-8 membered aromatic ring system having ring carbon atoms and 1--4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-8 membered heteroaryl"). In some embodiments, a heteroaryl group is a 5-6 membered aromatic ring system having ring carbon atoms and 1--4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-6 membered heteroaryl"). In some embodiments, the 5-6 membered heteroaryl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, each instance of a heteroaryl group is independently optionally substituted, i.e., unsubstin1ted (an "unsubstituted heteroaryl") or substituted (a "substituted heteroaryl") with one or more substituents. In certain embodiments, the heteroaryl group is unsubstituted 5-14 membered heteroaryl. In certain embodiments, the heteroaryl group is substituted 5-14 membered heteroaryl. 12 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[38] Exemplary 5-membered heteroaryl groups containing one heteroatom include, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, without limitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, without limitation, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, without limitation, tetrazolyl. Exemplary 6~membered heteroaryl groups containing one heteroatom include, without limitation, pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, without limitation, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, without limitation, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, without limitation, azepinyl, oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groups include, without limitation, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, without limitation, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

[39] Examples of representative heteroaiyls include the following formulae: --0 ~'( / / \\ -F> ~~ (') () ..,N I( N y y y N.., N N 00 t N tN) ~ NJ N~ N N ~ V-✓ 13 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 wherein each Y is selected from carbonyl, N, NR65, 0, and S; and R65 is independently hydrogen, C1-Cs alkyl, C3-C10 cycloalkyl, 4-10 membered heterocyclyl, C6-C10 aryl, and 5-10 membered heteroaryl. [40) "Carbocyclyl" or "carbocyclic" refers to a radical of a non-aromatic cyclic hydrocarbon group having from 3 to 10 ring carbon atoms ("C3_10 carbocyclyl") and zero heteroatoms in the non-aromatic ring system. In some embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms ("C3_8 carbocyclyl"). In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms ("C.H carbocyclyl"). In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms ("C3-6 carbocyclyl"). In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms ("Cs-10 carbocyclyl"). Exemplary C3-6 carbocyclyl groups include, without limitation, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (Cs), cyclopentenyl (Cs), cyclohexyl (C5), cyclohexenyl (C6), cyclohexadienyl (C6), and the like. Exemplary C3_8 carbocyclyl groups include, without limitation, the aforementioned C3_6 carbocyclyl groups as well as cycloheptyl (C1), cycloheptenyl (C1), cycloheptadienyl (C1), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2.2.l]heptanyl (C7), bicyclo[2.2.2]octanyl (C8), and the like. Exemplary C3_10 carbocyclyl groups include, without limitation, the aforementioned C3-8 carbocyclyl groups as well as cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C10), cyclodecenyl (C10), octahydro-lH-indenyl (C9), decahydronaphthalenyl (C10), spiro[4.5]decanyl (C10), and the like. As the foregoing examples illustrate, in certain embodiments, the carbocyclyl group is either monocyclic ("monocyclic carbocyclyl") or contain a fused, bridged or spiro ring system such as a bicyclic system ("bicyclic carbocyclyl") and can be saturated or can be partially unsaturated. "Carbocyclyl" also includes ring systems wherein the carbocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups wherein the point of attachment is on the carbocyclyl ring, and in such instances, the number of carbons continue to designate the number of carbons in the carbocyclic ring system. Unless otherwise specified, each instance of a carbocyclyl group is independently optionally substituted, i.e., unsubstituted (an "unsubstituted carbocyclyl") or substituted (a "substituted carbocyclyl") with one or more substituents. In certain embodiments, the carbocyclyl group is unsubstituted C3-10 carbocyclyl. In certain embodiments, the carbocyclyl group is a substituted C3-10 carbocyclyl. 14 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[41] In some embodiments, "carbocyclyl" is a monocyclic, saturated carbocyclyl group having from 3 to IO ring carbon atoms ("C3 10 cycloalkyl"). In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms ("CH cycloalkyl"). In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms ("C3-u cycloalkyl"). In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms ("Cs-u cycloalkyl"). In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms ("Cs--10 cycloalkyl"). Examples of Cs--6 cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C5). Examples of C.H cycloalkyl groups include the aforementioned C5-u cycloalkyl groups as well as cyclopropyl (C3) and cyclobutyl (C4). Examples of C3-8 cycloalkyl groups include the aforementioned C3-u cycloalkyl groups as well as cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, each instance of a cycloalkyl group is independently unsubstituted (an "unsubstituted cycloalkyl") or substituted (a "substituted cycloalkyl") with one or more substituents. In certain embodiments, the cycloalkyl group is unsubstituted C,-10 cycloalkyl. In certain embodiments, the cycloalkyl group is substituted C,-10 cycloalkyl. [42) "Heterocyclyl" or "heterocyclic" refers to a radical of a 3- to l 0-membered nonaromatic ring system having ring carbon atoms and I to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon ("3-10 membered heterocyclyl"). In heterocyclyl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. A heterocyclyl group can either be monocyclic ("monocyclic heterocyclyl") or a fused, bridged or spiro ring system such as a bicyclic system ("bicyclic heterocyclyl"), and can be saturated or can be partially unsaturated. Heterocyclyl bicyclic ring systems can include one or more heteroatoms in one or both rings. "Heterocyclyl" also includes ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more carbocyclyl groups wherein the point of attachment is either on the carbocyclyl or heterocyclyl ring, or ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heterocyclyl ring system. Unless otherwise specified, each instance of heterocyclyl is independently optionally substituted, i.e., unsubstituted (an "unsubstituted heterocyclyl") or substituted (a "substituted 15 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 heterocyclyl") with one or more substituents. In certain embodiments, the heterocyclyl group is unsubstituted 3-10 membered heterocyclyl. In certain embodiments, the heterocyclyl group is substituted 3-10 membered heterocyclyl. (43] In some embodiments, a heterocyclyl group is a 5-10 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon ("5-10 membered heterocyclyl"). In some embodiments, a heterocyclyl group is a 5-8 membered nonaromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-8 membered heterocyclyl"). In some embodiments, a heterocyclyl group is a 5-6 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-6 membered heterocyclyl"). In some embodiments, the 5-6 membered heterocyclyl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocyclyl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocyclyl has one ring heteroatom selected from nitrogen, oxygen, and sulfur.

[44] Exemplary 3-membered heterocyclyl groups containing one heteroatom include, without limitation, azirdinyl, oxiranyl, thiorenyl. Exemplary 4-membered heterocyclyl groups containing one heteroatom include, without limitation, azetidinyl, oxetanyl and thietanyl. Exemplary 5-membered heterocyclyl groups containing one heteroatom include, without limitation, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groups containing two heteroatoms include, without limitation, dioxolanyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclyl groups containing three heteroatoms include, without limitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclyl groups containing one heteroatom include, without lin1itation, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6- membered heterocyclyl groups containing two heteroatoms include, without limitation, piperazinyl, morpholinyl, dithianyl, dioxanyl. Exemplary 6-membered heterocyclyl groups containing two heteroatoms include, without limitation, triazinanyl. Exemplary 7-membered 16 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 heterocyclyl groups containing one heteroatom include, without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary 8-membered heterocyclyl groups containing one heteroatom include, without limitation, azocanyl, oxecanyl and thiocanyl. Exemplary 5-membered heterocyclyl groups fused to a C6 aryl ring (also referred to herein as a 5,6-bicyclic heterocyclic ring) include, without limitation, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinonyl, and the like. Exemplary 6-membered heterocyclyl groups fused to an aryl ring (also referred to herein as a 6,6-bicyclic heterocyclic ring) include, without limitation, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like.

[45] Particular examples of heterocyclyl groups are shown in the following illustrative examples: wherein each Wis selected from CR67, C(R67)2, NR67, 0, and S; and each Y is selected from NR67, 0, and S; and R67 is independently hydrogen, C1-Cs alkyl, c.~-C10 cycloalkyl, 4-10 membered heterocyclyl, C6-C10 aryl, and 5-10-membered heteroaryl. These heterocyclyl rings may be optionally substituted with one or more groups selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl (e.g., amido), aminocarbonylamino, aminosulfonyl, sulfonylamino, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, halogen, hydroxy, keto, nitro, thiol, -S-alkyl, -Saryl, -S(O)-alkyl, -S(O)-aryl, -S(Oh-alkyl, and-S(O)2-aryl. Substituting groups include carbonyl or thiocarbonyl which provide, for example, lactam and urea derivatives.

[46] "Acyl" refers to a radical -C(O)R20 , where R20 is hydrogen, substituted or unsubstitued alkyl, substituted or unsubstitued alkenyl, substituted or m1substitued alkynyl, substituted or m1substitued carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted 17 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 aryl, or substituted or unsubstitued heteroaryl, as defined herein. "Alkanoyl" is an acyl group wherein R20 is a group other than hydrogen. Representative acyl groups include, but are not limited to, fom1yl (-CHO), acetyl (-C(=O)CH3), cyclohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl (-C(=O)Ph), benzylcarbonyl (-C(=O)CH2Ph), -C(O)-C1- Cs alkyl, -C(O)-{CH2)t(C6-C10 aryl}, -C(O)-(CH2)1(5-10 membered heteroaryl), -C(O}( CH2)1(C3-C10 cycloalkyl), and-C(O)-(CH2)1(4-10 membered heterocyclyl), wherein tis an integer from Oto 4. In certain embodiments, R21 is C1-C8 alkyl, substituted with halo or hydroxy; or CrC 10 cycloalkyl, 4-10 membered heterocyclyl, C6-C10 aryl, arylalkyl, 5-10 membered heteroaryl or heteroarylalkyl, each of which is substituted with unsubstituted C1-C4 alkyl, halo, unsubstituted C i-C4 alkoxy, unsubstituted C 1-C4 haloalkyl, unsubstituted C1-C4 hydroxyalkyl, or unsubstituted C1-C4 haloalkoxy or hydroxy. [47) "Acylamino" refers to a radical -NR22C(O)R23, where each instance ofR22 and R23 is independently hydrogen, substituted or unsubstitued alkyl, substituted or unsubstitued alkenyl, substituted or mm1bstitued alkynyl, substituted or unsubstitued carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or m1substitued heteroaryl,, as defined herein, or R22 is an amino protecting group. Exemplary "acylamino" groups include, but are not limited to, formylamino, acetylamino, cyclohexylcarbonylamino, cyclohexylmethyl-carbonylamino, benzoylamino and benzylcarbonylamino. Particular exemplaiy "acylamino" groups are -NR24C(O)-C1-C8 alkyl, -NR24C(O)-(CH2)1(C6-C10 aryl), - NR24C(O)-{CH2)t(5-10 membered heteroaiyl), -NR24C(O}-(CH2)1(C3-C10 cycloalkyl), andNR24C( O)-(CH2)t(4-10 membered heterocyclyl), wherein tis an integer from Oto 4, and each R24 independently represents hydrogen or Ci-Cs alkyl. In certain embodiments, R25 is H, Ci-Cs alkyl, substituted with halo or hydroxy; C3-C10 cycloalkyl, 4-10 membered heterocyclyl, C6- C 10 aryl, arylalkyl, 5-10 membered heteroaryl or heteroarylalkyl, each of which is substituted with unsubstituted C1-C4 alkyl, halo, unsubstituted C 1-C4 alkoxy, unsubstituted C1-C4 haloalkyl, unsubstituted C1-C4 hydroxyalkyl, or unsubstituted C1-C4 haloalkoxy or hydroxy; and R26 is H, C 1-C8 alkyl, substituted with halo or hydroxy; CrC 10 cycloalkyl, 4-10- membered heterocyclyl, C6-C 10 aryl, arylalkyl, 5-10-membered heteroaryl or heteroarylalkyl, each of which is substituted with unsubstituted C1-C4 alkyl, halo, m1substituted C1-C4 alkoxy, 18 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 unsubstituted C1-C4 haloalkyl, unsubstituted C 1-C4 hydroxyalkyl, or unsubstituted C 1-C4 haloalkoxy or hydroxy; provided at least one of R25 and R26 is other than H. [48) "Acyloxy" refers to a radical -OC(O)R27, where R27 is hydrogen, substituted or unsubstitued alkyl, substituted or unsubstitued alkenyl, substituted or unsubstitued alkynyl, substituted or unsubstitued carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstitued heteroaryl, as defmed herein. Representative examples include, but are not limited to, fom1yl, acetyl, cyclohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl, and benzylcarbonyl. In certain embodiments, R28 is C1-Cs alkyl, substituted with halo or hydroxy; C3-C 10 cycloalkyl, 4-10-membered heterocyclyl, C6- C10 aryl, arylalkyl, 5-10-membered heteroaryl or heteroarylalkyl, each of which is substituted with unsubstituted C1-C4 alkyl, halo, unsubstituted C 1-C4 alkoxy, unsubstituted C1-C4 haloalkyl, unsubstituted C1-C4 hydroxyalkyl, or unsubstituted C1-C4 haloalkoxy or hydroxy. [49) "Alkoxy" refers to the group -OR29 where R29 is substituted or unsubstituted alkyl, substituted or mm1bstitued alkenyl, substituted or unsubstitued alkynyl, substituted or unsubstitued carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstitued heteroaryl. Particular alkoxy groups are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, and 1,2- dimethylbutoxy. Particular alkoxy groups are lower alkoxy, i.e., with between 1 and 6 carbon atoms. Fmther particular alkoxy groups have between 1 and 4 carbon atoms. [50) In certain embodiments, R29 is a group that has 1 or more substituents, for instance from 1 to 5 substituents, and particularly from 1 to 3 substituents, in particular 1 substituent, selected from the group consisting of amino, substituted amino, C6-C10 aryl, aiyloxy, carboxyl, cyano, C3-C rn cycloalkyl, 4-10 membered heterocyclyl, halogen, 5-10 membered heteroaryl, hydroxy, nitro, thioalkoxy, thioai·yloxy, thiol, alkyl-S(O)-, aryl-S(O)-, alkyl-S(Oh- and aryl-S(O)2-. Exemplary "substituted alkoxy" groups include, but are not limited to, -O-(CH2)1(C6-C10 aryl), -O-(CH2)t(5-10 membered heteroaryl), -O--(CH2)r(C3-C10 cycloalkyl), and-O--(CH2)r(4-10 membered heterocyclyl), wherein t is an integer from O to 4 and any aryl, heteroaryl, cycloalkyl or heterocyclyl groups present, may themselves be substituted by unsubstituted C 1-C4 alkyl, halo, unsubstituted C1-C4 alkoxy, unsubstituted C 1-C4 haloalkyl, unsubstituted C1-C4 hydroxyalkyl, or unsubstituted C1-C4 haloalkoxy or hydroxy. Particular exemplaiy 'substituted 19 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 alkoxy' groups are-OCF3, -OCH2CF3, -OCH2Ph, -OCH2-cyclopropyl, -OCH2CH2OH, and OCH2CH2NMe2. [51) "Amino" refers to the radical -NH2. (52) "Substituted amino" refers to an amino group of the formula -N(R 38 )2 wherein R 38 is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstitued alkenyl, substituted or unsubstitued alkynyl, substituted or unsubstitued carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstitued heteroaryl, or an amino protecting group, wherein at least one of R38 is not a hydrogen. In certain embodiments, each R38 is independently selected from hydrogen, C1-C8 alkyl, C3-C8 alkenyl, C3-C8 alkynyl, C6-C10 aryl, 5-10 membered heteroaryl, 4-10 membered heterocyclyl, or C3-C10 cycloalkyl; or C1-Cs alkyl, substituted with halo or hydroxy; C3-Cs alkenyl, substituted with halo or hydroxy; C3-Cs alkynyl, substituted with halo or hydroxy, or-(CH2)r(CG-C10 aryl), -(CH2)t(5-10 membered heteroaryl), -(CH2)1(C3-C10 cycloalkyl), or-(CH2)t(4-10 membered heterocyclyl), wherein tis an integer between 0 and 8, each of which is substituted by unsubstituted C1-C4 alkyl, halo, unsubstituted C1-C4 alkoxy, unsubstituted C1-C4 haloalkyl, unsubstituted C1-C4 hydroxyalkyl, or unsubstituted C 1-C4 haloalkoxy or hydroxy; or both R 38 groups are joined to form an alkylene group. [53) Exemplary "substituted amino" groups include, but are not limited to, -NR39-C1-C8 alkyl, -NR39-{CH 2)i(C6-C10 aryl), -NR39-{CH2)t(5-10 membered heteroaryl), -NR39 - (CH2)t(CrC10 cycloalkyl), and-NRw-(CH2)1(4-10 membered heterocyclyl), wherein tis an integer from Oto 4, for instance 1 or 2, each R39 independently represents hydrogen or C1-Cg alkyl; and any alkyl groups present, may themselves be substituted by halo, substituted or unsubstituted amino, or hydroxy; and any aryl, heteroaryl, cycloalkyl, or heterocyclyl groups present, may themselves be substituted by unsubstituted C1-C4 alkyl, halo, unsubstituted C1-C4 alkoxy, unsubstituted C1-C4 haloalkyl, unsubstituted C1-C4 hydroxyalkyl, or unsubstituted C1- C4 haloalkoxy or hydroxy. For the avoidance of doubt the term 'substituted amino' includes the groups alkylamino, substituted alkylamino, alkylarylamino, substituted alkylarylamino, arylamino, substituted arylamino, dialkylamino, and substituted dialkylamino as defined below. Substituted amino encompasses both monosubstituted amino and disubstituted amino groups. [54) "Azido" refers to the radical -N3. 20 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[55] "Carbamoyl" or "amido" refers to the radical -C(O)NH2.

[56] "Substituted carbamoyl" or "substituted amido" refers to the radical -C(O)N(R62)2 wherein each R62 is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstitued alkenyl, substituted or unsubstitued alkynyl, substituted or unsubstitued carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or 1msubstituted aryl, substituted or unsubstitued heteroaryl, or an amino protecting group, wherein at least one ofR62 is not a hydrogen. In certain embodiments, R62 is selected from H, C1-C8 alkyl, C3-C10 cycloalkyl, 4-10 membered heterocyclyl, C6-C10 aryl, and 5-10 membered heteroaryl; or C1-Cg alkyl substituted with halo or hydroxy; or C3-C10 cycloalkyl, 4-10 membered heterocyclyl, C6- C10 aryl, or 5-10 membered heteroaryl, each of which is substituted by unsubstituted C 1-C4 alkyl, halo, m1substituted C1-C4 alkoxy, unsubstituted C1-C4 haloalkyl, unsubstituted C1-C4 hydroxyalkyl, or unsubstituted C 1-C4 haloalkoxy or hydroxy; provided that at least one R62 is other than H.

[57] "Carboxy" refers to the radical -C(O)OH.

[58] "Cyano" refers to the radical -CN.

[59] "Hydroxy" refers to the radical -OH.

[60] "Nitro" refers to the radical-N02.

[61] "Ethenyl" refers to substituted or unsubstituted-(C=C)-. "Ethylene" refers to substituted or unsubstituted-(C-C)-. "Ethynyl" refers to ---{C=C)-.

[62] "Nitrogen-containing heterocyclyl" group means a 4- to 7- membered non-aromatic cyclic group containing at least one nitrogen atom, for example, but without limitation, morpholine, piperidine ( e.g. 2-piperidinyl, 3-piperidinyl and 4-piperidinyl), pyrrolidine (e.g. 2- pyrrolidinyl and 3-pyrrolidinyl), azetidine, pyrrolidone, imidazoline, imidazolidinone, 2- pyrazoline, pyrazolidine, piperazine, and N-alkyl piperazines such as N-methyl piperazine. Particular examples include azetidine, piperidone and piperazone.

[63] Alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl groups, as defined herein, are optionally substituted (e.g., "substituted" or "unsubstituted" alkyl, "substituted" or "unsubstituted" alkenyl, "substituted" or "unsubstituted" alkynyl, "substituted" or "unsubstituted" carbocyclyl, "substituted" or "unsubstituted" heterocyclyl, "substituted" or "unsubstituted" aryl or "substituted" or "unsubstituted" heteroaryl group). In general, the term 21 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 "substituted'', whether preceded by the term "optionally" or not, means that at least one hydrogen present on a group ( e.g., a carbon or nitrogen atom) is replaced with a pennissible substituent, e.g., a substituent which upon substitution results in a stable compound, e.g., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, or other reaction. Unless otherwise indicated, a "substituted" group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituent is either the same or different at each position. The term "substituted" is contemplated to include substitution with all permissible substituents of organic compounds, any of the substituents described herein that results in the formation of a stable compound. The present invention contemplates any and all such combinations in order to arrive at a stable compound. For purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and / or any suitable substituent as described herein which satisfy the valencies of the heteroatoms and results in the formation of a stable moiety.

[64] Exemplary carbon atom substituents include, but are not limited to, halogen, -CN, - NO2, -N3, -SO2H, -SO3H, -OH, -ORa\-ON(Rbb)z, -N(Rbb)z,-N(Rbb) / X-, -N(ORcc)Rbb' - SH, -SRa\-SSRcC,-C(=O)Ra\-CO2H, -CHO, -C(ORcc)z, -CO2R33,-OC(=O)Ra3,-OCO2Ra\ -C(=O)N(Rbb)2, -OC(=O)N(Rbb)2, -NRbbC(=O)Raa, -NRbbC02Raa, -NRbbC(=O)N(Rbb)2, C(=NRbb)Raa, -C(=NRbb)ORaa, -OC(=NRbb)Raa, -OC(=NRbb)ORaa, -C(=NRbb)N(Rbb)2, - OC(=NRbb)N(Rbb)2, -NRbbC(=NRbb)N(Rbb)z, -C(=O)NRt,hS02Ra\-NRbbS02Raa, -S02N(Rbb)2, -SO2R33 , -SO2ORaa, -OSO2Raa, -S(=O)Ra\-OS(=O)Raa, -Si(R33)3, -OSi(R33)3-C(=S)N(Rbbh, -C(=O)SRaa, -C(=S)SRaa, -SC(=S)SR8 \ -SC(=O)SRaa, -OC(=O)SR3\-SC(=O)ORa\ - SC(=O)Raa, -P(=O)zRa\-OP(=O)2Raa, -P(=O)(Raa)2, -OP(=O)(Raa)2, -OP(=O)(ORcc)2, P(=0)2N(Rbb)2, -OP(=0)2N(Rbb)2, -P(=O)(NRbb)2, -OP(=O)(NRbb)2, -NRbbp(=O)(ORCC)2, - NRbbp(=O)(NRbb)2, -P(Rcc)2, -P(Rcc)3, -OP(Rcc)2, -OP(Rcc)3, -B(Raa)2, -B(ORcc)2, - BRaa(ORcc), C1-10 alkyl, C1-10 perhaloalkyl, C2-10 alkenyl, C2-10 alkynyl, C3-10 carbocyclyl, 3- 14 membered heterocyclyl, CG-14 aryl, and 5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups; 22 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 each instance ofRaa is, independently, selected from C1_1o alkyl, C1-10 perhaloalkyl, C2-io alkenyl, C210 alkynyl, C110 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Raa groups are joined to form a 3-14 membered heterocyclyl or 5- 14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups; each instance ofRbb is, independently, selected from hydrogen, -OH, -OR a\ -N(Rcc)2, - CN, -C(=O)Raa, -C(=O)N(Rcc)2, -CO2Raa, -SO2Raa, -C(=NRcc)OR8 \ -C(=NRcc)N(Rcc)2, SO2N(Rcc)i, -SO2RcC, -SO2ORc\ -SORaa, -C(=S)N(Rcc)2, -C(=O)SRcC, -C(=S)SRcC, - P(=O)2Raa, -P(=O)(R3a)2, -P(=O)2N(Rcc)2, -P(=O)(NRcc)i, C1-10 alkyl, C1-10 perhaloalkyl, C2-10 alkenyl, C2-10 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two R bb groups are joined to form a 3-14 membered heterocyclyl or 5- 14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups; each instance of Rec is, independently, selected from hydrogen, C1_10 alkyl, C1_10 perhaloalkyl, C2_10 alkenyl, C2_10 alkynyl, C3_10 carbocyclyl, 3-14 membered heterocyclyl, Cc,-14 aryl, and 5-14 membered heteroaryl, or two R cc groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups; each instance ofRdd is, independently, selected from halogen, -CN, -NO2, -N3, -SO2H, - SO 3H , -OH, -ORee , -ON(Rff),.,,.; ,,, -N(Rff) 2, -N(Rff) 3+ x-, -N(ORee)Rff, -SH, -SRee , -SSRee , C(=O)Ree, -CO2H, -CO2Ree, -OC(=O)Ree, -OCO2Ree, -C(=O)N(R11)2, -OC(=O)N(Rff)z, - NRffC(=O)Ree, -NRffC02Re\ -NRffC(=O)N(Rff)2, -C(=NRn)ORee, -OC(=NRff)Ree, - OC(=NRff)ORee, -C(=NRff)N(Rff)i, -OC(=NRff)N(Rff)2, -NRffC(=NRff)N(Rffh,~NRffS02Ree, - S02N(Rrr)i, -SO2Re\ -SO2ORee, -OSO2Re\-S(=O)Ree, -Si(Ree)3, -OSi(Ree)3, -C(=S)N(Rf1)2, - C(=O)SRee, -C(=S)SRee, -SC(=S)SRee, -P(=O)2Ree, -P(=O)(Ree)i, -OP(=O)(Ree)2, - OP(=O)(ORee)2, C1-6 alkyl, C1-6 perhaloalkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 carbocyclyl, 3-10 membered heterocyclyl, C6_10 aryl, 5-10 membered heteroaryl, wherein each alkyl, alkenyl, 23 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups; each instance of Ree is, independently, selected from C1--6 alkyl, C1-6 perhaloalkyl, C2--6 alkenyl, C2_6 alkynyl, C3-1o carbocyclyl, C6_10 aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups; each instance ofRff is, independently, selected from hydrogen, C1-6 alkyl, C1-6 perhaloalkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 carbocyclyl, 3-10 membered heterocyclyl, C6-IO aryl and 5-10 membered heteroaryl, or two R ff groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R gg groups; and each instance ofRgg is, independently, halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OC1-6 alkyl, -ON(C1-6 alkyl)2, -N(C1-6 alkyl)2, -N(C1-6 alkyl) / X-, -NH(C1-6 alkyl) / X-, - NH2(C1-6 alkyl) +x-, -NH / X-, -N(OC1-6 alkyl)(C1-6 alkyl), -N(OH)(C1-6 alkyl), -NH(OH), - SH, -SCl-6 alkyl, -SS(C1_6 alkyl), -C(=O)(C1-6 alkyl), -CO2H, -CO2(C1_6 alkyl), -OC(=O)(C1-6 alkyl), -OCO2(C1--0 alkyl), -C(=O)NH2, -C(=O)N(C1--0 alkylh, -OC(=O)NH(C1--0 alkyl), - NHC(=O)( C1-6 alkyl), -N(C1-6 alkyl)C(=O)( Cl--6 alkyl), -NHCO2(C1-6 alkyl), -NHC(=O)N(C1- 6 alkyl)2, -NHC(=O)NH(C1-6 alkyl), -NHC(=O)NH2, -C(=NH)O(C1-6 alkyl),-OC(=NH)(C1-o alkyl), -OC(=NH)OC1-6 alkyl, -C(=NH)N(C1_6 alkyl)2, -C(=NH)NH(C1-6 alkyl), -C(=NH)NH2, -OC(=NH)N(C1-6 alkyl)2, -OC(NH)NH(Ct-6 alkyl), -OC(NH)NH2, -NHC(NH)N(Ct-6 alkyl)2, NHC(=NH)NH2, -NHSO2(C1-6 alkyl), -SO2N(C1-6 alkyl)2, -SO2NH(Ci-6 alkyl), -SO2NH2,SO2C1- 6 alkyl, -SO2OC1-6 alkyl, -OSO2C1-6 alkyl, -SOC1-6 alkyl, -Si(C1-6 alkyl)3, -OSi(C1-o alkyl)3 -C(=S)N(C1-6 alkyl)2, C(=S)NH(C1-6 alkyl), C(=S)NH2, -C(=O)S(C1-6 alkyl), - C(=S)SC1-6 alkyl, -SC(=S)SC1-6 alkyl, -P(=O)2(C1-6 alkyl), -P(=O)(C1-6 alkyl)2, -OP(=O)(C1-6 alkyl)z, -OP(=O)(OC1-6 alkylh, C1-6 alkyl, C1-6 perhaloalkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 carbocyclyl, C6_10 aryl, 3-10 membered heterocyclyl, 5-10 membered heteroaryl; wherein x- is a counterion. 24 Date Rec;ue / Date Received 2024-04-12 84019316

[65] A "counterion" or "anionic counterion" is a negatively charged group associated with a cationic quaternary amino group in order to maintain electronic neutrality. Exemplary counterions include halide ions (e.g., F-, er, Bi-, r), NO3 , ClO4 , Off, H2PO4 , HSO4 , sulfonate ions (e.g., methansulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10-camphor sulfonate, naphthalene---2-sulfonate, naphthalene---1-sulfonic acid-5-sulfonate, ethan-1-sulfonic acid-2-sulfonate, and the like), and carboxylate ions (e.g., acetate, ethanoate, propanoate, benzoate, glycerate, lactate, tartrate, glycolate, and the like). [ 66) Nitrogen atoms can be substituted or unsubstituted as valency pem1its, and include primary, secondary, tertiary, and quartemary nitrogen atoms. Exemplary nitrogen atom substitutents include, but are not limited to, hydrogen, -OH, -OR a\ -N(Rcc)2, -CN, -C(=O)Raa, -C(=O)N(Rcc)2, -CO2Raa, -SO2Ra\ -C(=NRbb)Ra\ -C(=NRcc)ORaa, -C(=NRcc)N(Rcc)2, - SO2N(Rcc)2, -SO2RcC, -SO2ORcC, -SORaa, -C(=S)N(Rcc)2, -C(=O)SRcC, -C(=S)SRcc, - P(=O)zRaa, -P(=O)(Raa)z, -P(=O)2N(Rcc)2, -P(=O)(NRcch, C1-10 alkyl, C1-10 perhaloalkyl, C2-10 alkenyl, C2-10 alkynyl, C3_10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14- membered heteroaryl, or two R cc groups attached to a nitrogen atom are joined to fom1 a 3-14- membered heterocyclyl or 5-14-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Reid groups, and wherein Ra\ Rb\ Rec and Rdd are as defined above. [ 67) In certain embodiments, the substituent present on a nitrogen atom is an amino protecting group (also referred to herein as a nitrogen protecting group). Amino protecting groups include, but are not limited to, -OH, -OR88 , -N(Rcc)2, -C(=O)Raa, -C(=O)OR08 , - C(=O)N(Rcc)2, -S(=O)2R88 , -C(=NRcc)Raa, -C(=NRcc)ORa\ -C(=NRcc)N(Rcc)z, -SO2N(Rcc)z, - SO2Rcc, -SO2ORc\-SOR08,-C(=S)N(Rcc)2, -C(=O)SRc\-C(=S)SRcc, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-10 carbocyclyl, 3-14-membered heterocyclyl, C6-14 aryl, and 5-14- membered heteroaryl groups, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rrld groups, and wherein R0 \ Rb\ Rec and Reid are as defined herein. Amino protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic .S:Vnthesis, T. W. Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999. 25 Date Rec;ue / Date Received 2024-04-12 84019316

[68] Exemplary amino protecting groups include, but are not limited to amide groups (e.g., - C(=O)Raa), which include, but are not limited to, formamide and acetamide; carbamate groups (e.g .. -C(=O)ORaa), which include, but are not limited to, 9-fluorenylmethyl carbamate (Fmoc), t-butyl carbamate (BOC), and benzyl carbamate (Cbz); sulfonamide groups (e.g., -S(=O)2Raa), which include, but are not limited to,p-toluenesulfonamide (Ts), methanesulfonamide (Ms), and N-[2-(trimethylsilyl)ethoxy]methylan1ine (SEM).

[69] In certain embodiments, the substituent present on an oxygen atom is an oxygen protecting group (also referred to as a hydroxyl protecting group). Oxygen protecting groups include, but are not limited to, -Raa, -N(Rbb)2, -C(=O)SRa\ -C(=O)Ra\-CO2Ra\ _ C(=O)N(Rbb)2, -C(=NRbb)Ra\-C(=NRbb)ORa\-C(=NRbb)N(Rbb)2, -S(=O)Raa, -SO2Ra\Si( Rm\)3_-P(Rcc)2, -P(Rcc)3, -P(=O)zRl1ll, -P(=O)(Raa)i, -P(=O)(ORcc)i, -P(=O)2N(Rbb)2, andP(= O)(NRbb)2, wherein Raa, Rbb' and Rec are as defined herein. Oxygen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999.

[70] Exemplary oxygen protecting groups include, but are not limited to, methyl, methoxylmethyl (MOM), 2-methoxyethoxymethyl (MEM), benzyl (Bn), triisopropylsilyl (TIPS), t-butyldimethylsilyl (TBDMS), t-butylmethoxyphenylsilyl (TBMPS), methanesulfonate (mesylate), and tosylate (Ts).

[71] In certain embodiments, the substituent present on an sulfur atom is an sulfur protecting group (also referred to as a thiol protecting group). Sulfur protecting groups include, but are not limited to, -Raa, -N(Rbb)2, -C(=O)SRa\ -C(=O)Ra\ -CO2Ra\ -C(=O)N(Rbb)z, -C(=NRbb)Raa, - C(=NRt,h)ORaa, -C(=NRt,h)N(Rt,h)2, -S(=O)Raa, -SO2Raa, -Si(Raah-P(Rcc)2, -P(Rcc)3, - P(=O)2Raa, -P(=O)(Raa)2, -P(=O)(ORcc)2, -P(=O)zN(Rbb)z, and-P(=O)(NRt,h )2, wherein Raa, Rbb' and Rec are as defined herein. Sulfur protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999. [72) These and other exemplary substituents are described in more detail in the Detailed Description, and Examples. The invention is not intended to be limited in any manner by the above exemplary listing of substituents. 26 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Other definitions

[73] As used herein, the term "modulation" refers to the inhibition or potentiation of GABA receptor function. A "modulator" (e.g., a modulator compound) may be, for example, an agonist, partial agonist, antagonist, or partial antagonist of the GABA receptor. (74] "Pham1aceutically acceptable" means approved or approvable by a regulatory agency of the Federal or a state government or the corresponding agency in countries other than the United States, or that is listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals, and more particularly, in humans. (75] "Pharmaceutically acceptable salt" refers to a salt of a compound of the invention that is pharmaceutically acceptable and that possesses the desired phannacological activity of the parent compound. In particular, such salts are non-toxic may be inorganic or organic acid addition salts and base addition salts. Specifically, such salts include: (1) acid addition salts, formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or formed with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyrnvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane---disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-l-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like; or (2) salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base such as ethanolamine, diethanolamine, triethanolamine, Nmethylglucamine and the like. Salts further include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and when the compound contains a basic functionality, salts of non-toxic organic or inorganic acids, such as 27 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate and the like. The term "pharmaceutically acceptable cation" refers to an acceptable cationic counter-ion of an acidic fonctional group. Such cations are exemplified by sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium cations, and the like. See, e.g., Berge, et al., J. Phann. Sci. (1977) 66(1): 1-79.

[76] "Solvate" refers to forms of the compound that are associated with a solvent or water (also referred to as "hydrate"), usually by a solvolysis reaction. This physical association includes hydrogen bonding. Conventional solvents include water, ethanol, acetic acid, and the like. The compounds of the invention may be prepared e.g. in crystalline form and may be solvated or hydrated. Suitable solvates include pharmaceutically acceptable solvates, such as hydrates, and further include both stoichiometric solvates and non-stoichiometric solvates. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. "Solvate" encompasses both solution-phase and isolable solvates. Representative solvates include hydrates, ethanolates and methanolates.

[77] As used herein, the tem1 "isotopic variant" refers to a compound that contains unnatural proportions of isotopes at one or more of the atoms that constitute such compound. For example, an "isotopic variant" of a compound can contain one or more non-radioactive isotopes, such as for example, deuterium (2H or D), carbon-13 ( 13C), nitrogen-15 ( 15N), or the like. It will be understood that, in a compound where such isotopic substitution is made, the following atoms, where present, may vary, so that for example, any hydrogen may be 2H / D, any carbon may be 13C, or any nitrogen may be 15N, and that the presence and placement of such atoms may be detennined within the skill of the art. Likewise, the invention may include the preparation of isotopic variants with radioisotopes, in the instance for example, where the resulting compounds may be used for drng and / or substrate tissue distribution studies. The radioactive isotopes tritium, i.e., 3H, and carbon-14, i.e., 14C, are particularly usefol for this purpose in view of their ease of incorporation and ready means of detection. Further, compounds may be prepared that are substituted with positron emitting isotopes, such as 11C, 18F, 150, and 13N, and would be useful in Positron Emission Topography (PET) studies for 28 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 examining substrate receptor occupancy. All isotopic variants of the compounds provided herein, radioactive or not, are intended to be encompassed within the scope of the invention. [78) "Stereoisomers": It is also to be understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed "isomers." Isomers that differ in the arrangement of their atoms in space are termed "stereoisomers." Stereoisomers that are not mirror images of one another are termed "diastereomers" and those that are nonsuperimposable mirror images of each other are tem1ed "enantiomers." When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute confif,>uration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e., as(+) or (-)-isomers respectively). A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a "racemic mixture". [79) "Tautomers" refer to compounds that are interchangeable forms of a particular compound structure, and that vary in the displacement of hydrogen atoms and electrons. Thus, two structures may be in equilibrium through the movement of n electrons and an atom (usually H). For example, enols and ketones are tautomers because they are rapidly interconverted by treatment with either acid or base. Another example of tautomerism is the aci- and nitro- forms of phenylnitromethane, that are likewise formed by treatment with acid or base. Tautomeric forms may be relevant to the attainment of the optimal chemical reactivity and biological activity of a compound of interest. [80) A "subject" to which administration is contemplated includes, but is not limited to, humans (i.e., a male or female of any age group, e.g., a pediatric subject (e.g, infant, child, adolescent) or adult subject (e.g .. young adult, middle-aged adult or senior adult)) and / or a nonhuman animal, e.g., a mammal such as primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In certain embodiments, the subject is a human. In certain embodiments, the subject is a non-human animal. The terms "human," "patient," and "subject" are used interchangeably herein. 29 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[81] Disease, disorder, and condition are used interchangeably herein.

[82] As used herein, and unless otherwise specified, the terms "treat," "treating" and "treatment" contemplate an action that occurs while a subject is suffering from the specified disease, disorder or condition, which reduces the severity of the disease, disorder or condition, or retards or slows the progression of the disease, disorder or condition ("therapeutic treatment"), and also contemplates an action that occurs before a subject begins to suffer from the specified disease, disorder or condition ("prophylactic treatment").

[83] In general, the "effective amount" of a compound refers to an amount sufficient to elicit the desired biological response, e.g., to treat a CNS-related disorder, is sufficient to induce anesthesia or sedation. As will be appreciated by those of ordinary skill in this art, the effective amount of a compound of the invention may vary depending on such factors as the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, weight, health, and condition of the subject. An effective amount encompasses therapeutic and prophylactic treatment.

[84] As used herein, and unless otherwise specified, a "therapeutically effective amount" of a compound is an amount sufficient to provide a therapeutic benefit in the treatment of a disease, disorder or condition, or to delay or minimize one or more symptoms associated with the disease, disorder or condition. A therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapies, which provides a therapeutic benefit in the treatment of the disease, disorder or condition. The term "therapeutically effective amount" can encompass an amount that improves overall therapy, reduces or avoids symptoms or causes of disease or condition, or enhances the therapeutic efficacy of another therapeutic agent.

[85] As used herein, and unless otherwise specified, a "prophylactically effective amount" of a compound is an amount sufficient to prevent a disease, disorder or condition, or one or more symptoms associated with the disease, disorder or condition, or prevent its recurrence. A prophylactically effective amount of a compound means an amount of a therapeutic agent, alone or in combination with other agents, which provides a prophylactic benefit in the prevention of the disease, disorder or condition. The term "prophylactically effective amount" can encompass 30 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 an amount that improves overall prophylaxis or enhances the prophylactic efficacy of another prophylactic agent. Detailed Description of Certain Embodiments of the Invention (86] As generally described herein, the present invention provides C21-substituted neuroactive steroids designed, for example, to act as GABA modulators. In certain embodiments, such compounds are envisioned to be useful as therapeutic agents for the inducement of anesthesia and / or sedation in a subject. In certain embodiments, such compounds are envisioned to be useful as therapeutic agents for treating a CNS-related disorder. Compound~

[87] In one aspect, provided is a compound of Formula (I): HQ\\' R2 (I), a phannaceutically acceptable salt thereof, wherein: A is an optionally substituted nitrogencontaining heteroaryl or heterocyclyl; Lis -C(R3)(R3 )-, -0-, -S-, or -NR3 -; R1 is hydrogen or C 1-C6 alkyl, C 1-C6 alkenyl, C 1-C6 alkynyl, carbocyclyl, or heterocyclyl; R 2 is hydrogen, C 1-C6 alkyl (e.g., C1-C6 haloalkyl), or C1-C6 alkoxy; each R3 is independently hydrogen or C1-C6 alkyl; R5 is absent or hydrogen; and ::..:..:.:.:..: represents a single or double bond, wherein when one of ::..:..:.:.:..: is a double bond, the other ::.::..::..:..: is a single bond; and when one of the ::.::..::..:..: is a double bond, R5 is absent.

[88] In one aspect, provided is a compound of Fommla (Ia): 31 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 (Ia), a pham1aceutically acceptable salt thereof, wherein: A is an optionally substituted nitrogencontaining heteroaryl or heterocyclyl; Lis -C(R3)(R3 )-, -0-, -S-, or -NR3 -; R1 is hydrogen or C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, carbocyclyl, or heterocyclyl; each R3 is independently hydrogen or C1-C6 alkyl; R5 is absent or hydrogen; and :.::.::.::.:.: represents a single or double bond, wherein when one of :.::.::.::.:.: is a double bond, the other :.::.::.::.:.: is a single bond; and when one of the :.::.::.::.:.: is a double bond, R5 is absent.

[89] In some embodiments, the compotmd is of the Formula (Ia-1): (Ia-1). (90] In some embodiments, the compound is of the Formula (Ia-2): (Ia-2).

[91] In some embodiments, A is monocyclic or bicyclic. 32 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[92] In some embodiments, A is monocyclic. In some aspects of these embodiments, A is attached through a nitrogen. In some embodiments, A is a heteroaryl. In some aspects of these embodiments, the heteroaryl comprising up to five nitrogen atoms. In some embodiments, A is a 5-membered heteroaryl or heterocyclyl. In some embodiments, A is a 5-membered heteroaryl or heterocyclyl comprising up to four nitrogen atoms. In some embodiments, A is a 5- membered heteroaryl or heterocyclyl comprising 2, 3, or 4 nitrogen atoms. In some embodiments, A is pyrazole, triazole, or tetrazole. In some embodiments, A is unsubstituted pyrazole, triazole, or tetrazole. N~ 'N -NI / In some embodiments, A is 1< , n N-N or 1< ("' N~N, some embodiments, A is unsubstituted triazole. In some embodiments, A is llJ,N In [93) In some embodiments, A is bicyclic. In some aspects of these embodiments, A is attached through a nitrogen. In some embodiments, A is a heteroaryl. In some aspects of these embodiments, the heteroaryl comprises up to five nitrogen atoms. In some embodiments, the heteroaryl comprises at least two nitrogen atoms. In some embodiments, A is a heteroaryl comprising up to four nitrogen atoms. In some embodiments, A is a heteroaryl comprising up to three nitrogen atoms. In some embodiments, A is a heteroaryl comprising 2, 3, or 4 nitrogen atoms. In some embodiments, the heteroaryl is benzotriazole, azabenzotriazole, diazabenzotriazole, benzopyrazole, azabenzopyrazole, or diazabenzopyrazole. [94) In some embodiments, the compound is of the Formula (Ia-3): 0(R'), N (Ia-3) 33 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 wherein R4 is cyano, nitro, hydroxy, halo, C1-C6 alkyl, C1-C6 alkoxy, -C(O)R\-C(O)N(Rb)(Rc), ---C(O)ORa, -N(Rb)(Rc), -OC(O)N(R1)(Rc), -OC(O)ORa, -OC(O)R\ -S(O)o-2R\-S(O)o-2ORa, or -S(O)0_2N(Rb)(Rc); each Ra is hydrogen or C1-C6 alkyl; each Rb and Re is independently hydrogen, C 1_C6 alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, or Rh and RC, together with the nitrogen atom to which they are bound to form a ring (e.g., a 3-7-membered ring, e.g., a 5-7- membered ring; a ring containing at least one heteroatom, e.g., a nitrogen, oxygen, or sulfur atom); and n is 0, 1, 2, or 3. [95) In some embodiments, the compound is of the Formula (Ia-4): 0(R'la H (Ia-4) wherein R4 is cyano, nitro, hydroxy, halo, C1-C6 alkyl, C1-C6 a1koxy, -C(O)R'\-C(O)N(Rb)(Rc), ---C(O)OR\-N(Rb)(Rc), -OC(O)N(Rb)(Rc), -OC(O)ORa, -OC(O)Ra, -S(O)o-2Ra, -S(O)o-2OR\ or -S(O)o-2N(Rb)(Rc); each Ra is hydrogen or C1-C6 alkyl; each Rb and Re is independently hydrogen, C1-C6 alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, or Rb and R°, together with the nitrogen atom to which they are bound to form a ring ( e.g., a 3-7-membered ring, e.g., a 5-7- membered ring; a ring containing at least one heteroatom, e.g., a nitrogen, oxygen, or sulfur atom); and n is 0, 1, 2, or 3. (96] In some embodiments, n is 0.

[97] In some embodiments, 11 is 1. In some aspects of these embodiments, R4 is cyano, nitro, hydroxy, halo, C1-C6 alkyl, C1-C6 alkoxy, -C(O)Ra, ---C(O)OR\ or-S(O)o-2Ra. In some embodiments, n is l and R4 is cyano, nitro, hydroxy, halo, C1-C6 alkyl, C1-C6 alkoxy, -C(O)R\ -C(O)ORa, or-S(O)o_2Ra_ In some aspects of these embodiments, R4 is halo (e.g., F, Cl, Br). In some embodiments, R4 is F. In some embodiments, R4 is CL In some embodiments, R4 is Br. In some embodiments, R4 is C1-C6 alkoxy (e.g., -OCH3, -OCH2CH3). In some embodiments, R4 is cyano. In some embodiments, R4 is -C(O)Ra or-C(O)ORa_ In some aspects of these 34 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 embodiments, Ra is C1-C6 alkyl (e.g., -CH3, -CH2CH3). In some embodiments, R4 is -S(0)o_2Ra_ In some embodiments, R4 is -S(0)2Ra, and Ra is C1-C6 alkyl. In some aspects of these embodiments, Ra is -CH3. In some embodiments, R4 is -S(OhCH3. In some embodiments, R4 is C1-C6 alkyl (e.g., -CH3, -CH2CH3).

[98] In some embodiments, n is 2. In some aspects of these embodiments, R4 is cyano, nitro, hydroxy, halo, C1-C6 alkyl, C1-C6 alkoxy, -C(0)Ra,-C(0)0R\ or-S(0)o-2Ra. In some embodiments, n is 2 and R4 is cyano, nitro, hydroxy, halo, C1-C6 alkyl, C1-C6 alkoxy, -C(0)R\ -C(0)0Ra, or-S(0)0_2R3 . In some aspects of these embodiments, R4 is halo (e.g., F, Cl, Br). In some embodiments, R4 is F. In some embodiments, R4 is CL In some embodiments, R4 is Br. In some embodiments, R4 is C1-C6 alkoxy (e.g., -0CH3, -0CH2CH3). In some embodiments, R4 is cyano. In some embodiments, R4 is -C(0)Ra or-C(0)0Ra. In some aspects of these embodiments, Ra is C1-C6 alkyl (e.g., -CH3, -CH2CH3). In some embodiments, R4 is -S(0)0_2Ra. In some embodiments, R4 is -S(0)2R\ and Ra is C1-C6 alkyl. In some aspects of these embodiments, Ra is -CH3. In some embodiments, R4 is C1-C6 alkyl (e.g., -CH3, -CH2CH3).

[99] In some embodiments, n is 1 or 2, and R4 is C 1-C6 alkyl or C(0)0Ra. In some embodiments, the compound R4 is methyl. In some embodiments, the compound Ra is C1-C6 alkyl. In some embodiments, Ra is ethyl.

[100] In some embodiments, R1 is hydrogen or C1-C6 alkyl, and R4 is-C(0)0Ra.

[101] n some embodiments, R1 is C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, carbocyclyl, or heterocyclyl. In some embodiments, the compound R 1 is C 1-C6 alkyl. In some embodiments, the compound R 1 is methyl, ethyl, or isopropyl.

[102] In some embodiments, R1 is methyl and R4 is-C(0)0Et. In some embodiments, the compound is selected from: 35 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227

[103] In one aspect, provided is a compound of Formula (lb): Date Rec;ue / Date Received 2024-04-12 H0\1' R2 36 00 L: (lb), PCT / US2014 / 052417 WO 2015 / 027227 PCT / US2014 / 052417 a pham1aceutically acceptable salt thereof, wherein: A is an optionally substituted nitrogencontaining heteroaryl or heterocyclyl; Lis -C(R3)(R3 )-, -0-, -S-, or-NR3 -; R1 is hydrogen or C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, carbocyclyl, or heterocyclyl; R2 is C1-C6 alkyl (e.g., C1-C6 haloalkyl) or C1-C6 alkoxy; each R3 is independently hydrogen or C1-C6 alkyl; R5 is absent or hydrogen; and ::.:..:..:..:.. represents a single or double bond, wherein when one of ::.:..:..:..:.. is a double bond, the other ::.:..:..:..:.. is a single bond; and when one of the ::.:..:..:..:.. is a double bond, R5 is absent.

[104] In some embodiments, the compound is of the Formula (Ib-1): HQ\\' R2 (Ib-1).

[105] In some embodiments, the compound is of the Formula (Ib-2): HQ\1' R2 H (Ib-2).

[106] In some embodiments, the compound is of the Formula (lb-3) 37 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 HO\'' R2 PCT / US2014 / 052417 (Ib-3), wherein R4 is cyano, nitro, hydroxy, halo, C1-C6 alkyl, C1-C6 alkoxy, -C(O)Ra, -C(O)N(Rb)(Rc), -C(O)OR\ -N(Rb)(Rc), -OC(O)N(Rb)(Rc), -OC(O)ORa, -OC(O)R3, -S(O)o_2R\-S(O)o-2OR\ or -S(O)0_2N(Rh)(Rc); each Ra is hydrogen or C1-C6 alkyl; each Rh and Re is independently hydrogen, C1-C6 alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, or Rb and RC, together with the nitrogen atom to which they are bound to form a ring (e.g., a 3-7-membered ring, e.g., a 5-7- membered ring; a ring containing at least one heteroatom, e.g., a nitrogen, oxygen, or sulfur atom); and n is 0, 1, 2, or 3.

[107] In some embodiments, the compound is of the Formula (Ib-4) HQ\\I R2 H 0("')" N (lb-4), wherein R4 is cyano, nitro, hydroxy, halo, C1-C6 alkyl, C1-C6 alkoxy, -C(O)R3,-C(O)N(Rb)(Rc), -C(O)OR\ -N(Rb)(Rc), -OC(O)N(Rb)(Rc), -OC(O)ORa, -OC(O)Ra, -S(O)o-2Ra, -S(O)o-2OR3, or -S(O)0_2N(Rb)(Rc); each Ra is hydrogen or C1-C6 alkyl; each Rb and Re is independently hydrogen, C 1_C6 alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, or Rb and RC, together with the nitrogen atom to \vhich they are bound to fom1 a ring (e.g., a 3-?-membered ring, e.g., a 5-?membered ring; a ring containing at least one heteroatom, e.g., a nitrogen, oxygen, or sulfur atom); and n is 0, 1, 2, or 3.

[108] In some embodiments, A is monocyclic. 38 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[109] In some embodiments, A is bicyclic.

[110] In some embodiments, A is attached through a nitrogen.

[111] In some embodiments, A is a 5-membered or 6-membered heteroaryl or heterocyclyl. In some embodiments, A is a 5-membered or 6-membered heteroaryl or heterocyclyl comprising up to four nitrogen atoms. In some embodiments, A is a 5-membered or 6-membered heteroaryl orheterocyclyl comprising 1, 2, 3, or4 nitrogen atoms.

[112] In some embodiments, A is a heterocyclyl. In some embodiments, A is morpholine or H (o) (N) N N p1perazme. In some embodiments, A is .,L or ~ .

[113] In some embodiments, A is a heteroaryl. In some aspects of these embodiments, the heteroaryl comprises up to five nitrogen atoms. In some embodiments, the heteroaryl is benzotriazole, azabenzotriazole, diazabenzotriazole, benzopyrazole, azabenzopyrazole, or / QP LN N-N \_.J \_,; diazabenzopyrazole. In some embodiments, A is ,,. ,,. 7n ~ N-N ' , or p I \ N-N y

[114] In some embodiments, the heteroaryl is 5-membered. ;? N1 / n \~ N-N N-N X '< r1~ ~ N-N <

[115] In some embodiments, A comprises up to four nitrogen atoms. In some embodiments, A comprises 2, 3, or 4 nitrogen atoms. In some embodiments, A is pyrazole, triazole, or tetrazole. In some embodiments, A is unsubstituted pyrazole, triazole, or tetrazole. In some embodiments, 39 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 N°N \ ,, <N, ,,, N N-N N-N" Ais X X unsubstituted triazole. N0-_ ' I / N-N X n N-N ,or X r N-N, . . II ;,N In some embodunents, A 1s "--!I PCT / US2014 / 052417 Iu some embodiments, A is [116) In some embodiments, R1 is C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, carbocyclyl, or heterocyclyl. In some embodiments, R1 is C1-C6 alkyl. In some embodiments, R1 is methyl, ethyl, or isopropyl. In some embodiments, R 1 is methyl. In some embodiments, R 1 is ethyl.

[117] In some embodiments, R2 is C1_C6alkyl. In some embodiments, R2 is methyl. In some embodiments, R2 is C1.C6haloalkyl. In some embodiments, R2 is -CF3. [118) In some embodiments, n is 0.

[119] In some embodiments, n is I or 2, and R 4 is cyano, halo, C 1-C6 alkyl, C 1-C6 alkoxy, C(O)R\ or -S(O)o.2Ra.

[120] In some embodiments, R4 is Br, Cl, or F. In some embodiments, R4 is F. In some embodiments, R4 is Cl. In some embodiments, R4 is Br. In some embodiments, R4 is-OCH3. In some embodiments, R4 is -OCF3. In some embodiments, R4 is cyano. In some embodiments, R4 is C1-C6 alkyl. In some embodiments, R4 is methyl. In some embodiments, R4 is-C(O)Ra. In some embodiments, Ra is C1-C6 alkyl. In some embodiments, Ra is methyl. In some embodiments, R4 is -CF3. In some embodiments, R4 is-S(O)2Ra_ In some embodiments, Ra is methyl.

[121] In some embodiments, R1 is C1-C6 alkyl and R4 is -C(O)Ra_ In some embodiments, R1 is methyl and R4 is -C(O)Me. [122) In some embodiments, 11 is 2, and R4 is cyano, halo, C1-C6 alkyl, C1-C6 alkoxy, - C(O)Ra, or -S(O)0_2Ra. In some embodiments, n is 2, one R4 is F and one R4 is F. In some embodiments, n is 2, one R4 is F and one R4 is CL In some embodiments, n is 2, one R4 is - OCH3 and one R4 is -OCH3. [123) In some embodiments, n is O or 1; R1 is hydrogen or C1-C6 alkyl; and R2 is methyl. [124) In some embodiments, R1 is methyl, ethyl, or isopropyl. In some embodiments, R1 is methyl. 40 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[125] In some embodiments, R4 is C1-C6 alkyl, -C(O)Ra, or -S(O)o-2Ra_ In some embodiments, R4 is methyl. In some embodiments, R4 is -C(O)Me. In some embodiments, R4 is -S(O)2Me. In some embodiments, R4 is cyano. (126] In some embodiments, the compound is selected from: C) N CH3 0 I .::::s:o c) N Date Rec;ue / Date Received 2024-04-12 CH3 c) N 41 C) N WO 2015 / 027227 PCT / US2014 / 052417 H3C N-N N--N N--N y I \\ l ,, N~N :--.-.N / N CN H 0 N-{ N--N / N I ;>- N..::::-N / N..::::-N Hd . H 0 N:>--- N--N ~:--.-. Cl V-cN Hd H F ~:>- N~ N:--.-. ~N 42 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 N:>- N-N ~::,,.__ SO2Me , )>-_ N,N Hd - Hd- - A A 0 0 N-i EtO N,N,N ~N~ :>-CN Hd . A H 0 N:>- N--N Meo ~~ Me I )>- N.:::-N Hd H H N-{ N--N / N MeO / J N.:::-N / N~ H Hd H N--N N:>- ~ \ N Meo ~~ SO2Me H Hd H ~N~: >-Cl 43 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 0 0 ,N~-~ Nf_)CI N~ N \ / 7 'N ~ I Cl HCf A HO A 0 ~:0 / . NN,N \j )_ ,,1 / Cl N ,.__ N HO Ho' - A A 0 ~v ~~---~ N~N N ;:-._ ::--.. \ Ho' - A F ~:-0 N~N ~v· N~N N---N ~N 44 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 ~SJ ~:uJ N~N N~ N F HCf - A ~D--~ F ~0--~ N~ \ N~ \ ::--. ::--. OMe OMe F F ~:-0 ~{) N~N N~N Hcf - A 0 ~:-o-J F N~ N F N~~u ---\~ F ::--. F Hcf A F ~{J--~ F ~:OJ N~ \ N~ N F ::--. Hcf - F A N---N ~:Q-1 N;D~ O\M e N, 'N OMe ::--. Hcf - A 45 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Cl ~j:) N~N ~:-0 / ; N, 'N Cl HCf - H Meo 0Me ~~--~ Cl ~ N~ \ N~N ::--.. Hd- : H ~s~ / ; OMe N--N N, ;OOMe 'N 0Me N~ \ ::--.. 0Me Hd - H 0Me N----N ~:0 / ; ;~OMe Me0 N~ \ N, 'N 0Me ::--.. 0Me Hd- . A Br N~~: 0 / N, ~ N.--N \ Cl Hd H 46 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 0 ~vCI 0 EtO EtO ~0-~ N~N N :---.. \ ::--... Cl Ho' H Hd H 0 0 Meo N-N Meo NJ:JF N'" tl N' N ~ / ? ~' Hd F H HC{ H 0 MeO NV ~0-~ t-J_, \ / ? F N:---.. \ N ::--... Cl Ho' H H F. F MeO ~v Meo ~:0- / ; F N~N N~ N F Hd H Hd H ~0-~ F NV / ; N~ \ ~ N ::--... F H H 47 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Meo ~-0 N~N ~v / ; N~ N F H◊- H H 0 0 ~~---~ F N-N Meo N::--.. \ I~ N:--.. I ::---- ::.:::-._ F H◊- H◊- : H A F 0 F 0 I N~-o-: F N-N 0 I~ 'N N:--..:::,..__ ~ H◊- A H◊- H Cl 0 0 Cl j)- N ~-v~ N~ Cl I 0 N,, N N N H◊- : H H◊- H 0 0 :j) NN:-,,. ~N NJ H◊- H H◊- H 0 F 0 I 0--f-F N-N ~r,V F 0 N':--..t )l F :::,..__ \ o)<~ N H◊- H 48 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 0 0 ~-o--~ F F ,t.j)--c, OXF I 0 N =-- 'N Hd Hd A 0 0 N~~ u N:0-• ~ I Cl Hd A Hd H 0 0 ~u N-N I~ N:--. N:--. I \ ~ F ~ F Hd A Hd A Cl 0 F 0 N~~ :(;-: N~~ ~ I ::::::,... Hd A Hd H F 0 Cl 0 I ~:(;-: ~~ 0 N~ N~ \ ~ Hd A Hd A Cl F. F ~x,1 F ~v N" N.:-N N F 49 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 0 ~-{J_ / ~F N-N I \\ N" \ NIN :::c-- F Hd A 0 0 I N-( I ~y 0 ' N 0 N~ ,, N~ N 11 - N A 0 Cl 0 I ~-o-: I N-N 0 0 N~ N'~& N ~ \ Cl - - A A 0 0 F I ~tiJ--c1 I ~f:;-: 0 0 N~ N N F H 0 0 F. I N-N I N~i)--: 0 F I~ 0 N~ \ 'N ~ F - F A 50 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 0 0 I f-Jx I 0 0 f-J:(?~ N~ N,, N F F F 0-+F - F - F A A 0 0 0 N-N I Y' y I N:(? 0 N;-,_ N 0 CN NJ H H Cl y ~:j:5 / J N:--. CN H H 0 ~~~ N-N I 0 ~CN N" N F H H N-N F I N-N v-{-F I ,, 0 N.::::v N F H H ~xi / F ~tf--~ N::;- F N:--. ~ N F F ::--.. F F F H H 51 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 r~--; r=-0-J o\ N~ ::::---. ~ N.;:::- N F F F / 0 F H H 0- I 0 Nr.;:::-:¢§ r~--~ N N~ ::::---. ~ F F F F OH' / 0 H H I ~~---~ I ~{)_---~ 0 N::,-.. ~ 0 N::,-.. ~ F ::::---. Cl F ::::---. Cl F F Hcf H Hcf H Cl I ~:P I ~:v-CI 0 N~N 0 N~N F F F F Hcf H H 52 Date Rec;ue / Date Received 2024-04-12 84019316 7 0 1 N H H 0 , and Hd H

[127] In one aspect, provided is a pharmaceutical composition comprising a compound of any one of the preceding aspects and embodiments and a pharmaceutically acceptable excipient.

[128] In one aspect, provided is a method of inducing sedation and / or anesthesia in a subject, comprising administering to the subject an effective amount of a compound of the Fommla (I): HO\'' R2 (I), a pharmaceutically acceptable salt thereof, wherein: A is an optionally substituted nitrogencontaining heteroaryl or heterocyclyl; Lis -C(R3)(R~)-, -0-, -S-, or-NR3 -; R1 is hydrogen or C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, carbocyclyl, or heterocyclyl; R2 is hydrogen, C1-C6 alkyl (e.g., C1-C6 haloalkyl) or C1-C6 alkoxy; each R3 is independently hydrogen or C1-C6 alkyl; R5 is absent or hydrogen; and ::.::.:..::.:.: represents a single or double bond, wherein when one of ::.::.:..::.:.: is a double bond, the other ::.::.:..::.:.: is a single bond; and when one of the ::.::.:..::.:.: is a double bond, R5 is absent. 53 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[129] In one aspect, provided is a method of administering an effective amount of a compound, a pharmaceutically acceptable salt thereof, or pharmaceutical composition of a compound as described herein, e.g., a compound of the Formula (I), (Ia), (Ia-1), (Ia-2), (Ia-3), (Ia-4), (lb), (Ib-1), (Ib-2), (Ib-3), or (Ib-4), to a subject in need thereof, wherein the subject experiences sedation and / or anesthesia within two hours of administration.

[130] In some embodiments, the subject experiences sedation and / or anesthesia within one hour of administration.

[131] In some embodiments, the subject experiences sedation and / or anesthesia instantaneously.

[132] In some embodiments, the compound is administered by intravenous administration.

[133] In some embodiments, the compound is administered chronically. [134) In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[135] In some embodiments, the compound is administered in combination with another therapeutic agent. [136) In one aspect, provided is a method for treating seizure in a subject, comprising administering to the subject an effective amount of a compound of the Formula (1): HQ\l' R2 (I), a pharmaceutically acceptable salt thereof, wherein: A is an optionally substituted nitrogencontaining heteroaryl or heterocyclyl; Lis -C(R')(R3 )-, -0-, -S-, or -NR3 -; R1 is hydrogen or C 1-C6 alkyl, C 1-C6 alkenyl, C 1-C6 alkynyl, carbocyclyl, or heterocyclyl; R 2 is hydrogen, C 1-C6 alkyl (e.g., C 1-C6 haloalkyl) or C 1-C6 alkoxy; each R' is independently hydrogen or C 1-C6 alkyl; R5 is absent or hydrogen; and :.:..::.:.=.:: represents a single or double bond, wherein when one of :.:..::.:.=.:: is a double bond, the other :.:..::.:.=.:: is a single bond; and when one of the :.:..::.:.=.:: is a double bond, R5 is absent. 54 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[137] In one aspect, provided is a method for treating epilepsy or status or status epilepticus in a subject, the method comprising administering to the subject an effective amount of a compound of the Formula (I): HQ\l' R2 (I), a pham1aceutically acceptable salt thereof, wherein: A is an optionally substituted nitrogen containing heteroaryl or heterocyclyl; Lis -C(R')(R3 )-, -0-, -S-, or -NR3 -; R1 is hydrogen or C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, carbocyclyl, orheterocyclyl; R2 is hydrogen, C1-C6 alkyl (e.g., Ci-CG haloalkyl) or C1-C6 alkoxy; each R3 is independently hydrogen or C1-C6 alkyl; R5 is absent or hydrogen; and ::..:..::.:..:. represents a single or double bond, wherein when one of ::..:..::.:..:. is a double bond, the other ::..:..::.:..:. is a single bond; and when one of the ::..:..::.:..:. is a double bond, R5 is absent.

[138] In one aspect, provided is a method for treating disorders related to GABA function in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound, a pharmaceutically acceptable salt thereof, or pharmaceutical composition of one of a compound as described herein, e.g., a compound of the Formula (I), (Ia), (Ia-1), (Ia-2), (la-3), (Ia-4), (lb), (lb-1), (lb-2), (lb-3), or (lb-4).

[139] In one aspect, provided is a method for treating a CNS-related disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound as described herein, e.g., a compound of the Formula (I), (Ia), (Ia-1), (Ia-2), (Ia-3), (Ia-4), (lb), (lb-1 ), (Ib-2), (Ib-3), or (lb-4), or a pharmaceutically acceptable salt thereof. In some embodiments, the CNS-related disorder is a sleep disorder, a mood disorder, a schizophrenia spectmm disorder, a convulsive disorder, a disorder of memory and / or cognition, a movement disorder, a personality disorder, autism spectrum disorder, pain, traumatic brain injury, a vascular disease, a substance abuse disorder and / or withdrawal syndrome, or tinnitus. In some 55 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 embodiments, the subject is a subject with Rett syndrome, Fragile X syndrome, or Angelman syndrome. [140) In one aspect, provided is a kit comprising a solid composition comprising a compound as described herein, e.g., a compound of the Formula (I), (Ia), (Ia-1), (Ia-2), (Ia-3), (Ia-4), (lb), (lb-1), (Ib-2), (Ib-3), or (lb-4), and a sterile diluent. Pharmaceutical Compositions [141) In one aspect, the invention provides a pham1aceutical composition comprising a compound of the present invention (also referred to as the "active ingredient") and a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises an effective amount of the active ingredient. In certain embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the active ingredient. In certain embodiments, the pharmaceutical composition comprises a prophylactically effective amount of the active ingredient. [142) The pharmaceutical compositions provided herein can be administered by a variety of routes including, but not limited to, oral (enteral) administration, parenteral (by injection) administration, rectal administration, transdermal administration, intradermal administration, intrathecal administration, subcutaneous (SC) administration, intravenous (IV) administration, intramuscular (IM) administration, and intranasal administration. [143) Generally, the compounds provided herein are administered in an effective amount. The amount of the compmmd actually administered will typically be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like. [144) When used to prevent the onset of a CNS-disorder, the compounds provided herein will be administered to a subject at risk for developing the condition, typically on the advice and m1der the supervision of a physician, at the dosage levels described above. Subjects at risk for developing a particular condition generally include those that have a family history of the condition, or those who have been identified by genetic testing or screening to be particularly susceptible to developing the condition. 56 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[145] The pharmaceutical compositions provided herein can also be administered chronically ("chronic administration"). Chronic administration refers to administration of a compow1d or pharmaceutical composition thereof over an extended period oftime, e.g., for example, over 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc, or may be continued indefinitely, for example, for the rest of the subject's life. In certain embodiments, the chronic administration is intended to provide a constant level of the compound in the blood, e.g., within the therapeutic window over the extended period of time.

[146] The pharmaceutical compostions of the present invention may be further delivered using a variety of dosing methods. For example, in certain embodiments, the pharmaceutical composition may be given as a bolus, e.g., in order to raise the concentration of the compound in the blood to m1 effective level. The placement of the bolus dose depends on the systemic levels of the active ingredient desired throughout the body, e.g .. an intramuscular or subcutm1eous bolus dose allows a slow release of the active ingredient, while a bolus delivered directly to the veins (e.g., through an IV drip) allows a much faster delivery which quickly raises the concentration of the active ingredient in the blood to an effective level. In other embodiments, the pharmaceutical composition may be administered as a continuous infusion, e.g .. by IV drip, to provide maintenm1ce of a steady-state concentration of the active ingredient in the subject's body. Furthermore, in still yet other embodiments, the pharmaceutical composition may be administered as first as a bolus dose, followed by continuous infusion.

[147] The compositions for oral administration can take the form of bulk liquid solutions or suspensions, or bulk powders. More commonly, however, the compositions are presented in unit dosage forms to facilitate accurate dosing. The term "unit dosage forms" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, premeasured ampules or syringes of the liquid compositions or pills, tablets, capsules or the like in the case of solid compositions. In such compositions, the compound is usually a minor component (from about 0.1 to about 50% by weight or preferably from about 1 to about 40% by weight) with the remainder being various vehicles or excipients and processing aids helpful for forming the desired dosing form. 57 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[148] With oral dosing, one to five and especially two to four and typically three oral doses per day are representative regimens. Using these dosing patterns, each dose provides from about 0. 0 l to about 20 mg / kg of the compound provided herein, with preferred doses each providing from about 0.1 to about 10 mg / kg, and especially about 1 to about 5 mg / kg.

[149] Transdermal doses are generally selected to provide similar or lower blood levels than are achieved using injection doses, generally in an amount ranging from about 0.01 to about 20% by weight, preferably from about 0.1 to about 20% by weight, preferably from about 0.1 to about 10% by weight, and more preferably from about 0.5 to about 15% by weight.

[150] Injection dose levels range from about 0.1 mg / kg / hour to at least 20 mg / kg / hour, all for from about 1 to about 120 hours and especially 24 to 96 hours. A preloading bolus of from about 0.1 mg / kg to about 10 mg / kg or more may also be administered to achieve adequate steady state levels. The maximum total dose is not expected to exceed about 5 g / day for a 40 to 80 kg human patient.

[151] Liquid forms suitable for oral administration may include a suitable aqueous or nonaqueous vehicle with buffers, suspending and dispensing agents, colorants, flavors and the like. Solid forms may include, for example, any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

[152] Injectable compositions are typically based upon injectable sterile saline or phosphatebuffered saline or other injectable excipients known in the ait. As before, the active compound in such compositions is typically a minor component, often being from about 0.05 to 10% by weight with the remainder being the injectable excipient and the like.

[153] Transdermal compositions are typically formulated as a topical ointment or cream containing the active ingredient(s). When formulated as a ointment, the active ingredients will typically be combined with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredients may be formulated in a cream with, for example an oil-in-water cream base. Such transdermal formulations are well-known in the art and generally include additional 58 Date Rec;ue / Date Received 2024-04-12 84019316 ingredients to enhance the dermal penetration of stability of the active ingredients or Fonnulation. All such known transdennal formulations and ingredients are included within the scope provided herein.

[154] The compounds provided herein can also be administered by a transdermal device. Accordingly, transdermal administration can be accomplished using a patch either of the reservoir or porous membrane type, or of a solid matrix variety. [155) The above-described components for orally administrable, injectable or topically administrable compositions are merely representative. Other materials as well as processing techniques and the like are set forth in Part 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Company, Easton, Pennsylvania.

[156] The compounds of the present invention can also be administered in sustained release forms or from sustained release drng delivery systems. A description of representative sustained release materials can be found in Remington's Pharmaceutical Sciences. [ 157] The present invention also relates to the pharmaceutically acceptable acid addition salt of a compound of the present invention. The acid which may be used to prepare the pharmaceutically acceptable salt is that which fom1s a non-toxic acid addition salt, i.e., a salt containing pharmacologically acceptable anions such as the hydrochloride, hydroiodide, hydrobromide, nitrate, sulfate, bisulfate, phosphate, acetate, lactate, citrate, ta1trate, succinate, maleate, fumarate, benzoate, para-toluenesulfonate, and the like.

[158] In another aspect, the invention provides a pharmaceutical composition comprising a compom1d of the present invention and a pharmaceutically acceptable excipient, e.g., a composition suitable for injection, such as for intravenous (N) administration.

[159] Pharmaceutically acceptable excipients include any and all diluents or other liquid vehicles, dispersion or suspension aids, surface active agents, isotonic agents, preservatives, lubricants and the like, as suited to the particular dosage form desired, e.g., injection. General considerations in the formulation and / or manufacture of pharmaceutical compositions agents can be found, for example, in Remington's Pharmaceutical Sciences, Sixteenth Edition, E.W. Martin (Mack Publishing Co., Easton, Pa., 1980), and Remington: The Science and Practice of Pharmacy, 21 st Edition (Lippincott Williams & Will<lns, 2005). 59 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[160] For example, injectable preparations, such as sterile injectable aqueous suspensions, can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. Exemplary excipients that can be employed include, but are not limited to, water, sterile saline or phosphate-buffered saline, or Ringer's solution.

[161] In certain embodiments, the pharmaceutical composition further comprises a cyclodextrin derivative. The most common cyclodextrins are a-, ()- and y- cyclodextrins consisting of 6, 7 and 8 a-1 ,4-linked glucose units, respectively, optionally comprising one or more substituents on the linked sugar moieties, which include, but are not limited to, substituted or unsubstituted methylated, hydroxyalkylated, acylated, and sulfoalkylether substitution. In certain embodiments, the cyclodextrin is a sulfoalkyl ether ~-cyclodextrin, e.g., for example, sulfobutyl ether ~-cyclodextrin, also known as Captisol®. See, e.g., U.S. 5,376,645. In certain embodiments, the composition comprises hexapropyl-[3-cyclodextrin. In a more particular embodiment, the composition comprises hexapropyl-[3-cyclodextrin (10-50% in water).

[162] The injectable composition can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[163] Generally, the compounds provided herein are administered in an effective amount. The amount of the compound actually administered will typically be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the ach1al compound administered, the age, weight, response of the individual patient, the severity of the patient's symptoms, and the like.

[164] The compositions are presented in unit dosage forms to facilitate accurate dosing. The term "unit dosage forms" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetem1ined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include pre-filled, pre-measured ampules or syringes of the liquid compositions. In such compositions, the compound is usually a minor component (from about 0.1 % to about 50% by weight or preferably from about 1 % to about 60 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 40% by weight) with the remainder being various vehicles or carriers and processing aids helpful for forming the desired dosing fom1. [165) The compounds provided herein can be administered as the sole active agent, or they can be administered in combination with other active agents. In one aspect, the present invention provides a combination of a compound of the present invention and another pham1acologically active agent. Administration in combination can proceed by any technique apparent to those of skill in the art including, for example, separate, sequential, concurrent, and alternating administration.

[166] Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pham1acologist can design and / or perform such modification with ordinary experimentation. General considerations in the formulation and / or manufacture of pharmaceutical compositions can be found, for example, in Remington: The Science and Practice of Pharmacy 21 st ed., Lippincott Williams & Wilkins, 2005. Method'J (f l!'Je and Treatment [167) As generally described herein, the present invention is directed to C21-substituted neuroactive steroids designed, for example, to act as GABA modulators. In certain embodiments, such compounds are envisioned to be useful as therapeutic agents for the inducement of anesthesia and / or sedation in a subject. In some embodiments, such compounds are envisioned to be useful as therapeutic agents for treating a CNS-related disorder (e.g., sleep disorder, a mood disorder, a schizophrenia spectrum disorder, a convulsive disorder, a disorder of memory and / or cognition, a movement disorder, a personality disorder, autism spectrum disorder, pain, traumatic brain injury, a vascular disease, a substance abuse disorder and / or 61 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 withdrawal syndrome, or tinnitus) in a subject in need (e.g., a subject with Rett syndrome, Fragile X syndrome, or Angelman syndrome). [168) Thus, in one aspect, the present invention provides a method of inducing sedation and / or anesthesia in a subject, comprising administering to the subject an effective amount of a compound of the present invention or a composition thereof. In certain embodiments, the compound is administered by intravenous administration. [169) Earlier studies (see, e.g., Gee et al., European Joumal of Pharmacology, 136:419-423 (1987)) demonstrated that certain 3a-hydroxylated steroids are orders of magnitude more potent as modulators of the GABA receptor complex (GRC) than others had reported (see, e.g., Majewska eta!., Science 232:1004-1007 (1986); Harrison et al., J Pharmacol. Erp. Ther. 241 :346-353 ( 1987)). Majewska et al. and Harrison et al. taught that 3a-hydroxylated-5- reduced steroids are only capable of much lower levels of effectiveness. In vitro and in vivo experimental data have now demonstrated that the high potency of these steroids allows them to be therapeutically useful in the modulation of brain excitability via the GRC (see, e.g., Gee et al., European Journal of Pharmacology, 136:419-423 ( 1987); Wieland et al., Psychopharmacology 118(1):65-71 ( 1995)).

[170] Various synthetic steroids have also been prepared as neuroactive steroids. See, for example, U.S. Patent 5,232,917, which discloses neuroactive steroid compounds useful in treating stress, anxiety, insomnia, seizure disorders, and mood disorders, that are amenable to GRC-active agents, such as depression, in a therapeutically beneficial manner. Furthermore, it has been previously demonstrated that these steroids interact at a unique site on the GRC which is distinct from other known sites of interaction ( e.g., barbiturates, benzodiazepines, and GABA) where therapeutically beneficial effects on stress, anxiety, sleep, mood disorders and seizure disorders have been previously elicited (see, e.g., Gee, K.W. and Yamamura, H.I., "Benzodiazepines and Barbittrrates: Drugs for the Treatment of Anxiety, Insomnia and Seizure Disorders," in Central Nervous System Disorders, Horvell, ed., Marcel-Dekker, New York (1985), pp. 123-147; Lloyd, K.G. and Morselli, P.L., "Psychopharmacology of GABAergic Drngs," in Psychopharmacology: The Third Generation of Progress, H.Y. Meltzer, ed., Raven Press, N.Y. (1987), pp. 183-195; and Gee et al., European Journal of Pharmacology, 136:419- 62 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 423 (1987). These compounds are desirable for their duration, potency, and oral activity (along with other forms of administration). [171) Compom1ds of the present invention, as described herein, are generally designed to modulate GABA function, and therefore to act as neuroactive steroids for the treatment and prevention of CNS-related conditions in a subject. Modulation, as used herein, refers to the inhibition or potentiation of GABA receptor function. Accordingly, the compounds and pharmaceutical compositions provided herein find use as therapeutics for preventing and / or treating CNS conditions in mammals including humans and non-human mammals. Thus, and as stated earlier, the present invention includes within its scope, and extends to, the recited methods of treatment, as well as to the compounds for such methods, and to the use of such compounds for the preparation of medicaments useful for such methods. [172) Exemplary CNS conditions related to GABA-modulation include, but are not limited to, sleep disorders [e.g., insomnia], mood disorders [e.g., depression, dysthymic disorder (e.g., mild depression), bipolar disorder (e.g., I and / or II), anxiety disorders (e.g., generalized anxiety disorder (GAD), social anxiety disorder), stress, post-traumatic stress disorder (PTSD), compulsive disorders (e.g., obsessive compulsive disorder (OCD))], schizophrenia spectrum disorders [e.g., schizophrenia, schizoaffective disorder], convulsive disorders [e.g., epilepsy (e.g., status epilepticus (SE)), seizures], disorders of memory and / or cognition [e.g., attention disorders (e.g., attention deficit hyperactivity disorder (ADHD)), dementia (e.g., Alzheimer's type dementia, Lewis body type dementia, vascular type dementia], movement disorders [e.g., Huntington's disease, Parkinson's disease], personality disorders [ e.g., anti-social personality disorder, obsessive compulsive personality disorder], autism spectmm disorders (ASD) [e.g., autism, monogenetic causes of autism such as synaptophathy's, e.g., Rett syndrome, Fragile X syndrome, Angelman syndrome], pain [e.g., neuropathic pain, injury related pain syndromes, acute pain, chronic pain], traumatic brain injury (TBI), vascular diseases [e.g., stroke, ischemia, vascular malformations], substance abuse disorders and / or withdrawal syndromes [e.g., addition to opiates, cocaine, and / or alcohol], and tinnitus. [173) In yet another aspect, provided is a combination of a compom1d of the present invention and another pharmacologically active agent. The compounds provided herein can be administered as the sole active agent or they can be administered in combination with other 63 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 agents. Administration in combination can proceed by any technique apparent to those of skill in the art including, for example, separate, sequential, concurrent and alternating administration. [174) In another aspect, provided is a method of treating or preventing brain excitability in a subject susceptible to or afflicted with a condition associated with brain excitability, comprising administering to the subject an effective amount of a compound of the present invention to the subject. [175) In yet another aspect, provided is a method of treating or preventing stress or anxiety in a subject, comprising administering to the subject in need of such treatment an effective amount of a compound of the present invention, or a composition thereof.

[176] In yet another aspect, provided is a method of alleviating or preventing seizure activity in a subject, comprising administering to the subject in need of such treatment an effective amount of a compound of the present invention.

[177] In yet another aspect, provided is a method of alleviating or preventing insomnia in a subject, comprising administering to the subject in need of such treatment an effective amount of a compound of the present invention, or a composition thereof.

[178] In yet another aspect, provided is a method of inducing sleep and maintaining substantially the level of REM sleep that is found in normal sleep, wherein substantial rebound insomnia is not induced, comprising administering an effective amount of a compound of the present invention. [179) In yet another aspect, provided is a method of alleviating or preventing PMS or PND in a subject, comprising administering to the subject in need of such treatment an effective amount of a compound of the present invention.

[180] In yet another aspect, provided is a method of treating or preventing mood disorders in a subject, comprising administering to the subject in need of such treatment an effective amount of a compound of the present invention. In certain embodiments the mood disorder is depression.

[181] In yet another aspect, provided is a method of inducing anesthesia in a subject, comprising administering to the subject an effective amount of a compound of the present invention. 64 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[182] In yet another aspect, provided is a method of cognition enhancement or treating memory disorder by administering to the subject a therapeutically effective amount of a compound of the present invention. In certain embodiments, the disorder is Alzheimer's disease. In certain embodiments, the disorder is Rett syndrome.

[183] In yet another aspect, provided is a method of treating attention disorders by administering to the subject a therapeutically effective amount of a compound of the present invention. In certain embodiments, the attention disorder is ADHD.

[184] In certain embodiments, the compound is administered to the subject chronically. In certain embodiments, the compound is administered to the subject orally, subcutaneously, intramuscularly, or intravenously. Anesthesia I Sedation

[185] Anesthesia is a pharmacologically induced and reversible state of amnesia, analgesia, loss of responsiveness, loss of skeletal muscle reflexes, decreased stress response, or all of these simultaneously. These effects can be obtained from a single drug which alone provides the correct combination of effects, or occasionally with a combination of drugs ( e.g., hypnotics, sedatives, paralytics, analgesics) to achieve very specific combinations of results. Anesthesia allows patients to undergo surgery and other procedures without the distress and pain they would otherwise experience.

[186] Sedation is the reduction of irritability or agitation by administration of a pharmacological agent, generally to facilitate a medical procedure or diagnostic procedure.

[187] Sedation and analgesia include a continuum of states of consciousness ranging from minimal sedation (anxiolysis) to general anesthesia.

[188] Minimal sedation is also known as anxiolysis. Minimal sedation is a dmg-induced state during which the patient responds normally to verbal commands. Cognitive function and coordination may be impaired. Ventilatory and cardiovascular functions are typically wiaffected.

[189] Moderate sedation / analgesia (conscious sedation) is a drug-induced depression of consciousness during which the patient responds purposefully to verbal command, either alone or accompanied by light tactile stimulation. No interventions are usually necessary to maintain 65 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 a patent airway. Spontaneous ventilation is typically adequate. Cardiovascular function is usually maintained. [190) Deep sedation / analgesia is a drug-induced depression of consciousness during which the patient cannot be easily aroused, but responds purposefully (not a reflex \vithdrawal from a painful stimulus) following repeated or painful stimulation. Independent ventilatory function may be impaired and the patient may require assistance to maintain a patent airway. Spontaneous ventilation may be inadequate. Cardiovascular fi.mction is usually maintained.

[191] General anesthesia is a drug-induced loss of consciousness during which the patient is not arousable, even to painful stimuli. The ability to maintain independent ventilatory function is often impaired m1d assistance is often required to maintain a patent airway. Positive pressure ventilation may be required due to depressed spontaneous ventilation or drug-induced depression of neuromuscular function. Cardiovascular function may be impaired. [192) Sedation in the intensive care unit (ICU) allows the depression of patients' awareness of the environment and reduction of their response to external stimulation. It cm1 play a role in the care of the critically ill patient, and encompasses a wide spectnm1 of symptom control that will vary between patients, and among individuals throughout the course of their illnesses. Heavy sedation in critical care has been used to facilitate endotracheal tube tolerance and ventilator synchronization, often with neuromuscular blocking agents. [193) In some embodiments, sedation (e.g., long-term sedation, continuous sedation) is induced and maintained in the ICU for a prolonged period of time (e.g., I day, 2 days, 3 days, 5 days, 1 week, 2 week, 3 weeks, l month, 2 months). Long-term sedation agents may have long duration of action. Sedation agents in the ICU may have short elimination half-life. [194) Procedural sedation and analgesia, also referred to as conscious sedation, is a technique of administering sedatives or dissociative agents with or without analgesics to induce a state that allows a subject to tolerate unpleasant procedures while maintaining cardiorespiratory fimction. Anxiety Disorders 66 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[195] Anxiety disorder is a blanket tem1 covering several different forms of abnormal and pathological fear and anxiety. Current psychiatric diagnostic criteria recognize a wide variety of anxiety disorders. (196] Generalized anxiety disorder is a common chronic disorder characterized by long-lasting anxiety that is not focused on any one object or situation. Those suffering from generalized anxiety experience non-specific persistent fear and worry and become overly concerned with everyday matters. Generalized anxiety disorder is the most common anxiety disorder to affect older adults. (197] In panic disorder, a person suffers from brief attacks of intense terror and apprehension, often marked by trembling, shaking, confusion, dizziness, nausea, difficulty breathing. These panic attacks, defined by the AP A as fear or discomfort that abruptly arises and peaks in less than ten minutes, can last for several hours and can be triggered by stress, fear, or even exercise; although the specific cause is not always apparent. In addition to recurrent unexpected panic attacks, a diagnosis of panic disorder also requires that said attacks have chronic consequences: either worry over the attacks' potential implications, persistent fear of future attacks, or significant changes in behavior related to the attacks. Accordingly, those suffering from panic disorder experience symptoms even outside of specific panic episodes. Often, normal changes in heartbeat are noticed by a panic sufferer, leading them to think something is wrong with their heart or they are about to have another panic attack. In some cases, a heightened awareness (hypervigilance) of body functioning occurs during panic attacks, wherein any perceived physiological change is interpreted as a possible life threatening illness (i.e. extreme hypochondriasis).

[198] Obsessive compulsive disorder is a type of anxiety disorder primarily characterized by repetitive obsessions ( distressing, persistent, and intrusive thoughts or images) and compulsions (urges to perfonn specific acts or rituals). The OCD thought pattern may be likened to superstitions insofar as it involves a belief in a causative relationship where, in reality, one does not exist. Often the process is entirely illogical; for example, the compulsion of walking in a certain pattern may be employed to alleviate the obsession of impending harm. And in many cases, the compulsion is entirely inexplicable, simply an urge to complete a ritual triggered by 67 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 nervousness. In a minority of cases, sufferers of OCD may only experience obsessions, with no overt compulsions; a much smaller number of sufferers experience only compulsions. [199) The single largest category of anxiety disorders is that of Phobia, which includes all cases in which fear and anxiety is triggered by a specific stimulus or situation. Sufferers typically anticipate terrifying consequences from encountering the object of their fear, which can be anything from an animal to a location to a bodily fluid. [200) Post-tratm1atic stress disorder or PTSD is an anxiety disorder which results from a traumatic experience. Post-traumatic stress can result from an extreme situation, such as combat, rape, hostage situations, or even serious accident. It can also result from long term (chronic) exposure to a severe stressor, for example soldiers who endure individual battles but cannot cope with continuous combat. Common symptoms include flashbacks, avoidant behaviors, and depression. Neurodegenerative Diseases and Disorders

[201] The term "neurodegenerative disease" includes diseases and disorders that are associated with the progressive loss of structure or function of neurons, or death of neurons. Neurodegenerative diseases and disorders include, but are not limited to, Alzheimer's disease (including the associated symptoms of mild, moderate, or severe cognitive impairment); amyotrophic lateral sclerosis (ALS); anoxic and ischemic injuries; ataxia and convulsion ( including for the treatment and prevention and prevention of seizures that are caused by schizoaffective disorder or by drugs used to treat schizophrenia); benign forgetfulness; brain edema; cerebellar ataxia including McLeod neuroacanthocytosis syndrome (MLS); closed head injury; coma; contusive injuries (e.g., spinal cord injury and head injury); dementias including multi-infarct dementia and senile dementia; disturbances of consciousness; Down syndrome; drug-induced or medication-induced Parkinsonism (such as neuroleptic-induced acute akathisia, acute dystonia, Parkinsonism, or tardive dyskinesia, neuroleptic malignant syndrome, or medication-induced postural tremor); epilepsy; fragile X syndrome; Gilles de la Tourette's syndrome; head trauma; hearing impairment and loss; Huntington's disease; Lennox syndrome; levodopa-induced dyskinesia; mental retardation; movement disorders including akinesias and akinetic (rigid) syndromes (including basal ganglia calcification, corticobasal degeneration, 68 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 multiple system atrophy, Parkinsonism-ALS dementia complex, Parkinson's disease, postencephalitic parkinsonism, and progressively supranuclear palsy); muscular spasms and disorders associated with muscular spasticity or weakness including chorea (such as benign hereditary chorea, drug-induced chorea, hemiballism, Huntington's disease, neuroacanthocytosis, Sydenham's chorea, and symptomatic chorea), dyskinesia (including tics such as complex tics, simple tics, and symptomatic tics), myoclonus (including generalized myoclonus and focal cyloclonus), tremor (such as rest tremor, postural tremor, and intention tremor) and dystonia (including axial dystonia, dystonic writer's cramp, hemiplegic dystonia, paroxysmal dystonia, and focal dystonia such as blepharospasm, oromandibular dystonia, and spasmodic dysphonia and torticollis); neuronal damage including ocular damage, retinopathy or macular degeneration of the eye; neurotoxic injury which follows cerebral stroke, thromboembolic stroke, hemorrhagic stroke, cerebral ischemia, cerebral vasospasm, hypoglycemia, amnesia, hypoxia, anoxia, perinatal asphyxia and cardiac arrest; Parkinson's disease; seizure; status epilecticus; stroke; tinnitus; tubular sclerosis, and viral infection induced neurodegeneration (e.g., caused by acquired immunodeficiency syndrome (AIDS) and encephalopathies). Neurodegenerative diseases also include, but are not limited to, neurotoxic injury which follows cerebral stroke, thromboembolic stroke, hemorrhagic stroke, cerebral ischemia, cerebral vasospasm, hypoglycemia, amnesia, hypoxia, anoxia, perinatal asphyxia and cardiac arrest. Methods of treating or preventing a neurodegenerative disease also include treating or preventing loss of neuronal function characteristic of neurodegenerative disorder. Epilepsy

[202] Epilepsy is a brain disorder characterized by repeated seizures over time. Types of epilepsy can include, but are not limited to generalized epilepsy, e.g., childhood absence epilepsy, juvenile nyoclonic epilepsy, epilepsy with grand-mal seizures on awakening, West syndrome, Lennox-Gastaut syndrome, partial epilepsy, e.g., temporal lobe epilepsy, frontal lobe epilepsy, benign focal epilepsy of childhood. Status epilepticus (.';;;E) 69 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[203] Status epilepticus (SE) can include, e.g., convulsive status epilepticus, e.g., early status epilepticus, established status epilepticus, refractory status epilepticus, super-refractory status epilepticus; non-convulsive status epilepticus, e.g., generalized status epilepticus, complex partial status epilepticus; generalized periodic epileptiform discharges; and periodic lateralized epileptiform discharges. Convulsive status epilepticus is characterized by the presence of convulsive status epileptic seizures, and can include early status epilepticus, established status epilepticus, refractory status epilepticus, super-refractory status epilepticus. Early status epilepticus is treated with a first line therapy. Established stah1s epilepticus is characterized by status epileptic seizures which persist despite treatment with a first line therapy, and a second line therapy is administered. Refractory status epilepticus is characterized by status epileptic seizures which persist despite treatment with a first line and a second line therapy, and a general anesthetic is generally administered. Super refractory status epilepticus is characterized by status epileptic seizures which persist despite treatment with a first line therapy, a second line therapy, and a general anesthetic for 24 hours or more.

[204] Non-convulsive stah1s epilepticus can include, e.g., focal non-convulsive status epilepticus, e.g., complex partial non-convulsive status epilepticus, simple partial nonconvulsive status epilepticus, subtle non-convulsive status epilepticus; generalized nonconvulsive status epilepticus, e.g., late onset absence non-convulsive status epilepticus, atypical absence non-convulsive stah1s epilepticus, or typical absence non-convulsive status epilepticus.

[205] Compositions described herein can also be administered as a prophylactic to a subject having a CNS disorder e.g., a traumatic brain injury, status epilepticus, e.g., convulsive status epilepticus, e.g., early status epilepticus, established status epilepticus, refractory status epilepticus, super-refractory status epilepticus; non-convulsive status epilepticus, e.g., generalized status epilepticus, complex partial status epilepticus; generalized periodic epileptiform discharges; and periodic lateralized epileptiform discharges; prior to the onset of a seizure. Seizure

[206] A seizure is the physical findings or changes in behavior that occur after an episode of abnormal electrical activity in the brain. The term "seizure" is often used interchangeably with 70 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 "convulsion." Convulsions are when a person's body shakes rapidly and uncontrollably. During convulsions, the person's muscles contract and relax repeatedly. [207) Based on the type of behavior and brain activity, seizures are divided into two broad categories: generalized and partial ( also called local or focal). Classifying the type of seizure helps doctors diagnose whether or not a patient has epilepsy. [208) Generalized seizures are produced by electrical impulses from throughout the entire brain, whereas partial seizures are produced (at least initially) by electrical impulses in a relatively small part of the brain. The part of the brain generating the seizures is sometimes called the focus.

[209] There are six types of generalized seizures. The most common and dramatic, and therefore the most well known, is the generalized convulsion, also called the grand-mat seizure. In this type of seizure, the patient loses consciousness and usually collapses. The loss of consciousness is followed by generalized body stiffening (called the "tonic" phase of the seizure) for 30 to 60 seconds, then by violent jerking (the "clonic" phase) for 30 to 60 seconds, after which the patient goes into a deep sleep (the "postictal" or after-seizure phase). During grand-mal seizures, injuries and accidents may occur, such as tongue biting and urinary incontinence.

[210] Absence seizures cause a short loss of consciousness Gust a few seconds) with few or no symptoms. The patient, most often a child, typically intem1pts an activity and stares blankly. These seizures begin and end abruptly and may occur several times a day. Patients are usually not aware that they are having a seizure, except that they may be aware of "losing time."

[211] Myoclonic seizures consist of sporadic jerks, usually on both sides of the body. Patients sometimes describe the jerks as brief electrical shocks. When violent, these seizures may result in dropping or involuntarily throwing objects. [212) Clonic seizures are repetitive, rhythmic jerks that involve both sides of the body at the same time.

[213] Tonic seizures are characterized by stiffening of the muscles.

[214] Atonic seizures consist of a sudden and general loss of muscle tone, particularly in the arms and legs, which often results in a fall. 71 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[215] Seizures described herein can include epileptic seizures; acute repetitive seizures; cluster seizures; continuous seizures; unremitting seizures; prolonged seizures; recurrent seizures; status epilepticus seizures, e.g., refractory convulsive status epilepticus, non-convulsive status epilepticus seizures; refractory seizures; myoclonic seizures; tonic seizures; tonic-clonic seizures; simple partial seizures; complex partial seizures; secondarily generalized seizures; atypical absence seizures; absence seizures; atonic seizures; benign Rolandic seizures; febrile seizures; emotional seizures; focal seizures; gelastic seizures; generalized onset seizures; infantile spasms; Jacksonian seizures; massive bilateral myoclonus seizures; multifocal seizures; neonatal onset seizures; nocturnal seizures; occipital lobe seizures; post traumatic seizures; subtle seizures; Sylvan seizures; visual reflex seizures; or withdrawal seizures. Equivalents and Scope [216) In the claims articles such as "a," "an," and "the" may mean one or more than one unless indicated to the contrary or otherwise evident from the context. Claims or descriptions that include "or" between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The invention includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The invention includes embodiments in which more than one, or all of the group members are present in, employed in, or othenvise relevant to a given product or process.

[217] Furthermore, the invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive tem1s from one or more of the listed claims is introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. It should it be understood that, in general, where the invention, or aspects of the invention, is / are referred to as comprising particular elements and / or features, certain embodiments of the invention or aspects of the invention consist, or consist essentially 72 Date Rec;ue / Date Received 2024-04-12 84019316 of, such elements and / or features. For purposes of simplicity, those embodiments have not been specifically set forth in haec verba herein. It is also noted that the terms "comprising" and "containing" are intended to be open and permits the inclusion of additional elements or steps. Where ranges are given, endpoints are included. Furthermore, unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or sub-range within the stated ranges in different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

[218] This application refers to various issued patents, published patent applications, journal articles, and other publications. If there is a conflict between any of the incorporated references and the instant specification, the specification shall control. In addition, any particular embodiment of the present invention that falls within the prior art may be explicitly excluded from any one or more of the claims. Because such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the invention can be excluded from any claim, for any reason, whether or not related to the existence of prior art.

[219] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above Description. Those of ordinary skill in the art will appreciate that various changes and modifications to this description may be made without departing from the spirit or scope of the present invention. Examples

[220] In order that the invention described herein may be more fully understood, the following examples are set forth. The synthetic and biological examples described in this application are offered to illustrate the compounds, phannaceutical compositions and methods provided herein and are not to be construed in any way as limiting their scope. 73 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Afaterials and Methods

[221] The compounds provided herein can be prepared from readily available starting materials using the following general methods and procedures. It will be appreciated that where typical or preferred process conditions (i.e., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvent used, but such conditions can be determined by one skilled in the art by routine optimization.

[222] Additionally, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. The choice of a suitable protecting group for a particular functional group as well as suitable conditions for protection and deprotection are well known in the art. For example, nwnerous protecting groups, and their introduction and removal, are described in T. W. Greene and P. G. M. Wuts, Protecting Groups in Organic S)mthesis, Second Edition, Wiley, New York, 1991, and references cited therein.

[223] The compounds provided herein may be isolated and purified by known standard procedures. Such procedures include (but are not limited to) recrystallization, column chromatography, HPLC, or supercritical fluid chromatography (SFC). The following schemes are presented with details as to the preparation of representative heteroaryls and heterocyclyls that have been listed herein. The compounds provided herein may be prepared from known or commercially available starting materials and reagents by one skilled in the art of organic synthesis. Exemplary chiral columns available for use in the separation / purification of the enantiomers / diastereomers provided herein include, but are not limited to, CHIRALPAK® ADI 0, CHIRALCEL® OB, CHIRALCEL® OB-H, CHIRALCEL® OD, CHIRALCEL® OD-H, CHIRALCEL® OF, CHIRALCEL® OG, CHIRALCEL® OJ and CHIRALCEL® OK.

[224] 1H-NMR reported herein (e.g., for intermediates) may be a partial representation of the full NMR spectrum of a compound, e.g., a compound described herein. For example, the reported 1H NMR may exclude the region between o (ppm) of about 1 to about 2.5 ppm.

[225] Exemplary general method for preparative HPLC: Column: Waters RBridge prep 10 LJm Cl 8, 19*250 mm. Mobile phase: aectonitrile, water (NH4HCO3) (30 L water, 24 g NH4HCO3, 30 mL NH3.H2O). Flow rate: 25 mL / min 74 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417

[226] Exemplary general method for analytical HPLC: Mobile phase: A: water (IO mM NH4HCO3), B: acetonitrileGradient: 5%-95% B in 1.6 or 2 min Flow rate: 1.8 or 2 mL / min; Column: XBridge Cl 8, 4.6*50mm, 3.5 μmat 45 C. Synthetic Method~ Example 1. General Procedure A: Preparation of A / B-trans scaffolds 0 ~ O A1 KOH, (MeO)2SO2 DME, rt Meo1 ~ 0~ A2 CH(OEth, pTsOH 1,4-dioxane / EtOH, rt Meo1 ~ EtO~ A3 1. 1 atm H2, Pd / C EtOAc, rt 2. 2N HCI, rt OH 2±fW HO H A6 Meo1 ~ o~~~ H A4 NaH, Me3S01 DMSO,rt PCC, CH2Cl2 o •eta rt 0 Zt!W HO H A7 0 ~ o A EtPPh3Br, tBuOK THF, 80°C A5 1. BH3•THF, THF 0°Cto rt 2. aq. NaOH H202, rt ~ HO A A9 PCC, CH2Cl2 o •c to rt Meo----._ ~ HBr, Br2 ~ • MeOH, rt Ho" ~ A10 LiAIH4 , THF o •c to rt Step 1. Preparation of compound A2. Finely-ground potassium hydroxide (28.0 g, 165 mmol) was added to a solution of commercially available 19-hydroxyandrost-4-ene-3,17-dione (Al, 50.0 g, 165 mmol) in anhydrous 1,2-dimethoxyethane (500 mL) at 0 °C under nitrogen, after which methyl sulfate (43.7 g, 208 mmol) was added portionwise. The mixture was slowly warmed to room temperature, stirring for a total of 18 h, at which point TLC analysis of the mixture (7:3 hexanes / ethyl acetate) indicated completion of the reaction. The mixture was diluted with water (500 mL) and extracted with ethyl acetate (3 x 200 mL). The combined organic extracts were washed with saturated aqueous sodium chloride solution ( 100 mL ), dried 75 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 with anhydrous sodium sulfate and filtered. The solvents were removed under reduced pressure and the residue was purified by column chromatography on silica gel, eluting with heptane / ethyl acetate (2:1 ), to provide A2 as a yellow solid (26.8 g, 50%). Step 2. Preparation of compound A3. Triethyl orthoformate (6.2 mL, 37 mmol) and ptoluenesulfonic acid (400 mg, 9.3 mmol) were added to a solution of compound A2 (9.9 g, 31.0 mmol) in anhydrous 1,4-dioxane (40 mL) and anhydrous ethanol (30 mL) at room temperature under nitrogen, and the mixture was stirred for 1.5 h, at which point TLC analysis of the mixture (7:3 hexanes / ethyl acetate) indicated completion of the reaction. The mixture was diluted with saturated aqueous sodium bicarbonate solution (100 mL), poured into water (300 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic extract solvents were removed under reduced pressure and the residue was purified by colmnn chromatography on silica gel, eluting with heptane / ethyl acetate (2: 1), to provide compound A3 as a white solid (7.0 g, 66%). Step 3. Preparation of compound A4. A mixture of compound A3 (7.0 g, 20.3) and palladium on carbon (3.0 g, 10 wt. %) in anhydrous ethyl acetate (200 mL) was shaken under an atmosphere of hydrogen ( 1 atmosphere) at room temperature for 1 h, at which point TLC analysis of the mixture (2: 1 hexanes / ethyl acetate) indicated completion of the reaction. The atmosphere was exchanged for nitrogen and the mixture was filtered through a pad of Celite under reduced pressure, washing the filter cake with ethyl acetate (50 mL). The filtrate solvents were treated with 10% aqueous hydrochloric acid solution (100 mL) and the biphasic mixture was stirred for 30 min. The mixture was extracted with ethyl acetate (2 x 100 mL) and the combined organic extracts were washed sequentially with saturated aqueous sodium bicarbonate and saturated aqueous sodimn chloride solutions (50 mL each), dried with anhydrous sodium sulfate and filtered. The solvents were removed under reduced pressure and the residue was purified by column chromatography on silica gel, eluting with heptane / ethyl acetate ( 4: 1 ), to provide compound A4 as a colorless oil (3.9 g, 60%). Step 4. Preparation of compound AS. Sodium hydride (1.7 g, 45 mmol, 60% in mineral oil) was added portionwise to a solution of trimethylsulfoxonium iodide (9.1 g, 45 mmol) in 76 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 anhydrous dimethyl sulfoxide ( l 00 mL) at room temperature under nitrogen, and the mixture was stirred for 1 h, after which a solution of compound A4 (9.5 g, 29.8 mmol) in anhydrous dimethyl sulfoxide (100 mL) was added. The resulting mixture was stirred at room temperature for 12 h, at which point TLC analysis of the mixture (7:3 hexanes / ethyl acetate) indicated completion of the reaction. The m.ixture was diluted with water (500 mL) and extracted with methyl tert-butyl ether (2 x 300 mL). The combined organic extracts were washed with water (2 x 300 mL), dried with anhydrous magnesium sulfate and filtered. The solvents were removed under reduced pressure to provide compound AS as a colorless oil that was used in the next step without further purification (7.5 g, 76%). Step 5. Preparation of compound A6. Lithium aluminum hydride ( 67 mL, 67 mmol, 1 M solution in tetrahydrofuran) was added to a solution of crude compound AS (7.5 g, 22.2 mmol) in anhydrous tetrahydrofuran (5 mL) at 0 °C under nitrogen, after which the mixture was slowly warmed to room temperature, stirring for a total of 2 h, at which point TLC analysis of the mixture (7:3 hexanes / ethyl acetate) indicated completion of the reaction. The mixture was carefully treated with water (10 rnL) followed by saturated aqueous sodium chloride solution (30 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic extracts were dried with anhydrous magnesium sulfate, filtered and the solvents were removed m1der reduced pressure to provide compound A6 as a colorless oil that was used in the next step without further purification (5.5 g, 74%): LCMS mlz 319 [M+H-H2Ot. Step 6. Preparation of compound A7. Pyridinium chlorochromate (4.0 g, 19 11111101) was added in one portion to a solution of crude compound A6 (4.2 g, 12.5 mmol) in anhydrous dichloromethane (100 mL) at 0 °C under nitrogen. The mixture was slowly warmed to room temperature, stirring for a total of 3 h, at which point TLC analysis of the mixture (7:3 hexanes / ethyl acetate) indicated completion of the reaction. The solids were removed by filtration and the filtrate solvents were removed W1der reduced pressure. The residue was purified by column chromatography on silica gel, eluting with heptane / ethyl acetate (7:3), to provide compollild A7 as a light yellow solid (2.l g, 50%): LCMS m / z 317 [M+H-H2Of. 77 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Step 7. Preparation of compound AS. Potassium tert-butoxide (4.3 g, 38 mmol) was added to a mixture of ethyltriphenylphosphonium bromide (14.2 g, 38 mmol) in anhydrous tetrahydrofuran (30 mL) at room temperature under nitrogen, after which the mixture was heated to 80 °C and stirred for 1 h. A solution of compound A7 (3.1 g, 9.3 mmol) in anhydrous tetrahydrofuran ( 10 mL) was added, after which stirring at 80 °C was continued for 2 h, at which point TLC analysis of the mixture (7:3 hexanes / ethyl acetate) indicated completion of the reaction. The cooled mixture was diluted with water (30 mL) and saturated aqueous sodium chloride solution (20 mL) and extracted with ethyl acetate (2 x 100 mL). The combined organic extract solvents were removed under reduced pressure and the residue was purified by column chromatography on silica gel, eluting with heptane / ethyl acetate (7:3), to provide compound AS as an off-white solid (2.0 g, 66%): LCMS mlz 329 [M+H-H2Or. Step 8. Preparation of compound A9. Borane-tetrahydrofuran complex (20.0 mL, 20 mmol, 1 M solution in tetrahydrofuran) was added to a solution of compotmd AS (2.0 g, 5.8 mmol) in anhydrous tetrahydrofuran (15 mL) at 0 °C under nitrogen, after which the mixture was slowly warmed to room temperature, stirring for a total of 1 h. The mixture was cooled in an ice bath and 10% aqueous sodium hydroxide solution (12 mL) was slowly added, followed by 30% aqueous hydrogen peroxide solution (12 mL). The resulting mixture was warmed to room temperature and stirred for 1 h, at which point TLC analysis of the mixture (7:3 hexanes / ethyl acetate) indicated completion of the reaction. The mixture was extracted with dichloromethane (2 x 100 mL) and the combined organic extracts were washed with saturated aqueous sodium chloride solution (25 mL), dried with sodium sulfate and filtered. The solvents were removed under reduced pressure to provide crude compound A9 as a white solid that was used in the next step without further purification (2.5 g, >99%). Step 9. Preparation of compound AlO. Pyridinium chlorochromate (2.4 g, 11 mmol) was added in one portion to a solution of crude compound A9 (2.5 g, 6.9 mmol) in anhydrous dichloromethane (30 mL) at 0 °C under nitrogen. The mixture was slowly warmed to room temperature, stirring for a total of 2 h, at which point TLC analysis of the mixture (7:3 hexanes / ethyl acetate) indicated completion of the reaction. The solids were removed by 78 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 filtration and the filtrate solvents were removed under reduced pressure. The residue was purified by column chromatography on silica gel, eluting with heptane / ethyl acetate (7:3), to provide AlO as an off-white solid (1.5 g, 61%). Step 10. Preparation of compound All. Hydrogen bromide (3 drops, 48% in water) was added to a solution of AlO, 1.4 g, 3.9 mmol) in anhydrous methanol (150 mL) at room temperature in the dark under nitrogen, after which bromine (0.4 mL, 7.7 mmol) was added. The mixture was stirred for 1 h, at which point TLC analysis of the mixture (7:3 hexanes / ethyl acetate) indicated completion of the reaction. The mixture was poured into ice-water (100 mL), treated with saturated aqueous sodium bicarbonate solution (30 mL) and extracted with ethyl acetate (2 x 60 mL). The combined organic extracts were washed with saturated aqueous sodium bicarbonate solution (4 x 100 mL) and saturated aqueous sodium chloride solution (50 mL), dried with magnesium sulfate and filtered. The solvents were removed under reduced pressure and the residue was purified by column chromatography on silica gel, eluting with heptane / ethyl acetate (1:1), to provide compound All as a colorless semi-solid (1.2 g, 71%): LCMS mlz 441 [M+Ht. Example 2. General Procedure A: Preparation of A / B-trans scaffolds 0 ZtlW O A1 1. 1 atm H2, Pd / C EtOAc, rt 2. 2N HCI, rt KOH, (Et0)2S02 EtOl ~ DME, rt 0~ EtOl~ 0~ H A14 A12 NaH, Me3S0I DMSO, rt 79 Date Rec;ue / Date Received 2024-04-12 CH(OEt)a, pTsOH 1,4-dioxane / EtOH, rt 0 ~ o H A15 LiAIH4, THF 0 °C to rt WO 2015 / 027227 OH iW HO H A16 1. BH3•THF, THF 0°Ctort PCC, CH2Cl2 0 ·c to rt 0 itfW HO H A17 o•ctort EtPPh3Br, tBuOK THF, 80 °C PCT / US2014 / 052417 HBr, Br2 MeOH,rt 0 Step 1. Preparation of compound A12. Prepared according General Procedure A, Step 1 from Al, 10.0 g, 33 mmol) and ethyl sulfate ( 17.3 mL, 132 mmol), with purification by column chromatography on silica gel to provide compound A12 as a yellow oil (4.6 g, 42%). Step 2. Preparation of compound A13. Prepared according General Procedure A, Step 2 from compound A12 (4.6 g, 14 mmol) to provide crude compound A13 as a yellow oil that was used in the next step without further purification. Step 3. Preparation of compound A14. Prepared according General Procedure A, Step 3 from crude compound A13, with purification by column chromatography on silica gel to provide compound A14 as a yellow oil ( 1.5 g, 31 %). Step 4. Preparation of compound A15. Prepared according General Procedure A, Step 4 from compound A14 (1.7 g, 5.l mmol) to provide crude compound A15 as a yellow oil that was used in the next step without further purification. Step 5. Preparation of compound A16. Prepared according General Procedure A, Step 5 from crnde compound A15 to provide crude compound A16 as a yellow oil that was used in the next step without further purification. 80 Date Rec;ue / Date Received 2024-04-12 Br WO 2015 / 027227 PCT / US2014 / 052417 Step 6. Preparation of compound Al 7. Prepared according General Procedure A, Step 6 from crude compound Al6, with purification by column chromatography on silica gel to provide compound Al7 as an off-white solid (751 mg, 40%). Step 7. Preparation of compound Al 8. Prepared according General Procedure A, Step 7 from compound Al 7 (750 mg, 2.2 mmol), with purification by column chromatography on silica gel to provide compound A18 as a colorless oil (757 mg, 97%). Step 8. Preparation of compound Al 9. Prepared according General Procedure A, Step 8 from compom1d A18 (757 mg, 2.1 mmol), to provide crude compom1d A19 as a yellow oil that was used in the next step without further purification. Step 9. Preparation of A20. Prepared according General Procedure A, Step 9 from cmde compound A19, with purification by column chromatography on silica gel to provide A20 as a white solid (515 mg, 65%): mp 106-107 °C; 1HNMR (500 MHz, CDCb) 8 3.51 (d, J= 16.5 Hz, IH), 3.43-3.36 (m, 3H), 2.53 (t, J = 5.0 Hz, IH), 2.18-1.96 (m, 6H), 1.74-0.92 (m, 25H), 0.84- 0.82 (m, lH), 0.62 (s, 3H) ppm; ESI MS mlz 359 [M+H-H2Of. Step 10. Preparation of A21. Hydrogen bromide (10 drops, 48% in water) was added to a solution of A20 ( 490 mg, 1.30 mmol) in anhydrous methanol (40 mL) at room temperature in the dark under nitrogen, after which bromine (235 mg, 13.0 mmol) was added. The mixture was stirred for 1 h, at which point TLC analysis of the mixture (7:3 hexanes / ethyl acetate) indicated completion of the reaction. The mixture was poured into ice-water (100 mL), treated with saturated aqueous sodium bicarbonate solution (30 mL) and extracted with ethyl acetate (2 x 60 mL). The combined organic extracts were washed with saturated aqueous sodium bicarbonate solution (4 x 100 mL) and saturated aqueous sodium chloride solution (50 rnL), dried with magnesium sulfate and filtered. The solvents were removed under reduced pressure and the residue was purified by column chromatography on silica gel, eluting with heptane / ethyl acetate ( 1: 1 ), to provide compound A21 as a white solid ( 468 mg, 79% ). LCMS m / z 43 7 [M+H-H2Of 81 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Example 3. General Procedure B: Preparation of A / B-trans scaffold C-21 analogs 0 Br 5-chlorobenzotriazole 5-Chloro-1H-benzo[d][l,2,3]triazole (470 mg, 3.06 mmol) and potassium carbonate (704 mg, 5.1 mmol) were added to a solution of compound All (225 mg, 0.51 mmol) in anhydrous tetrahydrofuran (20 mL) at room temperature under nitrogen and the mixture was stirred for 16 h, at which point TLC analysis of the mixture (2: 1 hexanes / ethyl acetate) indicated completion of the reaction. The mixture was diluted with water (120 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic extracts were washed with saturated aqueous sodium chloride solution (60 mL), dried with sodium sulfate and filtered. The solvents were removed under reduced pressure and the residue was semi-purified by column chromatography on silica gel, eluting with hexanes / ethyl acetate (3:1), to provide a mixture of the three regioisomers. The residue was further purified by reverse phase preparative HPLC to provide 35 as an off-white solid (150 mg, 29%): mp 205-207 °C; 1H NMR (300 MHz, CDCb) 8 7.87 (dd, J = 1.8, 0.6 Hz, IH), 7.81 (dd, J = 9.0, 0.6 Hz, lH), 7.34 (dd, J = 9.0, 1.8 Hz, lH), 5.55 (d, .J,A.B = 17.1 Hz, lH), 5.46 (d, JAB= 17.1 Hz, lH), 3.48 (d, J = 9.9 Hz, IH), 3.38 (d, J = 10.2 Hz, lH), 3.30 (s, 3H), 2.66 (t, J = 8.7 Hz, lH), 2.30-2.18 (m, lH), 2.18-2.09 (m, lH), 2.09-2.00 (m, IH), 1.82-1.38 (m, llH), 1.38-1.06 (m, IOH), 1.06-0.92 (m, IH), 0.92-0.80 (m, lH), 0.76 (s, 3H) ppm; ESI MS mlz 514 [M+Ht. 82 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Further elution provided 36 as an off-white solid (86 mg, 17%): mp 97-101 °C; 1H NMR (300 MHz, CDC13) & 8.03-7.97 (m, lH), 7.36-7.31 (m, 2H), 5.42 (d, JAB= 18.3 Hz, lH), 5.34 (d, JAB = 18.0 Hz, lH), 3.49 (d, J = 9.9 Hz, IH), 3.38 (d, J = 9.9 Hz, lH), 3.31 (s, 3H), 2.72 (t, J = 8.7 Hz, IH), 2.30-2.10 (m, 2H), 2.10-2.00 (m, lH), 1.85-1.40 (m, 11H), 1.39-0.80 (m, 12H), 0.75 (s, 3H) ppm; ESI MS mlz 5140 [M+Hf. Further elution provided 37 as an off-white solid (112 mg, 21 %): mp 106-110 °C; 1H NMR (300 MHz, CDCh) & 8.06 (d, J= 1.2 Hz, lH), 7.45 (dd, J= 9.0, 1.5 Hz, lH), 7.27 (d, J= 8.7 Hz, lH), 5.41 (s, 2H), 3.49 (d, J = 9.9 Hz, IH), 3.38 (d, J = 10.2 Hz, lH), 3.30 (s, 3H), 2.71 (t, J = 8.7 Hz, IH), 2.29-2.00 (m, 3H), 1.83-1.44 (m, l lH), 1.44-0.82 (m, 12H), 0.74 (s, 3H) ppm; ESI MS mlz 514 [M+Hf". Example 4. Preparation of compound 8. Br 1-methylpiperazine K2C03, THF, rt Prepared according General Procedure B from compound All (50 mg, 0.114 mmol) and Nmethylpiperazine (227 mg, 2.27 mmol), with purification by reverse phase preparative HPLC to provide compound 8 as a white solid (36.6 mg, 70%): mp 136-137 °C; 1HNMR (500 MHz, CDCh) o 3.46 (d, J= 10.0 Hz, lH), 3.36 (d, J= 10.0 Hz, lH), 3.28 (s, 3H), 3.17 (s, 2H), 2.60- 2.51 (m, 8H), 2.30 (s, 3H), 2.18-2.14 (m, lH), 2.02 (dt, J = 13.0, 3.5 Hz, lH), 1.88 (dt, J = 12.0, 3.5 Hz, lH), 1.71-1.46 (m, lOH), 1.34-1.72 (m, l IH), 0.98-0.84 (m, lH), 0.84-0.80 (m, lH), 0.64 (s, 3H) ppm; ESI MS mlz 461 [M+Hf. Example 5. Preparation of compounds 3 and 1. 83 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Br triazole + Prepared according General Procedure B from compound All (300 mg, 0.67 mmol) and lH- 1,2,3-triazole (188 mg, 2.71 mmol), with purification by reverse phase preparative HPLC to provide compound 3 as a white solid (36.6 mg, 70%): mp 72-74 °C; 1HNMR (300 MHz, CDCb) 5 7.75 (d, J= 1.2 Hz, IH), 7.63 (d, J= 1.2 Hz, lH), 5.20 (q, J= 18.0 Hz, 2H), 3.46 (d, J= 9.9 Hz, lH), 3.37 (d, J = 10.2 Hz, lH), 2.65 (t, J = 9.0 Hz, lH), 2.09-2.01 (m, lH), 1.78-1.68 (m, 4H), 1.60-1.49 (m, 13H), 1.46-1.26 (m, 9H), 1.23-0.87 (m, 2H), 0.63 (s, 3H) ppm; ESI MS mlz 430 [M+Ht. Further elution provided compound 1 as an off-white solid (110 mg, 38%): mp 157-150 °C; 1HNMR 8 7.67 (s, 2H), 5.23 (q, J = 17.0 Hz, 2H), 3.47 (d, J = 10.0 Hz, lH), 3.37 (d, J = 10.0 Hz, lH), 3.28 (s, 3H), 2.57 (t, J= 9.5 Hz, lH), 2.24-2.17 (m, lH), 2.09-2.01 (m, 2H), 1.75-1.67 (m, 4H), 1.62-1.47 (rn, l0H), 1.38-1.23 (m, 3H), 1.23-1.08 (m, 4H), 1.02-0.92 (m, lH), 0.86-0.81 (m, lH), 0.73 (s, 3H) ppm; ESI MS mlz 430 [M+Ht. Example 6. Preparation of compound 13. Br 4-cyanopyrazole N-N y CN Prepared according General Procedure B from compound All (25 mg, 0.057 mmol) and 5- chlorotriazole (106 mg, 1.14 mrnol), with purification by reverse phase preparative HPLC to provide cornpow1d 13 as a white solid (16.8 mg, 65%): mp 141-142 °C; 1HNMR (500 MHz, CDCh) 8 7.85 (s, lH), 7.80 (s, lH), 4.95 (dd, J = 62.5, 17.5 Hz, 2H), 3.47 (dd, J = 10.0 Hz, lH), 3.37 (dd, J = 10.0 Hz, IH), 3.28 (s, 3H), 2.60 (t, J = 9.0 Hz, lH), 2.23-2.20 (m, lH), 2.05-2.01 84 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 (m, 2H), 1.76-1.69 (m, 4H), 1.63-1.49 (m, 7H), 1.47-1.10 (m, I0H), 0.99-0.97 (m, lH), 0.87-0.85 (m, IH), 0.69 (s, 3H) ppm; ESI MS mlz 436 [M+H-H2Of'. Example 7. Preparation of compounds 14 and 15. Br tetrazole Prepared according General Procedure B from compound All (60 mg, 0.14 mmol) and tetrazole (57 mg, 0.81 mmol), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide compound 15 as an off-white solid (6 mg, 10%): mp 88-91 °C; 1H NMR (500 MHz, CDCb) 8 8.73 (s, lH), 5.30 (d,JAB = 18.5 Hz, lH), 5.17 (d,JAB = 18.5 Hz, lH), 3.47 (d, J = 10.0 Hz, IH), 3.37 (d, J = 10.0 Hz, IH), 3.29 (s, 3H), 2.66 (t, J = 9.0 Hz, lH), 2.28-2.20 (m, IH), 2.07-2.00 (m, 2H), 1.82-1.69 (m, 4H), 1.65-1.40 (m, 7H), 1.35- 1.09 (m, lOH), 1.04-0.95 (m, IH), 0.92-0.83 (m, IH), 0.69 (s, 3H) ppm; ESI MS mlz 431 [M+Ht. Further elution provided compound 14 as an off-white solid (7 mg, 12%): mp 72-75 °C; 1H NMR (500 MHz, CDCh) 8 8.56 (s, lH), 5.47 (d, !till= 17.0 Hz, lH), 5.42 (d, !till= 17.5 Hz, IH), 3.47 (d, J = 10.5 Hz, IH), 3.38 (d, J = 10.0 Hz, IH), 3.29 (s, 3H), 2.64 (t, J = 9.0 Hz, lH), 2.27-2.19 (m, lH), 2.18-2.00 (m, 2H), 1.80-1.68 (m, 4H), 1.66-1.46 (m, 6H), 1.44-1.37 (m, IH), 1.35-1.08 (m, l0H), 1.04-0.94 (m, IH), 0.90-0.83 (m, lH), 0.74 (s, 3H) ppm; ESI MS mlz 431 [M+Ht. Example 8. Preparation of compounds 16 and 17. Br 5-Me-tetrazole Hcf H 17 85 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Prepared according General Procedure B from compound All (215 mg, 0.49 mmol) and 5- methyl-lH-tetrazole (253 mg, 2.92 mmol), with semi-purification by colmm1 chromatography on silica gel followed by reverse phase preparative HPLC to provide 16 as an off-white solid (56 mg, 26%): mp 88-91 °C; 1H NMR (300 MHz, CDC13) o 5.13 (d, J,A.B = 18.0 Hz, lH), 5.06 (d, J,A.B = 18.0 Hz, lH), 3.48 (d, J= 10.0 Hz, lH), 3.37 (d, J= 10.0 Hz, lH), 3.29 (s, 3H), 2.66 (t, J= 9.0 Hz, lH), 2.47 (s, 3H), 2.27-2.15 (m, lH), 2.08-1.98 (m, 2H), 1.85-1.38 (m, l lH), 1.37-0.95 (m, l lH), 0.91-0.83 (m, lH), 0.70 (s, 3H) ppm; ESI MS mlz 445 [M+HJ1. Further elution afforded 17 as an off-white solid (95 mg, 44%): mp 71-74 °C; 1H NMR (500 MHz, CDCh) o 5.37 (d, . / A.B = 17.0 Hz, lH), 5.32 (d, JAB= 17.5 Hz, lH), 3.47 (d, J = 10.0 Hz, lH), 3.37 (d, J = 10.0 Hz, IH), 3.29 (s, 3H), 2.62 (t, J = 9.0 Hz, lH), 2.56 (s, 3H), 2.26-2.18 (m, lH), 2.09-2.00 (m, 2H), 1.80-1.68 (m, 4H), 1.65-1.46 (m, 6H), 1.43-1.08 (m, llH), 1.04-0.94 (m, IH), 0.90-0.82 (m, lH), 0.73 (s, 3H) ppm; ESI MS mlz 445 [M+Ht. Example 9. Preparation of compound 18. 0 Br 4-Cl-1-pyrazole K2C03, THF, rt 18 Prepared according General Procedure B from compound All (21 mg, 0.047 mmol) and 4- chloro-IH-pyrazole (29 mg, 0.28 mmol), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 18 as an off-white solid ( 10 mg, 46%): mp 100-104 °C; 1H NMR (500 MHz, CDCb) o 7.45 (s, lH), 7.40 (s, lH), 4.90 (d, JAB = 17.5 Hz, lH),4.80 (d,J,A.B = 17.5 Hz, lH), 3.46 (d,J= 10.0Hz, lH), 3.37 (d,J= 10.0 Hz, lH), 3.28 (s, 3H), 3.13 (s, 3H), 2.57 (t, J= 9.0 Hz, IH), 2.26-2.16 (m, IH), 2.07-1.98 (m, 2H), l.76- 1.67 (m, 4H), 1.63-1.46 (m, 6H), 1.41-1.07 (m, llH), 1.03-0.92 (m, lH), 0.91-0.80 (m, lH), 0.69 (s, 3H) ppm; ESI MS mlz 463 [M+Ht. Example 10. Preparation of compound 19. 86 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 Br 3-CN-1-pyrazole K2C03, THF, rt PCT / US2014 / 052417 N--N 0--cN 19 Prepared according General Procedure B from compound All (30 mg, 0.06 mmol) and lHpyrazole- 3-carbonitrile (25 mg, 0.03 mmol), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 19 as an off-white solid ( 17 mg, 56%): mp 115-120 °C; 1HNMR (300 MHz, CDCh) o 7.47 (d, J= 2.4 Hz, IH), 6.72 (d, J= 2.4 Hz, lH), 5.00 (q, J= 18.0 Hz, 2H), 3.49 (d, J= 9.9 Hz, lH), 3.39 (d,J= 10.2 Hz, lH), 3.29 (s, 3H), 2.65 (t, J = 9.0 Hz, IH), 2.09-2.01 (m, IH), 1. 78-1.68 (m, 4H), 1.60-1.49 (m, IOH), 1.46- 1.26 (m, 9H), 1.23-0.87 (m, 2H), 0.63 (s, 3H) ppm; ESI MS mlz 436 [M+H-H20t. Example 11. Preparation of compound 20. 0 Br 4-Me-1-pyrazole A Prepared according General Procedure B from compound All (130 mg, 0.29 mmol) and 4- methyl-IH-pyrazole (247 mg, 3.01 mmol) with the substitution of cesitm1 carbonate (480 mg, 1.5 mmol) in anhydrous acetonitrile (8 mL), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 20 as an off-white solid (15 mg, 11 %): mp 67-71 °C; 1H NMR (500 MHz, CDCh) o 7.33 (s, IH), 7.16 (s, lH), 4.87 (d, JAB= 18.0 Hz, lH), 4.79 (d, JAB= 17.5 Hz, lH), 3.46 (d, J= 10.0 Hz, lH), 3.37 (d, J= 10.0 Hz, lH), 3.28 (s, 3H), 3.13 (s, 3H), 2.56 (t, J = 8.5 Hz, lH), 2.23-2.15 (m, lH), 2.09 (s, 3H), 2.07-2.00 (m, 2H), 1.75-1.65 (m, 4H), 1.62-1.45 (m, 6H), 1.40-1.08 (m, 11H), 1.02-0.94 (m, lH), 0.89- 0.82 (m, lH), 0.69 (s, 3H) ppm; ESI MS mlz 443 [M+Ht. Example 12. Preparation of compound 21. 87 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Br 2-F-imidazole Prepared according General Procedure B from compound All (50 mg, 0.13 mmol) and 2- fluoroimidazole hydrochloride (75 mg, 0.61 mmol) with the substitution of cesium carbonate (200 mg, 0.62 mmol) in anhydrous acetonitrile (4 mL), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 21 as a yellow solid (35 mg, 69%): mp 78-81 °C; 1H NMR (300 MHz, CDCh) 8 6.91 (d, J= 1.5 Hz, lH), 6.64 (s, J = 1.5 Hz, lH), 5.54 (d, JAB= 18.3 Hz, lH), 4.41 (d, J,'IB = 18.0 Hz, lH), 3.48 (d, J = 10.0 Hz, lH), 3.37 (d, J = 10.0 Hz, lH), 3.30 (s, 3H), 2.54 (t, J = 9.0 Hz, lH), 2.28-2.14 (m, IH), 2.09-1.98 (m, 2H), 1.80-1.64 (m, 4H), 1.64-1.46 (m, 6H), 1.44-0.80 (m, 13H), 0.69 (s, 3H) ppm; ESI MS m / z 470 [M+Ht. Example 13. Preparation of compound 23. Br 4-Me-imidazole Prepared according General Procedure B from compound All (75 mg, 0.17 mmol) and 4- methylimidazole (279 mg, 3.4 mmol), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 23 as a white solid (18 mg, 24%): mp 87-89 °C; 1H NMR (500 MHz, CDCh) 8 7.40 (s, lH), 6.56 (s, lH), 4.63 (dd, J= 18.0, 10.8 Hz, 2H), 3.47 (d, J = 10.0 Hz, lH), 3.37 (d, J = 10.0 Hz, lH), 3.29 (s, 3H), 2.60-2.54 (m, lH), 2.24-2.17 (m, 3H), 2.08 (s, IH), 2.04-2.01 (m, lH), 1.96-1.93 (m, lH), 1.76-1.68 (m, 4H), 1.63-1.46 (111, 6H), 1.42-1.09 (111, llH), 1.02-0.93 (m, IH), 0.87-0.82 (111, lH), 0.68 (s, 3H) ppm; APCI MS mlz 443 [M+Ht. 88 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 Example 14. Preparation of compound 24. Br 4-S02Me-pyrazole K2C03, THF, rt PCT / US2014 / 052417 Hd H 24 Prepared according General Procedure B from compound All (100 mg, 0.23 mmol) and 4- (methylsulfonyl)-lH-pyrazole (99 mg, 0.68 mmol), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 24 as a white solid (6 mg, 5%): mp 90-92 °C; 1H NMR (500 MHz, CDCh) o 7.91 (s, IH), 7.86 (s, IH), 5.02 (d, JAB= 18.0 Hz, lH), 4.90 (d, JAB= 17.5 Hz, IH), 3.47 (d, J = 10.0 Hz, lH), 3.37 (d, J = 10.0 Hz, IH), 3.28 (s, 3H), 3.13 (s, 3H), 2.61 (t, J= 9.0 Hz, lH), 2.26-2.16 (m, IH), 2.07-2.00 (m, 2H), 1.78-1.68 (m, 4H), 1.64-1.46 (m, 6H), 1.44-1.36 (111, lH), 1.35-1.08 (m, l0H), 1.04- 0.94 (m, lH), 0.89-0.82 (m, lH), 0.69 (s, 3H) ppm; APCI MS mlz 507 [M+Hf. Example 15. Preparation of compounds 25 and 26. 0 Br A21 5-methyltetrazole K2C03, THF, rt 0 + Prepared according General Procedure B from compound A21 (150 mg, 0.33 mmol) and 5- methyltetrazole (554 mg, 6.6 mmol), with purification by reverse phase preparative HPLC to provide 25 as a white solid (43.6 mg, 28%): mp 71-72 °C; 1HNMR (500 MHz, CDCh) o 5.34 (dd, J = 26.5, 17.5 Hz, 2H), 3.52 (d, J = 10.0 Hz, lH), 3.42-3.38 (m, 3H), 2.62 (t, J = 9.0 Hz, IH), 2.56 (s, 3H), 2.24-2.22 (m, lH), 2.08-2.03 (m, 2H), 1.75-1.48 (m, llH), 1.39-1.08 (m, 13H), 0.99-0.98 (m, lH), 0.86-0.85 (m, lH), 0.72 (s, 3H) ppm; ESI MS mlz 459 [M+Ht. Further elution provided 26 as a white solid (14.9 mg, 7%): mp 82-83 °C; 1HNMR (500 MHz, CDCb) o 5.09 (dd, J = 37.0, 18.0 Hz, 2H), 3.52 (d, J = 10.0 Hz, lH), 3.43-3.37 (m, 3H), 2.65 (t, J = 9.0 Hz, lH), 2.47 (s, 3H), 2.24-2.22 (m, lH), 2.06-2.03 (m, 2H), 1.75-1.41 (m, llH), 1.33- 89 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 1.09 (m, 13H), 0.99-0.98 (m, lH), 0.87-0.86 (m, IH), 0.69 (s, 3H) ppm; ESI MS mlz 459 [M+Hf. Example 16. General Procedure C: Preparation of A / B-cis scaffolds HO)~ 0~ A1 NaH, Mel 50 psi H2, Pd / C THF, rt 0) Me~l [;no 0 ODW H C2 THF, 30°C o--QErf" ~O H C4 2N HCI, rt THF / acetone 0 =afW MeO H ca 1. BH3•THF, THF o•ctort 2.aq. NaOH H2O2, rt 1. EtPPh3Br, tBuOK THF, 60°C 2. 2N HCI, rt Meo..._~ ~ O H C7 PCC,CH2Cl2 o·ctort HOCH2CH20H pyridine • HCI toluene, reflux 0 oafW H CS MeMgBr, MeAl(OAr)z toluene, -78 •c to rt HBr, Br2 MeOH,rt 12, MeOH 60°c ~ HO H CS Step 1. Preparation of compound C2. A mixture of commercially available 19- hydroxyandrost-4-ene-3, l 7-dione (Al, 13.6 g, 45 mmol) and palladium on carbon (3.2 g, 10 wt. % ) in anhydrous tetrahydrofuran ( 150 mL) was shaken under an atmosphere of hydrogen ( 50 psi) at room temperature for 12 h, at which point TLC analysis of the mixture (2:1 hexanes / ethyl acetate) indicated completion of the reaction. The atmosphere was exchanged for nitrogen and the mixture was filtered through a pad of Celite under reduced pressure, washing the filter cake with ethanol. The filtrate solvents were removed under reduced pressure to provide C2 as a white solid that was used in the next step without further purification (13.0 g, 95%): LCMS mlz 305 [M+Ht. 90 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Step 2. Preparation of compound C3. Pyridine hydrochloride (750 mg, 6.5 mmol) was added to a solution of crude compound C2 (15.0 g, 49 mmol) in ethylene glycol (65 mL) and anhydrous toluene (180 mL) at room temperature under nitrogen. The mixture was heated at reflux for 12 h with water removal by Dean-Stark apparatus, at which point TLC analysis of the mixture (2: 1 hexanes / ethyl acetate) indicated completion of the reaction. The solvents were removed from the cooled mixture under reduced pressure and the residue was treated with saturated aqueous sodium chloride solution (50 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic extracts were washed with saturated aqueous sodium chloride solution (3 x 10 mL), dried with anhydrous sodium sulfate and filtered. The solvents were removed under reduced pressure to provide compound C3 as a colorless oil that was used in the next step without further purification (20.3 g, >99%): 1H NMR (300 MHz, CDCh) o 4.11-3.81 (m, 8H), 3.60-3.54 (m, IH), 2.05-1.92 (m, 3H), 1.81-163 (m, 4H), 1.59-1.35 (m, 12H), 1.28-1.12 (m, 5H), 0.8 (s, 3H) ppm; LCMS mlz 393 [M+Hf. Step 3. Preparation of compound C4. A solution of crude compound C3 (20.3 g, 49 mmol) in anhydrous tetrahydrofuran (120 mL) was added dropwise to a suspension of sodium hydride (7.9 g, 197 mmol, 60% in mineral oil) in anhydrous tetrahydrofuran (120 mL) at 0 °C under nitrogen, after which the mixture was stirred at 0 °C for 30 min. Iodomethane (15.3 mL, 246 mmol) was added dropwise, after which the mixture was heated to 35 °C and stirred for 3 h, at which point TLC analysis of the mixture (3:1 hexanes / ethyl acetate) indicated completion of the reaction. The cooled mixture was treated with saturated ammonium chloride solution ( I 00 mL) and extracted with ethyl acetate (2 x 50 mL). The combined organic extracts were washed with saturated aqueous sodium chloride solution (2 x 20 mL), dried with anhydrous sodium sulfate and filtered. The solvents were removed under reduced pressure to provide crude compound C4 as a yellow oil that was used in the next step without further purification (25.6 g, >99%): LCMS mlz 407 [M+Hf. Step 4. Preparation of compound C5. A mixture of crude compound C4 (25.5 g, 49 mmol) in tetrahydrofuran (150 mL) and acetone (90 mL) at room temperature was treated with 2N HCl (123 mL) and the mixture was stirred for 16 h, at \vhich point TLC analysis of the mixture (2:1 hexanes / ethyl acetate) indicated completion of the reaction. The reaction mixture was adjusted to pH 8 with slow addition of saturated aqueous sodium bicarbonate solution and extracted with 91 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 ethyl acetate (3 x 125 mL). The combined organic extracts were washed with saturated aqueous sodium chloride solution (2 x 20 mL), dried with anhydrous sodium sulfate and filtered. The solvents were removed under reduced pressure and the residue was purified by column chromatography on silica gel, eluting with hexanes / ethyl acetate (2: 1 ), to provide compound CS as a white solid (10.6 g, 67%): 1H NMR (300 MHz, CDCh) 8 3.62-3.59 (m, lH), 3.36-3.33 (m, 4H), 2.67-2.63 (m, lH), 2.58-2.45 (m, lH), 2.42-2.27 (m, 3H), 2.25-1.84 (m, 6H), 1.71-1.23 (m, l lH), 0.89 (s, 3H) ppm; LCMS mlz 319 [M+Hf. Step 5. Preparation of compound C6. Iodine (84 mg, 0.3 mmol) was added to a solution of compound CS (10.6 g, 33 mmol) in anhydrous methanol (200 mL) at room temperature under nitrogen, after which the mixture was heated to 60 °C and stirred for 90 min, at which point TLC analysis of the mixture (2: 1 hexanes / ethyl acetate) indicated completion of the reaction. The cooled mixture was treated with lN sodium hydroxide solution (200 mL) and extracted with hexanes / ethyl acetate (3:1, 3 x 100 mL). The combined organic extracts were washed with saturated aqueous sodium chloride solution (2 x 25 mL), dried with anhydrous sodium sulfate and filtered. The solvents were removed under reduced pressure to provide compound C6 as a colorless oil that was used in the next step without further purification (13.8 g, >99%); LCMS mlz 365 [M+Hr. Step 6. Preparation of compound C7. Potassium tert-butoxide (11.2 g, 100 mmol) was added to a mixture of ethyltriphenylphosphonium bromide (36.9 g, 100 mmol) in anhydrous tetrahydrofuran ( 150 mL) at room temperature under nitrogen, after which the mixture was heated to 60 °C and stirred for 4 h. A solution of compound C6 (13.8 g, 33 mmol) in anhydrous tetrahydrofuran ( 100 mL) was added, after which stirring at 60 °C was continued for 18 h. The cooled mixture was diluted with water (200 mL) and hexanes (100 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic extracts were washed with saturated aqueous sodium chloride solution (2 x 25 mL), treated with 2N HCl (100 mL) and stirred at room temperature for 3 h. The resulting mixture was washed with saturated aqueous sodium bicarbonate and saturated aqueous sodium chloride solutions, dried with sodium sulfate and filtered. The solvents were removed under reduced pressure and the residue was purified by column chromatography on silica gel, eluting with hexanesiethyl acetate (9: l ), to provide compound C7 as a colorless oil (9.2 g, 84%): LCMS mlz 331 [M+Hf. 92 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Step 7. Preparation of compound CS. Bis(2,6-di-tert-butyl-4- methylphenoxide)methylaluminum (40.6 mL, 16 mmol, 0.4 Min toluene) was added in one portion to a solution of compound C7 (1.8 g, 5.4 mmol) in anhydrous toluene (20 mL) at -78 °C under nitrogen, after which the mixture was stirred for 10 min. Methylmagnesium bromide (11.6 mL, 16 mmol, 1.4 Min tetrahydrofuranitoluene) was added dropwise, after which the mixture was stirred at -78 °C for 1 h. The mixture was warmed to ice bath temperature and slowly treated with 2N HCl ( 60 mL), warmed to room temperature and extracted with ethyl acetate (3 x 50 mL). The combined organic extracts were washed with saturated aqueous sodium chloride solution (2 x 20 mL), dried with sodium sulfate and filtered. The solvents were removed under reduced pressure and the residue was purified by cohunn chromatography on silica gel, eluting with hexanes / ethyl acetate (2: 1 ), to provide crude compound CS as a white semi-solid ( 1.5 g, 91%); LCMS m / z 347 [M+Ht. Step 8. Preparation of compound C9. Borane-tetrahydrofuran complex (27.6 mL, 27.6 mmol, 1.0 M solution in tetrahydrofuran) was added to a solution of compound CS (2.4 g, 6.9 mmol) in anhydrous tetrahydrofuran (24 mL) at 0 °C under nitrogen, after which the mixture was slowly warmed to room temperature, stirring for a total of 4 h. The mixture was cooled in an ice bath and 10% aqueous sodium hydroxide solution (20 mL) was slowly added, followed by 30% aqueous hydrogen peroxide solution (20 mL). The resulting mixture was wam1ed to room temperature and stirred for 1 h and then treated with saturated aqueous sodium chloride solution (100 mL) and extracted with dichloromethane (3 x 100 mL). The combined organic extracts were washed with saturated aqueous sodium chloride solution (25 mL), dried with sodium sulfate and filtered. The solvents were removed under reduced pressure to provide crude compound C9 as a white solid that was used in the next step without further purification (2.7 g, >99%); LCMS m / z 365 [M+Ht. Step 9. Preparation of ClO. Pyridinium chlorochromate (6.0 g, 28 mmol) was added in one portion to a solution of compound C9 (2.7 g, 6.9 mmol) in dichloromethane (100 mL) at 0 °C under nitrogen, after which the mixture was slowly warmed to room temperature, stirring for a total of 16 h. The solids were removed by filtration and the filtrate solvents were removed under reduced pressure. The residue was semi-purified by column chromatography on silica gel, eluting with hexanes / ethyl acetate (1:1), followed by further purification by reverse phase 93 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 preparative HPLC to provide ClO as a white solid (2.15 g, 86%): mp 142-144 °C; 1HNMR (300 MHz, CDCh) 8 3.55 (d, J= 9.0 Hz, lH), 3.33 (s, 3H), 3.19 (d, J= 9.0 Hz, lH), 2.53 (t, J= 9.0 Hz, IH), 2.21-2.11 (111, 4H), 2.08-1.87 (m, 3H), 2.14-1.91 (m, 7H), 1.77-1.36 (m, 16H), 1.28 (s, 3H), 1.26-1.07 (m, 2H), 0.60 (s, 3H) ppm; LCMS mlz 345 [M+H-H2Ot. Step 10. Preparation of compound Cll. Hydrogen bromide (5 drops, 48% in water) was added to a solution of (ClO, 2.15 g, 5.9 rnmol) in anhydrous methanol (150 mL) at room temperature in the dark under nitrogen, after which bromine (0.6 mL, 12 mmol) was added and the mixture was stirred for 90 min. The mixture was poured into ice-water (250 mL) and treated with 2N sodium hydroxide solution (20 mL) followed by saturated aqueous sodium bicarbonate solution (100 mL). The solids were collected under reduced pressure and purified by column chromatography on silica gel, eluting with hexanes / ethyl acetate (1:1), to provide compound Cll as a white solid (1.4 g, 53%): LCMS mlz 442 [M+Ht. Example 17. Alternative Preparation oflntermediate C9. 0 HO) r;½ Pd / C, H2 (50 psi) 0 ~---..,, • THF, 45 °C, overnight 0 A1 C21 Step 1 EtPPh3Br, t-BuOK THF,60°C Step4 H H C2 C22 Me2SO4,KOH 25°C, 16h Step 2 C20 2). 10% NaOH, 30 %H2O2, 25 °C Step5 MeMgBr, MAD THF,-78 °C Step3 Step I. Preparation of Compound C2. To a solution of PdiC ( 1 g, 10 % wet) in THF ( 10 mL) was added a solution of Al (10 g, 33.07 mmol) in dry THF (140 mL) was added in the mixture. After TLC showed the starting material was consumed completely, the mixture was filtered with CH2Ch (300 mL) and concentrated. The residue was purified by column chromatograph on silica 94 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 gel (PE:EA=8:l-4:1-2:1-l:1-EA) to give C2 (8.3 g, 82.43 %) as a white solid. 1H NMR (400 MHz, CDC13) o (ppm)= 3.96 (d, J=8.0 Hz, lH), 3.69 (d, J=8.0 Hz, IH), 2.69-2.65 (m, lH), 2.45- 2.29 (m, 4H), 2.12-1.69 (m, 8H), 1.63-1.23 (m, 7H), 0.88 (s, 3H) Step 2. Preparation of Compound C20. To a solution of compound C2 ( 15 g, 49.3 mmol) in THF (150 mL) was added KOH (8.4 g, 149.7 mmol) and Me2SO4 (12.9 g, 100.67 mol) at O °C. Then the mixture was wanned to 25 °C and stirred at the same temperature for 3 h. TLC (PE:EA=l:4) showed that the starting material was almost consumed. The mixture was quenched with the addition of 300 mL of water. The resulting solution was extracted with EtOAc (200 mL *3). The combined organic layers was washed with saturate aqueous NaCl (50 mL), dried over anhydrous Na2SO4 and evaporated in vacuum to give crude product, which was purified by column chromatography on silica gel (petroleum ether / ethyl acetate =4 / 1) to afford compound C20 (9.5 g, 60.5%) as a white solid. Step 3. Preparation of Compound C21. To a solution of compound 2,6-di-tert-butyl-4- methylphenol (4.15 g, 18.84 mmol) in toluene (8 mL) was added AlMe3 (4.7 mL, 9.42 mmol, 2 Min toluene) dropwise below 25 °C.The solution was stirred at room temperature for l h. Then a solution of compound C20 (1 g, 3 .14 mmol) in toluene (3 mL) was added dropwise at -78°C. After stirring at the same temperature for 1 h, MeMgBr (5.23 mL, 15.7 mmol, 3M in ethyl ether) was added dropwise at -78°C. The resulting solution was stirred at -78°C to -50°C for 3 h. TLC (PE / EtOAc = 1 / 1) showed the reaction was complete. The reaction was quenched by saturated aqueous NH4Cl (200 mL) at -78°C. The resulting mixture was filtered through a celite pad and the pad was washed with EtOAc (100 mL). The combined organic layer was separated, washed with brine (100 mL x 2) and concentrated in vacuum. The crude product was purified by a silica column chromatography (petroleum ether / ethyl acetate =4 / 1) to afford compound C21 (1 g, 95%) as a pale yellow oil. Step 4. Preparation of Compound C22. To a solution of PPh3EtBr (42.17 g, 113.6 mmol) in THF (40 mL) was added a solution oft-BuOK (12.75 g, 113.6 mmol) in THF (40 mL) at O 0C. After stirring at 60 °C for I h, a solution of compound C21 (7.6 g, 22. 72 11111101) in THF ( 40 mL) was added dropwise at 60 °C. Then the reaction mixture was stirred at 60 °C for 8 h. TLC (PE / EtOAc = 3 / 1) showed the starting material was also remained. To a solution of PPh3EtBr 95 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 (42.17 g, 113.6 mmol) in THF (40 mL) was added a solution oft-BuOK (12.75 g, 113.6 mmol) in THF (40 mL) at O °C. After stirring at 60 °C for I h. The solution was added to the reaction mixture. Then the reaction mixture was stirred at 60 °C for 8 h. TLC (PE / EA=3 / l) showed the starting material was also remained and the reaction was nearly unchanged. The reaction mixture was filtered and the filtrate was concentrated in vacuum to remove most of the solvent. The residue was partitioned between EtOAc (300 mL) and water (100 mLx3). The organic layer was washed with brine ( 100 mL), dried over Na2SO4 and concentrated in vacmm1. The cmde product was purified by silica column (PE:EA=5:l) to give compound C22 (4.0 g, 50.8%) as a pale yellow oil. 1H NMR (400 MHz, CDCb) 8 5.15-5.09(m, lH), 3.58(d, J=9.2Hz, lH), 3.49(s, lH), 3.33(s, 3H),3.20(d, .J=8.8Hz, lH), 2.40-1. lO(m, 28H), 0.85(s, 3H). Step 5. Preparation of Compound C9. To a solution of compound C22 (2.5 g, 7.21 rnmol) in THF (30 mL) was added dropwise a solution ofBH3-Me2S (7.21 rnL, 32.88 mmol) at O 0C. The solution was stirred at 25 °C for 3 h. TLC (PE / EtOAc = 1 / 1) showed the reaction was complete. After cooling to O °C, a solution ofNaOH (27.5 rnL, 3M) was added very slowly. After the addition was complete, H2O2 (15 mL, 30%) was added slowly and the inner temperature was maintained below 10 °C. The resulting solution was stirred at room temperahtre for 2 h. The resulting solution was extracted with EtOAc (100 mL x3). The combined organic solution was washed with sah1rated aqueous Na2S2O3 (100 mL), brine (100 mL), dried over Na2SO4 and concentrated in vacuum to give the crude product compound C23 (2.5 g, 95.15%) as a white solid. The cmde product was used for the next step without further purification. Example 18. General Procedure E: Preparation of A / B-cis scaffold C-21 analogs 96 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 0 NY~ -J(l-~ ""-I Ho' F H 73 0 Meo NS)F Br 5-fluorobenzotriazole t-L \ p 'N K2C03, THF, rt HO H 74 H 0 t\l_N- \ u,,f F N Ho' H 75 Preparation of compounds 73, 74, and 75. 5-Fluoro-lH-benzo[d][ l ,2,3]triazole (112 mg, 0.82 mmol) and potassium carbonate (3 73 mg, 2. 7 mmol) were added to a solution of compound Cll (120 mg, 0.27 mmol) in anhydrous tetrahydroforan (12 mL) at room temperature under nitrogen and the mixture was stirred for 16 h, at which point TLC analysis of the mixture (2:1 hexanes / ethyl acetate) indicated completion of the reaction. The mixture was diluted with water (80 mL) and extracted with ethyl acetate (3 x 80 mL). The combined organic extracts were washed with saturated aqueous sodium chloride solution (2 x 25 mL), dried with sodium sulfate and filtered. The solvents were removed under reduced pressure and the residue was semipurified by column chromatography on silica gel, eluting with hexanes / ethyl acetate (3:1), to provide a mixture of the three regioisomers. The residue was forther purified by reverse phase preparative HPLC to provide 73 as a white solid (44 mg, 33%): mp 82-84 °C; lH NMR (500 MHz, CDCb) 8 7.86 (dd, J = 9.0, 4.5 Hz, IH), 7.46 (dd, J = 9.0, 2.5 Hz, lH), 7.20 (ddd, J = 9.0, 9.0, 2.0 Hz, lH), 5.50 (d, JAB= 17.5 Hz, IH), 5.46 (d, JAB= 17.0 Hz, IH), 3.54 (d, J= 9.5 Hz, lH), 3.34 (s, 3H), 3.21 (d, J= 9.0 Hz, lH), 2.64 (t, J= 9.0 Hz, lH), 2.27-2.18 (m, IH), 2.18- 2.11 (m, lH), 1.96-1.88 (m, 2H), 1.83-1.40 (m, 12H), 1.39-1.10 (m, I0H), 0.73 (s, 3H) ppm; ESI MS mlz 496 [M-Hr. 97 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Further elution provided 75 as a white solid (25 mg, 18%): mp 205-207 °C; 1H NMR (500 MHz, CDCh) o 7.72-7.68 (m, lH), 7.31-7.27 (m, 2H), 5.43 (d, .h\B = 18.0 Hz, IH), 5.36 (d, JAB= 18.0 Hz, IH), 3.54 (d, J = 9.0 Hz, lH), 3.34 (s, 3H), 3.23 (d, J = 9.0 Hz, lH), 2.70 (t, J = 9.0 Hz, lH), 2.27-2.18 (m, lH), 2.18-2.12 (m, IH), 1.97-1.88 (m, 2H), 1.84-1.72 (m, 3H), 1.70-1.58 (m, 3H), 1.57-1.43 (m, 6H), 1.40-1.12 (m, IOH), 0.71 (s, 3H) ppm; ESI MS m / z 498 [M+Ht, Fmther elution provided 74 as an off-white solid (30 mg, 22%): mp 195-197 °C; 1H NMR (500 MHz, CDCh) 8 8.04 (dd, J= 9.0, 4.5 Hz, lH), 7.15 (dt, J= 9.0, 2.5 Hz, lH), 6.97 (dd, J= 7.5, 2.0 Hz, lH), 5.40 (d, JAB= 18.0 Hz, lH), 5.33 (d, JAB= 18.0 Hz, lH), 3.54 (d, J= 9.0 Hz, lH), 3.34 (s, 3H), 3.22 (d, J= 9.0 Hz, lH), 2.70 (t, J= 9.0 Hz, lH), 2.27-2.18 (m, lH), 2.18-2.12 (m, lH), 1.97-1.89 (m, 2H), 1.84-1.72 (m, 3H), 1.71-1.57 (m, 3H), 1.57-1.42 (m, 6H), 1.40-1.12 (m, l0H), 0.72 (s, 3H) ppm; ESI MS m / z 498 [M+Ht. Example 19. Preparation of compound 27. Br 4-CN-1-pyrazole K2C03, THF, rt Prepared according General Procedure E, Step 2 from compom1d Cll (60 mg, 0.14 mmol) and lH-pyrazole-4-carbonitrile (63 mg, 0.67 mmol), with purification by column chromatography on silica gel to provide compound 27 as an off-white solid (27.3 mg, 44%): mp 176-178 °C; 1HNMR (300 MHz, CDCb) o 7 .83 ( d, J = 12.3 Hz, 2H), 4.95 ( q, J = 18.3 Hz, 2H), 3 .53 ( d, J = 9.0 Hz, lH), 3.33 (s, 3H), 3.22 (d, J= 9.0 Hz, lH), 2.59 (t, J= 9.3 Hz, lH), 2.26-1.35 (m, 17H), 1.31-1.08 (m, 9H), 0.66 (s, 3H) ppm; ESI MS m / z 437 [M+H-H2Ot, Example 20. Preparation of compound 28. Br 4-Me-1-pyrazole H 98 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Prepared according General Procedure E, Step 2 from compound Cll (80 mg, 0.18 mmol) and 4-methyl-lH-pyrazole (45 mg, 0.54 mmol) with the substitution of cesium carbonate (177 mg, 0.54 mmol) in anhydrous acetonitrile (6 mL) at 65 °C, with purification by column chromatography on silica gel to provide compound 28 as a white solid (58 mg, 73%): mp 158- 160 °C; 1H NMR (500 MHz, CDCb) o 7.34 (s, lH), 7.16 (s, lH), 4.86 (d, IAB = 17.5 Hz, IH), 4.78 (d, JAB= 18.0 Hz, lH), 3.54 (d, J = 9.0 Hz, IH), 3.32 (s, 3H), 3.19 (d, J = 9.0 Hz, IH), 2.54 (t, J = 9.5 Hz, lH), 2.23-2.13 (m, lH), 2.09 (s, 3H), 2.07-2.02 (m, lH), 1.97-1.87 (m, 2H), 1.80-1.35 (m, 12H), 1.33-1.10 (m, lOH), 0.66 (s, 3H) ppm; ESI MS mlz 443 [M+Hf. Example 21. Preparation of compounds 29 and 30. Br 5-Me-tetrazole Prepared according General Procedure E, Step 2 from compound Cll (100 mg, 0.23 mmol) and 5-methyltetrazole (95 mg, 1.13 mmol), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 29 as an off-white solid (12.2 mg, 12%): mp 90-92 °C; 1HNMR (300 MHz, CDCb) 8 5.09 (q,J= 18.0 Hz, 2H), 3.53 (d, J= 9.0 Hz, lH), 3.33 (s, 3H), 3.22 (d, J = 9.0 Hz, lH), 2.65 (t, J = 9.0 Hz, lH), 2.47 (s, 3H), 2.25-1.58 (m, 9H), 1.55-1.14 (m, 17H), 0.67 (s, 3H) ppm; ESI MS mlz 428 [M+H-H2Of. Fm1her elution provided 30 as an off-white solid (13.4 mg, 13%): mp 70-72 °C; 1HNMR (300 MHz, CDCb) o 5.34 (s, 2H), 3.54 (d, J = 9.0 Hz, lH), 3.33 (s, 3H), 3.20 (d, J = 9.0 Hz, IH), 2.64-2.57 (m, 4H), 2.43-1.91 (m, 6H), 1.81-1.10 (m, 20H), 0.70 (s, 3H) ppm; ESI MS mlz 428 [M+H-H2Of. Example 22. Preparation of compounds 31 and 32. 99 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 0 Br N---N N---N ~ \ triazole / J N N" + K2C03, THF, rt 31 32 H Hd H H Prepared according General Procedure E, Step 2 from compow1d Cll (82 mg, 0.18 mmol) and lH-1,2,3-triazole (75 mg, 1.08 mmol), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 32 as an off-white solid ( 17 mg, 22%): mp 80-83 °C; 1H NMR (500 MHz, CDCh) o 7.79 (s, lH), 7.66 (s, lH), 5.26 (d, J,AJ3 = 18.0 Hz, 1H), 5.13 (d, JAB= 18.0 Hz, lH), 3.53 (d, J = 9.0 Hz, IH), 3.33 (s, 3H), 3.20 (d, J = 9.0 Hz, lH), 2.64 (t, J = 9.0 Hz, lH), 2.26-2.16 (m, lH), 2.12-2.06 (m, lH), 1.96-1.70 (m, 6H), 1.66-1.42 (m, 6H), 1.35-1.10 (m, llH), 0.90-0.83 (m, lH), 0.66 (s, 3H) ppm; ESI MS mlz 430 [M+Ht. Further elution provided 31 as an off-white solid (12 mg, 16%): mp 71-74 °C; 1H NMR (500 MHz, CDCh) o 7.68 (s, 2H), 5.24 (d, J_AI3 = 17.5 Hz, lH), 5.21 (d,JAB = 17.5 Hz, lH), 3.54 (d, J = 9.0 Hz, lH), 3.33 (s, 3H), 3.19 (d, J = 9.0 Hz, lH), 2.56 (t, J = 9.0 Hz, IH), 2.24-2.15 (m, 1H), 2.11-2.05 (m, lH), 1.96-1.88 (m, 2H), 1.82-1.68 (m, 3H), 1.66-1.40 (m, 6H), 1.40-1.09 (m, l lH), 0.90-0.83 (m, lH), 0.70 (s, 3H) ppm; ESI MS mlz 430 [M+Ht, Example 23. Preparation of compound 33. Br 4-S02Me-1-pyrazole K2C03, THF, rt Prepared according General Procedure E, Step 2 from compow1d Cll (80 mg, 0.18 mmol) and 4-(methylsulfonyl)-lH-pyrazole (79 mg, 0.54 rnmol), with semi-purification by coltmm chromatography on silica gel followed by reverse phase preparative HPLC to provide 33 as a white solid (62 mg, 69%): mp 110-112 °C; 1H NMR (500 MHz, CDCb) o 7.92 (s, lH), 7.86 (s, IH), 5.00 (d, .l.!\B = 18.0 Hz, lH), 4.90 (d, JAB= 17.5 Hz, lH), 3.53 (d, J= 9.0 Hz, lH), 3.33 (s, 3H), 3.21 (d, J = 9.0 Hz, lH), 3.13 (s, 3H), 2.60 (t, J = 9.0 Hz, lH), 2.25-2.15 (m, lH), 2.08- 100 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 2.02 (m, IH), 1.96-1.88 (m, 2H), 1.82-1. 71 (m, 3H), 1.67-1.56 (m, 3H), 1.54-1.41 (m, 6H), 1.39-1.13 (m, 10H), 0.67 (s, 3H) ppm; ESI MS ml= 505 [M-H]". Example 24. Preparation of compound 34. Br 4-Cl-1-pyrazole K2C03, THF, rt H Prepared according General Procedure E, Step 2 from compound Cll (80 mg, 0.18 mmol) and 4-chloro-IH-pyrazole (45 mg, 0.54 mmol), with purification by column chromatography on silica gel to provide compound 34 as a white solid (58 mg, 70%): mp 163-165 °C; 1H NMR (500 MHz, CDCb) 8 7.45 (s, IH), 7.41 (s, lH), 4.99 (d, JAB= 17.5 Hz, lH), 4.80 (d, JAB= 17.5 Hz, lH), 3.53 (d, J = 9.0 Hz, lH), 3.33 (s, 3H), 3.21 (d, J = 9.0 Hz, lH), 2.56 (t, J = 9.0 Hz, lH), 2.24-2.14 (m, lH), 2.07-2.01 (m, 1H), 1.96-1.88 (m, 2H), 1.80-1.69 (m, 3H), 1.66-1.35 (m, 9H), 1.34-1.10 (m, lOH), 0.66 (s, 3H) ppm; ESI MS mlz 463 [M+Ht. Example 25. Preparation of compound 38. 0 A11 Br pyrazolo[4,3-b]pyridine K2C03, THF, rt Prepared according General Procedure B from compound All (31 mg, 0.071 mmol) and 2Hpyrazolo[ 4,3-b ]pyridine (168 mg, 1.41 mmol), with purification by reverse phase preparative HPLC to provide 38 as a white solid (8. 7 mg, 25%): mp 153-154 °C; 1HNMR (500 MHz, CDCb) 8 8.60 (d, J = 3.5 Hz, lH), 8.28 (s, IH), 7.59 (d, J = 8.5 Hz, lH), 7.29 (dd, J = 8.5, 4.5 Hz, IH), 5.16 (dd, J= 29.0, 18.0 Hz, 2H), 3.48 (d, J= 10.0 Hz, lH), 3.38 (d, J= 10.0 Hz, lH), 3.30 (s, 3H), 2.66 (t, J= 9.0 Hz, IH), 2.22-2.00 (m, lH), 2.13-2.11 (m, IH), 2.10-2.08 (m, lH), 1.79-1.40 (m, l lH), 1.33-1.11 (m, lOH), 0.99-0.97 (m, IH), 0.88-0.86 (m, IH), 0.73 (s, 3H) ppm; ESI MS m / z 480 [M+Ht. 101 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 Example 26. Preparation of compounds 39 and 40. Meo, ~Br H~ benzotriazole + PCT / US2014 / 052417 0 N----N ~~\\ -. - HO H 40 Prepared according General Procedure B from compound All (21 mg, 0.047 mmol) and lHbenzotriazole (33 mg, 0.28 mmol), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 39 as an off-white solid ( 13 mg, 58%): mp 78-80 °C; 1H NMR (500 MHz, CDCh) 8 8.11-8.07 (m, lH), 7.51-7.46 (m, IH), 7.40-7.36 (m, lH), 7.33 (d, J= 7.5, lH), 5.41 (s, 2H), 3.48 (d, J= 10.0 Hz, lH), 3.38 (d, J= 10.0 Hz, lH), 3.30 (s, 3H), 2.71 (t, J= 9.0 Hz, lH), 2.27-2.18 (m, lH), 2.17-2.10 (m, lH), 2.08-2.02 (m, lH), 1.82-1.68 (m, 4H), 1.67-1.42 (m, 7H), 1.35-1.09 (m, l0H), 1.05-0.95 (m, lH), 0.91- 0.83 (m, lH), 0.76 (s, 3H) ppm; ESI MS mlz 480 [M+Ht. Further elution provided 40 as an off-white solid (7 mg, 30%): mp 70-72 °C; 1H NMR (500 MHz, CDCb) o 7.90-7.85 (m, 2H), 7.41-7.36 (m, 2H), 5.54 (d, JAB= 17.0 Hz, lH), 5.48 (d, JAB = 17.0 Hz, lH), 3.48 (d, J= 10.0 Hz, lH), 3.38 (d, J= 10.0 Hz, lH), 3.290 (s, 3H), 2.63 (t, J= 9.0 Hz, lH), 2.28-2.20 (m, lH), 2.17-2.12 (m, lH), 2.07-2.01 (m, lH), 1.80-1.68 (m, 4H), 1.67-1.46 (m, 6H), 1.45-1.36 (m, lH), 1.35-1.08 (m, lOH), 1.04-0.94 (m, lH), 0.90-0.82 (m, lH), 0.77 (s, 3H) ppm; ESI MS m / z 480 [M+Ht. Example 27. Preparation of compounds 41, 42, and 43. MoO, ~~ _5-f_lu_oro_b_en_zo_tri_az_ol_e ~-✓ K2CO3, THF, rt HO H A11 Prepared according General Procedure B from compound All (100 mg, 0.23 mmol) and 5- fluorobenzotriazole (124 mg, 0.91 mmol), with purification by reverse phase preparative HPLC 102 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 to provide 43 as an off-white solid (39.2 mg, 40%): mp 55-60 °C; 1HNMR (300 MHz, CDCb) 8 7.86 (dd, J= 4.8, 4.5 Hz, lH), 7.46 (dd, J= 9.0, 2.1 Hz, lH), 7.23-7.16 (m, lH), 5.49 (q, J= 18.0 Hz, 2H), 3.48 (d, J = 9.9 Hz, lH), 3.38 (d, J= 9.9 Hz, lH), 3.37 (s, 3H), 2.66 (t, J= 8.7 Hz, lH), 2.29-2.01 (m, 3H), 1.79-1.50 (m, 15H), 1.45-1.05 (m, 6H), 1.01-0.82 (m, 2H), 0.76 (s, 3H) ppm; ESI MS mlz 498 [M+Ht. Fmther elution provided 41 as an off-white solid (21.2 mg, 34%): mp 65-70 °C; 1HNMR (300 MHz, CDCb) 8 8.03 (dd, J = 9.0, 4.5 Hz, IH), 7.18-7.12 (m, lH), 6.99-6.58 (m, IH), 5.37 (q, J = 18.3 Hz, 2H), 3.47 (d, J= 10.2 Hz, lH), 3.38 (d, J= 10.2 Hz, lH), 3.31 (s, 3H), 2.72 (t, J= 9.0 Hz, lH), 2.28-2.02 (m, 3H), 1.79-1.33 (m, 13H), 1.29-0.84 (m, IOH), 0.75 (s, 3H) ppm; ESI MS mlz 498 [M+Ht. Further elution provided 42 as an off-white solid (21.2 mg, 34%): mp 60-65 °C; 1HNMR (300 MHz, CDCh) 8 7.73 (d, J= 9.9 Hz, lH), 7.35-7.21 (m, 2H), 5.40 (q, J= 18.3 Hz, 2H), 3.49 (d, J = 9.9 Hz, lH), 3.37 (d, J = 10.2 Hz, lH), 3.30 (s, 3H), 2.72 (t, J = 9.0 Hz, lH), 2.28-2.03 (m, 3H), 1.82-1.46 (m, 9H), 1.37-1.16 (m, 12H), 1.06-0.84 (m, 2H), 0.74 (s, 3H) ppm; ESI MS mlz 498 [M+Ht. Example 28. Preparation of compounds 44 and 45. 1 H-pyrazolo[ 3,4-c]pyridine K2C03, THF, rt Prepared according General Procedure B from compound All (50 mg, 0.11 mmol) and 1Hpyrazolo[ 3,4-c]pyridine (67 mg, 0.57 mmol), with purification by reverse phase preparative HPLC to provide 44 as an off-white solid (7.5 mg, 14%): mp 160-162 °C; 1HNMR (300 MHz, CDCl3) 8 9 .26 (s, lH), 8.17 ( d, J = 6.0 Hz, lH), 7 .97 ( d, J = 0.9 Hz, lH), 7.53 (dd, J = 6.0, 1.2 Hz, lH), 5.27 (q, J = 18.0 Hz, 2H), 3.46 (d, J = 9.9 Hz, 2H), 3.37 (d, J = 10.2 Hz, 2H), 2.68 (t, J = 9.3 Hz, lH), 2.30-2.19 (m, lH), 1.18-2.01 (m, 3H), 1.79-1.64 (m, 5H), 1.61-1.45 (m, 7H), 1.39-1.08 (m, SH), 1.05-0.84 (m, 3H), 0.74 (s, 3H) ppm; ESI MS rnlz 480 [M+Ht. 103 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Further elution provided 45 as an off-white solid (14.2 mg, 26%): mp 92-94 °C; 1H NMR (300 MHz, CDCh) 8 8.80 (s, lH), 8.34 (d, J= 5.4 Hz, lH), 8.09 (d, J= 0.6 Hz, lH), 7.64 (dd, J= 5.7, 1.2 Hz, IH), 5.25 (q, J= 18.0 Hz, 2H), 3.48 (d, J= 9.9 Hz, 2H), 3.38 (d, J = 9.9 Hz, 2H), 3.31 (s, 3H), 2.70 (t, J= 8.7 Hz, lH), 2.27-1.98 (m, 3H), 1.81-1.44 (m, lOH), 1.34-1.12 (m, 9H), 1.10- 0.81 (m, 2H), 0.74 (s, 3H) ppm; ESI MS mlz 480 [M+Ht. Example 29. Preparation of compound 46. Br benzimidazole Prepared according General Procedure B from compom1d All (50 mg, 0.114 mmol) and benzimidazole (268 mg, 2.3 mmol), with purification by reverse phase preparative HPLC to provide 46 as a white solid (36.5 mg, 67%): mp 104-105 °C; 1HNMR (500 MHz, CDCh) 8 7.99 (s, lH), 7.86-7.82 (m, lH), 7.33-7.29 (m, 2H), 7.20-7.18 (m, lH), 4.93 (dd, J= 24.0, 18.5 Hz, 2H), 3.49 (d, J = 10.0 Hz, IH), 3.38 (d, J = 10.0 Hz, lH), 3.30 (s, 3H), 2.66 (t, J = 9.0 Hz, lH), 2.24-2.22 (rn, lH), 2.07-2.05 (m, lH), 1.80-1.42 (m, llH), 1.34-1.11 (m, llH), 0.99-0.98 (m, IH), 0.88-0.86 (rn, lH), 0.73 (s, 3H) ppm; ESI MS mlz 479 [M+Hr. Example 30. Preparation of compounds 47, 48, and 49. -~ ~-~ _4-'_f_luo_ro_be_nz_ot_ria_zo_le_ ~ K2C03, THF, rt HO H A11 Prepared according General Procedure B from compound All (100 mg, 0.227 mmol) and 4- fluorobenzotriazole (311 mg, 2.27 mmol), with purification by reverse phase preparative HPLC to provide 49 as a white solid (22.5 mg, 20%): mp 125-126 °C; 1HNMR (500 MHz, CDCb) 8 104 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 7.67 (d, J = 8.5 Hz, lH), 7.32-7.30 (m, IH), 7.03 (dd, J = 10.5, 7.5 Hz, lH), 5.54 (dd, J = 28.0, 17.0 Hz, 2H), 3.48 (d, J = 10.0 Hz, IH), 3.38 (d, J = 10.0 Hz, lH), 3.30 (s, 3H), 2.67 (t, J = 9.0 Hz, lH), 2.24-2.22 (m, IH), 2.15-2.13 (m, lH), 2.05-2.03 (m, lH), 1.78-1.50 (m, 9H), 1.42-1.40 (m, lH), 1.34-1.12 (m, 11H), 0.99-0.98 (m, IH), 0.87-0.86 (m, IH), 0.77 (s, 3H) ppm; ESI MS mlz 498 [M+Hf. Fmther elution provided 47 as a white solid (7.3 mg, 6%): mp 83-84 °C; 1Hm1R (500 MHz, CDCb) 8 7.86 (d, J = 8.0 Hz, lH), 7.29-7.27 (m, lH), 7.13 (dd, J = 10.5, 7.5 Hz, lH), 5.54 (s, 2H), 3.49 (d, J= 10.0 Hz, IH), 3.38 (d, J= 10.0 Hz, lH), 3.30 (s, 3H), 2.70 (t, J= 9.0 Hz, lH), 2.24-2.22 (m, lH), 2.15-2.13 (m, lH), 2.06-2.04 (m, lH), 1.79-1.73 (111, 4H), 1.63-1.43 (m, 7H), 1.34-1.13 (m, l0H), 1.02-1.00 (m, lH), 0.89-0.87 (m, lH), 0.75 (s, 3H) ppm; ESI MS mlz 498 [M+Ht. Further elution provided 48 as a white solid (26.3 mg, 23%): mp 129-130 °C; 1HNMR (500 MHz, CDCh) 8 7.42 (td, J= 8.0, 4.5 Hz, lH), 7.10 (d, J= 8.5 Hz, lH), 7.04 (dd, J= 10.0, 7.5 Hz, lH), 5.42 (s, 2H), 3.49 (d, J = 10.0 Hz, lH), 3.38 (d, J = 10.0 Hz, lH), 3.30 (s, 3H), 2.70 (t, J = 9.0 Hz, lH), 2.23-2.21 (m, lH), 2.15-2.13 (m, lH), 2.06-2.04 (m, IH), l.78-1.46 (m, l0H), 1.34-1.12 (m, l IH), 1.01-0.99 (m, lH), 0.89-0.88 (m, lH), 0.74 (s, 3H) ppm; ESI MS mlz 498 [M+H]+. Example 31. Preparation of compounds 50 and 51. 5-methoxybenzotriazole Prepared according General Procedure B from compound All (140 mg, 0.32 mmol) and 5- methoxybenzotriazole (132 mg, 0.89 mmol), with purification by reverse phase preparative HPLC to provide 50 as a white solid (12.9 mg, 8%): mp 165-166 °C; 1HNMR (500 MHz, CDCb) 8 7.72 (d, J = 10.0 Hz, lH), 7.07-7.05 (m, 2H), 5.43 (dd, J = 29.5, 17.0 Hz, 2H), 3.87 (s, 3H), 3.47 (d, J = 10.0 Hz, lH), 3.38 (d, J = 10.0 Hz, lH), 3.29 (s, 3H), 2.63 (t, J = 9.0 Hz, IH), 2.24- 105 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 2.22 (m, lH), 2.14-2.12 (m, lH), 2.05-2.04 (m, lH), 1.77-1.69 (m, 4H), 1.62-1.10 (m, 17H), 0.99-0.98 (m, lH), 0.86-0.84 (m, lH), 0.76 (s, 3H) ppm; ESI MS mlz 510 [M+Ht. Further elution provided 51 as a white solid ( 11.8 mg, 7%): mp 106-107 °C; 1HNMR (500 MHz, CDCh) & 7.93 (d, J= 9.0 Hz, lH), 7.02 (dd, J= 9.0, 2.0 Hz, IH), 6.61 (d, J= 2.0 Hz, lH), 5.33 (d, J = 3.5 Hz, 2H), 3.88-3.86 (m, 3H), 3.48 (d, J = 10.0 Hz, lH), 3.38 (d, J = 10.0 Hz, lH), 3.30 (s, 3H), 2.70 (t, J = 9.0 Hz, lH), 2.23-2.21 (m, lH), 2.15-2.13 (m, lH), 2.06-2.04 (m, lH), 1.76- 1.70 (m, 4H), 1.64-1.44 (m, 6H), 1.34-1.13 (m, 1 IH), 0.99-0.98 (m, lH), 0.89-0.87 (m, IH), 0.75 (s, 3H) ppm; ESI MS mlz 510 [M+Ht. Example 32. Preparation of compounds 52, 53, and 54. 4,5-<:lifluorobenzotriazole K2C03, THF, rt Prepared according General Procedure B from compound All (100 mg, 0.23 mmol) and 4,5- difluorobenzotriazole (352 mg, 2.3 mmol), with purification by reverse phase preparative HPLC to provide 54 as a white solid (46.0 mg, 32%): mp 86-87 °C; 1HNMR (500 MHz, CDCb) b 7.63 (ddd, J = 9.0, 3.5, 1.0 Hz, IH), 7.30-7.28 (m, lH), 5.53 (dd, J = 31.5, 17.0 Hz, 2H), 3.48 (d, J = 10.0 Hz, IH), 3.38 (d, J = 10.0 Hz, lH), 3.30 (s, 3H), 2.65 (t, J = 9.0 Hz, lH), 2.24-2.22 (m, IH), 2.14-2.12 (m, lH), 2.06-2.04 (m, lH), 1.78-1.73 (m, 4H), 1.63-1.41 (m, 7H), 1.34-1.12 (m, lOH), 1.03-1.00 (m, lH), 0.88-0.86 (m, lH), 0.77 (s, 3H) ppm; ESI MS mlz 498 [M+H-H2Ot. Further elution provided 52 as a white solid (19.3 mg, 13%): mp 82-83 °C; 1HNMR (500 MHz, CDCh) 8 7.80 (ddd, J = 9.0, 3.5, 1.0 Hz, lH), 7.23-7.21 (m, lH), 5.52 (s, 2H), 3.49 (d, J = 10.0 Hz, lH), 3.38 (d, J = 10.0 Hz, lH), 3.30 (s, 3H), 2.70 (t, J = 9.0 Hz, lH), 2.23-2.21 (m, lH), 2.14-2.12 (m, lH), 2.07-2.05 (m, lH), 1.79-1.72 (m, 4H), 1.64-1.44 (m, 7H), 1.35-1.13 (m, I0H), 1.02-1.01 (m, lH), 0.89-0.88 (m, lH), 0.75 (s, 3H) ppm; ESI MS rnlz 516 [M+Ht. 106 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Further elution provided 53 as a white solid (34.3 mg, 24%): mp 144-145 °C; 1HNMR (500 MHz, CDCh) 8 7.38-7.37 (m, IH), 7.04 (ddd, J = 9.0, 3.0, 1.0 Hz, lH), 5.41 (dd, J = 22.5, 18.0 Hz, 2H), 3.49 (d, J= 10.0 Hz, lH), 3.38 (d, J= 10.0 Hz, lH), 3.30 (s, 3H), 2.71 (t, J= 9.0 Hz, lH), 2.24-7.22 (m, lH), 2.12.14-2.12 (m, lH), 2.07-2.05 (111, lH), 1.78-1.71 (m, 4H), 1.64-1.47 (m, 7H), 1.34-1.13 (m, l0H), 1.02-1.01 (m, lH), 0.89-0.88 (m, lH), 0.74 (s, 3H) ppm; ESI MS mlz 516 [M+H]+. Example 33. Preparation of compounds 55 and 56. Br 4,6-difluorobenzotriazole r~~N~"~ :)I. F :-- Ho' Fi 55 F Prepared according General Procedure B from compound All (125 mg, 0.28 mmol) and 4,6- difluoro-1H-benzo[d][l,2,3]triazole (219 mg, 1.60 11111101), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 55 as an off-white solid (28 mg, 19%): mp 182-186 °C; 1H NIVIR (300 MHz, CDCh) 8 7.30 (dd, J = 8.l, 1.5 Hz, lH), 6.90 (ddd, J= 9.9, 9.9, 2.1 Hz, lH), 5.55 (d, / o,.B = 17.1 Hz, lH), 5.47 (d, JAB= 17.1 Hz, lH), 3.48 (d, J = 9.9 Hz, lH), 3.37 (d, J = 10.2 Hz, lH), 3.30 (s, 3H), 2.67 (t, J = 8.7 Hz, IH), 2.30-2.17 (m, lH), 2.17-2.09 (m, lH), 2.09-1.99 (m, lH), 1.83-1.66 (m, 4H), 1.66-1.40 (m, 7H), 1.40-0.81 (m, 12H), 0.76 (s, 3H) ppm; ESI MS mlz 516 [M+Ht. Further elution provided 56 as an off-white solid (19 mg, 13%): mp 96-100 °C; 1H NMR (300 MHz, CDCb) 5 6.87 (ddd, J= 9.6, 9.6, 1.8 Hz, lH), 6.79 (ddd, J= 7.5, 2.1, 0.6 Hz, lH), 5.41 (d, JAB= 18.0 Hz, lH), 5.34 (d, .f.">.B = 18.3 Hz, lH), 3.49 (d, J = 9.9 Hz, lH), 3.38 (d, .J = 9.9 Hz, lH), 3.31 (s, 3H), 2.72 (t, J= 8.7 Hz, lH), 2.30-2.00 (m, 3H), 1.85-1.41 (m, llH), 1.38-0.82 (m, 12H), 0.74 (s, 3H) ppm; SI MS mlz 516 [M+Ht. Example 34. Preparation of compounds 57 and 58. 107 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 0 ' Br MeO 4-methoxybenzotriazole ..;: HO A A11 K2C03, THF, rt Prepared according General Procedure B from compound All (100 mg, 0.23 mmol) and 4- methoxybenzotriazole (675 mg, 4.5 mmol), with purification by reverse phase preparative HPLC to provide 58 as a light brown solid (27 mg, 23%): mp 78-80 °C; 1H NMR (500 MHz, CDCb) 8 7.39(t,J=8.0Hz, 1H),6.86(d,J=8.l Hz, 1H),6.70(d,.l=7.8Hz, 1H),5.37(dd,.J= 18.0,2.7 Hz, 2H), 4.12 (s, 3H), 3.48 (d, J = 10.0 Hz, lH), 3.38 (d, J = 10.1 Hz, lH), 3.30 (s, 3H), 2.68 (t, J = 9.0 Hz, lH), 2.24-2.18 (m, IH), 2.14-2.09 (m, lH), 2.06-2.02 (m, lH), 1.76-1.69 (m, 4H), 1.64-1.55 (m, 3H), 1.54-1.37 (m, 4H), 1.36-1.19 (m, l0H), 1.18-1.08 (m, 2H), 1.04-0.93 (m, lH), 0.90-0.84 (m, 2H), 0.75 (s, 3H) ppm; APCI MS mlz 510 [M+Hf. Further elution afforded 57 as an off-white solid (33 mg, 29%): mp 200-202 °C; 1H NMR (500 MHz, CDCl3) & 7.63 (d, J = 8.4 Hz, lH), 7.24 (d, J = 8.3 Hz, lH), 6.76 (d, J = 7.7 Hz, lH), 5.58 (dd, J = l 7.9, 9.3 Hz, 2H), 3.89 (s, 3H), 3.49 (d, J = 10.0 Hz, lH), 3.39 (d, .J = 10.0 Hz, lH), 3.30 (s, 3H), 2.67 (t, J= 8.9 Hz, lH), 2.25-2.18 (m, lH), 2.15-2.11 (m, IH), 2.06-2.02 (m, lH), 1.73- 1.67 (m, 3H), 1.65-1.47 (m, 7H), 1.42-L 11 (m, 12H), 1.04-0.95 (m, 1 H), 0.89-10.84 (m, 1 H), 0.75 (s, 3H) ppm; APCI MS mlz 510 [M+Ht. Example 35. Preparation of compounds 59, 60, and 61. 4-chlorobenzotriazole + 0 ~-0 N~ .. H N Cl - Hd. A 60 + 30 -~-u~, Cl N~ I ,..._ Hd H 61 Prepared according General Procedure B from compound All (150 mg, 0.34 mmol) and 4- chlorobenzotriazole (156 mg, 1.02 mmol), with semi-purification by column chromatography on 108 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 silica gel followed by reverse phase preparative HPLC to provide 61 as a light brown solid (64 mg, 37%): mp 170-172 °C; 1H NMR (500 MHz, CDCh) o 7.79 (dd, J= 8.5, 0.5 Hz, IH), 7.40 (dd, J= 7.0, 0.5 Hz, lH), 7.32 (dd, J= 8.5, 7.5 Hz, IH), 5.57 (d, JAB= 17.0 Hz, lH), 5.53 (d, JAB = 17.0 Hz, IH), 3.48 (d, J = 10.0 Hz, lH), 3.38 (d, J= 10.0 Hz, lH), 3.30 (s, 3H), 2.66 (t, J= 9.0 Hz, lH), 2.28-2.18 (m, lH), 2.17-2.10 (m, lH), 2.07-2.01 (m, lH), 1.81-1.68 (m, 4H), 1.66- 1.46 (m, 6H), 1.44-1.36 (m, lH), 1.35-1.08 (m, lOH), 1.04-0.94 (m, lH), 0.90-0.83 (m, lH), 0.77 (s, 3H) ppm; ESI MS mlz 514 [M+Hf. Further elution afforded 59 as a light brown solid (8 mg, 4%): mp 162-164 °C; 1H NMR (500 MHz, CDCb) & 7.99 (d, J = 8.5 Hz, lH), 7.43 (d, J = 7.5, lH), 7.29 (t, J = 7.5, lH), 5.71 (s, 2H), 3.48 (d, J = I0.0 Hz, lH), 3.38 (d, J = I0.0 Hz, lH), 3.30 (s, 3H), 2.72 (t, J = 9.0 Hz, IH), 2.27- 2.10 (m, 2H), 2.08-2.02 (m, lH), 1.85-1.37 (m, 11H), 1.35-1.09 (m, lOH), 1.05-0.95 (m, lH), 0.91-0.83 (m, IH), 0.76 (s, 3H) ppm; APCI MS mlz 514 [M+Hf. Further elution provided 60 as a light brown solid (32 mg, 18%): mp 105-107 °C; 1H NMR (500 MHz, CDCh) o 7.43-7.36 (m, 2H), 7.23 (dd, J = 7.5, 1.5 Hz, IH), 5.42 (s, 2H), 3.49 (d, J = 10.0 Hz, lH), 3.38 (d, J = 10.0 Hz, lH), 3.30 (s, 3H), 2.71 (t, J = 9.0 Hz, lH), 2.27-2.18 (m, lH), 2.18-2.10 (m, lH), 2.08-2.01 (m, lH), 1.82-1.68 (m, 4H), 1.67-1.42 (m, 7H), 1.35-1.09 (m, l0H), 1.05-0.95 (m, lH), 0.91-0.84 (m, lH), 0.75 (s, 3H) ppm; ESI MS mlz 514 [M+Hf. Example 36. Preparation of compounds 62, 63, and 64. 0 -, . H Ha° H A11 Br 4,5-dimethoxybenzotriazole 0 ,~- N;:,N + . 62 HO H + Prepared according General Procedure B from compound All (100 mg, 0.23 mmol) and 4,5- dimethoxy-1H-benzo[d][l,2,3]triazole (101 mg, 0.57 mmol), with semi-purification by column 109 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 chromatography on silica gel followed by reverse phase preparative HPLC to provide 64 as a light yellow solid (32 mg, 26%): mp 188-190 °C; 1H NMR (500 MHz, CDCb) 8 7.50 (d, J= 9.0 Hz, lH), 7.22 (d, J = 9.5 Hz, lH), 5.49 (d, J.@ = 17.0 Hz, IH), 5.43 (d, JAB= 17.0 Hz, lH), 4.27 (s, 3H), 3.95 (s, 3H), 3.48 (d, J = 10.0 Hz, IH), 3.38 (d, J = 10.0 Hz, IH), 3.29 (s, 3H), 2.63 (t, J = 8.5 Hz, lH), 2.27-2.17 (m, lH), 2.16-2.10 (m, lH), 2.07-2.01 (m, IH), 1.79-1.68 (m, 4H), 1.65-1.46 (m, 7H), 1.42-1.08 (m, IOH), 1.03-0.93 (m, lH), 0.89-0.81 (m, lH), 0.76 (s, 3H) ppm; APCI MS mlz 540 [M+Hf. Further elution provided 63 as a white solid (19 mg, 15%): mp 88-90 °C; 1H NMR (500 MHz, CDCh) 8 7.24 (d, J= 9.0 Hz, lH), 6.82 (d, J= 9.0 Hz, IH), 5.36 (d, JAB= 18.0 Hz, lH), 5.32 (d, JAB= 18.0 Hz, 1H), 4.57 (s, 3H), 3.93 (s, 3H), 3.49 (d, J = 10.0 Hz, IH), 3.38 (d, J = 10.0 Hz, lH), 3.30 (s, 3H), 2.69 (t, .J = 9.0 Hz, lH), 2.26-2.17 (m, IH), 2.16-2.10 (m, lH), 2.08-2.01 (m, lH), 1.80-1.68 (m, 4H), 1.66-1.41 (m, 7H), 1.36-1.09 (m, lOH), 1.04-0.94 (m, lH), 0.90-0.83 (m, lH), 0.74 (s, 3H) ppm; APCI MS mlz 540 [M+Ht. Further elution provided 62 as a white solid (13 mg, 11%): mp 110-112 °C; 1H NMR (500 MHz, CDCb) 8 7.75 (br s, lH), 7.11 (d, .J = 7.5 Hz, lH), 5.54 (d, .l.l\B = 18.0 Hz, lH), 5.50 (d, J.JIB = 18.0 Hz, IH), 3.96 (s, 3H), 3.90 (s, 3H), 3.48 (d, J = 10.0 Hz, IH), 3.38 (d, J = 10.0 Hz, lH), 3.30 (s, 3H), 2.69 (t, J = 8.5 Hz, lH), 2.28-2.01 (m, 3H), 1.83-1.68 (m, 4H), 1.65-1.47 (m, 6H), 1.44-1.37 (m, lH), 1.36-1.08 (m, lOH), 1.04-0.94 (m, lH), 0.90-0.83 (m, lH), 0.76 (s, 3H) ppm; APCI MS m / z 540 [M+Hf. Example 37. Preparation of compounds 65 and 66. 4,6-dimethoxybenzotriazole Prepared according General Procedure B from compound All (125 mg, 0.28 mmol) and 4,6- dimethoxy-1H-benzo[d][l,2,3]triazole (127 mg, 0.71 mmol), \vith purification by column chromatography on silica gel to provide 65 as a light yellow solid (42 mg, 28%): mp 106-108 110 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 °C; 1H NMR (500 MHz, CDCh) o 6.67 (d, J = 2.0 Hz, IH), 6.33 (d, J = 2.0 Hz, lH), 5.43 (d, JAB = 17.0 Hz, lH), 5.38 (d, J,IB = 17.0 Hz, lH), 3.98 (s, 3H), 3.96 (s, 3H), 3.47 (d, J = 10.0 Hz, lH), 3.38 (d, J= 10.0 Hz, IH), 3.29 (s, 3H), 2.61 (t, J= 9.0 Hz, lH), 2.26-2.16 (m, lH), 2.13-2.07 (m, lH), 2.07-2.00 (m, lH), 1.77-1.67 (m, 4H), 1.63-1.46 (m, 7H), 1.39-1.08 (m, 10H), 1.02- 0.93 (m, lH), 0.88-0.80 (m, lH), 0.75 (s, 3H) ppm; APCI MS mlz 540 [M+Ht. Fmther elution provided 66 as an off-white solid (52 mg, 34%): mp 110-112 °C; 1H NMR (500 MHz, CDCb) o 6.34 (d, J= 2.0 Hz, IH), 6.16 (d, J= 2.0 Hz, IH), 5.28 (s, 2H), 4.06 (s, 3H), 3.84 (s, 3H), 3.48 (d, J = I 0.0 Hz, lH), 3.38 (d, J = 10.0 Hz, lH), 3.29 (s, 3H), 2.66 (t, J = 9.0 Hz, lH), 2.25-2.16 (m, lH), 2.13-2.08 (m, lH), 2.07-2.01 (m, lH), 1.79-1.67 (m, 4H), 1.64-1.46 (m, 6H), 1.45-1.37 (m, lH), 1.35-1.08 (m, l0H), 1.03-0.93 (m, IH), 0.89-0.82 (m, lH), 0.74 (s, 3H) ppm; APCI MS mlz 540 [M+Ht. Example 38. Preparation of compound 67. Br 6-bromopyrazolo[ 4,3-b]pyridine 0 Hcf H 67 Prepared according General Procedure B from compound All (200 mg, 0.45 mmol) and 6- bromo-1H-pyrazolo[4,3-b]pyridine (450 mg, 2.27 mmol), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 67 as an off-white solid (43 mg, 17%): mp 102-105 °C; 1H NMR (300 MHz, CDCb) o 8.60 (d, J = 1.5 Hz, lH), 8.23 (s, IH), 7.75 (s, lH), 5.16 (d, JAF3 = 18.0 Hz, lH), 5.06 (d, JAB= 18.0 Hz, lH), 3.48 (d, J = 10.0 Hz, lH), 3.38 (d, J = 10.0 Hz, IH), 3.30 (s, 3H), 2.68 (t, J = 9.3 Hz, lH), 2.30- 1.93 (m, 3H), 1.92-1.38 (m, llH), 1.37--0.80 (m, 12H), 0.73 (s, 3H) ppm; ESI MS mlz 558 [M+Ht. Example 39. Preparation of compounds 68 and 69. 111 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 O Br . H Hd A A11 6-chloropyrazolo [4,3-b]pyridine K2C03, THF, rt PCT / US2014 / 052417 O :μCl ~ ~ I, N:,-._ -N H . HO H 69 Prepared according General Procedure B from compound All (86 mg, 0.19 mmol) and 6-chlorolH- pyrazolo[ 4,3-b ]pyridine (570 mg, 3.7 mmol), with purification by reverse phase preparative HPLC to provide 68 as a white solid (12 mg, 12%): mp 88-90 °C; 1H NMR (500 MHz, CDCh) 8 8.59 (s, lH), 8.51 (s, lH), 8.34 (s, lH), 5.40-5.22 (m, 2H), 3.48 (d, J = 10.0 Hz, lH), 3.38 (d, J = 10.0 Hz, IH), 3.30 (s, 3H), 2.67 (t, J = 8.6 Hz, lH), 2.27-2.21 (m, lH), 2.11-1.86 (m, 2H), 1.85- 1.41 (m, llH), 1.34-1.09 (m, llH), 1.04-0.93 (m, lH), 0.91-0.81 (m, lH), 0.73 (s, 3H) ppm; APCI MS mlz 514 [M+Ht. Further elution provided 69 as a white solid (40 mg, 40%): mp 115-117 °C; 1H Nl\1R (500 MHz, CDCh) 8 8.58 (s, lH), 8.40 (s, lH), 7.77 (s, lH), 5.18 (dd,J= 18.0, 32.1 Hz, 2H), 3.49 (d,J= 10.0 Hz, lH), 3.38 (d, J= 10.0 Hz, lH), 3.30 (s, 3H), 2.69 (t,J= 8.9 Hz, lH), 2.24-2.18 (m, lH), 2.14-2.10 (m, lH), 2.07-2.03 (m, lH), 1.80-1.68 (m, 4H), 1.67-1.56 (m, 3H), 1.55-1.42 (m, 4H), 1.39-1.11 (m, l0H), 1.05-0.98 (m, lH), 0.91-0.85 (m, lH), 0.73 (s, 3H) ppm; APCI MS mlz 514 [M+Ht. Example 40. Preparation of compounds 70, 71, and 72. 5-chlorobenzotriazole Prepared according General Procedure B from compound A21 (200 mg, 0.44 mmol) and 5- chlorobenzotriazole (1.35 g, 8.8 mmol), with purification by reverse phase preparative HPLC to 112 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 provide 72 as a white solid (12.6 mg, 5%): mp 76-77 °C; 1HNMR (500 MHz, CDCb) o 7.87 (d, .! = 1.5 Hz, lH), 7.81 (d, .!= 9.0 Hz, lH), 7.34 (dd, .!= 9.0, 1.5 Hz, lH), 5.50 (dd, J= 32.5, 17.0 Hz, 2H), 3.52 (d, .! = 9.5 Hz, lH), 3.43-3.38 (m, 3H), 2.65 (t, J = 9.0 Hz, lH), 2.25-2.23 (m, lH), 2.14-2.13 (m, lH), 2.06-2.04 (m, lH), 1.77-1.48 (m, lOH), 1.41-1.38 (m, IH), 1.33-1.11 (m, 13H), 1.02-1.00 (m, lH), 0.88-0.86 (m, lH), 0.75 (s, 3H) ppm; ESI MS mlz 528 [M+Ht. Fmther elution provided 70 as a white solid (17.1 mg, 7%): mp 69-70 °C; 1HNMR (500 MHz, CDCb) o 8.00 (d, .! = 9.0 Hz, lH), 7.35-7.33 (m, 2H), 5.37 (dd, .! = 40.5, 18.0 Hz, 2H), 3.53 (d, .! = 9.5 Hz, lH), 3.43-3.40 (m, 3H), 2.72 (t, J= 9.0 Hz, lH), 2.24-2.23 (m, lH), 2.16-2.14 (m, lH), 2.08-2.06 (m, lH), 1.79-1.42 (m, llH), 1.37-1.13 (m, llH), 1.01-0.99 (m, lH), 0.88 (t, J= 7.0 Hz, 3H), 0. 74 (s, 3H) ppm; ESI MS mlz 528 [M+Ht. Further elution provided 71 as a white solid (23.6 mg, 10%): mp 82-83 °C; 1HNMR (500 MHz, CDCb) o 8.06 (d, J= 1.5 Hz, lH), 7.45 (dd, .!= 9.0, 1.5 Hz, IH), 7.28 (d, J= 9.0 Hz, IH), 5.40 (dd, J = 29.5, 18.0 Hz, 2H), 3.53 (d, J = 10.0 Hz, lH), 3.42-3.39 (m, 3H), 2.71 (t, J = 9.0 Hz, IH), 2.24-2.23 (m, lH), 2.16-2.14 (m, lH), 2.08-2.05 (m, lH), 1.77-1.48 (m, llH), 1.33-1.13 (m, 1 lH), 1.02-0.99 (m, lH), 0.88 (t, J = 7.0 Hz, 3H), 0. 74 (s, 3H) ppm; ESI MS ml= 528 [M+Ht. Example 41. Preparation of compound 76. 0 Me~') r;ijBr 5-chlorobenzotriazole ~)A K2C03, THF, rt HCJ' H C11 Prepared according General Procedure E, Step 2 from compound Cll (120 mg, 0.27 mmol) and 5-chlorobenzimidazole (125 mg, 0.82 mmol), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 76 as a white solid (19 mg, 14%): mp 85-87 °C; 1H NMR (300 MHz, CDCh) 8 7.88-7.80 (m, 2H), 7.35 (d, J= 9.1 Hz, IH), 5.49 (s, 2H), 3.55 (d, J = 9.0 Hz, lH), 3.34 (s, 3H), 3.22 (d, J = 9.0 Hz, IH), 2.64 (t, J = 8.5 Hz, lH), 2.27-2.12 (m, 2H), 1.94-1.85 (m, 2H), 1.84-1.36 (m, 15H), 1.35-1.14 (m, lOH), 0.73 (s, 3H) ppm; APCI MS m / z 514 [M+Ht. 113 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 Example 42. Preparation of compounds 77, 78, and 79. 4,5-difluorobenzotriazole PCT / US2014 / 052417 Prepared according General Procedure E, Step 2 from compound Cll (120 mg, 0.27 mmol) and 4,5-difluoro-1H-benzo[d][l,2,3]triazole (126 mg, 0.82 mmol), with semi-purification by column chromatography on silica gel followed by reverse phase preparative HPLC to provide 79 as a white solid (62 mg, 44%): mp 103-105 °C; 1H NMR (500 MHz, CDCh) & 7.64 (ddd, J = 9.0, 3.5, 1.0 Hz, IH), 7.32-7.26 (m, lH), 5.54 (d, .fl\.B = 17.0 Hz, lH), 5.49 (d, JAB= 17.0 Hz, lH), 3.54 (d, J = 9.0 Hz, IH), 3.34 (s, 3H), 3.22 (d, J = 9.0 Hz, lH), 2.65 (t, J = 9.0 Hz, l H), 2.27- 2.18 (m, lH), 2.17-2.11 (m, lH), 1.97-1.88 (m, 2H), 1.84-1.73 (m, 3H), 1.68-1.40 (m, 9H), 1.40-1.11 (m, lOH), 0.74 (s, 3H) ppm; ESI MS mlz 514 [M-Hr. Further elution provided 77 as a white solid (20 mg, 14%): mp 98-100 °C; 1H NMR (500 MHz, CDCh) & 7.81 (ddd, J = 9.0, 4.0, 1.0 Hz, lH), 7.24-7.19 (m, lH), 5.54 (d, JAB= 18.5 Hz, lH), 5.50 (d, JAB= 18.5 Hz, lH), 3.55 (d, J = 9.0 Hz, IH), 3.34 (s, 3H), 3.22 (d, J = 9.0 Hz, IH), 2.71 (t, J= 9.0 Hz, lH), 2.27-2.17 (m, lH), 2.17-2.10 (m, lH), 1.98-1.89 (m, 2H), 1.88-1.72 (m, 3H), 1.70-1.42 (m, 9H), 1.40-1.10 (m, l0H), 0.72 (s, 3H) ppm; ESI MS m / z 514 [M-Hr. Further elution provided 78 as an off-white solid (39 mg, 28%): mp 100-102 °C; 1H NMR (500 MHz, CDCh) 8 7.41-7.34 (m, lH), 7.05 (ddd, J = 9.0, 3.0, 1.0 Hz, IH), 5.44 (d, JAB= 18.0 Hz, IH), 5.36 (d, . / 1\.B = 18.0 Hz, lH), 3.53 (d, J = 9.0 Hz, lH), 3.34 (s, 3H), 3.23 (d, J = 9.0 Hz, lH), 2.70 (t, J = 9.0 Hz, lH), 2.26-2.17 (m, lH), 2.17-2.11 (m, lH), 1.98-1.88 (m, 2H), 1.84-1.73 (m, 3H), 1.69-1.42 (m, 9H), 1.41-1.12 (m, l0H), 0.71 (s, 3H) ppm; ESI MS m / z 514 [M-Hr. Example 43. Preparation of compound 80. 114 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Br benzimidazole Prepared according General Procedure E, Step 2 from compound Cll ( 100 mg, 0.23 rnmol) and benzimidazole (80 mg, 0.68 mmol), with purification by reverse phase preparative HPLC to provide 80 as a white solid (68 mg, 63%): mp 130-132 °C; 1H NMR (300 MHz, CDCh) 8 8.74 (b s, lH), 7.91-7.87 (m, lH), 7.41-7.38 (m, 2H), 7.25-7.22 (m, lH), 5.16 (dd, J= 18.3, 5.8 Hz, 2H), 3.54 (d, J = 9.1 Hz, lH), 3.34 (s, 3H), 3.23 (d, J = 9.1 Hz, lH), 2.71 (t, J = 8.9 Hz, lH), 2.29- 2.08 (m, 2H), 1.94-1.09 (m, 26H), 0.72 (s, 3H) ppm; APCI MS mlz 479 [M+Hf. Example 44. Preparation of compounds 81, 82, and 83. MeO'> rirBr ~ HO H C11 4-fluorobenzotriazole Prepared according General Procedure E, Step 2 from compound Cll (120 mg, 0.27 mmol) and 4-fluoro-1H-benzo[d][l,2,3]triazole (112 mg, 0.82 mmol), with purification by reverse phase preparative HPLC to provide 82 as an off-white solid (40 mg, 30%): mp 199-201 °C; 1H NMR (500 IvlHz, CDCh) 8 7.45-7.40 (m, lH), 7.11 (d, J = 8.3 Hz, lH), 7.06-7.02 (m, lH), 5.41 (dd, J = 18.0, 9.2 Hz, 2H), 3.54 (d, J = 9.0 Hz, lH), 3.34 (s, 3H), 3.23 (d, J = 9.0 Hz, lH), 2. 70 (t, J = 8.9 Hz, IH), 2.27-2.13 (m, 2H), 1.97-1.89 (m, 2H), 1.83-1.68 (m, 5H), 1.67-1.43 (m, 9H), 1.41- 1.12 (m, 9H), 0. 72 (s, 3H) ppm; APCI MS mlz 498 [M+Hf. 115 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Further elution provided 81 as a white solid (20 mg, 15%): mp 98-100 °C; 1H NMR (500 MHz, CDCh) o 7.86 (d, J = 8.4 Hz, lH), 7.31-7.26 (m, IH), 7.15-7.11 (m, lH), 5.43 (dd, J = 18.2, 0.95 Hz, 2H), 3.55 (d, J = 9.0 Hz, IH), 3.34 (s, 3H), 3.22 (d, J = 9.0 Hz, lH), 2.70 (t, J = 9.0 Hz, lH), 2.25-2.12 (m, 2H), 1.97-1.75 (m, 6H), 1.70-1.44 (m, 9H), 1.39-1.13 (m, IOH), 0.72 (s, 3H) ppm; APCI MS mlz 498 [M+Ht, Fmther elution provided 83 as an off-white solid (48 mg, 36%): mp 172-174 °C; 1H NMR (500 MHz, CDCh) o 7.67 (d, J = 8.6 Hz, lH), 7.35-7.30 (m, lH), 7.06-7.02 (m, lH), 5.53 (dd, J = 17.0, 2.6 Hz, 2H), 3.55 (d, J= 9.0 Hz, lH), 3.34 (s, 3H), 3.21 (d, J= 9.0 Hz, IH), 2.65 (t, J= 9.0 Hz, lH), 2.26-2.13 (m, 3H), 1.96-1.89 (m, 2H), 1.83-1.71 (m, 3H), 1.67-1.40 (m, 9H), 1.37-1.11 (m, 9H), 0.74 (s, 3H) ppm; APCI MS mlz 498 [M+Hr. Example 45. Preparation of Compound 4. Br 0 H Hc5 A () 0 A11 4 To a solution of compound All (40 mg, 0.09 mmol) in THF (2 mL) was added morpholine (390 mg, 4.5 m:mol) and K2CO3 (120 mg, 0.9 mmol). The resulting solution was stirred at room temperature overnight. Then LCMS showed the reaction was complete. The reaction was diluted with EtOAc (40 mL), and washed with brine (15 mL x 2). The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by prep-HPLC to give compound 4 (18 mg, 45%) as a white solid. Compound 4: 1H NMR: (500 MHz, CDCh), o (ppm), 3.79-3.77 (m, 4H), 3.48 (AB, IH, J=l0 Hz), 3.38 (AB, IH, J=IO Hz), 3.30 (s, 3H), 3.22 (s, 2H), 2.58 (t, lH, J=9.3 Hz), 2.5 (s, 4H), 1.25 (s, 3H), 0.66 (s, 3H). Example 46. Preparation of Compound 2. 116 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 Br A11 K2C03, THF ~ CN PCT / US2014 / 052417 2 To a solution of compound All (40 mg, 0.09 mmol) in THF (2 mL) was added IH-pyrazole (300 mg, 4.5 mmol) and K2CO3 (120 mg, 0.9 mmol). The resulting solution was stirred at room temperature overnight. Then LCMS showed the reaction was complete. The reaction was diluted with EtOAc (40 mL), and washed with brine (15 mL x 3). The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by prep-HPLC to give 2 (16 mg, 40%) as a white solid. Compound 2: 1H NMR: (500 MHz, CDCb), 8 (ppm), 7.57 (d, IH, J=l Hz), 7.43 (d, lH, J=l.5 Hz), 6.35 (s, lH), 4.98 (AB, lH, J=l 7.5 Hz), 4.90 (AB, lH, J=18 Hz), 3.48 (AB, IH, J=I0.5 Hz), 3.39 (AB, IH, J=9.5 Hz), 3.31 (s, 3H), 2.60 (t, IH, J=S.8 Hz), 1.25 (s, 3H), 0.72 (s, 3H). Example 47. Preparation of Compound 5. Br K2C03, THF H HCf H () N Hd H A11 .,):,0 5 Compom1d All (30mg, 0.07mmol), K2CO3 (50mg) and l-(piperazin-1-yl)ethanone (200mg) were dissolved in THF (3 mL) and stirred at room temperature overnight. The solvent was removed in vacuo and the residue was purified by prep-HPLC to give compound 5 (6 mg, 20%) as a white solid. Compound 5: 1H NMR: (400 MHz, CDCh), 8 (ppm), 3.68-3.66 (m, 2H), 3.51 (t, 2H, J=5 Hz), 3.45 (AB, lH, J=l0 Hz), 3.37 (AB, IH, J=l0 Hz), 3.28 (s, 3H), 3.21 (s, 2H), 2.52-2.44 (m, 5H), 2.08 (s, 3H), 1.23 (s, 3H), 0.64 (s, 3H). 117 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 Example 48. Preparation of Compound Bll. HO')r;K 0~ 81 DHP.PPTS THF Me3SOI ff.OH ~NaH. DMSO H o' i:i B4 EtPPh3Br t-BuOK, THF H3C Hc5 H B7 0 PTSA MeOH H3C H3C Hc5 H Hc5 B9 Li, liq.NH3 t-BuOH, THF LiAIH4 THF OH ~ HO A B5 1). BH3_THF 2).NaOH, H202 88 0 Br2, HBr MeOH H3C H Hc5 810 PCT / US2014 / 052417 ~ PO,", OH H H 0 :. H 83 0 ~ HO A B6 OH PCC CH2Cl2 Br 0 H 811 Step 1. Preparation of Compound B2. Compound Bl (10.0g, 33mmol) was dissolved in 100 mL ofTHF. Dihydropyran (25ml, 270mmol) and PPTS (4.16, l6mmol) was added and the resultant reaction mixture was vigorously stiffed for 15 hat room temperature. Upon concentration under reduced pressure ,the reaction rnixture was taken up in EtOAc (500 mL) , washed with water (300 mL) and brine (300 mL), dried over sodium snlfrtte and concentrated under reduced pressure, The residue vvas purified by chromatography on silica gel ( eluant: petroleum ether / EtOAc =lOil--3 / 1) to afford compound B2 12.52 g (97.65~,'i,). Compound B2: LC-MS: m / z=409.0 [M +Nat 118 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Step 2. Preparation of Compound B3. Lithium metal (3.0 g, 0.4 mmol) were added to condensed ammonia (500 ml) in a three neck flask at-70°C. Then a solution of Compound B2 (5.0 g, 13mmol) and tert-BuOH (0.95 g, 13mmol) in anhydrous tetrahydrofuran (100 mL) was added dropwise and stirred for 0.8 hours. Ammonium chloride (30.0 g) was added to quench the reaction and the ammonia was left to evaporate overnight. The residue was extracted with EtOAc (300 mL). The organic layers were washed with saturated NaCl solution (2x200 mL), dried over Na2SO4 and concentrated under reduced pressure, The residue was purified by chromatography on silica gel (eluant: petroleum ether / EtOAc =10 / 1-2 / 1) to afford 2.0 g of compound 3 (39.60%). Compound B3: LC-MS: m / z=413.3 [M +Nar Step 3. Preparation of Compound B4. Me3SOI (16.9g,76.80mmol) was dissolved in 80 mL of DMSO and NaH (1.84g, 76.80mmol) was added. The mixture was stirred at room temperature for 1 hom, then compound B.3 (6.0 g, 15.36 mmol) dissolved in 60 mL ofDMSO was added. The solution was stirred at room temperature overnight. Water ( 10 mL) was then added to the reaction mixture. The aqueous reaction mixture was extracted with EtOAc (300mL x 3). The extracts were dried over Na2SO4, filtered, concentrated. The cmde compound B4 was directly used in the next step without further purification. Step 4. Preparation of Compound BS. The crude compound B4 was slowly added into a suspension of LiA1H4 ( 1. 75 g, 51 mmol) in l 00ml of dry THF at O °C. The mixture was stirred at room temperature for 2h, then 2. lg of 15% aq NaOH was slowly added to quench the reaction. The reaction mixture extracted with EtOAc (200mL x 3). The organic layers were dried over MgSO4, filtered, and concentrated. The crude compound B5 was directly used in the next step without further purification. Step 5. Preparation of Compound B6. The crnde compound B5 was dissolved in 100ml of dry CH2C}z, and 4.0 g of PCC was added at O °C. Then the mixtme was stirred at room temp for 6h. The reaction mixture was then filtered, concentrated, and purified by flash chromatography on silica gel using 10 / 1~3 / l petroleum ether: ethyl acetate=l0 / l-3il elution to give compound B6, 3.10 g (50.89%, three-step yield). Step 6. Preparation of Compound B7. To a suspension of Ethyltriphenylphosphonium bromide (14.20g, 38.3mmol) in dry THF (40mL) was added KOtBu (4.30 g, 38.3mmol) under 119 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 N2 atmosphere. The mixture was heated at reflux forl hour, during which time the mixture turned bright orange. Then compound B6 (3.1 g, 7.66mmol) in dry THF (25mL) was added to the above refluxing solution and stirred at reflux overnight. After cooling to room temperature, the solution \Vas poured into brine (l00mL). The aqueous solution was extracted with ethyl acetate ( 1 00mL x 3 ). The extracts were washed with brine (30mL x 2 ), dried over Na2SO4 ,filtered ,concentrated and purified by column chromatography on silica gel (petroleum ether / EtOAC from 10 / 1 to 4 / 1) to give compound B7 2.2g (68.97%) as white solid. Furthermore, the C-3 isomer (0.30 g, 9.63%) was also obtained. Step 7. Preparation of Compound B8. To a solution of compound B7 (3 g, 7.2mmol) in dry THF (20mL) was added borane-tetrahydrofuran complex (29mL of 1.0 M solution in THF) and the reaction mixture was stirred at ambient temperature for 1 hour. 10% aqueous NaOH (20 mL) was slowly added. The mixture was cooled in ice and 30% aqueous solution of H202 (20mL) was slowly added. The mixture was stirred at ambient temperature for 1 hour and then extracted with CH2Ch (3 x 100 mL). The combined CH2Ch extracts were washed with 10% aqueous Na2S203 (50 mL), which was directly used in the next step without further purification. Step 8. Preparation of Compound B9. The combined CH2Ch extracts of the compound B8 of last step was used without further purification. 3.5g of PCC was added at 0 °C. Then the mixture was stirred at room temperature for 6h , The mixture was filtered, concentrated, and purified by flash chromatography on silica gel using 12 / 1-7 / l(petroleum ether :ethyl acetate) elution to give 1.28 g of compound B9 (41.23% two steps). Compound B9: LC-MS: m / z=455.3[M +Na]+. 1H NMR (500 MHz, CDCI3) o(ppm): 4.57&4.53(1H,t,J=3.5Hz), 3.96&3.87(1H,AB, J=l l.0Hz,),3.82 (IH,t, J=9.5Hz), 3.56-3.53(1H,m), 3.44&3.27 (IH,AB, J=10.5Hz), 2.53(1H,t, J=9.0Hz), 2.12&2.11(3H,s), 1.22&1.21(3H,s), 0.64&0.61(1H,s). Step 9. Preparation of Compound BlO. Compound B9 (1.28g, 2.96mmol) was dissolved in 50mL of dry MeOH and I 00 mg of PTSA was added. The reaction mixture was stirred at room temperature overnight. The reaction mixture was then concentrated under reduced pressure. This product mixture was separated by flash chromatography on silica gel using 8 / 1 ~ 2 / 1 (petroleum ether:EtOAc) elution to give 674 mg of compound BlO (65.32%). Compound BlO: LC-MS: m / z=331.3[M-H2O+H]\ m / z=349.2[M +Ht 1H NMR (500 MHz, CDC13) o(ppm): 120 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 3.89 (lH, AB, J=l2.0 Hz), 3.72 (lH, AB, J=l2.0 Hz), 2.53 (lH, t, 1=9.0 Hz), 2.11 (3H, s), 1.22 (3H, s), 0.64 (3H, s). 13C NMR (125.77 MHz, CDC13) 8(ppm): 209.78, 69.68, 63.80, 60.12, 57.07, 54.39, 44.36, 42.04, 41.18, 39.62, 39.31, 36.05, 35.39, 31.85, 31.68, 31.54,28.04, 27.91, 24.39, 22.86, 22.75, 13.72. Step 10. Preparation of Compound B11. Compound BIO (50 mg, 0.14 mmol) was dissolved in 5 mL of dry MeOH and 3 drops of Br2 and 2 drops of HBr aq. was added. The reaction mixture was stirred at room temperature for 3h, then the reaction mixture was treated with triethylamine at O °C, concentrated under reduced pressure and was directly used in the next step without further purification. Compound BU: LC-MS: m / z=410.1&41 l.2[M-H2O+H(. Example 49. Preparation of Compound 9. Br B11 Co) N H 9 Cmde compound Bll was directly used, 8 mL THF and 100mg of K2CO3 , 0.5ml of morpholine was added. The reaction mixture was stirred at room temperature overnight. The solution was diluted with ethyl acetate (100 mL). The resulting solution was washed with brine (100 mL), dried over sodium sulfate and concentrated in vacuo. The residue was purified with reverse phase prep-HPLC to give 25 mg ( 41.18%, two steps from 50mg of compound B10) product 9 as white solid. Compound 9: 1H NJ\t1R (500 MHz, CDC13) 8(ppm): 3.89(1H,dd, J=4.0Hz, J=l 1.5Hz), 3.76 (4H,t, J=4.5Hz), 3.72 (IH,dd, J=3.0Hz, J=l l.0Hz), 3.19 (2H,s), 2.58 (lH,t, J=9.5Hz), 2.45-2.55 (4H, m), 2.20-2.IS(lH,m), 2.07-2.04(1H,m), l.92~1.89(1H,m),1.23(3H,s), 0.67(3H, s). Example 50. Preparation of Compounds 12 & 10. 121 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 Br 811 12 N',N) \ !. N + PCT / US2014 / 052417 10 N0--._ I / / N-N Crude compound Bl 1 was directly used, and 8 mL of dry THF and 100mg of K2CO3 ,0.5ml of I H-1,2,3-triazole was added. The reaction mixture was stirred at room temperature overnight. The solution was diluted with EtOAc (100 mL). The resulting solution was washed with brine ( 100 mL), dried over sodium sulfate and concentrated in vacuo. The residue was purified with reverse phase prep-HPLC to give 10 mg of Compound 12 and 19 mg of Compmmd 10 as white solid. Compound 12: 1H NMR(500 MHz, CDC13) 8(ppm): 7.76(1H,s), 7.64(1H, s), 5.24 (lH,AB, J=l7.5Hz), 5.16 (IH,AB, J=18.0Hz),3.89 (IH,AB, J=l 1.5Hz), 3.73 (IH,AB, J=l 1.5Hz),2.65 (lH,t, J=9.0Hz), 2.25~2.19(1H,m), 2.10~2.05(2H,m), l.23(3H,s), 0.71(3H, s). Compound 10: 1H NMR (500 MHz, CDC13) 8(ppm): 7.68 (2H,s), 5.25(1H,AB,J=17.5Hz), 5.22(IH,AB, J=l 7.5Hz)3.89 (IH,AB, J=l 1.5Hz), 3.73 (IH,AB, J=l 1.5Hz), 2.58 (IH,t, J=8.5Hz), 2.24"-2.20(1H,rn), 2.11~2.04(21H,m), l.23(3H,s), 0.75(3H, s), Example 51. Preparation of Compound 11. n Br N-N ► B11 11 Crude compound Bl 1 was directly used, 8 mL of dry THF and I 00mg of K2CO3 , 0 .5rnl of pyrazole was added. The reaction mixture was stirred at room temperature overnight. The solution was diluted with ethyl acetate (100 mL). The resulting solution was washed with brine ( 100 mL), dried over sodium sulfate and concentrated in vacuo. The residue was purified with reverse phase prep-HP LC to give 30 mg of Compound 11 white solid. Compound 11: 1H NMR 122 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 (500 MHz, CDC13) 8(ppm): 7.54 (lH,d, J=l.0Hz), 7.40 (lH,dt, J=2.0Hz), 6.33 (lH,t, J=l.5Hz), 4.94(1H,AB, J=l 7.5 Hz), 4.90(1H,AB, J=l7.0 Hz), 3.88(1H,AB, J=l 1.5 Hz), 3.73(1H,AB, J=l2.0 Hz), 2.58 (lH,t, J=8.5Hz) 2.23-2.17 (1H,m),2.07-2.05(2H,m), 1.23(3H,s), 0.72 (3H, s). Example 52. Preparation of Compound 6. 811 6 Crude compound Bll was directly used, 8 mL of dry THF and 100mg ofK2CO3 ,0.5ml of l( piperazin-1-yl)ethanone was added. The reaction mixture was stirred at room temperature overnight. The solution was then diluted with ethyl acetate (100 mL). The resulting solution was washed with brine (100 mL), dried over sodium sulfate and concentrated in vacuo. The residue was purified with reverse phase prep-HPLC to give 29 mg (43.64%,t wo steps from 50mg of compound B10) Compound 6 as white solid. Compound 6: 1H NMR (500 MHz, CDC13) o(ppm): 3.89(1H,AB, J=l 1.5 Hz), 3.72(1H,AB, J=l 1.5 Hz), 3.68~3.65 (2H,m),3.51(2H,t, J=5.0Hz) 3.21(3H,s), 2.55 (IH,t, J=9.5Hz), 2.45 (3H, t, J=5.0Hz), 1.23 (3H, s), 0.67(3H, s) Example 53. Preparation of Compound 7. 811 7 59mg (0J2mmol) of crude Compound BU was dissolved in 8 mL THF and 100mg (0.77mmol) of K2CO3, 100mg (0.6lmrnol) of 1-(Methylsulfonyl)piperazine was added. The reaction mixture 123 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 was stirred at room temperature overnight. The solution was diluted with ethyl acetate (100 mL). The resulting solution was washed with brine ( 100 mL ), dried over sodium sulfate and concentrated in vacuo. The residue was purified with reverse phase prep-HPLC to give 8 mg (0.02 rnmol,10.9%) product 7 as white solid. Compound 7: 1H NMR (500 MHz, CDC13) 8(ppm): 3.89(1H,AB, J=l 1.5Hz), 3.72(1H,AB, J=l2Hz), 3.30(4H,t, J=5.0Hz), 3.25(2H, s), 2.78(3H, s) 2.61(4H,t, J=5.0Hz), 2.52(1H,t, J=8.5Hz), 2.13~2.0l(lH,m), 1.23(3H,s), 0.67(3H, s). Example 54. Preparation of Compound E15. rt, overnight E1 1) Pd / C, H2, EA 2) HCl(7.3%) E4 0 E2 pyridine.HCI glycol, toluene LiOH NaH, Mel THF / MeOH 0 o A E6 K-oolectrtde, THF 6) ~ HO''.~ H E9 Date Rec;ue / Date Received 2024-04-12 E7 PPhaCH2CH38r t-BuOK, THF 124 0 CH(OEt)3, p-TsOH ~ 1,4-dioxane, EtOH, rt, 5h EtO E3 E5 b, ~-o aq.HCI ~ 0 - H ES E10 WO 2015 / 027227 PCT / US2014 / 052417 imidazole ~ TBSCI H __ D_M_F _.TBSO''' - - H 9-BBN TBSO'' OH E11 E12 0 Br 0 Dess-Martin TFA TBSO'' HO'' E13 E14 E15 Step 1. Preparation of E2. To a solution of El (250.0 g, 0.83 mol) in pyridine (1 L) was added Ac2O (168.8 g, 1.65 mol) dropwise at 19°C. After the addition was completed, the reaction mixture was stirred at l 9°C overnight. TLC (petroleum ether: ethyl acetate =I: l) showed that the reaction was completed. Then the reaction mixture was concentrated in vacuum, the residue was poured to water and extracted with dichloromethane (3x500 mL), the organic layers were washed with 2 N HCl (200 mL), saturated NaHCO3(300 mL), brine and dried over anhydrous sodium sulfate, filtered, concentrated to give the crude target product E2 (283.7 g, 99.7%) as brown oil. 1H NMR (E2): (400 MHz, CDC13) o 5.95-5.90 (m, lH), 4.70-4.64 (m, IH), 4.20-4.13 (m, lH), 2.68-2.55 (m, IH), 2.54-2.30 (m, 5H), 2.15-1.90 (m, 6H), 1.90-1.75 (m, 4H), 1.63-1.38 (m, 2H), l.33-1.07 (m, 4H), 0.90 (s, 3H). Step 2. Preparation of E3. To a solution of E2 (250.0 g, 0.73 mol) in 1,4-dioxane (700 mL) and EtOH (467 mL) was added CH(OEt)3 (227.2 g, 1.53 mol) and p-TsOH (2.8 g, 14.60 mmol) at 29°C. After the addition was completed, the reaction mixture was stirred for 1 hr at 29°C. TLC (petroleum ether: ethyl acetate =3:1) showed that the reaction was completed. Then the reaction mixture was quenched with saturated NaHCO3 (300 mL) and poured to water, extracted with EtOAc (3x500 mL), the organic layers were washed with brine and dried over anhydrous sodium sulfate, filtered, concentrated, recrystallized from petroleum ether: ethyl acetate= 10: 1 to give the target product E3 (155.8 g, 57.6%) as a white solid. 1H NMR (E3): (400 MHz, CDC13) 8 5.42-5.38 (m, IH), 5.15-5.10 (m, IH), 4.46-4.50 (m, IH), 4.32-4.25 (m, lH), 4 .. 05- 125 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 3.98 (m, lH), 3.85-3.70 (m, 2H), 2.53-2.42 (m, lH), 2.37-2.20 (m, 3H), 2.18-1.91 (111, 7H), 1.90- 1.72 (m, 3H), 1.62-1.48 (m, 2H), 1.40-1.20 (m, 6H), 1.20-1.12 (m, IH), 0.90 (s, 3H). Step 3. Preparation of E4. To a solution of E3 (30.0 g, 80.54 mmol) in EtOAc ( 400 mL) was added Pd / C ( 1.5 g, 50% water) under N2. The reaction mixture was degassed under vacuum and purged with H2 several times. Then the reaction mixture was stirred for 1 hr at l 5°C under H2 atomsphere. Then it was filtered and the filtrate was stirred at 15°C, 10% HCl (100 mL) was added and the reaction mixture was stirred for 1 hr at 15°C. TLC (petroleum ether: ethyl acetate =3:1) showed that the reaction was completed. The reaction mixture was poured to water and extracted with EtOAc (3x200 mL), the organic layers were washed with saturated NaHCO3, brine and dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column (petroleum ether: ethyl acetate=l0: 1-3: 1) to give the product E4 (33.0 g, yield: 59.1 %) as colorless oil. 1H NMR (E4): (400 MHz, CDCI3) S 4.75-4.50 (m, lH), 4.45-4.38 (m, lH), 2.55-2.35 (m, SH), 2.31-2.20 (m, lH), 2.15-2.05 (m, 4H), 2.05-1.65 (m, 6H), 1.60-1.15 (m, 7H), 1.15-1.00 (m, IH), 0.95-0.85 (m, 4H). Step 4. Preparation of ES. To a solution ofE4 (31.0 g, 89.48 mmol) and ethane-1,2-diol (50 mL) in toluene (200 mL) was added cat. amount of pyridine.HCl (0.3 g, 2.60 mmol) at 16°C. After the addition was completed, the reaction mixture was heated to reflux and removed water by Dean-Stark trap for 18 hr. TLC (petroleum ether: ethyl acetate =3:1) showed that the reaction was completed. Then the reaction mixture was cooled to 16°C, poured to water, and extracted with EtOAc (3xl00 mL), the organic layers were washed with brine and dried over anhydrous sodium sulfate, filtered, concentrated to give the crude product E5 (36.9 g, yield: 94.8 %) as a white solid., which was used for next step. 1H NMR (E5): (400 MHz, CDCl3) 5 4.40-4.30 (m, 2H), 4.20-4.10 (m, 2H), 4.00-3.80 (m, 8H), 2.20-1.90 (m, SH), 1.85-1.05 (m, 20H), 1.05-0.75 (m, SH). Step 5. Preparation of E6. To a solution of ES (55.0 g, 126.56 mmol) in THF (200 mL) and MeOH (50 mL) was added 4 N LiOH (94.9 mL, 379.69 mmol) at 20°C. After the addition was completed, the reaction mixture was stirred for 18 hr at 20°C. TLC (petroleum ether: ethyl acetate =3: 1) showed that the reaction was completed. Then the reaction mixture was poured to water, and extracted with EtOAc (3x200 mL), the organic layers were washed with brine and 126 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 dried over anhydrous sodium sulfate, filtered, concentrated to give the crude product E6 ( 46.3 g, yield: 93.1 %) as a white solid. 1H NMR (E6): (400 l\!IHz, CDC13) 8 4.00-3.75 (m, lOH), 2.25- 2.15 (m, lH), 2.05-1.90 (m, lH), 1.85-1.60 (m, SH), 1.60-1.40 (m, 7H), 1.35-1.00 (m, 5H), 1.00- 0.75 (rn, 5H). Step 6. Preparation of E7. To a suspension of 60% NaH (9.6 g, 0.24 rnol) in dry THF (100 rnL) was added dropwise a solution of compound E6 (46.3 g, 0.12 mol) in dry THF (200 mL) at 25°C under N2. The mixture was stirred at for 30 min, then Mel (51.1 g, 0.36 mol) was added dropwise at 25°C. After the addition was completed, the reaction mixture was stirred for 4 hr at 45 °C. TLC (petroleum ether: ethyl acetate =3: 1) showed that the reaction was completed. The reaction was cooled to room temperature, quenched with saturated NH4Cl (200 mL), poured to water, extracted with EtOAc (3x200 mL), the organic layers were washed with brine and dried over anhydrous sodium sulfate, filtered, concentrated to give the crude product E7 (50.0 g, crude) as yellow solid. 1H NMR (E7): (400 MHz, CDC13) 8 3.95-3.80 (m, SH), 3.55-3.40 (m, 2H), 3.28 (s, 3H), 2.18-2.10 (m, IH), 2.01-1.95 (m, lH), 1.81-1.57 (m, SH), 1.46-1.37 (m, SH), 1.29-1.18 (m, 4H), 1.02-0.85 (m, 7H). Step 7. Preparation of ES. To a solution of compound E7 (50.0 g, 0.12 rnol) in THF (200 mL) and acetone (40 mL) was added aqueous 2 N HCl (40 mL). After the addition was completed, the reaction mixture was stirred for 18 hr at 25°C. TLC (petroleum ether: ethyl acetate =3:1) indicated the reaction was completed. Then the reaction mixture was poured to water, extracted with EtOAc (3x200 mL), the organic layers were washed with saturated NaHCO3, brine and dried over anhydrous sodium sulfate, filtered, concentrated to give the crude product ES ( 41.0 g, crude) as a yellow solid which was used to the next step directly. 1H NMR (ES): (400 MHz, CDC13) 8 3.73-3.58 (m, 2H), 3.34 (s, 3H), 2.55-2.40 (m, 3H), 2.42-2.30 (m, lH), 2.20-1.65 (rn, 8H), 1.60-1.20 (m, 1 OH), 1.10-0. 75 (m, 7H). Step 8. Preparation of E9. To a solution ofE8 (40.0 g, 125.7 mrnol) in dry THF (500 mL) was added dropwise K-selectride ( 151 mL, 150.8 mmol, 1 M in THF) at -78°C under N2. After the addition was completed, the reaction mixture was stirred for 3 hr at -78°C. TLC (petroleum ether: ethyl acetate =3:1) indicated the reaction was completed. The reaction mixture was slowly quenched with 30% H2O2 ( 17. l g, 150.5 mmol) at -78 °C, then reaction mixture was poured to 127 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 saturate NH4Cl, extracted with EtOAc (3xl 00 mL). The organic phase was washed with saturated Na2S2O3, brine, dried over sodium sulfate and evaporated to give the crude product. The crude product was purified by washing with PE: EtOAc= 10: 1 to give the target product E9 (23.5 g, 58%) as a white solid. 1H NMR (E9): (400 MHz, CDC13) o 4.10 (m, IH), 3.52 (d, 1H), 3.42 (d, IH), 3.30 (s, 3H), 2.50-2.40 (m, lH), 2.16-2.03 (m, lH), 1.98-1.90 (m, 2H), 1.86-1.64 (m, 10H), 1.55-1.47 (m, 3H), 1.32-1.17 (m, 6H), 1.12-0.95 (m, lH), 0.88-0.81 (m, 4H). Step 9. Preparation of ElO. To a suspension of Ph3PEtBr (25.97 g, 70 mmol) in dry THF (100 mL) was added dropwise a solution of t-BuOK (7.70 g, 70 mmol) in dry THF (50 mL) under N2 at 0°C. The mixture was stirred at room temperature for 1.5 h. Then a solution of E9 (2.8 g, 8.75 mmol) in THF (30 mL) was added dropwise and the resulting mixture was stirred at 60°C for 12 h. TLC (petroleum ether: ethyl acetate = 3: 1) indicated that the starting material was consumed completely. The reaction was quenched with saturated aqueous NH4Cl solution (100 mL) and exn·acted with EtOAc (50 mL x 2). The combined organic phases were dried over Na2SO4 and concentrated in vacuum. The residue was purified by column chromatography on silica gel (eluent: petroleum ether: ethyl acetate= 40:l) to give ElO (1.8 g, 62%) as white powder. 1H NMR (ElO) : (400 MHz, CDC13) o 5.15-5.05 (m, lH), 4.13-4.05 (m, lH), 3.55-3.35 (m, 2H), 3.30 (s, 3H), 2.45-1.90 (m, 4H), 1.80-1.35 (m, 13H), 1.30-0.95 (m, 8H), 0.90 (s, 3H), 0.85-0.70 (m, IH). Step 10. Preparation of Ell. To a solution ofElO (1.8 g, 5.41 mmol) in DMF (20 mL) was added imidazole (737 mg, 10.82 mmol) and TBSCl ( 1.22 g, 8.12 mmol). The mixture was stirred overnight at room temperature. TLC (petroleum ether: ethyl acetate = 3: 1) showed the starting material was consumed completely. The reaction was diluted with EtOAc (20 mL) and washed with brine (20 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The cmde product was purified by silica gel chromatography on silica gel eluted with petroleum ether to give Ell (2.33 g, 96%) as white solid. 1H NMR (Ell) : (400 MHz, CDC13) o 5.11-5.09 (m, lH), 4.01-4.00 (m, lH), 3.50-3.39 (m, 2H), 3.29 (s, 3H), 2.40-2.30 (rn, IH), 2.25-2.10 (rn, 2H), 1.90-1.82 (m, lH), 1.53-1.40 (rn, SH), 1.30-0.95 (m, 8H), 0.89-0.88 (m, 9H), 0.86-0.75 (m, 2H), 0.02-0.01 (m, 6H). 128 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Step 11. Preparation of E12. To a solution of 9-BBN (81 mL, 40.84 mmol) was added Ell (2.33 g, 5.06 mmol) in THF (20 mL). The mixture was stirred at 60°C for 16 h. Then the mixture was cooled to room temperature and added 10% NaOH aqueous (40 mL) and H2O2 (20 mL) dropwise. After stirred for 1 h, the mixture was quenched with aqueous Na2S2O3 and extracted with EtOAc ( I 00 mL). The organic layers combined and dried over anhydrous sodium sulfate. The organic phase was concentrated under vacuum. The crude product was purified by colunm chromatography on silica gel ( eluent: petroleum ether: ethyl acetate = 40: 1) to give E12 (2.3 g, 90%) as white solid. 1H NMR (E12) : (400 MHz, CDC13) 8 4.00-3.99 (m, IH), 3.71-3.65 (m, IH), 3.49-3.38 (m, 2H), 3.28 (s, 3H), 2.42-2.39 (m, lH), 1.89-1.82 (m, SH), 1.75-1.46 (m, 17H), 1.38-1.00 (m, 14H), 0.89 (s, 9H), 0.68 (s, 3H), 0.00 (m, 6H). Step 12. Preparation of E13. To a solution of E12 (2.1 g, 4.52 mmol) in CH2Ch (20 mL) was added Dess-Martin (3.8 g, 9.04 mmol). The mixture was stirred at 12°C for 5 h. TLC (petrolemn ether : ethyl acetate=3: I) showed the starting material was consumed completely. The mixture was quenched with a mixture solution ofNa2S2O3 / NaHCO3 (3:1, 12 g) in water (50 mL). The mixture was extracted with EtOAc (200 mL). The organic layers was dried over anhydrous sodium sulfate. The organic phase was concentrated under vacuum to afford E13 as white solid (2.3 g, crude). 1H NMR (E13) : (400 MHz, CDC13) 4.00-3.99 (m, lH), 3.49-3.38 (m, 2H), 3.27 (s, 3H), 2.41-2.39 (m, 2H), 2.20-2.12 (m, IH), 2.11 (s, 3H), 2.00-1.95 (m, lH), 1.89-1.84 (m, 3H), 1.75-1.45 (m, 14H), 1.42-0.92 (m, llH), 0.89 (s, 9H), 0.87-0.82 (m, IH), 0.62 (s, 3H), 0.00 (m, 6H). Step 13. Preparation of E14. To a solution ofE13 (230 mg, 0.48 mmol) in CH2Ch (6 mL) was added TF A ( 1 mL). The mixture was stirred at l 5°C for 30 min. TLC (petroleum ether: ethyl acetate =3: I) showed the starting material was consumed completely. The reaction was quenched with aqueous NaHCO3 (10 mL) and extracted with EtOAc (30 mL x 2). The organic layer was dried over anhydrous sodium sulfate. The organic phase was concentrated under vacuum. The crnde product was purified by colunm chromatography on silica gel ( eluent: petroleum ether: ethyl acetate= 10:1) to give E14 (166 mg, 95%) as white solid. 1H NMR (E14) : (400 MHz, CDC13) 4.12-4.1 l (m, lH), 3.52-3.42 (m, 2H), 3.30 (s, 3H), 2.58-2.53 (m, lH), 2.20-2.16 (m, 129 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 lH), 2.13 (s, 3H), 2.06-1.97 (m, 2H), 1.75-1.60 (m, SH), 1.59-1.I0 (m, IIH), 1.03-0.92 (m, lH), 0.90-0.82 (m, lH), 0.63 (s, 3H). Step 14. Preparation of EIS. To a solution ofE14 (2 g, 5.21 mmol) in MeOH (25 mL) was added HBr (5 drops) and Br2 (8 mL). The mixture was stirred at room temperature for 4 h. LCMS showed the starting material was consumed. The reaction was diluted with H2O (20 mL) and extracted with EtOAc (50 mL). The organic layer was dried over anhydrous sodium sulfate. The organic phase was concentrated under vacuum. The crude product was purified by colm1m chromatography on silica gel ( eluent: petroleum ether: ethyl acetate = 10: 1) give E 15 ( 1 g, 41 % ) as white solid. 1H-NMR showed there was 70% ofE15 and 30% ofE14. 1H NMR (EIS) :(400 MHz, CDCl3) 4.11-4.I0 (m, lH), 3.94-3.88 (m, 2H), 3.49-3.39 (m, 2H), 3.28 (s, 3H), 2.84-2.79 (m, IH), 2.20-2.15 (m, lH), 2.04-1.87 (m, 2H), 1.74-1.60 (m, SH), 1.57- 1.16 (m, 12H), 1.03-0.76 (m, 2H), 0.64 (s, 3H). Example 55. Preparation of Compounds 90 & 91. HO''. - H E15 90 O N-N , \\ Nv O N-N + ~N HO''' - H 91 To a solution of K2CO3 (952 mg, 6.9 mmol) in DMF (20 mL) was added 2H-l,2,3-triazole (969 mg, 13.8 mmol). The mixture was stirred at room temperature for 30 min, and then was added a solution of EIS (1 g, 2.3 mmol) in DMF (20 mL). The mixture was stirred at room temperature overnight. TLC (petroleum ether: ethyl acetate = 1: 1) showed the starting material was consumed completely. The reaction was diluted with EtOAc (50 mL) and washed with brine (50 mL). The organic layer was dried over anhydrous sodium sulfate. The organic phase was concentrated under vacuum. The crude product was purified by colW11TI chromatography on silica gel ( eluent: petroleum ether: ethyl acetate= 5:1) give 90 (204 mg, 21 %) as white powder and 91 (437 mg, 45%) as white powder. 1H NMR (90) : (400 MHz, CDC13) 7.70 (s, 2H), 5.31-5.20 (m, 2H), 4.13-4.12 (m, lH), 3.52-3.43 (m, 2H), 3.31 (s, 3H), 2.63-2.58 (m, IH), 2.24-2.22 (m, lH), 2.12- 130 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 1.97 (m, 2H), 1.78-1.62 (m, 9H), 1.54-1.10 (m, l0H), 1.09-0.96 (m, lH), 0.91-0.84 (m, lH), 0.75 (s, 3H). 1H NMR (91) : (400 MHz, CDC13) 7.75 (s, lH), 7.63 (s, IH), 5.27-5.11 (m, 2H), 4.13-4.10 (m, lH), 3.50-3.40 (m, 2H), 3.28 (s, 3H), 2.67-2.64 (m, lH), 2.26-2.18 (m, IH), 2.08- 1.92 (m, 2H), 1.74-1.25 (m, 15H), 1.19-0.87 (m, 8H), 0.69 (s, 3H). Example 56. Preparation of Compound 92. Br ( 0 H~o 0 O N-N ' ~ 0 Ho" -H D15 92 A mixture of E15 (200 mg, 0.45 mmol), K2CO3 (188 mg, 1.36 mmol), ethyl lH-pyrazole-3- carboxylate (317 mg, 2.27 mmol) and DMF (2 mL) were was stirred at 25 °C for 12 hours. TLC showed the reaction was finished. The mixture was diluted with EtOAc (20 mL), washed with brine (30mL *3), and the organic layer was dried over anhydrous Na2SO4, and then concentrated to give crude product. It was purified by column chromatography (petroleum ether: ethyl acetate = 4:1) to give the 92 (96 mg, 44.4 %) as a white solid. 1H NMR (92): (400 MHz, CDC13) 8 7.42 (d, J=2.4 Hz, lH), 6.86 (d, J=2.4 Hz, lH), 5.05-4.95 (m, 2H), 4.39 (q, J=7.2Hz, 2H), 4.20-4.10 (m, lH), 3.49-3.39 (m, 2H), 3.28 (s, 3H) ,2.60-2.55 (m, IH), 2.22 -1.91 (m, 3H), 1.75-1.50 (m, 6H), 1.45-1.10 (m, 15H), 1.05-0.92 (m, lH), 0.90-0.84 (m, lH), 0.70 (s, 3H) Example 57. Preparation of Compound 93. Br E15 Date Rec;ue / Date Received 2024-04-12 0 CH3CN,60 °C, 10 h 46.8% 131 93 WO 2015 / 027227 PCT / US2014 / 052417 To a solution ofE15 (200 mg, 0.47 mmol) in CR,CN (15 mL) at 29 °C, then K2CO3 (194.01 g, 1.4 mmol) and 4-methyl-lH-pyrazole (192.09 mg, 2.34 mmol) was added in the mixture at 29 0C. The solution was stirred at 60 °C for 10 h. After the TLC showed that the starting material was consumed completely, then the mixture was concentrated. The mixture was extracted with CH2Ch (30 mL) and NaCl (aq) (20 mL *2). The combined organic layers were dried over Na2SO4 and concentrated to give crude product. The product was purified by column chromatograph on silica gel eluted with (petroleum ether / ethyl acetate=IO: 1 to 6: 1) to give 93 (94.4 mg, yield: 46.81 %). 1H NMR (93): (400 MHz, CDC13) 8 7.36 (s, lH ), 7.18 (s, IH), 4.85 (dd, J= 34 Hz, J= 17.6 Hz, 2H), 4.15-4.10 (m, lH), 3.47 (dd, J= 28.8Hz, J= 10 Hz, 2H), 3.31 (s, 3H), 2.62-2.55 (m, lH), 2.28 - 2.18 (m, IH), 2.13-1.97 (111, SH), 1.78-1.64 (m, 8H), 1.48-1.13 (m, IOH), 1.04-0.84 (m, 2H), 0.71 (s, 3H) Example 58. Preparation of Compounds 94, 95 & 96. 0 Br ~,,N;>-- HNJ - ,. Cs2C03, DMF HO' - H E15 94 95 96 To a solution of E15 (150 mg, 0.35 mmol) in DMF (6 mL) was added Cs2CO3 (343 mg, 1.05 mmol) and 4-methyl-l, 2, 3-triazole (145 mg, l.75 mmol) at 28 °C. The reaction mixture was stirred at the same temperature for 6 h. TLC showed that the starting material was consumed completely. The mixture was poured into water (20 mL), and extracted with EtOAc (10 mL *2). The combined organic layers were dried over Na2SO4 and concentrated to give crude product. The crude product was purified by pre-HPLC to give pure 94 (35.8 mg) and a mixture of95 and 96 (20 mg). Then the mixtue was purified by SFC to give 95 (3.9 mg) and 96 (5.6 mg). Total yield: 23. 7%. The structure of the three targets was confirmed by NOE. 1H NMR (94): (400 MHz, CDC13) 8 7.42 (s, IH), 5.19-5.08 (m, 2H), 4.13-4.09 (m, IH), 3.53- 3.38 (m, 2H), 3.29 (s, 3H), 2.61-2.54 (m, lH), 2.33 (s, 3H), 2.26-2.15 (m, lH), 2.11-1.95 (m, 2H), 1.78-1.61 (m, 9H), 1.52-1.06 (m, 9H), 1.04-0.81 (m, 2H), 0.72 (s, 3H). 132 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 1H NMR (95): (400 MHz, CDC13) 8 7.50 (s, lH), 5.23-5.05 (m, 2H), 4.15-4.06 (m, lH), 3.58- 3.42 (m, 2H), 3.31 (s, 3H), 2.71-2.61 (m, lH), 2.33-2.14 (m, 6H), 2.12-1.96 (m, 4H), 1.34-1.18 (m, IOH), 1.06-0.83 (m, 6H), 0.70 (s, 3H) . 1H NMR (96): (400 MHz, CDC13) 8 7.35 (s, lH), 5.22-5.02 (m, 2H), 4.15-4.10 (m, lH), 3.56- 3.39 (m, 2H), 3.31 (s, 3H), 2.67-2.61 (m, lH), 2.39 (s, 3H), 2.29-1.88 (m, l0H), 1.33-1.22 (m, I0H), 1.02-0.97 (m, IH), 0.92-0.84 (m, 2H), 0.70 (s, 3H). Example 59. Preparation of 97 and 98. Br A11 N,(¾,-F N~ ~ ,:;; F BB•1 0 97 N-N '~ N" I ,,___ F F HCf H 98 To a solution of All (360 mg, 0.82 mmol) in acetone (2.5 mL) was added 5,6-difluoro-2Hbenzo[ d] [l,2,3]triazole (BB-1) (190 mg, 1.23 mmol) and K2CO3 (230 mg, 1.64 mmol). The mixture was stirred at 30 °C for 3 hours. TLC showed the reaction was compeleted. To the mixture was added water (2 mL), extracted with EtOAc (5 mL * 2). The combined organic layer was concentrated in vacuum, purified by prep-HPLC to give 97 (25 mg, 6%) and 98 (141 mg, 33%) as a white solid. 1H NMR (97): (400 MHz, CDCb) 8 7.59 (t, J = 8.4 Hz, 2H), 5.53-5.42 (m, 2H), 3.48-3.36 (m, 2H), 3.29 (s, 3H), 2.65 (t, J = 8.4 Hz, IH), 2.27-2.02 (m, 3H), 1.80-0.85 (m, 23H), 0.75 (s, 3H). LC:MS (97): tR = 1.366 min in 2 min chromatography, 10-80AB, purity 96.3%, MS ESI calcd. for C29H40F2N3O3 [M+Ht 516, found 498([M+H-18]\ 1H NMR (98): (400 MHz, CDCh) 8 7.83 (t, .J= 8.0 Hz, lH), 7.12 (t, .J = 7.6 Hz, lH), 5.43-5.32 (m, 2H), 3.50-3.37 (m, 2H), 3.30 (s, 3H), 2.80-2.68 (m, lH), 2.30-2.02 (m, 3H), 1.80-0.85 (m, 23H), 0. 75 (s, 3H). LCMS (98): tR = 1.317 min in 2 min chromatography, I 0-80AB, purity 99.6%, MS ESI calcd. for C29H40F2N3Q3 [M+Ht 516, found 516. 133 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Scheme A. General Procedure for the Preparation of Compounds 99-146. A11 .-..._ Br 0 Ho' H Example Structure of BB-n Product(s) NMR / LC-MS 0 Hd H 99 60 BB-16 0 A 100 Date Rec;ue / Date Received 2024-04-12 N-N ,~~ "'Y Cl 1H NMR(400 MHz, CDCl3) o 8.74 (d, .J = 2.4 Hz, lH), 8.23 (d, .J = 2.4 Hz, IH), 5.60-5.49 (m, 2H), 3.49-3.37 (m, 2H), 3.30 (s, 3H), 2.68 (t, .J = 8 Hz, lH), 2.23- 2.03 (m, 3H), 1.79-0.88 (m, 23H), 0.77(s, 3H). LCMS tR = 1.486 min in 2 min chromatography, 10-80AB, purity 100.0%, MS ESI calcd. for C28H40ClN4O3 [M+Ht 515, found 515. 1H NMR (400 MHz, CDCh) & 8.59 (d, .J = 2.0 Hz, lH), 8.37 (d, .J = 2.0 Hz, lH), 5.56-5.45 (m, 2H), 3.51-348 (m, IH), N !'J-<,°'\ Cl 3.40-3.37 (m, lH), 3.31 (s, 3H), 2.27 (t,J N, •.. ~ N 134 = 8.8 Hz, IH), 2.25-2.22 (m, 2H), 2.10- 1.95 (m, IH), 1.78-1.28 (m, 12H), 1.28- 0.80 (m, l lH), 0.79-0.72 (s, 3H). LCMS tR = 1.495 min in 2 min chromatography, 10-S0AB, purity 99.7%, WO 2015 / 027227 0 Cl f-J-v~ N,,N N Hd A 101 0 N 61 HN;tN) N:(j BB-19 Hd A 102 135 Date Rec;ue / Date Received 2024-04-12 PCT / US2014 / 052417 MS ESI calcd. for C28H40ClN4O3 [M+Ht 515, found 515. 1H NMR (400 MHz, CDCh) o 8.68 (s, lH), 7.77 (s, lH), 5.50-5.36 (m, 2H), 3.49-3.47 (m, lH), 3.39-3.37 (m, lH), 3.30 (s, 3H), 2.60-2.80 (111, lH), 2.22-2.03 (m, 3H), 1.90-1.38 (m, 13H), 1.34-1.18 (m, 8H), 1.15-0.73 (m, SH). LCMS tR = 1.446 min in 2 min chromatography, 10-80AB, purity 98.9%, MS ESI calcd. for C28H40ClN4O3 [M+Ht 515, found 515. 1H NMR ( 400 MHz, CDCb) o 8.59 ( d, J = 2.4 Hz, lH), 8.46 (d,J= 2.4 Hz, IH), 8.35 (s, lH), 5.38-5.24 (m, 2H), 3.50-3.48 (111, lH), 3.40-3.37 (m, lH), 3.31 (s, 3H), 2. 74-2. 71 (m, lH), 2.25-2.22 (m, 3H), 1.80-0.88 (111, 23H), 0.78-0.65 (m, 3H) LCMS tR = 1.376 min in 2 min chromatography, 10-80AB, purity 92.9%, MS ESI calcd. for C28H41N4O3 [M+Hf 481, found 481. 62 WO 2015 / 027227 HN!~'J t)-~ ~' OCF3 BB-4 Ho' R Date Rec;ue / Date Received 2024-04-12 - 0 PCT / US2014 / 052417 1H NMR (400 MHz, CDCb) 8 8.64 (d, J = 1.6 Hz, lH), 8.56 (d,J= 1.6 Hz, lH), 8.26 (s, lH), 5.37-5.21(m, 2H), 3.49-3.46 (m, lH), 3.39-3.37 (m, lH), 3.33 (s, 3H), N, 1 2.75-2.65 (m, lH), 2.17-2.01 (m, 3H), N~ ~ N NJ 1.76-0.80 (m, 23H), 0.74 (s, 3H) 103 LCMS tR = 1.281 min in 2 min chromatography, 10-80AB, purity 99. I%, MS ESI calcd. for C28H41N4O3 [M+Hf 481, found 481. 0 N-N 1H NMR (400 MHz, CDCh) 8 7.90 (d, J = 2.0Hz, lH), 7.72 (s, lH), 7.28-7.26 (m, lH), 5.58-5.48 (m, 2H), 3.49-3.47 (m, IH), 3.49-3.47 (m, lH), 3.29 (s, 3H), 2.67 NC F (t, J= 8.4 Hz, lH), 2.25-1.80 (m, 3H), ~ I o)<_; 1. 75-0.81 (m, 23H), 0. 77 (s, 3H). 104 F 0 0-f- ~Ji' FF N,-. .. ~ N 144 136 LCMS tR = 1.618 min in 2 min chromatography, 10-80AB, purity 98.7%, MS ESI calcd. for C30H41F3N3O4 [M+Ht 564, found 564. 1H NMR(400 MHz, CDC13) 8 8.10 (d,J = 9.2 Hz, lH), 7.27-7.25 (m, lH), 7.19 (s, IH), 5.54-5.35 (m, 2H), 3.50-3.48 (m, lH), 3.40-3.37 (m, lH), 3.30 (s, 3H), 2.75-2.73 (m, lH), 2.27-2.01 (m, 3H), 1.80-0.80 (m, 23H), 0.74 (s, 3H). LCMS tR = 1.002 min in 1.5 min chromatography, 5-95AB, purity 96.4%, WO 2015 / 027227 0 H N(::--. \Jc 63 :::,..__ I Cl BB-21 Hcf A Date Rec;ue / Date Received 2024-04-12 PCT / US2014 / 052417 MS ESI calcd. for C30H41F3N3O4 [M+Ht564, found 564. 1H NMR (400 MHz, CDC13) 8 7.94 (s, IH), 7.40-7.34 (m, 2H), 5.49-5.38 (m, 2H), 3.50-3.48 (m, lH), 3.40-3.37 (m, ~f lH), 3.31 (s, 3H), 2.76-2.73(m, IH), 2.27- N~_. >-=~, ro / -F 2.01 (m, 3H), 1.80-0.80 (m, 23H), 0.75 (s, N 3H). 146 105 137 LCMS tR = 0.998 min in 1.5 min chromatography, 5-95AB, purity 94.7%, MS ESI calcd. for C30H41F3N3O4 [M+H]+564, found 564. 1H NMR (400 MHz, CDCh) 8 7.98 (s, lH), 7.72 (d, J= 1.6 Hz, IH), 7.34- 7.3l(m, lH), 7.14 (d, J = 8.8 Hz, lH), 5.17-5.07 (m, 2H), 3.50-3.47 (m, lH), 3.39-3.37 (111, lH), 3.30 (s, 3H), 2.64 (t, J = 8.4 Hz, IH), 2.12-2.01 (m, 3H), 1.74- 0.80 (m, 23H), 0.74 (s, 3H). LCMS tR = 1.54 7 min in 2 min chromatography, 10-80AB, purity 100.0%, MS ESI calcd. for C30H42ClN2O3 [M+Ht 513, found 513. WO 2015 / 027227 PCT / US2014 / 052417 1H NMR (400 MHz, CDCb) 8 7.88 (s, IH), 7.64-7.62 (m, 2H), 7.23-7.20 (m, lH), 5.24-5.12 (m, 2H), 3.49-3.46 (m, 0 IH), 3.39-3.47 (m, lH), 3.29 (s, 3H), ~D 2.70-2.60 (m, lH), 2.18-0.79 (m, 26H), N:c-- ~ \ Cl 0.73 (s, 3H). 106 LCMS tR = 1.506 min in 2 min chromatography, 10-80AB, purity 99.8%, MS ESI calcd. for C30H42CIN2O3 [M+Ht 513, found 513. 1H NMR (400 MHz, CDCh) 8 8.00 (s, lH), 7.38-7.35 (m, lH), 7.17-7.14 (m, 2H), 5.18-5.09 (m, 2H), 3.50-3.47 (m, 0 IH), 3.39-3.37 (m, lH), 3.30 (s, 3H), 2.64 N:0-F (t, J = 8.4 Hz, lH), 2.21-2.03 (m, 3H), 1.75-0.80 (m, 23H), 0.74 (s, 3H). H◊- A 107 LCMS tR = 1.479 min in 2 min chromatography, 10-80AB, purity 99.5%, HN ~ MS ESI calcd. for C30H42FN2O3 64 F [M+Ht 497, found 497. BB-23 1H NMR (400 MHz, CDC13) 8 7.89 (s, 0 IH), 7.68(dd, J = 4.4 Hz, J = 8.6 Hz, lH), !"°' 7.23-7.20(m, IH), 7.10-7.08 (m, lH), N:,._ I 5.25-5.15 (m, 2H), 3.49-3.46 (m, lH), ~ F 3.39-3.37 (m, lH), 3.30 (s, 3H), 2.65 (t, J Hcf A 108 = 8.4 Hz, lH), 2.30-2.08 (m, 3H), 1.75- 0.80 (m, 23H), 0. 74 (s, 3H). LCMS tR = 1 .455 min in 2 min 138 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 0 109 65 HN ,N..._,~ ,N...,, ,,,::::, Cl F BB-8 0 138 Date Rec;ue / Date Received 2024-04-12 N-N -~ N" \ ::,.,_ F Cl PCT / US2014 / 052417 chromatography, I 0-S0AB, purity 96.5%, MS ESI calcd. for C30H42FN2O3 [M+Ht 497, found 497. 1HNMR (400 MHz, CDCb) o 7.63 (d, J= 8.6 Hz, lH), 7.36 (dd, J = 6.4 Hz, J = 8.6 Hz, lH), 5.54-5.52 (m, 2H), 3.49-3.47 (m, IH), 3.40-3.37 (m, lH), 3.30 (s, 3H), 2.70-2.60 (m, IH), 2.30-1.90 (m, 3H), 1.80-0.80 (m, 23H), 0.77 (s, 3H). LCMS tR = 1.434 min in 2 min chromatography, I 0-S0AB, purity 99.2%, MS EST calcd. for C29H40ClFN3O3[M+Ht532, found 5 l 4[M+H-H2Or. 1H NMR (400 MHz, CDCh) o 7.48 (dd, J= 6.0 Hz, J=8.8 Hz, lH), 7.07 (d, J= 7.6 Hz, IH), 5.46-5.36 (m, 2H), 3.50-3.48 (m, lH), ---.c: 3.40-3.37 (m, lH), 3.30 (s, 3H), 2.72 (t, J = N~ ~ I? Cl 'N 8.6 Hz, lH), 2.30-0.80 (m, 26H), 0.74 (s, 3H). F 139 LCMS tR = 1.374 min in 2 min chromatography, 10-80AB, purity 99.8%, MS ESI calcd. for C29H40CIFN3O3 [M+Ht 532, found 532. WO 2015 / 027227 PCT / US2014 / 052417 1H NMR (400 MHz, CDCb) 8 8.01 (s, IH), 7.68 (dd, J = 5.2 Hz, J = 8.4 Hz, lH), 6.96-6.84 (m, 2H), 5.14-5.04 (m, 2H), 0 3.66-3.47 (m, IH), 3.39-3.37 (m, lH), F f'J:O: 3.29 (s, 3H), 2.65 (t, J= 8.4 Hz, lH), N:--. 2.09-2.03 (m, 3H), 1.80-0.80 (m, 23H), 0.74 (s, 3H). Hcf H 110 LCMS tR = 1.480 min in 2 min chromatography, 10-80AB, purity 100.0%, MS ESI calcd. for ~ F C30H42FN2O3 [M+Ht 497, found 497. HN 66 _✓,:; BB-22 1H NMR (400 MHz, CDCh) 8 7.93 (s, lH), 7.64 (dd, J = 5.4 Hz, J = 8.8 Hz, lH), 7.30-7.20 (m, lH), 6.92-6.87 (m, IH), 0 5.23-5.11 (m, 2H), 3.49-3.46 (m, lH), f'J~ 3.39-3.37 (m, IH), 3.29 (s, 3H), 2.65 (t, J N,._ \ = 8.4 Hz, lH), 2.08-1.90 (m, 3H), 1.80- :::,... 0.80 (m, 23H), 0.73 (s, 3H). H6 H F 111 LCMS tR = 1.446 min in 2 min chromatography, 10-80AB, purity 96.7%, MS ESI calcd. for C30H42FN2O3 [M+Ht 497, found 497. 1H NMR (400 MHz, CDCh) 8 8.01 (s, 0 Cl ~~ '°"""Cl f'J:e;-: lH), 7.66(d,.J=8.4Hz, 1H),7.20(s, HN IH), 7.13 (d,J= 8.6, lH), 5.15-5.05 (m, 79 --- ~ N" BB-20 2H), 3.50-3.47 (m, lH), 3.40-3.37 (m, Hd 112 lH), 3.30 (s, 3H), 2.66 (t, J = 8.4, IH), 2.20-2.05 (m, 4H), 1.80-0.80 (m, 22H), 140 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 113 68 Date Rec;ue / Date Received 2024-04-12 PCT / US2014 / 052417 0.74 (s, 3H). LCMS tR = 1.548 min in 2 min chromatography, 10-80AB, purity 100.0%, MS ESI calcd. for C30H42ClN2O3 [M+Hf 513, found 513. 1H NMR (400 MHz, CDCb) 8 7.93 (s, IH), 7.70 (s, lH), 7.61 (d, .I= 8.6 Hz, lH), 7.07-7.04 (m, lH), 5.30-5.20 (m, 2H), 3.49-3.46 (m, lH), 3.39-3.37 (m, lH), 3.29 (s, 3H), 2.65 (t, .I= 8.4 Hz, lH), 2.10-0.80 (m, 26H), 0. 72 (s, 3H). LCMS tR = 1.517 min in 2 min chromatography, 10-80AB, purity 99.7%, MS ESI calcd. for C30H42ClN2O3 [M+Hf 513, found 513. 1HNMR (400 MHz, CDC13) 8 7.39-7.36 (m, lH), 7.06-7.03 (m, lH), 5.54-5.35 (m, 2H), 3.50-3.48 (m, lH), 3.40-3.37 (m, lH), 3.30 (s, 3H), 2.74-2.70 (m, lH), * ~ F N" 2.27-2.01 (m, 3H), 1.80-0.80 (m, 23H), N F 141 0.74 (s, 3H). LCMS tR = 0.967 min in 1.5 min chromatography, 5-95AB, purity 100.0%, MS ESI calcd. for C29H40F2N3O3 [M+Hf 516, found 516. WO 2015 / 027227 HW~, 69 I N N\ Date Rec;ue / Date Received 2024-04-12 0 H 116 142 PCT / US2014 / 052417 1HNMR(400 MHz, CDCl,) o 7.82-7.79 (m, lH), 7.24-7.21 (m, lH), 5.54-5.35 (m, 2H), 3.50-3.48 (m, lH), 3.40-3.37 (m, IH), 3.29 (s, 3H), 2.75-2.73 (m, IH), 2.27-2.01 (m, 3H), 1.80-0.80 (m, 23H), 0.75 (s, 3H). LCMS tR = 0.986 min in 1.5 min chromatography, 5-95AB, purity 96.8%, MS ESI calcd. for C29H40F2N3O3 [M+Ht 516, found 516. 1HNMR (400 MHz, CDC13) 8 7.65-7.62 (m, IH), 7.29-7.27 (m, IH), 5.54-5.35 (m, 2H), 3.50-3.48 (m, lH), 3.40-3.37 (m, IH), 3.29 (s, 3H), 2.74-2.70 (m, lH), 2.27-2.01 (m, 3H), 1.80-0.80 (m, 23H), 0.76 (s, 3H). LCMS tR = 1.012 min in 1.5 min chromatography, 5-95AB, purity 95.6%, MS ESI calcd. for C29H40F2N3O3 [M+Ht 516, found 498[M+H-H2Ot. H NMR (400 MHz, CDCb) o 5.39-5.29 (m, 2H), 3.47-3.45 (m, IH), 3.38-3.35 (m, lH), 3.28 (s, 3H), 2.74-2.60 (m, IH), 2.55 (s, 3H), 2.27-2.01 (m, 3H), 1.80-0.80 (m, 23H), 0.72 (s, 3H). LCMS tR = 0.909 min in 1.5 min chromatography, 5-95AB, purity 100.0%, MS ESI calcd. for C2sH41N4O3 [M+Ht WO 2015 / 027227 PCT / US2014 / 052417 445, found 445. H NMR (400 MHz, CDCh) 8 5.14-5.04 (m, 2H), 3.48-3.45 (m, lH), 3.38-3.35 (m, 0 lH), 3.28 (s, 3H), 2.74-2.60 (m, lH), 2.46 N-( (s, 3H), 2.27-2.01 (m, 3H), 1.80-0.80 (m, I N N",, 23H), 0.69 (s, 3H). N LCMS tR = 0.882 min in 1.5 min 117 chromatography, 5-95AB, purity 94.3%, MS ESI calcd. for C2sauN4O3 [M+Ht 445, found 427[M+H-18t. H NMR (400 MHz, CDCh) 8 7.47 (d, J = 2.4 Hz, 1 H), 6. 72 ( d, J = 2.0 Hz, IH), 5.04-4.89 (m, 2H), 3.47-3.45 (m, lH), 0 3.38-3.35 (m, lH), 3.28 (s, 3H), 2.59 (t, J H ~r = 8.4 Hz, IH), 2.27-2.01 (m, 3H), 1.80- N-N N:,, 70 )L) 0.80 (m, 23H), 0.68 (s, 3H). NC II N 118 LCMS tR = 1.34 7 min in 2 min chromatography, 10-80AB, purity 100.0%, MS ESI calcd. for C21H40N3O3 [M+Ht 454, found 436[M+H-18t. H 0 Cl H NMR (400 MHz, CDCb) o 7.99 (dd, J ,N-N f-lj)-: = 2.4 Hz, J= 6.4 Hz, IH), 7.34-7.32 (m, 71 Nnl N,:- N 2H), 5.42-5.31 (m, 2H), 3.49-3.47 (m, ~' IH), 3.39-3.36 (m, lH), 3.30 (s, 3H), Cl 119 2.73-2.60 (m, lH), 2.30-1.90 (m, 3H), 143 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 120 0 121 Date Rec;ue / Date Received 2024-04-12 PCT / US2014 / 052417 1.80-0.80 (m, 23H), 0.74 (s, 3H). LCMS tR = 1.430 min in 2 min chromatography, 10-80AB, purity 99.5%, MS ESI calcd. for C29H41ClN3O3 [M+Ht 514, found 514. H NMR (400 MHz, CDCh) 8 7.87 (s, lH), 7.82 (d, J = 8.6 Hz, 2H), 7.36-7.33 (m, IH), 5.56-5.45 (m, 2H), 3.49-3.47 (m, IH), 3.40-3.37 (m, lH), 3.30 (s, 3H), 2.73-2.60 (m, IH), 2.30-1.90 (m, 3H), 1.80-0.80 (m, 23H), 0. 76 (s, 3H). LC.MS tR = 1.473 min in 2 min chromatography, 10-80AB, purity 98.2%, MS ESI calcd. for C29fii1ClN3O3 [M+H]" 514, found 496[M+H-18t. H NMR (400 MHz, CDCh) 8 8.05 (s, IH), 7.44 (dd, J= 1.6 Hz, J= 8.8 Hz, lH), 7.26 (d, J = 8.4 Hz, IH), 5.49-5.38 (m, 2H), 3.49-3.46 (m, IH), 3.38-3.36 (m, -✓-=\ lH), 3.29 (s, 3H), 2.80-2.60(m, lH), 2.27- rt Vc1 2.01 (m, 3H), 1.80-0.80 (m, 23H), 0.72 (s, N 144 3H). LC.MS tR = 1.409 min in 2 min chromatography, 10-80AB, purity 100.0%, MS ESI calcd. for C29H41ClN3O3 [M+Ht 514, found 514. WO 2015 / 027227 122 H 72 Fi\ F 123 124 Date Rec;ue / Date Received 2024-04-12 145 PCT / US2014 / 052417 H NMR (400 MHz, CDCb) 8 6.89-6.86 (m, lH), 6.79 (dd, J= 1.6 Hz, J= 7.2 Hz, lH), 5.52-5.32 (m, 2H), 3.49-3.47 (m, lH), 3.39-3.36 (m, IH), 3.29 (s, 3H), 2.71 (t,.J= 8.8 Hz, lH), 2.27-2.01 (m, 3H), 1.80-0.80 (m, 23H), 0.74 (s, 3H). LCMS tR = 0.982 min in 1.5 min chromatography, 5-95AB, purity 100.0%, MS ESI calcd. for C29fLioF2N3O3 [M+Ht 516, found 516. H NMR (400 MHz, CDCb) 8 7.31 (dd, J = 1.6 Hz, J = 8.4 Hz, lH), 6.94-6.88 (m, lH), 5.57-5.46 (m, 2H), 3.49-3.47 (m, lH), 3.40-3.37 (m, lH), 3.30 (s, 3H), 2.74-2.70 (m, lH), 2.27-2.01 (m, 3H), 1.80-0.80 (111, 23H), 0.76 (s, 3H). LCMS tR = 1.015 min in 1.5 min chromatography, 5-95AB, purity 99 .0%, MS ESI calcd. for C29H40F2N3O3 [M+Ht 516, found 498[M+H-I8t. H NMR (400 MHz, CDCh) 6 7.52 (dd, J = 1.6 Hz, J= 8.0 Hz, lH), 7.01-6.95 (m, IH), 5.54-5.35 (m, 2H), 3.49-3.47 (m, IH), 3.38-3.36 (m, lH), 3.29 (s, 3H), 2.74-2.70 (m, lH), 2.27-2.01 (m, 3H), 1.80-0.80 (m, 23H), 0.72 (s, 3H). LCMS tR = 0.985 min in 1.5 min WO 2015 / 027227 0 73 125 74 126 Date Rec;ue / Date Received 2024-04-12 ~x N~ F F F 146 PCT / US2014 / 052417 chromatography, 5-95AB, purity 95.7%, MS ESI calcd. for C29H40F2N3O3 [M+Ht 516, found 498[M+H-18t. H NMR (400 MHz, CDCl3) o 7.45 (d, .J = 1.2 Hz, lH), 6.58 ( d, J = 2.0 Hz, lH), 5.01-4.91 (m, 2H), 3.47-3.45 (m, lH), 3.38-3.35 (m, lH), 3.28 (s, 3H), 2.58 (t, J = 8.8 Hz, lH), 2.27-2.01 (m, 3H), 1.80- 0.80 (m, 23H), 0.69 (s, 3H). LCMS tR = 1.389 min in 2 min chromatography, I 0-80AB, purity 99 .3 %, MS ESI calcd. for C27H40F3N2O3 [M+Ht 497, found 479[M+H-I8t. H NMR(400 MHz, CDCb) o 7.48 (t,J= 8.0 Hz, lH), 7.27-7.22 (m, 2H), 5.50-5.40 (m, 2H), 3.50-3.49 (m, lH), 3.39-3.37 (m, IH), 3.30 (s, 3H), 2.75-2.65 (m, lH), 2.27-2.01 (111, 3H), 1.80-0.80 (m, 23H), 0.75 (s, 3H). LCMS tR = 1.459 min in 2 min chromatography, l 0-80AB, purity 100.0%, MS ESI cakd. for C10H41F3N3O4 [M+Ht 564, found 564. WO 2015 / 027227 PCT / US2014 / 052417 1HNMR (400 MHz, CDC13) 8 7.85 (s, lH), 7.81 (s, lH), 5.04-4.87 (m, 2H), 3.49-3.45 (m, IH), 3.38-3.36 (m, 1H), H ~ O N-N 3.28 (s, 3H), 2.60 (t, J = 8.8 Hz, lH), N-N 2.17-2.01 (m, 3H), 1.80-0.80 (m, 23H), 75 l{ y 0.68 (s, 3H). CN CN HO H BB-27 127 LCMS tR = 1.304 min in 1.5 min chromatography, 10-80AB, purity 98.5%, MS ESI calcd. for C27H40N3O3 [M+Ht 454, found 436[M+H-l8f. Example 76. Preparation of Compounds 154 and 155. 0 0 Y' N N:,.. N Br HN:() NJ 0 N N H 154 I N:() 0 K2CO3, acetone, 25 °c H D6 H 155 To a solution of compound D6 (300 mg, 0.679 mmol) in acetone (5 mL) was added K2CO3 (186 mg, 1.35 rnmol) and 2H-pyrazolo[3,4-b]pyrazine (120 mg, 1.01 mrnol). After stirring at 25 oC for 3 h, LCMS showed the reaction was complete, one product (27%) and one product (21 %). The reaction mixture was concentrated in vacuum to remove most of the solvent to give the residue. The residue was dissolved in EtOAc (50 mL), and washed with water (50 mL). The reaction mixture was extracted with EtOAc (50 mL * 3). The organic layers were combined and concentrated in vacuum to give the cmde product. The cmde product was purified by prep. HPLC (FA) to give compound 154 (4 mg, 1.22%) as a whte solid and compound 155 (4 mg,1.22%) as white solid. 147 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 1H NMR (154) (yield 1.2%): (400 MHz, CDCh) o 8.64 (d, J = 2.0 Hz, lH), 8.56 (d, J = 2.0 Hz, 1 H), 8.26 (s, lH), 5.40-5.19 (m, 2H), 3.54 (d, J = 8.4 Hz, lH), 3.33 (s, 3H), 3.20 (d, J = 9.2 Hz, IH), 2.68 (t, J= 8.8 Hz, IH), 2.30-1.20 (m, 26H), 0.72 (d, J= 13.2 Hz, 3H). LCMS tR = 1.155 min in 2 min chromatography, I 0-80AB, purity 96.2%, MS ESI calcd. for C28H41N4O3 [M+Hf 481, found 481. 1H NMR (155) (yield 1.2%): (400 MHz, CDCh) o 8.59 (d, J= 2.4 Hz, IH), 8.44 (d,J= 2.0 Hz, lH), 8.34 (s, lH), 5.36-5.24 (m, 2H), 3.55 (d, J = 9.2 Hz, lH), 3.34 (s, 3H), 3.22 (d, J = 9.2 Hz, IH), 2.68 (t, J= 8.8 Hz, IH), 2.30-1.20 (m, 26H), 0.71 (s, 3H). LCMS tR = 0.854 min in 1.5 min chromatography, 10-80AB, purity 96.8%, MS ESI calcd. for C2sH41N4O3 [M+Hf 481, found 463[M+H-18r. Example 77. Preparation of Compound 156. Br Hq y CN CN K2C03 H H D6 156 1H NMR (156) (yield 41 %): (400 MHz, CDCb) o 7.88 (s, IH), 7.83 (s, lH), 5.07-4.87 (m, 2H), 3.55 (d, J= 9.0 Hz, lH), 3.35 (s, 3H), 3.22 (d, J= 9.0 Hz, lH), 2.65-2.58 (m, IH), 2.28-2.15 (m, lH), 2.10-2.01 (m, IH), 1.97-1.91 (m, 2H), 1.82-1.40 (m, 14H), 1.35-1.09 (m, 8H), 0.68 (s, 3H). LCMS tR = 2.744 min in 4 min chromatography, 10-80AB, purity 100.0 %, MS ESI calcd. for C27H39N3O3Na [M+Nar 477, found 477. Example 78. Preparation of Compound 147. 148 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 Br D6 HN~ 'N~CI PCT / US2014 / 052417 1H NMR (147) (yield 4 %): (400 MHz, CDCh) 8 8.03 (s, lH), 7.68 (d, J= 8.5 Hz, lH), 7.22 (s, IH), 7.15 (d, J = 8.5 Hz, IH), 5.20-5.03 (m, 2H), 3.57 (d, J = 9.0 Hz, 1H), 3.36 (s, 3H), 3.23 (d, J = 9.0 Hz, lH), 2.65 (t,J= 8.7 Hz, IH), 2.30-2.09 (m, 2H), 1.99-1.91(m, 2H), 1.77-1.45 (m, 14H), 1.34-1.14 (m, SH), 0.73 (s, 3H). LCMS tR = 3.185 min in 4 min chromatography, 10- S0AB, purity 100.0 %, MS ESI calcd. for C3(JH42ClN2O::i [M+Hf 513, found 513. Example 79. Preparation of Compound 157. 0 Br D6 157 1H NMR (157) (yield 3.5%): (400 MHz, CDCh) o 7.47-7.43 (m, lH), 7.14-7.06 (m, 2H), 5.49- 5.38 (m, 2H), 3.55 (t, .l = 9.2 Hz, IH), 3.36 (d, .l = 4.0 Hz, 3H), 3.25 (d, .l = 8.8 Hz, lH), 2.73 (s, IH), 2.30-1.20 (m, 26H), 0.74 (s, 3H). LCMS tR = 0.931 min in 1.5 min chromatography, 5- 95AB, purity 96.8%, MS ESI calcd. for C29H41FN3O3 [M+Hf 498, found 520[M+Naf. Example 80. Preparation of Compound 158. 149 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 Br N----N V--cN 06 158 1H NMR (158) (yield 18 %): (400 MHz, CDCb) o 7.50 (d, J= 2.5 Hz, lH), 6.75 (d, J= 2.5 Hz, lH), 5.08-4.90 (m, 2H), 3.55 (d, J = 9.0 Hz, IH), 3.37-3.31 (m, 3H), 3.22 (d, J = 9.0 Hz, IH), 2.64-2.57 (m, IH), 2.28-01 (m, 2H), 1.99-1.91 (m, 2H), 1.84-1.40 (m, 14H), 1.39 - 1.08 (m, 8H), 0.67 (s, 3H). LCMS tR = 2.821 min in 4 min chromatography, 10-80AB, purity 100.0 %, MS ESI calcd. for C21H39N3O3Na [M+Nat 476, fom1d 476. Example 81. Preparation of Compound 159. Br N--N F ~F F 06 159 1H NMR (159) (yield: 13 %): (400 MHz, CDCb) 8 7.48 (s, lH), 6.60 (d, J = 2.0 Hz, 1H), 5.07- 4.88 (m, 2H), 3.56 (d, J = 9.0 Hz, lH), 3.35 (s, 3H), 3.22 (d, J = 9.0 Hz, lH), 2.65-2.54 (m, IH), 2.28-2.15 (m, lH), 2.10-2.02 (m, lH), 1.99-1.90 (m, 2H), 1.84-1.37 (m, 14 H), 1.36-1.07 (m, 8 H), 0.68 (s, 3H). LCMS tR = 3.049 min in 4 min chromatography, 10-80AB, purity 100.0 %, MS ESI calcd. for C21H40F3N2O3 [M+H]" 497, found 479 [M+H-H2O]1. Example 82. Preparation of Compound 160. 150 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 Br 06 PCT / US2014 / 052417 160 N--N I ,, N N-v 1H NMR (160) (yield 11 %): (400 MHz, CDCh) 5 8.59 (s, IH), 5.47 (s, 2H), 3.56 (d, J = 9.0 Hz, lH), 3.35 (s, 3H), 3.23 (d, J = 9.0 Hz, lH), 2.66 (t, J = 8.8 Hz, lH), 2.30-2.16 (m, lH), 2.15- 2.05 (m, IH), 1.98-1.89 (m, 2H), 1.84-1.41 (m, 14H), 1.40-1.11 (m, 9H), 0.72 (s, 3H). LCMS tR = 2.667 min in 4 min chromatography, 10-80AB, purity 100.0 %, MS ESI calcd. for C24H39N4O3 [M+Ht 431, found 413 [M+H-18t. Example 83. Preparation of Compound F5. C2 F3C H F3 TMSCF3 CsF Step 1 PCC DCM Step4 F3C H F4 t-BuOK,THF Step2 Br2 HBr MeOH F F H◊- F2 H F5 2) aq. NaOH. H2O2 Step3 Br Step 1. Preparation of Compound Fl. To a solution ofC2 (2 g, 6.28 mmol) in THF (30 mL) in a flask was added CsF (953 mg, 6.28 mmol) at O °C, then TMSCF3 (1.33 g, 9.42 mmol) was added dropwise. The reaction was allowed to warm to 25 °C and stirred for 2h. TLC(PE:EtOAc=3:l) showed the starting material was consumed completely. Then the reaction mixture was treated with 2M aq.HCI (10 mL) and stirred for 6h. The reaction was then diluted with H2O (30 mL) and extracted with EtOAc (30 mLx2). The combined organic layer was 151 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 washed with brine (20 mL), dried over Na2SO4 and concentrated to get the crude product which was purified by silica gel column (PE:EtOAc=50: 1 to 10: 1) to afford product Fl (I .1 g, 45.0% yield) as a yellow oil. 1H NMR (400 MHz, CDCh) 8 3.49(d, J = 8.0Hz, lH), 3.32-3.22(m, 4H), 2.46-2.39(111, IH), 2.10-1.71(m, SH), 1.68-1.10 (m, 14H), 0.85(s, 3H). Step 2. Preparation of Compound F2. To a solution of ethyltriphenylphosphonium bromide (5.19 g, 14.0 mmol) in THF (30 mL), was added t-BuOK (1.57 g, 14.0 mmol). The reaction mixture was heated to 60 °C for 1 h and Fl ( 1.1 g, 2.83 mmol) \Vas added to the mixture which was stirred at 60 °C for an additional 8 h. TLC (PE:EtOAc= 3: l) showed the reaction was complete. The reaction mixture was cooled, then diluted with H2O(30 mL) and extracted with EtOAc (30 mL x2). The combined organic layer was washed with brine (20 mL), dried over Na2SO4 and concentrated. The residue was purified by silica gel column (PE:EtOAc=l00:1 to 15:1) to afford product F2 (1 g, 88.6% yield) as a yellow oil. 1H NMR (400 MHz, CDCh) o 5.15-5.02(m, lH), 3.56(d, J= 8.0Hz, IH), 2.46-2.39(111, IH), 3.34(s, 3H), 3.29(d, J= 8.0Hz, IH), 2.43-1.80 (m, 7H), 1.58-1.10 (m, 19H), 0.90(s, 3H). Step 3. Preparation of Compound F3. To a solution ofF2 (1 g, 2.4911111101) in THF (15 mL) under N2 protection was added dropwise a solution ofBH3-Me2S (2.48 mL, 10 M) at O °C. The solution was stirred at 25 °C for 4h. TLC (PE / EtOAc = 3 / 1) showed the reaction was complete. After cooling to O °C, a solution ofNaOH (9.93 mL, 3M) was added very slowly, a large amount of gas released. After the addition was complete, H2O2 ( 4.53 mL, 33%) was added slowly and the inner temperature was maintained below 10 °C. The resulting solution was stirred at 25 °C for lh. The resulting solution was extract with EtOAc (20 mL x3). The combined organic solution was washed with saturated aqueous Na2S2O3 (20 mL x 3), brine (20 mL), dried over Na2SO4 and concentrated in vacuum to give the crude product (1 g) as yellow oil. The crude product was used for the next step without further purification. Step 4. Preparation of Compound F4. A mixture ofF3 (1.0 g, 2.38 nm101), PCC (0.767 g, 3.56 mmol) and silica gel (0.843 g, w / w = 1 / 1.1) in DCM (15 mL) was stirred at 25 °C for 2h, the reaction mixture color became brown. TLC (PE / EtOAc = 3 / 1) showed the reaction was complete. The solution was filtered and the filter cake was washed with DCM (20 mL). The combined filtrate was concentrated in vacuum. The residue was purified by silica gel column 152 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 eluted with PE:EtOAc = 15:1 to 8:1 to give F5 (800 mg, 80.6 %) as a white solid. MS ESI calcd. for C24:fLi1O4 [M+Hf 417, found 399 ( [M+H-18f). 1H NMR (400 MHz, CDCh) 8 3.52(d, .J=8.0Hz, IH), 3.33(s, 3H), 3.28(d, .J=8.0Hz, lH), 2.58-2.52 (m, lH), 2.20-1.60(m, 15H), 1.53- 1.I0(m, llH), 0.62(s, 3H). Step 5. Preparation of Compound F5. To a solution ofF4 (0.5 g, 1.20 mmol) and a catalytic amount of concentrated HBr (12.1 mg, 40% in water) in MeOH (15 mL) was added drop,vise dibromine (230 mg, 1.44 mmol) at 0 °C. The reaction mixture was stirred at 25 °C for 1 h. TLC (PE:EtOAc = 3:1) showed the reaction was complete. The reaction was quenched by saturated aqueous NaHCO3 and the pH was adjusted to 7-8. The reaction mixture was extracted with DCM (20 mLx2). The combined organic layer was washed with brine(20 mL), dried over Na2SO4 and concentrated to get the crude product F5 (500 mg) as a yellow oil. Example 84. Preparation of Compounds 161 and 162. 0 N ~=0- / 1 F Br HNC:C) N N N,- N~:,.,_ «-~~ F N F F F c:-.. F K2C03, acetone, 25 °c F F H 161 F5 162 To a solution of compound FS (150 mg, 0.302 mmol) in acetone (5 mL) was added K2CO3 (62.6 mg, 0.453 mmol) and 4,5-difluoro-2H-benzo[d][l,2,3]triazole (70.2 mg, 0.453 mmol). After stirring at 25 °C for 3 h, TLC (PE:EA=3: 1) showed the reaction was complete. The reaction mixture was filtered, and the filtrate was concentrated in vacuum to give the crude product (150 mg). The crude product was purified by prep. HPLC (HCl) to give compound 162 (18 mg, 10.4%) as a white solid and comp0tmd 161 (31 mg, 18%) as a white solid. 1H NMR (161) (yield 10.4%): (400 MHz, CDCh) 8 7.42-7.36 (m, IH), 7.09-7.06 (m, lH), 5.53- 5.35 (m, 2H), 3.50 (t, J= 9.2 Hz, lH), 3.34-3.29 (m, 4H), 2.71 (d,.J= 8.8 Hz, lH), 2.20-1.00 (m, 23H), 0.74 (s, 3H). LCM.S tR = 1.350 min in 2 min chromatography, I 0-S0AB, purity I 00%, MS ESI calcd. for C29H37FsN3O3 [M+Hf 570, found 570. 1HNMR(162)(yield 18%): (400MHz,CDCh)o7.68-7.64(m, lH), 7.37-7.29(m, lH),5.59- 5.49 (m, 2H), 3.51 (t, J= 7.6 Hz, IH), 3.34-3.29 (m, 4H), 2.69 (d, J= 8.8Hz, IH), 2.24-1.14 (m, 153 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 23H), 0.77 (s, 3H). LCMS tR = 1.398 min in 2 min chromatography, 10-80AB, purity 100%, MS ESI calcd. for C29H31FsN3Q3 [M+Ht 570, found 570. Example 85. Preparation of Compounds 163, 164, and 165. Br F5 + 163 F F 164 165 1H NMR (163) (yield 11 %): (400 MHz, CDCh) o 7.73 (t, J=5.6 Hz, lH), 7.09-7.06 (m, 2H), 5.48-5.38 (m, 2H), 3.88 (s, 3H), 3.50 (d, J=8.8Hz, lH), 3.31 (s, 3H), 3.28 (d, J=9.2 Hz, IH), 2.62 (t, J=8.8 Hz, lH), 2.17-1.11 (m, 23H), 0.74 (s, 3H). LCMS tR = 1.354 min in 2 min chromatography, 10-80AB, purity 100%, MS ESI calcd. for C3olii1f.,N3O4 [M+Ht 564, found 564. 1H NMR (164)(yield 11%): (400 MHz, CDCh) o 7.38 (d, J= 1.6 Hz, IH), 7.21 (d, J= 7.2 Hz, IH), 7.15 (dd, J1 = 7.2 Hz, 12 = 1.6 Hz, lH), 5.34 (s, 2H), 3.89 (s, 3H), 3.49 (d, J = 6.8 Hz, lH), 3.31 (s, 3H), 3.28 ( d, J = 6.8 Hz, lH), 2.66 (t, J = 7 .2 Hz, lH), 2.35-1.1 0 (m, 23H), 0.71 (s, 3H). LCMS tR = 0.964 min in 1.5 min chromatography, 5-95AB, purity 95%, MS ESI calcd. for C30H41F3N3O4 [M+Ht 564, found 564. 1HNMR(165)(yield 19%): (400MHz,CDCh)o7.91 (d,J=9.2Hz, lH), 7.01 (dd,J1 =9.2 Hz, J2 = 2.0 Hz, lH), 6.59 (d, J = 2.0 Hz, IH), 5.38-5.27 (m, 2H), 3.85 (s, 3H), 3.49 (d, J = 9.2 Hz, IH), 3.31 (s, 3H), 3.28 (d, J = 9.2 Hz, lH), 2.67 (t, J = 8.4 Hz, lH), 2.43 (hrs, IH), 2.30-1.05 (m, 23H), 0.71 (s, 3H). LCMS tR = 1.354 min in 2 min chromatography, 10-80AB, purity 100%, MS ESI calcd. for C30H41F3N3O4 [M+Ht 564, found 564. 154 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 Example 86. Preparation of Compound 166. F F Hd I 0 Br H F5 F F H PCT / US2014 / 052417 0 f~--~ N:::,.,. :::c--. ~ / 0 166 1H NMR (166) (yield 10.5%): (400 MHz, CDCh) & 7.73 (t, J= 5.6 Hz, IH), 7.09-7.06 (m, 2H), 5.48-5.38 (m, 2H), 3.88 (s, 3H), 3.50 (d, J = 8.8Hz, lH), 3.31 (s, 3H), 3.28 (d, J = 9.2 Hz, IH), 2.62 (t, J= 8.8 Hz, lH), 2.17-1.11 (m, 22H), 0.74 (s, 3H). LCMS tR = 1.354 min in 2 min chromatography, 10-80AB, purity 100%, MS ESI cakd. for C30H41F3N3O4 [M+Ht 564, found 564. Example 87. Preparation of Compounds 167, 168, and 169. F F . HcJ H + FS 167 F F + 168 169 1H NMR(167) (yield 1%): (400 MHz, CDC13) & 7.89-7.83 (m, 2H), 7.38 (t,J= 7.2 Hz, lH), 5.56-5.46 (m, 2H), 3.52 (d, J= 9.2 Hz, lH), 3.34 (s, lH), 3.30 (d, J= 9.2Hz, lH), 2.65 (t, J= 4.4Hz, lH), 2.26-1.07 (m, 25H), 0.76 (s, 3H). LCMS tR = 1.424 min in 2 min chromatography, 10-80AB, purity 99%, MS ESI calcd. for C29H38CW,N3O3 [M+Ht 568, found 568. 155 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 1H NMR (168) (yield 6%): (400 MHz, CDCh) & 8.01-7.99 (m, IH), 7.36-7.34 (m, 2H), 5.43- 5.31 (m, 2H), 3.50 (d, J=8.8 Hz, IH), 3.28-3.35 (m, 4H), 2.70 (t, J = 8.8Hz, IH), 2.24-1.13 (m, 23H), 0.73 (s, 3H). LCMS tR = 1.009 min in 1.5 min chromatography, 5-95AB, purity 97%, MS ESI calcd. for C29H3sClF3N3O3 [M+Hf 568, found 568. 1H NMR (169) (yield 8%): (400 MHz, CDCh) & 8.07 (s, lH), 7.45 (t, J = 7.2 Hz, IH), 7.29 (s, lH), 5.34-5.45 (m, 2H), 3.49 (d, J=8.8 Hz, lH), 3.28-3.32 (m, 4H), 2.70 (t, J = 9.2Hz, lH), 2.20-0.88 (m, 23H), 0.71 (s, 3H). LCMS tR = 0.995 min in 1.5 min chromatography, 5-95AB, purity 99%, MS ESI calcd. for C29H,8ClF3N3O3 [M+Ht 568, found 568. Example 88. Preparation of Compound Cl9. HBr MeOH Step3 C2 2. aq. NaOH, HP2 Step6 TBSCI, imidazole O MAD, MeMgBr --D-CM--► O H --1-olu-e-ne~- Step 1 Step 2 C14 ( 0 H C17 HO Step4 PCC DCM Step7 H C12 C15 C18 Ell NaH,THF Steps C13 ( 0 H C16 0 "i 0 H C19 Step 1. Preparation of Compound Cl2. To a solution of C2 (4 g, 13.14 mmol) in 15 mL CH2Ch was added lH-imidazole (2.68 g, 39.42 mmol) and tert-butylchlorodimethylsilane (2.97 g, 19.71 mmol) at 25 °C, the reaction was stirred at 25 °C for 16 h. The reaction mixture was 156 Date Rec;ue / Date Received 2024-04-12 Br WO 2015 / 027227 PCT / US2014 / 052417 filtered with 50 mL CH2Ch and evaporated in vacuo. The residue was purified by column chromatography on silical gel (PE:EtOAc= 50:1-30:1-20:1-15:l-10:1) to afford C12 (5 g, 90.87 % yield) as a white solid. 1H NMR: (400 MHz, CDCh) 8 3.81 (d, J = 8.0 Hz, lH), 3.58 (d, J = 8.0 Hz, IH), 2.62-2.55 (m, lH), 2.48-2.43 (m, IH), 2.40-2.28 (m, 3H), 2.26-2.07 (m, 2H), 1.87- 1.86 (m, lH), 1.84-1.75 (m, 4 H), 1.56-1.27 (m, IOH), 0.87 (s, 12H), 0.042(s, 6H). Step 2. Preparation of Compound C13. To a solution of C12 (15.79g, 71.64 mmol) in 30 mL toluene was added a solution of A1Me3(17.91 mL, 3eq) dropwise at O 0C. After 1 h, a solution of (5R,8R,9S, 1 OR, l 3S,14S)-1 0-(((tert-butyldimethylsilyl)oxy)methyl)-13-methyldodecahydroIH- cyclopenta[a]phenanthrene-3, l 7(2H,4H)-dione (5 g, 11.94 mmol) in toluene (40 mL) was added drop wise at -78 °C, the reaction mixture was stirred at -78 °C for 1 h and then a solution of MeMgBr ( 11.94 mL, 3eq) was added drop wise to the mixture at -78 °C which was stirred at - 78 °C for another 2 h. After TLC (PE:EtOAc = 3: 1 ) showed the starting material was consumed completely, the reaction mixture was quenched with aq.NH4C1 (15 mL), filtered and washed with 500 mL EtOAc. The organic layer was extracted with 300 mL EtOAc, washed with brine and concentrated. The residue was purified by column chromatograph on silica gel (PE:EA=l00- 50: l-20:1-10:1-4:1) to give C13 (5 g, 96.3%) as a white solid. 1H NMR (400 MHz, CDCh), 8 3.76 (d, J = 8.0 Hz, lH), 3.40 (d, J = 8.0 Hz, IH), 2.44-2.39 (m, lH), 2.09-1.70 (m, 6H), 1.61- 1.18 (m, 19H), 0.89 (s, 12H), 0.04 (s, 6H). Ste1> 3. Preparation of Compound C14. To a solution ofC13 (7 g, 16.1 mmol) in 70 mL MeOH was added a solution ofHBr (6.5 g, 32.2 mmol, 40% in water). The reaction mixture was stirred at 25 °C for 0.7 h. After TLC (PE:EtOAc = 3: 1) showed the starting material was consumed completely, the reaction mixture was quenched with sat.aq.NaHCO3 (200 mL) and extracted with 500 mL EtOAc, washed with brine ( I 00 mL) and concentrated to give product C14 (5.6 g, crude) as a white solid. Step 4. Preparation of Compound C15. To a solution of PPh3EtBr (51.8 g, 140 mmol) in THF (40 mL) was added a solution oft-BuOK (15.7 g, 140 mmol) in THF (40 mL) at 0 °C. After stirring at 60 °C for 1 h, a solution of compound C14 (9 g, 28.0 mmol) in THF (40 mL) was added dropwise at 60 °C. Then the reaction mixture was stirred at the same temperatrue for 8 h. TLC (PE / EtOAc = 3 / I) showed the reaction was completed, and a main product was found with lower polarity. The reaction mixture was extracted with EtOAc (300 mL) for three times. The 157 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 organic layer was washed with brine (100 mL), dried over Na2SO4 and concentrated in vacuum to give the crude product. The crude product was purified by a silica gel column (PE:EA=5: 1) to give compound C15 (5.0 g, 53.5%) as a pale yellow oil. 1H NMR (400 MHz, CDCb) o 5.15- 5.05 (1.11, lH), 3.94 (d,J= 10.8Hz, lH), 3.56 (d,J= 10.8Hz, lH), 2.40-2.12 (m, 3H), 2.0ll. 7l(m, 3H), l.69-l.12(m, 24H), 0.85(s, 3H). Step 5. Preparation of Compound C16. To a solution of C15 (500 mg, 1.50 11111101) in THF (15 mL) in a flask under N2 protection was added NaH (171 mg, 4.5 mmol, 60% in oil) in portions. The reaction mixture was stirred for 10 min. Then iodoethane (701 mg, 4.5 mmol) was added. The reaction mixture was heated and stirred at 50 °C for another 2h. TLC(PE:EA=3:l) showed the reaction was complete, and a main product was found with lower polarity. The reaction was queched with aq.N~Cl(lO ml), extracted with EtOAc(20mLx2). The combined organic layers were washed with aq. NaCl (20 mL) and dried over Na2SO4, then concentrated. The residue was purified by column chromatography on silica gel (PE:EtOAc= 15: 1 to 8: 1) to afford the product C16 (500 mg, 91.9 % yield) as a yellow oil. 1H NMR (400 MHz, CDCb) o 5.12-5.09(m, lH), 3.60(d, J = 9.2 Hz, lH), 3.48-3.42(111, 2H), 3.23(d, J = 9.2 Hz, lH), 2.38- 2.12(m, 3H), l.95-1.72(m, 3H), 1.65-1.10 (m, 26H), 0.85(s, 3H). Step 6. Preparation of Compound C17. To a solution ofC16 (500 mg, 1.38 mmol) in THF (15 mL) was added dropwise a solution ofBH3-Me2S (1.38 mL, 10 M) at 0°C. The solution was stirred at 25 °C for 4h. TLC (PE:EtOAc = 3:1) showed the reaction was almost complete, and a main product was found with higher polarity. After cooling to 0°C, a solution of NaOH (5.5 mL, 3M) was added very slowly. After the addition was complete, H2O2 (2.51 mL, 33%) was added slowly and the inner temperature was maintained below 10 °C. The resulting solution was stirred at 25 °C for 2h. The resulting solution was extract with EtOAc (20 mL x3). The combined organic solution was washed with saturated aqueous Na2S2O3 (30 mL x 3), brine (30 mL), dried over Na2SO4 and concentrated in vacuo to give the crude product (500 mg) as a yellow oil. The crude product was used for the next step \Vithout further purification. Step 7. Preparation of Compound C18. A suspension ofC17 (500 mg, 1.32 rnmol), PCC (426 mg, 1.98 mmol) and silica gel (469 mg, w / w = 1 / 1.1) in DCM (15 mL) was stirred at 30 °C for 2h, the reaction mixture color became bmwn. TLC (PE / EtOAc = 3 / 1) showed the reaction was 158 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 complete, and a main product was found with lower polarity. The solution was filtered and the filter cake was washed with DCM (20 mL). The combined filtrate was concentrated in vacuo. The residue was purified by column chromatography on silica gel eluted with PE / EtOAc = 15 / 1 to 5 / 1 to give C18 (400 mg, 80.3 %) as a white solid. MS ESI calcd. for C24H40O3 [M+Ht 377, found 359([M+H-18t) . 1H NMR (400 MHz, CDCh) 8 3.58 (d, J = 9.2Hz, IH), 3.49-3.42 (m, 2H), 3.24 (d, J = 9.2Hz, lH), 2.56-2.51 (111, lH), 2.18-1.65 (m, 12H), 1.60-1.10(111, 19H), 0.6l(s, 3H). Step 8. Preparation of Compound C19. To a solution of C18 (400 mg, l.06 11111101) and a catalytic amount of concentrated HBr (10.7 mg, 40% in water) in MeOH (15 mL) was added dropwise dibromine (254 mg, 1.59 mmol) at O °C. The reaction mixture was stirred at 25 °C for I h. TLC (PE:EtOAc = 3:1) showed the reaction was complete, and a main product was found with lower polarity. The reaction was quenched by saturated aqueous NaHCO3 and the pH was adjusted to 7~8. The reaction mixture was extracted with DCM (20 mLx2). The combined organic layer was washed with brine (20 mL), dried over Na2SO4 and concentrated to get the crude product Cl9 (400 mg, 82.8 % yield) as yellow oil. Example 89. Preparation of Compound 170. l 0 Br N HN°' CN l 0 H HO H F5 170 To a solution of compound F5 (150 mg, 0.329 mmol) in acetone (5 mL) was added K2CO3 (68.1 mg, 0.493 mmol) and IH-pyrazole-4-carbonitrile (45.8 mg, 0.493 mmol). After stirring at 25 °C for 3 h, LCMS showed the reaction was complete. The reaction mixture was filtered, and the filtrate was concentrated in vacuum to give the crude product (150 mg). The crude product was purified by prep. HPLC (HCl) to give the desired product 170 (13 mg, 8.41 %) as white solid. 159 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 1H NMR (170) (yield 8.4%): (400 MHz, CDCh) o 7.86(s,1H), 7.81 (s, IH), 5.03-4.87 (m, 2H), 3.54(d, J= 9.2Hz, IH), 3.68-3.41 (m, 2H), 3.24 (d, J= 9.2Hz,1H), 2.59 (t, J= 9.2Hz, lH), 2.21- 1.15 (m, 29H), 0.65 (s, 3H). LCMS tR = 0.949 min in 1.5 min chromatography, 5-95AB, purity 98.6%, MS ESI calcd. for C2s!Li2N3O3 [M+Ht 467, found 450[M+H-18t. Example 90. Alternative Preparation of Compound A21. 0 E4 NaH,Etl THF Step 3 A20 MeMgBr THF Step 1 A18 HBr,Br2 MeOH Step 6 EtPPh3Br t-BuOK,THF Step2 E16 1) BH3Me2S 2) 10% aq. NaOH, H2O2 Step 4 Br A21 E17 HO Pee.., DCM Steps A19 Step 1. Preparation of Compound E16. To a solution of((5S,8R,9S,lOR,l3S,14S)-13-methyl- 3,17-dioxohexadecahydro-lH- cyclopenta[a]phenanthren-10-yl)methyl acetate (E4, 5 g, 14.4 mmol) in THF (50 mL) was added MeMgBr (I 5 mL, 3M in ether, 450 mmol) dropwise to control inner temperature below -70 oC. The mixture was then stirred for I hour at -78 oC.. TLC showed the reaction was completed. To the mixture was added a solution of NH4Cl (6 g) in 160 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 water (30 mL) and inner temperature was raised to -20 oC. The mixture was then wanned to 20 oC. Organic layer was separated. The aqueous phase was extracted with EtOAc (50 mL). The combined organic layer was dried over Na2SO4, concentrated under vacuum and purified by cloumn chromatography (PE: EtOAc =6:1 to 3:1) to give ((3R,5S,8R,9S,IOR,13S,14S) -3- hydroxy-3, 13-dimethyl-17-oxohexadecahydro-IH-cyclopenta[ a ]phenanthren-10-yl)methyl acetate (E16, 2.4 g, 46%) and ((3S,5S,8R,9S,IOR,13S,14S)-3-hydroxy-3,13- dimethyl-17- oxohexadecahydro-lH-cyclopenta[a]phenanthren-10-yl)methyl acetate (1 g, 19%) as white solid. 1H NMR (400 MHz, CDCh) J 4.29 (d, J = 12.1 Hz, lH), 4.13 (d, J = 12.1 Hz, lH), 2.48-2.35 (m, lH), 2.11-1.85 (m, 7H), l.85-1.59 (m, 6H), l.55-1.22 (m, llH), 1.09 - 0.74 (m, 7H). Step 2. Preparation of Compound E17. To a suspension of PPh3EtBr (4.61 g, 12.4 mmo1) in THF (10 mL) was added a solution of t-BuOK (1.86 g, 16.6 mmol) in THF (20 mL) at 20 oC. The color of the suspension was turned to dark red. After stirring at 60 °C for 1 h, a solution of ((3R,5S,8R,9S, 1 OR, 13S, 14S) -3-hydroxy-3,13-dimethyl-l 7-oxohexadecahydro-l Hcyclopenta[ a]phenanthren-10-yl)methyl acetate (E16, 1.5 g, 4.14 mmol) in THF (20 mL) was added dropwise at 60 °C. Then the reaction mixture was stirred at 60 °C for 16 h. TLC showed the reaction was complete. To the reaction mixture was added NH4Cl (50 rnL, sat. aq.). The color of the mixture was turned to light yellow. The organic layer was separated. The aqueous phase was extracted with EtOAc (50 mL). The combined organic layer was concentrated under vacuum purified by column chromatography on silica gel (PE: EtOAc = 10: 1 to 4: 1) to give (3R,5S,8S,9S, 1 OR, 13S, 14S)-l 7-ethylidene-l 0-(hydroxymethyl)-3, 13-dimethylhexadecahydrol H-cyclopenta[a]phenanthren-3-ol (E17, 1.0 g, 72.6%) as white solid. 1H NMR (400 MHz, CDCb) <> 5.19-5.08 (m, lH), 3.93 (d, J = I 1.5 Hz, IH), 3.74(d,J=11.5 Hz, lH), 2.44-2.33 (m, lH), 2.32-2.13 (m, 2H), 2.12-2.04 (m, lH), 1.87 - 1.71 (m, 2H), 1.69 - 1.43 (m, 12H), 1.38 - 1.08 (m, llH), 1.07 - 0.74 (m, 6H) Step 3. Preparation of Compound A18. To a solution ofE17 (0.8 g, 2.4 mmol) in THF (10 mL) was added sodium hydride (475 mg, 11.9 mmol) in portions and iodoethane (1.85 g, 11.9 mmol). TI1e mixture was stirred at 50°C for 12 hours. The reaction mixture was quenched with water, extracted with EtOAc ( 10 mL *2). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel (PE:EA = 50:1) to give A18 (0.5 g, 57.5%) as colorless oil. 1H NMR (400 MHz, CDCh) ,5 161 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 5.13-5.08 (m, IH), 3.52 (d, J = 9.6 Hz, IH), 3.43-3.38 (m, 3H), 2.38-2.32 (m, IH), 2.25-2.12 (m, 2H), 2.06-2.01 (m, lH), 1.74-1.57 (m, 2H), l.56-1.39 (m, 6H), 1.31-1.27 (m, 2H), 1.24-1.16 (m, l IH), 1.14-1.07 (m, 2H), 1.05-0.91 (m, 2H), 0.88 (s, 3H), 0.87-0.74 (m, 3H). Step 4. Preparation of Compound Al9. To a solution of Al8 (0.5 g, 1.38 mmol) in THF (5 mL) at 0°C was added BH3-Me2S (0.69 mL, 6.9 mmol) dropwise. The solution was stirred at 30 °C for 2 h. TLC (PE / EtOAc = 5 / 1) showed the reaction was completed. After cooling to O °C, an aqueous NaOH (5.51 g, 10% in water) was added very slowly. After the addition was completed, H2O2 (1.56 g, 30%) was added slowly and the inner temperature was maintained below 10°C. The resulting solution was stirred at room temperature for lh. Wl1ite solid was formed. To the mixture was added EtOAc (5 mL) and filtered. The filter cake was washed with EtOAc (5 mL). The combined organic layer was separated, washed with Na2S2O3 (5 mL, 20%, aq.), dried over Na2SO4 and concentrated in vacuum to give Al9 (0.4 g, purity: 78%, yield: 59.7%) as colorless oil which was used directly without further purification. LCMS tR = 1.085 min in 2 min chromatography, 30-90AB, purity 77.6%, MS ESI calcd. for C24H42O3 [M+Ht379, found 361 ([M+H-lSt). Step 5. Preparation of Compound A20. To a solution of Al9 (0.4 g, 0.824 mmol, purity: 78%) in dichloromethane (5 mL) was added silica gel (1 g) and PCC (0.885 g, 4.11 mmol). The suspension was stirred at 30°C for 16 hours. TLC (PE:EA = 5: 1) showed the reaction was consumed completely. The reaction mixture was filtered, and the filtrate was concentrated. The residue was purified by column chromatography on silica gel (PE:EA=l 0: 1) to give A20 (0.2 g, 64.4%) as light yellow oil. 1H NMR (400 MHz, CDCh) LJ 3.50 (d, J= 10.0 Hz, lH), 3.42-3.37 (m, 3H), 2.53 (t, J = 8.8 Hz, lH), 2.20-2.15 (m, lH), 2.11 (s, 3H), 2.07-1.97 (m, 2H), 1.73-1.64 (m, 4H), 1.50-1.47 (m, 2H), l.37-1.25 (m, 6H), 1.21- 1.14 (m, 9H), 1.12-0.75 (m, SH), 0.61 (s, 3H). LCMS tR "" 1.124 min in 2 min chrornatography, 30-90 / \B, purity 100%, MS ESI calcd. for C24H4li(J:i [M+Hr 377, found 359 (Uvl+H-l St). Step 6. Preparation of Compound A21. To a solution of A20 (0.2 g, 0.531 mmol) in methanol (2 mL) was added HBr (8.93 mg, 0.053 mmol, 48% in water) and Br2 (127 mg, 0.796 mmol). The mixhtre was stirred at 30°C for 2 hours. The reaction mixh1re was quenched with aqueous NaHCO3 to adjust the pH about 8. The mixture was poured to water (10 mL) and extracted with 162 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 EtOAc ( 10 mL *2). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated to give A.21 (0.2 g, 82.6%) as a light yellow solid which was used without further purification. LCMS tR = 1.184 min in 2 min chromatography, 30-90AB, purity 100%, MS ESI calcd. for C24H39BrO3 [M+Hr 455, found 437 ([M+H-18t). Example 91. Preparation of Compound 171. Br A21 H~~CN Nd BB-27 171 To a solution of A21 (90 mg, 0.197 mmol) in acetone (2 mL) was added potassium carbonate (67.9 mg, 0.492 mmol) and IH-pyrazole-4-carbonitrile (27.4 mg, 0.295 mmol). The suspension was stirred at 40°C for 12 hours. The reaction mixture was cooled and filtered, and the filtrate was concentrated. The residue was purified by prep. HPLC to give 171 (22 mg, 23.8%) as a white solid. 1H NMR (400 MHz, CDCb) 5 7.85 (s, lH), 7.81 (s, lH), 5.02 (d, J= 18.0 Hz, lH), 4.89 (m, J = 18.0 Hz, IH), 3.51 (d, J = 9.2 Hz, lH), 3.42-3.37 (m, 3H), 2.60 (t, J = 9.2 Hz, IH), 2.25-2.17 (m, lH), 2.06-1.99 (m, 2H), 1.75-1.69 (m, 4H), 1.54-1.50 (m, 3H), 1.46-0.81 (m, 19H), 0.67 (s, 3H). LCMS tR = 1.109 min in 2 min chromatography, 30-90AB, purity 100%, MS ESI calcd. for C28H41N3O3 [l\HHt 468, found 490 ([M+Nat). Example 92. Preparation of Compounds 172 and 173. 173 A21 163 Date Rec;ue / Date Received 2024-04-12 WO 2015 / 027227 PCT / US2014 / 052417 To a solutin of 2-bromo-l-((3R,5S,8S,9S,10R,13S, l 4S,17S)-l 0-(ethoxymethyl)-3-hydroxy- 3, 13-dimethylhexadecahydro-1 H-cyclopenta[ a ]phenanthren-17-yl)ethanone ( 90 mg, 0 .197 mmol) in acetone (2 mL) was added potassimn carbonate (67.9 mg, 0.492 mmol) and 4,5- difluoro-2H-benzo[d][l,2,3]triazole (45.7 mg, 0.295 mmol). The mixture was stirred at 50 °C for 16 hours. The mixture was cooled, filtered and concentrated. The residue was purified by prepHPLC to give 2-( 4 ,5-difluoro-2H-benzo[ d][ 1,2,3 ]triazol-2-yl)-1-((3R,5S, 8S,9S, l0R, 13S, l 4S,17S)-l 0-( ethoxymethyl)-3~hydroxy-3, l 3-dimethylhexadecahydro-IHcyclopenta[ a]phenanthren-17-yl)ethanone (172, 10 mg, 9.28%, purity: 97%) as light yellow solid and 2-( 4,5-difluoro-1H-benzo[dj[1,2,3]triazol-l-yl)-1-((3R,5S,8S,9S,1 OR, 13S, l 4S, 17S )-10- ( ethoxymethyl)-3-hydroxy-3, 13-dimethylhexadecahydro-lH-cyclopenta[ a ]phenanthren-17- yl)ethanone (173, 15 mg, 14.0%, purity: 98%) as light yellow solid. 1H NMR (172): (400 MHz, CDCh) J 7.63 (dd, .J= 3.2 Hz, 8.8 Hz, lH), 7.31-7.24 (m, 2H), 5.57-5.46 (m, 2H), 3.53 (d, .J = 9.6 Hz, lH), 3.43-3.40 (m, 3H), 2.66 (t, .J = 8.8 Hz, lH), 2.27- 2.03 (m, 3H), 1.79-1.69 (m, 4H), 1.65-1.57 (m, 4H), 1.32-1.26 (m, 6H), 1.22-1.12 (m, 8H), 1.09- 0.82 (m, 4H), 0.76 (s, 3H). LCMS tR = 1.066 min in 1.5 min chromatography, 5-95AB, purity 97%, MS ESI calcd. for C30H42F2N3O3 [M+Hf 530, found 512([M+H-18f). 1H NMR (173): (400 MHz, CDCh) J 7.40-7.33 (m, lH), 7.05 (d, J = 7.2 Hz, 1H), 5.46-5.35 (m, 2H), 3.52 (d, .J = 10.0 Hz, lH), 3.43-3.38 (m, 3H), 2.71 (t, .J = 8.4 Hz, lH), 2.24-2.03 (m, 3H), 1.75-1.69 (m, 4H), 1.62-1.53 (m, 4H), 1.32-1.16 (m, 14H), 1.13-0.83 (m, 4H), 0.72 (s, 3H). LCMS tR = 1.037 min in 1.5 min chromatography, 5-95AB, purity 98%, MS ESI calcd. for C30H42F2N3O3 [M+Hf 530, found 530([M+H...

Claims

92460299 CLAIMS:

1. A compound of Formula (A13): (A13), or a pharmaceutically acceptable salt thereof.

2. The compound:

3. A pharmaceutically acceptable salt of the compound:

4. A compound of Formula (A16): (A16), or a pharmaceutically acceptable salt thereof. 176 Date Rec;ue / Date Received 2024-04-12 92460299 5. The compound:

6. A pharmaceutically acceptable salt of the compound: 177 Date Rec;ue / Date Received 2024-04-12