Modulators of hydroxysteroid 17b dehydrogenase type 10

Abstract

Provided herein are compounds having a structure of Formula (I) or as disclosed in Table 1 herein, and methods of using the disclosed compounds to modulate 17P-HSD10 activity, such as to inhibit 173-HSD10 activity. For example, the compounds of Formula I and Table 1 as disclosed herein are useful for methods of treating diseases and disorders including, without limitation, neurological diseases and disorders such as Alzheimer's disease, and cancer. Also provided are methods of synthesizing the compounds disclosed herein, including compounds having a structure of Formula (I), Compound 1, and compounds of Table A disclosed herein.

Description

MODULATORS OF HYDROXYSTEROID 17 DEHYDROGENASE TYPE 10 STATEMENT OF GOVERNMENT INTEREST

[0001] This invention was made with government support under Grant No. T32 AG076407 awarded by the National Institutes of Health. The government has certain rights in the invention. BACKGROUND

[0002] The present invention in its various embodiments describes novel chemical compositions, methods of modulating hydroxysteroid 17β dehydrogenase type 10 (17β-HSD10) activity, and / or methods of treating a disorder in a subject associated with aberant 17β-HSD10 activity.

[0003] 17β-HSD10 is a mitochondrial enzyme implicated in multiple disease types, including, but not limited to, Alzheimer’s disease and cancer. The enzyme is a multifunctional nicotinamide adenine dinucleotide (NAD+)- dependent homo tetramer protein complex with many reported roles, including the degradation of isoleucine, metabolism of neuro-steroids, and the processing of mitochondrial tRNA transcripts. The crystal structure of 17β- HSD10 indicates a flexible active center that is directed outwards, resulting in its wide substrate specificity encompassing steroids, simple alcohols, faty acids, and amino acid metabolites.

[0004] Modulating the activity of 17β-HSD10 may be helpful in the treatment of diseases such as neurological disorders and diseases (e.g., Alzheimer’s disease) and cancer. SUMMARY

[0005] Provided herein are compounds, and pharmaceuticaly acceptable salts thereof, having the structure of Formula (I): (I), wherein R is OR1, SR1, NRaRb, S(O)R1, S(O2)R1, or P(O2)R1; R1 is C1-6alkyl, C2-6alkenyl,5- to 10 membered heterocycloalkyl comprising one to three heteroatoms selected from O, N, and S, or 5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S; and Ra and Rb are individualy H, C1-6alkyl, C2-6alkenyl, C(O)OC1-6alkyl, C3-10cycloalkyl, C6-10aryl, 5- to 10 membered heterocycloalkyl comprising one to three heteroatoms selected from O, N, and S, or 5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S. Also provided are compounds and salts having a structure as shown in Table A.

[0006] Also provided are methods of modulating 17β-HSD10 activity, comprising contacting a cel with a therapeuticaly efective amount of a compound or salt disclosed herein (e.g., a compound of Formula (I), acompound as shown in Table A, or a pharmaceuticaly acceptable salt thereof). Further provided are methods of modulating 17β-HSD10 activity in a subject, comprising administering to the subject a therapeuticaly efective A,disclosed herein (e.g., a compound of Formula (I), a compound as shown in Table A, or a pharmaceuticaly acceptable salt thereof). In various aspects, the disease or disorder is associated with aberant 17β-HSD10 activity, for example, a neurological disease or disorder (such as Alzheimer’s disease), or cancer.

[0008] Also provided are synthesizing a compound of Formula (I), or a salt thereof: N, (3-comprising admixing Compound 11 and N,N-dimethylformamide dimethyl acetal (DMF-DMA) under conditions suficient to form Compound 12. In various aspects, the compound of Formula (I) has a structure of Formula (I), or as shown in Table A. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] FIG.1 shows docking of Compound 1 into the X-ray structure of 17β-HSD10 (PDB ID: 1U7T). In the structure of 1, dark doted lines indicated predicted strong hydrogen bonds. Light doted lines indicate weak hydrogen bonds.

[0010] FIG 2 shows the results of a protein thermal shift assay showing concentration-dependent decrease of the 17β-HSD10 melting temperature with increasing concentration of 1.

[0011] FIGs 3A, 3B, 3C, 3D, and 3E show an assessment of cel viability on treatment of cel lines A) SH-SY5Y, B) LNCaP, C) PANC-1, and D) multiple additional cel lines with varying concentrations of 1. Unpaired T-test; 95% confidence interval; **, P < 0.0001.

[0012] FIGs 4A, and 4B show A) Co-treatment of Aβ and 1 in SH-SY5Y cels for 48h incubation. B) Pre-treatment of 1 in SH-SY5Y cels for 24h folowed by addition of Aβ (no wash step). Data represents mean of n=3 ±SEM experiments run in triplicate. Unpaired t-test; 95% confidence interval; *, p < 0.05; * p < 0.001, n=3.

[0013] FIGs 5A, 5B, 5C, and 5D show pre-treatment of Aβ and 14b, 14c, 14d, and 14e, respectively, for A, B, C, and D in SH-SY5Y cels for 24h, folowed by addition of Aβ (no wash step). Data represents mean of n=3 ±SEM experiments run in triplicate. Unpaired t-test; 95% confidence interval; *, p < 0.05. DETAILED DESCRIPTION

[0014] Alzheimer’s Disease (AD) is estimated to afect over 55 milion people across the world. Smal molecule treatment options are limited to symptom management with no impact on disease progression. The need for new protein targets and smal molecule hit compounds is unmet and urgent. Hydroxysteroid 17-beta dehydrogenase 10 (17β-HSD10) is a mitochondrial enzyme known to bind amyloid beta (Aβ), a halmark of AD, and potentiate its toxicity to neurons. Identification of smal molecules capable of interacting with 17β-HSD10 may drive drug discovery eforts for AD. The screening compound BCC0100281 (also caled “Compound 1” herein), was previously identified as an inhibitor of 17β-HSD10:

[0015] Herein is provided the first synthetic access to Compound 1 folowing a convergent pathway starting from simple heterocycle building blocks. The compound was found to be toxic to ‘neuron-like’ cels. However, assay of synthetic intermediates identified novel scafolds with efect to rescue amyloid beta-induced cytotoxicity, showcasing the power of organic synthesis and medicinal chemistry to optimize hit compounds to identify potential leads.

[0016] Hydroxysteroid 17β dehydrogenase type 10 (17β-HSD10) is a mitochondrial enzyme implicated in multiple disease types including, but not limited to; Alzheimer’s disease (AD),cancer, and other like diseases and disorders. The enzyme is a multifunctional nicotinamide adenine dinucleotide (NAD+)-dependent homo tetramer protein complex with many reported roles including the degradation of isoleucine, metabolism of neurosteroids and the processing of mitochondrial tRNA transcripts.1-4 The crystal structure of 17β-HSD10 indicates a flexible active center that is directed outwards resulting in its wide substrate specificity encompassing steroids, simple alcohols, faty acids and amino acid metabolites.5, 6

[0017] Amyloid beta (Aβ), one of the three pathological halmarks of AD, has been shown to bind 17β-HSD10, leading to potentiation of Aβ toxicity, increased reactive oxygen species and mitochondrial dysfunction.7-10 Furthermore, 17β-HSD10 modulates the level of multiple substrates in the brain including; acetoacetyl-CoA and estradiol, which are essential for neuronal survival. Additionaly, 17β-HSD10 acts as a key energy regulator and plays a role in regulating brain metabolic homeostasis.11-13 Levels of 17β-HSD10 have been found to be elevated in regions of the brain afected by AD pathology.14 The 17β-HSD10-Aβ protein-protein interaction in both AD patients and mouse models has been linked to disturbances in the balance of estradiol, peroxiredoxin-2, alopregnanolone, and endophilin-1 in the brain. Estradiol exhibits neuroprotective efects within the brain through many routes.11, 15-21 The reduction of estradiol due to the 17β-HSD10-Aβ interaction increases brain macrophage reactivity, Aβ accumulation, and increases the risk of hypermetabolism in the brain. Peroxiredixin-2, an antioxidant, is found to be elevated in AD.22 However, this protein is inactivated due to the kinase CDK5 phosphorylation.23 Endophilin-1 is responsible for synaptic vesicle endocytosis.24 However, elevated levels of endophilin-1 in AD result in neuroinflammation, another pathological halmark of AD, and apoptosis due to the activation of c-Jun N-terminal kinase.25, 26 Therefore, inhibition of the Aβ-17β-HSD10 protein-protein interaction through a smal molecule intervention or the direct inhibition of the enzyme has the potential to be exploited as a therapy for AD.27-30

[0018] Testosterone and 5α-dihydrotestosterone (DHT) activate the androgen receptor (AR), which contributes to the survival and growth of prostate cancer cels.31 It is suggested that 17β-HSD10 may play a role in the survival of prostate cancer cels in those patients who are undergoing androgen deprivation therapy or are post-castration. This is due to 17β-HSD10 possessing the ability to catalyze the reaction of 5α-androstane-3α,17β-diol into DHT, thus activating the AR in the absence of testosterone.32, 33 Therefore, the elevated levels of 17β-HSD10 found in prostate cancer corelate with increased DHT levels and proliferation.33-36 Additionaly the upregulation of the 17β-HSD10 gene corelates to poor treatment response in osteosarcoma.34 Thus, the targeted inhibition of 17β-HSD10 could provide new treatment options for prostate and other cancers where the enzyme is upregulated. Hence, identifying diverse molecules with the ability to modulate the activity of 17β-HSD10 is of interest to address multiple areas of unmet medical need.Compounds of the Disclosure

[0019] Provided herein are compounds and pharmaceuticaly acceptable salts thereof, having a structure of Formula (I):R is OR1, SR1, NRaRb, S(O)R1, S(O2)R1, or P(O2)R1; R1 is C1-6alkyl, C2-6alkenyl, C3-10cycloalkyl, C6-10aryl, 5- to 10 membered heterocycloalkyl comprising one to three heteroatoms selected from O, N, and S, or 5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S; and Ra and Rb are individualy H, C1-6alkyl, C2-6alkenyl, C(O)OC1-6alkyl, C3-10cycloalkyl, C6-10aryl, 5- to 10 membered heterocycloalkyl comprising one to three heteroatoms selected from O, N, and S, or 5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S.

[0020] In some cases, R is SMe, OMe, NH2, NHMe, or NHBoc, SR1, S(O)R1, S(O2)R1, OR1, NHR1, NR1R2, or P(O2)R1; and R1 and R2 is any alkyl, aryl, akenyl, alkenyl, cyclic or heterocyclic substituent.

[0021] In some cases, R is OR1, SR1, or NRaRb. In some cases, R is OR1 or SR1. In some cases, R is OR1. In some cases, R is SR1. In some cases, R1 is C1-6alkyl, C2-6alkenyl, C3-10cycloalkyl, C6-10aryl, 5- to 10 membered heterocycloalkyl comprising one to three heteroatoms selected from O, N, and S, or 5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S. In some cases, R1 is C1-6alkyl. In some cases, R1 is methyl. In some cases, R is NRaRb. In some cases, Ra is H. In some cases, Rb is H. In some cases, Rb is C1- 6alkyl or C(O)OC1-6alkyl. In some cases, Rb is C1-6alkyl. In some cases, Rb is methyl. In some cases, Rb is C(O)OC1-6alkyl. In some cases, Rb is C(O)O-t-butyl. In some cases, R is SMe, OMe, NH2, NHMe, or NHBoc. In some cases, R is OMe or SMe. In some cases, R is OMe. In some cases, R is SMe. In some cases, R is NH2, NHMe, or NHBoc. In some cases, R is NH2. In some cases, R is NHMe. In some cases, R is NHBoc.

[0022] Specific compounds contemplated include those listed in Table A, and pharmaceuticaly acceptable salts thereof:Table A Compound Structure 14a 14b 14c 14d 14e

[0023] In some cases, the compound or salt has the structure of Compound 14c or 14d. In some cases, the compound or salt has the structure of Compound 14c. In some cases, the compound or salt has the structure of Compound 14d.

[0024] As used herein, the term “alkyl” refers to straight chained and branched saturated hydrocarbon groups containing one to six carbon atoms. The term Cn means the alkyl group has “n” carbon atoms. For example, C6 alkyl refers to an alkyl group that has 6 carbon atoms. “C1-C6alkyl” refers to an alkyl group having a number ofcarbon atoms encompassing the entire range (e.g., 1 to 6 carbon atoms), as wel as al subgroups (e.g., 1-6, 2-6, 1- 5, 3-6, 1, 2, 3, 4, 5, and 6 carbon atoms). Nonlimiting examples of alkyl groups include, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl (2-methylpropyl), t-butyl (1,1-dimethylethyl), and 3-methylpentyl. Unless otherwise indicated, an alkyl group can be an unsubstituted alkyl group or a substituted alkyl group.

[0025] The term “alkenyl” used herein refers to an unsaturated aliphatic group analogous in length and possible substitution to an alkyl group described above, but that contains at least one double bond. For example, the term “alkenyl” includes straight chain alkenyl groups (e.g., ethenyl, propenyl, butenyl, pentenyl, hexenyl), and branched alkenyl groups (e.g., prenyl and reverse prenyl). A straight chain or branched alkenyl group can have six or fewer carbon atoms in its backbone (e.g., C2-C6 for straight chain, C3-C6 for branched chain). The term “C2-6alkenyl” includes chains having a number of carbon atoms encompassing the entire range (e.g., 2 to 6 carbon atoms), as wel as al subgroups (e.g., 2-6, 2-5, 2-4, 3-6, 2, 3, 4, 5, and 6 carbon atoms). Unless otherwise indicated, an alkenyl group can be an unsubstituted alkenyl group or a substituted alkenyl group.

[0026] The term "cycloalkyl" refers to a non-aromatic monocyclic, fused, bridged or spiro ring system whose ring atoms are carbon and which can be saturated or have one or more units of unsaturation. The cycloalkyl can have three to ten ring carbon atoms. In some embodiments, the number of carbon atoms is 5 to 6. In some embodiments, the number of carbon atoms is 6. "Fused" bicyclic ring systems comprise two rings which share two adjoining ring atoms. Bridged bicyclic group comprise two rings which share three or four adjacent ring atoms. Spiro bicyclic ring systems share one ring atom. Cycloalkyl groups can include cycloalkenyl groups. Specific examples include, but are not limited to, cyclohexyl, cyclopentyl, cyclobutyl, and cyclopropyl.

[0027] The term "heterocycloalkyl" as used herein refers to a non-aromatic monocyclic, fused, spiro or bridged ring system which can be saturated or contain one or more units of unsaturation, having five to ten ring atoms in which one or more (e.g., one to three, or one, two, or three) ring atoms is a heteroatom selected from, N, S, and O. In some embodiments, the heterocycle comprises 5-6 ring members. In some embodiments, the heterocycle comprises 5 ring members. In some embodiments, the heterocycle comprises 6 ring members. Examples of heterocycles include, but are not limited to, quinuclidinyl, piperidinyl, piperizinyl, pyrolidinyl, pyrazolidinyl, imidazolidinyl, azepanyl, diazepanyl, triazepanyl, azocanyl, diazocanyl, triazocanyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, oxazocanyl, oxazepanyl, thiazepanyl, thiazocanyl, benzimidazolonyl, tetrahydrofuranyl, tetrahydrothiophenyl, morpholino (including, for example, 3-morpholino, 4-morpholino), 2-thiomorpholino, 3- thiomorpholino, 4-thiomorpholino, 1-pyrolidinyl, 2-pyrolidinyl, 3-pyrolidinyl, pyrolidin-2-one, 1- tetrahydropiperazinyl, 2-tetrahydropiperazinyl, 3-tetrahydropiperazinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 1- pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl, 5-pyrazolinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 2- thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl, 1-imidazolidinyl, 2-imidazolidinyl, 4-imidazolidinyl, 5-imidazolidinyl, indolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzothiolanyl, benzodithianyl, 3-(1-alkyl)-benzimidazol-2- onyl, and 1,3-dihydro-imidazol-2-onyl.

[0028] The term "aryl” refers to aromatic ring groups have only carbon ring atoms (i.e., six to ten carbon ring atoms) and include monocyclic aromatic rings such as phenyl and fused polycyclic aromatic ring systems in which two or more carbocyclic aromatic rings are fused to one another. In some embodiments, aryl is phenyl.

[0029] The terms "heteroaryl" refers to a heterocycle that is aromatic, having five to eight members (e.g., 5 to 6 members), including monocyclic heteroaromatic rings and polycyclic aromatic rings in which a monocyclic aromatic ring is fused to one or more other aromatic ring. Heteroaryl groups have one or more ring (e.g., 1 to 3, 1, 2, or 3,) heteroatoms selected from N, O, and S. Also included within the scope of the term "heteroaryl", as it is used herein, is a group in which an aromatic ring is "fused" to one or more non-aromatic rings (carbocyclic or heterocyclic), where the radical or point of atachment is on the aromatic ring. Examples of heteroaryl groups include pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, imidazolyl, pyrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl or thiadiazolyl including, for example, 2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5- imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxadiazolyl, 5-oxadiazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3- pyrazolyl, 4-pyrazolyl, 1-pyrolyl, 2-pyrolyl, 3-pyrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5- pyrimidinyl, 3-pyridazinyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-triazolyl, 5-triazolyl, tetrazolyl, 2-thienyl, 3-thienyl, , isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl, 1,3,4- thiadiazolyl, 1,2,5-thiadiazolyl, pyrazinyl, and 1,3,5-triazinyl. A heteroaryl ring is unsubstituted or substituted as described herein.

[0030] Compounds of the present disclosure can exist in particular geometric or stereoisomeric forms having one or more asymmetric carbon atoms. The present disclosure contemplates such forms, including cis- and trans- isomers, R- and S-enantiomers, diastereomers, racemic mixtures thereof, and other mixtures thereof, as faling within the scope of the disclosed compounds. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. Al such isomers, as wel as mixtures thereof, are intended for inclusion herein.

[0031] As used herein, the term “pharmaceuticaly acceptable” means that the referenced substance, such as a compound of the present disclosure, or a formulation containing the compound, or a particular excipient, are safe and suitable for administration to a patient or subject. The term “pharmaceuticaly acceptable excipient” refers to a medium that does not interfere with the efectiveness of the biological activity of the active ingredient(s) and is not toxic to the host to which it is administered.

[0032] Compositions may also include pharmaceuticaly acceptable salts of a compound disclosed herein (i.e., compounds of Formula (I), Compound 1, and compounds as disclosed in Table A).

[0033] The compounds disclosed herein can be as a pharmaceuticaly acceptable salt. As used herein, the term “pharmaceuticaly acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, iritation, alergic response and the like, and are commensurate with a reasonable benefit / risk ratio. Pharmaceuticaly acceptable salts are wel known in the art. For example, S. M. Berge et al. describe pharmaceuticaly acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, which is incorporated herein by reference. Pharmaceuticaly acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceuticaly acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, trifluoroacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Otherpharmaceuticaly acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, glutamate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Salts of compounds containing a carboxylic acid or other acidic functional group can be prepared by reacting with a suitable base. Such salts include, but are not limited to, alkali metal, alkaline earth metal, aluminum salts, ammonium, N+(C1- 4alkyl)4 salts, and salts of organic bases such as trimethylamine, triethylamine, morpholine, pyridine, piperidine, picoline, dicyclohexylamine, N,N'-dibenzylethylenediamine, 2-hydroxyethylamine, bis-(2-hydroxyethyl)amine, tri-(2- hydroxyethyl)amine, procaine, dibenzylpiperidine, dehydroabietylamine, N,N'-bisdehydroabietylamine, glucamine, N- methylglucamine, colidine, quinine, quinoline, and basic amino acids such as lysine and arginine. This invention also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water or oil-soluble or dispersible products may be obtained by such quaternization. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceuticaly acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate. Pharmaceutical Formulations, Dosing, and Routes of Administration

[0034] Further provided are pharmaceutical formulations (alternatively refered to as compositions throughout herein) comprising a compound as described herein or pharmaceuticaly acceptable salt thereof, and a pharmaceuticaly acceptable excipient.

[0035] The compositions administered to a subject may be conveniently formulated for administration with one or more pharmaceuticaly acceptable carier(s).

[0036] The compounds described herein can be administered to a subject in a therapeuticaly efective amount, alone or as part of a pharmaceuticaly acceptable composition or formulation. In addition, the compounds can be administered al at once, multiple times, or delivered substantialy uniformly over a period of time. It is also noted that the dose of the compound can be varied over time.

[0037] A particular administration regimen for a particular subject wil depend, in part, upon the compound, the amount of compound administered, the route of administration, and the cause and extent of any side efects. The amount of compound administered to a subject (e.g., a mammal, such as a human) in accordance with the disclosure should be suficient to afect the desired response over a reasonable time frame. Dosage typicaly depends upon the route, timing, and frequency of administration. Accordingly, the clinician titers the dosage and modifies the route of administration to obtain the optimal therapeutic efect, and conventional range-finding techniques are known to those of ordinary skil in the art.

[0038] Purely by way of ilustration, the method comprises administering, for example, from about 0.1 mg / kg up to about 100 mg / kg of compound or more, depending on the factors mentioned above. In other embodiments, the dosage ranges from 1 mg / kg up to about 100 mg / kg; or 5 mg / kg up to about 100 mg / kg; or 10 mg / kg up to about 100 mg / kg. Some conditions require prolonged treatment, which may or may not entail administering lower doses of compound over multiple administrations. If desired, a dose of the compound is administered as two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionaly, in unit dosage forms. The treatment period wil depend on the particular condition and symptoms, and may last one day to several months.

[0039] The compositions may be administered to a subject by any method. Methods of administration include but are not limited to parenteraly, subcutaneously, oraly, topicaly, pulmonarily, rectaly, vaginaly, intravenously, intraperitonealy, intrathecaly, intracerebraly, epiduraly, intramuscularly, intradermaly, or intracarotidly.

[0040] Suitable methods of administering a physiologicaly-acceptable composition, such as a pharmaceutical composition comprising the compounds disclosed herein are wel known in the art. Although more than one route can be used to administer a compound, a particular route can provide a more immediate and more efective reaction than another route. Depending on the circumstances, a pharmaceutical composition comprising the compound is applied or instiled into body cavities, absorbed through the skin or mucous membranes, ingested, inhaled, and / or introduced into circulation. For example, in certain circumstances, it wil be desirable to deliver a pharmaceutical composition comprising the agent oraly, through injection by intravenous, intraperitoneal, intracerebral (intra- parenchymal), intracerebroventricular, intramuscular, intra-ocular, intraarterial, intraportal, intralesional, intramedulary, intrathecal, intraventricular, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, urethral, vaginal, or rectal means, by sustained release systems, or by implantation devices. If desired, the compound is administered regionaly via intrathecal administration, intracerebral (intra-parenchymal) administration, intracerebroventricular administration, or intraarterial or intravenous administration feeding the region of interest. Alternatively, the composition is administered localy via implantation of a membrane, sponge, or another appropriate material onto which the desired compound has been absorbed or encapsulated. Where an implantation device is used, the device is, in one aspect, implanted into any suitable tissue or organ, and delivery of the desired compound is, for example, via difusion, timed-release bolus, or continuous administration.

[0041] To facilitate administration, the compound is, in various aspects, formulated into a physiologicaly- acceptable composition comprising a carier (e.g., vehicle, adjuvant, or diluent). The particular carier employed is limited only by physico-chemical considerations, such as solubility and lack of reactivity with the compound, and by the route of administration. Physiologicaly- acceptable cariers are wel known in the art. Ilustrative pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions (for example, see U.S. Patent No. 5,466,468). Injectable formulations are further described in, e.g., Pharmaceutics and Pharmacy Practice, J. B. Lippincot Co., Philadelphia. Pa., Banker and Chalmers, eds., pages 238-250 (1982), and ASHP Handbook on Injectable Drugs, Toissel, 4th ed., pages 622-630 (1986). A pharmaceutical composition comprising the compound is, in one aspect, placed within containers, along with packaging material that provides instructions regarding the useof such pharmaceutical compositions. Generaly, such instructions include a tangible expression describing the reagent concentration, as wel as, in certain embodiments, relative amounts of excipient ingredients or diluents (e.g., water, saline or PBS) that may be necessary to reconstitute the pharmaceutical composition.

[0042] Compositions suitable for parenteral injection may comprise physiologicaly acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Examples of suitable aqueous and nonaqueous cariers, diluents, solvents, or vehicles include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, and the like), suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0043] These compositions may also contain adjuvants such as preserving, weting, emulsifying, and dispersing agents. Microorganism contamination can be prevented by adding various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents, for example, sugars, sodium chloride, and the like. Prolonged absorption of injectable pharmaceutical compositions can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0044] Solid dosage forms for oral administration include capsules, tablets, powders, and granules. In such solid dosage forms, the active compound is admixed with at least one inert customary excipient (or carier) such as sodium citrate or dicalcium phosphate or (a) filers or extenders, as for example, starches, lactose, sucrose, mannitol, and silicic acid; (b) binders, as for example, carboxymethylcelulose, alginates, gelatin, polyvinylpyrolidone, sucrose, and acacia; (c) humectants, as for example, glycerol; (d) disintegrating agents, as for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (a) solution retarders, as for example, parafin; (f) absorption accelerators, as for example, quaternary ammonium compounds; (g) weting agents, as for example, cetyl alcohol and glycerol monostearate; (h) adsorbents, as for example, kaolin and bentonite; and (i) lubricants, as for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, and tablets, the dosage forms may also comprise bufering agents. Solid compositions of a similar type may also be used as filers in soft and hard filed gelatin capsules using such excipients as lactose or milk sugar, as wel as high molecular weight polyethylene glycols, and the like.

[0045] Solid dosage forms such as tablets, dragees, capsules, pils, and granules can be prepared with coatings and shels, such as enteric coatings and others wel known in the art. The solid dosage forms may also contain opacifying agents. Further, the solid dosage forms may be embedding compositions, such that they release the active compound or compounds in a certain part of the intestinal tract in a delayed manner. Examples of embedding compositions that can be used are polymeric substances and waxes. The active compound can also be in micro- encapsulated form, optionaly with one or more excipients.

[0046] Liquid dosage forms for oral administration include pharmaceuticaly acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, the liquid dosage form may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, as for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, in particular, cotonseed oil, groundnut oil, corn germ oil, olive oil, castor oil, and sesame seed oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and faty acid esters of sorbitan, or mixtures of these substances, and the like.

[0047] Besides such inert diluents, the composition can also include adjuvants, such as weting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. Suspensions, in addition to the active compound, may contain suspending agents, as for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystaline celulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, or mixtures of these substances, and the like.

[0048] Compositions for rectal administration are preferably suppositories, which can be prepared by mixing the compounds of the disclosure with suitable non-iritating excipients or cariers such as cocoa buter, polyethylene glycol or a suppository wax, which are solid at ordinary room temperature, but liquid at body temperature, and therefore, melt in the rectum or vaginal cavity and release the active component.

[0049] The compositions used in the methods of the disclosure may be formulated in miceles or liposomes. Such formulations include stericaly stabilized miceles or liposomes and stericaly stabilized mixed miceles or liposomes. Such formulations can facilitate intracelular delivery, since lipid bilayers of liposomes and miceles are known to fuse with the plasma membrane of cels and deliver entrapped contents into the intracelular compartment.

[0050] Upon formulation, solutions wil be administered in a manner compatible with the dosage formulation and in such amount as is therapeuticaly efective. The formulations are easily administered in a variety of dosage forms such as injectable solutions, drug release capsules and the like. For parenteral administration in an aqueous solution, for example, the solution should be suitably bufered if necessary and the liquid diluent first rendered isotonic with suficient saline or glucose. These particular aqueous solutions are especialy suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration.

[0051] The frequency of dosing wil depend on the pharmacokinetic parameters of the agents and the routes of administration. The optimal pharmaceutical formulation wil be determined by one of skil in the art depending on the route of administration and the desired dosage. See, for example, Remington’s Pharmaceutical Sciences, 18th Ed. (1990) Mack Publishing Co., Easton, PA, pages 1435-1712, incorporated herein by reference. Such formulations may influence the physical state, stability, rate of in vivo release and rate of in vivo clearance of the administered agents. Depending on the route of administration, a suitable dose may be calculated according to body weight, body surface areas or organ size. Further refinement of the calculations necessary to determine the appropriate treatment dose is routinely made by those of ordinary skil in the art without undue experimentation, especialy in light of the dosage information and assays disclosed herein, as wel as the pharmacokinetic data observed in animals or human clinical trials.

[0052] The precise dosage to be employed depends upon several factors including the host, whether in veterinary medicine or human medicine, the nature and severity of the condition, e.g., disease or disorder, being treated, the mode of administration and the particular active substance employed. The compounds may be administered by any conventional route, in particular enteraly, and, in one aspect, oraly in the form of tablets or capsules. Administered compounds can be in the free form or pharmaceuticaly acceptable salt form as appropriate, for use as a pharmaceutical, particularly for use in the prophylactic or curative treatment of a disease of interest. These measures wil slow the rate of progress of the disease state and assist the body in reversing the process direction in a natural manner.

[0053] It wil be appreciated that the pharmaceutical compositions and treatment methods of the invention are useful in fields of human medicine and veterinary medicine. Thus, the subject to be treated is in one aspect a mammal. In another aspect, the mammal is a human.

[0054] In jurisdictions that forbid the patenting of methods that are practiced on the human body, the meaning of “administering” of a composition to a human subject shal be restricted to prescribing a controled substance that a human subject wil self-administer by any technique (e.g., oraly, inhalation, topical application, injection, insertion, etc.). The broadest reasonable interpretation that is consistent with laws or regulations defining patentable subject mater is intended. In jurisdictions that do not forbid the patenting of methods that are practiced on the human body, the “administering” of compositions includes both methods practiced on the human body and also the foregoing activities.

[0055] In another embodiment, it is envisioned that the compositions are used in combination with other drugs including but not limited to additional neurological disease therapeutics, anti-cancer drugs, anti-inflammatory drugs, and / or immune-modulatory drugs. In some cases, the one or more additional therapeutics comprise a neurological disease therapeutic, an anti-cancer drug, an anti-inflammatory drug, an immune-modulatory drug, and combinations thereof. Methods of Synthesizing the Compounds of the Disclosure

[0056] Provided herein are processes of synthesizing a compound of Formula (I), or a salt thereof:R’ is OR1’, SR1’, NRa’Rb’, S(O)R1’, S(O2)R1’, or P(O2)R1’; R1’ is C1-6alkyl, C2-6alkenyl, C1-6alkylene-5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S, C3-10cycloalkyl, C6-10aryl, 5- to 10 membered heterocycloalkyl comprising one to three heteroatoms selected from O, N, and S, or 5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N,and S; and Ra’ and Rb’ are individualy H, C1-6alkyl, C2-6alkenyl, C(O)OC1-6alkyl, C3-10cycloalkyl, C6-10aryl, 5- to 10 membered heterocycloalkyl comprising one to three heteroatoms selected from O, N, and S, or 5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S, or C1-6alkylene-5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S; the method comprising a cyclization step comprising admixing a compound of Formula (a) with (Z)-1-cyclopropyl-3- (dimethylamino)-2-(3-methylisoxazol-5-yl)prop-2-en-1-one (“Compound 12”) in the presence of a base and a polar solvent: , reaction between the compound of Formula (a) and Compound 12.

[0057] In some cases, R’ is C1-6alkylene-5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S. In some cases, R’ is C2alkylene-5-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S. In some cases, the compound of . In some cases, the compound of Formula (I) has the structure of Compound.is OR1’, SR1’, or NRa’Rb’. In some cases, R’ is OR1’ or SR1’. In some cases, R’ is OR1’. In some cases, R’ is SR1’. In some cases, R1’ is C1-6alkyl, C2-6alkenyl, C3-10cycloalkyl, C6-10aryl, 5- to 10 membered heterocycloalkyl comprising one to three heteroatoms selected from O, N, and S, or 5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S. In some cases, R1’ is C1-6alkyl. In some cases, R1’ is methyl. In some cases, R’ is NRa’Rb’. In some cases, Ra’ is H. In some cases, Rb’ is H. In some cases, Rb’ is C1- 6alkyl or C(O)OC1-6alkyl. In some cases, Rb’ is C1-6alkyl. In some cases, Rb’ is methyl. In some cases, Rb’ is C(O)OC1-6alkyl. In some cases, Rb’ is C(O)O-t-butyl. In some cases, R’ is SMe, OMe, NH2, NHMe, or NHBoc. In some cases, R’ is OMe or SMe. In some cases, R’ is OMe. In some cases, R’ is SMe. In some cases, R’ is NH2, NHMe, or NHBoc. In some cases, R’ is NH2. In some cases, R’ is NHMe. In some cases, R’ is NHBoc.

[0059] In some cases, the compound of Formula (I) or salt thereof is a compound of Formula (I) or a salt thereof, i.e., provided are processes of synthesizing a compound of Formula (I), or a salt thereof.

[0060] In some cases, prior to the cyclization step the process comprises: (i) a condensation step comprising admixing 3,5-dimethylisoxazole and methyl cyclopropane carboxylate in the presence of a base in a polar solvent, under conditions suficient to efect a condensation reaction to yield 1- cyclopropyl-2-(3-methylisoxazol-5-yl)ethan-1-one (“Compound 11”): admixing Compound 11 and N,N-dimethylformamide dimethyl acetal (DMF-to form Compound 12. In some cases, the condensation step (i) comprises admixing 3,5-dimethylisoxazole and methyl cyclopropane carboxylate in the presence of a base in a polar aprotic solvent. In some cases, the polar aprotic solvent is tetrahydrofuran (THF). In some cases, the base is lithium disopropylamide (LDA).

[0061] In some cases, the admixing is caried out at a temperature of - 70 °C or lower. In some cases, the admixing is caried out at a temperature of about - 78 °C. In some cases, the condensation step (i) comprises the steps of: (ia) combining 3,5-dimethylisoxazole and THF to form a first solution; (ib) admixing the first solution with LDA that has been pre-cooled to a temperature of - 70 °C or lower to form a second solution; and (ic) admixing the second solution with methyl cyclopropane carboxylate under conditions suficient to yield Compound 11. In some cases, the methyl cyclopropane carboxylate is provided as a solution of methyl cyclopropane carboxylate in THF. In some cases, the LDA has been pre-cooled to a temperature of about - 78 °C. Methods of Use

[0062] Certain compositions may be used for the treatment of disease. In one embodiment, the diseases may include but are not limited to neurological disorders and diseases, cancer, and other like diseases and disorders. In one embodiment, neurological disorders and diseases include but are not limited to Alzheimer’s disease, Parkinson’s disease, neurodevelopmental disorders, traumatic brain injury, stroke, Amyotrophic Lateral Sclerosis, Huntington’s disease, ischemia, atention deficit disorders, and epilepsy. In one embodiment, cancer includes breast cancer, liver cancer, prostate cancer, leukemia (ex. Acute lymphoblastic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia), lymphoma (ex. Hodgkin lymphoma, Non-Hodgkin lymphoma), multiple myeloma, pancreatic cancer, colon cancer, thyroid cancer, bladder cancer, neuroblastoma, brain cancers (gliomas, meningiomas, pituitary adenomas etc.), lung cancer, ovarian cancer, stomach cancer, skin cancer (melanoma), cervical cancer, testicular cancer, kidney cancer, carcinoid tumors, and bone cancer. In one embodiment, the disease is associated with aberant 17β-HSD10 activity.

[0063] Thus, provided herein are methods of modulating 17β-HSD10 activity, comprising contacting a cel with a therapeuticaly efective amount of the compound or salt disclosed herein (i.e., a compound of Formula (I), Compound 1, or a compound disclosed in Table A, or a pharmaceuticaly acceptable salt thereof). Further provided are methods of modulating 17β-HSD10 activity in a subject, comprising administering to the subject a therapeuticaly efective amount of a compound or salt disclosed herein (i.e., a compound of Formula (I), Compound 1, or a compound disclosed in Table A, or a pharmaceuticaly acceptable salt thereof).

[0064] Also provided are methods of treating, inhibiting, or preventing a disease or disorder in a subject in need thereof, comprising administering to a subject a therapeuticaly efective amount of a compound or salt disclosed herein (i.e., a compound of Formula (I), Compound 1, or a compound disclosed in Table A, or a pharmaceuticaly acceptable salt thereof). In some cases, the disease or disorder is associated with aberant 17β-HSD10 activity. In some cases, the disease or disorder is a neurological disease or disorder, cancer, and other like diseases and disorders. In some cases, the neurological disorder or disease is Alzheimer’s disease, Parkinson’s disease, a neurodevelopmental disorder, traumatic brain injury, stroke, Amyotrophic Lateral Sclerosis, Huntington’s disease, ischemia, atention deficit disorders, or epilepsy. In some cases, the neurological disorder or disease is Alzheimer’s disease. In some cases, the disease or disorder is a neurological disease or disorder. In some cases, the disease or disorder is cancer.

[0065] Specificaly contemplated are methods of using a therapeuticaly efective amount of a compound disclosed herein for use as a therapeutic in a subject. As used herein, the term “therapeuticaly efective amount” means an amount of a compound or combination of therapeuticaly active compounds (i.e., a compound of Formula (I), Compound 1, or a compound disclosed in Table A, or a pharmaceuticaly acceptable salt thereof) that ameliorates, atenuates or eliminates one or more symptoms of a particular disease or condition (e.g., a neurological disease or disorder such as Alzheimer’s disease, or cancer), or prevents or delays the onset of one of more symptoms of a particular disease or condition.

[0066] The therapeuticaly efective amount can vary depending upon the intended application, or the subject and disease condition being treated, e.g., the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the weight and age of the patient, which can readily be determined by one of ordinary skil in the art. The term also applies to a dose that wil induce a particular response in target cels, e.g., modulating or inhibiting 17β-HSD10 activity in a cel to prevent or treat a neurological disorder, for example Alzheimer’s disease). The specific dose wil vary depending on, for example, the particular compounds chosen, the species of subject and their age / existing health conditions or risk for health conditions, the dosing regimen to be folowed, the severity of the disease, whether it is administered in combination with other agents, timing of administration, the tissue to which it is administered, and the physical delivery system in which it is caried.

[0067] As used herein, the term “subject” refers to animals, such as dogs, cats, cows, horses, and sheep (e.g., non-human animals) and humans. Particular subjects are mammals (e.g., humans). The term “subject” includes males and females.

[0068] As used herein, the terms "treatment" or "treating" a disease or disorder refers to a method of reducing, delaying or ameliorating such a condition before or after it has occured. The terms "prevention" or "preventing" a disease or disorder specificaly refers to a method of reducing, delaying or ameliorating such a condition before it has occured. Treatment may be directed at one or more efects or symptoms of a disease and / or the underlying pathology. Treatment is aimed to obtain beneficial or desired results including, but not limited to, therapeutic benefit and / or a prophylactic benefit. By “therapeutic benefit” is meant eradication or amelioration of the underlying disorder being treated. Also, a therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the patient, notwithstanding that the patient can stil be aflicted with the underlying disorder. For prophylactic benefit, the pharmaceutical compounds and / or compositions can be administered to a patient at risk of developing a particular disease, or to a patient reporting one or more of the physiological symptoms of a disease, even though a diagnosis of this disease may not have been made. The treatment can be any reduction and can be, but is not limited to, the complete ablation of the disease or the symptoms of the disease. As compared with an equivalent untreated control, such reduction or degree of prevention is at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, or 100% as measured by any standard technique.

[0069] As used herein, the term "therapeutic efect" refers to a therapeutic benefit and / or a prophylactic benefit as described herein. A prophylactic efect includes delaying or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof. EXAMPLES Example 1: Synthesis of BCC0100281 Intermediates

[0070] The screening library constituent BCC0100281 (Compound 1) was one of 16 smal molecules identified as inhibitors of 17β-HSD10 in a high-throughput screen of 6759 compounds.37 Due to the favorable physiochemical properties and moderate inhibitory activity (pIC504.6), compound 1 was identified as a compound of interest with ‘drug-like’ potential. To confirm activity of 1 and access analogues for medicinal chemistry optimization studies, synthetic access was required. Surprisingly, no synthetic route to access 1 or similar analogues has been disclosed in the literature. Thus, provided herein are the total synthesis of BCC0100281 (1) suitable for analogue generation, in silico and biochemical binding studies for confirming 17β-HSD10 target engagement, and biological activity of selected intermediate derivatives as chemical probes.

[0071] Retrosynthetic analysis (Scheme 1) identified that 1 could be obtained through two key intermediates, a suitably functionalized guanidine and an α,β unsaturated ketone. These intermediates can be obtained from commercialy available building blocks in two linear sequences which converge finaly to aford 1 through a condensation reaction.

[0072] To synthesize the first required intermediate, guanidine 8 was obtained in six steps from alcohol 2 (Scheme 2). Alcohol 2 can be accessed in two steps from 2,3-dihydrofuran38, 39 and is commercialy available. Chlorination of 2 was accomplished in quantitative yield upon exposure to thionyl chloride to aford 3 which was subsequently treated with biphasic conditions to facilitate tert-butyloxycarbonyl (Boc) protection of the pyrazolenitrogen resulting in chloride 4. Chloride 4 proved an efective electrophilic substrate for the alkylation of methylbenzylamine to aford tertiary amine 5b with a 11% yield. The low yield of 5b was atributed to a complex mixture of side products obtained from the reaction (Table B), with Boc deprotection occuring in situ to aford both 3 and 5a alongside elimination product 9. Reaction condition screening revealed that acetonitrile was required as solvent, and a temperature of 90 °C most efectively suppressed formation of the elimination product 9 (7%) while yielding 5a, removing the need for the Boc deprotection step. Furthermore, chloride 3 could be isolated upon purification and progressed back through the synthetic scheme, while Boc product 5b could undergo deprotection to give 5a, afording an overal yield based on recovered starting material of 71%.Entry Base Solvent Temp. °C 4 5a 5b 3 9 1 NaH THF rt aNo Reaction 2 K2CO4 DMF 70 bNo Reaction 3 NaHCO3 DMF 70 bNo Reaction c4 NaHCO3 MeCN 70 43 11 34 9 3 c5 NaHCO3 MeCN 90 6 54 12 21 7 c6 NaHCO3 MeCN 110 0 55 0 15 30 Table B. Reaction trials for the synthesis of 5a / b. Reaction conditions: To 4 (0.87 mmol, 1.0 equiv) in solvent (2.0 mL) was added N-methyl-1-phenylmethanamine (1.30 mmol, 1.5 equiv) and base (2.60 mmol, 3 equiv) aReaction analysis (TLC / HPLC) showed only starting material. bReaction analysis showed no starting material remaining, NMR of isolated material showed no evidence of compounds 3, 4, 5a, 5b, 9, 10. cRatio quantification of 5a, 5b, 3, and 9 achieved by HPLC.

[0073] Due to the Boc deprotection occuring in situ, the reaction was atempted with the unprotected pyrazole 3 but this proved unsuccessful. Alternative pyrazole-protecting groups (Table C) were introduced to improve ease of purification. The benzyl, p-methoxybenzyl, and tosyl protecting groups were selected given their increased stability to withstand the harsher reaction conditions, including high temperatures. The synthesis of 5c-e was achieved folowing a longer route through reaction with potassium phthalimide folowed by deprotection, reductive amination, and methylation. This method proved successful in obtaining 5c-e with no premature elimination or deprotection. However, this strategy proved futile as deprotection of al three pyrazole-protecting group under multiple conditions was unsuccessful (Table C). Treating the benzyl (5c) and p-methoxybenzyl (5d) pyrazole compounds with hydrogen and paladium on carbon aforded only deprotection of the aliphatic amine (to aford 6c and 6d respectively), with pyrazole deprotection not occuring at temperatures up to 90 °C and pressure up to 7 bar (entries 1 – 5). Alternative conditions for the PMB deprotection (entries 6-9) were atempted, and although some conditions (entries 7 & 8) showed evidence of product by MS, isolation was unsuccessful. Deprotection of tosyl pyrazole (5e) also proved unsuccessful under both acidic and basic conditions (entries 10 & 11).Entry Substrate Conditions Product 1 5c H2 (1 bar), Pd / C (20%), MeOH (2 mL), rt 6c 2 5c H2 (1 bar), Pd / C (20%), MeOH (2 mL), 90 °C 6c 3 5c H2 (7 bar), Pd / C (20%), MeOH (2 mL), rt 6c 4 5c H2 (7 bar), Pd / C (20%), MeOH (2 mL), 90 °C 6c 5 5d H2 (7 bar), Pd / C (20%), MeOH (2 mL), 90 °C 6d 6 5d TFA (10 equiv), MeOH (2 mL), 70 °C 6d 7 5d TFA (47 equiv), 100 °C a6a 8 5d DDQ (1.2 equiv), DCM:H2O (20:4) (5.25 mL), rt a6a 9 5d Ceric ammonium nitrate (4 equiv), MeCN:H2O (3:1) (4 mL), rt 6d 10b 5e AcOH:H2SO4 (1:1) (5 mL) no product 11b 5e NaOH (4M):MeOH:THF 2:1:4 (3.5 mL) 80 °C no product Table C. Alternative protecting groups and deprotection conditions. aProduct detected by HRMS however no product obtained from purification. b No starting material remaining after 4h, no product mass or peak observed.

[0074] Hydrogenation of benzylamine 5a using paladium on carbon aforded secondary amine 6a in quantitative yield. Reaction of 6a with bis-Boc thiourea in the presence of mercury (I) chloride and triethylamine aforded Bis- Boc-protected guanidine 7 which subsequently underwent Boc deprotection when exposed to trifluoroacetic acid, as described by Kim et al.40 to aford desired intermediate 8.

[0075] The second Intermediate, α,β unsaturated ketone 12, required for the synthesis of 1, was obtained in two steps from commercialy available isoxazole 10 (Scheme 3). First described by Brunele,41 the lithiation of 10 with lithium disopropylamide (LDA) at low temperature (-78 °C) occurs regiospecificaly at the C5-methyl group. Exploiting this regioselectivity and folowing a similar procedure reported by Del Belo et al,42 intermediate 11 was obtained in 51% yield initialy. However, reversing the addition of reagents, i.e. adding a solution of 10 in THF to pre- cooled LDA, improved this yield to 92%. The subsequent reaction of ketone 11 and N,N-dimethylformamide dimethyl acetal (DMF-DMA) occurs through refluxing in toluene overnight to aford desired intermediate 12.

[0076] The final step in the synthesis of 1 combines intermediates 8 and 12 (Scheme 4) in the presence of potassium carbonate to form the 2,4,5-substituted pyrimidine ring of 1. Example 2: Synthesis of BCC0100281 Analogs

[0077] The cyclization step described in Scheme 4 for the synthesis of 1 was optimized using a model reaction composed of commercialy available guanidine and urea-like building blocks (Table D) prior to atempt with 8. Alteration of the base and solvent from sodium ethoxide and ethanol to their non-nucleophilic counterparts potassium carbonate and dioxane (additionaly acetonitrile can also be used as the solvent) improved overal the yield of this cyclization to 90% after purification, and in some cases, no purification was required. This process provided access to analogs of 1 (Compounds 14a-14e shown in Table 3).Yield 1 a13a NaOMe MeOH 65 °C 14a 32% 2 13b K2CO3 dioxane 100 °C 14b 11% 3 13c K2CO3 dioxane 100 °C 14c b100% 4 13d K2CO3 MeCN 100 °C 14d 72% 5 13e K2CO3 dioxane 100 °C 14e 17% 6 13e K2CO3 MeCN 100 °C 14e 22% Table D. Optimization of conditions and substrates for the cyclization of guanidine and urea building blocks with 12. Reaction conditions: 1 (1 equiv), 13a-e (1.2 equiv), base (3 equiv), solvent (0.1-0.3M). a13a (2.4 equiv), base (3.4 equiv) bNo purification of product required. Example 3: in silico Binding Studies of 1 to 17β-HSD10

[0078] With a synthetic route suitable for access to hit compound 1 and generation of derivatives in hand, the ability of 1 to bind to the 17β-HSD10 protein target was determined. In silico binding studies of 1 were conductedusing the published crystal structure of the NADPH 17β-HSD10 homo tetramer in complex with the inhibitor AG18051 (PDBID: 1U7T).43 Employing SeeSAR 12.1 software (BioSolveIT), the AG18051-NADPH ligand was removed and the binding site of 61 amino acids defined. Compound 1 was prepared and docked using HYDE to determine predicted binding interactions (FIG.1). The pyrazole ring was predicted to form hydrogen bonds with amino acid residues Valine 65 and Alanine 63, the pyrimidine nitrogen distal to the cyclopropyl ring was predicted to from a hydrogen bond with Glycine 93. The oxygen of the isoxazole ring was predicted to form a hydrogen bond with amino acid residue Glycine 23 and the nitrogen with Leucine 22. Thus, in silco modeling supports compound 1 binding 17β-HSD10. Example 4: in vitro Binding Studies of 1 to 17β-HSD10

[0079] To confirm binding of 1 to 17β-HSD10, a protein thermal shift assay was conducted. Upon titration of 1 to 17β-HSD10, the melting temperature of the protein decreased. This change in melting temperature was concentration-dependent with increasing concentration of 1 ranging from 42.29 °C with 5 mM treatment of 1, a change of -0.37 °C from the native protein, to 40.46 °C with 100 µM treatment of 1, a change of -2.2 °C from the native protein (FIG.2). Solubility of 1 hindered testing at higher concentrations. The reduction of melting temperature suggested that 1-17β-HSD10 complex formation decreased the thermal stability of the protein and confirmed binding. This is consistent with published studies where the structuraly-dynamic loops forming part of the 17β- HSD10 active site undergoes a conformational change upon substrate binding.43-45 These data suggest that binding of compound 1 to 17β-HSD10 may stimulate structural changes resulting in thermal destabilization and that the conformation of 17β-HSD10 in complex with 1 is less stable than the apo-protein conformation. Example 5: Cel Viability Studies

[0080] With high confidence Compound 1 binds 17β-HSD10, the toxicity of compound 1 in three cel lines known to overexpress 17β-HSD10 was assessed.46, 47 The SH-SY5Y neuroblastoma cel line was employed due to exhibiting neurite structure and expressing immature neuronal markers, thus serving as a cel model for AD studies. Both LNCaP and PANC-1, prostate and pancreatic cancer cel lines respectively were used to assess the toxicity of 1 in cancer cels (FIGs.3A, 3B, and 3C). The compound was found to exhibit toxicity in SH-SY5Y cels at concentrations greater than 10 mM (FIG.3A). The compound shows specific toxicity towards neuroblastoma cel lines SK-N-MC, SK-N-AS, CHLA255 and the MYCN amplified cel lines SK-N-BE2 at concentrations greater than 80 µM. However, in LNCap (FIG.3B) and PANC-1 cels (FIG.3C), 1 exhibited no toxicity at concentrations up to 100 mM. Example 6: Amelioration of Aβ Toxicity with 1 and BCC0100281 Analogs

[0081] Modulators of 17β-HSD10 have the potential to disrupt binding of Aβ and, therefore, provide protective efects against Aβ toxicity. The ability of 1 to ameliorate Aβ1-42 toxicity in SH-SY5Y cels was therefore assessed. The co-treatment of cels with Aβ and 1 showed that this compound provides no amelioration of Aβ1-42 toxic efects. (FIGs.4A and 4B).

[0082] While compound 1 provides no protective efects to ameliorate Aβ-induced toxicity, the patern of cel viability closely matches the toxicity patern seen with the compound in SH-SY5Y cels. Without wishing to be boundby any particular theory, protection of Aβ toxicity may be masked by this innate toxic efect. Gratifyingly, the compound is not indiscriminately toxic, with litle efect seen in LNCaP and PANC-1 cels (FIGs.4A and 4B). The potential of selected synthetic derivatives to ameliorate Aβ-induced toxicity was therefore studied.

[0083] Compounds 14b-e represent a discrete series for determination of an initial structure-activity relationship (SAR) (FIGs.5A, 5B, 5C, and 5D), difering by para-substituent of the central phenyl ring. Methoxy compound 14b and Boc protected amine 14e confered no efect to ameliorate Aβ-induced toxicity (FIGs.5A and 5D). However, aniline 14c aforded significant protection of Aβ-induced toxicity at 0.1 µM concentration (FIG.5B). Additionaly, N- methylation to yield 14d aforded significant protection at 1 µM with a trend of protection at 0.1 µM; however, at concentrations above 50 µM, toxicity was observed. Without wishing to be bound by any particular theory, replacement with a methoxy moiety and Boc protection removes or reduces the ability of the amine hydrogen to participate in hydrogen bond donating, while adding an electron-donating group improves potential to form a stronger hydrogen bond by donation. This smal initial SAR study suggests the need for a para-basic nitrogen in the molecule and suggests strategies for further optimization.

[0084] In conclusion, provided herein is the first synthetic route to access screening library compound BCC0100281 (1) in nine steps with the longest linear sequence of seven with an overal yield of 0.7%. The route is suitable for the generation of intermediates and parent compound derivatives for SAR exploration. Furthermore, preliminary biological assessments to confirm the interaction of Compound 1 with 17β-HSD10 have been undertaken. Compound 1 was shown to be toxic to SH-SY5Y cels, precluding it from further studies in this cel line. However, intermediates containing the isoxazole ring show significant protection of cel viability against Aβ-induced toxicity. A discrete series revealed an initial SAR requirement for a hydrogen bond donating capable amine at the para-position of the phenyl ring, revealing a novel chemotype for further exploration as in Alzheimer’s therapeutic development. REFERENCES 1. X.-Y. He et al., Journal of Biological Chemistry, 1999, 274, 15014-15019. 2. X.-Y. He, et al., Journal of Biological Chemistry, 1998, 273, 10741-10746. 3. J. Zschocke, J Inherit Metab Dis, 2012, 35, 81-89. 4. J. Holzmann et al., Cel, 2008, 135, 462-474. 5. A. Morsy et al., Journal of Medicinal Chemistry, 2019, 62, 4252-4264. 6. S. Y. Yang et al., Trends Endocrinol Metab, 2005, 16, 167-175. 7. M. P. Murphy et al., J Alzheimers Dis, 2010, 19, 311-323. 8. X. Chen et al., IUBMB Life, 2006, 58, 686-694. 9. J. W. Lustbader et al., Science, 2004, 304, 448-452. 10. A. Jekabsone et al., International Journal of Molecular Sciences, 2023, 24, 12315. 11. A. Grimm et al., Mol Neurobiol, 2012, 46, 151-160. 12. S. Du Yan et al., Journal of Biological Chemistry, 1999, 274, 2145-2156. 13. A. T. Marques et al., Mini Rev Med Chem, 2009, 9, 1002-1008. 14. T. K. Man et al., Cancer Res, 2005, 65, 8142-8150. 15. S. Goodenough et al., Neuroscience, 2005, 132, 581-589. 16. J. Yao et al., Neurobiol Aging, 2012, 33, 1507-1521. 17. E. Vegeto et al., Endocrinology, 2006, 147, 2263-2272. 18. E. Vegeto et al., Experimental Gerontology, 2000, 35, 1309-1316. 19. D. L. Lei et al., Neuroscience, 2003, 121, 659-666. 20. M. Alvarez-de-la-Rosa et al., Ann N Y Acad Sci, 2005, 1052, 210-224.21. A. Vila et al., Endocr Rev, 2016, 37, 372-402. 22. J. Yao et al., Mol Cel Neurosci, 2007, 35, 377-382. 23. J.-J. Pei et al., Brain Research, 1998, 797, 267-277. 24. O. Kjaerulf et al., Cel Biochemistry and Biophysics, 2011, 60, 137-154. 25. Y. Ren et al., J Biol Chem, 2008, 283, 5685-5691. 26. S. Mehan et al., J Mol Neurosci, 2011, 43, 376-390. 27. M. Hanzlova et al., ACS Med Chem Let, 2023, 14, 1724-1732. 28. V. Metodieva et al., eNeuro, 2022, 9. 29. L. Aitken et al., Molecules, 2019, 24. 30. A. Morsy et al., ACS Chemical Neuroscience, 2022, 13, 2176-2190. 31. G. Aurilio et al., Cels, 2020, 9. 32. D. Ayan, et al., ChemMedChem, 2012, 7, 1181-1184. 33. X.-Y. He et al., The Journal of Steroid Biochemistry and Molecular Biology, 2003, 87, 191-198. 34. S. Salas et al., International Journal of Cancer, 2009, 125, 851-860. 35. O. Benek et al., Medicinal Chemistry, 2017, 13, 345-358. 36. E. A. Carlson et al., BMC Cancer, 2015, 15, 166. 37. L. Aitken et al., SLAS DISCOVERY: Advancing the Science of Drug Discovery, 2017, 22, 676-685. 38. S. P. Ivonin et al., Chemistry of Heterocyclic Compounds, 2020, 56, 320-325. 39. R. G. Jones et al., Journal of the American Chemical Society, 1953, 75, 4048-4052. 40. K. S. Kim et al., Tetrahedron Leters, 1993, 34, 7677-7680. 41. D. J. Brunele, Tetrahedron Leters, 1981, 22, 3699-3702. 42. F. Del Belo et al., Bioorganic & Medicinal Chemistry, 2015, 23, 5725-5733. 43. C. R. Kissinger et al., Journal of molecular biology, 2004, 342, 943-952. 44. Y. Yan et al., Biochemistry, 2007, 46, 1724-1731. 45. A. Powel et al., Journal of molecular biology, 2000, 303, 311-327. 46. M. Uhlén et al., Science, 2015, 347, 1260419. 47. T. H. P. Atlas, HSD17B10 - Cel Line, htps: / www.proteinatlas.org / ENSG00000072506- HSD17B10 / cel+line, (accessed 1-November, 2023).

Claims

What is Claimed:

1. A compound, or a pharmaceuticaly acceptable salt thereof, having the structure of Formula (I):R is OR1, SR1, NRaRb, S(O)R1, S(O2)R1, or P(O2)R1; R1 is C1-6alkyl, C2-6alkenyl, C3-10cycloalkyl, C6-10aryl, 5- to 10 membered heterocycloalkyl comprising one to three heteroatoms selected from O, N, and S, or 5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S; and Ra and Rb are individualy H, C1-6alkyl, C2-6alkenyl, C(O)OC1-6alkyl, C3-10cycloalkyl, C6-10aryl, 5- to 103. The compound or salt of claim 1 or 2, wherein R is OR1 or SR1.

4. The compound or salt of any one of claims 1 to 3, wherein R1 is C1-6alkyl.

5. The compound or salt of any one of claims 1 to 4, wherein R1 is methyl.

6. The compound or salt of claim 1 or 2, wherein R is NRaRb.

7. The compound or salt of claim 6, wherein Ra is H.

8. The compound or salt of claim 6 or 7, wherein Rb is H.

9. The compound or salt of claim 6 or 7, wherein Rb is C1-6alkyl or C(O)OC1-6alkyl.

10. The compound or salt of claim 9, wherein Rb is C1-6alkyl.

11. The compound or salt of claim 10, wherein Rb is methyl.

12. The compound or salt of claim 9, wherein Rb is C(O)OC1-6alkyl.

13. The compound or salt of claim 12, wherein Rb is C(O)O-t-butyl.

14. A compound, as recited in Table 1, or a pharmaceuticaly acceptable salt thereof.

15. The compound of claim 14 which is Compound 14c, 14d, or a pharmaceuticaly acceptable salt thereof.

16. The compound of any one of claims 1 to 15, in the form of a pharmaceuticaly acceptable salt.

17. A pharmaceutical composition comprising the compound or salt of any one of claims 1 to 16 and a pharmaceuticaly acceptable carier or excipient.

18. The pharmaceutical composition of claim 17, further comprising one or more additional therapeutics.

19. The pharmaceutical composition of claim 18, wherein the one or more additional therapeutics comprise a neurological disease therapeutic, an anti-cancer drug, an anti-inflammatory drug, an immune-modulatory drug, or a combination thereof.

20. A method of modulating 17β-HSD10 activity, comprising contacting a cel with a therapeuticaly efective amount of the compound or salt of any one of claims 1 to 16 or the pharmaceutical composition of claim 17.

21. A method of modulating 17β-HSD10 activity in a subject, comprising administering to the subject a therapeuticaly efective amount of the compound or salt of any one of claims 1 to 16 or the pharmaceutical composition of claim 17.

22. A method of treating, inhibiting, or preventing a disease or disorder in a subject in need thereof, comprising administering to a subject a therapeuticaly efective amount of the compound or salt of any one of claims 1 to 16 or the pharmaceutical composition of claim 17.

23. The method of claim 22, wherein the disease or disorder is associated with aberant 17β-HSD10 activity.

24. The method of claim 22 or 23, wherein the disease or disorder is a neurological disease or disorder, or cancer.

25. The method of claim 24, wherein the neurological disorder or disease is Alzheimer’s disease, Parkinson’s disease, a neurodevelopmental disorder, traumatic brain injury, stroke, Amyotrophic Lateral Sclerosis, Huntington’s disease, ischemia, atention deficit disorders, or epilepsy.

26. The method of claim 24 or 25, wherein the neurological disorder or disease is Alzheimer’s disease.

27. The method of any one of claims 21 to 26, further comprising administering one or more additional therapeutics to the subject.

28. The method of claim 27, wherein the one or more additional therapeutics comprise a neurological disease therapeutic, an anti-cancer drug, an anti-inflammatory drug, an immune-modulatory drug, or a combination thereof.

29. The compound or salt of any one of claims 1 to 16 or the pharmaceutical composition of claim 17, for use in modulating 17β-HSD10 activity.

30. The compound or salt of any one of claims 1 to 16 or the pharmaceutical composition of claim 17, for use in modulating 17β-HSD10 activity in a subject.

31. The compound or salt of any one of claims 1 to 16 or the pharmaceutical composition of claim 17, for use in treating, inhibiting, or preventing a disease or disorder in a subject in need thereof.

32. The compound, salt, or pharmaceutical composition for use of claim 31, wherein the disease or disorder is associated with aberant 17β-HSD10 activity.

33. The compound, salt, or pharmaceutical composition for use of claim 31 or 32, wherein the disease or disorder is a neurological disease or disorder, or cancer.

34. The compound, salt, or pharmaceutical composition for use of claim 33, wherein the neurological disorder or disease is Alzheimer’s disease, Parkinson’s disease, a neurodevelopmental disorder, traumatic brain injury, stroke, Amyotrophic Lateral Sclerosis, Huntington’s disease, ischemia, atention deficit disorders, or epilepsy.

35. The compound, salt, or pharmaceutical composition for use of claim 33 or 34, wherein the neurological disorder or disease is Alzheimer’s disease.

36. The compound, salt, or pharmaceutical composition for use of any one of claims 30 to 35, further formulated for use with one or more additional therapeutics.

37. The compound or composition for use of claim 36, wherein the one or more additional therapeutics comprise a neurological disease therapeutic, an anti-cancer drug, an anti-inflammatory drug, an immune-modulatory drug, or a combination thereof.

38. A process of synthesizing a compound of Formula (I), or a salt thereof:R’ is OR1’, SR1’, NRa’Rb’, S(O)R1’, S(O2)R1’, or P(O2)R1’; R1’ is C1-6alkyl, C2-6alkenyl, C1-6alkylene-5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S, C3-10cycloalkyl, C6-10aryl, 5- to 10 membered heterocycloalkyl comprising one to threeheteroatoms selected from O, N, and S, or 5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S; and Ra’ and Rb’ are individualy H, C1-6alkyl, C2-6alkenyl, C(O)OC1-6alkyl, C3-10cycloalkyl, C6-10aryl, 5- to 10 membered heterocycloalkyl comprising one to three heteroatoms selected from O, N, and S, or 5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S, or C1-6alkylene-5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S; the method comprising a cyclization step comprising admixing a compound of Formula (a) with (Z)-1- cyclopropyl-3-(dimethylamino)-2-(3-methylisoxazol-5-yl)prop-2-en-1-one (“Compound 12”) in the presence of a base and a polar solvent: ,reaction between the compound of Formula (a) and Compound 12.

39. The process of claim 38, wherein R’ is C1-6alkylene-5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S.

40. The process of claim 38, wherein R’ is C2alkylene-5-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S.

41. The process of any one of claims 38 to 40, wherein the compound of Formula (a) is .of any one of claims 38 to 41, wherein the compound of Formula (I) has the structure of Compound 1: .of claim 38, wherein R’ is OR1’, SR1’, or NR’aRb’.

44. The process of claim 43, wherein R’ is OR1’ or SR1’.

45. The process of claim 43 or 44, wherein R1’ is C1-6alkyl.

46. The process of any one of claims 43 to 45, wherein R1’ is methyl.

47. The process of claim 38, wherein R is NRa’Rb’.

48. The process of any one of claims 38, 43, or 47, wherein Ra’ and Rb’ are individualy H, C1-6alkyl, C2- 6alkenyl, C(O)OC1-6alkyl, C3-10cycloalkyl, C6-10aryl, 5- to 10 membered heterocycloalkyl comprising one to three heteroatoms selected from O, N, and S, or 5- to 8-membered heteroaryl comprising one to three heteroatoms selected from O, N, and S.

49. The process of claim 48, wherein Ra’ is H.

50. The process of claim 48 or 49, wherein Rb’ is H.

51. The process of claim 48 or 49, wherein Rb’ is C1-6alkyl or C(O)OC1-6alkyl.

52. The process of claim 51, wherein Rb’ is C1-6alkyl.

53. The process of claim 52, wherein Rb’ is methyl.

54. The process of claim 51, wherein Rb’ is C(O)OC1-6alkyl.

55. The process of claim 54, wherein Rb’ is C(O)O-t-butyl.

56. The process of any one of claims 38 to 55, wherein prior to the cyclization step the process comprises: (i) a condensation step comprising admixing 3,5-dimethylisoxazole and methyl cyclopropane carboxylate in the presence of a base in a polar solvent, under conditions suficient to efect a condensation reaction to yield 1- cyclopropyl-2-(3-methylisoxazol-5-yl)ethan-1-one (“Compound 11”): admixing Compound 11 and N,N-dimethylformamide dimethyl acetal (DMF-DMA) under conditions suficient to form Compound 12.

57. The process of claim 56, wherein the condensation step (i) comprises admixing 3,5- dimethylisoxazole and methyl cyclopropane carboxylate in the presence of a base in a polar aprotic solvent.

58. The process of claim 56, wherein the polar aprotic solvent is tetrahydrofuran (THF).

59. The process of any one of claims 56 to 58, wherein the base is lithium disopropylamide (LDA).

60. The process of claim 57 or 58, wherein the admixing is caried out at a temperature of - 70 °C or lower.

61. The process of claim 57 or 58, wherein the admixing is caried out at a temperature of about - 78 °C.

62. The process of any one of claims 56 to 60, wherein the condensation step (i) comprises the steps of: (ia) combining 3,5-dimethylisoxazole and THF to form a first solution; (ib) admixing the first solution with LDA that has been pre-cooled to a temperature of - 70 °C or lower to form a second solution; and(ic) admixing the second solution with methyl cyclopropane carboxylate under conditions suficient to yield Compound 11.

63. The process of claim 62, wherein the methyl cyclopropane carboxylate is provided as a solution of methyl cyclopropane carboxylate in THF.

64. The process of claim 62 or 63, wherein the LDA has been pre-cooled to a temperature of about - 78 °C.

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