Neuroprotective compounds and methods of use

Abstract

Provided herein are compounds of formula (I), or a pharmaceutically acceptable salt thereof: (I) wherein A1, A2, R1, and R2 are as described herein. Further provided are pharmaceutical compositions of (I) and methods of using the disclosed compounds, e.g., treating diseases, disorders, or conditions associated with, or caused by or associated with mitochondrial degradation (e.g., neurodegenerative diseases).

Description

S-T00387W0001 32917 / 70090 / PCNEUROPROTECTIVE COMPOUNDS AND METHODS OF USE FUNDING ACKNOWLEDGEMENT STATEMENT

[0001] This invention was made with government support under NS097224 and R33 AG068887 awarded by the National Institutes of Health. The government has certain rights in the invention. The invention was created in whole or in part by funding received from the Lottie French Lewis Fund of the Community Foundation for Palm Beach and Martin Counties.BACKGROUND

[0002] Neurodegenerative diseases and disorders, such as Alzheimer's disease (AD), Parkinson's disease (PD), and amyotrophic lateral sclerosis (ALS), have a profound impact on society. These conditions lead to the progressive loss of cognitive and motor functions, resulting in severe disability, a decline in quality of life, and eventual death. The societal impact includes emotional and financial strain on families, increased demand for healthcare and long-term care services, and significant economic costs due to lost productivity and medical expenses. As the population ages, the prevalence of these diseases is projected to rise, posing further challenges for healthcare systems and communities worldwide.

[0003] ALS is a rare neurodegenerative disease, affecting between 5-10 individuals per 100,000. (Arthur et al., 2016; Mehta et al., 2018; Mehta et al., 2022). ALS comes in both familial and sporadic forms (fALS and sALS; Brown et al., 2017; Hardiman et al., 2017; Mathis et al., 2019; van Rheenen et al., 2021). The most frequent form of fALS is due to repeat expansion mutations in the C9orf72 gene (approximately 40% of cases). Other genetic forms of fALS include mutations in the superoxide dismutase 1 (SOD1) gene and the TAR-DNA binding protein 43 (TDP43) gene. These other genetic forms represent about 20% and 5% of the heritable cases, respectively. More than 30 other genes contribute to the remainder of fALS cases. The heritable cases of ALS comprise about 10% of all ALS cases, with 90% due to unknown causes and thus categorized as sporadic ALS.

[0004] The three genetic forms of ALS - C9orf72, SOD1, and TDP43 - represent very distinct causes for ALS. The gene C9orf72 is thought to encode a protein involved in lysosomal function. The SOD1 gene encodes a protein that decreases oxidative stress. The TDP43 gene encodes a protein that binds to DNA and RNA and alters gene and protein expression. The general molecular / cellular and organismal pathology observed in ALS patients is similar despite the distinct insults due to mutations in these three different genes.

[0005] The age of onset of sALS is approximately 55-65 years of age, whereas fALS occurs in the late teen or adult years. ALS patients usually die within 2-5 years of diagnosis, typically from respiratory paralysis. ALS represents a disease spectrum, including fALS, fFTD (frontotemporal dementia), and sALS, with different initial causes illustrated by the distinct gene mutations that cause fALS / fFTD but overlapping neuropathology (Figure 1). Although many factors can trigger disease onset and progression, the molecular I cellular neuropathology across the spectrum remains generally constant. ALS and many neurodegenerative diseases such as AD canS-T00387W0001 32917 / 70090 / PCbe characterized by early mitochondrial (MT) dysfunction, which triggers a cascade of molecular and cellular damage.

[0006] Thus, there remains a need for compounds for treating neurodegenerative diseases and disorders, including ALS and other diseases characterized by MT dysfunction and neurodegeneration.SUMMARY

[0007] The disclosure provides compounds of formula (I):A1r"n.' ' -R2N A2(|), or pharmaceutically acceptable salts thereof, whereinA1is selected from morpholinyl substituted with 0-2 substituents independently selected from Ci-3alkyl and Ci-shydroxyalkyl; piperidinyl substituted with 0-2 substituents independently selected from Ci-salkyl, hydroxy, and Ci-shydroxyalkyl; and -N(R3)2;A2is selected from piperidinyl, pyrrolidinyl, azetidinyl, -N(R4)2, -O-Ci-shydroxyalkyl, -S-Ci-shydroxyalkyl, halo, and morpholinyl substituted with 0-2 substituents independently selected from Ci-3alkyl, and Ci-shydroxyalkyl, wherein the piperidinyl, and pyrrolidinyl are substituted with 0-3 substituents independently selected from Ci-salkyl, Ci-salkoxy, hydroxy, Ci-shydroxyalkyl, phenyl, and halo;A1or A2optionally further comprise a labile linker;R1and R2are each independently N- or CH, wherein R1and R2together with carbon and nitrogen atoms to which they are attached form a 5- or 6-membered ring, having 0-2 ring N atoms, and the 5- or 6-membered ring is substituted with 0-2 substituents independently selected from A3and -Ci-salkylene-O-Ci-salkyl;each A3is independently selected from morpholinyl substituted with 0-2 substituents independently selected from Ci-salkyl, and Ci-shydroxyalkyl; piperidinyl, pyrrolidinyl, -N(R5)2, -Ci-shydroxyalkyl, and -Ci. salkylene-Ci-salkoxy, wherein the piperidinyl and pyrrolidinyl are substituted with 0-2 substituents independently selected from Ci-salkyl, hydroxy, and Ci-shydroxyalkyl;each R3is independently Ci-ealkyl or Ci-shydroxyalkyl; andeach R4and R5is independently selected from H, Cvealkyl, Ci-ehydroxyalkyl, and Ci-salkyl-O-Ci.ealkylene; with the proviso that the compound is notorS-T00387W0001 32917 / 70090 / PC, and at least one of A1, A2, and A3is piperidinyl or pyrrolidinyl optionally substituted with 0-3 substituents independently selected from Ci-salkyl, Cvsalkoxy, hydroxy, Ci-shydroxyalkyl, phenyl, and halo, or at least one of A1, A2, and A3is morpholinyl optionally substituted with 0-2 substituents independently selected from C^alkyl and Ci-shydroxyalkyl, or at least one R4or R5is Ci-ehydroxyalkyl, and Ci.3alkyl-O-Ci-6alkylene

[0008] The disclosure also provides pharmaceutical compositions comprising the disclosed compounds and pharmaceutically acceptable salts, as well as methods of treating neurodegenerative diseases and disorders.BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 shows mutant genes (> 30) that cause familial forms of FTD and ALS and their overlap. Adapted from Gao et al., 2017.

[0010] Figure 2 shows the MT cascade hypothesis. Multiple etiologies for ALS may initiate a cascade of neuropathology, with the emergence and progression of MT dysfunction as an important upstream event, with the final endpoint being the death of neurons in the brain and spinal cord. The cascade is illustrated as parallel arms from MT dysfunction, although some downstream aspects of neuropathology could occur in a serial arrangement.

[0011] Figure 3 shows the structure of dipyridamole (DPM).

[0012] Figures 4A-4C show C9orf72 (C9) IPSC-derived motor neurons (MNs) treated with DPM at day 12. These DPM-treated C9 MN show extended signs of general and MT health over a thirty-day time course, with DMSO-treated MN generally dying by day 18-20. The DMSO-treated C9 MN show strong, reproducible, and progressive MT dysfunction phenotypes (reduced number, shortening) and also a neurite degeneration phenotype, as compared to isogenic (ISO) MN across days in culture. The DPM-treated C9 MN show rescued MT and neurite phenotypes. CA=Cumulative Area, the average number of pixels representing neurites in the images. Data are line graphs from five independent time course experiments with DPM at 10 m added at day 12. [Black asterisk C9 MN (DMSO) vs ISO MN (DMSO); gray asterisk C9 MN (DPM) vs 09 MN (DMSO). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, two-way repeated measure ANOVA, Tukey's multiple comparisons test (n=5 independent experiments)].

[0013] Figures 5A and 5B show representative images from day 30 for DMSO or DPM-treated 09 MN, showing the neuroprotective effect of DPM. In these images, the MT are highlighted with a MT-targeted Green fluorescent protein (GFP) reporter (MT-GFP), while the MN cell bodies and neurites (axons and dendrites) are highlighted with a cytoplasmically localized fluorescent mScarlet reporter. MT-GFP panels illustrate the labeledS-T00387W0001 32917 / 70090 / PCMT (DPM) or MT remnants (DMSO) while mScarlet panels illustrate protected (DPM) or degenerated (DMSO) MN axons and dendrites. Note especially the integrity of MT (points on a string) in DPM-treated C9 MN. Large objects are somas or cellular debris.

[0014] Figure 6 shows dose-dependent maintenance of cell viability in DPM-treated C9 MN. The assay was run in a dose:response (D: R) format with the RealTime-Glo MT Cell Viability reagent. Data are box plots from three independent experiments, with the reagent added at day 31 and viability assayed at day 36. DPM concentrations of 5-20 piM dramatically increase the survival of the C9 MN. [Black asterisk ISO MN (DMSO) vs 09 MN (DMSO) and some groups of 09 MN (DPM); gray asterisk 09 MN (DMSO) vs some groups of 09 MN (DPM). ****p<0.0001, One-way ANOVA, Tukey's multiple comparisons test (n=3 independent experiments)].

[0015] Figures 7A, 7B, 70, and 7D (Top of each panel) illustrate dose:response assays for oxygen consumption (basal, ATP-linked, and maximal respiration; and spare respiratory capacity) in isogenic (ISO, gray) and 09 ALS (black) motor neurons (MN), both neuron types treated with DPM. The data show a deficit in the four respiration parameters without DPM (0 concentration, DMSO-treated, gray asterisks), and that DPM added in increasing dosage improves the 09 MN respiration to a level significantly above the level without DPM (black asterisks) and to the level of the ISO MN. This illustrates that the DPM bring MT bioenergetics back to normal levels. [Gray asterisk 09 MN (DMSO) vs ISO MN (DMSO); black asterisk 09 MN (DPM) vs 09 MN (DMSO). *p<0.05, **p<0.01, two-way ANOVA, Sidak's multiple comparisons test (n=3 independent experiments)]. (Bottom of each panel) Dose:response curves for the effect of DPM on each parameter of respiration. DPM's EC50 values for these four MT functional parameters are in the sub-micromolar range, indicating its high potency in restoring MT function.

[0016] Figures 8A-8C show representative D: R curves for DPM at day 20 for MT count, MT length, and neurite CA using 09 MN. Data are shown only for day 20. Measured ECso's and spans for the 3 parameters are also in Table 1. Note the difference in y:axis scales. Potency is measured by EC50 and efficacy by span (the y-axis difference between the baseline and asymptotic Robust Z-score).

[0017] Figure 9 shows how a representative analog of DPM (DPM-A19) improves the MT length parameter over DPM. Another analog DPM A-33 is inactive and behaves like DMSO. Data are line graphs from a time course experiment with the compound at 10 m added at day 12. [gray asterisk 09 MN (analog or DPM) vs 09 MN (DMSO); black asterisk 09 MN (analog) vs 09 MN (DPM); *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Two-way repeated measure ANOVA, Tukey's multiple comparisons test.

[0018] Figures 10A-10C show D: R curves for a previously unreported compound (DPM-A19) that is structurally related to DPM (top panel) compared to analogous curves for DPM (bottom panel). The D: R curves show that the previously DPM analog A19 has improved potency (EC50) and efficacy (Span, the y-axis difference between baseline and the asymptotic value of the curve) for the three parameters studied, MT count (10A), MT length (10B), and neurite CA (10C). Data are shown only for day 18.S-T00387W0001 32917 / 70090 / PC

[0019] Figures 11 A, 11 B, and 11C (top panels) show data obtained in a time course assay of MT count, MT length, and Neurite CA (Cumulative Area of pixels representing axons and dendrites), respectively, using AD Presenilin 1 (PS1-A246E) IPSC-derived cortical neurons (CN) without compound (DMSO, black) or with DPM (gray, closed circle) or two DPM analogs: DPM-A7b (gray open circle), and DPM-A19 (gray open square). This shows that DPM and analogs dramatically improve MT count, length, and axon / dendrite integrity. [Black asterisk PS1-A246E CN (DPM) vs PS1-A246E CN (DMSO); gray asterisk PS1-A246E CN (DPM-A7b) vs PS1-A246E CN (DMSO); gray hashtag PS1-A246E CN (DPM-A19) vs PS1-A246E CN (DMSO). **p<0.05, **»p<0.01, ***p<0.001, ****p<0.0001, two-way ANOVA, Dunnett's post hoc test], (Bottom panels) The respective EC50's and spans for DPM-A19 as determined by dose:response assays at day 20 of the time course data are illustrated in the bottom panels of each figure. The compounds tested promote all three parameters with potency of a representative compound between 0.7 and 1.8 μM relative to AD IPSC-derived neurons treated with DMSO control. These data demonstrate that the compounds offset the loss of MT content, MT fragmentation, and axon / dendrite degeneration due to the disease.

[0020] Figures 12A, 12B, 12C, and 12D (top of each panel) illustrate dose:response assays for oxygen consumption (basal, ATP-linked, and maximal respiration; and spare respiratory capacity) in isogenic (ISO, gray) and AD PS1 (PS1-A246E, black) cortical neurons (CN), both neuron types treated with DPM. The data show a deficit in the four respiration parameters without DPM (0.0 concentration, gray asterisks), and that DPM added in increasing dosage improves the AD neuron respiration to a level significantly above the level without DPM (black asterisks) and to the level of the ISO neurons. This illustrates that DPM bring MT bioenergetics back to normal levels. [Gray asterisk PS1-A246E CN (DMSO) vs ISO CN (DMSO); black asterisk PS1-A246E CN (DPM) vs ISO CN (DMSO). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, two-way ANOVA, Sidak's multiple comparisons test (n=3 independent experiments)]. (Bottom of each panel) Dose:response curves for the effect of DPM on each parameter of respiration.

[0021] Figure 13 A-C (Top of each panel) Measures obtained in a time course assay of MT count (Panel A), MT length (Panel B), and Neurite CA (Panel C; Cumulative Area of pixels representing axons and dendrites) using ALS mutant TDP43 (M337V) IPSC-derived motor neurons (MN) without (DMSO, gray filled circles) or with DPM (gray open circles). This shows that DPM dramatically improves MT count, MT length, and axon / dendrite integrity in this model of TDP43 ALS so that they are close to the non-mutant (Control, black filled circles) values.[Black asterisk TDP43M337VMN (DMSO) vs Control MN (DMSO); gray asterisk TDP43M337VMN (DPM) vs TDP43M337VMN (DMSO). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001]. Two-way repeated measure ANOVA, Tukey's multiple comparisons test (n=5 independent experiments)]. (Bottom of each panel) The EC50's and spans as determined by dose:response assays at day 20 of the time course data are illustrated for the top graphs.

[0022] Figures 14A, 14B, and 14C (top panels) show data obtained in a time course assay of MT count, MT length, and Neurite CA (Cumulative Area of pixels representing axons and dendrites) using ALS mutant ALS-SOD1 (A5V) IPSC-derived motor neurons (MN) without (DMSO, solid gray circles) or with DPM (open grayS-T00387W0001 32917 / 70090 / PCcircles), or the ISO control (black circles). This shows that DPM dramatically improves MT count, MT length, and axon / dendrite integrity so that they are close to the control values. [Black asterisk SOD1 (A5V) MN (DMSO) vs ISO MN (DMSO); gray asterisk SOD1 (A5V) MN (DPM) vs SOD1 (A5V) MN (DMSO)]. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, two-way ANOVA with Dunnett's post hoc test]. The respective EC50's and spans (Bottom panels) as determined by dose:response assays at day 22 of the time course data are illustrated.

[0023] Figure 15 illustrates cartoon of mitochondria showing that the uptake of pyruvate, which is derived from glucose metabolism, is mediated by the Mitochondrial Pyruvate Carrier (MPG) protein.

[0024] Figure 16 illustrates results of measuring pyruvate uptake into isolated mouse liver mitochondria. The well-characterized inhibitor of MPC (UK5099) inhibits pyruvate uptake in a dose-dependent manner. DPM and a DPM analog A23 augment pyruvate uptake into isolated mitochondria. Each point within each bar indicates results from an independent experiment (n = 3-5 independent experiments). Asterisks in each of the 3 bar graph groups indicate significant differences from its control with no (zero) compound.] *p<0.05, **p<0.01, and ****p<0.0001, mixed model ANOVA with Student's t post hoc test].DETAILED DESCRIPTION

[0025] The present disclosure provides compounds useful for treating diseases and disorders characterized by degradation of MT (e.g., neurodegenerative diseases and disorders). MT play a central role in maintaining the health of neurons as the powerhouse for energy in the form of adenosine triphosphate (ATP) and participate in many other essential aspects of cellular physiology as well. Provided herein are molecules that delay and / or markedly reduce the progression of MT dysfunction during disease progression. Improved MT health helps to maintain axons and dendrites of motor neurons thereby protecting the motor neurons from degradation and death.

[0026] In some embodiments, the present disclosure provides compounds of formula (I), or pharmaceutically acceptable salt salts thereof:A1R1 USRwherein A1, A2, R1, and R2are as defined herein, with the proviso that the compound is notOHN^N XNZNZAN OH, and at least one of A1, A2, and A3is piperidinyl or pyrrolidinyl optionally substituted with 0-3 substituents independently selected from Ci-salkyl, Cvsalkoxy, hydroxy, Ci-shydroxyalkyl, phenyl, and halo, or at least one of A1, A2, and A3is morpholinyl optionally substituted with 0-2S-T00387W0001 32917 / 70090 / PCsubstituents independently selected from Ci-salkyl and Ci-shydroxyalkyl, or at least one R4or R5is Ci. ehydroxyalkyl, and Ci-3alkyl-O-Ci-ealkylene

[0027] In some embodiments, the disclosure provides the disclosed compounds in the form of a pharmaceutically acceptable salt. The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness and properties of the compound and which typically is not biologically or otherwise undesirable. In many cases, the compounds of the present disclosure are capable of forming acid and / or base salts by virtue of the presence of amino and / or carboxyl groups or groups similar thereto.

[0028] Pharmaceutically acceptable acid addition salts are known in the art and can be formed with inorganic (mineral) acids and I or with organic acids. Inorganic acids from which salts can be derived include, as noncomprehensive examples, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Organic acids from which salts can be derived include, as noncomprehensive examples, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, benzenesuflonic acid, toluenesulfonic acid, and sulfosalicylic acid.

[0029] Pharmaceutically acceptable base addition salts are known in the art can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, as noncomprehensive examples, ammonium salts and metals from columns I to XII of the periodic table. In certain embodiments, the salts are derived from (as noncomprehensive examples) sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper. In certain other embodiments, the salts are selected from ammonium, potassium, sodium, calcium and magnesium salts. Organic bases from which salts can be derived include, as examples, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like. Certain organic amines include isopropylamine, benzathine, cholinate, diethanolamine, diethylamine, lysine, meglumine, piperazine and tromethamine.

[0030] The compounds of the disclosure are defined herein by their chemical structures and / or chemical names. For avoidance of doubt, wherein a compound is referred to by both a chemical structure and a chemical name, and the chemical structure and chemical name conflict, the chemical structure is determinative of the compound's identity.Chemical Definitions

[0031] As used herein, the term "halo” or "halogen" refers to fluoro, chloro, bromo, and / or iodo.

[0032] As used herein, the term "alkyl" generally refers to a fully saturated branched or unbranched hydrocarbon moiety having up to 20 carbon atoms. Unless otherwise provided, alkyl refers to hydrocarbon moieties having 1 to 20 carbon atoms, 1 to 16 carbon atoms, 1 to 10 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms. More particularly, the term Cnmeans the alkyl group has “n” carbon atoms. For example, C4 alkyl refers to an alkyl group that has 4 carbonS-T00387W0001 32917 / 70090 / PCatoms. Accordingly, the term Ci-salkyl refers to an alkyl group having a number of carbon atoms encompassing the entire range (e.g., 1, 2, and / or 3 carbon atoms). Suitable representative examples of Ci-salkyl groups include, methyl, ethyl, n-propyl, and isopropyl. Similarly, Ci-salkyl refers to an alkyl group having a number of carbon atoms encompassing the entire range (e.g., 1 to 8 carbon atoms), as well as all subgroups (e.g., 1-8, 2-8, 3-8, 4-8, 5-8, 6-8, 7-8, 1, 2, 3, 4, 5, 6, 7, and 8 carbon atoms). Nonlimiting examples of alkyl groups include, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl (2-methylpropyl), t-butyl (1,1 -dimethylethyl), 3,3-dimethylpentyl, and 2-ethylhexyl.

[0033] As used herein, the term " Cx-yhydroxyalkyl” refers to a Cx-yalky I moiety that is substituted with one or more hydroxy groups on the alkyl chain.

[0034] As used herein, the term "alkoxy” refers to -OR, wherein R is alkyl as described herein. Illustrative examples of Ci-3alkoxy include methoxy, ethoxy, n-propoxy, and isopropoxy.

[0035] As used herein, the term "alkylene" refers to divalent alkyl group as defined herein above having 1 to 20 carbon atoms. Unless otherwise provided, alkylene refers to moieties having 1 to 20 carbon atoms, 1 to 16 carbon atoms, 1 to 10 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms. The term Cnmeans the alkylene group has “n” carbon atoms. For example, C2alkylene refers to an alkylene group that has 2 carbon atoms. Ci-2alkylene refers to an alkylene group having a number of carbon atoms encompassing the entire range (e.g., 1 to 2 carbon atoms). Moreover, the term Coalkylene refers to 0 carbon atoms, that is, the Coalkylene moiety is a direct bond. Representative examples of alkylene include, but are not limited to, methylene, ethylene, n-propylene, iso-propylene, n-butylene, sec-butylene, iso-butylene, tert-butylene, n-pentylene, isopentylene, neopentylene, n-hexylene, 3-methylhexylene, 2,2-dimethylpentylene, 2,3-dimethylpentylene.

[0036] As used herein, the term "optionally substituted" unless otherwise specified refers to a group that is unsubstituted or is substituted with one or more, typically 1, 2, 3, or 4, suitable non-hydrogen substituents. If the identity of the "optional substituent” is not clearly defined in context of the optionally substituted group, then each optional substituent is independently selected from the group consisting of: alkyl, hydroxy, halogen, oxo, amino, alkylamino, dialkylamino, alkoxy, cycloalkyl, CO2H, heterocycloalkyloxy (which denotes a heterocyclic group bonded through an oxygen bridge), -CC^alkyl, mercapto, nitro, cyano, sulfamoyl, sulfonamide, aryl, -OC(O)alkyl, -OC(O)aryl, aryl-S-, aryloxy; alkylthio, formyl (i.e., HC(O)-), -C(O)NH2, aralkyl (alkyl substituted with aryl), aryl and aryl substituted with alkyl, cycloalkyl, alkoxy, hydroxy, amino, alkyl-C(O)— NH—, alkylamino, dialkylamino or halogen. It is understood that where a group is indicated to be optionally substituted, the disclosure includes embodiments in which the group is unsubstituted as well as embodiments in which the group is substituted.

[0037] As used herein, the term "phenyl” refers to substituted or unsubstituted -CeHs.

[0038] As used herein, the term "azetidinyl" refers to -1-.S-T00387W0001 32917 / 70090 / PC

[0039] Compounds of the disclosure can be in the form of one of the possible isomers, rotamers, atropisomers, tautomers or mixtures thereof, for example, as substantially pure geometric (cis or trans) isomers, diastereomers, optical isomers (antipodes), racemates or mixtures thereof. It is understood that for any compound provided herein, including any compound of formula (I), or any embodiment thereof, or a salt of any of the foregoing, the compound may exist in any stereochemical form, such as a single enantiomer, diastereomer, or tautomer or a mixture of one or more enantiomers, diastereomers, and tautomers in any ratio.

[0040] For example, any asymmetric atom (e.g., carbon or the like) of the compound(s) of the present invention can be present in racemic or enantiomerically enriched, for example the (R)-, (S)- or (Reconfiguration. In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess (ee), at least 60% ee, at least 70% ee, at least 80% ee, at least 90% ee, at least 95% ee, or at least 99% ee in the (R)-or (S)-configuration. Substituents at atoms with unsaturated bonds may, if possible, be present in cis-(Z)- or trans-(E)-form.

[0041] As used herein, the term "isomers" refers to different compounds that have the same molecular formula but differ in arrangement and configuration of the atoms. Also as used herein, the term "an optical isomer" or "a stereoisomer" refers to any of the various stereo isomeric configurations which may exist for a given compound of the present invention and includes geometric isomers. It is understood that a substituent may be attached at a chiral center of a carbon atom. Therefore, the invention includes enantiomers, diastereomers or racemates of the compound. " Enantiomers" are a pair of stereoisomers that are non-superimposable mirror images of each other. A 1: 1 mixture of a pair of enantiomers is a "racemic" mixture. The term is used to designate a racemic mixture where appropriate. The use of "rel" indicates that the diastereomeric orientation is known but the absolute stereochemistry is not. In cases where the absolute stereochemistry has not been determined the optical rotation and / or chiral chromatography conditions will indicate which isomer is present.

[0042] " Diastereoisomers" are stereoisomers that have at least two asymmetric atoms, but which are not mirror-images of each other. The absolute stereochemistry is specified according to the Cahn-I ngold-Prelog R-S system. When a compound is a pure enantiomer the stereochemistry at each chiral carbon may be specified by either R or S. Resolved compounds whose absolute configuration is unknown can be designated (+) or (-) depending on the direction (dextro- or levorotatory) which they rotate plane polarized light at the wavelength of the sodium D line or retention time on chiral chromatography separation. Certain of the compounds described herein contain one or more asymmetric centers or axes and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)-, or with the (+) or (-) sign. The present invention is meant to include all such possible isomers, including racemic mixtures, optically pure forms and intermediate mixtures. Optically active (R)- and (S)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If the compound contains aS-T00387W0001 32917 / 70090 / PCdouble bond, the substituent may be E or Z configuration. If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have a cis- or trans-configuration.

[0043] Any resulting mixtures of isomers can be separated on the basis of the physicochemical differences of the constituents, into the pure or substantially pure geometric or optical isomers, diastereomers, racemates, for example, by chromatography and / or fractional crystallization.

[0044] Any resulting racemates of final products or intermediates can be resolved into the optical antipodes by known methods, e.g., by separation of the diastereomeric salts thereof, obtained with an optically active acid or base, and liberating the optically active acidic or basic compound. In particular, a basic moiety may thus be employed to resolve the compounds of the present invention into their optical antipodes, e.g., by fractional crystallization of a salt formed with an optically active acid, e.g., tartaric acid, dibenzoyl tartaric acid, diacetyl tartaric acid, di-O. O'-p-toluoyl tartaric acid, mandelic acid, malic acid or camphor-10-sulfonic acid. Racemic products can also be resolved by chiral chromatography, e.g., high performance liquid chromatography (HPLC) or supercritical fluid chromatography (SFC) using a chiral adsorbent.

[0045] Mixtures of isomers obtainable according to the invention can be separated in a manner known to those skilled in the art into the individual isomers; diastereoisomers can be separated, for example, by partitioning between polyphasic solvent mixtures, recrystallization and / or chromatographic separation, for example over silica gel or by e.g. medium pressure liquid chromatography over a reversed phase column, and racemates can be separated, for example, by the formation of salts with optically pure salt-forming reagents and separation of the mixture of diastereoisomers so obtainable, for example by means of fractional crystallization, or by chromatography over optically active column materials. Certain compounds disclosed herein are named using pseudoasymmetric descriptors (r / s) by compound-naming programs such as ChemDraw (e.g., see Stereochemistry of organic compounds by E. L. Eliel and S. H. Wilen. John Wiley & Sons: 1994, pages 666-668.) Compounds of Formula (I)A1, A2, R3, and R4A1R1 US2

[0046] Compounds of formula (I): '' R (I) comprise an A1moiety selected from morpholinyl substituted with 0-2 substituents independently selected from Ci-salkyl and Ci-shydroxyalkyl; piperidinyl substituted with 0-2 substituents independently selected from Ci-salkyl, hydroxy, and Ci-shydroxyalkyl; and -N(R3)2, wherein each R3is independently Cvealkyl or Ci-shydroxyalkyl.S-T00387W0001 32917 / 70090 / PC

[0047] In some embodiments, A1is selected from selected from-L, -L, ~1~, J-, -L, -1-, andu O 0

[0048] In some embodiments, A1is selected from J-, -L, J~, and O --J-'O O Od O 0

[0049] In some embodiments, A1is selected from -J-, J—, and -L-.

[0050] Compounds of formula (I) comprise A2is selected from piperidinyl, pyrrolidinyl, azetidinyl, -N(R4)2, -O- Ci-shydroxyalkyl, -S-Ci-shydroxyalkyl, halo, and morpholinyl substituted with 0-2 substituents independently selected from Ci-salkyl, and Ci-shydroxyalkyl, wherein the piperidinyl, and pyrrolidinyl are substituted with 0-3 substituents independently selected from Ci-salkyl, Cvsalkoxy, hydroxy, Ci-shydroxyalkyl, phenyl, and halo;

[0051] In some embodiments, A2is selected from piperidinyl, pyrrolidinyl, -N(R4)2, -O-Ci-ahydroxyalkyl, -S-Ci. shydroxyalkyl, halo and morpholinyl substituted with 0-2 substituents independently selected from Ci-3alkyl, and Ci-shydroxyalkyl, wherein the piperidinyl, and pyrrolidinyl are substituted with 0-2 substituents independently selected from Ci-3alkyl, hydroxy, and Ci-shydroxyalkyl, and each R4is independently selected from H, Ci-ealkyl, Ci-ehydroxyalkyl, and Ci-salkyl-O-Ci-ealkylene.

[0052] In some embodiments, A2is selected from azetidinyl, Cvsalkoxy, phenyl, and halo.

[0053] In some embodiments, A2is substituted with 0-3 substituents (e.g., 0, 1, 2, or 3), as described herein.

[0054] In some embodiments, A2is substituted with 0-2 substituents (e.g., 0, 1, or 2), as described herein.S-T00387W0001 32917 / 70090 / PC

[0056] In some embodiments, A2is selected from — I—, — J—. -L, _L, j_, ™LLabile Linkers

[0058] In some embodiments, A1or A2comprises a labile linker. As used herein "labile linker” refers to a chemical moiety that is designed to be unstable or prone to degradation under specific physiological conditions, such as changes in pH, enzymatic activity, or reducing environments. In some embodiments, the labile linker comprises a functional group selected from a hydrazone, a disulfide, an ester, a carbamate, a peptide, an azo, and an oxime, attached covalently to corresponding groups present in A1or A2in methods well understood by persons skilled in the art. In some embodiments, the labile linker is comprises a phosphate or phosphonate ester, sulfate or sulfonic ester, or a carboxylic ester, wherein the ester comprises an alkyl or amino acid residue.R1, R2, A3, and R5

[0059] Compounds of formula (I) comprise R1and R2moieties, wherein each are independently N- or -CH, or R1and R2together with carbon and nitrogen atoms to which they are attached form a 5- or 6-membered ring, having 0-2 ring N atoms, and the 5- or 6-membered ring is substituted with 0-2 substituents independently selected from A3and -Ci-salkylene-O-Ci-salkyl. In some embodiments, at least one R1and R2is Ci-salkyl.

[0060] Each A3moiety is independently selected from morpholinyl substituted with 0-2 substituents independently selected from Ci-salkyl, and Ci-shydroxyalkyl; piperidinyl, pyrrolidinyl, -N(R5)2, -Ci-shydroxyalkyl, and -Ci-salkylene— Ci-salkoxy, wherein the piperidinyl and pyrrolidinyl are substituted with 0-2 substituents independently selected from Ci-salkyl, hydroxy, and Ci-shydroxyalkyl, and each R5is independently selected from H, Ci-ealkyl, Ci-ehydroxyalkyl, and Ci-salkyl-O-Ci-ealkylene.S-T00387W0001 32917 / 70090 / PCCompounds of Formula (l-A)

[0061] In some embodiments, the disclosure provides compounds of formula (l-A): wherein each X1is independently N or CH. In some embodiments, at least one X1is N. In some embodiments,A1at least one X1is CH. In some embodiments, the compound of formula (l-A) is A1orCompounds of Formula (l-B)

[0062] In some embodiments, the present disclosure provides compounds of formula (l-B)A6X2-^^NA5~{ T JN N^ TK2A4(|-B), wherein X2is N or CH; and A4, A5, and A6are each independently H, C1-4alkyl, morpholinyl substituted with 0-2 substituents independently selected from Ci-3alkyl, and Ci-shydroxyalkyl, piperidinyl, pyrrolidinyl, -N(R5)2, Ci-shydroxyalkyl, and Ci-3alkylene-O-Ci-3alkyl, wherein the piperidinyl and pyrrolidinyl are substituted with 0-2 substituents independently selected from Ci-3alkyl, hydroxy, and Ci-shydroxyalkyl.

[0063] In some embodiments, X2of formula (l-B) is N. In some embodiments, X2of formula (l-B) is CH.

[0064] In some embodiments, A4of formula (l-B) is H. In some embodiments, A4of formula (l-B) is Ci-4alkyl.

[0065] In some embodiments, the present disclosure provides a compound selected fromS-T00387W0001 32917 / 70090 / PCS-T00387W0001 32917 / 70090 / PC DPM-A20,S-T00387W0001 32917 / 70090 / PCS-T00387W0001 32917 / 70090 / PCDPM-A61.

[0066] In some embodiments, the present disclosure provides a compound selected fromS-T00387W0001 32917 / 70090 / PCS-T00387W0001 32917 / 70090 / PCS-T00387W0001 32917 / 70090 / PC OH DPM-A41,

[0067] In some embodiments, the present disclosure provides a compound selected fromS-T00387W0001 32917 / 70090 / PC

[0068] In some embodiments, the present disclosure provides a compound selected from DPM-A2, DPM-A6, DPM-A10, DPM-A11, DPM-A13, DPM-A17, DPM-A19, DPM-A20, DPM-A21, DPM-A22, DPM-A23, DPM-A24, DPM-A28, DPM-A36, DPM-A35, DPM-A34, DPM-A39, DPM-A40, DPM-A46, DPM-A47, DPM-A41, DPM-A42, DPM-A43, DPM-A44, DPM-A45, DPM-A48, DPM-A49, DPM-A50, DPM-A52, DPM-A54, and DPM-A55.

[0069] In some embodiments, the present disclosure provides a compound selected from DPM-A43, DPM-DPM-A44, DPM-A45, DPM-A46, DPM-A47, DPM-A48, DPM-A49, DPM-A50, DPM-A51, DPM-A52, DPM-A53, DPM-A54, DPM-A55, DPM-A56, DPM-A57, DPM-A58, DPM-A59, DPM-A60, and DPM-A61.Pharmaceutical Compositions

[0070] In some embodiments, the present disclosure provides pharmaceutical compositions comprising the disclosed compounds (e.g., compounds of formula (I), (l-A), and (l-B)) and a pharmaceutically acceptable carrier.S-T00387W0001 32917 / 70090 / PCIn some embodiments, the disclosed pharmaceutical compositions comprise a pharmaceutically acceptable salt of the disclosed compounds and a pharmaceutically acceptable carrier.

[0071] In some embodiments, the pharmaceutical composition is formulated for oral, parenteral, inhalational, intranasal, subcutaneous, intramuscular, or intravenous administration. In some embodiments, the pharmaceutical composition is formulated for oral administration.

[0072] In some embodiments, a pharmaceutical composition of the present disclosure includes a therapeutically effective amount of one or more of any of the compounds of the present disclosure, and a pharmaceutically acceptable carrier.

[0073] The compounds of the present disclosure can be incorporated into a variety of compositions for therapeutic administration. More particularly, a compound of the present disclosure can be formulated into pharmaceutical compositions by combination with appropriate, pharmaceutically acceptable excipients or diluents, and may be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, injections, inhalants, and aerosols.

[0074] The disclosed pharmaceutical compositions for administration to a subject (e.g., a human) are generally sterile and may further be free of detectable pyrogens or other contaminants contraindicated for administration to a patient according to a selected route of administration, including but not limited to, parenteral, inhalational, intranasal, subcutaneous, intramuscular, and / or intravenous administration.

[0075] For oral preparations, the compound can be used alone or in combination with appropriate additives to make tablets, powders, granules or capsules, for example, with conventional additives, such as lactose, mannitol, corn starch or potato starch; with binders, such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatins; with disintegrators, such as corn starch, potato starch or sodium carboxymethylcellulose; with lubricants, such as talc or magnesium stearate; and if desired, with diluents, buffering agents, moistening agents, preservatives and flavoring agents.

[0076] A compound of the present disclosure can be formulated for parenteral (e.g., intravenous, intra-arterial, intraosseous, intramuscular, intracerebral, intracerebroventricular, intrathecal, subcutaneous, etc.) administration. In certain aspects, the compound is formulated for injection by dissolving, suspending or emulsifying the compound in an aqueous or non-aqueous solvent, such as vegetable or other similar oils, synthetic aliphatic acid glycerides, esters of higher aliphatic acids or propylene glycol; and if desired, with conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifying agents, stabilizers and preservatives.

[0077] Pharmaceutical compositions that include a compound of the present disclosure may be prepared by mixing the compound having the desired degree of purity with optional physiologically acceptable carriers, excipients, stabilizers, surfactants, buffers and / or tonicity agents. Acceptable carriers, excipients and / or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such asS-T00387W0001 32917 / 70090 / PCphosphate, citrate, and other organic acids; antioxidants including ascorbic acid, glutathione, cysteine, methionine and citric acid; preservatives (such as ethanol, benzyl alcohol, phenol, m-cresol, p-chlor-m-cresol, methyl or propyl parabens, benzalkonium chloride, or combinations thereof); amino acids such as arginine, glycine, ornithine, lysine, histidine, glutamic acid, aspartic acid, isoleucine, leucine, alanine, phenylalanine, tyrosine, tryptophan, methionine, serine, proline and combinations thereof; monosaccharides, disaccharides and other carbohydrates; low molecular weight (less than about 10 residues) polypeptides; proteins, such as gelatin or serum albumin; chelating agents such as EDTA; sugars such as trehalose, sucrose, lactose, glucose, mannose, maltose, galactose, fructose, sorbose, raffinose, glucosamine, N-methylglucosamine, galactosamine, and neuraminic acid; and / or non-ionic surfactants such as Tween, Brij Pluronics, Triton-X, or polyethylene glycol (PEG).

[0078] The pharmaceutical composition may be in a liquid form, a lyophilized form or a liquid form reconstituted from a lyophilized form, wherein the lyophilized preparation is to be reconstituted with a sterile solution prior to administration. The standard procedure for reconstituting a lyophilized composition is to add back a volume of pure water (typically equivalent to the volume removed during lyophilization); however, solutions comprising antibacterial agents may be used for the production of pharmaceutical compositions for parenteral administration.Methods of Treatment

[0079] In some embodiments, the present disclosure provides methods of treating neurodegenerative diseases or disorders in a subject in need comprising administering a therapeutically effective amount of the disclosed compounds, or pharmaceutically acceptable salt thereof, or the disclosed pharmaceutical compositions.

[0080] In some embodiments, the present disclosure provides methods of treating neurodegenerative diseases or disorders in a subject in need thereof comprising administering a therapeutically effective amount ofa compound of Formula (I) or compound 7(7) or dipyridamole (DPM), or a pharmaceutically acceptable salt thereof. In some cases, the compound of Formula (I) administered is selectedS-T00387W0001 32917 / 70090 / PCDPM-A24,S-T00387W0001 32917 / 70090 / PCOH DPM-A41, and OH DPM-A42.

[0081] In some embodiments, the compound of Formula (I) administered is selected from DPM-A2, DPM-A6, DPM-A10, DPM-A11, DPM-A13, DPM-A17, DPM-A19, DPM-A20, DPM-A21, DPM-A22, DPM-A23, DPM-A24, DPM-A28, DPM-A36, DPM-A35, DPM-A34, DPM-A39, DPM-A40, DPM-A46, DPM-A47, DPM-A41, DPM-A42, DPM-A43, DPM-A44, DPM-A45, DPM-A48, DPM-A49, DPM-A50, DPM-A52, DPM-A54, and DPM-A55.

[0082] In some embodiments, the compound of Formula (I) administered is DPM. In some embodiments, the compound of Formula (I) administered is DPM-A23.

[0083] DPM is an FDA-approved generic antiplatelet medication (blood thinner) and a coronary vasodilator that is often used in combination with aspirin and / or warfarin after heart attacks and strokes. It was introduced in 1959 (Allaham et al., 2022). Its use lowers the risk of subsequent cardiovascular (CV) events. It is orally bioavailable and is dosed to human adults as 75 to 100 mg tablets, four times a day. This inconvenient dosing schedule has been improved by designing an extended-release formulation, 200 mg taken twice per day, cotableted with 25 mg of aspirin, in a product sold under the brand name Aggrenox® (Aggrenox Monograph, 2015).

[0084] DPM has multiple mechanisms of action, including cyclic nucleotide phosphodiesterase (PDE) inhibition and equilibrative nucleoside transporter (ENT1,2) inhibition, the latter effect leading to increases in extracellular adenosine stores (Gresele et al., 2011; Wang et al., 2013; Maurice et al., 2014; Rehan et al., 2019; Bondarev et al., 2022). Twelve PDE inhibitors were tested for neuroprotective effects (data not shown) and no effect was observed, suggesting that the anti-PDE activity of DPM is not relevant to its potential use here in ALS treatment. Three inhibitors of ENT 1,2 also were tested and no effect was found on MT biology.

[0085] Without wishing to be bound to any particular theory, it is believed that the disclosed compounds treat and / or prevent many aspects of the neuropathology that occur in the compromised / dying motor neurons of ALS patients. The disclosed compounds protect the mitochondria (MT), axons, and dendrites from the insults of the disease, and markedly extend the life of affected motor neurons in neurodegenerative diseases and disorders characterized by motor neuron death. As shown in the Examples herein, the compounds of the disclosure can preserve MT health, as indicated by one or more of MT count, MT length, neurite cumulative area, andS-T00387W0001 32917 / 70090 / PCpersistence of motor neuron viability in motor neurons of ALS patients. Because neurons in other neurodegenerative diseases like AD or PD are characterized by the death of specific sets of neurons that are thought to deteriorate in the same way as motor neurons in ALS patients, it is expected that the compounds of the disclosure can preserve MT health in the affected neurons of patients with neurodegenerative diseases and disorders other than ALS that are characterized by the death of neurons.

[0086] The neuropathology is complex, including: (1) deposition of abnormal protein aggregates, (2) axon / dendrite degeneration, (3) glutamate excitotoxicity, (4) increased oxidative stress, (5) neuroinflammation, (6) altered lipid metabolism, (7) synapse loss, (8) MT dysfunction, and (9) cell death (Brown et al., 2017;Hardiman et al., 2017; Gan et al., 2018; Le Gall et al., 2020; Mead et al., 2023). Any cell type in the central nervous system (CNS) could be responsible for the numerous ALS-related phenotypes, but ALS susceptibility genes strongly overlap with glutamatergic neuron RNA expression versus other cell types (van Rheenen et al., 2021), indicating that the epicenter for susceptibility is in neurons.

[0087] Without wishing to be bound to any particular theory, it is believed that MT dysfunction is a far upstream event in a molecular / cellular cascade of neuropathologies that occur in ALS (Figure 2). This suggests there is a common molecular cascade of neuropathology triggered by diverse factors (Figure 2), that include mitochondrial (MT) dysfunction, which occurs early in ALS / FTD and some other neurodegenerative diseases, such as AD. The final endpoint of the cascade of neuropathology in ALS is the degeneration and death of motor neurons (MN, Figure 2) in the brain (upper MN) and spinal cord (lower MN). This hypothesis predicts that drugs that protect from, delay, arrest, or reverse the progressive MT dysfunction in ALS would delay or even spare neurons from downstream molecular / cellular pathologies, offering significant disease-modifying therapeutic value. Many observations support this hypothesis. Abnormal MT pathology has been observed to occur early in ALS (Neel et al., 2023; Gautam et al., 2022; Smith et al., 2019), consistent with the hypothesis that it triggers other types of neuropathology. Functional MT are required to support axon and dendrite function (Smith and Gallo, 2018) - item 2 above. MT regulate intracellular calcium (Ca2+) homeostasis (Markovinovic et al., 2022; Chen et al., 2021), so that loss of Ca2+homeostasis due to impaired MT explains the neuronal toxicity due to intracellular Ca2+overload from excess glutamate release (Datta and Jaiswal, 2021; item 3 above). Damaged MT are the major source of reactive oxygen species (ROS; Butterfield et al., 2019; item 4). MT DNA is a recognized agonist of innate immune responses (West and Shadel, 2017; Heneka et al., 2018; Zhong et al., 2018; Yu et al., 2020). Release of MT DNA upon MT damage accounts for inflammasome activation and uncontrolled inflammation (item 5). MT are hubs for the biosynthesis of fatty acids and other lipids (Nowinski et al., 2020, item 6). MT localize near synapses to meet their energy demands and to provide calcium buffering (Du et al., 2010), so that synapse loss and its consequences (item 7) arise from impaired MT (Rangaraju et al., 2019; Fernandez et al., 2019). MT have been reported to be a target for abnormal protein aggregates that form in fALS, including ALS-SOD1, -C9orf72, and -TDP43 (Benson et al., 2021; Wang et al., 2021; Smith et al., 2019; Mathis et al., 2019; Choi et al., 2019; Wang et al., 2016), explaining how genetic insults can lead to common downstream pathologies (item 1). MT dysfunction arising from aging, genetic susceptibility, and / or environmental factors couldS-T00387W0001 32917 / 70090 / PCdrive the onset of sALS. When MT become defective, a pore known as the MT permeability transition pore opens, releasing cytochrome C and other factors that lead to cell death (Bock et al., 2020; Robichaux et al., 2023; Green, 2022). Thus, MT are the platform for this final neuropathological consequence in ALS (item 9). Thus, the MT cascade hypothesis (Figure 2) explains most, and perhaps all, of the neuropathologies found in ALS. Thus, an effective MT therapeutic is predicted to stave off the cellular neuropathology that occurs in ALS.

[0088] Both fALS and sALS cause three broad MT phenotypes: (1) MT fragmentation, (2) reduced MT number, and (3) reduced MT function (Jiang et al., 2015; Deng et al., 2015; Dafinca et al., 2016; Wang et al., 2016; Carri et al., 2017; Smith et al., 2019; Mehta et al., 2019; Petrozziello et al., 2022). These phenotypes have been observed in human postmortem tissue, mouse models for ALS, and in ALS IPSC-derived neurons. They also are observed in pre-symptomatic mice, consistent with the model that MT defects precipitate other types of neuropathology.

[0089] The pharmaceutical composition may be administered to any of a variety of individuals. In certain aspects, the individual is a "mammal” or "mammalian,” where these terms are used broadly to describe organisms which are within the class mammalia, including the orders carnivore (e.g., dogs and cats), rodentia (e.g., mice, guinea pigs, and rats), and primates (e.g., humans, chimpanzees, and monkeys). In some embodiments, the individual is a human. In certain embodiments, the individual is an animal model (e.g., a mouse model, a primate model, or the like) of a neurodegenerative disease or disorder.

[0090] The compound is administered in a therapeutically effective amount. The term "therapeutically effective amount” is meant as a dosage sufficient to produce a desired result, e.g., an amount sufficient to effect beneficial or desired therapeutic (including preventative) results, such as a reduction in a symptom of a neurodegenerative disease or disorder, as compared to a control. In some embodiments, the therapeutically effective amount is sufficient to slow the progression of, or reduce, one or more symptoms of a neurodegenerative disease or disorder. According to some embodiments, the therapeutically effective amount slows the progression of, or reduces, one or more of such symptoms by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100% or more, as compared to the one or more symptoms in the absence of the administration of the compound.

[0091] As described above, aspects of the present disclosure include methods for treating an individual having or suspected of having a neurodegenerative disease or disorder. The term "treating” is meant at least an amelioration of one or more symptoms associated with the neurodegenerative disease or disorder of the individual, where amelioration is used in a broad sense to refer to at least a reduction in the magnitude of a parameter, e.g., symptom, associated with the neurodegenerative disease or disorder. As such, treatment also includes situations where the neurodegenerative disease or disorder, or at least one or more symptoms associated therewith, are completely or partially inhibited, e.g., prevented from happening, or stopped, e.g., terminated, such that the individual no longer suffers from the neurodegenerative disease or disorder, or at least the symptoms that characterize the neurodegenerative disease or disorder are reduced.S-T00387W0001 32917 / 70090 / PC

[0092] As used herein, "predisposed” refers to a subject that has an increased likelihood of developing a neurodegenerative disease or disorder due to genetic factors, environmental factors, lifestyle factors, or a combination thereof. This predisposition may be indicated by family history, specific genetic mutations, or other risk factors that enhance the probability of onset, even if the individual has not yet shown any symptoms of the disease.

[0093] Suitable neurodegenerative diseases and disorders to be treated by the disclosed compounds and pharmaceutical compositions include diseases and disorders characterized by MT dysfunction. In some embodiments, the neurodegenerative disease or disorder is a neurodegenerative disease with MT dysfunction. In some embodiments, the neurodegenerative disease or disorder is other neurological diseases with MT dysfunction. In some embodiments, the neurodegenerative disease or disorder is a psychiatric condition with MT dysfunction. In some embodiments, the neurodegenerative disease or disorder is a disease of the eye with MT dysfunction. In some embodiments, the neurodegenerative disease or disorder is other diseases with MT dysfunction.

[0094] In some embodiments, the neurodegenerative disease or disorder is selected from I. Parkinson's disease (PD) and related disorders, including subsets of PD such as Parkinson's disease dementia (PDD), as well as autosomal recessive PARK2 and PARK6-linked Parkinsonism, atypical Parkinsonian syndromes including progressive supranuclear palsy, corticobasal degeneration syndrome, Lewy body dementia (LBD), multiple system atrophy, Guadeloupean Parkinsonism and Lytigo-bodig disease; ii. motor neuron diseases including but not limited to amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), progressive bulbar palsy, pseudobulbar palsy, primary lateral sclerosis, progressive muscular atrophy, spinal muscular atrophy and post-polio syndrome; ill. neuro-inflammatory diseases including but not limited to autoimmune encephalitis, optic neuritis, transverse myelitis, neurosarcoidosis, neuromyelitis optica, traumatic brain injury, post concussive syndrome, multiple sclerosis, chronic traumatic encephalopathy, spinal cord injury; iv.Alzheimer's disease (AD) and related disorders, including but not limited to early stage of an Alzheimer's disorder, mild stage of an Alzheimer's disorder, moderate stage of an Alzheimer's disorder, mild to moderate stage of an Alzheimer's disorder, advanced stage of an Alzheimer's disorder, mild cognitive impairment, vascular dementia, mixed dementia, Pick's disease, argyrophilic grain disease, posterior cortical atrophy, Wernicke-Korsakoff Syndrome; v. prion diseases including but not limited to Creutzfeldt-Jakob disease, Kuru, variant Creutzfeldt-Jakob disease, Gerstmann-Straussler-Scheinker syndrome, fatal familial insomnia; vi. lysosomal storage diseases including but not limited to Fabry disease, mucopolysaccharidoses, Niemann-Pick disease, Tay-Sachs disease; vii. Leukodystrophies including but not limited to Alexander disease, Canavan disease, Krabbe disease, Adrenoleukodystrophy, Refsum disease; viii. Huntington's disease; ix. multiple sclerosis (MS) including but not limited to secondary progressive MS, progressive relapsing MS, benign MS, primary progressive MS; x. Down syndrome; xi. spinal and bulbar muscular atrophy (SMA) including but not limited to acute infantile SMA, chronic juvenile SMA, adult onset SMA; xii. HIV-Associated neurocognitive disorders including but not limited to HIV-associated dementia, mild neurocognitive disorder; xiii.Tourette Syndrome; xiv.S-T00387W0001 32917 / 70090 / PCspinocerebellar ataxia (SCA) including but not limited to dominant SCA1-35, autosomal recessive SCA, X-linked SCA; xv. Dentatorubral pallidoluysian atrophy; xvi. myotonic dystrophy; xvii, schizophrenia and schizoaffective disorder; xviii. Bipolar spectrum disorders; xix. autism and autism spectrum disorders; xx. attention-deficit hyperactivity spectrum disorders; xxi. chronic pain; xxii. alcohol-induced dementia; xxiii. progressive non-fluent aphasia; xxiv. semantic dementia; xxv. spastic paraplegia; xxvi. fibromyalgia; xxvii. post-Lyme disease; xxviii. neuropathies; xxix. withdrawal symptoms; xxx. Alpers' disease; xxxi. cerebro-oculo-facio-skeletal syndrome; xxxii. Wilson's disease; xxxiii. Cockayne syndrome; xxxiv. Leigh's disease and other primary mitochondrial diseases; xxxv. neurodegeneration with brain iron accumulation; xxxvi. opsoclonus myoclonus syndrome; xxxvii. alpha-methylacyl-CoA racemase deficiency; xxxviii. Andermann syndrome; xxxix. Arts syndrome; xxxx.Marinesco-Sjogren syndrome; xxxxi. mitochondrial membrane protein-associated neurodegeneration; xxxxii. pantothenate kinase-associated neurodegeneration; xxxxiii. polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy; xxxxiv. riboflavin transporter deficiency neuronopathy; xxxxv. ataxia telangiectasial; xxxxvi. age associated memory impairment; and xxxxvii Batten disease.

[0095] In some embodiments, the neurodegenerative disease or disorder is selected from Amyotrophic Lateral Sclerosis (ALS), Alzheimer's disease (AD), Lewy Body dementia (LBD), Parkinson's disease (PD), Huntington's disease, spinal muscular atrophy (SMA), primary lateral sclerosis (PLS), progressive muscular atrophy (PMA), Kennedy's disease (Spinal and Bulbar Muscular Atrophy), post-polio syndrome, motor neuron disease (MND), multiple sclerosis (MS), multifocal motor neuropathy (MMN), hereditary spastic paraplegia (HSP), myasthenia gravis, and Friedreich's ataxia.

[0096] In some embodiments, the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), or Parkinson's disease (PD).Efficacy Across Multiple Genetic Forms of ALS

[0097] The disclosed compounds exhibit efficacy in offsetting five cellular pathologies observed in ALS: mitochondrial fragmentation, loss of mitochondrial content, loss of mitochondrial bioenergetics, the degeneration of axons and dendrites, and early motor neuron death. Importantly, the disclosed compounds demonstrate efficacy across multiple distinct genetic forms of ALS, including for example, C9orf72-ALS, SOD1-ALS, and TDP43-ALS.

[0098] The three genetic forms of ALS - C9orf72, SOD1, and TDP43- represent distinct causes for ALS. The gene C9orf72 is thought to encode a protein involved in lysosomal function. The SOD1 gene encodes a protein that decreases oxidative stress. The TDP43 gene encodes a protein that binds to DNA and RNA and alters gene and protein expression. Despite the distinct insults due to mutations in these three different genes, the general molecular / cellular and organismal pathology observed in the ALS patients is similar. The data presented herein demonstrates that the disclosed molecules work across distinct genetic forms of ALS - rather than being specific to one genetic form - and strongly suggest that the molecules have efficacy in sporadic ALS as well.S-T00387W0001 32917 / 70090 / PC

[0099] In some embodiments, the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS) associated with a mutation in the C9orf72 gene. In some embodiments, the C9orf72 mutation is hexanucleotide repeat expansion. In some embodiments, the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS) associated with a mutation in the TDP43 gene. In some embodiments, the TDP43 mutation is M337V. In some embodiments, the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS) associated with a mutation in the SOD1 gene. In some embodiments, the SOD1 mutation is A5V. In some embodiments, the neurodegenerative disease or disorder is sporadic ALS.Efficacy in C9orf72-ALS Motor Neurons

[0100] Figures 4A-C illustrate data measuring MT count, MT length, and axon / dendrite integrity (Neurite CA) across days 8-30 of culture for ALS-C9orf72 motor neurons (MN) carrying a hexanucleotide repeat expansion mutation and the isogenic control MN. Compounds of the disclosure improve all three parameters in the C9orf72 mutant MN relative to untreated (DMSO) C9orf72 mutant MN, so that the measured values are increased to levels similar to the control MN, as further described in the Examples. Dose:response data (Figure 10A-C) show that the potency of the compound (DPM) is in the low micromolar range. These data demonstrate that the compounds (DPM) offset the loss of MT content, MT fragmentation, and axon / dendrite degeneration due to the disease caused by the C9orf72 mutation.

[0101] In some embodiments, the disclosed compounds, when administered to C9orf72-ALS motor neurons, increase MT count to levels similar to control motor neurons. In some embodiments, the disclosed compounds, when administered to C9orf72-ALS motor neurons, increase MT length to levels similar to control motor neurons. In some embodiments, the disclosed compounds, when administered to C9orf72-ALS motor neurons, increase neurite cumulative area to levels similar to control motor neurons. In some embodiments, the EC50 for MT count in C9orf72 motor neurons is about 0.7-1.2 μM. In some embodiments, the EC50 for MT length in C9orf72 motor neurons is about 1.5-1.6 μM. In some embodiments, the EC50 for neurite cumulative area in C9orf72 motor neurons is about 0.4-0.7 μM.

[0102] Figures 7A-7D illustrate that compounds of the disclosure rescue the deficit in MT bioenergetics observed in the C9orf72 iPSC-derived neurons. In some embodiments, the potency (EC50) of the disclosed compounds in C9orf72 iPSC-derived neurons is between about 0.04 μM and about 0.075 μM. In some embodiments, the EC50 is about 0.04 μM. In some embodiments, the EC50 is about 0.068 μM. In some embodiments, the EC50 is about 0.075 μM.Efficacy in TDP43-ALS Motor Neurons

[0103] Figures 13A-13C illustrate data measuring MT count, MT length, and axon / dendrite integrity (Neurite CA) across days 12-30 of culture for ALS-TDP43 motor neurons (MN) carrying a M337V mutation and the control MN. Compounds of the disclosure improve all three parameters in the TDP43 mutant MN relative to untreated (DMSO) TDP43 mutant MN, so that the measured values are increased to levels similar to the control MN, as further described in the Examples. Dose:response data (bottom) show that the potency of the compound (DPM)S-T00387W0001 32917 / 70090 / PCis in the low micromolar range. These data demonstrate that the compounds offset the loss of MT content, MT fragmentation, and axon / dendrite degeneration due to the disease caused by the TDP43 mutation.

[0104] In some embodiments, the disclosed compounds, when administered to TDP43-ALS motor neurons, increase MT count to levels similar to control motor neurons. In some embodiments, the disclosed compounds, when administered to TDP43-ALS motor neurons, increase MT length to levels similar to control motor neurons. In some embodiments, the disclosed compounds, when administered to TDP43-ALS motor neurons, increase neurite cumulative area to levels similar to control motor neurons. In some embodiments, the EC50 for MT count in TDP43 M337V motor neurons is about 1.5 μM. In some embodiments, the EC50 for MT length in TDP43 M337V motor neurons is about 1.9 μM. In some embodiments, the EC50 for neurite cumulative area in TDP43 M337V motor neurons is about 1.3 μM.Efficacy in SOD1-ALS Motor Neurons

[0105] Figures 14A-14C illustrate data measuring MT count, MT length, and axon / dendrite integrity (Neurite CA) across days 8-30 of culture for ALS-SOD1 motor neurons (MN) carrying an ALS-SOD1 A5V mutation and the control MN. Compounds of the disclosure improved all three parameters in the ALS-SOD1 A5V mutant MN relative to untreated (DMSO) ALS-SOD1 A5V mutant MN, so that the measured values are increased to levels similar to the control MN, as further described in the Examples. Dose:response data (bottom) show that the potency of the compound (DPM) is in the low micromolar range. These data demonstrate that the compounds offset the loss of MT content, MT fragmentation, and axon / dendrite degeneration due to the disease caused by the mutations at the SOD1 gene.

[0106] In some embodiments, the disclosed compounds, when administered to SOD1-ALS motor neurons, increase MT count to levels similar to control motor neurons. In some embodiments, the disclosed compounds, when administered to SOD1-ALS motor neurons, increase MT length to levels similar to control motor neurons. In some embodiments, the disclosed compounds, when administered to SOD1-ALS motor neurons, increase neurite cumulative area to levels similar to control motor neurons. In some embodiments, the EC50 for MT count in SOD1 A5V motor neurons is about 3.7 μM. In some embodiments, the EC50 for MT length in SOD1 A5V motor neurons is about 2.8 μM. In some embodiments, the EC50 for neurite cumulative area in SOD1 A5V motor neurons is about 2.1 μM.Efficacy in Alzheimer's Disease

[0107] Alzheimer's disease (AD) is the most common neurodegenerative disease, affecting millions of individuals worldwide. Like ALS, AD can be characterized by mitochondrial dysfunction, which occurs early in disease progression. There is a need for disease modifying therapeutics for AD. Very few, if any of the FDA approved drugs for AD, have significant efficacy.

[0108] The disclosed compounds also demonstrate efficacy in offsetting pathologies found in Alzheimer's disease (AD) patients and in AD IPSC-derived cortical glutamatergic neurons (herein referred to as corticalS-T00387W0001 32917 / 70090 / PCneurons). The molecules offset five cellular pathologies observed in neurodegenerative disease: mitochondrial fragmentation, loss of mitochondrial content, loss of mitochondrial bioenergetics, the degeneration of axons and dendrites, and early neuronal death, as compared to isogenic control neurons.

[0109] This data is important because it demonstrates that the molecules have similar efficacy in AD iPSC-derived neurons and therefore could have efficacy in patients across the spectrum of neurodegenerative diseases including ALS, AD, Parkinson's disease, and others.

[0110] In some embodiments, the neurodegenerative disease or disorder is Alzheimer's disease (AD). In some embodiments, the Alzheimer's disease is associated with a mutation in the presenilin 1 (PS1) gene. In some embodiments, the Alzheimer's disease is associated with a mutation in the presenilin 2 (PS2) gene. In some embodiments, the Alzheimer's disease is associated with a mutation in the Amyloid Precursor Protein (APP) gene. In some embodiments, the Alzheimer's disease is associated with a mutation in any other gene that influences susceptibility to Alzheimer disease. In some embodiments, the Alzheimer's disease is sporadic Alzheimer's disease.

[0111] Figures 11A-C illustrate data measuring MT count, MT length, and axon / dendrite integrity (Neurite CA) across days 12-30 of culture in AD iPSC-derived cortical neurons. Compounds of the disclosure improved all three parameters relative to untreated AD iPSC-derived neurons, with potency of a representative compound (DPM-A19) between 0.7 and 1.8 μM, as further described in the Examples. These data demonstrate that the compounds offset the loss of MT content, MT fragmentation, and axon / dendrite degeneration due to Alzheimer's disease.

[0112] Figures 12A-12D illustrate that compounds of the disclosure (DPM) rescue the deficit in MT bioenergetics observed in the AD iPSC-derived neurons.

[0113] In some embodiments, the disclosed compounds, when administered to AD iPSC-derived cortical neurons, increase MT count relative to untreated AD neurons. In some embodiments, the disclosed compounds, when administered to AD iPSC-derived cortical neurons, increase MT length relative to untreated AD neurons. In some embodiments, the disclosed compounds, when administered to AD iPSC-derived cortical neurons, increase neurite cumulative area relative to untreated AD neurons. In some embodiments, the disclosed compounds rescue the deficit in MT bioenergetics observed in AD iPSC-derived neurons.

[0114] In some embodiments, the potency (EC50) of the disclosed compounds in AD iPSC-derived neurons is between about 0.7 μM and about 2.2 μM. In some embodiments, the EC50 is about 0.7 μM. In some embodiments, the EC50 is about 1.3 μM. In some embodiments, the EC50 is about 1.4 μM. In some embodiments, the EC50 is about 1.8 μM. In some embodiments, the EC50 is about 2.2 μM.S-T00387W0001 32917 / 70090 / PCEfficacy Across Neuronal Cell Types

[0115] The disclosed compounds demonstrate efficacy in multiple neuronal cell types. The data presented herein demonstrates efficacy in both motor neurons (in the context of ALS) and cortical neurons (in the context of AD).

[0116] In some embodiments, the neuron is a motor neuron. In some embodiments, the neuron is a glutamatergic (cortical) neuron. In some embodiments, the neuron is a cholinergic neuron. In some embodiments, the neuron is a dopaminergic neuron.

[0117] Without wishing to be bound by any particular theory, it is believed that the disclosed compounds protect mitochondria in neurons generally, regardless of the specific neuronal cell type, and therefore have broad applicability across neurodegenerative diseases affecting different neuronal populations.Mechanism of Action: Promotion of Pyruvate Uptake into Mitochondria

[0118] Without wishing to be bound by any particular theory, it is believed that the disclosed compounds work by promoting pyruvate uptake into mitochondria.

[0119] Pyruvate is a key substrate for mitochondrial energy production, entering the mitochondrial matrix via the Mitochondrial Pyruvate Carrier (MPC) where it is converted to Acetyl-CoA and enters the tricarboxylic acid (TCA) cycle. Enhanced pyruvate uptake supports mitochondrial bioenergetics and overall mitochondrial health. See Figure 15.

[0120] In some embodiments, the present disclosure provides methods of promoting pyruvate uptake into mitochondria in a subject in need thereof, comprising administering a therapeutically effective amount of the disclosed compounds, or pharmaceutically acceptable salts thereof, or the disclosed pharmaceutical compositions. In some embodiments, the present disclosure provides methods of treating a disease or disorder characterized by impaired mitochondrial pyruvate uptake, comprising administering a therapeutically effective amount of the disclosed compounds, or pharmaceutically acceptable salts thereof, or the disclosed pharmaceutical compositions. In some embodiments, the compound enhances pyruvate transport across the mitochondrial membrane.

[0121] In some embodiments, the disclosed compounds increase pyruvate uptake into mitochondria in a dose-dependent manner. In some embodiments, the disclosed compounds (e.g., DPM, and DPM-A23) increase pyruvate uptake into mitochondria at concentrations ranging from about 0.1 μM to about 10 μM. In some embodiments, the disclosed compounds increase pyruvate uptake into mitochondria at concentrations of about 0.1 μM, about 0.5 μM, about 1 μM, about 5 μM, or about 10 μM.

[0122] In contrast to known MPC inhibitors such as UK5099, which inhibit pyruvate uptake into mitochondria, the disclosed compounds (e.g., DPM, and DPM-A23) enhance pyruvate uptake. This enhanced pyruvate uptake is believed to support mitochondrial function and contribute to the neuroprotective effects observed in motor neurons and other neuronal cell types. See Figure 16.S-T00387W0001 32917 / 70090 / PCMethods of Preserving Mitochondrial Health

[0123] In some embodiments, the present disclosure provides methods of preserving mitochondrial health in motor neurons of a subject having a neurodegenerative disease, comprising administering a therapeutically effective amount of the disclosed compounds, or pharmaceutically acceptable salts thereof, wherein the method results in one or more of: (a) increased mitochondrial count; (b) increased mitochondrial length; (c) increased neurite cumulative area; (d) rescued mitochondrial bioenergetics; and (e) extended neuronal viability.

[0124] In some embodiments, the present disclosure provides methods of preserving mitochondrial health in motor neurons in subjects predisposed to a neurodegenerative disease.

[0125] In some embodiments, the neurodegenerative disease is selected from ALS and Alzheimer's disease. In some embodiments, the ALS is selected from C9orf72-ALS, SOD1-ALS, TDP43-ALS, and sporadic ALS. In some embodiments, the method results in mitochondrial count, mitochondrial length, and / or neurite cumulative area that is increased to levels similar to those observed in non-diseased control neurons. In some embodiments, the method extends neuronal viability by at least 10 days, at least 20 days, at least 30 days, at least 40 days, at least 50 days, at least 60 days, at least 70 days, or at least 80 days beyond the normal lifespan of untreated diseased neurons.EMBODIMENTS1. A compound of formula (I):or a pharmaceutically acceptable salt thereof,whereinA1is selected from morpholinyl substituted with 0-2 substituents independently selected from Ci-salkyl and Ci-3hydroxyalkyl; piperidinyl substituted with 0-2 substituents independently selected from Ci-salkyl, hydroxy,O O OO 6and Ci-3hydroxyalkyl; -N(R3)2, -J-, ~~L~, and -L-;A2is selected from piperidinyl, pyrrolidinyl, azetidinyl, -N(R4)2, -O-Ci-shydroxyalkyl, -S-Ci-shydroxyalkyl, halo, and morpholinyl substituted with 0-2 substituents independently selected from C^alkyl, and Ci. shydroxyalkyl, wherein the piperidinyl, and pyrrolidinyl are substituted with 0-3 substituents independently selected from C^alkyl, Cvsalkoxy, hydroxy, Ci-shydroxyalkyl, phenyl, and halo;A1or A2optionally further comprise a labile linker;R1and R2are each independently N- or CH, wherein R1and R2together with carbon and nitrogen atoms to which they are attached form a 5- or 6-membered ring, having 0-2 ring N atoms, and the 5- or 6-S-T00387W0001 32917 / 70090 / PCmembered ring is substituted with 0-2 substituents independently selected from A3and -Ci-salkylene-O-Ci-salkyl;each A3is independently selected from morpholinyl substituted with 0-2 substituents independently selected from C^alkyl, and Ci-shydroxyalkyl; piperidinyl, pyrrolidinyl, -N(R5)2, -Ci-shydroxyalkyl, and -Ci.3alkylene-Ci-3alkoxy, wherein the piperidinyl and pyrrolidinyl are substituted with 0-2 substituents independently selected from C^alkyl, hydroxy, and Ci-shydroxyalkyl;each R3is independently Ci-ealkyl or Ci-shydroxyalkyl; andeach R4and R5is independently selected from H, Ci-ealkyl, Ci-ehydroxyalkyl, and Ci-salkyl-O-Ci.ealkylene; with the proviso that the compound is not, and at least one of A1, A2, and A3is piperidinyl or pyrrolidinyl optionally substituted with 0-3 substituents independently selected from Ci-salkyl, C salkoxy, hydroxy, Ci-shydroxyalkyl, phenyl, and halo, or at least one of A1, A2, and A3is morpholinyl optionally substituted with 0-2 substituents independently selected from Ci-salkyl and Ci-shydroxyalkyl, or at least one R4or R5is Ci-ehydroxyalkyl, and Ci.3alkyl-O-Ci-6alkylene.2. The compound or pharmaceutically acceptable salt of embodiment 1, wherein A1is selected3. The compound or pharmaceutically acceptable salt of embodiment 1 or 2, wherein A2isS-T00387W0001 32917 / 70090 / PC4. The compound or pharmaceutically acceptable salt of any one of embodiments 1-3, wherein at least one of R1and R2is Ci-salkyl.5. The compound or pharmaceutically acceptable salt of any one of embodiments 1-4, having aX1and X2are each independently N or CH.6. The compound or pharmaceutically acceptable salt of embodiment 5, wherein at least one X1is N.7. The compound or pharmaceutically acceptable salt of embodiment 5, wherein at least one X1is CH.8. The compound or pharmaceutically acceptable salt of any one of embodiments 5-7, having a A1A1formulaA1or A19. The compound or pharmaceutically acceptable salt of any one of embodiments 1-4, having aX2is N or CH; andA4, A5, and A6are each independently H, C1-4alkyl, morpholinyl substituted with 0-2 substituents independently selected from Ci-salkyl, and Ci-shydroxyalkyl, piperidinyl, pyrrolidinyl, -N(R5)2, Ci-shydroxyalkyl, and Ci-3alkylene-O-Ci-3alkyl, wherein the morpholinyl, piperidinyl and pyrrolidinyl are substituted with 0-2 substituents independently selected from C^alkyl, hydroxy, and Ci-shydroxyalkyl.10. The compound or pharmaceutically acceptable salt of embodiment 9, wherein X2is N.11. The compound or pharmaceutically acceptable salt of embodiment 9, wherein X2is CH. 12. The compound or pharmaceutically acceptable salt of any one of embodiments 9-11, wherein A4is H.S-T00387W0001 32917 / 70090 / PC13. The compound or pharmaceutically acceptable salt of any one of embodiments 9-11, wherein A4is Ci-4alkyl.14. The compound or pharmaceutically acceptable salt of any one of embodiments 9-11, wherein A4is selected from morpholinyl substituted with 0-2 substituents independently selected from Ci-salkyl and Ci. shydroxyalkyl, piperidinyl, pyrrolidinyl, -N(R5)2, -Ci-shydroxyalkyl, and -Ci-salkylhydroxy-Ci-salkyl, wherein the morpholinyl, piperidinyl and pyrrolidinyl are substituted with 0-2 substituents independently selected from Ci¬ salkyl, hydroxy, and Ci-shydroxyalkyl.15. The compound or pharmaceutically acceptable salt of embodiment 14, wherein A4is selected OHfrom H, —I™, and16. The compound or pharmaceutically acceptable salt of any one of embodiments 9-15, wherein A5is H.17. The compound or pharmaceutically acceptable salt of any one of embodiments 9-15, wherein A5is Ci-4alkyl.18. The compound or pharmaceutically acceptable salt of any one of embodiments 9-17, wherein A6is H.19. The compound or pharmaceutically acceptable salt of any one of embodiments 9-17, wherein A6is Ci-4alkyl.20. The compound or pharmaceutically acceptable salt of any one of embodiments 1-19, wherein the labile linker comprises a functional group selected from a hydrazone, a disulfide, an ester, a carbamate, a peptide, an azo, and an oxime.21. The compound or pharmaceutically acceptable salt of any one of embodiments 1-20, wherein the labile linker comprises a phosphate or phosphonate ester, sulfate or sulfonic ester, or a carboxylic ester, wherein the ester comprises an alkyl or amino acid residue.22. The compound or pharmaceutically acceptable salt of embodiment 1, selected fromS-T00387W0001 32917 / 70090 / PCS-T00387W0001 32917 / 70090 / PC DPM-A20,S-T00387W0001 32917 / 70090 / PCS-T00387W0001 32917 / 70090 / PCDPM-A61.23. The compound or pharmaceutically acceptable salt of embodiment 22, selected from 1,S-T00387W0001 32917 / 70090 / PCS-T00387W0001 32917 / 70090 / PCA40, OH DPM-A41, and OH DPM-A42.24. The compound or pharmaceutically acceptable salt of embodiment 22, selected from DPM-A2, DPM-A6, DPM-A10, DPM-A11, DPM-A13, DPM-A17, DPM-A19, DPM-A20, DPM-A21, DPM-A22, DPM-A23, DPM-A24, DPM-A28, DPM-A36, DPM-A35, DPM-A34, DPM-A39, DPM-A40, DPM-A46, DPM-A47, DPM-A41, DPM-A42, DPM-A43, DPM-A44, DPM-A45, DPM-A48, DPM-A49, DPM-A50, DPM-A52, DPM-A54, and DPM- A55.25. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the compound or pharmaceutically acceptable salt of any one of embodiments 1-24.26. The pharmaceutical composition of embodiment 25, wherein the pharmaceutical composition is formulated for oral, parenteral, inhalational, intranasal, subcutaneous, intramuscular, or intravenous administration.27. The pharmaceutical composition of embodiment 26, wherein the pharmaceutical composition is formulated for oral administration.28. The pharmaceutical composition of any one of embodiments 25-27, further comprising an additional therapeutic agent.29. A method of treating a neurodegenerative disease or disorder in a subject diagnosed or predisposed to: i. Parkinson's disease and related disorders including but not limited to Parkinson's disease, Parkinson-dementia, autosomal recessive PARK2 and PARK6-linked Parkinsonism, atypical parkinsonianS-T00387W0001 32917 / 70090 / PCsyndromes, including, progressive supranuclear palsy, corticobasal degeneration syndrome, Lewy bodies dementia, multiple system atrophy, Guadeloupean Parkinsonism and Lytigo-bodig disease; ii. motor neuron diseases including but not limited to amyotrophic lateral sclerosis, frontotemporal dementia, progressive bulbar palsy, pseudobulbar palsy, primary lateral sclerosis, progressive muscular atrophy, spinal muscular atrophy, flail arm syndrome, flail leg syndrome, ALS-plus syndrome, and post-polio syndrome; iii. neuro-inflammatory diseases including but not limited to autoimmune encephalitis, optic neuritis, transverse myelitis, neurosarcoidosis, neuromyelitis optica, traumatic brain injury, post concussive syndrome, multiple sclerosis, chronic traumatic encephalopathy, spinal cord injury; iv. Alzheimer's disease and related disorders including but not limited to early stage of an Alzheimer's disorder, mild stage of an Alzheimer's disorder, moderate stage of an Alzheimer's disorder, mild to moderate stage of an Alzheimer's disorder, advanced stage of an Alzheimer's disorder, mild cognitive impairment, vascular dementia, mixed dementia, Pick's disease, argyrophilic grain disease, posterior cortical atrophy, Wernicke-Korsakoff Syndrome; v. prion diseases including but not limited to Creutzfeldt-Jakob disease, Kuru, variant Creutzfeldt-Jakob disease, Gerstmann-Straussler-Scheinker Syndrome, fatal familial insomnia; vi. lysosomal storage diseases including but not limited to Fabry disease, mucopolysaccharidoses, Niemann-Pick disease, Tay-Sachs disease; vii. Leukodystrophies including but not limited to Alexander disease, Canavan disease, Krabbe disease, Adrenoleukodystrophy, Refsum disease; viii. Huntington's disease; lx. multiple sclerosis (MS) including but not limited to secondary progressive MS, progressive relapsing MS, benign MS, primary progressive MS; x. Down syndrome; xi. spinal and bulbar muscular atrophy (SMA) including but not limited to acute infantile SMA, chronic juvenile SMA, adult onset SMA; xii. HIV-Associated Neurocognitive Disorders including but not limited to HIV-associated dementia, mild neurocognitive disorder; xiii.Tourette Syndrome; xiv. spinocerebellar ataxia (SCA) including but not limited to dominant SCA1-35, autosomal recessive SCA, X-linked SCA; xv. Dentatorubral pallidoluysian atrophy; xvi. myotonic dystrophy; xvii, schizophrenia and schizoaffective disorder; xviii. Bipolar spectrum disorders; xix. autism and autism spectrum disorders; xx. attention-deficit hyperactivity spectrum disorders; xxi. chronic pain; xxii. alcohol-induced dementia; xxiii. progressive non-fluent aphasia; xxiv. semantic dementia; xxv. spastic paraplegia; xxvi. fibromyalgia; xxvii. post-Lyme disease; xxviii. neuropathies; xxix. withdrawal symptoms; xxx. Alpers' disease; xxxi. cerebro-oculo-facio-skeletal syndrome; xxxii. Wilson's disease; xxxiii. Cockayne syndrome; xxxiv. Leigh's disease and other primary mitochondrial diseases; xxxv. neurodegeneration with brain iron accumulation; xxxvi. opsoclonus myoclonus syndrome; xxxvii. alpha-methylacyl-CoA racemase deficiency; xxxviii. Andermann syndrome; xxxix. Arts syndrome; xxxx. Marinesco-Sjogren syndrome; xxxxi. mitochondrial membrane protein-associated neurodegeneration; xxxxii. pantothenate kinase-associated neurodegeneration; xxxxiii. polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy; xxxxiv. riboflavin transporter deficiency neuronopathy; xxxxv. ataxia telangiectasial; xxxxvi. age associated memory impairment; and Batten disease by administering a therapeutically effective amount of the compound or pharmaceutically acceptable salt of any one of embodiments 1-24 or the pharmaceutical composition of any one of embodiments 25-28.S-T00387W0001 32917 / 70090 / PC30. The method of embodiment 29, wherein the neurodegenerative disease or disorder is selected from Amyotrophic Lateral Sclerosis (ALS), Alzheimer's disease (AD), Lewy body dementia (LBD), Parkinson's disease (PD), Huntington's disease, spinal muscular atrophy (SMA), primary lateral sclerosis (PLS), progressive muscular atrophy (PMA), Kennedy's disease (spinal and bulbar muscular atrophy), post-polio syndrome, motor neuron disease (MND), multiple sclerosis (MS), multifocal motor neuropathy (MMN), hereditary spastic paraplegia (HSP), myasthenia gravis, and Friedreich's ataxia.31. The method of embodiment 30, wherein the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), or Parkinson's disease (PD).32. The method of any one of embodiments 29-31, wherein the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), progressive muscular atrophy, primary lateral sclerosis, progressive bulbar palsy, flail arm syndrome, flail leg syndrome, and ALS-plus syndrome.33. The compound or pharmaceutically acceptable salt of any one of embodiments 1-24 or the pharmaceutical composition of any one of embodiments 25-28, for use in treating a neurodegenerative disease or disorder.34. The method of embodiment 29, wherein the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS) associated with a mutation in the C9orf72 gene.35. The method of embodiment 34, wherein the C9orf72 mutation is a hexanucleotide repeat expansion.36. The method of embodiment 29, wherein the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS) associated with a mutation in the TDP43 gene.37. The method of embodiment 36, wherein the TDP43 mutation is M337V.38. The method of embodiment 29, wherein the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS) associated with a mutation in the SOD1 gene.39. The method of embodiment 38, wherein the SOD1 mutation is A5V.40. The method of embodiment 29, wherein the neurodegenerative disease or disorder is sporadic ALS.41. The method of embodiment 29, wherein the neurodegenerative disease or disorder is Alzheimer's disease (AD).42. The method of embodiment 41, wherein the Alzheimer's disease is associated with a mutation in the presenilin 1 (PS1) gene or presenilin 2 (PS2) gene.43. The method of embodiment 41, wherein the Alzheimer's disease is sporadic Alzheimer's disease.S-T00387W0001 32917 / 70090 / PC44. A method of promoting pyruvate uptake into mitochondria in a subject in need thereof, comprising administering a therapeutically effective amount of the compound or pharmaceutically acceptable salt of any one of embodiments 1-24 or the pharmaceutical composition of any one of embodiments 25-28.45. The method of embodiment 44, wherein the compound enhances pyruvate transport across the mitochondrial membrane.46. The method of embodiment 44 or 45, wherein pyruvate transport enhancement is mediated by Mitochondrial Pyruvate Carrier (MPC).47. A method of treating a disease or disorder characterized by impaired mitochondrial pyruvate uptake, comprising administering a therapeutically effective amount of the compound or pharmaceutically acceptable salt of any one of embodiments 1-24 or the pharmaceutical composition of any one of embodiments 25-28.48. A method of preserving mitochondrial health in motor neurons of a subject having a neurodegenerative disease, comprising administering a therapeutically effective amount of the compound or pharmaceutically acceptable salt of any one of embodiments 1-24, wherein the method results in one or more of: (a) increased mitochondrial count; (b) increased mitochondrial length; (c) increase mitochondrial bioenergetics; (d) increased neurite cumulative area; and (e) extended motor neuron viability.49. The method of embodiment 47 or 48, wherein the neurodegenerative disease is selected from ALS and Alzheimer's disease.50. The method of embodiment 47 or 48, wherein the neurons are selected from motor neurons and cortical neurons.51. The method of embodiment 47 or 48, wherein the ALS is selected from C9orf72-ALS, SOD1-ALS, TDP43-ALS, other fALS forms, and sporadic ALS.52. The method of embodiment 47 or 48, wherein the Alzheimer's disease is associated with a mutation in the presenilin 1 (PS1) gene or presenilin 2 (PS2) gene, APP gene, other fAD forms, and sporadic AD.53. The method of any one of embodiments 29-52, wherein the compound increases at least one parameter selected from mitochondrial count, mitochondrial length, mitochondrial bioenergetics, neurite cumulative area, and neuronal survival to levels similar to those observed in non-diseased control neurons.54. The method of embodiment 53, wherein the compound increases mitochondrial count, mitochondrial length, and neurite cumulative area.55. The method of any one of embodiments 29-54, wherein the EC50for one or more of mitochondrial count, mitochondrial length, and neurite cumulative area is in the low micromolar range.56. The method of embodiment 55, wherein the EC50 is from about 0.4 μM to about 4 μM.57. The method of embodiment 55, wherein the EC50 is from about 0.7 μM to about 4 μM.S-T00387W0001 32917 / 70090 / PC58. The method of embodiment 53, wherein the compound increases mitochondrial bioenergetics.59. The method of any one of embodiments 29-58, wherein the EC50for mitochondrial bioenergetics is about 0.04 μM to about 2.2 μM.60. The method of any one of embodiments 29-59, wherein the compound is dipyridamole (DPM) or a structural analog thereof.61. The method of embodiment 60, wherein the compound is dipyridamole (DPM).62. The method of embodiment 54, wherein the compound is compound 23 (DPM-A23) of formulaN

[0126] The following Examples are provided to further illustrate aspects of the disclosure and are not meant to constrain the disclosure to any particular application or theory of operation.

[0127] All publications, patents and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference.EXAMPLES

[0128] The following examples further illustrate the disclosed methods of treatment, but of course, should not be construed as in any way limiting its scope.COMPOUND SYNTHESIS

[0129] General Procedures. Starting materials were purchased from commercial vendors and used without purification. All moisture-sensitive reactions were performed under argon pressure. Experiments were monitored by LCMS or TLC and visualized using an ultraviolet lamp (254 nm) or staining with KMnO4. Purification via silica gel flash column chromatography was performed using a Teledyne ISCO Combiflash® Rf+, Biotage® Isolera one, or Biotage® Isolera four instrument, with Luknova silica gel cartridges or Biotage® Sfar silica (60 μm). Reverse phase chromatography used Biotage® Sfar C18D (Duo 100 Å, 30 μm) cartridges, with eluents w acetonitrile and water, each with 0.1% TFA or 0.1% formic added. All NMR data was collected at room temperature on a Brüker Ultrashield 400 MHz nuclear magnetic resonance spectrometer. Chemical shifts for1H NMR spectra are reported in parts per million (ppm) relative to residual solvent signal as an internal standard: DMSO (δ 2.50), CHCl3(δ 7.26), Acetone (δ 2.05), or MeOH (δ 3.31). Multiplicities are given as s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), or br (broad peak). Coupling constants are reported as a J value in Hertz (Hz). Mass spectra were recorded on a Thermo Scientific 3000 LCQ Fleet system (ESI) using a Discovery® HS C18 HPLC column (10 cm x 2.1 mm, 5 pm) at 35 °C with UV detection at 254 nm. Flow rate was 0.7 mL / minS-T00387W0001 32917 / 70090 / PCusing a solvent gradient of 5-95% B over 4 min (total run time = 6 min), where A = 0.1 % formic acid in de-ionized H2O and B = 0.1% formic acid in ACN. Reverse phase high-performance preparatory liquid chromatography was conducted using an Agilent 1290 / 1260 Infinity II HPLC system consisting of a G7114B VWD detector, a G7157A prep autosampler, two G7161A prep binary pumps, and a G7159B fraction collector. Data analysis was performed using Agilent's ChemStation® software using an Agilent 10 Prep-C18250 x 30.0 mm column with UV detection at 254 and / or 280 nm. The flow rate was 30 mL / min using a gradient of 10-80% B / A (A = 0.1% TFA in DI H2O and B = 1:1 ACN / MeOH) over 10 min for the necessary length of time to collect pure sample. Microwave-assisted reactions were performed in the Biotage Initiator 4.1.2.

[0130] Scheme 1. Synthetic route for the preparation of synthetic intermediates 2,4,6-Trichloro-8-(piperidin-1-yl)pyrimido[5,4-d]pyrimidine (1) and 2,6-dichloro-4,8-di(piperidin-1 -yl)pyrimido[5,4-c / ]pyrimidine (2).2,4,6-Trichloro-8-(piperidin-1 -yl)pyrimido[5,4-d]pyrimidine (1). A mixture of perchloropyrimido[5,4-d]pyrimidine (1.0 g, 3.7 mmol) and potassium carbonate (1.0 g, 7.0 mmol) in THF was kept at -78°C. Piperidine (0.3 g, 4 mmol) was then added dropwise in THF at a rate of 1.0 mL per minute. The mixture was stirred for 1 hour after the addition. Excess water was added afterward, resulting in the formation of a yellow precipitate. The precipitate was then extracted with ethyl acetate. Upon drying, a yellow glass was obtained. The glass was triturated with diethyl ether, filtered, and dried as a yellow powder, yielding a solid residue 1 in quantitative yield.

[0131] 2,6-Dichloro-4,8-di (piperidin-1 -yl)pyrimido[5,4-d]pyrimidine (2). A mixture of 2,4,6-trichloro-8-(piperidin-1-yl)pyrimido[5,4-d]pyrimidine (1) (500.0 mg, 1.57 mmol) and potassium carbonate (434.0 mg, 3.14 mmol) in THF was kept at 0°C. Piperidine (147.0 mg, 1.73 mmol) was then added dropwise in THF at a rate of 1.0 mL per minute. The mixture was stirred for 1 hour after the addition. Excess water was added afterward, resulting in the formation of a yellow precipitate. The precipitate was then extracted with ethyl acetate. Upon drying, a yellow glass was obtained. The glass was triturated with diethyl ether, filtered, and dried as a yellow powder, yielding a solid residue 2 in quantitative yield. MS (m / z): [M+H] calc'd for C16H21Cl2N6is 367.11, found 367.37.

[0132] 2,2',2",2"'-((4,8-Di(piperidin-1-yl)pyrimido[5,4-d]pyrimidine-2,6-diyl)bis(azanetriyl))tetrakis(ethan-1-ol) (DPM-A3, internal resynthesis of dipyridamole):S-T00387W0001 32917 / 70090 / PC

[0133] A mixture of 2 (30.0 mg, 0.14 mmol), diethanolamine (0.1 mL, 1.4 mmol), and N,N-diisopropylethylamine (47.0 pL, 0.27 mmol) in 1 -butanol (1.0 mL) was stirred in the microwave at 160 °C for 3 hours. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A3 (35.0 mg, 51% yield). MS(m / z): [M+H] calc'd for C24H40N8O4is 505.32, found 505.18.

[0134] 2, 2', 2", 2"', 2"", 2 -((8-(Piperidin-1 -yl)pyrimido[5,4-c / ]pyrimidine-2,4,6-triyl)tris(azanetriyl))hexakis(ethan- 1-ol) (DPM-A2):

[0135] A mixture of 1 (30.0 mg, 0.14 mmol), excess diethanolamine (0.1 mL, 1.4 mmol), and N,N-diisopropylethylamine (33.0 μL, 0.19 mmol) in 1 -butanol (1.0 mL) was stirred in the microwave at 160 °C for 5 hours. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A2 (27.0 mg, 55% yield). MS(m / z): [M+H] calc'd for C24H40N8O4is 525.31, found 525.91.

[0136] 4,4',4"-(8-(Piperidin-1-yl)pyrimido[5,4-c / ]pyrimidine-2,4,6-triyl)trimorpholine (DPM-A5):S-T00387W0001 32917 / 70090 / PC

[0137] A mixture of 1 (30.0 mg, 0.14 mmol), excess morpholine (0.12 mL, 1.4 mmol), and N,N-diisopropylethylamine (33.0 μL, 0.19 mmol) in 1 -butanol (1.0 mL) was stirred in the microwave at 160 °C for 3 hours. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A5 (24.0 mg, 54% yield). MS(m / z): [M+H] calc’d for C23H35N8O3is 471.28, found 471.89.

[0138] Scheme 2. Synthetic route for the preparation of synthetic intermediate 2,6-dichloro-N4,N4,N8,N8-tetraethylpyrimido[5,4-d]pyrimidine-4,8-diamine (4).

[0139] Compounds 3 and 4 were synthesized following the same procedure as used for preparing compounds 1 and 2 with minor modifications. Perchloropyrimido[5,4-d]pyrimidine (1.0 g, 4.0 mmol) and potassium carbonate (1.0 g, 7 mmol) in THF was kept at -78°C. Diethylamine (0.3 g, 4 mmol) was then added dropwise in THF at a rate of 1 mL per minute. The mixture was stirred for 1 hour after the addition. Excess water was added afterward, resulting in the formation of a yellow precipitate. This precipitate was then extracted with ethyl acetate. Upon drying, a yellow glass was obtained. The glass was triturated with diethyl ether, filtered, and dried as a yellow powder, yielding a solid residue 3 in quantitative yield. To a mixture of 3 (500.0 mg, 1.63 mmol) in 10 mL of THF, diethylamine (0.1 mL) was added, and the reaction mixture was stirred at room temperature for 3 hours.Compound 4 was obtained in quantitative yield and purified using a similar work-up procedure as compound 2.MS(m / z): [M+H] calc’d for C14H20Cl2N6is 343.11, found 343.16.

[0140] N,N-diethyl-2,6,8-trimorpholinopyrimido[5,4-d]pyrimidin-4-amine (DPM-A9):

[0141] A mixture of 3 (30.0 mg, 0.098 mmol), excess morpholine (0.13 mL, 1.5 mmol), and N,N-diisopropylethylamine (34.0 μL, 0.20 mmol) in 1,4-dioxane (1.0 mL) was stirred in a sealed vial at 150 °C overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A9 (21.0 mg, 47% yield). MS(m / z): [M+H] calc’d for C22H34N8O3is 459.28, found 459.80.S-T00387W0001 32917 / 70090 / PC

[0142] 2, 2', 2", 2"', 2"", 2 -((8-(Diethylamino)pyrimido[5,4-^pyrimidine-2,4,6-triyl)tris(azanetriyl))hexakis(ethan- 1-ol) (DPM-A8):

[0143] A mixture of 3 (30.0 mg, 0.098 mmol), diethanolamine (0.1 mL, 1.2 mmol), and N,N-diisopropylethylamine (28.0 pL, 0.16 mmol) in 1,4-dioxane (1.0 mL) was stirred in a sealed vial at 150 °C overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A8 (21.0 mg, 47% yield). MS(m / z): [M+H] calc'd for C22H40N8O6 is 513.31, found 513.75.

[0144] A / 4. A / 4, N8, N8-tetraethyl-2,6-dimorpholinopyrimido[5,4-c / ]pyrimidine-4,8-c / / am / ne (DPM-A12):

[0145] A mixture of 4 (30.0 mg, 0.087 mmol), excess morpholine (0.11 mL, 1.3 mmol), and N,N-diisopropylethylamine (30.0 pL, 0.17 mmol) in 1,4-dioxane (1.0 mL) was stirred in a sealed vial at 150 °C overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A12 (15.0 mg, 39% yield). MS(m / z): [M+H] calc'd for C22H36N8O2 is 445.30, found 445.90.

[0146] Scheme 3. Synthetic route for the preparation of synthetic intermediates 4-(2,6,8-trichloropyrimido[5,4- d]pyrimidin-4-yl)morpholine (5) and 4,4'-(2,6-dichloropyrimido[5,4-c / ]pyrimidine-4,8-diyl)dimorpholine (6).S-T00387W0001 32917 / 70090 / PC

[0147] 4-(2,6,8-trichloropyrimido[5,4-c / ]pyrimidin-4-yl)morpholine (5) and compound 6 were synthesized following the same procedure as compounds 1 and 2 with minor modifications. Starting with a mixture of perchloropyrimido[5,4-c / ]pyrimidine (1.0 g, 4.0 mmol) and potassium carbonate (1.0 g, 7.0 mmol) in THF was kept at -78°C. Morpholine (0.4 g, 4.0 mmol) was then added dropwise in THF at a rate of 1 mL per minute. The mixture was stirred for 1 hour after the addition. Excess water was added afterward, resulting in the formation of a yellow precipitate. This precipitate was then extracted with ethyl acetate. Upon drying, a yellow glass was obtained. The glass was triturated with diethyl ether, filtered, and dried as a yellow powder, yielding a solid residue 5 in quantitative yield. To a mixture of 5 (500.0 mg, 1.63 mmol) in 10 mL of THF, diethylamine (0.1 mL) was added, and the reaction mixture was stirred at room temperature for 3 hours. Compound 6 was obtained in quantitative yield and purified using a similar work-up procedure as compound 2. MS(m / z): [M+H] calc'd for C14H16CI2N6O2 is 371.07, found 371.45.

[0148] 2, 2', 2", 2"', 2"", 2 -((8-Morpholinopyrimido[5,4-^pyrimidine-2,4,6-triyl)tris(azanetriyl))hexakis(ethan-1-ol) (DPM-A6):

[0149] A mixture of 5 (30.0 mg, 0.094 mmol), excess diethanolamine (0.10 mL, 1.04 mmol), and N,N-diisopropylethylamine (33.0 μL, 0.19 mmol) in 1 -butanol (1.0 mL) was stirred in the microwave at 160 °C for 3 hours. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A6 (23.0 mg, 47% yield). MS(m / z): [M+H] calc'd for C22H38N8O7 is 527.29, found 526.98.

[0150] 2,2',2",2"'-((4,8-Dimorpholinopyrimido[5,4-c / |pyrimidine-2,6-diyl)bis(azanetriyl))tetrakis(ethan-1-ol) (DPM-A7):

[0151] A mixture of 6 (50.0 mg, 0.13 mmol), diethanolamine (0.1 mL, 1.3 mmol), and N,N-diisopropylethylamine (47.0 μL, 0.27 mmol) in 1 -butanol (1.0 mL) was stirred in the microwave at 160 °C for 3S-T00387W0001 32917 / 70090 / PChours. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A7 (34.0 mg, 50% yield). MS(m / z): [M+H] calc'd for C22H36N8O6 is 509.28, found 509.10.

[0152] DPM-A7b (TFA salt of DPM-A7): DPM-A7 (10.0 mg, 0.02 mmol) was dissolved in 1 mL of methanol. Subsequently, TFA (1.5 pL, 0.02 mmol) was added to the solution and stirred for 15 minutes at room temperature. The solvents were then evaporated under vacuum, and the resulting residue was lyophilized to obtain the product as a TFA salt.

[0153] DPM A7c (HCI salt of DPM-A7): DPM-A7 (10.0 mg, 0.02 mmol) was dissolved in 1 mL of methanol. Subsequently, HCI (4 M in 1-4-dioxane, 4.9 pL, 0.02 mmol) was added to the solution and stirred for 15 minutes at room temperature. The solvents were then evaporated under vacuum, and the resulting residue was lyophilized to obtain the product as a TFA salt.

[0154] 2,2'-((4,8-Dimorpholinopyrimido[5,4-c / ]pyrimidine-2,6-diyl)bis(methylazanediyl))bis(ethan-1-ol) (DPM- A10):

[0155] A mixture of 6 (30.0 mg, 0.081 mmol), excess 2-(methylamino)ethan-1-ol (0.03 mL, 0.4 mmol), and N, N-diisopropylethylamine (70 pL, 0.4 mmol) in 1-4-dioxane (1.0 mL) in the sealed vial was stirred at 160 °C overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A10 (23.0 mg, 63% yield). MS(m / z): [M+H] calc'd for C20H32N8O4 is 449.25, found 448.97.

[0156] 1, T-(4,8-Dimorpholinopyrimido[5,4-c / ]pyrimidine-2,6-diyl)bis(piperidin-3-ol) (DPM-A11):S-T00387W0001 32917 / 70090 / PC

[0157] A mixture of 6 (30.0 mg, 0.081 mmol), racemic piperidin-3-ol (0.04 mL, 0.4 mmol), and N, N-diisopropylethylamine (70.0 pL, 0.4 mmol) in 1-4-dioxane (1.0 mL) in the sealed vial was stirred at 160 °C overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A11 (24.0 mg, 59% yield). MS(m / z): [M+H] calc'd for C24H36N8O4 is 501.29, found 501.06.

[0158] 2,2'-((4,8-Dimorpholinopyrimido[5,4-c / ]pyrimidine-2,6-diyl)bis(azanediyl))bis(ethan-1-ol) (DPM-A13):

[0159] A mixture of 6 (30.0 mg, 0.081 mmol), excess 2-aminoethan-1-ol (0.5 mL, 8.1 mmol), and N, N-diisopropylethylamine (70 pL, 0.4 mmol) was stirred in the microwave at 160 °C for 0.5 h. After completion, the reaction mixture was purified by silica gel chromatography, resulting in the pure product DPM-A13 (15.0 mg, 44% yield). MS(m / z): [M+H] calc’d for C18H28N8O4is 421.22, found 421.00.

[0160] 2,2'-((4,8-Dimorpholinopyrimido[5,4-d]pyrimidine-2,6-diyl)bis(oxy))bis(ethan-1-ol) (DPM-A14):

[0161] A mixture of 6 (30.0 mg, 0.081 mmol), ethane-1,2-diol (0.5 mL, 8.1 mmol), and N, N-diisopropylethylamine (28.0 pL, 0.16 mmol) was stirred in the microwave at 100 °C for 10 minutes. After completion, the reaction mixture was purified by C18 silica gel chromatography, resulting in pure DPM-A14 (12.0 mg, 35% yield). MS(m / z): [M+H] calc’d for C18H26N6O6is 423.19, found 423.53.

[0162] 2,2'-((6-chloro-4,8-dimorpholinopyrimido[5,4-d]pyrimidin-2-yl)azanediyl)bis(ethan-1-ol) (DPM-A15):S-T00387W0001 32917 / 70090 / PC

[0163] A mixture of 6 (50.0 mg, 0.13 mmol), ethane-1,2-diol (0.01 mL, 0.13 mmol), and N, N-diisopropylethylamine (23.0 pL, 0.13 mmol) in 1-4-dioxane ( 2mL) was stirred at 100 °C for overnight. After completion, the reaction mixture was purified by C18 silica gel chromatography, resulting in pure DPM-A15 (43.0 mg, 73% yield). MS(m / z): [M+H] calc’d for C18H26CIN7O4 is 440.17, found 440.15.

[0164] 1,r-(4,8-Dimorpholinopyrimido[5,4-c / ]pyrimidine-2,6-diyl)bis(piperidin-4-ol) (DPM-A17):

[0165] A mixture of 6 (30.0 mg, 0.081 mmol), excess piperidin-4-ol (0.04 mL, 0.4 mmol), and N, N-diisopropylethylamine (70.0 pL, 0.4 mmol) in 1-4-dioxane (1.0 mL) in the sealed vial was stirred at 160 °C overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A17 (18.0 mg, 44% yield). MS(m / z): [M+H] calc’d for C24H36N8O4 is 501.29, found 501.08.

[0166] 2,2'-((4,8-Dimorpholinopyrimido[5,4-d]pyrimidine-2,6-diyl)bis(oxy))bis(ethan-1-ol) (DPM-A18):

[0167] A mixture of 6 (30.0 mg, 0.081 mmol), 2-mercaptoethan-1-ol (0.5 mL), and potassium carbonate (22.0 mg, 0.16 mmol) in N, N-dimethylformamide (1 mL) was irradiated in microwave at 200 °C for 10 minutes. AfterS-T00387W0001 32917 / 70090 / PCcompletion, the reaction mixture was purified by C18 silica gel chromatography, resulting in pure DPM-A18 (15.0 mg, 41% yield). MS(m / z): [M+H] calc’d for C18H26N6O4S2 is 455.15, found 455.70.

[0168] (3R,3'R)-1,r-(4,8-dimorpholinopyrimido[5,4-c / ]pyrimidine-2,6-diyl)bis(pyrrolidin-3-ol) (DPM-A19):

[0169] A mixture of 6 (30.0 mg, 0.081 mmol), excess (R)-pyrrolidin-3-ol (35.0 mg, 0.4 mmol), and N, N-diisopropylethylamine (70.0 pL, 0.4 mmol) in 1-4-dioxane (1.0 mL) in the microwave was stirred at 140 °C for 0.5 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A19 (22.0 mg, 58% yield). MS(m / z): [M+H] calc’d for C22H33N8O4 is 473.25, found 473.90.

[0170] (3S,3'S)-1,1'-(4,8-dimorpholinopyrimido[5,4-c / ]pyrimidine-2,6-diyl)bis(pyrrolidin-3-ol) (DPM-A20):

[0171] A mixture of 6 (30.0 mg, 0.081 mmol), excess (S)-pyrrolidin-3-ol (35.0 mg, 0.4 mmol), and N, N-diisopropylethylamine (70.0 pL, 0.4 mmol) in 1-4-dioxane (1.0 mL) in the microwave was stirred at 140 °C for 0.5 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A20 (23.0 mg, 60% yield). MS(m / z): [M+H] calc’d for C22H32N8O4 is 473.25, found 473.40.

[0172] (3S,3'S)-1,1'-(4,8-dimorpholinopyrimido[5,4-c / ]pyrimidine-2,6-diyl)bis(piperidin-3-ol) (DPM-A21):S-T00387W0001 32917 / 70090 / PC

[0173] A mixture of 6 (30.0 mg, 0.081 mmol), excess (S)-piperidin-3-ol (41.0 mg, 0.4 mmol), and N, N-diisopropylethylamine (70 pL, 0.4 mmol) in 1-4-dioxane (1.0 mL) in the microwave was stirred at 140 °C for 0.5 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A21 (26.0 mg, 64% yield). MS(m / z): [M+H] calc'd for C24H36N8O4 is 501.29, found 501.90.

[0174] (3R,3'R)-1,r-(4,8-dimorpholinopyrimido[5,4-c / ]pyrimidine-2,6-diyl)bis(piperidin-3-ol) (DPM-A22):

[0175] A mixture of 6 (30.0 mg, 0.081 mmol), excess (R)-piperidin-3-ol (41.0 mg, 0.4 mmol), and N, N-diisopropylethylamine (70.0 pL, 0.4 mmol) in 1-4-dioxane (1.0 mL) in the microwave was stirred at 140 °C for 0.5 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A22 (27.0 mg, 67% yield). MS(m / z): [M+H] calc'd for C24H36N8O4 is 501.29, found 501.40.

[0176] N2. N2, N6, N6-tetrakis(2-methoxyethyl)-4,8-dimorpholinopyrimido[5,4-c / ]pyrimidine-2,6-diamine (DPM- A34):S-T00387W0001 32917 / 70090 / PC

[0177] An oven-dried reaction flask was filled with 2,2',2",2"'-((4,8-dimorpholinopyrimido[5,4-d]pyrimidine-2,6-diyl)bis(azanetriyl))tetrakis(ethan-1-ol) (DPM-A7, 30.0 mg, 0.059 mmol) in 1 mL of THF under argon. NaH (7.1 mg, 0.29 mmol) was slowly added at 0°C, and the reaction mixture was stirred for 1 hour. CH3I (0.04 mL, 0.29 mmol) in THF was added dropwise, and the reaction mixture was stirred at room temperature overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by C silica gel flash chromatography, resulting in the pure product DPM-A34 as a TFA salt (20.0 mg, 60% yield). MS(m / z): [M+H] calc'd for C26H44N8O6 is 565.34, found 565.50.

[0178] (3S,3'S,5S,5'S)-1, T-(4,8-dimorpholinopyrimido[5,4-c / ]pyrimidine-2,6-diyl)bis(5-(hydroxymethyl)pyrrolidin-3-ol) (DPM-A35):

[0179] A mixture of 6 (50.0 mg, 0.13 mmol), excess (3S,5S)-5-(hydroxymethyl)pyrrolidin-3-ol hydrochloride (100.0 mg, 0.67 mmol), and N, N-diisopropylethylamine (0.12 mL, 0.4 mmol) in 1 -butanol (1.0 mL) in the microwave was stirred at 180 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A35 (29.0 mg, 40% yield). MS(m / z): [M+H] calc’d for C24H36N8O6 is 533.28, found 533.50.

[0180] (+)1, T-(4,8-Dimorpholinopyrimido[5,4-c / ]pyrimidine-2,6-diyl)bis(pyrrolidin-3-ol) (DPM-A36):

[0181] A mixture of 6 (30.0 mg, 0.081 mmol), excess racemic pyrrolidin-3-ol (35.0 mg, 0.4 mmol), and N, N-diisopropylethylamine (70.0 pL, 0.4 mmol) in 1-4-dioxane (1.0 mL) in the microwave was stirred at 140 °C for 0.5 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A36 (23.0 mg, 60% yield). MS(m / z): [M+H] calc'd for C22H32N8O4 is 473.25, found 473.80.S-T00387W0001 32917 / 70090 / PC

[0182] 1,r-(4,8-dimorpholinopyrimido[5,4-c / ]pyrimidine-2,6-diyl)bis(3-methylpyrrolidin-3-ol) (DPM-A46)

[0183] A mixture of 6 (50.0 mg, 0.13 mmol), racemic 3-methylpyrrolidin-3-ol (68.0 mg, 0.68 mmol), and N, N-diisopropylethylamine (0.12 mL, 0.67 mmol) in 1-butanol (1.0 mL) in the microwave was stirred at 180 °C for 3 hours. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A46 (12.0 mg, 18% yield). MS(m / z): [M+H] calc'd for C24H36N8O4 is 501.29, found 501.60.

[0184] 4,4'-(2,6-bis(3-methoxypyrrolidin-1-yl)pyrimido[5,4-c / ]pyrimidine-4,8-diyl)dimorpholine (DPM-A47)

[0185] A mixture of 6 (50.0 mg, 0.13 mmol), 3-methoxypyrrolidine (68.0 mg, 0.67 mmol), and N,N-diisopropylethylamine (0.12 mL, 0.67 mmol) in 1-butanol (1.0 mL) in the microwave was stirred at 180 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A47 (15.0 mg, 22% yield). MS(m / z): [M+H] calc'd for C24H36N8O4 is 501.29, found 501.60.

[0186] Scheme 4. Synthesis of synthetic intermediates 4,4'-(4,5-Dihydro-9H-purine-2,6-diyl)dimorpholine (7) and (+)-4-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-2-methylmorpholine (22).7: R = H22: R = MeS-T00387W0001 32917 / 70090 / PC

[0187] 4-(2-chloro-4a,7a-dihydro-7H-pyrrolo[2,3-d]pyrimidin-4-yl)morpholine (7) and (+)-4-(2-chloro-7H-py rrolo[2, 3-d] py rimidin-4-y l)-2-methyl morphol ine (22). A mixture of 2,4-dichloro-4a, 7a-dihydro-7 / 7-py rrolo[2,3-c / ]py rimidine (1.0 g, 5.0 mmol) and potassium carbonate (1.0 g, 8.0 mmol) in THF was kept at -78°C. Morpholine (0.48 g, 5.5 mmol) was then added dropwise in THF at a rate of 1 mL per minute. The mixture was stirred for 1 hour after the addition. The reaction mixture was concentrated to dryness and compound 7 was obtained by silica gel chromatography. MS(m / z): [M+H] calc'd for C10H14CIN4O is 241.08, found 241.50. The same procedure was used replacing morpholine with (+)-2-methylmorpholine to give compound 22, MS(m / z): [M+H] calc'd for C11H13ClN4O is 252.70, found 253.30.

[0188] 2,2'-((4-Morpholino-4a,7a-dihydro-7 / - / -pyrrolo[2,3-c / ]pyrimidin-2-yl)azanediyl)bis(ethan-1-ol) (DPM-A23):OH

[0189] A mixture of 7 (100.0 mg, 0.41 mmol), diethanolamine (0.04 mL, 0.41 mmol), and N, N-diisopropylethylamine (145.0 pL, 0.83 mmol) in N-methylpyrrolidone (1.0 mL) in the sealed vial was stirred at 160 °C for 3h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A23 (70.0 mg, 55% yield). MS(m / z): [M+H] calc'd for C14H23N5O3 is 308.16, found 308.30.

[0190] (+)-2,2'-((4-(2-methylmorpholino)-7 / - / -pyrrolo[2,3-c / ]pyrimidin-2-yl)azanediyl)bis(ethan-1-ol) (DPM-A41).OH

[0191] A mixture of 22 (50.0 mg, 0.20 mmol), diethanolamine (0.06 mL, 0.6 mmol), and N, N-diisopropylethylamine (69.0 pL, 0.4 mmol) in 1 -butanol (1.0 mL) in the sealed vial in microwave was stirred at 160 °C for 3h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A41 (34.0 mg, 53% yield). MS(m / z): [M+H] calc’d C15H23N5O3 is 322.18, found 322.60.

[0192] 2-(Methyl(4-morpholino-4a,7a-dihydro-7 / - / -pyrrolo[2,3-c / ]pyrimidin-2-yl)amino)ethan-1-ol (DPM-A24):S-T00387W0001 32917 / 70090 / PC

[0193] A mixture of 7 (100.0 mg, 0.41 mmol), 2-(methylamino)ethan-1-ol (0.03 mL, 0.41 mmol), and N, N-diisopropylethylamine (145.0 L, 0.83 mmol) in N-methylpyrrolidone (1.0 mL) in the sealed vial was stirred at 160 °C for 3h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A24 (53.0 mg, 46% yield). MS(m / z): [M+H] calc'd for C13H21N5O2 is 278.15, found 278.30.

[0194] (R)-1-(4-morpholino-7H-pyrrolo[2,3-d]pyrimidin-2-yl)pyrrolidin-3-ol (DPM-A42):OH

[0195] A mixture of 7 (500.0 mg, 0.21 mmol), (R)-py rrolidi n-3-ol (91.0 mg, 1.0 mmol), and N, N-diisopropylethylamine (73.0 pL, 0.83 mmol) in 1- butanol (1.0 mL) in the sealed vial was stirred in microwave at 160 °C for 3h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A42 (23.0 mg, 38% yield). MS(m / z): [M+H] calc’d for C14H19N5O2 is 290.15, found 290.30.

[0196] 2-(ethyl(4-morpholino-7 / - / -pyrrolo[2,3-c / |pyrimidin-2-yl)amino)ethan-1-ol (DPM-A43)

[0197] A mixture of 7 (50.0 mg, 0.21 mmol), 2-(ethylamino)ethan-1 -ol (37.0 mg, 0.42 mmol), and N, N-diisopropylethylamine (73.0 pL, 0.42 mmol) in 1 -butanol (1.0 mL) in the sealed vial was stirred in a microwave at 160 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A43 (21.0 mg, 34% yield). MS(m / z): [M+H] calc’d for C14H21N5O2 is 292.17, found 292.70.S-T00387W0001 32917 / 70090 / PC

[0198] 3-(methyl(4-morpholino-7 / - / -pyrrolo[2,3-c / ]pyrimidin-2-yl)amino)propan-1-ol (DPM-A44)0

[0199] A mixture of 7 (50.0 mg, 0.21 mmol), 3-(methylamino)propan-1-ol (37.0 mg, 0.42 mmol), and N,N-diisopropylethylamine (73.0 pL, 0.42 mmol) in 1 -butanol (1.0 mL) in the sealed vial was stirred in a microwave at 160 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A44 (15.0 mg, 25% yield). MS(m / z):[M+H] calc’d for C14H21N5O2 is 292.17, found 292.70.

[0200] 2-methyl-1-(methyl(4-morpholino-7 / - / -pyrrolo[2,3-c / ]pyrimidin-2-yl)amino)propan-2-ol (DPM-A45)

[0201] A mixture of 7 (50.0 mg, 0.21 mmol), 2-methyl-1-(methylamino)propan-2-ol (43.0 mg, 0.42 mmol), and N, N-diisopropylethylamine (73.0 pL, 0.42 mmol) in 1 -butanol (1.0 mL) in the sealed vial was stirred in a microwave at 160 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A45 (20.0 mg, 31% yield). MS(m / z): [M+H] calc’d for C15H23N5O2 is 306.19, found 306.40.

[0202] (R)-3-methyl-1-(4-morpholino-7H-pyrrolo[2,3-c / ]pyrimidin-2-yl)pyrrolidin-3-ol (DPM-A48)OH

[0203] A mixture of 7 (25.0 mg, 0.10 mmol), (R)-3-methylpyrrolidin-3-ol (21.0 mg, 0.21 mmol), and N,N-diisopropylethylamine (36.0 pL, 0.21 mmol) in 1 -butanol (1.0 mL) in the sealed vial was stirred in a microwave at 160 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and thenS-T00387W0001 32917 / 70090 / PCpurified by silica gel chromatography, resulting in the pure product DPM-A48 (15.0 mg, 47% yield). MS(m / z):[M+H] calc’d for C15H21N5O2 is 304.17, found 304.70.

[0204] (3R,4R)-1-(4-morpholino-7H-pyrrolo[2,3-c]pyrimidin-2-yl)pyrrolidine-3,4-diol (DPM-A49)OH

[0205] A mixture of 7 (25.0 mg, 0.10 mmol), (3R,4R)-pyrrolidine-3,4-diol (22.0 mg, 0.21 mmol), and N, N-diisopropylethylamine (36.0 pL, 0.21 mmol) in 1 -butanol (1.0 mL) in the sealed vial was stirred in a microwave at 160 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A49 (13.0 mg, 41% yield). MS(m / z):[M+H] calc’d for C14H19N5O3 is 306.15, found 306.30.

[0206] ((3S,4S)-1-(4-morpholino-7 / - / -pyrrolo[2,3-c / ]pyrimidin-2-yl)pyrrolidine-3,4-diol (DPM-A50)OH

[0207] A mixture of 7 (25.0 mg, 0.10 mmol), (3S,4S)-pyrrolidine-3,4-diol (22.0 mg, 0.21 mmol), and N, N-diisopropylethylamine (36.0 pL, 0.21 mmol) in 1 -butanol (1.0 mL) in the sealed vial was stirred in a microwave at 160 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A50 (17.0 mg, 53% yield). MS(m / z):[M+H] calc’d for C14H19N5O3 is 306.15, found 306.50.

[0208] 4,4-difluoro-1-(4-morpholino-7H-pyrrolo[2,3-c / ]pyrimidin-2-yl)pyrrolidin-3-ol (DPM-A51)S-T00387W0001 32917 / 70090 / PCON

[0209] A mixture of 7 (25.0 mg, 0.10 mmol), 4,4-difluoropyrrolidin-3-ol (26.0 mg, 0.21 mmol), and N, N-diisopropylethylamine (36.0 pL, 0.21 mmol) in 1 -butanol (1.0 mL) in the sealed vial was stirred in a microwave at 160 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A51 (14.0 mg, 41% yield). MS(m / z):[M+H] calc’d for C14H17F2N5O2 is 326.14, found 326.70.

[0210] (3S,4S)-4-fluoro-1-(4-morpholino-7H-pyrrolo[2,3-c / ]pyrimidin-2-yl)pyrrolidin-3-ol (DPM-A52)

[0211] A mixture of 7 (25.0 mg, 0.10 mmol), (3S,4S)-4-fluoropyrrolidin-3-ol (22.0 mg, 0.21 mmol), and N, N-diisopropylethylamine (36.0 pL, 0.21 mmol) in 1 -butanol (1.0 mL) in the sealed vial was stirred in a microwave at 160 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A52 (16.0 mg, 50% yield). MS(m / z):[M+H] calc’d for C14H18FN5O2 is 308.14, found 308.70.

[0212] 3-ethyl-1-(4-morpholino-7 / - / -pyrrolo[2,3-c / ]pyrimidin-2-yl)azetidin-3-ol (DPM-A53)

[0213] A mixture of 7 (25.0 mg, 0.10 mmol), 3-ethylazetidin-3-ol (21.0 mg, 0.21 mmol), and N, N-diisopropylethylamine (36.0 pL, 0.21 mmol) in 1 -butanol (1.0 mL) in the sealed vial was stirred in a microwave at 160 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and thenS-T00387W0001 32917 / 70090 / PCpurified by silica gel chromatography, resulting in the pure product DPM-A53 (13.0 mg, 41% yield). MS(m / z):[M+H] calc’d for C15H21N5O2 is 304.17, found 304.30.

[0214] (S)-3-methyl-1-(4-morpholino-7 / - / -pyrrolo[2,3-c / ]pyrimidin-2-yl)pyrrolidin-3-ol (DPM-A54)

[0216] A mixture of 7 (25.0 mg, 0.10 mmol), (S)-3-methylpyrrolidin-3-ol (21.0 mg, 0.21 mmol), and N, N-diisopropylethylamine (36.0 pL, 0.21 mmol) in 1 -butanol (1.0 mL) in the sealed vial was stirred in a microwave at 160 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A54 (14.0 mg, 44% yield). MS(m / z):[M+H] calc’d for C15H21N5O2 is 304.17, found 304.60.

[0217] 1-(4-morpholino-7 / - / -pyrrolo[2,3-c / ]pyrimidin-2-yl)-3-phenylpyrrolidin-3-ol (DPM-A55)

[0218] A mixture of 7 (25.0 mg, 0.10 mmol), racemic 3-phenylpyrrolidin-3-ol (34.0 mg, 0.21 mmol), and N, N-diisopropylethylamine (36.0 pL, 0.21 mmol) in 1 -butanol (1.0 mL) in the sealed vial was stirred in a microwave at 160 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A55 (13.0 mg, 34% yield). MS(m / z):[M+H] calc’d for C20H23N5O2 is 366.19, found 366.80.

[0219] 1-(4-morpholino-7 / - / -pyrrolo[2,3-c / ]pyrimidin-2-yl)azetidin-3-ol (DPM-A56)S-T00387W0001 32917 / 70090 / PC

[0220] A mixture of 7 (100.0 mg, 0.42 mmol), azetidin-3-ol (153.0 mg, 2.1 mmol), and N, N-diisopropylethylamine (146.0 pL, 0.84 mmol) in 1-butanol (1.0 mL) in the sealed vial was stirred in a microwave at 160 °C for 3 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A56 (15.0 mg, 13% yield). MS(m / z): [M+H] calc’d for C13H17N5O2 is 276.14, found 276.30.

[0221] Scheme 5. Synthetic route to the synthetic intermediate 4-(2-chloro-7-(2-methoxyethyl)-7 / - / -pyrrolo[2,3-d]pyrimidin-4-yl)morpholine (9).ClPPh3, DIAD K2CO3THF, 25°C, 22h THF, 0°C, 1h

[0222] To a stirred solution of PPhs (828.0 mg, 3.16 mmol) and 2,4-dichloro-4a,7a-dihydro-7 / - / -pyrrolo[2,3-d]pyrimidine (300.0 mg, 1.58 mmol) in THF (5.0 mL), 2-methoxyethan-1-ol (0.1 mL, 1.26 mmol) was added at room temperature. The reaction mixture was then cooled to 0°C and diisopropyl azodicarboxylate (0.6 mL, 3.16 mmol) was added dropwise. The resulting mixture was stirred at room temperature for 22 hours, after which the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, resulting in the pure product compound 8. To a mixture of 8 (75.0 mg, 0.30 mmol) in 3 mL of THF, potassium carbonate (84.0 mg, 0.60 mmol) and morpholine (0.03 mL, 0.30 mmol) were added, and the reaction mixture was stirred at 0°C for 3 hours. Compound 9 was obtained in quantitative yield and purified by silica gel chromatography. MS(m / z): [M+H] calc'd for C13H17CIN4O2 is 297.10, found 297.90.

[0223] 2,2'-((7-(2-Methoxyethyl)-4-morpholino-7H-pyrrolo[2,3-d|pyrimidin-2-yl)azanediyl)bis(ethan-1-ol) (DPM- A28):

[0224] A mixture of 9 (110.0 mg, 0.37 mmol), diethanolamine (0.08 mL, 0.74 mmol), and N, N-diisopropylethylamine (129.0 pL, 0.74 mmol) in 1-butanol (2.0 mL) was stirred in microwave at 130 °C for 1.5 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gelS-T00387W0001 32917 / 70090 / PCchromatography, resulting in the pure product DPM-A28 (51.0 mg, 38% yield). MS(m / z): [M+H] calc'd for C17H27N5O4 is 366.21, found 366.30.

[0225] 2-((7-(2-Methoxyethyl)-4-morpholino-7H-pyrrolo[2,3-d]pyrimidin-2-yl)(methyl)amino)ethan-1-ol (DPM- A29):

[0226] A mixture of 9 (55.0 mg, 0.19 mmol), 2-(methylamino)ethan-1-ol (0.03 mL, 0.37 mmol), and N, N-diisopropylethylamine (32.0 pL, 0.37 mmol) in 1 -butanol (2.0 mL) was stirred in microwave at 130 °C for 1.5 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A29 (33.0 mg, 53% yield). MS(m / z): [M+H] calc'd for C16H25N5O3 is 336.20, found 336.80.

[0227] Scheme 6. Synthetic route to the synthetic intermediate 4-(2-chloro-4,5-dihydro-9H-purin-6-yl)morpholine (11):ClPhSO3H, EtOAc 50 °C, 1 h

[0228] 4-(2-chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine (11). A mixture of 2,6-dichloropurine (1.0 g, 5.2 mmol) and benzenesulfonic acid (11.0 mg, 0.062 mmol) in ethyl acetate (20 mL) was heated to 50 °C under argon. 3,4-dihydro-2H-pyran (0.8 mL, 8.8 mmol) was then added dropwise in ethyl acetate at a rate of 1 mL per minute. The mixture was stirred for 1 hour after the addition. Compound 10 was obtained by filtration and subsequently treated with morpholine under microwave irradiation at 100 °C, resulting in compound 11. MS(m / z):[M+H] calc’d for C14H18CIN5O2 is 324.11, found 324.30.

[0229] 2,2'-((6-morpholino-9H-purin-2-yl)azanediyl)bis(ethan-1-ol) (DPM-A26):S-T00387W0001 32917 / 70090 / PCOH

[0230] A mixture of 11 (200.0 mg, 0.6 mmol), diethanolamine (0.1 mL, 1.24 mmol), and N, N-diisopropylethylamine (215.0 pL, 1.24 mmol) in NMP (2.0 mL) was stirred in microwave at 160 °C for 1.5 hours. After completion, the reaction mixture was concentrated to dryness under vacuum and the residue was reacted with HCI (4 M, 1 mL) for overnight. The reaction mixture was purified by silica gel chromatography, resulting in the pure product DPM-A26 as its HCI salt (84.0 mg, 53% yield). MS(m / z): [M+H] calc'd for C13H20N6O3 is 309.16, found 309.30.

[0231] 2-(methyl(6-morpholino-9H-purin-2-yl)amino)ethan-1-ol (DPM-A27):

[0232] A mixture of 11 (200.0 mg, 0.6 mmol), 2-(methylamino)ethan-1-ol (0.1 mL, 1.24 mmol), and N, N-diisopropylethylamine (0.2 mL, 1.24 mmol) in NMP (2.0 mL) was stirred in microwave at 160 °C for 1.5 hours. After completion, the reaction mixture was concentrated to dryness under vacuum and the residue was reacted with HCI (4 M, 1 mL) an overnight. The reaction mixture was purified by silica gel chromatography, resulting in the pure product DPM-A27 as its HCI salt (78.0 mg, 51% yield). MS(m / z): [M+H] calc'd for C12H18N6O2 is 279.15, found 279.80.

[0233] Scheme 7. Synthetic route to 4-(2-chloro-9-(2-methoxyethyl)-9H-purin-6-yl)morpholine (13).ClPPh3, DIAD K2CO3THF, RT, 22h THF, RT, 1h

[0234] Compounds 12 and 13 were obtained using the general procedure used to make compounds 8 and 9.Compound 13, MS(m / z): [M+H] calc'd for C12H16CIN5O2 is 298.10, found 298.30.S-T00387W0001 32917 / 70090 / PC

[0235] 2,2'-((9-(2-methoxyethyl)-6-morpholino-9H-purin-2-yl)azanediyl)bis(ethan-1-ol) (DPM-A30):

[0236] A mixture of 13 (38.0 mg, 0.13 mmol), diethanolamine (0.03 mL, 0.26 mmol), and N, N-diisopropylethylamine (44.0 pL, 0.26 mmol) in 1 -butanol (2.0 mL) was stirred in microwave at 130 °C for 1.5 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A30 (15.0 mg, 32% yield). MS(m / z): [M+H] calc'd for C16H26N6O4 is 367.20, found 367.30.

[0237] 2,2'-((9-(2-hydroxyethyl)-6-morpholino-9H-purin-2-yl)azanediyl)bis(ethan-1-ol) (DPM-A31):

[0238] A mixture of DPM-A26 (50.0 mg, 0.16 mmol), 2-bromoethan-1-ol (26.0 mg, 0.21 mmol), and sodium hydride (8.4 mg, 0.21 mmol) in DMF (2.0 mL) was stirred at room temperature for an overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A31 (22.0 mg, 38% yield). MS(m / z): [M+H] calc'd for C15H24N6O4 is 353.19, found 353.40.

[0239] 2-(2-((2-hydroxyethyl)(methyl)amino)-6-morpholino-9H-purin-9-yl)ethan-1-ol (DPM-A32):HOS-T00387W0001 32917 / 70090 / PC

[0240] A mixture of DPM-A27 (50.0 mg, 0.18 mmol), 2-bromoethan-1-ol (29.0 mg, 0.23 mmol), and sodium hydride ( 9.3 mg, 0.23 mmol) in DMF (2.0 mL) was stirred at room temperature for an overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A32 (20.0 mg, 35% yield). MS(m / z): [M+H] calc'd for C14H22N6O3 is 323.18, found 323.60.

[0241] Scheme 8. Synthetic route to 2,2'-((4-morpholinoquinazolin-2-yl)azanediyl)bis(ethan-1 -ol) (DPM-A25):EtN(iPr)2NMP, 160°C, 1.5h

[0242] A mixture of 2,4-dichloroquinazoline (1.0 g, 5.0 mmol) and potassium carbonate (1.0 g, 10.0 mmol) in THF was kept at -78°C. Morpholine (0.4 g, 5 mmol) was then added dropwise in THF at a rate of 1 mL per minute. The mixture was stirred for 1 hour after the addition. The reaction mixture was concentrated to dryness and compound 14 was isolated by silica gel chromatography. The mixture of 14 (50.0 mg, 0.20 mmol), diethanolamine (0.06 mL, 0.6 mmol) and N, N-diisopropylethylamine (35.0 pL, 0.20 mmol) in NMP (2.0 mL) was stirred in microwave at 160 °C for 1.5 h. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A25 (15.0 mg, 24% yield). (MS(m / z): [M+H] calc’d for C16H22N4O3 is 319.17, found 319.30.

[0243] Scheme 9. Synthetic route to the synthetic intermediates 2-chloro-4-(piperidin-1 -yl)pyrimidine (15) and 4-(2-chloropyrimidin-4-yl)morpholine (16):ClK2CO3THF, -78°C, 1h15 (X=CH2)16 (X=O)

[0244] Compounds 15 and 16 were synthesized and purified using the general procedure for compound 1.

[0245] 2,2'-((4-(piperidin-1-yl)pyrimidin-2-yl)azanediyl)bis(ethan-1-ol) (DPM-A33):S-T00387W0001 32917 / 70090 / PCOH

[0246] A mixture of 15 (40.0 mg, 0.2 mmol), diethanolamine (0.06 mL, 0.6 mmol), and potassium carbonate (42.0 mg, 0.3 mmol) in THF (2.0 mL) was stirred at room temperature overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and purified using silica gel chromatography, yielding the pure product DPM-A33 (25.0 mg, 46% yield). MS(m / z): [M+H] calc'd for C13H22N4O2 is 267.17, found 267.70.

[0247] 2,2'-((4-morpholinopyrimidin-2-yl)azanediyl)bis(ethan-1-ol) (DPM-A37):OH

[0248] A mixture of 16 (30.0 mg, 0.15 mmol), diethanolamine (0.08 mL, 0.75 mmol), and potassium carbonate (31.0 mg, 0.23 mmol) in THF (2.0 mL) was stirred at room temperature overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and purified using silica gel chromatography, yielding the pure product DPM-A37 (15.0 mg, 37% yield). MS(m / z): [M+H] calc'd for C12H20N4O3 is 269.15, found 269.80.

[0249] 2-(methyl(4-morpholinopyrimidin-2-yl)amino)ethan-1-ol (DPM-A38):

[0250] A mixture of 16 (30.0 mg, 0.15 mmol), 2-(methylamino)ethan-1-ol (0.06 mL, 0.75 mmol), and potassium carbonate (31.0 mg, 0.23 mmol) in THF (2.0 mL) was stirred at room temperature overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and purified using silica gel chromatography, yielding the pure product DPM-A38 (20.0 mg, 56% yield). MS(m / z): [M+H] calc'd for Ci 1 H18N4O2 is 239.14, found 239.20.S-T00387W0001 32917 / 70090 / PC

[0251] Scheme 10. Synthetic route to the synthetic intermediate 4-(2-chloro-7-(2-methoxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)morpholine (18).ciK2CO3K2CO3ci THF, -78° C, 1h THF, 0°C, 1h

[0252] Perchloropy rimido[5,4-d]py ri midine (1.0 g, 4.0 mmol) and potassium carbonate (1.0 g, 7.0 mmol) in THF was kept at -78°C. Diethylamine (0.3 g, 4.0 mmol) was then added dropwise in THF at a rate of 1 mL per minute. The mixture was stirred for 1 hour after the addition. Compound 17 was obtained as a yellow powder, in quantitative yield. To a mixture of 17 (500.0 mg, 1.33 mmol) in lO mL of THF, diethanolamine (0.15 mL) was added, and the reaction mixture was stirred at 0°C for 1 hour. Compound 18 was obtained in quantitative yield and purified using a similar work-up procedure as compound 2. MS(m / z): [M+H] calc'd for C14H20CI2N6O2 is 375.10, found 375.73.

[0253] Scheme 11. Synthetic route to 2,2'-((8-(Diethylamino)-2-morpholino-6-(piperidin-1-yl)pyrimido[5,4-d]pyrimidin-4-yl)azanediyl)bis(ethan-1-ol) (DPM-A1).EtN(iPr)2p-dioxane 100°C, 12h

[0254] Compound 19 was synthesized by treating compound 18 (50.0 mg, 0.13 mmol) with piperidine (0.013 mL) and potassium carbonate (55.0 mg, 0.40 mmol) in THF at room temperature for 1 hour. The reaction mixture was purified following similar work-up as compound 2 to obtain compound 19 (41.0 mg, 73% yield). MS(m / z):[M+H] calc'd for C19H30CIN7O2 is 424.21, found 423.89. A mixture of 19 (35.0 mg, 0.083 mmol), morpholine (0.02 mL, 0.25 mmol), and N, N-diisopropylethylamine (29.0 pL, 0.17 mmol) in p-dioxane (2.0 mL) was stirred at 100 °C overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A1 (19.0 mg, 48% yield). MS(m / z):[M+H] calc’d for C23H38N8O3is 475.31, found 475.17.

[0255] Scheme 12. Synthetic route to 2,2'-((8-(diethylamino)-6-morpholino-2-(piperidin-1 -yl)pyrimido[5,4-d]pyrimidin-4-yl)azanediyl)bis(ethan-1-ol) (DPM-A4):S-T00387W0001 32917 / 70090 / PCEtN(iPr)2p-dioxane, 100°C, 12h

[0256] Compound 20 was synthesized by treating compound 18 (50 mg, 0.13 mmol) with morpholine (0.01 mL) and potassium carbonate (55 mg, 0.40 mmol) at room temperature for 1 hour. The reaction mixture was purified following a similar work-up procedure as compound 2 to obtain compound 20 (35.0 mg, 62% yield).MS(m / z): [M+H] calc'd for C18H29CIN7O3 is 426.19, found 426.11. A mixture of 20 (30.0 mg, 0.07 mmol), piperidine (0.02 mL, 0.21 mmol), and N, N-diisopropylethylamine (25.0 pL, 0.14 mmol) in p-dioxane (2.0 mL) was stirred at 100 °C for overnight. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A4 (17.0 mg, 51% yield).MS(m / z): [M+H] calc’d for C23H38N8O3 is 475.31, found 475.18.

[0257] Scheme 13. Synthetic route to the synthetic intermediate 4,4'-(2,6-dichloropyrimido[5,4-d]pyrimidine-4,8-diyl)bis(2-methylmorpholine) (21):K2CO3THF, 25°C, 3h

[0258] A mixture of perchloropyrimido[5,4-d]pyrimidine (300.0 mg, 1.1 mmol) and potassium carbonate (307.0 mg, 2.2 mmol), and (+)2-methylmorpholine (337.0 mg, 3.3 mmol) in THF was stirred at room temperature for 3 hours. Water was added to form a yellow precipitate. The precipitate was dissolved in ethyl acetate, filtered, concentrated under vacuum, and a yellow glassy solid was obtained. The solid was triturated with diethyl ether, filtered, and dried to yield a yellow powder, compound 21, in quantitative yield. MS(m / z): [M+H] calc'd for C16H20CI2N6O2 is 399.10, found 399.40.

[0259] 2,2',2",2"'-((4,8-bis(2-methylmorpholino)pyrimido[5,4-d]pyrimidine-2,6-diyl)bis(azanetriyl))tetrakis(ethan- 1-ol) (DPM-A39):S-T00387W0001 32917 / 70090 / PC

[0260] A mixture of 21 (50.0 mg, 0.13 mmol), diethanolamine (0.1 mL, 1.3 mmol), and N,N-diisopropylethylamine (44.0 pL, 0.25 mmol) in 1 -butanol (1.0 mL) was stirred in the microwave at 160 °C for 7 hours. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by silica gel chromatography, resulting in the pure product DPM-A39 (28.0 mg, 42% yield). MS(m / z): [M+H] calc'd for C24H40N8O6 is 537.31, found 537.70.

[0261] (3R,3'R)-1, T-(4,8-bis(2-methylmorpholino)pyrimido[5,4-d]pyrimidine-2,6-diyl)bis(pyrrolidin-3-ol) (DPM-

[0262] A mixture of 21 (50.0 mg, 0.13 mmol), (R)-pyrrolidin-3-ol (55.0 mg, 0.6 mmol), and N,N-diisopropylethylamine (44.0 pL, 0.25 mmol) in 1 -butanol (1.0 mL) was stirred in the microwave at 160 °C for 3 hours. After completion, the reaction mixture was concentrated to dryness under vacuum and then purified by C18 silica gel chromatography, resulting in the pure product DPM-A40 (25.0 mg, 40% yield). MS(m / z): [M+H] calc'd for C24H36N8O4 is 501.29, found 501.80.

[0263] 2-(methyl(6-methyl-4-morpholino-7H-pyrrolo[2,3-d]pyrimidin-2-yl)amino)ethan-1-ol (DPM-A57).This compound is prepared following the same general procedure for the preparation of DPM-A24, but using 2,4-dichloro-6-methy l-7H-pyrrolo[2,3-d]pyrimidine as the starting material. MS(m / z): [M+H] calc'd for C14H21N5O2 is 292.17, found 292.70.ciS-T00387W0001 32917 / 70090 / PC

[0264] 2-(methyl(5-methyl-4-morpholino-7H-pyrrolo[2,3-d]pyrimidin-2-yl)amino)ethan-1-ol (DPM-A58).This compound is prepared following the same general procedure for the preparation of DPM-A24, but using 2,4-dichloro-5-methy l-7H-pyrrolo[2,3-d]pyrimidine as the starting material. MS(m / z): [M+H] calc'd for C14H21N5O2 is 292.17, found 292.70..o.H H

[0265] 2-(methyl(4-thiomorpholino-7H-pyrrolo[2,3-d]pyrimidin-2-yl)amino)ethan-1-ol (DPM-A59). This compound is prepared following the same general procedure for the preparation of DPM-A24, but using thiomorpholine in place of morpholine. MS(m / z): [M+H] calc'd for C13H19N5OS is 294.13, found 294.70. The corresponding sulfone (DPM-A60) and sulfoxide (DPM-A61) are obtained by oxidation of this compound using conventional means.NHBIOLOGICAL TESTING METHODSDifferentiation of lower motor neurons from human isogenic and ALS iPSCs

[0266] The IPSO clones (ALS-C9orf72 patient IPSO and their isogenic control; ALS-TDP43 (M337V) engineered IPSO and its isogenic control; and ALS-SOD1 (A5V) engineered IPSO and its isogenic control) employed contain a stably integrated vector (hNIL; CLYBL-TO-hNIL-BSD-mApple), a gift from Michael Ward (Addgene plasmid # 124230; http: / / n2t.net / addgene: 124230), that encodes the transcription factors (NGN2, ISL1, LHX3) necessary for differentiation of IPSCs into lower motor neurons (MN) (Mazzoniet al., 2013). The transcription factor genes were located behind the tetracycline response element (TRE3G) that is inducible with doxycycline. The vector also contained a GAG promoter driving the constitutive expression of the reverse tetracycline transactivator (rtTA3C). The vector was inserted at the CLYBL safe harbor locus (Cerbini et al., 2015) in the IPSO genome.

[0267] The IPSCs were differentiated into lower motor neurons (MN) as described in Fernandopulle et al. (Fernandopulle et al., 2018), with modifications. In brief, the IPSCs were dissociated with accutase into single cells and plated in Matrigel-coated plates (for 6-well plate: seed 5-5.5x10A5 cells / well; for 10-cm dishes: seed 3x10A6; for 15-cm dishes: 5- 7x10A6) in mTeSR plus medium containing 10 pM ROCK inhibitor (plating day is day -1). Twenty-four hours later (day 0), cells were induced with Neural Induction Medium (NIM): DMEM / F12 with HEPES and glutamine (gin), B27 supplement (1x), MEM Non-Essential Amino Acids (1x, NEAA), GlutaMAXS-T00387W0001 32917 / 70090 / PC(1x), compound E (200 nM), ROCK inhibitor (10 pM), doxycycline (2 pg / mL) and Culture One (1x). On day 1 of MN differentiation, media was removed and replaced with fresh NIM media containing 2 pig / mL doxycycline. On day 2 of MN differentiation, the cells began showing signs of MN differentiation by beginning to extend axons and dendrites (neurites). These MN precursors (MNP) were dissociated with accutase to single cells, strained through 40 pirn cell strainer and frozen in Bambanker freezing media (3x106D2 MNPs / cryovial aliquot).

[0268] Day 2 MNPs were plated on poly-D-lysine (PDL) / laminin coated 384-well plates along with lentivirus expressing mitochondrial (MT)-targeted GFP to visualize MT, and a cytoplasmically-targeted mScarlet to visualize somas and neurites. For this, Day 2 MNPs were thawed and replated (10,500 D2 MNPs / well in 384-well plates, volume 40 pil) with CAG-mtTagGFP2-2A-mScarlet lentivirus at a multiplicity of infection (MOI)=4 in MN differentiation medium (MNDM): half DMEM / F12 with HEPES and Gin and half Neurobasal Plus neuronal medium, B27 Plus (1x), NEAA (1 x), GlutaMAX (1x), Penicillin / Streptomycin 5000 U (1x), compound E (200 nM), doxycycline (dox, 2 pg / mL), laminin (1 pg / mL), BDNF (10 ng / mL), GDNF (10 ng / mL), NT3 (10 ng / mL) and Culture One (1 x). On day 4, 20 pil of the media was removed and an additional 60 pil of fresh MN culture medium was added bringing the total final volume to 80 pil. The MN culture medium contained Neurobasal Plus neuronal medium, B27 Plus supplement (1x), NEAA (1x), GlutaMAX (1x), Penicillin / Streptomycin 5000 U (1x), laminin (1 pg / mL), BDNF (10 ng / mL), GDNF (10 ng / mL), and NT3 (10 ng / mL). Every 4 days half of the cell culture medium was removed and replaced by fresh MN culture media. Doxycycline (2 pg / mL) was added only for the day 4 media change and Culture one (1 x) for day 4 and day 8 media changes.Differentiation of glutamatergic cortical neurons from human isogenic and AD iPSCs

[0269] The AD PS1-A246E and its ISO IPSC were differentiated into glutamatergic cortical neurons (CN) using the pLVX-UbC-rtTA-Ngn2-2A-Ascl (UNA) plasmid as described (MacMullen et al., 2025) and subsequently plated into 384w microtiter plates. MT and neurites were labeled by infection with a lentivirus that expresses a pCAG-mtTAGGFP2-2A-mScarlet plasmid and the neurons subsequently assayed using the longitudinal mitochondrial dynamics assay. Alternatively, they were plated in 96w Seahorse Bioanalyzer plates without virus and assayed for MT bioenergetics.Longitudinal mitochondrial (MT) dynamics assay

[0270] After plating ALS diseased or isogenic control motor neurons (MN), or AD diseased or isogenic control cortical neurons (CN), in 384w microtiter plates, MN were imaged every two days, beginning at day 8, until day 30 to follow the MT and neurite metrics across time. Images (four fields per well in 384w plate) were captured using the Image Xpress Micro Confocal High-Content Imaging System (at 37°C, 5% CO2) with 60X objective, 0.95 NA with 60pim pinhole. Each field was acquired as a z-stack of five individual images separated by 0.7 pim and maximum intensity projections of each individual stack was used for analysis. A custom segmentation algorithm was applied to each image to segment MT and neurites using the MetaXpress software v.6.7.2.290. The following parameters were collected from these images: total MT count, median MT length and neurite cumulative area (CA). CA is the number of pixels representing neurites in each field and provides a metric forS-T00387W0001 32917 / 70090 / PCneurite complexity and neurite degeneration. Custom python codes were used to extract the MT and neurite data from these field images and four fields per well were averaged to yield an average per well for each parameter. Experimental average data for each parameter were calculated based on genotype and treatment (with or without compounds). The averaged data for each MT and neurite parameter from each of the time course experiments were then averaged together for each time point tested. A two-way repeated measure ANOVA, followed by Bonferroni's or Tukey's multiple comparisons test was performed to calculate the statistical significance between the groups at each time point using GraphPad Prism (version 10.2.3).Compound addition, time course and dose response (D: R) assays

[0271] The Day 2 MNPs [ALS (09, TDP43, or SOD1) and the isogenic (ISO) control] were thawed and replated with CAG-mtTagGFP2-2A-mScarlet lentivirus at a MOM in PDL / laminin coated 384-well plates at a cell density of 10,500 MNPs in 40 pL of MNDM media containing 2 pig / mL dox / well. On day 4, 20 piL of media with lentivirus was removed per well, and 60 piL of fresh culture media with 2 pig / mL dox was added (total volume 80 piL). AD (PS1) ON and the isogenic control were plated in a similar fashion (MacMullen et al., 2025). The plates were imaged as described above on days 8, 10 and 12. To test the selected compounds, ALS MNs or AD ON and corresponding ISO neurons on day 10 / day 12, were treated with DMSO alone (0.125% (v / v), 20 - 40 technical replicates) or compounds dissolved in DMSO at different concentrations (7- or 10-point D: R assays with 5 technical replicates). Prior to adding compounds, 40 piL of media was removed per well, followed by addition of 20 piL of fresh culture media and then 20 piL of compound at the appropriate concentrations. Compounds (in DMSO) were placed in a source 384-well plate and then diluted in culture media twice, ensuring that the DMSO concentration remained constant at 0.125% in all wells. Compounds were only added once at day 10 / day 12 after imaging, and plates were imaged every two days until day 30, as described above. Generally, three to five independent time course experiments were carried out for the selected compounds. D: R experiments (across days) were used for measuring the potency (EC50) and efficacy (span) of the compounds. For the D: R experiments, robust z-scores were calculated relative to the in-plate DMSO control and were averaged for each compound across replicate plates.Motor neuron viability assay

[0272] MN viability in response to DPM (20 piM - 156 nM), was evaluated using the RealTime-Glo MT Cell Viability Assay (Promega) according to the manufacturer's protocol. Viable cells with active metabolism reduce the pro-substrate (MT Cell Viability substrate) into a substrate, which diffuses into the culture medium and is used by NanoLuc luciferase to generate a luminescent signal. The signal corresponds to the number of viable cells. Dead cells do not reduce the pro-substrate and therefore produce no signal. Briefly, NanoLuc luciferase enzyme and a cell-permeant pro-substrate, were added directly to the media of the MNs in culture in 384-well plates on day 31 and luminescence measured on day 33 and day 36 by CLARIOstar microplate reader (BMG Labtech). A one-way ANOVA, followed by Tukey's multiple comparisons test was performed to calculate the statistical significance between the groups using GraphPad Prism (version 10.2.3).S-T00387W0001 32917 / 70090 / PCMT bioenergetics (Seahorse analyzer assay)

[0273] Seahorse plates were coated with 0.1 mg / mL PDL overnight at 37 °C. Plates were washed with sterile water, air-dried and further coated with 15 g / mL laminin at 37 °C for 2 hours. Day 2 MNPs or CNPs were plated on the PDL / laminin coated Seahorse plates at a density of 30,000 cells / well in a total volume of 180 piL of differentiation medium (MNDM). A total of 15 replicate wells per neuron type were plated for each experiment. Two days after plating, 135 piL media was removed and replaced with an equal volume of fresh media. 50% media change was performed every 4 days until the day of the assay. Compound was added on day 8 of culture and the MT functional tests were performed on day 14 using a Seahorse XFe96 analyzer.

[0274] On day 14, 1h prior to the assay, the media was replaced with Seahorse XF DMEM media supplemented with 10 mM glucose, 2 mM glutamine, and 1 mM pyruvate and the plate was incubated at 37°C in a CO2free incubator. The injection ports of a hydrated Seahorse cartridge were filled with 2 piM oligomycin, 4 piM carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone (FCCP), and 1 pi M each of rotenone and antimycin A, and the cartridge was calibrated. Following calibration, the cell culture plate was placed in the Seahorse analyzer and the oxygen consumption rates (OCR) were measured. For each injection, 3 OCR measurements were recorded. The OCR values were normalized to the number of live cells in each well. Live cell numbers were determined by calcein AM staining. After the final OCR measurement, calcein AM was added to each well at a final concentration of 10 piM. The assay plate was incubated in a CO2free incubator at 37°C for 30 mins and live cell counts were obtained using an InCell 6000 analyzer (GE Healthcare Technologies).

[0275] Four test parameters were analyzed: basal respiration, the general resting state respiratory capacity of the neurons; ATP-linked OCR, an estimation of the amount of energy able to be produced by neurons which is measured after the oligomycin injection; maximal respiration, the amount of energy released when challenged with the electron transport chain uncoupler FCCP; and spare respiratory capacity, the difference between maximal and basal respiration values which indicates the neuron's energy reserve under periods of stress. Average values of the 15 replicate wells were derived for each experiment for all the four MT respiratory parameters.Pyruvate Uptake Assay

[0276] Pyruvate uptake into mitochondria was measured using the following protocol. Fresh mitochondria were isolated from the liver of 4-month-old male C57BL6 / J mice. They were suspended in uptake buffer with either UK-5099 (a known MPC inhibitor, used as a control) or test compound (e.g., DPM, DPM-A23), both in dose response. Subsequently, the substrate mix containing C14-pyruvate was added and after the appropriate time the reaction was stopped. The mitochondria were purified by centrifugation, resuspended in lysis buffer, and the amount of C14-pyruvate taken up during the reaction was determined by scintillation counting. Raw C14counts per minute of assay were converted to pmol pyruvate / mg protein using a protein quantification assay. The results are shown in Figure 16.S-T00387W0001 32917 / 70090 / PCRESULTS FROM BIOLOGICAL EXPERIMENTSEfficacy in C9orf72-ALS Motor Neurons

[0277] A collection of approximately 14,400 compounds (Varkuti et al., 2020a, b; MacMullen and Davis, 2021; unpublished) was assayed for the ability to preserve MT health, as indicated by visually apparent parameters, such as MT count, MT length, neurite cumulative area, and persistence of motor neuron viability. Each of these first 4 parameters is severely compromised in affected motor neurons in ALS patients before the neurons die and can also be observed in neuronal culture, using ALS C9orf72 iPSC-derived lower motor neurons (C9 MN). A protocol was used wherein the neurons in individual wells were cultured for 12 days and then each well was exposed to a test compound or DMSO control on day 12. The C9 MN begin to show signs of mitochondrial dysfunction by day 14 and typically die between days 18-20. Positive compounds avert or delay the markers of mitochondrial dysfunction and avert or delay neuronal cell death, remarkable protective properties against the disease-causing neuropathology of ALS.

[0278] One compound screened, dipyridamole (DPM, Figure 3), also known as persantine, was remarkably effective in all parameters measured, including preservation of C9 MN well beyond their normal lifespan and with apparent mitochondrial health, by several measures, comparable to heathy I non-diseased isogenic control motor neurons (ISO MN). As stated above, test compounds, including DPM and analogs described below, were added only once to the neurons, at day 12. As shown in Figure 4A-4C, DPM (open gray circles) improved mitochondrial count (Figure 4A), mitochondrial length (Figure 4B), and neurite cumulative area (Figure 4C) (a measure of axon / dendrite [neurite] degeneration) out to at least 30 days. Only the parameter of mitochondrial length (center graph in Figure 4B) showed partial and not full protection by DPM. Some of the analogs of DPM described below are fully effective (indistinguishable from the ISO MN) even in this parameter. Moreover, protective effects, including 09 MN survival, can be extended to at least 80 days, for the improved analogs of DPM.

[0279] The combined data suggest that molecules of this class protect the MT in the human MN from the insults of the disease, which in turn, protect MN axons and dendrites from degeneration and the MN from dying. The effects on MT health are visually quite apparent, as shown in Figure 5A and Figure 5B. Effects of DPM on 09 MN viability are dose-dependent, as shown in Figure 6, with levels of 5 piM and above being effective.

[0280] DPM and related molecules of this class also improve mitochondrial function, as measured by mitochondrial respiration in 09 MN to that seen for ISO MN (Figure 7A-7D). In these studies, the Seahorse Cell Metabolic Analyzer was used to measure the oxygen consumption rate (OCR, or respiration) of the 09 and ISO MN. The OCR provides a measure of the ability of MT to use oxygen to synthesize chemical energy in the form of ATP as four different metrics: the basal OCR (Fig. 7A), the OCR related to ATP generation (Fig. 7B), the maximal OCR of the MT (Fig. 70), and the spare OCR of the MT (Fig. 7D). Molecules of this class when presented to the 09 MN increase the OCR across all four metrics such that they are indistinguishable from the ISO MN at a concentration as low as 39 nM.S-T00387W0001 32917 / 70090 / PC

[0184] The earliest studies of DPM pharmacokinetics (PK) and CNS permeability claimed that DPM does not cross the blood: brain barrier (BBB) (Mellinger and Bohorfoush et al., 1965, and refs therein). This claim was based on the old technique of fluorescence spectrum measurements of DPM in tissue homogenates and has been propagated on popular websites and many subsequent literature reviews that present DPM's PK properties. However, recent studies have shown that DPM does gain access to CNS tissue in an appreciable amount, depending upon its formulation and mode of administration. This has permitted the use of DPM in clinical trials for the CNS disorders of restless legs syndrome and schizophrenia (Garcia-Borreguero et al., 2018; 2021; Akhondzadeh et al., 2000; Lintunen et al., 2021). It also has efficacy in animal models for several CNS diseases (Sloka et al., 2013; Ferree et al. 2018, 2019; Oprisoreanu et al., 2021). Most importantly, DPM's brain penetration was measured directly using sensitive and modern techniques (HPLC-MS / MS) in a study on the breast cancer resistance protein (BCRP) in BBB function (Zhao et al., 2009), a study not part of the mainstream literature on DPM's BBB permeability. The brain / plasma concentration ratio of treated C57BI / 6J mice measured 0.25. Hervey and Goa (1999) measured plasma levels at 3.5 piM in humans administered DPM, which equates from the 0.25 ratio to a CNS concentration of approximately 0.9 piM, within the EC50 range measured for various metrics in C9 IPSC-derived MN (Figure 8; Table 1).Dipyridamole (DPM)Parameter ECso / Span Comments Activity for MT length* 1.5 piM / 46 Fig 8 Activity for MT count* 0.95 piM / 5.5 Fig 8 Activity for Neurite GA* 1.1 piM / 9 Fig 8 Activity for MN survival* 2-5 piM Fig 6 Activity for MT function >39 nM Fig 7Table 1. DPM Profile. *The following 4 measures were used: avg. MTlength, avg MT count, neurite (axons and dendrites) cumulative area (GA;e.g., complexity), and MN survival. EC50 and span values for C9-IPSC-derived MN at day 20 in D: R experiments.

[0281] With respect to DPM's structure and properties, it has a high molecular weight (505), high topological polar surface area (—143), and four hydrogen bond donors, parameters that one would prefer to moderate to gain high blood brain barrier permeability, low efflux liability, and prolonged exposure. It is reasonably water-soluble (cLogP ~1.5).

[0282] DPM analogs described are improved in biological and chemical properties. Figure 9 illustrates an example of an analog (DPM-A19) improving DPM's ability to protect against MT fragmentation in ALS C9orf72 IPSC derived motor neurons. Figure 10 illustrates the ability of a representative analog (DPM-A19) to improve the potency (EC50) and efficacy (Span) compared to DPM, in a side-by-side experiment comparing D: R properties at day 18. Note that there is some variability in these parameters between experiments (Figure 8 vs Figure 10) and as a function of day-in-culture (Day 20 for Figure 8 and Day 18 for Figure 10). Some of the analogs are predicted to have greater blood brain barrier permeability, with reduced hydrogen bond donors, lower topological polar surface area, and reduced molecular weight.S-T00387W0001 32917 / 70090 / PCEfficacy in AD iPSC-Derived Glutamatergic Cortical Neurons

[0283] The disclosed compounds were tested in AD iPSC-derived cortical neurons carrying presenilin mutations. Figures 11A-C illustrate data measuring MT count (Fig. 11 A), MT length (Fig. 11 B), and axon / dendrite integrity (Neurite CA) (Fig. 11C) across days 12-30 of culture. The representative compounds tested promote all three parameters relative to untreated AD iPSC-derived neurons, with potency of a representative compound (DPM-A19) between 0.7 and 1.8 μM. The compounds tested were DMSO (control), dipyridamole, and two of the DPM analogs (DPM-A7b, and DPM-A19). These data demonstrate that the compounds offset the loss of MT content, MT fragmentation, and axon / dendrite degeneration due to a mutation that causes AD. Each of the bottom panels of Figures 11 A-C illustrate the dose dependent effects of the compounds on these MT parameters. Figures 12A-D illustrate the dose:response effect of DPM on the impaired MT bioenergetics of IPSC-derived cortical neurons carrying the AD PS1-A246E mutation compared to the isogenic control CN. Increasing concentrations of DPM offset the impaired bioenergetics of the AD neurons with complete normalization of the four bioenergetics parameters by 2.5 μm DPM.Efficacy in TDP43-ALS Motor Neurons

[0284] The disclosed compounds were tested in iPSC-derived motor neurons (MN) carrying the TDP43 M337V mutation, a genetic form of ALS. Figure 13A-C (top) illustrates data measuring MT count, MT length, and axon / dendrite integrity (Neurite CA) across days 8-30 of culture for ALS-TDP43 MN carrying a M337V mutation and the control MN. DPM promotes all three parameters in the TDP43 mutant MN relative to untreated (DMSO) TDP43 mutant MN, so that the measured values are increased to levels similar to the control MN. Dose:response data (bottom) show that the potency is in the low micromolar range, with EC50 values of approximately 1.5 μM for MT count, 1.9 μM for MT length, and 1.3 μM for Neurite CA. These data demonstrate that the compounds offset the loss of MT content, MT fragmentation, and axon / dendrite degeneration due to the disease caused by the TDP43 mutation.Efficacy in SOD1-ALS Motor Neurons

[0285] The disclosed compounds were also tested in iPSC-derived motor neurons (MN) carrying the SOD1 A5V mutation, another genetic form of ALS. Figure 14A-C (top) illustrates data measuring MT count, MT length, and axon / dendrite integrity (Neurite CA) across days 8-30 of culture for ALS-SOD1 MN carrying an ALS-SOD1 A5V mutation and the control MN. DPM promotes all three parameters in the ALS-SOD1 A5V mutant MN relative to untreated (DMSO) ALS-SOD1 A5V mutant MN, so that the measured values are increased to levels similar or greater to the control MN. Dose:response data (bottom) show that the potency is in the low micromolar range, with EC50 values of approximately 3.7 μM for MT count, 2.8 μM for MT length, and 2.1 μM for Neurite CA. These data demonstrate that the compounds offset the loss of MT content, MT fragmentation, and axon / dendrite degeneration due to the disease caused by the mutations at the SOD1 gene.S-T00387W0001 32917 / 70090 / PCPromotion of Pyruvate Uptake into Mitochondria

[0286] The disclosed compounds were tested for their ability to promote pyruvate uptake into mitochondria. Figures 15 and 16 shows the results of pyruvate uptake experiments using isolated mouse liver mitochondria. UK5099, a known MPC inhibitor, demonstrated dose-dependent inhibition of pyruvate uptake, as expected. In contrast, DPM and a DPM analog A23 showed enhanced pyruvate uptake at various concentrations compared to baseline. These data demonstrate that the disclosed compounds work by promoting pyruvate uptake into mitochondria, in contrast to known MPC inhibitors which block pyruvate uptake.REFERENCESAggrenox Monograph. (2015). Boehringer Ingelheim Ltd, available online.Akhondzadeh S, Shasavand E, Jamilian H, Shabestari O, Kamalipour A. (2000). Dipyridamole in the treatment of schizophrenia: adenosine-dopamine receptor interactions. J Clin Pharm Ther. 25, 131-7.Allahham M, Lerman A, Atar D, Birnbaum Y. (2022). 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Claims

S-T00387W0001 32917 / 70090 / PCWHAT IS CLAIMED:

1. A compound of formula (I):or a pharmaceutically acceptable salt thereof,whereinA1is selected from morpholinyl substituted with 0-2 substituents independently selected from C^alkyl and Ci-3hydroxyalkyl; piperidinyl substituted with 0-2 substituents independently selected from C^alkyl, hydroxy, and Ci.c\zo oo N O N N3hydroxyalkyl; -N(R3)2, -J-, -~L~, and o ~L-;A2is selected from piperidinyl, pyrrolidinyl, azetidinyl, -N(R4)2, -O-Ci-shydroxyalkyl, -S-Ci-3hydroxyalkyl, halo, and morpholinyl substituted with 0-2 substituents independently selected from Ci-3alkyl, and Ci-3hydroxyalkyl, wherein the piperidinyl, and pyrrolidinyl are substituted with 0-3 substituents independently selected from Ci-3alkyl, Ci.3alkoxy, hydroxy, Ci-3hydroxyalkyl, phenyl, and halo;A1or A2optionally further comprise a labile linker;R1and R2are each independently N- or CH, wherein R1and R2together with carbon and nitrogen atoms to which they are attached form a 5- or 6-membered ring, having 0-2 ring N atoms, and the 5- or 6-membered ring is substituted with 0-2 substituents independently selected from A3and -Ci-3alkylene-O-Ci-3alkyl;each A3is independently selected from morpholinyl substituted with 0-2 substituents independently selected from Ci-3alkyl, and Ci-3hydroxyalkyl; piperidinyl, pyrrolidinyl, -N(R5)2, -Ci-3hydroxyalkyl, and -Ci-3alkylene-Ci-3alkoxy, wherein the piperidinyl and pyrrolidinyl are substituted with 0-2 substituents independently selected from Ci-3alkyl, hydroxy, and Ci-3hydroxyalkyl;each R3is independently C ealkyl or Ci-3hydroxyalkyl; andeach R4and R5is independently selected from H, Ci-ealkyl, Ci-ehydroxyalkyl, and Ci-3alkyl-O-Ci-6alkylene;with the proviso that the compound is notor and at least one of A1, A2, and A3is piperidinyl or pyrrolidinyl optionally substituted with 0-3 substituentsS-T00387W0001 32917 / 70090 / PCindependently selected from C^alkyl, C^alkoxy, hydroxy, Ci-shydroxyalkyl, phenyl, and halo, or at least one of A1, A2, and A3is morpholinyl optionally substituted with 0-2 substituents independently selected from Ci^alkyl and Ci- shydroxyalkyl, or at least one R4or R5is Ci-ehydroxyalkyl, and C1.3alkyl-O-C1.6al ky lene.

2. The compound or pharmaceutically acceptable salt of claim 1, wherein A1is selected from -J—,3. The compound or pharmaceutically acceptable salt of claim 1 or 2, wherein A2is selected from4. The compound or pharmaceutically acceptable salt of any one of claims 1-3, wherein at least one of R1and R2is C^alkyl.

5. The compound or pharmaceutically acceptable salt of any one of claims 1-4, having a formula:X1and X2are each independently N or CH.

6. The compound or pharmaceutically acceptable salt of claim 5, wherein at least one X1is N.

7. The compound or pharmaceutically acceptable salt of claim 5, wherein at least one X1is CH.

8. The compound or pharmaceutically acceptable salt of any one of claims 5-7, having a formulaS-T00387W0001 32917 / 70090 / PC9. The compound or pharmaceutically acceptable salt of any one of claims 1-4, having a formulawhereinX2is N or CH; andA4, A5, and A6are each independently H, Ci^alkyl, morpholinyl substituted with 0-2 substituents independently selected from Ci-3alkyl, and Ci-shydroxyalkyl, piperidinyl, pyrrolidinyl, -N(R5)2, Ci-shydroxyalkyl, and Ci-3alkylene-O-Ci-3alkyl, wherein the morpholinyl, piperidinyl and pyrrolidinyl are substituted with 0-2 substituents independently selected from Ci-salkyl, hydroxy, and Ci-shydroxyalkyl.

10. The compound or pharmaceutically acceptable salt of claim 9, wherein X2is N.

11. The compound or pharmaceutically acceptable salt of claim 9, wherein X2is CH.

12. The compound or pharmaceutically acceptable salt of any one of claims 9-11, wherein A4is H.

13. The compound or pharmaceutically acceptable salt of any one of claims 9-11, wherein A4is Ci. 4alkyl.

14. The compound or pharmaceutically acceptable salt of any one of claims 9-11, wherein A4is selected from morpholinyl substituted with 0-2 substituents independently selected from C salkyl and Ci- shydroxyalkyl, piperidinyl, pyrrolidinyl, -N(R5)2, -Ci-shydroxyalkyl, and -Ci-salkyl hydroxy-Ci-salkyl, wherein the morpholinyl, piperidinyl and pyrrolidinyl are substituted with 0-2 substituents independently selected from C salkyl, hydroxy, and Ci-shydroxyalkyl.

15. The compound or pharmaceutically acceptable salt of claim 14, wherein A4is selected from H, OH '-o, and,16. The compound or pharmaceutically acceptable salt of any one of claims 9-15, wherein A5is H.

17. The compound or pharmaceutically acceptable salt of any one of claims 9-15, wherein A5is Ci.S-T00387W0001 32917 / 70090 / PC4alkyl.

18. The compound or pharmaceutically acceptable salt of any one of claims 9-17, wherein A6is H.

19. The compound or pharmaceutically acceptable salt of any one of claims 9-17, wherein A6is Ci. 4alkyl.

20. The compound or pharmaceutically acceptable salt of any one of claims 1-19, wherein the labile linker comprises a functional group selected from a hydrazone, a disulfide, an ester, a carbamate, a peptide, an azo, and an oxime.

21. The compound or pharmaceutically acceptable salt of any one of claims 1 -20, wherein the labile linker comprises a phosphate or phosphonate ester, sulfate or sulfonic ester, or a carboxylic ester, wherein the ester comprises an alkyl or amino acid residue.

22. The compound or pharmaceutically acceptable salt of claim 1, selected fromS-T00387W0001 32917 / 70090 / PCS-T00387W0001 32917 / 70090 / PC DPM-A32, DPM-A33,S-T00387W0001 32917 / 70090 / PCS-T00387W0001 32917 / 70090 / PCDPM-A61.

23. The compound or pharmaceutically acceptable salt of claim 22, selected from 1,S-T00387W0001 32917 / 70090 / PCDPM-A20,S-T00387W0001 32917 / 70090 / PC24. The compound or pharmaceutically acceptable salt of claim 22, selected from DPM-A2, DPM-A6, DPM-A10, DPM-A11, DPM-A13, DPM-A17, DPM-A19, DPM-A20, DPM-A21, DPM-A22, DPM-A23, DPM-A24, DPM-A28, DPM-A36, DPM-A35, DPM-A34, DPM-A39, DPM-A40, DPM-A46, DPM-A47, DPM-A41, DPM-A42, DPM-A43, DPM-A44, DPM-A45, DPM-A48, DPM-A49, DPM-A50, DPM-A52, DPM-A54, and DPM-A55.

25. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the compound or pharmaceutically acceptable salt of any one of claims 1-24.

26. The pharmaceutical composition of claim 25, wherein the pharmaceutical composition is formulated for oral, parenteral, inhalational, intranasal, subcutaneous, intramuscular, or intravenous administration.

27. The pharmaceutical composition of claim 26, wherein the pharmaceutical composition is formulated for oral administration.

28. The pharmaceutical composition of any one of claims 25-27, further comprising an additional therapeutic agent.

29. A method of treating a neurodegenerative disease or disorder in a subject diagnosed or predisposed to: i. Parkinson's disease and related disorders including but not limited to Parkinson's disease, Parkinson-dementia, autosomal recessive PARK2 and PARK6-linked Parkinsonism, atypical parkinsonian syndromes, including, progressive supranuclear palsy, corticobasal degeneration syndrome, Lewy bodies dementia, multiple system atrophy, Guadeloupean Parkinsonism and Lytigo-bodig disease; ii. motor neuron diseases including but not limited to amyotrophic lateral sclerosis, frontotemporal dementia, progressive bulbar palsy, pseudobulbar palsy, primary lateral sclerosis, progressive muscular atrophy, spinal muscular atrophy, flail arm syndrome, flail leg syndrome, ALS-plus syndrome, and post-polio syndrome; ill. neuro-inflammatory diseases including but not limited to autoimmune encephalitis, optic neuritis, transverse myelitis, neurosarcoidosis, neuromyelitis optica, traumatic brainS-T00387W0001 32917 / 70090 / PCinjury, post concussive syndrome, multiple sclerosis, chronic traumatic encephalopathy, spinal cord injury; iv.Alzheimer's disease and related disorders including but not limited to early stage of an Alzheimer's disorder, mild stage of an Alzheimer's disorder, moderate stage of an Alzheimer's disorder, mild to moderate stage of an Alzheimer's disorder, advanced stage of an Alzheimer's disorder, mild cognitive impairment, vascular dementia, mixed dementia, Pick's disease, argyrophilic grain disease, posterior cortical atrophy, Wernicke-Korsakoff Syndrome; v. prion diseases including but not limited to Creutzfeldt-Jakob disease, Kuru, variant Creutzfeldt-Jakob disease, Gerstmann-Straussler-Scheinker Syndrome, fatal familial insomnia; vi. lysosomal storage diseases including but not limited to Fabry disease, mucopolysaccharidoses, Niemann-Pick disease, Tay-Sachs disease; vii. Leukodystrophies including but not limited to Alexander disease, Canavan disease, Krabbe disease, Adrenoleukodystrophy, Refsum disease; viii. Huntington's disease; ix. multiple sclerosis (MS) including but not limited to secondary progressive MS, progressive relapsing MS, benign MS, primary progressive MS; x. Down syndrome; xi. spinal and bulbar muscular atrophy (SMA) including but not limited to acute infantile SMA, chronic juvenile SMA, adult onset SMA; xii. HIV-Associated Neurocognitive Disorders including but not limited to HIV-associated dementia, mild neurocognitive disorder; xiii.Tourette Syndrome; xiv. spinocerebellar ataxia (SCA) including but not limited to dominant SCA1-35, autosomal recessive SCA, X-linked SCA; xv. Dentatorubral pallidoluysian atrophy; xvi. myotonic dystrophy; xvii, schizophrenia and schizoaffective disorder; xviii. Bipolar spectrum disorders; xix. autism and autism spectrum disorders; xx. attention-deficit hyperactivity spectrum disorders; xxi. chronic pain; xxii. alcohol-induced dementia; xxiii. progressive non-fluent aphasia; xxiv. semantic dementia; xxv. spastic paraplegia; xxvi. fibromyalgia; xxvii. post-Lyme disease; xxviii. neuropathies; xxix. withdrawal symptoms; xxx. Alpers' disease; xxxi. cerebro-oculo-facio-skeletal syndrome; xxxii. Wilson's disease; xxxiii. Cockayne syndrome; xxxiv. Leigh's disease and other primary mitochondrial diseases; xxxv. neurodegeneration with brain iron accumulation; xxxvi. opsoclonus myoclonus syndrome; xxxvii. alpha-methylacyl-CoA racemase deficiency; xxxviii. Andermann syndrome; xxxix. Arts syndrome; xxxx. Marinesco-Sjogren syndrome; xxxxi. mitochondrial membrane protein-associated neurodegeneration; xxxxii. pantothenate kinase-associated neurodegeneration; xxxxiii. polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy; xxxxiv. riboflavin transporter deficiency neuronopathy; xxxxv. ataxia telangiectasial; xxxxvi. age associated memory impairment; and Batten disease by administering a therapeutically effective amount of the compound or pharmaceutically acceptable salt of any one of claims 1 -24 or the pharmaceutical composition of any one of claims 25-28.

30. The method of claim 29, wherein the neurodegenerative disease or disorder is selected from Amyotrophic Lateral Sclerosis (ALS), Alzheimer’s disease (AD), Lewy body dementia (LBD), Parkinson’s disease (PD), Huntington’s disease, spinal muscular atrophy (SMA), primary lateral sclerosis (PLS), progressive muscular atrophy (PMA), Kennedy's disease (spinal and bulbar muscular atrophy), post-polio syndrome, motor neuron disease (MND), multiple sclerosis (MS), multifocal motor neuropathy (MMN), hereditary spastic paraplegia (HSP), myasthenia gravis, and Friedreich's ataxia.S-T00387W0001 32917 / 70090 / PC31. The method of claim 30, wherein the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), or Parkinson’s disease (PD).

32. The method of any one of claims 29-31, wherein the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), progressive muscular atrophy, primary lateral sclerosis, progressive bulbar palsy, flail arm syndrome, flail leg syndrome, and ALS-plus syndrome.

33. The compound or pharmaceutically acceptable salt of any one of claims 1 -24 or the pharmaceutical composition of any one of claims 25-28, for use in treating a neurodegenerative disease or disorder.

34. The method of claim 29, wherein the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS) associated with a mutation in the C9orf72 gene.

35. The method of claim 34, wherein the C9orf72 mutation is a hexanucleotide repeat expansion.

36. The method of claim 29, wherein the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS) associated with a mutation in the TDP43 gene.

37. The method of claim 36, wherein the TDP43 mutation is M337V.

38. The method of claim 29, wherein the neurodegenerative disease or disorder is amyotrophic lateral sclerosis (ALS) associated with a mutation in the SOD1 gene.

39. The method of claim 38, wherein the SOD1 mutation is A5V.

40. The method of claim 29, wherein the neurodegenerative disease or disorder is sporadic ALS.

41. The method of claim 29, wherein the neurodegenerative disease or disorder is Alzheimer's disease (AD).

42. The method of claim 41, wherein the Alzheimer's disease is associated with a mutation in the presenilin 1 (PS1) gene or presenilin 2 (PS2) gene.

43. The method of claim 41, wherein the Alzheimer's disease is sporadic Alzheimer's disease.

44. A method of promoting pyruvate uptake into mitochondria in a subject in need thereof, comprising administering a therapeutically effective amount of the compound or pharmaceutically acceptable salt of any one of claims 1-24 or the pharmaceutical composition of any one of claims 25-28.

45. The method of claim 44, wherein the compound enhances pyruvate transport across the mitochondrial membrane.

46. The method of claim 44 or 45, wherein pyruvate transport enhancement is mediated by Mitochondrial Pyruvate Carrier (MPC).S-T00387W0001 32917 / 70090 / PC47. A method of treating a disease or disorder characterized by impaired mitochondrial pyruvate uptake, comprising administering a therapeutically effective amount of the compound or pharmaceutically acceptable salt of any one of claims 1 -24 or the pharmaceutical composition of any one of claims 25-28.

48. A method of preserving mitochondrial health in motor neurons of a subject having a neurodegenerative disease, comprising administering a therapeutically effective amount of the compound or pharmaceutically acceptable salt of any one of claims 1-24, wherein the method results in one or more of: (a) increased mitochondrial count; (b) increased mitochondrial length; (c) increase mitochondrial bioenergetics; (d) increased neurite cumulative area; and (e) extended motor neuron viability.

49. The method of claim 47 or 48, wherein the neurodegenerative disease is selected from ALS and Alzheimer's disease.

50. The method of claim 47 or 48, wherein the neurons are selected from motor neurons and cortical neurons.

51. The method of claim 47 or 48, wherein the ALS is selected from C9orf72-ALS, SOD1-ALS, TDP43-ALS, other fALS forms, and sporadic ALS.

52. The method of claim 47 or 48, wherein the Alzheimer's disease is associated with a mutation in the presenilin 1 (PS1) gene or presenilin 2 (PS2) gene, APP gene, other fAD forms, and sporadic AD.

53. The method of any one of claims 29-52, wherein the compound increases at least one parameter selected from mitochondrial count, mitochondrial length, mitochondrial bioenergetics, neurite cumulative area, and neuronal survival to levels similar to those observed in non-diseased control neurons.

54. The method of claim 53, wherein the compound increases mitochondrial count, mitochondrial length, and neurite cumulative area.

55. The method of any one of claims 29-54, wherein the EC50for one or more of mitochondrial count, mitochondrial length, and neurite cumulative area is in the low micromolar range.

56. The method of claim 55, wherein the EC50is from about 0.4 μM to about 4 μM.

57. The method of claim 55, wherein the EC50 is from about 0.7 μM to about 4 μM.

58. The method of claim 53, wherein the compound increases mitochondrial bioenergetics.

59. The method of any one of claims 29-58, wherein the EC50for mitochondrial bioenergetics is about 0.04 piM to about 2.2 piM.

60. The method of any one of claims 29-59, wherein the compound is dipyridamole (DPM) or a structural analog thereof.S-T00387W0001 32917 / 70090 / PC61. The method of claim 60, wherein the compound is dipyridamole (DPM).

62. The method of claim 54, wherein the compound is compound 23 (DPM-A23) of formula

Images