Deuterated hinokiol derivative

Abstract

Deuterated derivatives of tropolone useful in the treatment of diseases or disorders, such as diseases or disorders involving iron transport defects, are disclosed.

Description

[0001] Related Applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 411,738, filed on September 30, 2022.

[0003] Government funding statement

[0004] This invention was made with government support under Grant 1R01HL140526 awarded by the National Institutes of Health. The government has certain rights in this invention. Background Art

[0005] Hinokitiol, a small molecule natural product, has been shown to promote transmembrane iron transport by reversibly forming a 3:1 complex with iron. Hinokitiol restores iron homeostasis in a range of different model systems, including divalent metal transporter 1 (DMT1-), mitochondrial protein 1 (MFRN1-), and ferroportin 1 (FPN1-) deficient cells and animals. Hinokitiol, which is not site- or direction-selective per se, achieves site- and direction-selective iron transport by exploiting the electrochemical gradient formed in the absence of ferroportin function.

[0006] However, hinokitiol has a key limitation that diminishes its potential for clinical translation. In differentiated shDMT1 monolayers, hinokitiol has been found to effectively mobilize iron at low to moderate concentrations, but at higher concentrations, a significant reduction in transmembrane migration activity was observed. A series of studies were conducted to identify derivatives of hinokitiol, such as those disclosed herein, that mobilize iron over a wider range of concentrations in vitro. These compounds are also able to mobilize iron and may treat anemia of inflammation in vivo.

[0007] During the preparation of these improved derivatives, it was identified that at higher concentrations of hinokitiol, the loss of iron transport led to the formation of higher order aggregates in aqueous media. These aggregates then limited the amount of free tropolone: ​​iron complexes that were able to transport iron across the membrane. In order to understand how these complexes interact in these aggregates, several crystal structures of hinokitiol: M (M = Fe, Al, Cu) complexes were obtained. In these crystal structures, the isopropyl groups of these complexes formed key van der Waals interactions that may drive the formation of aggregates in aqueous media. Using constitutional isomers of hinokitiol that are less prone to aggregation, the van der Waals interactions generated by the isopropyl groups were significantly different.

[0008] Deuterium is an isotope of hydrogen, with a nucleus containing one neutron, doubling the atomic weight. This reduces the zero-point energy and van der Waals radius of deuterated compounds relative to their hydrogenated counterparts. However, the literature on the effects of deuteration in drugs has mainly focused on exploiting the kinetic isotope effect to improve the pharmacokinetic properties of drugs. Some studies on deuterated materials have reported changes in boiling points, dimerization behavior, etc. There have been few systematic studies to explain the relationship between deuteration and the altered biophysical behavior of pharmaceuticals. Thus, it is unclear whether deuteration has any effect on such aggregation phenomena, nor whether any effect is positive or negative. Using substituted tropolones, it has been found that deuteration of the side chain of tropolone-type compounds can increase potency and tolerability, thus enhancing their potential as molecular prosthetics for metal ion transporters. Summary of the Invention

[0009] In certain aspects, the present disclosure provides compounds of Formula I:

[0010]

[0011] or a pharmaceutically acceptable salt thereof;

[0012] Wherein:

[0013] R 1 is selected from the group consisting of: H, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl, and -O-heterocycloalkyl;

[0014] R 2 is selected from the group consisting of: H, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl, and -O-heterocycloalkyl; and

[0015] R 3 is selected from the group consisting of: H, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl, and -O-heterocycloalkyl;

[0016] R 4 is selected from the group consisting of: H, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl, and -O-heterocycloalkyl; and

[0017] R 5 is selected from the group consisting of: H, halo, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl, and -O-heterocycloalkyl;

[0018] Wherein when R 1 、R 2 、R 3 、R4 or R 5 When it is C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl or -O-heterocycloalkyl, at least one hydrogen atom in the C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl or -O-heterocycloalkyl is replaced by a deuterium atom; and

[0019] wherein R 1 , R 2 , R 3 and R 4 at least one of them is not H.

[0020] In a further aspect, the present disclosure provides a pharmaceutical composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient.

[0021] In yet a further aspect, the present disclosure provides a method for treating a disease or disorder characterized by reduced ferroportin, which comprises administering to a subject in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure.

[0022] In yet a further aspect, the present disclosure provides a method for treating a disease or disorder characterized by reduced ferrotransferrin, which comprises administering to a subject in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure.

[0023] In certain aspects, the present disclosure provides a method for treating a disease or disorder characterized by ferrotransferrin deficiency or insufficiency, which comprises administering to a subject in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure.

[0024] In a further aspect, the present disclosure provides a method for treating a disease or disorder characterized by iron accumulation, which comprises administering to a subject in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure.

[0025] In yet a further aspect, the present disclosure provides a method for treating a disease or disorder characterized by improper iron distribution, which comprises administering to a subject in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure.

[0026] In certain aspects, the present disclosure provides a method for treating neurodegenerative diseases, which comprises administering to a subject in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Depicts a general synthetic scheme for obtaining deuterated tropolone derivatives.

[0028] Figure 2ADepicts data evaluating methyl-d0 and methyl-d3 tropolone. Iron mobilized by the deuterated derivative increases at low concentrations and remains at high concentrations at extreme concentrations.

[0029] Figure 2B Depicts data evaluating methyl-d0 and methyl-d3 tropolone. The deuterated derivative is significantly better tolerated in H9C2 cells.

[0030] Figure 2C Depicts data evaluating methyl-d0 and methyl-d3 tropolone. The deuterated derivative is better tolerated in K562 cells.

[0031] Figure 3A Depicts the results of treating fpn-1.2KO Caenorhabditis elegans with various compounds for neurodegeneration scoring and iron level measurement: fpn-1.2KO treated with different concentrations of DFP, hinokitiol, and AMB-1269; treatment protocol for Pdat::GFP worms. Dopaminergic neurodegeneration was blindly scored by phenotypic analysis (as follows).

[0032] Figure 3B Depicts the dopaminergic neurodegeneration score of fpn-1.2KO worms treated with DFP.

[0033] Figure 3C Depicts the dopaminergic neurodegeneration score of fpn-1.2KO worms treated with hinokitiol.

[0034] Figure 3D Depicts the dopaminergic neurodegeneration score of fpn-1.2KO worms treated with FeM-1269.

[0035] Figure 3E Depicts the treatment protocol for fpn-1.2KO; Pftn-1::GFP worms treated with different concentrations of DFP, hinokitiol, and FeM-1269. ASI neurons expressing the GFP-tagged ferritin level (below).

[0036] Figure 3F Depicts the fluorescence level of fpn-1.2KO worms treated with DFP. By one-way ANOVA, *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001.

[0037] Figure 3G Depicts the fluorescence level of fpn-1.2KO worms treated with Hino. By one-way ANOVA, *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001.

[0038] Figure 3H Depicts the fluorescence levels of fpn-1.2KO worms treated with FeM-1269. By one-way ANOVA, *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001.

[0039] Figure 4A Depicts data from experiments on iron-deficient mice: iron-deficient mice exhibited increased anxiety and decreased exploratory activity. WT and iron-deficient mice were subjected to the elevated plus maze task to evaluate anxiety-like behaviors, including the total distance traveled and average speed throughout the maze.

[0040] Figure 4B Depicts data from experiments on iron-deficient mice: iron-deficient mice exhibited increased anxiety and decreased exploratory activity. WT and iron-deficient mice were subjected to the elevated plus maze task to evaluate anxiety-like behaviors, including the time spent in the open arms and central area.

[0041] Figure 4C Depicts data from experiments on iron-deficient mice: iron-deficient mice exhibited increased anxiety and decreased exploratory activity. WT and iron-deficient mice were subjected to the elevated plus maze task to evaluate anxiety-like behaviors, including the frequency and duration of rearing. By student t-test, *P<0.05.

[0042] Figure 4D Depicts ICP-MS measurements of brain iron levels in iron-deficient mice after acute treatment with hinokitiol by intraperitoneal (IP) injection. By one-way ANOVA, **P<0.01, ***P<0.001.

[0043] Figure 4E Depicts ICP-MS measurements of brain iron levels in iron-deficient mice after long-term treatment with hinokitiol by intraperitoneal (IP) injection. By one-way ANOVA, **P<0.01, ***P<0.001.

[0044] Figure 5 Depicts crystallographic data showing the formation of a 3:1 complex of hinokitiol with Fe(III) for transmembrane migration. The hinokitiol in the crystal packing structure contains isopropyl side chains adjacent to the complex.

[0045] Figure 6 Depicts a schematic diagram showing that deuteration is hypothesized to reduce van der Waals interactions between complexes, thereby restoring the transport ability of pre-aggregated derivatives at higher concentrations.

[0046] Figure 7A Depicts cell-free iron efflux data showing that full deuteration of hinokitiol promotes transport at higher concentrations, similar to that observed with 3,5,7-trimethylcycloheptatrienone.

[0047] Figure 7B Depicts dynamic light scattering data showing the reduced aggregation of fully deuterated hinokitiol.

[0048] Figure 8A Depicts crystallization data of hinokitiol showing isopropyl-isopropyl interactions.

[0049] Figure 8B Depicts cell-free iron efflux data showing that the full deuteration of the isopropyl isomer GT does not affect transport at higher concentrations, which is consistent with the limited specific isopropyl-isopropyl interactions observed in the crystal structure.

[0050] Figure 9A Depicts dynamic light scattering data (deflection count) showing that the full deuteration of the methyl side chain does not significantly affect aggregation due to the lack of discrete interactions and the limited aggregation of the parent compound.

[0051] Figure 9B Depicts dynamic light scattering data (average size) showing that the full deuteration of the methyl side chain does not significantly affect aggregation due to the lack of discrete interactions and the limited aggregation of the parent compound.

[0052] Figure 10A Depicts cell-free iron efflux data showing similar iron migration capabilities between 3-methyltropolone and 3-methyl-(d3)-tropolone.

[0053] Figure 10B Depicts cell-free iron efflux data showing similar iron migration capabilities between 4-methyltropolone and 4-methyl-(d3)-tropolone.

[0054] Figure 10C Depicts cell-free iron efflux data showing similar iron migration capabilities between 5-methyltropolone and 5-methyl-(d3)-tropolone.

[0055] Figure 10D Depicts cell-free iron efflux data showing similar iron migration capabilities between 3,5-dimethyltropolone and 3,5-dimethyl-(d6)-tropolone.

[0056] Figure 11 Depicts cell-free iron efflux data showing that compared to 4-n-butyltropolone, the full deuteration extends the transport window of pre-aggregated 4-n-butyl-(d9)-tropolone. Detailed Description

[0057] Iron is essential for organisms but also potentially toxic; therefore, it is tightly controlled at the cellular and systemic levels to prevent deficiency and overload. Iron overload is mainly manifested in two aspects: (1) excessive iron in circulation, and (2) iron accumulation in tissues. The latter can also be referred to as improper or uneven iron distribution. Iron chelators treat iron overload and enhance iron secretion by binding to iron in the blood and reducing the amount of iron in circulation. Iron mobilizers extract iron from tissues and thus can redistribute iron from the accumulation sites to the deficient sites. Such mobilizers can cooperate with ferritin to restore normal iron physiology. Current treatments mainly target excessive iron in the blood.

[0058] The transport of iron throughout the living system is achieved through the concerted activities of passive and active transport proteins. Disruption of iron homeostasis can be caused by diseases resulting from the malfunction of one of these proteins. Thujaplicin (β-thujaplicin), a natural product, autonomously substitutes for the function of ion transport proteins in disease models by forming a lipophilic 3:1 complex with iron and utilizing the transmembrane concentration gradient to redistribute iron. However, thujaplicin is prone to form higher-order aggregates at higher concentrations, thus limiting the effective range of mobilization. From the X-ray crystal structure of the complex, the β-substituted isopropyl groups of adjacent complexes promote aggregation through proximity-driven attractive van der Waals interactions. As observed in the synthetic design principles of new tropolone derivatives, reducing these interactions while maintaining sufficient lipophilicity for transmembrane mobilization will generally increase the range of mobilization efficacy. Notably, these new derivatives are generally structurally different from the aggregating compounds, and there is no alternative modification method to transform the aggregating compounds into compounds with reduced aggregation tendency but maintaining the same structural design.

[0059] Deuteration of small molecules is commonly used to alter pharmacokinetics due to the kinetic isotope effect (KIE). When it comes to the impact of deuteration on intermolecular van der Waals-type interactions, the literature is sparse and conflicting. Additionally, while normal primary KIE can result in ratio values of 1 to 8, the non-covalent deuterium isotope effect has been reported to be very modest, in the range of 1 - 1.5. Thus, it is unclear whether deuteration helps to address the aggregation-based loss of activity observed in tropolones such as hinokitiol, preliminary evidence suggesting that this is driven by higher-order applications. However, the sparse applications in supramolecular host-guest interactions, rotational barriers, chromatography, and self-association are intriguing and suggest to us that physicochemical differences that impact aggregation, albeit slightly, may be achievable through deuteration, especially in the case of fully deuterated species where these individual effects may be amplified. Therefore, despite strong literature precedent, we decided to experimentally test whether deuteration of aggregation-prone small molecule metalophores could improve the availability of the higher concentration active transport complex by disrupting complex association. We specifically tested whether full deuteration of a pre-aggregating side chain could largely maintain its lipophilicity while increasing the in vitro activity range compared to the parent compound, due to the potential effect of deuteration disrupting complex van der Waals interactions, thereby reducing higher-order aggregation of the small molecule with minimal structural perturbation.

[0060] To first test the application of deuteration in iron mobilizers, hinokitiol-d7 was synthesized via a Negishi coupling of 2-iodopropane-d7 with a modular bromotropolone scaffold. Hinokitiol was used as a highly aggregated standard, and hinokitiol-d7 is the comparative isotopologue containing a fully deuterated isopropyl side chain. In the Fe3+ liposome efflux assay, hinokitiol-d7 had significantly greater iron transport capacity at higher concentrations (50 - 150 μM) when compared to hinokitiol, which begins to aggregate. This transport capacity was similar to that of the non-isopropyl structural isomer control trimethyl, which aggregates less but has the same cLogP as hinokitiol, indicating that hinokitiol-d7 also aggregates to a lesser extent. This high-concentration transport capacity level is not comparable to most other derivatives, which is an interesting development for the development of improved iron mobilizers. The lesser aggregation is supported by dynamic light scattering studies, which showed that 20 - 100 μM of hinokitiol-d7 with equimolar iron aggregates had less photon deflection counts and particle size aggregation. Interestingly, the isopropyl structural isomer GT (γ-thujaplicin), which was previously shown to have less aggregation than hinokitiol (and different crystal packing that limits the proximity of the isopropyl groups between complexes), did not show an effect of deuteration on cell-free iron transport. This indicates that deuteration of non-interacting side chains has no effect on transmembrane iron transport. This is supported by non-interacting methyl and methyl-d3 variants at all ring positions, which did not show an operational difference in cell-free iron transport and all had a low aggregation propensity. These methyl variants also did not show a significant difference in the shDMT1-Caco-2 monolayer transport experiment that was able to be run, indicating that the cell-free trend extends to biological systems. Multisubstituted derivatives such as 3’5DiMe also did not show any effect of deuteration, even with an increased number of deuterium substitutions, because the parent compound is essentially non-aggregating.

[0061] To support this differential deuteration effect that depends on the aggregation tendency of the parent compound, highly aggregated compound 4nBu and its fully deuterated isotopologue pair were synthesized and tested in a liposome efflux assay. Due to higher-order aggregation, 4nBu has a shortened effective transmembrane migration concentration range, thus halting significant transport at 40 μM of the compound. When the parent compound undergoes restricted higher-order aggregation, 4nBu-d9 (similar to hinokitiol-d7) maintains the transport level at a certain concentration. Notably, in this case, this range extends to 75 μM of the compound, which exhibits significant transmembrane iron transport before dropping to baseline, which is indicative that deuteration can be strategically used as a new design principle to extend the effective concentration range of aggregated tropolones. In certain embodiments, further primary alkyl derivatives can be synthesized using commercially available fully deuterated halides, which are converted to the corresponding boronic acids by Grignard-promoted trimethylborane addition and then used for Suzuki-Miyaura cross-coupling with a bromotropolone scaffold.

[0062] This disclosure is based on the surprising discovery that deuteration of the side chain of tropolone-type compounds can increase potency and tolerability, thereby enhancing their potential as molecular prosthetics for metal ion transporters. The compounds of the present disclosure can be used to treat diseases caused by improper iron distribution.

[0063] Definitions

[0064] As used herein, the term "organic moiety" refers to a monovalent group containing one or more carbon atoms. The organic moiety can be aromatic or can be derived from a hydrocarbon. The organic moiety can contain one or more heteroatoms, one or more unsaturated units, and / or one or more functional groups. The organic moiety can be substituted.

[0065] As used herein, the term "alkyl" is a technical term and refers to saturated aliphatic groups, including straight-chain alkyl, branched-chain alkyl, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl, and cycloalkyl-substituted alkyl. In certain embodiments, the straight-chain or branched-chain alkyl has about 30 or fewer carbon atoms in its main chain (e.g., for straight-chain, C1-C 30 , and for branched-chain, C3-C 30 ), and alternatively, about 20 or fewer or 10 or fewer carbon atoms. In certain embodiments, the term "alkyl" refers to C1-C 10 alkyl. In certain embodiments, the term "alkyl" refers to C1-C6 alkyl, such as C1-C6 straight-chain alkyl. In certain embodiments, the term "alkyl" refers to C3-C 12Branched alkyl. In certain embodiments, the term "alkyl" refers to a C3-C8 branched alkyl. Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, and n-hexyl.

[0066] The term "cycloalkyl" means a monocyclic or bicyclic or bridged saturated carbon ring, each having 3 to 12 carbon atoms. Some cycloalkyls have 5-12 carbon atoms in their ring structure and can have 6-10 carbon atoms in the ring structure. Preferably, the cycloalkyl is a (C3-C7) cycloalkyl, which represents a monocyclic saturated carbon ring having 3 to 7 carbon atoms. Examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. Bicyclic cycloalkyl ring systems include bridged monocyclic and fused bicyclic. The bridged monocyclic ring contains a monocyclic cycloalkyl ring, wherein two non-adjacent carbon atoms of the monocyclic ring are connected by an alkylene bridge of one to three additional carbon atoms (i.e., -(CH2) w - form of the bridging group, where w is 1, 2, or 3). Representative examples of bicyclic systems include, but are not limited to, bicyclo[3.1.1]heptane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, bicyclo[3.3.1]nonane, and bicyclo[4.2.1]nonane. The fused bicyclic cycloalkyl ring system contains a monocyclic cycloalkyl ring fused to a phenyl, monocyclic cycloalkyl, monocyclic cycloalkenyl, monocyclic heterocyclic, or monocyclic heteroaryl. The bridged or fused bicyclic cycloalkyl is attached to the parent molecular moiety through any carbon atom contained in the monocyclic cycloalkyl ring. The cycloalkyl is optionally substituted. In certain embodiments, the fused bicyclic cycloalkyl is a 5- or 6-membered monocyclic cycloalkyl ring fused to a benzene ring, 5- or 6-membered monocyclic cycloalkyl, 5- or 6-membered monocyclic cycloalkenyl, 5- or 6-membered monocyclic heterocyclic, or 5- or 6-membered monocyclic heteroaryl, wherein the fused bicyclic cycloalkyl is optionally substituted.

[0067] As used herein, the term "(cycloalkyl)alkyl" refers to an alkyl substituted with one or more cycloalkyls. An example of (cycloalkyl)alkyl is cyclohexylmethyl.

[0068] As used herein, the term "heterocycloalkyl" refers to a group that is a non-aromatic ring system, including but not limited to monocyclic, bicyclic, and tricyclic rings, which may be fully saturated or may contain one or more unsaturated units. For the avoidance of doubt, the degree of unsaturation does not result in an aromatic ring system, and it has 3 to 12 atoms, including at least one heteroatom such as nitrogen, oxygen, or sulfur. For purposes of illustration, and not to be construed as limiting the scope of the invention, examples of heterocycles are: aziridinyl, azirinyl, oxiranyl, thiiranyl, thiirenyl, dioxiranyl, diazirinyl, diazepanyl, 1,3-dioxolanyl, 1,3-dioxolyl, 1,3-dithiolanyl, 1,3-dithianyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, azetyl, oxetanyl, oxetyl, thietanyl, thietyl, diazetidinyl, dioxetanyl, dioxetenyl, dithietanyl, dithietyl, dioxalanyl, oxazolyl, thiazolyl, triazinyl, isothiazolyl, isoxazolyl, azepine, azetidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidinyl, oxopiperidinyl, oxopyrrolidinyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, quinuclidinyl, thiomorpholinyl, tetrahydropyranyl, tetrahydrofuranyl, tetrahydrothienyl, thiadiazolinyl, thiadiazolidinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, 1,1-dioxothiomorpholinyl (thiomorpholine sulfone), thiopyranyl, and trithianyl. The heterocycloalkyl is optionally substituted with one or more of the substituents described below.

[0069] As used herein, the term "(heterocycloalkyl)alkyl" refers to an alkyl group that is substituted with one or more heterocycloalkyl groups (i.e., heterocyclic groups).

[0070] As used herein, the term "alkenyl" means a straight or branched chain hydrocarbon group having 2 to 10 carbons and containing at least one carbon-carbon double bond formed by the removal of two hydrogens. Representative examples of alkenyl include, but are not limited to, vinyl, 2-propenyl, 2-methyl-2-propenyl, 3-butenyl, 4-pentenyl, 5-hexenyl, 2-heptenyl, 2-methyl-1-heptenyl, and 3-decenyl. The unsaturated bond of the alkenyl can be located at any position in the moiety and can have a (Z) or (E) configuration around the double bond.

[0071] As used herein, the term "alkynyl" means a straight or branched chain hydrocarbon group having 2 to 10 carbon atoms and containing at least one carbon-carbon triple bond. Representative examples of alkynyl include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 3-butynyl, 2-pentynyl, and 1-butynyl.

[0072] The term "alkylene" is well recognized in the art and, as used herein, refers to a diradical obtained by removing two hydrogen atoms of an alkyl group as defined above. In one embodiment, alkylene refers to a disubstituted alkane, i.e., an alkane substituted at two positions with substituents such as: halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, alkoxy, amino, nitro, mercapto, imino, acylamino, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, fluoroalkyl (such as trifluoromethyl), cyano, etc. That is, in one embodiment, "substituted alkyl" is "alkylene".

[0073] The term "amino" is a technical term and, as used herein, refers to unsubstituted and substituted amines, for example, a moiety that can be represented by the following general formula:

[0074]

[0075] wherein R a , R b and R c each independently represent hydrogen, alkyl, alkenyl, -(CH2) x -R d , or R a and R b together with the N atom to which they are attached form a heterocycle having 4 to 8 atoms in the ring structure; R d represents aryl, cycloalkyl, cycloalkenyl, heterocyclic group, or polycyclic group; and x is zero or an integer in the range of 1 to 8. In certain embodiments, only one of R a or R b can be a carbonyl, for example, R a , R bdoes not form an imide with nitrogen. In other embodiments, R a and R b (and optionally R c ) each independently represent hydrogen, alkyl, alkenyl or -(CH2) x -R d . In certain embodiments, the term "amino" means -NH2.

[0076] In certain embodiments, the term "alkylamino" means -NH(alkyl).

[0077] In certain embodiments, the term "dialkylamino" means -N(alkyl)2.

[0078] As used herein, the term "amido" means -NHC(=O), where the amido is bonded to the parent molecular moiety through nitrogen. Examples of amidos include alkylamidos such as CH3C(=O)N(H)- and CH3CH2C(=O)N(H)-.

[0079] The term "acyl" is a term of art as used herein and refers to any group or radical of the form RCO-, where R is any organic group such as alkyl, aryl, heteroaryl, aralkyl and heteroaralkyl. Representative acyls include acetyl, benzoyl and malonyl.

[0080] As used herein, the term "aminoalkyl" means an alkyl substituted with one or more amino groups. In one embodiment, the term "aminoalkyl" means aminomethyl.

[0081] The term "aminoacyl" is a term of art and as used herein refers to an acyl substituted with one or more amino groups.

[0082] As used herein, the term "aminothioyl" refers to an analogue of aminoacyl in which the O of RC(O)- is replaced by sulfur and thus has the form RC(S)-.

[0083] The term "phosphoryl" is a term of art and as used herein can generally be represented by the following formula:

[0084]

[0085] where Q50 represents S or O, and R59 represents hydrogen, lower alkyl or aryl; for example, -P(O)(OMe)- or -P(O)(OH)2. When used to substitute for example an alkyl, the phosphoryl of phosphorylalkyl can be represented by the following general formula:

[0086]

[0087] Wherein Q50 and R59 are each independently defined as above, and Q51 represents O, S or N; for example, -O-P(O)(OH)OMe or -NH-P(O)(OH)2. When Q50 is S, the phosphoryl moiety is a "thiophosphate ester".

[0088] As used herein, the term "aminophosphoryl" refers to a phosphoryl group substituted with at least one amino group, as defined herein; for example, -P(O)(OH)NMe2.

[0089] As used herein, the term "azide" or "azido" means an -N3 group.

[0090] As used herein, the term "carbonyl" refers to -C(=O)-.

[0091] As used herein, the term "thiocarbonyl" refers to -C(=S)-.

[0092] As used herein, the term "alkylphosphoryl" refers to a phosphoryl group substituted with at least one alkyl group, as defined herein; for example, -P(O)(OH)Me.

[0093] As used herein, the term "alkylthio" refers to alkyl-S-. The term "(alkylthio)alkyl" refers to an alkyl group substituted with an alkylthio group.

[0094] As used herein, the term "carboxyl" means a -CO2H group.

[0095] The term "aryl" is a technical term and, as used herein, refers to monocyclic, bicyclic, and polycyclic aromatic hydrocarbon groups, such as benzene, naphthalene, anthracene, and pyrene. Generally, an aryl group contains 6-10 carbon ring atoms (i.e., (C6-C 10 ) aryl). The aromatic ring may be substituted at one or more ring positions with one or more substituents such as: halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, alkoxy, amino, nitro, mercapto, imino, acylamino, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, fluoroalkyl (such as trifluoromethyl), cyano, etc. The term "aryl" also includes polycyclic ring systems having two or more rings, wherein two or more carbons are common to two adjacent rings (the rings are "fused rings"), wherein at least one ring is, for example, an aromatic hydrocarbon, and other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic group. In certain embodiments, the term "aryl" refers to phenyl.

[0096] The term "arylene" means a divalent group obtained by removing two hydrogen atoms of an aryl group as defined above. In certain embodiments, arylene refers to a disubstituted arene, i.e., an arene substituted at two positions with substituents such as: halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, alkoxy, amino, nitro, mercapto, imino, acylamino, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, fluoroalkyl (such as trifluoromethyl), cyano, etc. That is, in certain embodiments, "substituted aryl" is "arylene".

[0097] The term "heteroaryl" is a technical term and as used herein refers to monocyclic, bicyclic and polycyclic aryl groups having a total of 3 to 12 atoms and including one or more heteroatoms such as nitrogen, oxygen or sulfur in the ring structure. Exemplary heteroaryls include azaindolyl, benzo(b)thienyl, benzimidazolyl, benzofuranyl, benzoxazolyl, benzothiadiazolyl, benzotriazolyl, benzoxadiazolyl, furanyl, imidazolyl, imidazopyridyl, indolyl, indolinyl, indazolyl, isoindolinyl, isoxazolyl, isothiazolyl, isoquinolinyl, oxadiazolyl, oxazolyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridyl, pyrimidinyl, pyrrolyl, pyrrolo[2,3-d]pyrimidinyl, pyrrolo[3,4-d]pyrimidinyl, quinolinyl, quinazolinyl, triazolyl, thiazolyl, phenylthio, tetrahydroindolyl, tetrazolyl, thiadiazole, thienyl, thiomorpholinyl, triazolyl or tropanyl, etc. "Heteroaryl" may be substituted at one or more ring positions with one or more substituents such as: halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, alkoxy, amino, nitro, mercapto, imino, acylamino, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, fluoroalkyl (such as trifluoromethyl), cyano, etc. The term "heteroaryl" also includes polycyclic systems having two or more rings, wherein two or more carbons are common to two adjacent rings (the rings are "fused rings"), wherein at least one ring is an aromatic group having one or more heteroatoms in the ring structure, e.g., the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclic group.

[0098] The term "heteroarylene" means a divalent group obtained by removing two hydrogen atoms from a heteroaryl as defined above. In certain embodiments, heteroarylene refers to a disubstituted heteroarene, i.e., a heteroarene substituted at two positions with substituents such as: halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, alkoxy, amino, nitro, mercapto, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, fluoroalkyl (such as trifluoromethyl), cyano, etc. That is, in certain embodiments, "substituted heteroaryl" is "heteroarylene".

[0099] The term "aralkyl" or "arylalkyl" is a technical term and as used herein refers to an alkyl substituted with an aryl, wherein the moiety is attached to the parent molecule through the alkyl.

[0100] The term "heteroaralkyl" or "heteroarylalkyl" is a technical term and as used herein refers to an alkyl substituted with a heteroaryl, which is attached to the parent molecule moiety through the alkyl.

[0101] As used herein, the term "alkoxy" means an alkyl as defined herein attached to the parent molecule moiety through an oxygen atom. Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2 - propoxy, butoxy, tert - butoxy, pentyloxy, and hexyloxy.

[0102] The term "alkoxyalkyl" refers to an alkyl substituted with an alkoxy.

[0103] The term "alkoxycarbonyl" means an alkoxy as defined herein attached to the parent molecule moiety through a carbonyl represented by -C(=O)- as defined herein. Representative examples of alkoxycarbonyl include, but are not limited to, methoxycarbonyl, ethoxycarbonyl, and tert - butoxycarbonyl.

[0104] As used herein, the term "alkylcarbonyl" means an alkyl as defined herein attached to the parent molecule moiety through a carbonyl as defined herein. Representative examples of alkylcarbonyl include, but are not limited to, acetyl, 1 - oxopropyl, 2,2 - dimethyl - 1 - oxopropyl, 1 - oxobutyl, and 1 - oxopentyl.

[0105] As used herein, the term "arylcarbonyl" means an aryl as defined herein attached to the parent molecule moiety through a carbonyl as defined herein. Representative examples of arylcarbonyl include, but are not limited to, benzoyl and (2 - pyridyl)carbonyl.

[0106] As used herein, the terms "alkylcarbonyloxy" and "arylcarbonyloxy" mean an alkylcarbonyl or arylcarbonyl as defined herein attached to the parent molecular moiety through an oxygen atom. Representative examples of alkylcarbonyloxy include, but are not limited to, acetoxy, ethylcarbonyloxy, and tert-butylcarbonyloxy. Representative examples of arylcarbonyloxy include, but are not limited to, phenylcarbonyloxy.

[0107] The term "alkenoxy" or "alkenoxyl" means an alkenyl as defined herein attached to the parent molecular moiety through an oxygen atom. Representative examples of alkenoxy include, but are not limited to, 2-propen-1-oxy (i.e., CH2=CH-CH2-O-) and vinyloxy (i.e., CH2=CH-O-).

[0108] As used herein, the term "aryloxy" means an aryl as defined herein attached to the parent molecular moiety through an oxygen atom.

[0109] As used herein, the term "heteroaryloxy" means a heteroaryl as defined herein attached to the parent molecular moiety through an oxygen atom.

[0110] As used herein, the term "carbocyclic group" means a monocyclic or polycyclic (e.g., bicyclic, tricyclic, etc.) hydrocarbon group containing 3 to 12 carbon atoms, which is completely saturated or has one or more unsaturated bonds, and for the avoidance of doubt, the unsaturation does not give rise to an aromatic ring system (e.g., phenyl). Examples of carbocyclic groups include 1-cyclopropyl, 1-cyclobutyl, 2-cyclopentyl, 1-cyclopentenyl, 3-cyclohexyl, 1-cyclohexenyl, and 2-cyclopentenylmethyl.

[0111] The term "cyano" is a technical term and, as used herein, refers to –CN.

[0112] The term "halo" is a technical term and, as used herein, refers to –F, –Cl, -Br, or –I.

[0113] As used herein, the term "haloalkyl" means an alkyl as defined herein in which some or all of the hydrogens are replaced by halogen atoms.

[0114] The term "hydroxy" is a technical term and, as used herein, refers to –OH.

[0115] As used herein, the term "hydroxyalkyl" means that at least one hydroxy as defined herein is attached to the parent molecular moiety through an alkyl as defined herein. Representative examples of hydroxyalkyl include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, 2,3-dihydroxypentyl, and 2-ethyl-4-hydroxyheptyl.

[0116] As used herein, the term "silyl" includes hydrocarbyl derivatives of the silyl (H3Si-) group (i.e., (hydrocarbyl)3Si–), where the hydrocarbyl is a monovalent group formed by removing a hydrogen atom from a hydrocarbon, such as ethyl, phenyl. The hydrocarbyl can be a combination of different groups, and these groups can vary to provide many silyls, such as trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBS / TBDMS), triisopropylsilyl (TIPS), and [2-(trimethylsilyl)ethoxy]methyl (SEM).

[0117] As used herein, the term "silyloxy" means that a silyl as defined herein is attached to the parent molecule through an oxygen atom.

[0118] It should be understood that "substituted" or "substituted with" includes the implicit condition that such substitution conforms to the allowed valences of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., the compound does not spontaneously undergo transformation such as by rearrangement, cleavage, decomposition, cyclization, elimination, or other reactions.

[0119] The term "substituted" also contemplates all permissible substituents of organic compounds. In a broad sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Exemplary substituents include, for example, those described above herein. Permissible substituents can be one or more substituents and can be the same or different for a suitable organic compound. For the purposes of this invention, heteroatoms such as nitrogen can have hydrogen substituents and / or any permissible substituents of organic compounds that satisfy the valence of the heteroatom. The present invention is not intended to be limited in any way by the permissible substituents of organic compounds.

[0120] In certain embodiments, optional substituents can include, for example, halogen, haloalkyl, hydroxy, carbonyl (such as carboxyl (-COOH), alkoxycarbonyl, formyl or acyl), thiocarbonyl (such as thioester, thioacetate or thiocarboxylate), alkoxy, alkenyloxy, alkynyloxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (including alkylamino and dialkylamino), amido, amidine, imine, cyano, nitro, oxo (=O), azido, mercapto, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, silyl, silyloxy, heterocycloalkyl, cycloalkyl, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl or heteroarylalkyl.

[0121] For the purposes of the present invention, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Edition, 1986-87, inside front cover.

[0122] Other chemical terms used herein are used according to their conventional usage in the art, as illustrated in the McGraw-Hill Dictionary of Chemical Terms (edited by Parker, S., 1985), McGraw-Hill, San Francisco, the contents of which are incorporated herein by reference). Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0123] As used herein, the term "pharmaceutically acceptable salt" includes salts derived from inorganic or organic acids, such inorganic or organic acids including, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, phosphoric acid, formic acid, acetic acid, lactic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, glycolic acid, salicylic acid, citric acid, methanesulfonic acid, benzenesulfonic acid, benzoic acid, malonic acid, trifluoroacetic acid, trichloroacetic acid, naphthalene-2-sulfonic acid, and other acids. Pharmaceutically acceptable salt forms can include forms in which the molecular ratio of the salt present is not 1:1. For example, the salt can contain more than one inorganic or organic acid molecule per molecule of base, such as two molecules of hydrochloric acid per molecule of a compound of formula I. As another example, the salt can contain less than one inorganic or organic acid molecule per molecule of base, such as two molecules of a compound of formula I per molecule of tartaric acid.

[0124] As used herein, the terms "carrier" and "pharmaceutically acceptable carrier" refer to a diluent, adjuvant, excipient, or vehicle with which a compound is administered or formulated for administration. Non-limiting examples of such pharmaceutically acceptable carriers include liquids, such as water, saline, and oils; and solids, such as gum arabic, gelatin, starch paste, talc, keratin, colloidal silica, urea, and the like. Additionally, adjuvants, stabilizers, thickeners, lubricants, flavoring agents, and coloring agents can be used. Other examples of suitable pharmaceutical carriers are described in Remington’s Pharmaceutical Sciences by E.W. Martin, the entire contents of which are incorporated herein by reference.

[0125] As used herein, the term "treating" means preventing a disease or disorder in a subject, halting or slowing the development of the disease or disorder, or eliminating the disease or disorder. In one embodiment, "treating" means halting or slowing the development of a disease or disorder in a subject or eliminating the disease or disorder. In one embodiment, "treating" means reducing at least one objective manifestation of a disease or disorder in a subject.

[0126] As used herein, the term "effective amount" means an amount sufficient to produce the desired biological effect.

[0127] As used herein, the term "therapeutically effective amount" means an amount sufficient to produce the desired therapeutic effect.

[0128] As used herein, the term "inhibit" means to reduce an objectively measurable amount or degree. In various embodiments, "inhibit" means a reduction of at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95% compared to a relevant control. In one embodiment, "inhibit" means a 100% reduction, i.e., cessation or elimination.

[0129] As used herein, the term "subject" refers to a mammal. In various embodiments, the subject is a mouse, rat, rabbit, cat, dog, pig, sheep, horse, cow or non-human primate. In one embodiment, the subject is a human.

[0130] In the compounds of the present invention, any atom not specifically designated as a particular isotope means any stable isotope of that atom. Unless otherwise indicated, when a position is specifically designated as "H" or "hydrogen", that position is understood to have hydrogen with the natural abundance isotope composition. Additionally, unless otherwise indicated, when a position is specifically designated as "D" or "deuterium", that position is understood to have a deuterium abundance at least 3340 times greater than the natural abundance of deuterium, the natural abundance of deuterium being 0.015% (i.e., at least 50.1% deuterium incorporation).

[0131] As used herein, the term "isotope enrichment factor" means the ratio of the isotope abundance to the natural abundance of the designated isotope.

[0132] In various embodiments, the compounds of the present invention have an isotope enrichment factor of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation) or at least 6633.3 (99.5% deuterium incorporation) for each designated deuterium atom.

[0133] Pharmaceutical composition

[0134] The compositions and methods of the present invention can be used to treat an individual in need. In certain embodiments, the individual is a mammal, such as a human or a non-human mammal. When administered to an animal such as a human, the composition or compound is preferably administered in the form of a pharmaceutical composition, which comprises, for example, a compound of the present invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiological buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil or injectable organic esters. In a preferred embodiment, when such a pharmaceutical composition is administered to a human, especially for invasive administration routes (i.e., routes such as injection or implantation that circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients can be selected, for example, to achieve delayed release of the agent or to selectively target one or more cells, tissues, or organs. The pharmaceutical composition can be in dosage unit form, such as tablets, capsules (including dispersed capsules and gelatin capsules), granules, lyophilized agents for reconstitution, powders, solutions, syrups, suppositories, injections, etc. The composition can also be present in a transdermal delivery system, such as a skin patch. The composition can also be present in solutions suitable for topical application, such as lotions, creams, or ointments.

[0135] The pharmaceutically acceptable carrier can contain physiologically acceptable agents that, for example, serve to stabilize a compound (such as a compound of the present invention), increase its solubility, or increase its absorption. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose, or dextran; antioxidants such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins or other stabilizers or excipients. The choice of the pharmaceutically acceptable carrier (including the physiologically acceptable agent) depends, for example, on the route of administration of the composition. The formulation or pharmaceutical composition can be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (formulation) can also be a liposome or other polymeric matrix, in which, for example, a compound of the present invention can be incorporated. For example, liposomes containing phospholipids or other lipids are non-toxic, physiologically acceptable, and metabolizable carriers, and these carriers are relatively simple to prepare and administer.

[0136] As used herein, the phrase “pharmaceutically acceptable” refers to those compounds, materials, compositions, and / or dosage forms that are within the scope of reasonable medical judgment, suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, and commensurate with a reasonable benefit / risk ratio.

[0137] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) gum tragacanth powder; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

[0138] The pharmaceutical composition (formulation) can be administered to a subject by any of a variety of routes of administration, including, for example, oral (e.g., infusion in an aqueous or non-aqueous solution or suspension for administration to the tongue, tablets, capsules (including dispersed capsules and gelatin capsules), boluses, powders, granules, pastes); absorption through the oral mucosa (e.g., sublingual); subcutaneous; transdermal (e.g., as a patch applied to the skin); and topical administration (e.g., as a cream, ointment, or spray applied to the skin). The compound can also be formulated for inhalation. In certain embodiments, the compound can simply be dissolved or suspended in sterile water. Details of suitable routes of administration and the compositions suitable therefor can be found, for example, in U.S. Patent Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970, and 4,172,896 and the patents cited therein.

[0139] The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well-known in the art of pharmacy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host to be treated, the particular mode of administration. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, in one hundred parts, this amount will range from about 1% to about 99% active ingredient, preferably from about 5% to about 70%, most preferably from about 10% to about 30%.

[0140] Methods of preparing these formulations or compositions include the step of associating the active compound, such as a compound of the present invention, with a carrier and optionally one or more accessory ingredients. Generally, the formulations are prepared by uniformly and intimately associating the compound of the present invention with a liquid carrier, or a finely divided solid carrier, or both, and then, if necessary, shaping the product.

[0141] Formulations of the present invention suitable for oral administration may be presented in the form of capsules (including dispersed capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavoured base, usually sucrose and acacia or tragacanth), lyophilisates, powders, granules or as a solution or suspension in an aqueous liquid or a non-aqueous liquid, or as an oil-in-water or water-in-oil emulsion, or as an elixir or syrup, or as a pastille (using an inert base such as gelatin and glycerin, or sucrose and acacia) and / or as a mouthwash etc., each containing a predetermined amount of a compound of the present invention as an active ingredient. The compositions or compounds may also be administered as boluses, electuaries or pastes.

[0142] For the preparation of solid dosage forms for oral administration (capsules (including dispersed capsules and gelatin capsules), tablets, pills, dragees, powders, granules, etc.), the active ingredient is mixed with one or more pharmaceutically acceptable carriers such as sodium citrate or calcium phosphate dibasic and / or any of the following: (1) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants such as glycerol; (4) disintegrants such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarders such as paraffin wax; (6) absorption accelerators such as quaternary ammonium compositions; (7) wetting agents such as cetyl alcohol and glycerol monostearate; (8) absorbents such as kaolin and bentonite clay; (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents such as modified and unmodified cyclodextrins; and (11) colorants. In the case of capsules (including dispersed capsules and gelatin capsules), tablets, and pills, the pharmaceutical composition may also contain buffering agents. Similar types of solid compositions may also be employed as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose or lactose and high molecular weight polyethylene glycols, etc.

[0143] Tablets can be prepared by compression or molding, optionally containing one or more accessory ingredients. Compressed tablets can be prepared using binders (e.g., gelatin or hydroxypropylmethyl cellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surfactants, or dispersing agents. Molded tablets can be prepared by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0144] Solid dosage forms of tablets and other pharmaceutical compositions such as dragees, capsules (including dispersed capsules and gelatin capsules), pills, and granules may optionally be scored or prepared with coatings and shells, such as enteric coatings or other coatings well known in the pharmaceutical formulation art. They may also be formulated with, for example, different proportions of hydroxypropyl methylcellulose, other polymer matrices, liposomes, and / or microspheres for providing slow release or controlled release of the active ingredient contained therein. They may be sterilized, for example, by filtration through a bacteria-retaining filter or by incorporation of a sterilizing agent in the form of a sterile solid composition which can be dissolved in sterile water or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may have a composition which releases the active ingredient only, or preferentially, in a particular part of the gastrointestinal tract, optionally in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes. The active ingredient may also be in microencapsulated form, optionally with one or more of the above excipients where appropriate.

[0145] Liquid dosage forms suitable for oral administration include pharmaceutically acceptable emulsions, lyophilized agents for reconstitution, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, cyclodextrins and their derivatives, solubilizing agents, and emulsifying agents such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol, oils (specifically cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan and mixtures thereof.

[0146] In addition to the inert diluent, the oral compositions may also contain adjuvants such as wetting agents, emulsifying agents, and suspending agents, sweetening agents, flavoring agents, coloring agents, aromatic agents, and preservatives.

[0147] In addition to the active compound, the suspensions may also contain suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, and tragacanth and mixtures thereof.

[0148] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffers, or propellants as may be required.

[0149] In addition to the active compound, ointments, pastes, creams, and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicic acid, talc, and zinc oxide or mixtures thereof.

[0150] In addition to the active compound, the powders and sprays may also contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate and polyamide powder or mixtures of these substances. The sprays may additionally contain conventional propellants such as chlorofluorocarbons and volatile unsubstituted hydrocarbons (such as butane and propane).

[0151] Transdermal patches have the additional advantage of providing controlled delivery of the compounds of the present invention to the body. Such dosage forms may also be prepared by dissolving or dispersing the active compound in a suitable medium. Penetration enhancers may also be used to increase the flux of the compound through the skin. The rate of this flux may be controlled by providing a rate controlling membrane or by dispersing the compound in a polymeric matrix or gel.

[0152] As used herein, the phrases "parenteral administration" and "administered parenterally" mean modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions immediately before use, said combinations may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0153] Examples of suitable aqueous and non-aqueous carriers that may be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters (such as ethyl oleate). Appropriate fluidity may be maintained, for example, by using coating materials such as lecithin, by maintaining the desired particle size in the case of dispersions, and by using surfactants.

[0154] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by including various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, etc. It may also be desirable to include isotonic agents such as sugars, sodium chloride, etc. in the compositions. In addition, prolonged absorption of injectable drug forms may be achieved by including agents that delay absorption such as aluminum monostearate and gelatin.

[0155] In some cases, to prolong the action of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a poorly water-soluble crystalline or amorphous material. The absorption rate of the drug then depends on its dissolution rate, which in turn can depend on crystal size and crystal form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

[0156] Injectable depot forms are prepared by forming a microencapsulation matrix of the subject compound in a biodegradable polymer such as poly(lactide - co - glycolide). The drug release rate can be controlled according to the ratio of the drug to the polymer and the nature of the specific polymer used. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injection formulations can also be prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

[0157] For use in the methods of the present invention, the active compound can be provided per se or as a pharmaceutical composition containing, for example, from 0.1% to 99.5% (more preferably from 0.5% to 90%) of the active ingredient in combination with a pharmaceutically acceptable carrier.

[0158] The method of introduction can also be provided by a rechargeable or biodegradable device. Regarding the controlled delivery of drugs (including protein biopharmaceuticals), various sustained - release polymer devices have been developed and tested in vivo in recent years. A variety of biocompatible polymers, including hydrogels, both biodegradable and non - biodegradable, can be used to form implants for the sustained release of compounds at specific target sites.

[0159] The actual dosage level of the active ingredient in the pharmaceutical composition can be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, and that is non - toxic to the patient.

[0160] The selected dosage level depends on a variety of factors including the activity of the specific compound or combination of compounds or their esters, salts, or amides used, the route of administration, the time of administration, the excretion rate of the specific compound used, the duration of the treatment, other drugs, compounds, and / or materials used in combination with the specific compound employed, the age, sex, weight, medical condition, general health status, and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0161] A physician or veterinarian of ordinary skill in the art can readily determine and prescribe a therapeutically effective amount of the desired pharmaceutical composition. For example, the physician or veterinarian can initiate the dosage of the pharmaceutical composition or compound at a level below that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. "Therapeutically effective amount" means the concentration of a compound sufficient to elicit the desired therapeutic effect. It will generally be understood that the effective amount of a compound will vary according to the weight, sex, age, and medical history of the subject. Other factors that can affect the effective amount can include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent being co-administered with the compounds of the invention. Larger total doses can be delivered by administering the agent multiple times. Methods for determining efficacy and dosage are known to those of skill in the art (Isselbacher et al. (1996) Harrison’s Principles of Internal Medicine 13th ed., 1814-1882, incorporated herein by reference).

[0162] Generally, the suitable daily dose of the active compound used in the compositions and methods of the invention will be the amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such effective dose will generally depend on the aforementioned factors.

[0163] If desired, the effective daily dose of the active compound can be administered as one, two, three, four, five, six or more sub-doses, administered at appropriate intervals throughout the day, optionally in unit dosage forms. In certain embodiments of the invention, the active compound can be administered two or three times per day. In a preferred embodiment, the active compound will be administered once per day.

[0164] The patients to receive such treatment are any animals in need, including primates, particularly humans; and other mammals such as horses, cows, pigs, sheep, cats and dogs; poultry; and generally pets.

[0165] In certain embodiments, the compounds of the invention can be used alone or in combination with another type of therapeutic agent.

[0166] The present disclosure includes the use of pharmaceutically acceptable salts of the compounds of the present invention in the compositions and methods of the present invention. In certain embodiments, the salts contemplated by the present invention include, but are not limited to, alkyl, dialkyl, trialkyl or tetraalkyl ammonium salts. In certain embodiments, the salts contemplated by the present invention include, but are not limited to, L-arginine, benenthamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucosamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine and zinc salts. In certain embodiments, the salts contemplated by the present invention include, but are not limited to, Na, Ca, K, Mg, Zn or other metal salts. In certain embodiments, the salts contemplated by the present invention include, but are not limited to, 1-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, L-ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid / decanoic acid, caproic acid / hexanoic acid, caprylic acid / octanoic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, D-glucoheptonic acid, D-gluconic acid, D-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, L-malic acid, malonic acid, mandelic acid, methanesulfonic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, L-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid and undecylenate.

[0167] Pharmaceutically acceptable acid addition salts may also exist in the form of various solvates, such as solvates with water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates can also be prepared. The source of such solvates can be the solvent from crystallization, inherent in the solvent used for preparation or crystallization, or insoluble in such solvents.

[0168] Wetting agents, emulsifying agents and lubricants such as sodium lauryl sulfate and magnesium stearate, and coloring agents, release agents, coating agents, sweetening agents, flavoring agents and fragrances, preservatives and antioxidants may also be present in the composition.

[0169] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0170] Compounds of the present disclosure

[0171] In certain aspects, the present invention provides a compound of formula I:

[0172]

[0173] or a pharmaceutically acceptable salt thereof;

[0174] wherein:

[0175] R 1 is selected from the group consisting of: H, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl and -O-heterocycloalkyl;

[0176] R 2 is selected from the group consisting of: H, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl and -O-heterocycloalkyl; and

[0177] R 3 is selected from the group consisting of: H, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl and -O-heterocycloalkyl;

[0178] R 4 is selected from the group consisting of: H, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl and -O-heterocycloalkyl; and

[0179] R 5 is selected from the group consisting of: H, halo, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl and -O-heterocycloalkyl;

[0180] wherein when R 1 、R 2 、R3 、R 4 or R 5 is a C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl or -O-heterocycloalkyl, at least one hydrogen atom in the C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl or -O-heterocycloalkyl is replaced by a deuterium atom; and

[0181] wherein R 1 、R 2 、R 3 and R 4 at least one of them is not H.

[0182] In certain embodiments, R 1 is selected from the group consisting of: methyl, ethyl, propyl and butyl. In a further embodiment, R 1 is selected from the group consisting of: CD3, C2D5, C3D7 and C4D9. In yet a further embodiment, R 1 is: In an even further embodiment, R 1 is: In certain embodiments, R 1 is: In a further embodiment, R 1 is: In yet a further embodiment, R 1 is: In an even further embodiment, R 1 is:

[0183] In certain embodiments, R 1 is selected from the group consisting of: -OMe, -OEt, -OPr and -OBu. In a further embodiment, R 1 is selected from the group consisting of: -OCD3, -OC2D5, -OC3D7 and -OC4D9. In yet a further embodiment, R 1 is: In an even further embodiment, R 1 is:

[0184] In certain embodiments, R 2 is selected from the group consisting of: methyl, ethyl, propyl and butyl. In a further embodiment, R 2 is selected from the group consisting of: CD3, C2D5, C3D7 and C4D9. In yet a further embodiment, R 2 is: In a further embodiment, R 2 is: In certain embodiments, R 2 is: In a further embodiment, R 2 is: In yet a further embodiment, R 2 is: In a further embodiment, R 2 is:

[0185] In certain embodiments, R 2 is selected from the group consisting of: -OMe, -OEt, -OPr, and -OBu. In a further embodiment, R 2 is selected from the group consisting of: -OCD3, -OC2D5, -OC3D7, and -OC4D9. In yet a further embodiment, R 2 is: In a further embodiment, R 2 is:

[0186] In certain embodiments, R 3 is selected from the group consisting of: methyl, ethyl, propyl, and butyl. In a further embodiment, R 3 is selected from the group consisting of: CD3, C2D5, C3D7, and C4D9. In yet a further embodiment, R 3 is: In a further embodiment, R 3 is: In certain embodiments, R 3 is: In a further embodiment, R 3 is: In yet a further embodiment, R 3 is: In a further embodiment, R 3 is:

[0187] In certain embodiments, R 3 is selected from the group consisting of: -OMe, -OEt, -OPr, and -OBu. In a further embodiment, R 3 is selected from the group consisting of: -OCD3, -OC2D5, -OC3D7, and -OC4D9. In yet a further embodiment, R 3 is: In a further embodiment, R 3 is:

[0188] In certain embodiments, R 4 is selected from the group consisting of: methyl, ethyl, propyl, and butyl. In further embodiments, R 4 is selected from the group consisting of: CD3, C2D5, C3D7, and C4D9. In still further embodiments, R 4 is: In yet further embodiments, R 4 is: In certain embodiments, R 4 is: In further embodiments, R 4 is: In still further embodiments, R 4 is: In yet further embodiments, R 4 is:

[0189] In certain embodiments, R 4 is selected from the group consisting of: -OMe, -OEt, -OPr, and -OBu. In further embodiments, R 4 is selected from the group consisting of: -OCD3, -OC2D5, -OC3D7, and -OC4D9. In still further embodiments, R 4 is: In yet further embodiments, R 4 is:

[0190] In certain embodiments, R 5 is H. In additional embodiments, R 5 is a halogen group. In still further embodiments, R 5 is F. In yet further embodiments, R 5 is selected from the group consisting of: methyl, ethyl, propyl, and butyl. In certain embodiments, R 5 is selected from the group consisting of: CD3, C2D5, C3D7, and C4D9. In further embodiments, R 5 is: In still further embodiments, R 5 is: In yet further embodiments, R 5 is: In certain embodiments, R 5 is:

[0191] In a further embodiment, R 5 is: In yet a further embodiment, R 5 is:

[0192]

[0193] In certain embodiments, R 5 is selected from the group consisting of: -OMe, -OEt, -OPr, and -OBu. In a further embodiment, R 5 is selected from the group consisting of: -OCD3, -OC2D5, -OC3D7, and -OC4D9. In yet a further embodiment, R 5 is: In an even further embodiment, R 5 is:

[0194] In certain embodiments, the compound is selected from the group consisting of:

[0195]

[0196] or a pharmaceutically acceptable salt thereof.

[0197] In a further embodiment, the compound is selected from the group consisting of:

[0198]

[0199] or a pharmaceutically acceptable salt thereof.

[0200] In certain embodiments, the compound is selected from the group consisting of:

[0201]

[0202] or a pharmaceutically acceptable salt thereof.

[0203] In a further aspect, the present disclosure provides a pharmaceutical composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient. In certain embodiments, the pharmaceutical composition further comprises an additional therapeutic agent selected from an iron chelator and deferiprone.

[0204] Treatment methods

[0205] In yet a further aspect, the present disclosure provides a method of treating a disease or disorder characterized by reduced ferroportin, comprising administering to a subject in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure. In certain embodiments, the disease or disorder characterized by reduced ferroportin is an age-related, induced ferroportin disease.

[0206] In a further aspect, the present disclosure provides methods of treating a disease or disorder characterized by reduced ferroportin, which comprise administering to a subject in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure.

[0207] In certain aspects, the present disclosure provides methods of treating a disease or disorder characterized by a ferroportin defect or deficiency, which comprise administering to a subject in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure.

[0208] In a further aspect, the present disclosure provides methods of treating a disease or disorder characterized by iron accumulation, which comprise administering to a subject in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure. In certain embodiments, the iron accumulation is iron accumulation in the brain.

[0209] In yet a further aspect, the present disclosure provides methods of treating a disease or disorder characterized by improper iron distribution, which comprise administering to a subject in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure. In certain embodiments, the improper iron distribution is improper iron distribution in the brain.

[0210] In certain embodiments, the disease or disorder is selected from the group consisting of: inflammatory anemia, iron overload, thalassemia, hemochromatosis, atransferrinemia, myelodysplasia, hypochromic microcytic anemia, ferroportin disease, transfusional iron overload, and aceruloplasminemia.

[0211] In some aspects, the present disclosure provides methods for treating neurodegenerative diseases, which include administering to a subject in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure. In certain embodiments, the neurodegenerative disorder is selected from the group consisting of Parkinson’s Disease, Alzheimer’s Disease, amyotrophic lateral sclerosis, Friedrich’s ataxia, Huntington’s Disease, Lewy Body Disease, and spinal muscular atrophy. In a further embodiment, the neurodegenerative disorder is neurodegeneration with brain iron accumulation (NBIA). In a still further embodiment, NBIA is selected from the group consisting of beta-propeller protein-associated neurodegeneration (BPAN), pantothenate kinase-associated neurodegeneration (PKAN), PLA2G6-associated neurodegeneration (PLAN), mitochondrial membrane protein-associated neurodegeneration (MPAN), fatty acid hydroxylase-associated neurodegeneration (FAHN), COASY protein-associated neurodegeneration (CoPAN), aceruloplasminemia, Kufor-Rakeb syndrome (also known as Parkinson disease 9 (PARK9)), neuroferritinopathy, Woodhouse-Sakati syndrome, and idiopathic NBIA.

[0212] In certain embodiments, the methods of the present disclosure further include administering to the subject an effective amount of an additional therapeutic agent selected from iron chelators and deferiprone.

[0213] Examples

[0214] The invention has been generally described above. The invention will be more readily understood by reference to the following examples, which are included solely for the purpose of illustrating certain aspects and embodiments of the invention and are not intended to limit the invention.

[0215] Example 1 - Preparation of boronic acid derivatives

[0216] Add magnesium turnings (1 equiv) to a dried 40 mL vial equipped with a stir bar, and then evacuate the vial three times with N2. Under N2, add the deuterated alkyl halide and dry Et2O to a second dried 40 mL vial equipped with a stir bar. Briefly mix the second vial before adding the mixture dropwise to the Mg-containing vial. While allowing the Grignard reagent to form over 2 hours, place a third dried vial equipped with a stir bar under N2 and add trimethyl borate (1.2 equiv) and dry Et2O. Then cool the third vial to -78 °C and add the Grignard reagent dropwise over a 10-minute period. Stir the solution at -78 °C for 20 minutes and then warm to 23 °C over 1 hour. Add 1 M HCl solution and stir the solution at 23 °C for 30 minutes. Subsequently, extract the reaction mixture with Et2O (2 × 6 mL), wash with brine (10 mL), and dry over Na2SO4. Add 0.5 mL of dry dioxane to the dried solution and remove the ether under a stream of N2. The dioxane containing the crude boronic acid is used directly in the Suzuki-Miyuara coupling with the bromotropolone conjugate.

[0217] Example 2 - Synthesis of bromocycloheptatrienone scaffolds

[0218] α-bromotropolone scaffold

[0219]

[0220] A dried two-neck round-bottom flask equipped with a condenser is evacuated and backfilled three times with N2. Add anhydrous K2CO3 (3 equiv), tropolone (1 equiv), and 18-crown-6 (0.1 equiv), followed by MeCN. Then add MeI (5 equiv) and reflux the reaction mixture for 5 hours. Cool the reaction mixture and then filter through a Celite pad, rinsing with MeCN. Then concentrate the filtrate in vacuo and purify by flash column chromatography to afford methyl-protected α-bromotropolone 1a.

[0221] 1 H NMR (500 MHz, CDCl3) δ 7.28–7.18 (m, 2H), 7.10 (ddd, J = 10.8, 9.8, 1.0 Hz, 1H), 6.91–6.84 (m, 1H), 6.77 (dd, J = 9.9, 0.8 Hz, 1H), 3.94 (s, 3H).

[0222] 13 C NMR (126 MHz, CDCl3) 180.2, 165.1, 136.5, 136.5, 132.7, 127.8, 112.3, 56.1

[0223] Calculated HRMS(ESI+) for C8H8O2[M+H]: 137.0603, Observed: 137.0603

[0224]

[0225] The previous product (1 equiv) was added to a 40 mL chemical vial equipped with a stir bar, followed by the addition of NBS (1.2 equiv) (recrystallized from water) and CCl4. The reactants were then heated at 90 °C for 3 h. The reactants were then rinsed into a separatory funnel with DCM, washed with saturated sodium thiosulfate, and then extracted 3 times with DCM. The combined organic extracts were washed with brine, dried over MgSO4, and then concentrated in vacuo. The product was then purified by flash chromatography to afford the α-bromotropone scaffold 1b.

[0226] 1 H NMR (500 MHz, CDCl3) δ 8.19 (dd, J = 9.5, 1.0 Hz, 1H), 7.15 (ddd, J = 10.8, 9.9, 1.0 Hz, 1H), 6.79 (dt, J = 9.9, 0.7 Hz, 1H), 6.68 (ddd, J = 10.4, 9.5, 0.7 Hz, 1H), 3.96 (d, J =.8 Hz, 3H)

[0227] 13 C NMR (126 MHz, CDCl3) δ 174.0, 162.9, 140.0, 138.0, 133.0, 125.4, 112.4, 56.9

[0228] Calculated HRMS(ESI+) for C8H7O2Br[M+H]: 214.9708, Observed: 214.9717

[0229] β-bromotropone scaffold

[0230]

[0231] A large round-bottom flask was equipped with an addition funnel, evacuated, and backfilled with N2 three times. KOtBu (1.5 equiv) and pentane were added to the flask. The solution was then immersed in an ice bath and cooled to 0 °C. Then, 1,3-cyclohexadiene (1 equiv) was added via syringe. Then, CHBr3 (1.2 equiv) was added via the addition funnel over the course of 1 h. The reactants were then stirred at 0 °C for 1 h, then warmed to room temperature and stirred for an additional 1 h. Then, water was added to the round-bottom and stirred, and the reactants were transferred to a separatory funnel, extracted 3 times with hexane, dried over MgSO4, and concentrated in vacuo. The product was then purified by flash chromatography to afford the desired intermediate 2a as an oil.

[0232] 1 HNMR(500MHz, CDCl3) δ 5.92 (app. d, J = 2.9 Hz, 2H), 2.14 (dd, J = 10.4, 2.2 Hz, 1H), 2.08 - 2.01 (m, 3H), 1.95 - 1.90 (m, 2H)

[0233] 13 C NMR(126MHz, CDCl3) δ 130.1, 122.2, 39.6, 29.6, 27.7, 21.0, 17.8

[0234] HRMS(ESI+) calcd for C7H8Br2[M + H]: 249.8993, found: 249.8996

[0235]

[0236] To a 40 mL chemical vial was added 2a (1 equiv), followed by the addition of a solution containing 1:1 tBuOH:H2O, NMO (2 equiv) and freshly distilled pyridine (2.5 equiv). Then, OsO4 (0.03 equiv) was added as a 40 mg / mL solution from H2O. The reaction mixture was then heated to 100 °C and stirred vigorously for 3 h. The reaction mixture was then cooled and poured into an aqueous sodium thiosulfate solution and quenched overnight. The crude product was then added to a separatory funnel and extracted 5 times with Et2O. The organic extract was washed with saturated aqueous CuSO4, water and brine, then dried over MgSO4 and concentrated in vacuo. The crude product was then purified by flash column chromatography to afford the desired product 2b as a brown solid.

[0237] 1 H NMR(500MHz, CD3OD) δ 3.77 (ddd, J = 5.6, 3.0, 1.9 Hz, 1H), 3.5 (t, J = 2.8 Hz, 1H), 2.12 - 2.03 (m, 1H), 2.03 - 1.96 (m, 1H), 1.85 (dd, J = 11.1, 2.6 Hz, 1H), 1.70 - 1.54 (m, 2H), 1.41 - 1.26 (m, 1H).

[0238] 13 C NMR(126MHz, CD3OD) δ 69.5, 67.8, 37.2, 34.2, 29.1, 26.6, 16.7

[0239] C7H 10 HRMS(ESI+) calcd for C7H8O2Br2[M + Na]: 306.8945, found: 306.8948

[0240]

[0241] The dried three-necked round-bottom flask was equipped with a stir bar and placed under N2. DCM (200 mL) and DMSO (4 equiv) were added, and the reactants were cooled to -65 °C using an immersion cooler. After cooling, TFAA (4 equiv) was added via syringe over 10 minutes. After 15 minutes, a solution of 2b (1 equiv) was added to DMSO (10 mL) over 10 minutes. The reaction was allowed to proceed for 2 hours, then NEt3 (7 equiv) was added over 10 minutes. The reaction was allowed to proceed for 2 hours, then warmed to room temperature. The reaction was then quenched with 200 mL of 2 M HCl, transferred to a separatory funnel and extracted 3 times with DCM. The combined organic layers were then washed three times with water, then once with brine, then dried over MgSO4 and concentrated in vacuo. This mixture containing 2c was then taken to the next step without purification.

[0242]

[0243] The dried two-necked round-bottom flask was equipped with a condenser, evacuated and backfilled with N2 three times. Anhydrous K2CO3 (3 equiv), the mixture of 2c (1 equiv) and 18-crown-6 (0.1 equiv) were added, followed by MeCN. Then MeI (5 equiv) was added, and the reactants were refluxed for 5 hours. The reactants were cooled, then filtered through a pad of Celite and rinsed with MeCN. The filtrate was then concentrated in vacuo and purified by flash column chromatography to give the methyl-protected β-bromotropone.

[0244] 1 H NMR (500 MHz, CDCl3) δ 7.63 (d, J = 2.0 Hz, 1H), 7.06 (dd, J = 11.4, 2.0 Hz, 1H), 6.79 (dd, J = 11.4, 10.1 Hz, 1H), 6.59 (d, J = 10.1 Hz, 1H), 3.90 (s, 3H)

[0245] 13 C NMR (126 MHz, CDCl3) δ 177.2, 165.3, 139.3, 136.9, 132.5, 131.6, 111.0, 56.5

[0246] HRMS (ESI+) calcd for C7H5O2Br [M+H]: 219.9708, found 219.9704

[0247] γ-bromotropone scaffold

[0248]

[0249] The γ-bromotropone scaffold was synthesized following the same procedure as the β-bromotropone scaffold, but using 1,4-cyclohexadiene instead of 1,3-cyclohexadiene in the first step and producing the following intermediates.

[0250]

[0251] 1 HNMR(500MHz,CDCl3)d5.50(s,2H),2.47(d,J=17.5Hz,2H),2.10(d,J=17.5Hz,2H),1.91(dd,J=5.3,2.4Hz,2H)

[0252] 13 C NMR(126MHz,CDCl3)d 122.6,38.5,25.1,21.1

[0253] HRMS(ESI+) calculated for C7H8Br2[M+H]: 249.8993, observed: 249.9005

[0254]

[0255] 1 HNMR(500MHz,CD3OD)d3.66-3.59(m,2H),2.17(dtd,J=15.3,6.6,3.1Hz,1H),1.99-1.90(m,2H),1.76(ddd,J=15.2,4.7,1.8Hz,2H)

[0256] 13 C NMR(126MHz,CD3OD)d 68.3,40.2,28.4,28.0

[0257] C7H 10 HRMS(ESI+) calculated for C7H8O2Br2[M+H]: 265.8942, observed: 265.8941

[0258]

[0259] It was used in the next step without the above purification.

[0260]

[0261] 11H NMR (500 MHz, CDCl3) δ 7.38 (dd, J = 13.1, 2.0 Hz, 1H), 7.36 (dd, J = 11.0, 2.0 Hz, 1H), 6.95 (d, J = 13.0 Hz, 1H), 6.46 (d, J = 10.9 Hz, 1H), 3.88 (s, 3H)

[0262] 13 13C NMR (126 MHz, CDCl3) δ 179.5, 164.8, 140.1, 136.1, 134.4, 122.6, 111.6, 56.5

[0263] HRMS (ESI+) calcd for C7H5O2Br2[M+H]+: 214.9708, found: 214.9710

[0264] 1,3 - dibromotropone scaffold

[0265]

[0266] Synthesis was the same as the procedure listed in 1A

[0267]

[0268] To a dried vial equipped with a stir bar was added 30 mL of CCl4 and the methyl - protected tropone (1 g, 7.3 mmol) from the previous step. N - Bromosuccinimide (2.1 equiv) was added in two portions, then the vial was capped, sealed with electrical tape and heated at 90 °C overnight. The reaction mixture was then rinsed into a separatory funnel with DCM, washed with saturated sodium thiosulfate, and then extracted with DCM three times. The combined organic extracts were washed with brine, dried over MgSO4, and then concentrated in vacuo. The product was then purified by flash chromatography to afford the desired product

[0269] 1 1H NMR (500 MHz, CDCl3) δ 8.45 (d, 1H), 7.46 (d, 1H), 6.54 (d, 1H), 3.94 (s, 3H)

[0270] 13 13C NMR (126 MHz, CDCl3) δ 173.3, 161.9, 142.8, 137.0, 134.5, 119.5, 111.5, 56.9

[0271] 1,3,5 - tribromotropone scaffold

[0272]

[0273] To a dried 40 mL vial equipped with a stir bar, add 30 mL of CCl4 and tropolone (1 g, 8.1 mmol). Add N-bromosuccinimide (3.2 equiv) in three portions, then cap the vial, seal it with electrical tape and heat to 90 °C overnight. Then rinse the reaction mixture into a separatory funnel with DCM, wash with saturated sodium thiosulfate, and then extract three times with DCM. Then wash the combined organic extracts with brine, dry over MgSO4, and then concentrate in vacuo to afford the pure product. The pure product was used directly in the next step.

[0274] 1 HNMR (500 MHz, CDCl3) δ 8.34 (s, 2H)

[0275] 13 C NMR (126 MHz, CDCl3) δ 164.4, 144.0, 124.3, 118.0

[0276]

[0277] A dried two-neck round bottom flask was equipped with a condenser, evacuated and backfilled with N2 three times. Add anhydrous potassium carbonate K2CO3 (16.97 g, 123 mmol, 3 equiv), 7a (5 g, 41 mmol, 1 equiv) and 18-crown-6 (1.08 g, 4.1 mmol, 0.1 equiv), followed by addition of MeCN (250 mL). Then add MeI (29.1 g, 205 mmol, 5 equiv), and reflux the reaction mixture for 3 h. Cool the reaction mixture, then filter through a pad of Celite and rinse with acetone. Then concentrate the filtrate in vacuo and purify by flash column chromatography to afford the product as a yellow solid.

[0278] 1 HNMR (500 MHz, CDCl3) δ 8.28 (s, 1H), 7.95 (s, 1H), 4.01 (s, 3H)

[0279] Example 3 - General procedure for derivatives containing α'-fluoro / α-substituted

[0280]

[0281] Prepare the benzyl-protected α-bromotropolone scaffold (4) as described previously. Add dry CsF (1.5 equiv) to a mixture of this scaffold in DMSO under an atmosphere of N2. Stir the mixture at 110 °C under N2 for 8 h. Cool the reaction mixture to 25 °C and add brine. Extract the aqueous mixture with EtOAc, wash the combined organic layers with brine, and then dry over Na2SO4. Concentrate the solution and purify by silica gel chromatography to afford the product.

[0282] Then, as described above, the obtained residue was deprotected using TFA at 50 °C to give the α-fluoro tropolone scaffold. Briefly, the residue was dissolved in TFA and the mixture was stirred at 50 °C for 1 h. The crude product after vacuum concentration of the mixture was used for the next step.

[0283] Under N2 at 25 °C, NBS (1.3 equiv) was slowly added to a CCl4 solution of the crude product. The mixture was heated to 80 °C and maintained for 3 h. Thereafter, the mixture was cooled to 25 °C and water was added. The aqueous mixture was extracted with 3 x dichloromethane, and the combined organic matter was washed with brine and dried over Na2SO4. After filtration, the solution was concentrated to dryness to give the fluoro / bromo intermediate as a crude mixture, which was used for the next step without purification.

[0284] Then the intermediate was reprotected using the methylation protection procedure as described above (4). Briefly, the crude residue was resuspended in MeCN, and K2CO3 (3 equiv) and 18-crown-6 (0.1 equiv) were added thereto under N2. Then MeI (4 equiv) was added dropwise and the reactants were heated to 85 °C and allowed to run for 12 h. After cooling, the mixture was filtered and the residue was concentrated under reduced pressure to give a crude product. The crude product was purified by silica gel chromatography to give the desired methylated fluoro / bromo tropolone scaffold. Then this scaffold was used for GP1 and subsequently for GP2 to give the desired α-fluoro / α-substituted derivative.

[0285] Example 4 - General procedure for derivatives containing α'-fluoro / α,γ-substituted

[0286]

[0287] After GP1, the methyl-protected α-fluoro / α-substituted derivative was obtained as described above. Under N2 at 25 °C, NBS (1.3 equiv) was slowly added to a solution of this product in CHCl3. The mixture was heated to 80 °C and maintained for 3 h. Thereafter, the mixture was cooled to 25 °C and water was added. The aqueous mixture was extracted with 3 x dichloromethane, and the combined organic matter was washed with brine and dried over Na2SO4. After filtration, the solution was concentrated to dryness to give the α-fluoro / α-substituted / γ-bromo intermediate, which was purified by silica gel chromatography. The purified product was used for GP1 and subsequently for GP2 to give the α-fluoro / α,γ-substituted derivative.

[0288] Example 5 - General procedure for derivatives containing α'-fluoro / γ-substituted

[0289]

[0290] Synthesize the γ-bromotropone scaffold as described in Example 2. Then use this scaffold in GP1 to obtain the protected coupling product. Subsequently, redissolve the product in CHCl3 and slowly add NBS (1.3 equiv) at 25 °C under N2. Heat the mixture to 80 °C for 3 hours. After that, cool the mixture to 25 °C and add water. Extract the aqueous mixture with 3 x dichloromethane, and wash the combined organic matter with brine and dry over Na2SO4. After filtration, concentrate the solution to dryness to obtain the α’-bromo / γ-substituted intermediate, which is purified by silica gel chromatography. As described previously, use the purified product for the α’-fluorination procedure.

[0291] Briefly, add dry CsF (1.5 equiv) to a mixture of the scaffold in DMSO under an atmosphere of N2. Stir the mixture at 110 °C for 8 hours under N2. Cool the reactant to 25 °C and add brine. Extract the aqueous mixture with EtOAc, wash the combined organic layers with brine, and then dry over Na2SO4. Concentrate the solution and purify it by silica gel chromatography to obtain the protected product. Then deprotect the product using GP2 to obtain the desired final product.

[0292] Example 6 - Procedure for α-perdeuterated furan derivatives

[0293]

[0294] Prepare the benzyl-protected α-bromotropone scaffold for Heck coupling as described in Example 2.

[0295] Preparation of 2’3’-dideoxyfuran-d6 (a) (based on the synthesis from J. Org. Chem. 2007, 72, 19, 7253 - 7259)

[0296]

[0297] Suspend LiAlD4 in THF and reflux for 30 minutes. After cooling to -55 °C using an immersion cooler, add a solution of succinic anhydride-d4 in THF dropwise over 30 minutes. Warm the solution to 25 °C over 90 minutes, then cool to -15 °C over 15 minutes. Slowly add 6M HCl solution, and heat the solution to 25 °C and stir for 20 minutes. Wash the reaction mixture with brine and extract the organic matter with 3 x Et2O. Dry the combined organic matter over Na2SO4 and concentrate in vacuo. Purify the residue by fractional distillation (BP 65 °C) to obtain the desired product.

[0298] The product was then dissolved in dichloromethane and cooled to -78 °C using an immersion cooler and held for 2 h. A cold solution of DIBAL-D in DCM (4 °C) was added slowly and the reaction was allowed to proceed for 2 h. The reaction was quenched by the slow addition of MeOD and saturated Rochelle salt and the reactants were warmed to 25 °C where a precipitate formed. The solid was resuspended in dichloromethane and the filtrate was concentrated to give the crude lactol which was used directly in the next step without purification.

[0299] A solution of TsOH in quinoline was charged to a three-necked flask and a distillation apparatus was assembled, with a 2 M aqueous solution of NaOH kept frozen around the bottom of the receiving flask. The quinoline solution was immersed in a wax bath at 190 °C and the receiving flask was kept at -78 °C in a dry ice / acetone bath. The crude product from the previous step was added dropwise to the quinoline solution and the solution was heated to 210 °C for 1 h. The recovered liquid was quickly removed from the apparatus and purified by distillation (BP 54 °C) to give the final desired product.

[0300] As described in previous Example 2, 2’3’-dideoxyfuran-d6 (a) and a benzyl-protected α-bromocycloheptatrienone scaffold were used in the Heck coupling.

[0301] Briefly, under Ar in a glove box, K2CO3 (2 equiv), Ph3P (0.20 equiv) and Pd(OAc)2 (0.10 equiv) were added to a mixture of a (5 equiv) and the benzyl-protected α-bromocycloheptatrienone scaffold in dioxane. The mixture was then heated to 110 °C for 2 h, after which the reactants were cooled and water was added. The mixture was extracted with 2 x EtOAc, the combined organics were washed with brine and dried over Na2SO4. The crude product was concentrated in vacuo and purified by silica gel chromatography to give the racemic Heck coupling intermediate (b).

[0302] Under N2, Rh(PPh3)3Cl was added to a solution of intermediate (b) in MeOD. The reaction flask was evacuated and refilled with D2 3x, and the mixture was stirred under D2 (1 atm) for 4 h. The reaction mixture was filtered through Celite, rinsed with MeOH, and then concentrated in vacuo to give the desired product as a racemic mixture. The mixture was resolved using chiral SFC with DAICEL CHIRALPAK AD (250 mm * 30 mm, 10 mm) as the stationary phase. The separated enantiomers were then used separately in the final step to obtain the desired product.

[0303] The enantiomers were separately dissolved in pure TFA and heated to 50 °C with stirring for 2 h. The reaction mixture was cooled and then concentrated in vacuo to give the crude product. The residue was purified separately by preparative HPLC using Nano-microKromasil C18 (100 mm * 30 mm, 8 mm) as the stationary phase to give the final desired product as the pure enantiomers.

[0304] Example 7 - General procedure for derivative preparation by Csp2-Csp3 bond formation

[0305] Generally, the fully deuterated derivatives were synthesized as follows: The bromocycloheptatrienone scaffold was synthesized as described above. A dried 8 mL vial equipped with a stir bar and a vial containing the crude boric acid were placed in an argon-filled glove box. In the glove box, all solid components were first added: bromocycloheptatrienone scaffold (0.5 mmol, 1 equiv), Pd2(dba)3 (0.05 mmol, 0.1 equiv), P(o-tol)3 (0.1 mmol, 0.2 equiv), Ag2O (0.75 mmol, 1.5 equiv). Then, an additional 0.5 mL of dry dioxane was added to the crude boric acid mixture (total 1 mL of dioxane) before adding the solution of boric acid in dioxane to the chemical vial using a micropipette. The polybrominated scaffolds utilized the reagent equivalents per bromide such that, for example, for the tribromocycloheptatrienone scaffold, Pd2(dba)3 (0.15 mmol, 0.3 equiv), P(o-tol)3 (0.3 mmol, 0.6 equiv), Ag2O (2.25 mmol, 4.5 equiv) were used in a total of 3 mL of dry dioxane. For the dibromocycloheptatrienone scaffold, Pd2(dba)3 (0.1 mmol, 0.2 equiv), P(o-tol)3 (0.2 mmol, 0.4 equiv), Ag2O (1.5 mmol, 3 equiv) were used in a total of 2 mL of dry dioxane. The vial was then capped and removed from the glove box and stirred at 85 °C for 36 h. Thereafter, the crude reaction mixture was filtered through a silica plug, rinsed with EtOAc, and then concentrated in vacuo. The mixture was then dissolved in HPLC-grade MeOH (5 mL) and then purified by preparative HPLC on an Agilent 1200 series HPLC equipped with a Waters Sunfire OBD preparative column ( 5 μm, 30 mm × 150 mm) using MeCN and H2O as the mobile phase at a flow rate of 40 mL / min and monitored at 238 nm. The purified material was collected and then concentrated in vacuo, and the remaining water was removed azeotropically with additional MeCN as needed.

[0306] Another method for forming a C-C bond between sp2 and sp3 centers involves using a Grignard reagent to start with a haloalkane counterpart to produce a Kumada conjugate (J. Organomet. Chem. 1997, 532, 1, 271 - 273; Bioorg. Med. Chem. 2007, 15, 22, 7144 - 7165; Adv. Synth. Catal. 2019, 361, 19, 4468 - 4473). 2-Hydroxy-4-(isopropyl-d7)-cyclohepta-2,4,6-trien-1-one and 2-hydroxy-5-(isopropyl-d7)-cyclohepta-2,4,6-trien-1-one can be prepared in a similar manner using fully deuterated haloalkanes.

[0307] Acid deprotection of the methyl ether coupling product.

[0308] An 8 mL chemical vial was equipped with a stir bar. The purified material from GP1 was dissolved in dioxane (0.11 M) and added to the vial. Next, 20% H2SO4 solution (0.11 M, equal volume to dioxane) was added. Then it was heated to 100 °C and stirred for 3 hours. Thereafter, the reaction mixture was diluted with water and extracted 3 times with ether. Then the combined organic extracts were dried over Na2SO4 and concentrated in vacuo to give the final product.

[0309] Example 8 - Characterization data of selected compounds

[0310]

[0311] 2-Hydroxy-3-methylcyclohepta-2,4,6-trien-1-one

[0312] 1 1H NMR (500 MHz, CDCl3) δ 7.55–7.51 (m, 1H), 7.35 (dd, J = 10.3, 1.2 Hz, 1H), 7.32–7.27 (m, 1H), 6.97 (td, J = 9.8, 1.2 Hz, 1H), 2.48 (s, 3H).

[0313] 13 13C NMR (126 MHz, CDCl3) δ 172.5, 167.6, 139.9, 138.1, 135.8, 127.4, 121.2, 22.2.

[0314] HRMS (ESI+) calculated for C8H8O2[M + H]: 137.0603, found: 137.0602

[0315]

[0316] 2-Hydroxy-4-methylcyclohepta-2,4,6-trien-1-one

[0317] 1 HNMR(500MHz,CDCl3)δ7.30-7.24(m,3H),6.92(m,1H),2.47(s,3H)

[0318] 13 C NMR(126MHz,CDCl3)δ170.8,170.7,149.9,136.7,129.7,125.7,121.9,27.2

[0319] HRMS(ESI+) calculated for C8H8O2[M+H]: 137.0603, found: 137.0607

[0320]

[0321] 2-Hydroxy-5-methylcyclohepta-2,4,6-trien-1-one

[0322] 1 HNMR(500MHz,CDCl3)δ7.32–7.26(m,4H),2.45(s,3H)

[0323] 13 C NMR(126MHz,CDCl3)δ170.7,138.4,123.9,25.9

[0324] HRMS(ESI+) calculated for C8H8O2[M+H]: 137.0603, found: 137.0600

[0325]

[0326] 2-Hydroxy-3-(methyl-d3)cyclohepta-2,4,6-trien-1-one

[0327] 1 HNMR(500MHz,CDCl3)δ7.52(d,1H),7.35(d,1H),7.29(t,1H),6.97(t,1H)

[0328] 2 H NMR(500MHz,CDCl3)δ2.45

[0329] 13 C NMR(126MHz,CDCl3)δ172.5,167.5,140.5,137.8,135.7,127.3,121.2

[0330] HRMS(ESI+) calculated value for C8H5D3O2[M+H]: 140.0791, observed value: 140.0793

[0331]

[0332] 2-Hydroxy-4-(methyl-d3)cyclohepta-2,4,6-trien-1-one

[0333] 1 HNMR(500MHz,CDCl3)δ7.30 - 7.24(m,3H),6.92(m,1H)

[0334] 2 HNMR(500MHz,CDCl3)δ2.46

[0335] 13 C NMR(126MHz,CDCl3)δ170.8,170.7,149.9,136.8,129.8,125.7,122.0

[0336] HRMS(ESI+) calculated value for C8H5D3O2[M+H]: 140.0791, observed value: 140.0796

[0337]

[0338] 2-Hydroxy-5-(methyl-d3)cyclohepta-2,4,6-trien-1-one

[0339] 1 HNMR(500MHz,CDCl3)δ7.32–7.26(m,4H)

[0340] 2 HNMR(500MHz,CDCl3)δ2.39

[0341] 13 C NMR(126MHz,CDCl3)δ170.7,138.5,124.0

[0342] HRMS(ESI+) calculated value for C8H5D3O2[M+H]: 140.0791, observed value: 140.0794

[0343]

[0344] 2-Hydroxy-1,3,5-trimethylcyclohepta-2,4,6-trien-1-one

[0345] 1HNMR(500MHz,CD3CN)δ7.39(s,2H),2.38(s,3H),2.37(s,6H)

[0346] 13 C NMR(126MHz,CD3CN)δ167.8,140.1,136.8,135.0,26.2,21.5

[0347]

[0348] 2-Hydroxy-4-(isopropyl-d7)-cyclohepta-2,4,6-trien-1-one

[0349] 1 HNMR(500MHz,CDCl3)δ7.30 - 7.24(m,3H),6.95(m,1H)

[0350] 2 HNMR(500MHz,CDCl3)δ2.90,1.27

[0351] 13 C NMR(126MHz,CDCl3)δ170.8,170.7,149.9,136.5,129.7,125.6,121.5,

[0352]

[0353] 2-Hydroxy-5-(isopropyl-d7)-cyclohepta-2,4,6-trien-1-one

[0354] 1 H NMR(500MHz,CDCl3)δ7.32–7.26(m,4H)

[0355] 2 H NMR(500MHz,CDCl3)δ2.92,1.26

[0356] 13 C NMR(126MHz,CDCl3)δ170.7,144.0,138.2,124.1

[0357]

[0358] 2-Hydroxy-4-butylcyclohepta-2,4,6-trien-1-one

[0359] 1HNMR(500MHz,CDCl3)δ7.30 - 7.24(m,3H),6.92(dd,1H),2.63(dd,2H),1.64(m,2H),1.38(tq,1H),0.94(t,1H)

[0360] 13 C NMR(126MHz,CDCl3)δ170.9,170.5,154.6,136.9,129.4,125.2,122.1,41.0,33.4,22.2,13.8

[0361]

[0362] 2-Hydroxy-4-(butyl-d9)cyclohepta-2,4,6-trien-1-one

[0363] 1 HNMR(500MHz,CDCl3)δ7.30 - 7.24(m,3H),6.92(m,1H)

[0364] 2 HNMR(500MHz,CDCl3)δ2.57,1.56,1.30,0.87

[0365] 13 C NMR(126MHz,CDCl3)δ170.9,170.7,154.6,136.9,129.4,125.2,122.1

[0366]

[0367] 2-Hydroxy-1,3-dimethylcyclohepta-2,4,6-trien-1-one

[0368] 1 HNMR(500MHz,CDCl3)δ7.43(s,1H),7.26(d,1H),7.16(d,1H),2.40(s,3H),2.37(s,3H)

[0369] 13 C NMR(126MHz,CDCl3)δ171.9,166.4,141.3,137.7,137.5,136.0,121.4,26.0,22.3

[0370]

[0371] 2-Hydroxy-1,3-(dimethyl-d6)cyclohepta-2,4,6-trien-1-one

[0372] 1 HNMR (500 MHz, CDCl3) δ 7.43 (s, 1H), 7.26 (d, 1H), 7.16 (d, 1H)

[0373] 2 HNMR (500 MHz, CDCl3) δ 2.41, 2.37

[0374] 13 C NMR (126 MHz, CDCl3) δ 171.9, 166.2, 141.4, 137.7, 137.5, 136.0, 121.3

[0375] Example 9 - Preparation of liposomes

[0376] Liposomes were prepared as described above (A.S. Grillo et al., Science, 356, 608 - 615 (2017)). A lipid film (Avanti Polar Lipids 850457C) was generated by adding 200 μL of chloroform containing 4 mg / mL recrystallized cholesterol to 960 μL of a 25 mg / mL solution of POPC in chloroform. After evaporation under a nitrogen stream, the film was placed under high vacuum for 18 h to ensure quantitative removal of the solvent. The lipid film was rehydrated by adding 1 mL of liposomes to a buffer (15 mM FeCl3, 125 mM citrate, 50 mM MES / TRIS pH 7.0) and vortexed thoroughly to form a solution of multilamellar vesicles (MLV). The liposomes were then extruded using a.2 mm membrane (Whatman 800281) through an Avanti Lipids extruder (Avanti Polar Lipids 610023) 21 times to ensure homogeneous large unilamellar vesicles (LUV). Size exclusion chromatography of the LUV was performed using a Sephadex G - 50 Medium resin column hydrated with an external buffer of the liposomes (600 mM sodium ascorbate, 50 mM MES / TRIS pH 7.0). Separation of the LUV from the extracellular Fe could be easily observed on the column, and the most concentrated fraction of the LUV was collected. Size exclusion chromatography of the LUV was performed using a Sephadex G - 50 Medium resin column hydrated with an external buffer of the liposomes (600 mM sodium ascorbate, 50 mM MES / TRIS pH 7.0). Separation of the LUV from the extracellular Fe could be easily observed on the column, and the most concentrated fraction of the LUV was collected.

[0377] Example 10 - Quantification of liposomes

[0378] The total phosphorus was determined using the Bartlett assay. Briefly, the LUV suspension was diluted 10-fold in the external liposome buffer, and then 10 μL of this solution was placed in three 7 mL vials together with 10 μL of the external liposome buffer as a background. Then 450 μL of 8.9 M H2SO4 was added, and the mixture was heated in air at 225 °C for 25 minutes. After cooling, 150 μL of 30% H2O2 was added, and the mixture was further heated in air at 225 °C for 30 minutes. Then the sample was allowed to cool completely and diluted with 3.9 mL of Milli-Q water. Then 500 μL of 2.5% w / v ammonium molybdate was added, then the sample was vortexed, and then 500 μL of 10% w / v ascorbic acid was added. Then the sample was vortexed again, capped, and heated at 100 °C for seven minutes. The sample was allowed to cool completely, then read at A820 and compared to a phosphorus standard curve to determine the total amount of phosphorus.

[0379] Example 11 - Liposome efflux assay

[0380] The data for this experiment are shown in Figure 2A .

[0381] A 16 mL liposome suspension was prepared with the following components: 160 μL of ferrozine solution (100X stock solution, 50 mM in the external liposome buffer), LUV suspension (to a final concentration of 1 mM phosphorus), and up to 16 mL of the external liposome buffer. Then 195 μL of the liposome suspension was added to a 96-well plate (NUM) using a multi-channel pipette. Then the plate was placed in a plate reader and equilibrated at 37 °C for 5 minutes, and then baseline measurements were taken. After equilibration, the compound at the specified concentration (40X stock solution in DMSO) was added using a multi-channel pipette, and then the plate was incubated at 37 °C for 40 minutes. Readings were taken every minute at A562 and the plate was mixed at 220 rpm. Total iron efflux was determined by the ferrocene-iron complex signal. These experiments were performed in triplicate with a minimum of two biological replicates.

[0382] Example 12 - H9C2 toxicity assay

[0383] The data for this experiment are shown in Figure 2B .

[0384] H9C2 cells (ATCC CRL-1446) were cultured in T175 culture flasks (Thermofisher) at 37 °C / 5% CO2 in DMEM medium supplemented with 10% FBS (Gemini 900-108), 1% penicillin-streptomycin (Gibco 15140-122), and 1% MEM non-essential amino acids (Corning 25-025-Cl) until at least passage 5. By centrifugation and dilution of the cell pellet, a 5×10 5 cells / mL solution was obtained. A 96-well compound plate containing 40x DMSO solutions of the required test compounds at concentrations exceeding the desired range was prepared using a 100 mM DMSO stock solution. Using a multi-channel micropipette, 3.34 μL of each stock solution was added to 297 μL of cell medium in the 96-well plate and gently mixed by pipetting up and down. To the 96-well tissue culture plate, 3 × 80 μL of each resulting compound solution was added to consecutive rows, and the top and bottom rows were filled with cell medium to reduce evaporation. Subsequently, 20 μL of the 5×10 5 cells / mL solution was added to each well using a micropipette such that each well contained 1×10 4 cells. The plate was incubated at 37 °C / 5% CO2 for four days, and then cell viability was measured using AlamarBlue reagent according to the assay protocol. Briefly, 10 μL of AlamarBlue reagent was added to each well, and then the plate was incubated for 3 hours. Cell viability was measured by fluorescence signal (Em555 / Ex585) using a microplate reader (BioTek Synergy H1 hybrid reader) and compared to puromycin- and DMSO-treated wells as positive and negative controls, respectively.

[0385] Example 13 - K562 toxicity assay

[0386] The data of this experiment are shown in Figure 2C .

[0387] K562 cells (ATCC CCL-243) were cultured in T175 culture flasks (Thermofisher) at 37 °C / 5% CO2 in IMDM (Iscove's Modified Dulbecco's Medium, ATCC 30-2005) medium supplemented with 10% FBS (Gemini 900-108), 1% penicillin-streptomycin (Gibco 15140-122), and 1% MEM non-essential amino acids (Corning 25-025-Cl) until at least passage 5. By centrifugation and dilution of the cell pellet, a 1.25×10 6cells / mL solution. Prepare a 96-well compound plate containing a 40x DMSO solution of the desired test compound at concentrations exceeding the desired range using a 100 mM DMSO stock solution. Using a multi-channel micropipette, add 3.34 μL of each stock solution to 297 μL of cell culture medium in the 96-well plate and mix gently by pipetting up and down. In the 96-well plate, add 3 × 80 μL of each resulting compound solution to consecutive rows, filling the top and bottom rows with cell culture medium to reduce evaporation. Subsequently, using a micropipette, add 20 μL of a 1.25×10 6 cells / mL solution to each well such that each well contains 2.5×10 4 cells. Incubate the plate at 37 °C / 5% CO2 for four days, then determine cell viability using the AlamarBlue reagent according to the assay protocol. Briefly, add 10 μL of AlamarBlue reagent to each well and then incubate the plate for 3 hours. Determine cell viability using a microplate reader (BioTek Synergy H1 hybrid reader) by fluorescence signal (Em555 / Ex585) and compare it to wells treated with puromycin and DMSO as positive and negative controls, respectively.

[0388] Example 14 - Procedure for Caenorhabditis elegans experiments

[0389] Data for this experiment are shown in Figure 3A-3H .

[0390] Caenorhabditis elegans strains:

[0391] BY200 vtIs1[pdat-1::gfp; rol-6]V

[0392] NES277 fpn-1.2-KOV; vtIs1[pdat-1::gfp; rol-6]V

[0393] GA631 wuIs177[ftn-1p::GFP+lin-15(+)]X

[0394] NES281 fpn-1.2-KOV; wuIs177[ftn-1p::gfp+lin-15(+)]X

[0395] Analysis of dopaminergic neurodegeneration

[0396] The BY200 and NES277 strains were grown on normal NGM plates. At 1-day-old, the worms were transferred to NGM plates containing vehicle (DMSO) or various concentrations of deferiprone (DFP), hinokitiol (Hino), and FeM-1269. The worms were passaged daily until imaged on day 7 using a Zeiss AxioImager microscope equipped with DIC and fluorescence. Blind data analysis was performed according to the previously described protocol (R. Nass, D. H. Hall, D. M. Miller, R. D. Blakely, Neurotoxin-induced degeneration of dopamine neurons in Caenorhabditis elegans. Proceedings of the National Academy of Sciences 99, 3264-3269 (2002).).

[0397] Measurement of ferritin fluorescence

[0398] The GA631 and NES281 strains were grown on normal NGM plates. At day 1, the worms were transferred to NGM plates containing vehicle (DMSO) or various concentrations of DFP, Hino, and FeM-1269. The worms were passaged until the heads were imaged at 20x magnification using a Zeiss LSM880 on day 5. Fluorescence intensity was blindly quantified using ImageJ.

[0399] Example 15 - Procedure for measuring midbrain iron levels in iron-overloaded mice

[0400] Data from this experiment are shown in Figure 4A -E.

[0401] Animal care

[0402] This study was conducted in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health (Bethesda, MD, USA). The protocol (protocol number: PRO00010742) was approved by the University of Michigan Committee on the Use and Care of Animals (UCUCA). The flatiron (ffe / +) mice were provided by I. E. Zohn (Children’s National Medical Center, Washington, DC). All mice used in these studies were on a 129S6 / SvEvTac background. Weaned mice were fed a diet (TD120518; Harlan Teklad, Indianapolis, IN, USA) as previously described (Y. A. Seo, J. A. Elkhader, M. Wessling-Resnick, Distribution of manganese and other biometals in flatiron mice. Biometals 29, 147-155 (2016); Y. A. Seo, M. Wessling-Resnick, Ferroportin deficiency impairs manganese metabolism in flatiron mice. FASEB J 29, 2726-2733 (2015)).

[0403] Elevated plus maze

[0404] This technique is widely used in mice to test anxiety-like behavior and locomotor activity (A. A. Walf, C. A. Frye, The use of the elevated plus maze as an assay of anxiety-related behavior in rodents. Nat Protoc 2, 322-328 (2007)). The elevated plus maze consists of two open arms and two closed arms extending from a central zone platform. Each mouse was placed in the central zone, facing an open arm, and allowed to explore the maze for 5 minutes. The time and frequency of entry into the three regions of the apparatus (i.e., open arms, closed arms, and central zone), as well as the total distance traveled, rearing frequency, and locomotor duration throughout the maze, were recorded and analyzed using EthoVision XT (Noldus).

[0405] Inductively coupled plasma mass spectrometry (ICP-MS) measurement of brain Fe levels

[0406] To characterize the acute and chronic effects of Hino on iron trafficking and distribution, iron mice and WT littermates were administered by intraperitoneal (IP) injection. Mice were euthanized at 4 hours or 1 week later, brains were collected, and iron content was determined by ICP-MS (Y.A. Seo, J.A. Elkhader, M. Wessling-Resnick, Distribution of manganese and other biometals in flatiron mice. Biometals 29, 147-155 (2016); Y.A. Seo, M. Wessling-Resnick, Ferroportin deficiency impairs manganese metabolism in flatiron mice. FASEB J 29, 2726-2733 (2015)).

[0407] Example 16 - Dynamic light scattering experiment

[0408] Dynamic light scattering was performed on a Malvern Zetasizer Nano using disposable cuvettes. Samples were evaluated in a buffer similar to the liposome efflux studies (50 mM MES / TRIS, pH adjusted to 7.0 using 1 M MES or 1 M TRIS solution). Compounds at the designated concentrations (1000X in DMSO) and equimolar FeCl3 (1000X in 0.1 M HCl) were added to this buffer. After addition, these compounds were vortexed for 30 seconds, sonicated for one minute, then vortexed for an additional 30 seconds, and then equilibrated for five minutes at 37 °C in the Zetasizer. All standard parameters were used and each plot represents a minimum of thirty 10-second measurements. For count rate analysis, the raw count rate was utilized, accounting for any differences in the decay sub-position between samples.

[0409] Incorporation by reference

[0410] All U.S. patents and U.S. and PCT patent application publications mentioned herein are hereby incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference in its entirety. In case of conflict, the present application (including any definitions herein) shall control.

[0411] Equivalent schemes

[0412] Although specific embodiments of the present invention have been discussed, the above specification is illustrative and not restrictive. After reading this specification and the appended claims, many variations of the present invention will become apparent to those skilled in the art. The full scope of the present invention should be determined by reference to the claims, together with the full scope of their equivalents, and this specification, together with such variations.

Claims

1. A compound of formula I: or a pharmaceutically acceptable salt thereof; wherein: R 1 is selected from the group consisting of: H, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl and -O-heterocycloalkyl; R 2 is selected from the group consisting of: H, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl and -O-heterocycloalkyl; and R 3 selected from the group consisting of: H, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl and -O-heterocycloalkyl; R 4 is selected from the group consisting of: H, C1-C5 alkyl, cycloalkyl, heterocycloalkyl, -O-C1-C5 alkyl, -O-cycloalkyl and -O-heterocycloalkyl; and R 5 selected from the group consisting of: H, a halogenated group, a C1-C5 alkyl group, a cycloalkyl group, a heterocycloalkyl group, -O-C1-C5 alkyl, -O-cycloalkyl and -O-heterocycloalkyl; Wherein when R 1 , R 2 , R 3 , R 4 or R 5 is a C1-C5 alkyl group, a cycloalkyl group, a heterocycloalkyl group, -O-C1-C5 alkyl group, -O-cycloalkyl group or -O-heterocycloalkyl group, at least one hydrogen atom in the C1-C5 alkyl group, cycloalkyl group, heterocycloalkyl group, -O-C1-C5 alkyl group, -O-cycloalkyl group or -O-heterocycloalkyl group is replaced by a deuterium atom; and wherein R 1 , R 2 , R 3 and R 4 at least one of which is not H.

2. The compound according to claim 1, wherein R 1 is selected from the group consisting of methyl, ethyl, propyl and butyl.

3. The compound according to claim 1, wherein R 1 is selected from the group consisting of CD3, C2D5, C3D7, and C4D9.

4. The compound according to claim 1, wherein R 1 is:

5. The compound according to claim 1, wherein R 1 is:

6. The compound according to claim 1, wherein R 1 is:

7. The compound according to claim 1, wherein R 1 is:

8. The compound according to claim 1, wherein R 1 is:

9. The compound according to claim 1, wherein R 1 is:

10. The compound according to claim 1, wherein R 1 is selected from the group consisting of: -OMe, -OEt, -OPr and -OBu.

11. The compound according to claim 1, wherein R 1 is selected from the group consisting of: -OCD3, -OC2D5, -OC3D7, and -OC4D9.

12. The compound according to claim 1, wherein R 1 is:

13. The compound according to claim 1, wherein R 1 is:

14. The compound according to any one of claims 1 to 13, wherein R 2 is selected from the group consisting of methyl, ethyl, propyl, and butyl.

15. The compound according to any one of claims 1 to 13, wherein R 2 is selected from the group consisting of CD3, C2D5, C3D7 and C4D9.

16. The compound according to any one of claims 1 to 13, wherein R 2 is:

17. The compound according to any one of claims 1 to 13, wherein R 2 is:

18. The compound according to any one of claims 1 to 13, wherein R 2 is:

19. The compound according to any one of claims 1 to 13, wherein R 2 is:

20. The compound according to any one of claims 1 to 13, wherein R 2 is:

21. The compound according to any one of claims 1 to 13, wherein R 2 is:

22. The compound according to any one of claims 1 to 13, wherein R 2 is selected from the group consisting of: -OMe, -OEt, -OPr, and -OBu.

23. The compound according to any one of claims 1 to 13, wherein R 2 is selected from the group consisting of: -OCD3, -OC2D5, -OC3D7 and -OC4D9.

24. The compound according to any one of claims 1 to 13, wherein R 2 is:

25. The compound according to any one of claims 1 to 13, wherein R 2 is:

26. The compound according to any one of claims 1 to 25, wherein R 3 is selected from the group consisting of methyl, ethyl, propyl and butyl.

27. The compound according to any one of claims 1 to 25, wherein R 3 is selected from the group consisting of CD3, C2D5, C3D7 and C4D9.

28. The compound according to any one of claims 1 to 25, wherein R 3 is:

29. The compound according to any one of claims 1 to 25, wherein R 3 is:

30. The compound according to any one of claims 1 to 25, wherein R 3 is:

31. The compound according to any one of claims 1 to 25, wherein R 3 is:

32. The compound according to any one of claims 1 to 25, wherein R 3 is:

33. The compound according to any one of claims 1 to 25, wherein R 3 is:

34. The compound according to any one of claims 1 to 25, wherein R 3 is selected from the group consisting of: -OMe, -OEt, -OPr, and -OBu.

35. A compound according to any one of claims 1 to 25, wherein R 3 is selected from the group consisting of: -OCD3, -OC2D5, -OC3D7, and -OC4D9.

36. The compound according to any one of claims 1 to 25, wherein R 3 is:

37. A compound according to any one of claims 1 to 25, wherein R 3 is:

38. A compound according to any one of claims 1 to 37, wherein R 4 is selected from the group consisting of methyl, ethyl, propyl and butyl.

39. A compound according to any one of claims 1 to 37, wherein R 4 is selected from the group consisting of CD3, C2D5, C3D7 and C4D9.

40. A compound according to any one of claims 1 to 37, wherein R 4 is:

41. A compound according to any one of claims 1 to 37, wherein R 4 is:

42. A compound according to any one of claims 1 to 37, wherein R 4 is:

43. A compound according to any one of claims 1 to 37, wherein R 4 is:

44. A compound according to any one of claims 1 to 37, wherein R 4 is:

45. A compound according to any one of claims 1 to 37, wherein R 4 is:

46. A compound according to any one of claims 1 to 37, wherein R 4 is selected from the group consisting of: -OMe, -OEt, -OPr, and -OBu.

47. A compound according to any one of claims 1 to 37, wherein R 4 is selected from the group consisting of: -OCD3, -OC2D5, -OC3D7 and -OC4D9.

48. A compound according to any one of claims 1 to 37, wherein R 4 is:

49. A compound according to any one of claims 1 to 37, wherein R 4 is:

50. A compound according to any one of claims 1 to 49, wherein R 5 is H.

51. The compound according to any one of claims 1 to 49, wherein R 5 is a halogenated group.

52. The compound according to any one of claims 1 to 49, wherein R 5 is F.

53. The compound according to any one of claims 1 to 49, wherein R 5 is selected from the group consisting of methyl, ethyl, propyl, and butyl.

54. A compound according to any one of claims 1 to 49, wherein R 5 is selected from the group consisting of CD3, C2D5, C3D7 and C4D9.

55. A compound according to any one of claims 1 to 49, wherein R 5 is:

56. A compound according to any one of claims 1 to 49, wherein R 5 is:

57. The compound according to any one of claims 1 to 49, wherein R 5 is:

58. A compound according to any one of claims 1 to 49, wherein R 5 is:

59. The compound according to any one of claims 1 to 49, wherein R 5 is:

60. The compound according to any one of claims 1 to 49, wherein R 5 is:

61. The compound according to any one of claims 1 to 49, wherein R 5 is selected from the group consisting of: -OMe, -OEt, -OPr, and -OBu.

62. The compound according to any one of claims 1 to 49, wherein R 5 is selected from the group consisting of: -OCD3, -OC2D5, -OC3D7, and -OC4D9.

63. The compound according to any one of claims 1 to 49, wherein R 5 is:

64. The compound according to any one of claims 1 to 49, wherein R 5 is:

65. The compound according to claim 1, wherein the compound is selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

66. The compound according to claim 1, wherein the compound is selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

67. The compound according to claim 1, wherein the compound is selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

68. A pharmaceutical composition comprising the compound according to any one of claims 1 to 67 or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient.

69. The pharmaceutical composition according to claim 68, which further comprises an additional therapeutic agent selected from the group consisting of an iron mobilizing agent and deferiprone.

70. A method of treating a disease or disorder characterized by reduced ferroportin, which comprises administering to a subject in need thereof a therapeutically effective amount of the compound according to any one of claims 1 to 67 or the pharmaceutical composition according to claim 68 or 69.

71. The method according to claim 70, wherein the disease or disorder characterized by reduced ferroportin is an age-related induced ferroportin disease.

72. A method of treating a disease or disorder characterized by reduced ferrotransporter, which comprises administering to a subject in need thereof a therapeutically effective amount of the compound according to any one of claims 1 to 67 or the pharmaceutical composition according to claim 68 or 69.

73. A method of treating a disease or disorder characterized by ferrotransporter defect or deficiency, which comprises administering to a subject in need thereof a therapeutically effective amount of the compound according to any one of claims 1 to 67 or the pharmaceutical composition according to claim 68 or 69.

74. A method of treating a disease or disorder characterized by iron accumulation, which comprises administering to a subject in need thereof a therapeutically effective amount of the compound according to any one of claims 1 to 67 or the pharmaceutical composition according to claim 68 or 69.

75. The method according to claim 74, wherein the iron accumulation is iron accumulation in the brain.

76. A method of treating a disease or disorder characterized by improper iron distribution, which comprises administering to a subject in need thereof a therapeutically effective amount of the compound according to any one of claims 1 to 67 or the pharmaceutical composition according to claim 68 or 69.

77. The method according to claim 76, wherein the improper iron distribution is improper iron distribution in the brain.

78. The method according to any one of claims 72 to 77, wherein the disease or disorder is selected from the group consisting of: inflammatory anemia, iron overload, thalassemia, hemochromatosis, atransferrinemia, myelodysplasia, hypochromic microcytic anemia, ferroportin disease, transfusion-induced iron overload, and aceruloplasminemia.

79. A method of treating a neurodegenerative disorder, which comprises administering to a subject in need thereof a therapeutically effective amount of the compound according to any one of claims 1 to 67 or the pharmaceutical composition according to claim 68 or 69.

80. The method according to claim 79, wherein the neurodegenerative disorder is selected from the group consisting of: Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, Friedrich's ataxia, Huntington's disease, Lewy body disease, and spinal muscular atrophy.

81. The method according to claim 79, wherein the neurodegenerative disorder is neurodegeneration with brain iron accumulation (NBIA).

82. The method according to claim 81, wherein the NBIA is selected from the group consisting of: beta-helix protein-related neurodegeneration (BPAN), pantothenate kinase-associated neurodegeneration (PKAN), PLA2G6-related neurodegeneration (PLAN), mitochondrial membrane protein-related neurodegeneration (MPAN), fatty acid hydroxylase-related neurodegeneration (FAHN), COASY protein-related neurodegeneration (CoPAN), aceruloplasminemia, Kufor-Rakeb syndrome (also known as Parkinson's disease 9 (PARK9)), neuroferritinopathy, Woodhouse-Sakati syndrome, and idiopathic NBIA.

83. The method according to any one of claims 70 to 82, further comprising administering to the subject an effective amount of an additional therapeutic agent selected from the group consisting of an iron mobilizing agent and deferiprone.

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