OA17479A - Compounds and uses thereof for the modulation of hemoglobin. - Google Patents

Abstract

Provide herein are compounds and pharmaceutical compositions suitable as modulators of hemoglobin, methods and intermediates for their preparation, and methods for their use in treating disorders mediated by hemoglobin and disorders that would benefit from tissue and/or cellular oxygenation.

Description

COMPOUNDS AND USES THEREOF FOR THE MODULATION OF HEMOGLOBIN

FIELD OFTHE INVENTION [0001] This invention provides compounds and pharmaceutical compositions suitable as allosteric modulators of hemoglobin, methods and intermediates for their préparation, and methods for their use in treating disorders mediated by hemoglobin and disorders that would benefit from tissue and/or cellular oxygénation.

STATE OF THE ART [0002] Sickle cell disease is a disorder of the red blood cells, found particularly among those of African and Mediterranean descent. The basis for sickle cell disease is found in

1Q sickle hemoglobin (HbS), which contains a point mutation relative to the prévalent peptide sequence of hemoglobin (H b).

[0003] Hemoglobin (Hb) transports oxygen molécules from the lungs to various tissues and organs throughout the body. Hemoglobin binds and releases oxygen through conformational changes. Sickle hemoglobin (HbS) contains a point mutation where glutamic acid is replaced with valine, allowing HbS to become susceptible to polymerization to give the HbS containing red blood cells their characteristic sickle shape. The sickled cells are also more rigid than normal red blood cells, and their lack of flexibility can lead to blockage of blood vessels. US 7,160,910 discloses compounds that are allosteric modulators of hemoglobin. However, a need exists for additional therapeutics that can treat disorders that are mediated by Hb or by abnormal Hb such as HbS.

SUMMARY OF THE INVENTION [0004] This invention relates generally to compounds and pharmaceutical compositions suitable as allosteric modulators of hemoglobin. In some aspects, this invention relates to methods for treating disorders mediated by hemoglobin and disorders that would benefit from tissue and/or cellular oxygénation.

[0005] In certain aspects of the invention, a compound of formula (I) is provided:

or a tautomer thereof, or a pharmaceutically acceptable sait of each thereof, wherein ring A is an optionally substituted 4-10 membered cycloalkyl or 4-10 membered heterocycle containing up to 5 ring heteroatoms, wherein the heteroatom is selected from the group consisting of O, N, S, and oxidized forms of N and S;

ring B is a C₆-Ci_Oaryl or 5-10 membered heteroaryl having 1-3 nitrogen atoms, preferably 1-2 nitrogen atoms and more preferably 1 nitrogen atom, or oxidized versions thereof, wherein the aryl or heteroaryl is optionally substituted;

/A

F' is a single or a double bond;

each Y and Z is independently CR¹⁰R¹¹,0, S, SO, SO2, or NR¹²; each R¹⁰ and R¹¹ independently is hydrogen or C1-C3 alkyl optionally substituted with halo, OH, or Ci-C₆ alkoxy, or CR¹⁰Rⁿ is C=O; R¹² is hydrogen or Ci-Ce alkyl; provided that if one of Y and Z is O, S, SO, SO2, then the other is not CO, and provided that Y and Z are both not heteroatoms or oxidized forms thereof;

ring C is C₅-Cio aryl, optionally substituted;

V¹ and V² independently are Ci-C₆ alkoxy; or V¹ and V² together with the carbon atom they are attached to form a ring of formula:

wherein each V³ and V⁴ are independently O, S, or N H, provided that when one of V³ and V⁴ is S, the other is NH, and provided that V³ and V⁴ are both not NH; q is 1 or 2; each V⁵ is independently Ci-Cô alkyl or CO₂R⁶⁰, where each R⁶⁰ independently is Ci-K alkyl or hydrogen; t is 0,1, 2, or 4; or CV³V² is C=V, wherein V is O, NOR⁸⁰, or NNR⁸¹R⁸²;

R⁸⁰ is optionally substituted Ci-Ce alkyl;

R⁸¹ and R⁸² independently are selected from the group consisting of hydrogen, optionally substituted Ci-C6 alkyl, COR⁸³, or CO2R⁸⁴;

R⁸³ is hydrogen or optionally substituted Ci-C₆ alkyl; and

R⁸⁴ is optionally substituted Ci-Cg alkyl.

[0006] In certain aspects of the invention, a compound of formula (IA) is provided:

(IA) wherein R⁵ is hydrogen, Ci-C₆ alkyl or a prodrug moiety R, wherein the Ci-C₆ alkyl is optionally substituted with 1-5 halo;

R⁶ is halo, Ci-C₆ alkyl, Ci-Ce alkoxy, Ci-Cg alkylthio, Ci-C₆ S(O)-, Ci-C₆ S(O)₂-,wherein the Ci-Ce alkyl is optionally substituted with 1-5 halo; or

R⁶ is 4-10 membered cycloalkyl or heterocycle substituted with an R'R'N- moiety wherein each R' is independently Ci-Cô alkyl or hydrogen;

k is 0 or 1;

p is 0,1, 2 or 3;

and the remaining variables are defined as above.

[0007] In further aspects of the invention, a composition is provided comprising any of the compounds described herein, and at least a pharmaceutically acceptable excipient.

[0008] In still further aspects of the invention, a method is provided for increasing oxygen affinity of hemoglobin S in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of any of the compounds or compositions described herein.

[0009] In further aspects of the invention, a method is provided for treating oxygen deficiency associated with sickle cell anémia, the method comprising administering to a subject in need thereof a therapeutically effective amount of any of the compounds or compositions described herein.

DETAILED DESCRIPTION OFTHE INVENTION

Définitions [0010] It must be noted that as used herein and in the appended claims, the singular forms a, an, and the include plural referents unless the context clearly dictâtes otherwise. Thus, for example, reference to a solvent includes a plurality of such solvents.

[0011] As used herein, the term comprising or comprises is intended to mean that the compositions and methods include the recited éléments, but not exduding others. Consisting essentially of when used to define compositions and methods, shall mean exduding other éléments of any essential significance to the combination for the stated purpose. Thus, a composition or process consisting essentially ofthe éléments as defined herein would not exdude other materials or steps that do not materially affect the basic and novel characteristic(s) ofthe claimed invention. Consisting of shall mean exduding more than trace éléments of other ingrédients and substantial method steps. Embodiments defined by each of these transition terms are within the scope of this invention.

[0012] Unless otherwise indicated, ali numbers expressing quantities of ingrédients, reaction conditions, and so forth used in the spécification and claims are to be understood as being modified in ali instances by the term about. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following spécification and attached claims are approximations. Each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. The term about when used before a numerical désignation, e.g., température, time, amount, and concentration, including range, îndicates approximations which may vary by ( + ) or ( - ) 10 %, 5 % or 1 %.

[0013] As used herein, C_m-C_n, such as C1-C12, Ci-C₈, or Ci-C₆ when used before a group refers to that group containing m to n carbon atoms.

[0014] The term alkoxy refers to -O-alkyl. The term alkylthio is -S-alkyl.

[0015] The term alkyl refers to monovalent saturated aliphatic hydrocarbyl groups having from 1 to 30 carbon atoms (i.e., Ci-C₃₀ alkyl) or 1 to 22 carbon atoms (i.e., C1-C22 alkyl), 1 to 8 carbon atoms (i.e., Ci-C₈ alkyl), or 1 to 4 carbon atoms. This term includes, by way of example, linear and branched hydrocarbyl groups such as methyl (CH₃-), ethyl (CH3CH2-), n-propyl (CH₃CH₂CH2-), isopropyl ((CH₃)₂CH ), n-butyl (CH3CH2CH2CH2-), isobutyl ((CHahCHCHz-), sec-butyl ((CH₃)(CH₃CH₂)CH-), t-butyl ((CH₃)₃C-), n-pentyl (CH3CH2CH2CH2CH2), and neopentyl ((CHshCCFh-).

[0016] The term aryl refers to a monovalent, aromatic mono- or bicyclic ring having 6-10 ring carbon atoms. Examples of aryl include phenyl and naphthyl. The condensed ring may or may not be aromatic provided that the point of attachment is at an aromatic carbon atom. For example, and without limitation, the following is an aryl group:

[0017] The term -CO2H ester refers to an ester formed between the -CO2H group and an alcohol, preferably an aliphatic alcohol. A preferred example included -CÛ2R^E, wherein R^E is alkyl or aryl group optionally substituted with an amino group.

[0018] The term chiral moiety refers to a moiety that is chiral. Such a moiety can possess one or more asymmetric centers. Preferably, the chiral moiety is enantiomerically enriched, and more preferably a single enantiomer. Non limiting examples of chiral moieties include chiral carboxylic acids, chiral amines, chiral amino acids, such as the naturally occurring amino acids, chiral alcohols including chiral steroids, and the likes.

[0019] The term cycloalkyl refers to a monovalent, preferably saturated, hydrocarbyl mono-, bi-, or tricyclic ring having 3-12 ring carbon atoms. While cycloalkyl, refers preferably to saturated hydrocarbyl rings, as used herein, it also includes rings containing 12 carbon-carbon double bonds. Nonlimiting examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamentyl, and the like. The condensed rings may or may not be non-aromatic hydrocarbyl rings provided that the point of attachment is at a cycloalkyl carbon atom. For example, and without limitation, the following is a cycloalkyl group:

[0020] The term halo refers to F, Cl, Br, and/or I.

[0021] The term heteroaryl refers to a monovalent, aromatic mono-, bi-, or tricyclic ring having 2-16 ring carbon atoms and 1-8 ring heteroatoms selected preferably from N, O, S, and P and oxidized forms of N, S, and P, provided that the ring contains at least 5 ring atoms. Nonlimiting examples of heteroaryl include furan, imidazole, oxadiazole, oxazole, pyridine, quinoline, and the like. The condensed rings may or may not be a heteroatom containing aromatic ring provided that the point of attachment is a heteroaryl atom. For example, and without limitation, the following is a heteroaryl group:

[0022] The term heterocyclyl or heterocycle refers to a non-aromatic, mono-, bi-, or tricyclic ring containing 2-12 ring carbon atoms and 1-8 ring heteroatoms selected preferably from N, O, S, and P and oxidized forms of N, S, and P, provided that the ring contains at least 3 ring atoms. While heterocyclyl preferably refers to saturated ring Systems, it also includes ring Systems containing 1-3 double bonds, provided that the ring is non-aromatic. Nonlimiting examples of heterocyclyl include, azalactones, oxazoline, piperidinyl, piperazinyl, pyrrolidinyl, tetrahydrofuranyl, and tetrahydropyranyl. The condensed rings may or may not contain a non-aromatic heteroatom containing ring provided that the point of attachment is a heterocyclyl group. For example, and without limitation, the following is a heterocyclyl group:

[0023] The term hydrolyzing refers to breaking an R^H-O-CO-, R^H-O-CS-, or an R^H-O-SO₂moiety to an R^h-OH, preferably by adding water across the broken bond. A hydrolyzing is performed using various methods well known to the skilled artisan, non limiting examples of which include acidic and basic hydrolysis.

[0024] The term oxo refers to a C=O group, and to a substitution of 2 geminal hydrogen atoms with a C=O group.

[0025] The term optionally substituted refers to a substituted or unsubstituted group. The group may be substituted with one or more substituents, such as e.g., 1, 2, 3, 4 or 5 substituents. Preferably, the substituents are selected from the group consisting of oxo, halo, -CN, NO₂, -N₂+, -CO₂R¹⁰⁰, -OR¹⁰⁰, -SR¹⁰⁰, -SOR¹⁰⁰, -SO2R¹⁰⁰, -NR¹⁰¹R¹⁰², -CONR¹⁰¹R¹⁰², SO2NR¹⁰¹R¹⁰², Ci-C6 alkyl, Ci-C6 alkoxy, -CR¹⁰⁰=C(R¹⁰⁰)2, -CCR¹⁰⁰, C3-Ci0 cycloalkyl, C3-C10 heterocyclyl, C6-Ci2aryl and C2-Ci2 heteroaryl, wherein each R¹⁰⁰ independently is hydrogen or Ci-C8 alkyl; C₃-C_i2 cycloalkyl; C3-C10 heterocyclyl; Ce-Cuaryl; or C₂-Ci₂ heteroaryl; wherein each alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with 1-3 halo, 1-3 Ci-Cg alkyl, 1-3 C1-C6 haloalkyl or 1-3 CiA alkoxy groups. Preferably, the substituents are selected from the group consisting of chloro, fluoro, -OCH3, methyl, ethyl, fco-propyl, cyclopropyl, vinyl, ethynyl, -CO₂H, -CO₂CH3, -OCF3, -CF3 and -OCHF₂.

[0026] R¹⁰¹ and R¹⁰² independently is hydrogen; Ci-C8 alkyl, optionally substituted with CO2H or an ester thereof, Ci-Ce alkoxy, oxo, -CR¹⁰³=C(R¹⁰³)2, -CCR, C3-C10 cycloalkyl, C3-C10 heterocyclyl, C6-Ci₂aryl, or C₂-Ci₂ heteroaryl, wherein each R¹⁰³ independently is hydrogen or Ci-C₈ alkyl; C3-C12 cycloalkyl; C3-C10 heterocyclyl; C6-C_i2 aryl; or C₂-Ci₂ heteroaryl; wherein each cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with 1-3 alkyl groups or 1-3 halo groups, or R¹⁰¹ and R¹⁰² together with the nitrogen atom they are attached to form a 5-7 membered heterocycle.

[0027] The term pharmaceutically acceptable refers to safe and non-toxic for in vivo, preferably, human administration.

[0028] The term pharmaceutically acceptable sait refers to a sait that is pharmaceutically acceptable.

[0029] The term sait refers to an ionic compound formed between an acid and a base.

When the compound provided herein contains an acidic functionality, such salts include, without limitation, alkali métal, alkaline earth métal, and ammonium salts. As used herein, ammonium salts include, salts containing protonated nitrogen bases and alkylated nitrogen bases. Exemplary, and non-limiting cations useful in pharmaceutically acceptable salts include Na, K, Rb, Cs, NH₄, Ca, Ba, imidazolium, and ammonium cations based on naturally occurring amino acids. When the compounds utilized herein contain basic functionality, such salts include, without limitation, salts of organic acids, such as carboxylic acids and sulfonic acids, and minerai acids, such as hydrogen halides, sulfuric acid, phosphoric acid, and the likes. Exemplary and non-limiting anions useful in pharmaceutically acceptable salts include oxalate, maleate, acetate, propionate, succinate, tartrate, chloride, sulfate, bisalfate, mono-, di-, and tribasic phosphate, mesylate, tosylate, and the likes.

[0030] The terms treat, treating or treatment, as used herein, include alleviating, abating or ameliorating a disease or condition or one or more symptoms thereof, preventing additional symptoms, ameliorating or preventing the underlying metabolic causes of

2q symptoms, inhibiting the disease or condition, e.g., arresting or suppressing the development of the disease or condition, relieving the disease or condition, causing régression of the disease or condition, relieving a condition caused by the disease or condition, or suppressing the symptoms of the disease or condition, and are intended to include prophylaxis. The terms also include relieving the disease or conditions, e.g., causing the régression of clinical symptoms. The terms further include achieving a therapeutic benefit and/or a prophylactic benefit. By therapeutic benefit is meant éradication or amelioration of the underlying disorder being treated. Also, a therapeutic benefit is achieved with the éradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the individual, notwithstanding that the individual is still be afflicted with the underlying disorder. For prophylactic benefit, the compositions are administered to an individual at

risk of developing a particular disease, or to an individual reporting one or more of the physiological symptoms of a disease, even though a diagnosis of this disease has not been made.

[0031] The terms preventing or prévention refer to a réduction in risk of acquiring a disease or disorder (i.e., causing at least one of the clinical symptoms of the disease not to develop in a subject that may be exposed to or predisposed to the disease but does not yet expérience or display symptoms ofthe disease). The terms further include causing the clinical symptoms not to develop, for example in a subject at risk of suffering from such a disease ordisorder, thereby substantially averting onset ofthe disease or disorder.

[0032] The term effective amount refers to an amount that is effective for the treatment of a condition or disorder by an intranasal administration of a compound or composition described herein. In some embodiments, an effective amount of any of the compositions or dosage forms described herein is the amount used to treat a disorder mediated by hemoglobin or a disorder that would benefit from tissue and/or cellular oxygénation of any ofthe compositions or dosage forms described herein to a subject in need thereof.

[0033] The term carrier as used herein, refers to relatively nontoxic chemical compounds or agents that facilitate the incorporation of a compound into cells, e.g., red blood cells, or tissues.

[0034] As used herein, a prodrug is a compound that, after administration, is metabolized or otherwise converted to an active or more active form with respect to at least one property. To produce a prodrug, a pharmaceutically active compound can be modified chemically to render it less active or inactive, but the chemical modification is such that an active form ofthe compound is generated by metabolic or other biological processes. A prodrug may hâve, relative to the drug, altered metabolic stability or transport characteristics, fewer side effects or lower toxicity. For example, see the reference Nogrady, 1985, Médicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392. Prodrugs can also be prepared using compounds that are not drugs.

Compounds [0035] In certain aspects of the invention, a compound of formula (I) is provided:

(D or a tautomer thereof, or a pharmaceutically acceptable sait of each thereof, wherein ring A is an optionally substituted 4-10 membered cycloalkyl or 4-10 membered heterocycle containing up to 5 ring heteroatoms, wherein the heteroatom is selected from the group consisting of O, N, S, and oxidized forms of N and S;

ring B is a C₆-Ci₀aryl or 5-10 membered heteroaryl having 1-3 nitrogen atoms, preferably 1-2 nitrogen atoms and more preferably 1 nitrogen atom, or oxidized versions thereof, wherein the aryl or heteroaryl is optionally substituted;

each Y and Z is independently CR¹⁰Rⁿ, O, S, SO, SO2, or NR¹²; each R¹⁰ and R¹¹ independently is hydrogen or C1-C3 alkyl optionally substituted with halo, OH, or Cj-Cg alkoxy, or CR^1OR^U is C=O; R¹² is hydrogen or CpCg alkyl; provided that if one of Y and Z is O, S, SO, SO₂, then the other is not CO, and provided that Y and Z are both not heteroatoms or oxidized forms thereof;

ring Cis C₆-Ci₀aryl;

V¹ and V² independently are Ci-Ce alkoxy; or V¹ and V² together with the carbon atom they are attached to form a ring of formula:

wherein each V³ and V⁴ are independently O, S, or NH, provided that when one of V³ and V⁴ is S, the other is NH, and provided that V³ and V⁴ are both not NH; q is 1 or 2; each V⁵ is independently Ci-Cg alkyi or CO2R⁶⁰, where each R⁶⁰ independently is Ci-Ce alkyi or hydrogen; t is 0,1, 2, or 4; or CV^XV² is C=V, wherein V is O, NOR⁸⁰, or NNR⁸¹R⁸²;

R⁵ is hydrogen, Ci-C_s alkyi or a prodrug moiety R, wherein the Ci-Cg alkyi is optionally substituted with 1-5 halo;

R⁶ is halo, Ci-C₆ alkyi, Ci-C₆ alkoxy, Ci-Cg alkylthio, Ci-C₆ S(O)-, Ci-C₆ S(O)₂-,wherein the C1-C6 alkyi is optionally substituted with 1-5 halo; or

R⁶ is 4-10 membered cycloalkyl or heterocycle substituted with an R'R'N- moiety wherein each R' is independently C1-C6 alkyi or hydrogen;

R⁸⁰ is optionally substituted Ci-Ce alkyi;

R⁸¹ and R⁸² independently are selected from the group consisting of hydrogen, optionally substituted Ci-C6 alkyi, COR⁸³, or CO2R⁸⁴;

R⁸³ is hydrogen or optionally substituted C1-C6 alkyi;

R⁸⁴ is optionally substituted Ci-Ce alkyi;

k is 0 or 1; and p is 0,1, 2 or 3.

[0036] In certain embodiments, t is 0. In certain embodiments, t is 1. In certain embodiments, t is 2. In certain embodiments, t is 3.

[0037] Preferably, in certain embodiments, Y and Z are both not a heteroatom or a heteroatom containing moiety. Preferably, one of Y and Z is a methylene or substituted methylene and the other is a heteroatom or a heteroatom containing moiety. More preferably, Y is an alkylene, and Z is a heteroatom or a heteroatom containing moiety, which, yet more preferably is oxygen.

[0038] Preferably, V¹ and V² together with the carbon atom they are attached to form a ring of formula:

[0039] In some embodiments, V¹ and V² independently are Ci-C₆ alkoxy; or V¹ and V² together with the carbon atom they are attached to form a ring of formula:

wherein each V³ and V⁴ are independently O, S, or NH, provided that when one of V³ and V⁴ is S the other is N H, and provided that V³ and V⁴ are both not NH; q is 1 or 2; each V⁵ is independently Ci-Ce alkyl or CO2R⁶⁰, where each R⁶⁰ independently is Ci-Ce alkyl or hydrogen; t is 0,1, 2, or 4; or CVN² is C=V, wherein V is O, and wherein the remaining variables are defined herein.

[0040] In certain aspects ofthe invention, the compound of Formula (I) is of Formula (II):

wherein the remaining variables are defined herein.

[0041] In certain aspects of the invention, the compound of Formula (I) is of Formula (11A):

wherein the variables are defined herein.

[0042] In some embodiments, ring A is optionally substituted with 1-3: halo, Ci-Cg alkyl, COR¹⁵ and/or COOR¹⁵; wherein R^1S is optionally substituted Ci-C₆ alkyl, optionally substituted C6-Ci₀aryl, optionally substituted 5-10 membered heteroaryl containing up to 5 ring heteroatoms, or optionally substituted 4-10 membered heterocycle containing up to 5 ring heteroatoms, wherein the heteroatom is selected from the group consisting of O, N, S, and oxidized forms of N and S.

[0043] In some embodiments, ring B is optionally substituted with 1-3: halo, Ci-C₆ alkyl COR¹⁵ and/or COOR¹⁵; wherein R¹⁵ is optionally substituted Ci-Cg alkyl, optionally substituted Ce-Cioaryl, optionally substituted 5-10 membered heteroaryl containing up to 5 ring heteroatoms, or optionally substituted 4-10 membered heterocycle containing up to 5 ring heteroatoms, wherein the heteroatom is selected from the group consisting of O, N, S, and oxidized forms of N and S.

[0044] In some embodiments, the compound is selected from the group consisting of

or an N oxide thereof, wherein

R¹⁴ is Ci-C₆ alkyl, C₃-C₈ cycloalkyl, COR¹⁵ or COOR¹⁵;

R¹⁵ is optionally substituted Ci-Q alkyl, optionally substituted C₆-Ci₀aryl, optionally substituted 5-10 membered heteroaryl containîng up to 5 ring heteroatoms, or optionally substituted 4-10 membered heterocycle containîng up to 5 ring heteroatoms, wherein the heteroatom is selected from the group consisting of O, N, S, and oxidized forms of N and S;

x is 0,1, or 2;

p is 0,1, and 2; and m is 0,1 or 2.

[0045] In one embodiment, the compound is

or a prodrug thereof, or a pharmaceutidaly acceptable sait of each thereof.

[0046] Other compounds provided herein are included in the Examples section.

Prodrug Moiety [0047] In one aspect, R is hydrogen, a phosphate or a diphosphate containing moiety, or ₅ another promoiety or prodrug moiety. Preferably the prodrug moiety imparts at least a 2 fold, more preferably a 4 fold, enhanced solubility and/or bioavailability to the active moiety (where R is hydrogen), and more preferably is hydrolyzed in vivo. The promoieties are structurally and functionally defined herein.

[0048] In one embodiments, R is -COR⁹⁰, CO₂R⁹¹, or CONR⁹²R⁹³ wherein

R⁹⁰ and R⁹¹ independently are Ci-Ce alkyl, C₃-C₈ cycloalkyl, 4-9 membered heterocycle, or a

5-10 membered heteroaryl, each containing at least 1 basic nitrogen moiety; and

R⁹² and R⁹³ independently are Ci-Cg alkyl; C₃-C₈ cycloalkyl, 4-9 membered heterocycle, or a 510 membered heteroaryl, each containing at least 1 basic nitrogen moiety; or R⁹² and R⁹³

together with the nitrogen atom they are bonded to for a 4-9 member heterocycle substituted with at least 1 amino, Ci-C₆ alkyl amino, or di Ci-Cg alkylamino group.

[0049] In certain embodiments, R is -C(O)R³¹, C(O)OR³¹, or CON(R¹³)2, each R³¹ is independently a C1-C6 alkyl; C3-C₈ cycloalkyl, 4-9 membered heterocycle, or a 5-10 membered heteroaryl, containing at least 1 basic nitrogen moiety; and each R¹³ independently are Ci-C6 alkyl; C3-C8 cycloalkyl, 4-9 membered heterocycle, or a 5-10 membered heteroaryl, containing at least 1 basic nitrogen moiety; or 2 R¹³ together with the nitrogen atom they are bonded to for a 4-9 member heterocycle substituted with at least 1 amino, Ci-C6 alkyl amino, or di Ci-Ce alkylamino group.

[0050] In one aspect, R is C(O)OR³¹, C(S)OR³¹, C(O)SR³¹ or COR³¹, wherein R³¹ is as defined herein.

[0051] In one embodiment, R³¹ is a group of the formula (CR³²R³³)_eNR³⁴R³⁵, wherein each R³² and R³³ is independently H, a Ci-C8 alkyl, C3-C9 heterocyclyl, C3-C8 cycloalkyl, C6-C10 aryl, C3-C9 heteroaryl or R³² and R³³ together with the carbon atom they are bond to form a C3-C8 cycloalkyl, Ce-Cio aryl, C₃-Cg heterocyclyl or C3-C9 heteroaryl ring system, or 2 adjacent R³² moieties or 2 adjacent R³³ moieties together with the carbon atom they are bond to form a C₃-C₈ cycloalkyl, C₆-Ci₀ aryl, C3-C9 heterocyclyl or C3-C9 heteroaryl ring system;

each R³⁴ and R³⁵ is a Ci-C8 alkyl, C3-C9 heterocyclyl, C3-C8 cycloalkyl, or R³⁴ and R³⁵ together with the nitrogen atom they are bond to form a C3-C₈ cycloalkyl or C3-C9 heterocyclyl ring system;

each heterocyclic and heteroaryl ring system is optionally substituted with C1-C3 alkyl, -OH, amino and carboxyl groups; and e is an integer of from 1 to 4.

[0052] In some less preferred embodiments R³⁴ and R³⁵ can be hydrogen.

[0053] In one embodiment, the subscript e is preferably 2 and each R³² and R³³ is preferably independently selected from the group, H, CH₃, and a member in which R³² and R³³ are joined together to form a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or l,l-dioxo-hexahydro-IA⁶-thiopyran-4-yl or tetrahydropyran-4-yl group.

[0054] With regard to the prodrug group, preferred embodiments are compounds wherein NR³⁴R³⁵ is morpholino.

[0055] In one embodiment, R is:

wherein each R³² and R³³ is independently H, Ci-C₈ alkyl, or optionally, if both présent on the same substituent, may be joined together to form a C3-C8 cycloalkyl, C₆-Ci₀ aryl, C3-C9 heterocyclyl or C3-C9 heteroaryl ring system.

[0056] Within this embodiment, each R³² and R³³ is independently, H, CH3, or are joined together to form a cyclopropyl, cyclopbutyl, cyclopentyl, cyclohexyl, 1,1-dioxo- hexahydroIX⁶-thiopyran-4-yl or tetrahydropyran-4-yl group.

[0057] In a preferred embodiment, linkage ofthe prodrug moiety to the rest ofthe active molécule isstable enough sothat the sérum half life ofthe prodrug isfrom about8 to about 24 hours.

[0058] In an embodiment of the invention, the prodrug moiety comprises a tertiary amine having a pKa near the physiological pH of 7.5. Any amines having a pKa within 1 unit of 7.5 are suitable alternatives amines for this purpose. The amine may be provided by the amine of a morpholino group. This pKa range of 6.5 to 8.5 allows for significant concentrations of the basic neutral amine to be présent in the mildly alkaline small intestine. The basic, neutral form ofthe amine prodrug is lipophilie and is absorbed through the wall ofthe small intestine into the blood. Following absorption into the bloodstream, the prodrug moiety is cleaved by esterases which are naturally présent in the sérum to release an active compound.

[0059] Examples of R include, without limitation:

[0060] In another embodiment, R is as tabulated below:

R	m	R34	R35	nr34r35
C(O)(CH2)mNR34R35	1	Me	Me
C(O)(CH2)mNR34R35	2	Me	Me
C(O)(CH2)mNR34R35	3	Me	Me
C(O)(CH2)mNR34R35	4	Me	Me
C(O)(CH2)mNR34R35	1
C(O)(CH2)mNR34R35	2
C(O)(CH2)mNR34R35	3
C(O)(CH2)mNR34R35	4			A-/
C(O)O(CH2)mNR34R35	2	Me	Me
C(O)O(CH2)mNR34R35	3	Me	Me
C(O)O(CH2)mNR34R35	4	Me	Me

C(O)O(CH2)mNR34R35	2
C(O)O(CH2)mNR34R35	3
C(O)O(CH2)mNR34R35	4			A-/
P(O)(OH)2
an N oxide thereof, or a pharmaceutically acceptab	e sait of each thereof.

[0061] In another aspect, R is,

wherein

R³⁶ is lower alkyl (e.g. Ci-C₆ alkyl).

[0062] In yet another aspect, R is:

wherein X¹, Y¹ and X² are as defined herein.

[0063] In one embodiment, X¹ is selected from the group consisting of O, S and NR³⁷ wherein R³⁷ is hydrogen or Ci-Ce alkyl;

Y¹ is -C(R³⁸)2 or a sugar moiety, wherein each R³⁸ is independently hydrogen or Ci-C5 alkyl, C₃-C₈ cycloalkyl, C3-C9 heterocyclyl, Ce-Cio aryl, or C3-C9 heteroaryl;

X² is selected from the group consisting of halogen, Ci-Ce alkoxy, diacylglycérol, amino, Ci-C₆ alkylamino, C1-C6 dialkylamino, Ci-C₆ alkylthio, a PEG moiety, a bile acid moiety, a sugar moiety, an amino acid moiety, a di-or tri-peptide, a PEG carboxylic acid, and -U-V wherein

U is O or S; and

V is selected from the group consisting of Ci-Ce alkyl, C3-C8 cycloalkyl, C3-C9 heterocyclyl, C6-C10 aryl, C3-C9 heteroaryl, C(W²)X³, PO(X³)₂, and SO₂X³;

wherein W² is O or NR³⁹ wherein R³⁹ is hydrogen or Ci-Ce alkyl, C3-C8 cycloalkyl, C3-C9 hetrocyclyl, C₆-Ci₀ aryl, or C3-C9 heteroaryl; and each X³ is independently amino, hydroxyl, mercapto, Ci-C6 alkyl, heteroalkyl, cycloalkyl, hetrocyclyl, aryl, or heteroaryl, C1-C6 alkoxy, Ci-Cg alkylamino, Ci-C6 dialkylamino, Ci-Cô alkylthio, a bile acid based alkoxy group, a sugar moiety, a PEG moiety, and -O-CH2-CH(OR⁴⁰)CH2X⁴R⁴⁰, wherein:

X⁴ is selected from the group consisting of O, S, S=O, and SO₂; and each R⁴⁰ is independently Ci₀-C₂₂ alkyl, C₃-C₈ cycloalkyl, C3-C9 heterocyclyl, C₆-Ci₀ aryl, or C3-C9 heteroaryl, Ci-C₈ alkylene, or Ci-C₈ heteroalkylene.

[0064] Each heterocyclic and heteroaryl ring system is optionally substituted with C1-C3 alkyl, -OH, amino and carboxyl groups.

[0065] In one embodiment, the présent invention utilizes the following Y¹ groups: CH2, CHMe, CH(isopropyl), CH(tertiarybutyl), C(Me)2, C(Et)2, C(isopropyl)2, and C(propyl)2.

[0066] In another embodiment, the présent invention utilizes the following X² groups:

-OMe, -OEt, -O-isopropyl, O-isobutyl, O-tertiarybutyl, -O-COMe, -O-C(=O)(isopropyl), -O-C(=O)(isobutyl), -O-C(=O)(tertiarybutyl), -O-C(=O)-NMe₂, -O-C(=O)-NHMe, -O-C(=O)-NH₂, -O-C(=O)-N(H)-CH(R⁴¹)-CO2Et wherein R⁴¹ is a side chain Ci-C₆ alkyl, or C3-C9 heterocyclyl group selected from the side chain groups présent in essential amino acids; -O-P(=O)(OMe)2, -O-P(=O)(O-isopropyl)₂, and -0-P(=0)(0-isobutyl)₂. Each heterocyclic is optionally substituted with one or more, preferably, 1-3, C1-C3 alkyl, -OH, amino and/or carboxyl groups.

[0067] In another embodiment, In one embodiment, R is:

wherein

X³ is independently C1-C6 alkyl, C₃-C₈ cycloalkyl, C3-C9 heterocyclyl, C₆-Ci₀ aryl, or C3Cg heteroaryl; and

R⁴² is independently hydrogen or Οχ-Οβ alkyl, C3-C8 cycloalkyl, C3-C9 heterocyclyl, Cg-Cio aryl, or C3-C9 heteroaryl.

[0068] Each heterocyclic is optionally substituted with one or more, preferably, 1-3, C1-C3 alkyl, -OH, amino and/or carboxyl groups.

[0069] In one embodiment, R is:

,or

wherein j each X³ is independently amino, hydroxyl, mercapto, Ci-Ce alkyl, C3-C8 cycloalkyl, C3Cg heterocyclyl, C6-Ci0 aryl, or C3-C9 heteroaryl, Ci-C6 alkoxy, Ci-Cg alkylamino, Ci-C6 dialkylamino, Ci-Ce alkylthïo, a bile acid based alkoxy group, a sugar moiety, a PEG moiety, and -O-CH2-CH(OR⁴⁰)CH2X⁴R⁴⁰, wherein:

ΊΟ X⁴ is selected from the group consisting of O, S, S=O, and SO₂; and each R⁴⁰ is independently Ci₀-C₂₂ alkyl, C₃-C₈ cycloalkyl, C3-C9 heterocyclyl, C6-C10 aryl, C3-C9 heteroaryl, Ci-C₈ alkylene, or Ci-C₈ heteroalkylene; and

R⁴² is independently hydrogen or Ci-C₆ alkyl, C₃-C₈ cycloalkyl, C3-C9 heterocyclyl, Cg-Cio aryl, or C3-C9 heteroaryl.

5 [0070] In some embodiments, R⁴² is independently hydrogen or Ci-C₆ alkyl, C₃-C₈ cycloalkyl, C3-C9 heterocyclyl, Ce-Cio aryl, or C3-C9 heteroaryl; and each X³ independently is Ci-C₆ alkyl, C₃-C₈ cycloalkyl, C3-C9 heterocyclyl, C₆-Ci₀ aryl, or C3-C9 heteroaryl, C1-C6 alkoxy, Ci-C₆ alkylamino, Ci~C₆ dialkylamino, or Ci-C₆ alkylthïo.

[0071] In some embodiments, R is represented by the following structures:

wherein, in the above examples, R⁴³ is Ci₀-C₂2 alkyl or alkylene, R⁴⁴ is H or Cx-Cô alkyl and R⁴⁵ represents side chain alkyl groups présent in naturally occurring alpha amino acids;

wherein R⁴⁶ is (CH2)n, f=2-4, and CO-R⁴⁷-NH2 represents an aminoacyl group; or

wherein R⁴⁶ is (CH2)n, n=2-4, R⁴⁷ is (CH2)_n, n=l-3 and R⁴⁹ is O or N Me.

[0072] ln one embodiment, R is:

O

A [0073] In one aspect, R is -C(R²⁰⁰R²⁰¹)O(R²⁰²R²⁰³)P(O)OR²⁰⁴NR²⁰⁵R²⁰⁶, wherein each R²⁰⁰, . R²⁰¹, R²⁰², R²⁰³, R²⁰⁴ R²⁰⁵ and R²⁰® is independently H, a Ci-C₈ alkyl, C3-C9 heterocyclyl, C3-C8 cycloalkyl, C₆-Ci₀ aryl, C3-C9 heteroaryl, wherein each alkyl, heterocyclyl, cycloalkyl, aryl, and heteroaryl is optionally substituted.

[0074] In some embodiments, R is -CH(R²⁰¹)OCH2P(O)OR²⁰⁴NHR²⁰⁶, wherein R²⁰¹ is CrC₈ alkyl, R²⁰⁴is phenyl, optionally substituted. In one embodiment, R²⁰⁶ is -CHR²⁰⁷C(O)OR²⁰⁸ wherein R²⁰⁷ is selected from the group consisting of the naturally occurring amino acid side chains and CO₂H esters thereof and R²⁰⁸ is Ci-C8 alkyl. In one embodiment, R²⁰⁶ is Ci-Cg alkyl, optionally susbtitued with 1-3, CO2H, SH, NH^ C₆-Cio aryl, and C₂-C₁₀ heteroaryl.

[0075] In some embodiments, R is:

[0076] In one embodiment, R is:

wherein Y¹ is -CiR³⁸^, wherein each R³⁸ is independently hydrogen or Ci-C₆ alkyl, C₃C₈ cycloalkyl, C3-C9 heterocyclyl, C₆-Ci₀ aryl, or C3-C9 heteroaryl.

[0077] Various polyethylene glycol (PEG) moieties and synthetic methods related to them that can be used or adapted to make compounds ofthe invention are described in U.S.

Patent Nos. 6,608,076; 6,395,266; 6,194,580; 6,153,655; 6,127,355; 6,111,107; 5,965,566;

5,880,131; 5,840,900; 6,011,042 and 5,681,567.

wherein

R⁵⁰ is -OH or hydrogen;

R⁵¹ is -OH, or hydrogen;

W is- CHfCHslW¹;

wherein W¹ is a substituted Ci-C₈ alkyl group containing a moiety which is optionally negatively charged at physiological pH, said moiety is selected from the group consisting of CO₂H, SO₃H, SO₂H, -P(O)(OR⁵²)(OH), -OP(O)(OR⁵²)(OH), and OSO₃H, wherein R⁵² is Ci-C₆ alkyl, C₃-C₈ cycloalkyl, C₃-C₉ heterocyclyl, C₆-Ci₀ aryl, or C3-C9 heteroaryl.

[0079] Each heterocyclic and heteroaryl ring system is optionally substituted with one or more, preferably 1-3, C1-C3 alkyl, -OH, amino and/or carboxyl groups.

[0080] In one embodiment, R is:

wherein R⁵³ is H or Ci-C₆ alkyl.

[0081] In another aspect, R isSOsH.

[0082] In another aspect, R comprises a cleavable linker, wherein the term cleavable linker refers to a linker which has a short half life in vivo. The breakdown of the linker Z in a compound releases or generates the active compound. In one embodiment, the cleavable linker has a half life of less than ten hours. In one embodiment, the cleavable linker has a half life of lessthan an hour. In one embodiment, the half life ofthe cleavable linkeris between one and fifteen minutes. In one embodiment, the cleavable linker has at least one connection with the structure: C*- C(=X*)X*-C* wherein C* is a substituted or unsubstituted methylene group, and X* is S or O. In one embodiment, the cleavable linker has at least one C*-C(=O)O-C* connection. In one embodiment, the cleavable linker has at least one C*C(=O)S-C* connection. In one embodiment, the cleavable linker has at least one -C(=O)N*C*-SO₂-N*-connection, wherein N* is -NH- or Ci-Cg alkylamino. In one embodiment, the cleavable linker is hydrolyzed by an esterase enzyme.

[0083] In one embodiment, the linker is a self-immolating linker, such as that disclosed in U.S. patent publication 2002/0147138, to Firestone; PCT Appl. No. US05/08161 and PCT Pub. No. 2004/087075. In another embodiment, the linker is a substrate for enzymes. See generally Rooseboom et al., 2004, Pharmacol. Rev. 56:53-102.

Pharmaceutical Compositions [0084] In further aspects of the invention, a composition is provided comprising any of the compounds described herein, and at least a pharmaceutically acceptable excipient.

q [0085] In another aspect, this invention provides a composition comprising any of the compounds described herein, and a pharmaceutically acceptable excipient.

[0086] Such compositions can be formulated for different routes of administration. Although compositions suitable for oral delivery will probably be used most frequently, other routes that may be used include transdermal, intravenous, intraarterial, pulmonary, 25 rectal, nasal, vaginal, lingual, intramuscular, intraperitoneal, intracutaneous, intracranial, and subcutaneous routes. Suitable dosage formsforadministering any ofthe compounds described herein include tablets, capsules, pills, powders, aérosols, suppositories, parenterals, and oral liquids, încluding suspensions, solutions and émulsions. Sustained release dosage forms may also be used, for example, in a transdermal patch form. Ail

dosage forms may be prepared using methods that are standard in the art (see e.g., Remington's Pharmaceutical Sciences, 16^th ed., A. Oslo editor, Easton Pa. 1980).

[0087] Pharmaceutically acceptable excipients are non-toxic, aid administration, and do not adversely affect the therapeutic benefit of the compound of this invention. Such excipients may be any solid, liquid, semi-solid or, in the case of an aérosol composition, gaseous excipient that is generally available to one of skill in the art. Pharmaceutical compositions in accordance with the invention are prepared by conventional means using methods known in the art.

[0088] The compositions disclosed herein may be used in conjunction with any of the vehicles and excipients commonly employed in pharmaceutical préparations, e.g., talc, gum arable, lactose, starch, magnésium stéarate, cocoa butter, aqueous or non-aqueous solvents, oils, paraffin dérivatives, glycols, etc. Coloring and flavoring agents may also be added to préparations, particularly to those for oral administration. Solutions can be prepared using water or physiologically compatible organic solvents such as éthanol, 1,2propylene glycol, polyglycols, dimethylsulfoxide, fatty alcohols, triglycérides, partial esters of glycerin and the like.

[0089] Solid pharmaceutical excipients include starch, cellulose, hydroxypropyl cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnésium stéarate, sodium stéarate, glycerol monostearate, sodium chloride, dried skim milk and the like. Liquid and semisolid excipients may be selected from glycerol, propylene glycol, water, éthanol and various oils, including those of petroleum, animal, vegetable or synthetic origin, e.g., peanut oil, soybean oil, minerai oil, sesame oil, etc. In certain embodiments, the compositions provided herein comprises one or more of α-tocopherol, gum arable, and/or hydroxypropyl cellulose.

[0090] In one embodiment, this invention provides sustained release formulations such as drug depots or patches comprising an effective amount of a compound provided herein. In another embodiment, the patch further comprises gum Arabie or hydroxypropyl cellulose separately or in combination, in the presence of alpha-tocopherol. Preferably, the hydroxypropyl cellulose has an average MW of from 10,000 to 100,000. In a more preferred embodiment, the hydroxypropyl cellulose has an average MW of from 5,000 to 50,000.

[0091] Compounds and pharmaceutical compositions of this invention maybe used alone or in combination with other compounds. When administered with another agent, the coadministration can be in any manner in which the pharmacological effects of both are manifest in the patient at the same time. Thus, co-administration does not require that a single pharmaceutical composition, the same dosage form, or even the same route of administration be used for administration of both the compound of this invention and the other agent or that the two agents be administered at precisely the same time. However, co-administration will be accomplished most conveniently by the same dosage form and the same route of administration, at substantially the same time. Obviously, such administration most advantageously proceeds by delivering both active ingrédients simultaneously in a novel pharmaceutical composition in accordance with the présent invention.

Methods of Treatment [0092] In aspects of the invention, a method is provided for increasing tissue and/or cellular oxygénation, the method comprising administering to a subject in need thereof a therapeutically effective amount of any of the compounds or compositions described herein.

[0093] In aspects of the invention, a method is provided for increasing oxygen affinity of hemoglobin S in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of any of the compounds or compositions described herein.

[0094] In aspects of the invention, a method is provided for treating a condition associated with oxygen deficiency, the method comprising administering to a subject in need thereof a therapeutically effective amount of any of the compounds or compositions described herein.

[0095] In further aspects of the invention, a method is provided for treating oxygen deficiency associated with sickle cell anémia, the method comprising administering to a subject in need thereof a therapeutically effective amount of any of the compounds or compositions described herein.

[0096] In further aspects of the invention, a method is provided for treating sickle cell disease, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any ofthe compounds or compositions described herein. In still further aspects of the invention, a method is provided for treating cancer, a pulmonary disorder, stroke, high altitude sickness, an ulcer, a pressure sore, Alzheimer's disease, acute respiratory disease syndrome, and a wound, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any ofthe compounds or compositions described herein.

Synthetic Methods [0097] Certain methods for making the compounds described herein are also provided. The reactions are preferably carried out in a suitable inert solvent that will be apparent to the skilled artisan upon reading this disclosure, for a sufficient period of time to ensure substantial completion of the reaction as observed by thin layer chromatography, ’H-NMR, etc. If needed to speed up the reaction, the reaction mixture can be heated, as is well known to the skilled artisan. The final and the intermediate compounds are purified, if necessary, by various art known methods such as crystallization, précipitation, column chromatography, and the likes, as will be apparent to the skilled artisan upon reading this disclosure.

[0098] An illustrative and non-limiting method for synthesizing a compound of formula (I), is schematically shown below.

herein.

X and X⁵ each represent a leaving group and are independently selected from Cl, F, Br, and I.

X⁶ represents CHR¹⁴, NR¹⁴, O, S(O)x ; wherein x is 0,1, or 2;

Y⁵ represents a leaving group selected from Cl, F, Br, I, OSO2R¹⁷ and OSO2Ar;

R¹⁷ is Ci-Ce alkyl;

n is 0,1, or 2;

Ar is phenyl optionally substituted with 1-3 halo or C1-C4 alkyl groups.

Where variables already used in the structures hereinabove are used in the shcemes, the context makes it unambiguous as to what the variable refers to.

General Synthetic Scheme IA

4a or 4b [0099] General method A for preparing aryloxy ether analogs (4a) from substituted methylene alcohol (1) and hydroxyl aryl aldéhyde dérivative (3a). A hydroxyl arylaldehyde dérivative (3a) (0.1-2 mmol) mixture with substituted methylene alcohol (1) (0.8 to 1.2eq) and PPhî (1-1.5eq) in anhydrous THF (1-lOmL) was stirred under nitrogen until complété dissolution. The solution was cooled to 0 °C on ice bath and DIAD or DEAD (1.1 eq) in THF or toluene was added dropwise over a 1-20 min period. The ice cooling bath was allowed to expire over 90 min and the mixture was stirred at RT for 2-48 hours. The mixture was stirred for 10 min, then filtered through a pad of silica. The silica was washed with ethyl acetate 220mL. The combined filtrâtes were evaporated and the residue was dried on highvac. The residue was purified by préparative HPLC or flash silica gel chromatography.

[0100] General method B for preparing aryloxy ether analogs (4a) from substituted methylene halide (2) and hydroxyl aryl aldéhyde dérivatives (3a). A mixture of hydroxyl arylaldehyde dérivatives (3a) (0.1-2 mmol, 1-4 eq.), substituted methylene chloride or bromide (2) (leq), and K₂CO₃ (2-5 eq.) (catalytic amount of Nal or Bu₄NI may also be added) in DMF or acetonitrile (1 to 10 mL) was stirred at RT or heating up to 120 °C for 0.5-8 h

under nitrogen atmosphère. In workup A, water was added to the reaction mixture, the precipitated product was collected, washed with water, and then subjected to préparative HPLC or flash silica gel chromatography purification. In workup B (for products that did not precipitate), diluted HCl or aqueous NH4CI was added at 0 °C to adjusted the pH to ~7, the reaction mixture was partitioned between ethyl acetate or dichloromethane and aqueous sodium chloride and the organic layer separated, dried, and solvent removed under vacuum to afford crude product which was purified by automated silica gel column chromatography using appropriate solvents mixture (e.g., ethyl acetate/hexanes).

[0101] General method C for preparing substituted methylene chloride (2a). To a solution of substituted methylene alcohol (1) (0.1 to 2 mmol) in DCM (1-10 mL) was added SOCI2 dropwise (2eq to 5eq ) at 0 °C or RT. The reaction mixture was stirred at RT for 10min to 6 h, or until reaction is judged complété (LC/MS). The reaction mixture is concentrated to dryness over a rotavap. The crude chloride residue was suspended in toluene, sonicated and concentrated to dryness. The process was repeated three times and dried under vacuum to give the substituted methylene chloride (2), usually as an off-white solid, which was used for next step without further purification. Altematively, a solution of aqueous IN Na₂CO₃ is then added to produce a solution of pH~ 8. the mixture was extracted with DCM (3 xl0-50mL), dried over sodium sulfate, and concentrated to the crude substituted methylene chloride (2a), which is then purified by column chromatography on silica gel (0100% ethyl acetate-hexanes).

[0102] General method D for preparing substituted methylene bromide (2b). To a solution of substituted methylene alcohol (1) (0.1 to 2 mmol) in DCM (1-10 mL) was added Ph₃PBr₂ dropwise (2eq to 5eq ) at 0 °C or RT. The reaction mixture was stirred at RT for 10 min to 2 h, or until reaction is judged complété (LC/MS). The reaction mixture is concentrated to dryness over a rotavap. The residue purified by column chromatography on silica gel (0-100% ethyl acetate-hexanes) to afford the pure bromide 2b.

[0103] Similarly, N-linked heterocyclic analogs (compound 5) can also be synthesized from amination procedures developed by Buchwald and Hartwig.

Scheme IB

X

[0104] Protected amides of formula -CONHR⁹⁵ and -CONHOR⁹⁵ can be converted e.g., hydrolyzed to the corresponding amides according to methods known to the skilled artisan.

C19H23N3O3: 342.2.

[0105] General Synthetic Scheme 2 (six membered ring)

	Stepl	Step2	Step3		Step4
	A Or . CFOR, fA'B	n2O .a	Ar- B(OR)2	<a'b _	LAH or DIBAL
V 0	j? ^οοσ-^γ1 FACTOR, O	RiOÎX/^γ OTf		Ar		rV OH Ar
5	R1=M6. Et 6	7		8,		9-OH
				Step6 I WC		| Step5
A. B = CH2,	CHR, CR2. NR, O. S, S(O)n	0	Step7
		RiOOC^YY At 11-trans		- Ar 11-cis		X? X Ar
		|	Step8	I		10-x
		Y OH Ar		X? OH Ar
		12-OH-tran	s	12-OH-cis
		|	Slep9	I
		X?		Y
		X Ar		X Ar
		13-X-trarrs		13-X-cis

[0106] General method E (Scheme 2) for preparing heterocyclic methylene dérivatives 9,

10,12 and 13. Condensation of heterocyclic ketone analog 5 with chlorformate or dialkyl carbonate gives (hetero)cyclic beta-ketone ester 6 (Step 1). The ketone ester 6 is converted

to the triflate intermediate 7 by treating with a triflating agent (e.g, triflic anhydride) in the presence of an organic base such as Hunig's base (Step 2). Suzuki coupling of the triflate 7 with a boronic acid or ester affords heterocyclohexene carboxylate 8 (Step 3). Subséquent réduction of the ester group by LAH or DIBAL gives the corresponding alcohol 9-OH (Step 4).

Further reaction of the alcohol 9-OH with thionyl chloride, Ph₃PBr₂ (or CBr₄-Ph3P or PBr₃), or alkyl/aryl sufonyl chloride produces the corresponding 10-X chloride, bromide or sulfonate (Step 5).

[0107] Altematively, the double bond of heterocyclohexene carboxylate 8 is reduced to give the c/s-heterocyclohexane 11-cis carboxylate under palladium catalyzed hydrogénation ₁ θ conditions (Step 6). Réduction of the ester group of 11-cis by LAH or DIBAL yields cis-alcohol

12-OH-cis (Step 8). Conversion of the alcohol 12-OH-cis to its chloride, bromide or sulfonate (such as mesylate, tosylate) 13-X-cis can be achieved by reacting with thionyl chloride, or Ph3PBr₂, or sufonyl chloride (such as mesyl chloride or tosyl chloride) (Step 9). The ciscyclohexane carboxylate 11-cis can also be isomerized to the thermodynamically more stable trans-isomer 11-trans by the treatment with an alcoholic alkoxide (e.g., ethoxide) solution. Analogously, transformation of 11-trans ester to 12-trans alcohol and 13-X-trans halide is accomplished by applying conditions of Step 8 and Step9 (Scheme 2) similar to these for the corresponding cis-isomers.

Scheme 3

q <

> I

Scheme 1 !

12-OH-CiS 12-OH-trana

Method B

13-X-cis

13-X-trans

III

! Q i / i^{2 x} • Scheme 1 [0108] Coupling of the (hetero)cyclic methylene dérivatives 9,10,12 and 13 with hydroxyl (hetero)arylaldehyde dérivatives (3a/3b) (see, e.g., Scheme 3) by general method A or B affords the corresponding aryloxy/heteroarylether analogs (4c and 4d).

Y= halide, OTs, OMs; OH [0109] Step la-Compound 13 can is synthesized via O-alkylation of phénol aldéhyde 12 with alkyl halide 11 (Y=halide, OTs, OMs). A mixture of hydroxyl (hetero)arylaldehyde dérivatives (12) (0.1-2 mmol, 1-4 eq.), substituted methylene chloride or bromide (11) (leq), and K₂CO₃ (2-5 eq.) (catalytic amount of Nal or Bu₄NI may also be added) in DMF, acetonitrile, NMP or DMSO (1 to 10 mL) was stirred at RT or heating up to 120 °C for 1-24 h under nitrogen atmosphère. In workup A, water was added to the reaction mixture, the precipitated product was collected, washed with water, and then subjected to préparative HPLC or flash silica gel chromatography purification. In workup B (for products that did not precipitate), diluted HCl or aqueous NH4CI was added at 0 °C to adjusted the pH to ~7, the reaction mixture was partitioned between ethyl acetate or dichloromethane and aqueous sodium chloride and the organic layer separated, dried, and solvent removed under vacuum to afford crude product which was purified by automated silica gel column chromatography using appropriate solvents mixture (e.g., ethyl acetate/hexanes).

[0110] Step lb-Alternatively, compound 13 is made by coupîing of phénol aldéhyde 12 with alcohol 1 (Y=OH) under Mitsunobu conditions. A hydroxyl (hetero)arylaldehyde dérivatives (12) (0.1-2 mmol) mixture with substituted methylene alcohol (11, Y=OH) (0.8 to 1.2eq) and (polymer-supported)/PPh3 (l-1.5eq) in anhydrous THF (1-lOmL) was stirred under nitrogen until complété dissolution. The solution was cooled to 0 °C on ice bath and DIAD or DEAD (1.1 eq) in THF or toluene was added drop wise over a 1-20 min period. The ice cooling bath was allowed to expire over 90 min and the mixture was stirred at RT for 248 hours. The mixture was stirred for 10 min, then filtered through a pad of silica. The silica was washed with ethyl acetate 2-20mL. The combined filtrâtes were evaporated and the residue was dried on highvac. The residue was purified by préparative HPLC or flash silica gel chromatography.

[0111] Step 2. To a solution of (2-chloropyridin-3-yl)nnethanol or (2-bromopyridin-3yl)methanol (l-100mmol) and appreciate bronic acid or ester (0.8 to 1.5 eq) in dioxane (2200 mL) was added a solution of sodium bicarbonate (3 eq) in water (1-100 mL), followed by the addition of Pd(dppf)CI2 (5 to 10mol%). After heating at 100 °C for 4-24 h, the reaction mixture was cooled and diluted with EtOAc, organic layer was washed with water, brine, dried and concentrated to give crude product, which was purified by column chromatography.

General Synthetic Scheme 5

COOR1

A = CH, N X= F, Cl, Br, B=CR2, NR,

[0112] Compound 25 can be prepared from 2-halonicotinate through a sériés organic transformations that involve displacement with cyclic amine and réduction of ester to give hydroxymethylene dérivative 22 (step 1). The final product can be synthesized via either direct Mitsunobu reaction of 22 with phénol aldéhyde 24 or conversion of the alcohol 22 to halide 23 followed by O-alkylation of phénol 24 with 23.

Prodrug Synthesis [0113] Synthèses of the ester prodrugs start with the free carboxylic acid bearing the tertiary amine. The free acid is activated for ester formation in an aprotic solvent and then reacted with a free alcohol group in the presence of an inert base, such as triethyl amine, to provide the ester prodrug. Activating conditions for the carboxylic acid include forming the acid chloride using oxalyl chloride or thionyl chloride in an aprotic solvent, optionally with a catalytic amount of dimethyl formamide, followed by évaporation. Examples of aprotic

solvents, include, but are not limited to methylene chloride, tetrahydrofuran, and the like. Alternatively, activations can be performed in situ by using reagents such as BOP (benzotriazol-l-yloxytris(dimethylamino) phosphonium hexafluorolphosphate, and the like (see Nagy et al., 1993, Proc. Natl. Acad. Sci. USA 90:6373-6376) followed by reaction with the free alcohol. Isolation of the ester products can be affected by extraction with an organic solvent, such as ethyl acetate or methylene chloride, against a mildly acidic aqueous solution; followed by base treatment of the acidic aqueous phase so as to render it basic; followed by extraction with an organic solvent, for example ethyl acetate or methylene chroride; évaporation of the organic solvent layer; and recrystalization from a solvent, such as éthanol. Optionally, the solvent can be acidified with an acid, such as HCl or acetic acid to provide a pharmaceutically acceptable sait thereof. Alternatively the crude reaction can be passed over an ion exchange column bearing sulfonic acid groups in the protonated form, washed with deionized water, and eluted with aqueous ammonia; followed by évaporation.

[0114] Suitable free acids bearing the tertiary amine are commercially available, such as 2(N-morpholino)-propionic acid, N,N- dimethyl-beta-alanine, and the like. Non- commercial acids can be synthesized in straightforward manner via standard literature procedures.

[0115] Carbonate and carbamate prodrugs can be prepared in an analogous way. For example, amino alcohols and diamines can be activated using activating agents such as phosgene or carbonyl diimidazole, to provide an activated carbonates, which in turn can react with the alcohol and/or the phenolic hydroxy group on the compounds utilized herein to provide carbonate and carbamate prodrugs.

[0116] Various protecting groups and synthetic methods related to them that can be used or adapted to make compounds of the invention can be adapted from the references Testa et al., Hydrolysis in Drug and Prodrug Metabolism, June 2003, Wiley- VCH, Zurich, 419-534 and Beaumont et al., Curr. Drug Metab. 2003,4:461-85.

[0117] Provided herein is a method of synthesizing an acyloxymethyl version of a prodrug by adapting a method from the reference Sobolev et al., 2002, J. Org. Chem. 67:401-410.

R⁵¹ is Ci-Cô alkyl.

[0118] Provided herein is a method for synthesizing a phosphonooxymethyl version of a prodrug by adapting a method from Mantyla et al., 2004, J. Med. Chem. 47:188-195.

NaH, DMF tetrabutylammonium bromide Q14 NaH, THF

YoEt

OEt [0119] Provided herein is a method of synthesizing an alkyloxymethyl version of a prodrug

R⁵² is Ci-C₆ alkyl, C₃-C₈ cycloalkyl, C3-C9 heterocyclyl, Ce-Cio aryl, or C₃-C₉ heteroaryl.

Examples [0120] The following examples are given for the purpose of illustrating various embodiments ofthe invention and are not meantto limitthe présent invention in any fashion. The présent examples, along with the methods described herein are presently représentative of preferred embodiments, are exemplary, and are not intended as limitations on the scope ofthe invention. Changes therein and other uses which are encompassed within the spirit ofthe invention as defined bythe scope ofthe claimswill occur to those skilled in the art.

[0121] In the examples below as well as throughout the application, the following abbreviations hâve the following meanings. If not defined, the terms hâve their generally accepted meanings.

°C = degrees Celsius

RT = Room température min = minute(s) h = hour(s) pL = Microliter mL = Milliliter mmol = Millimole eq = Equivalent mg = Milligram

MS = Mass spectrometry

LC-MS = Liquid chromatography-mass spectrometry

HPLC = High performance liquid chromatography

NMR = Nuclear magnetic résonance

EtOAc = Ethyl acetate

Ph₃PBr₂ = Triphenylphosphine dibromide

DMF = N, N-Dimethylformamide

DCM = Dichloromethane

DMSO = Dimethyl sulfoxide

THF = Tetrahydrofuran

DIAD = Diisopropyl azodicarboxylate

DEAD = Diethyl azodicarboxylate [0122] Préparation of 2-[[2-[(3R)-3-fluoropyrrolidin-l-yl]pyridin-3-yl]methoxy]-6hydroxybenzaldehyde

[0123] Step 1: (R)-ethyl 2-(3-fluoropyrrolidin-l-yl)nicotinate. To a solution of ethyl 2fluoronicotinate (0.074 g, 0.48 mmol) in DMF (0.3 mL) was added diisopropylethyl amine (0.25 mL, 1.4 mmol), and (R)-3-fIuoropyrrolidine (0.090 g, 0.72 mmol). The resulting mixture was irradiated with microwaves (100 °C) for lh and loaded directly onto a silica column.

Eluting the column with EtOAc/hexanes (0-100%) provided (R)-ethyl 2-(3-fluoropyrrolidin-lyl)nicotinate as a clear oil (0.100 g, 94% yield); MS (ESI) m/z 239 [M+H]⁺.

[0124] Step 2: (R)-(2-(3-fluoropyrrolidin-l-yl)pyridin-3-yl)methanol. To a cooled (0 °C) solution of (R)-methyl 2-(3-fluoropyrrolidin-l-yl)nicotinate in THF (5 mL) was added a solution of lithium aluminum hydride (IM in THF). The reaction mixture was stirred for lh 1Q and then 20 pL of H₂O was added followed by 20 pL of 15% NaOH (aq) and then 60 pL of additional water. The slurry was stirred for lh and filtered and the resulting residue was washed with ether. The combined organic layers were dried over MgSO₄ and concentrated in vacuo. Purification by column chromotography (EtOAc/hexanes,0-100%) provided (R)-(2(3-fluoropyrrolidin-l-yl)pyridin-3-yl)methanol (0.081 g, 92% yield). MS (ESI) m/z 197 [M+H]⁺.

₁₅ [0125] Step 3: (R)-3-(chloromethyl)-2-(3-fluoropyrrolidin-l-yl)pyridine. To a cooled (0 °C) solution of (fi)-(2-(3-fluoropyrrolidin-l-yl)pyridin-3-yl)methanol (0.081 g, 0.38 mmol) in dichloromethane was added SOCI₂ (0.450 g, 3.8 mmol) and the reaction mixture was allowed to warm to ambient température. After 1 h, the reaction mixture was concentrated and azeotroped with toluene to provide (/?)-3-(chloromethyl)-2-(3-fluoropyrrolidin-l20 yl)pyridine (0.080 g, 92%) as a clear oil. MS (ESI) m/z 215 [M+H]⁺.

[0126] Step 4: (R)-2-((2-(3-fluoropyrrolidin-l-yl)pyridin-3-yl)methoxy)-6hydroxybenzaldehyde. To a solution of (R)-3-(chloromethyl)-2-(3-fluoropyrrolidin-lyl)pyridine (0.080 g, 0.35 mmol) and 2,6-dihydroxybenzaldehyde (0.130 g, 0.94 mmol) in DMF was added potassium carbonate (0.190 g, 1.4 mmol) and the reaction mixture was

heated (60 °C). After 30 minutes, the DMF was removed and the resulting residue was reconstituted in CH₂CI₂ and filtered through a plug of silica (EtOAc/hexanes, 1:1). Purification Prep-HPLC provided (R)-2-((2-(3-fluoropyrrolidin-l-yl)pyridin-3-yl)methoxy)-6hydroxybenzaldehyde ( 8 mg, 5% yield). *H NMR (400 MHz, OMSO-cfe) δ 11.71 (dd, J= 8.4,

0.7 Hz, 1H), 10.21 (d, J = 0.5 Hz, 1H), 8.10 (dd, J = 4.8,1.9 Hz, 1H), 7.71 (dd, J = 7.4,1.9 Hz,

1H), 7.52 (t, J = 8.4 Hz, 1H), 6.73 (dd, J = 8.6, 0.7 Hz, 1H), 6.71 (dd, J = 7.4, 5.0 Hz, 1H), 6.53 (dt, J = 8.4, 0.7 Hz, 1H), 5.40 (dd, J = 54.2, 3.3 Hz, 1H), 5.28 (d, J = 11.3 Hz, 1H), 5.17 (d, J = 12.0 Hz, 1H), 3.91 - 3.56 (m, 4H), 2.21 -1.93 (m, 2H); MS (ESI) m/z 317 [M+H]⁺.

[0127] GBT883-(/î)-2-((2-(3-fluoropyrrolidin-l-yl)pyridin-3-yl)methoxy)-6hydroxybenzaldehyde. The compound was prepared from ethyl 2-fluoronicotinate and (/?)3-fluoropyrrolidine according to scheme 5, reaction steps 1, 3 and 4.

[0128] Step la: To a solution of ethyl 2-fluoronicotinate (0.074 g, 0.48 mmol) in DMF (0.3 mL) was added diisopropylethyl amine (0.25 mL, 1.4 mmol), and (R)-3-fluoropyrrolidine (0.090 g, 0.72 mmol). The resulting mixture was irradiated with microwaves (100 °C) for lh and loaded directly onto a silica column. Eluting the column with EtOAc/hexanes (0-100%) provided (/?)-ethyl 2-(3-fluoropyrrolidin-l-yl)nicotinate as a clear oil (0.100 g, 94% yield). MS (ES) for C_nH₁₅FN₂O₂: 225 (MH⁺).

[0129] Step lb: To a cooled (0 °C) solution of (/?)-methyl 2-(3-fluoropyrrolidin-lyl)nicotinate in THF (5 mL) was added a solution of lithium aluminum hydride (IM in THF). The reaction mixture was stirred for lh and then 20 pL of H2O was added followed by 20 pL of 15% NaOH (aq) and then 60 pL of additional water. The slurry was stirred for lh and filtered and the resulting residue was washed with ether. The combined organic layers were dried over MgSÜ4 and concentrated in vacuo. Purification by column chromotography (EtOAc/hexanes,0-100%) provided (/î)-(2-(3-fluoropyrrolidin-l-yl)pyridin-3-yl)methanol (0.081 g, 92% yield). MS (ES) for Ci₀Hi₃FN₂O:197 (MH⁺).

[0130] Step 3: To a cooled (0 °C) solution of (fi)-(2-(3-fluoropyrrolidin-l-yl)pyridin-3yl)methanol (0.081 g, 0.38 mmol) in dichloromethane was added SOCI₂ (0.450 g, 3.8 mmol) and the reaction mixture was allowed to warm to ambient température. After 1 h, the reaction mixture was concentrated and azeotroped with toluene to provide (/?)-3(chloromethyl)-2-(3-fluoropyrrolidin-l-yl)pyridine (0.080 g, 92%) as a clear oil. MS (ES) for C₁₀Hi₂CIFN₂: 215 (MH⁺).

[0131] Step 4. To a solution of (R)-3-(chloromethyl)-2-(3-fluoropyrrolidin-l-yl)pyridine (0.080 g, 0.35 mmol) and 2,6-dihydroxybenzaldehyde (0.130 g, 0.94 mmol) in DMF was added potassium carbonate (0.190 g, 1.4 mmol) and the réaction mixture was heated (60 °C). After 30 minutes, the DMF was removed and the resulting residue was reconstituted in CH₂CI₂ and filtered through a plug of silica (EtOAc/hexanes, 1:1). Purification Prep-HPLC provided (/?)-2-((2-(3-fluoropyrrolidin-l-yl)pyridin-3-yl)methoxy)-6-hydroxybenzaldehyde ( 8 mg, 5% yield). ^XH NMR (400 MHz, DMSO-cfg) δ 11.71 (dd, J = 8.4, 0.7 Hz, 1H), 10.21 (d, J = 0.5 Hz, 1H), 8.10 (dd, J = 4.8, 1.9 Hz, 1H), 7.71 (dd, J = 7.4, 1.9 Hz, 1H), 7.52 (t, J = 8.4 Hz, 1H), 6.73 (dd, J = 8.6, 0.7 Hz, 1H), 6.71 (dd, J = 7.4, 5.0 Hz, 1H), 6.53 (dt, J = 8.4, 0.7 Hz, 1H), 5.40 (dd, J = 54.2, 3.3 Hz, 1H), 5.28 (d, J = 11.3 Hz, 1H), 5.17 (d, J = 12.0 Hz, 1H), 3.91 - 3.56 (m, 4H), 2.21 -1.93 (m, 2H). MS (ES) for C₁₇Hi₇FN₂O₃: 317 (MH⁺).

GBT910

[0132] GBT910-2-((2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)pyridin-3-yl)methoxy)-6hydroxybenzaldehyde. The compound was prepared from ethyl 2-fluoronicotinate and 8oxa-3-azabicyclo[3.2.1]octane according to reaction scheme below.

Step 1a

[0133] Step la:To a solution of ethyl 2-fluoronicotinate (0.15 g, 0.97 mmol) in NMP (0.5 mL) was added diisopropylethyl amine (0.50 mL, 2.9 mmol), and 8-oxa-3azabicyclo[3.2.1]octane (0.17 g, 0.72 mmol). The resulting mixture was irradiated with microwaves (100 °C) for lh and loaded directly onto a silica column. Eluting the column with EtOAc/hexanes (0-100%) provided methyl 2-(8-oxa-3-azabicyclo[3.2.1]octan-3yl)nicotinate as a clear oil (0.100 g, 42% yield). MS (ES) for C13H16N2O3: 249 (MH⁺).

[0134] Step lb: To a cooled (0 °C) solution of 2-(8-oxa-3-azabicyclo[3.2.1]octan-3yljnicotinate (0.10 g, 0.40 mmol) in THF (5 mL) was added a solution of lithium aluminum hydride (1.2 mL, IM in THF). The reaction mixture was stirred for lh and then 20 pL of H₂O was added followed by 20 pL of 15% NaOH (aq) and then 60 pL of additional H₂O. The slurry was stirred for lh, filtered and the resulting residue was washed with ether. The combined organic layers were dried over MgSO₄ and concentrated in vacuo to yield (2-(8-oxa-3azabicyclo[3.2.1]octan-3-yl)pyridin-3-yl)methanol (0.070 g, 79% yield). MS (ES) for C12H16N2O2: 221 (MH⁺).

[0135] Step 2: To a cooled (0 °C) solution of (2-(8-oxa-3-azabicyclo[3.2.1]octan-3yl)pyridin-3-yl)methanol (0.070 g, 0.32 mmol) in dichloromethane was added SOCI₂ (0.23 mL, 3.2 mmol) and the reaction mixture was allowed to warm to ambient température. After 1 h, the reaction mixture was concentrated and azeotroped with toluene three times to provide 3-(3-(chloromethyl)pyridin-2-yl)-8-oxa-3-azabicyclo[3.2.1]octane (0.075 g, 98%) as a clear oil. MS (ES) for CnHxsCINîO: 239 (MH⁺).

[0136] Step 3: To a solution of provide 3-(3-(chloromethyl)pyridin-2-yl)-8-oxa-3azabicyclo[3.2.1]octane (0.080 g, 0.35 mmol) and 5-hydroxy-2,2-dimethyl-4H-

benzo[d][l,3]dioxin-4-one (0.061 g, 0.31 mmol) in DMF was added césium carbonate (0.307 g, .94 mmol) and the reaction mixture was heated (60 °C). After 30 minutes, the reaction mixture was partitioned between EtOAc and saturated aqueous sodium bicarbonate and the aqueous layer was extracted two times with EtOAc. Combined organic layers were washed with brine, dried over MGSO₄ and concentrated in vacuo. Purification by silica gel chromatography yielded 5-((2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)pyridin-3-yl)methoxy)2,2-dimethyl-4H-benzo[d][l,3]dioxin-4-one ( 112 mg, 90% yield). MS (ES) for C22H24N2O5: 397 (MH⁺).

[0137] Step 4:To a cooled (-78 °C) solution of 5-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8yl)pyridin-3-yl)methoxy)-2,2-dimethyl-4H-benzo[d][l,3]dioxin-4-one (0.11 g, 0.28 mmol) in CH2CI2 was added DIBAL-H (0.85 mL, IM in 0Η₂Ο₂) and reaction mixture was allowed to warm to ambient température over 3 hours. The reaction mixture was then cooled (-78 °C) and MeOH was added followed by saturated potassium sodium tartrate solution (300 pL). This mixture was stirred for 2 hours at ambient température and filtered over Celite. The resulting solution was partitioned between EtOAc and saturated aqueous NaHCO3 and washed two times with EtOAc. The combined organic layers were washed with brine, dried over MgSO₄ and concentrated in vacuo. Purification by preparatory HPLC resulted in 2-((2(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)pyridin-3-yl)methoxy)-6-hydroxybenzaldehyde (0.025 g, 25% yield). ^XH NMR (400 MHz, Chloroform-d) δ 11.95 (s, IH), 10.39 (d, J = 0.6 Hz, IH), 8.32 (dd, J = 4.8, 1.9 Hz, IH), 7.74 (dd, J = 8.0, 2.1 Hz, IH), 7.40 (t, J = 8.4 Hz, IH), 7.00 (dd, J = 7.5, 4.8 Hz, IH), 6.56 (d, J = 8.5 Hz, IH), 6.39 (d, J = 8.3 Hz, IH), 5.15 (s, 2H), 4.47 - 4.40 (m, 2H), 3.33 (dd, J = 12.5, 2.0 Hz, 2H), 3.03 (dd, J = 12.3,1.4 Hz, 2H), 2.13 - 1.94 (m, 4H). MS (ES) for Q9H20N2O4: 341 (MH⁺).

GBT911

GBT911

OH

[0138] GBT911-2-((2-(8-oxa-3-azabicydo[3.2.1]octan-3-yl)pyridin-3-yl)methoxy)-6hydroxybenzaldehyde. The compound was prepared from ethyl 2-fluoronicotinate and (S)3-fluoropyrrolidine according to réaction scheme below.

OH O

[0139] Step la: To a solution of ethyl 2-fluoronicotinate (0.090 g, 0.58 mmol) in DMF (0.3 mL) was added diisopropylethyl amine (0.51 mL, 2.9 mmol), and (S)-3-fluoropyrrolidine (0.10 g, 1.2 mmol). The resulting mixture was irradiated with microwaves (100 °C) for lh and loaded directly onto a silica column. Eluting the column with EtOAc/hexanes (0-100%) provided (S)-ethyl 2-(3-fluoropyrrolidin-l-yl)nicotinate as a clear oil (0.100 g, 46% yield). MS (ES) for Ci₂Hi5FN₂O₂: 225 (MH⁴).

[0140] Step lb: To a cooled (0 °C) solution of (S)-methyl 2-(3-fluoropyrrolidin-lyljnicotinate (0.20 g, 0.87 mmol) in THF (5 mL) was added a solution of lithium aluminum hydride (2.6 mL, IM in THF). The reaction mixture was stirred for lh and then 20 pL of H₂O was added followed by 20 pL of 15% NaOH (aq) and then 60 pL of additional H₂O. The slurry was stirred for lh, filtered and the resulting residue was washed with ether. The combined organic layers were dried over MgSO₄ and concentrated in vacuo. Purification by column chromotography (EtOAc/hexanes,0-100%) provided (S)-(2-(3-fluoropyrrolidin-l-yl)pyridin-3yljmethanol (0.165 g, 97% yield). MS (ES) for Ci₀H₁₃FN₂O: 197 (MH⁺).

[0141] Step 2: To a cooled (0 °C) solution of (S)-(2-(3-fluoropyrrolidin-l-yl)pyridin-3yl)methanol (0.081 g, 0.77 mmol) in dichloromethane was added SOCI₂ (0.92 g, 7.7 mmol) and the reaction mixture was allowed to warm to ambient température. After 1 h, the reaction mixture was concentrated and azeotroped with toluene to provide (S)-3-

(chloromethyl)-2-(3-fluoropyrrolidin-l-yl)pyridine (0.180 g, 99%) as a clear oil. MS (ES) for Ci_OHi₂CIFN₂: 215 (MH⁺).

[0142] Step 3: To a solution of provide (S)-3-(chloromethyl)-2-(3-fluoropyrrolidin-lyl)pyridine (0.085 g, 0.40 mmol) and 5-hydroxy-2,2-dimethyl-4H-benzo[d][l,3]dioxin-4-one (0.12 g, 0.59 mmol) in DMF was added césium carbonate (0.39 g, 0.12 mmol) and the reaction mixture was heated (60 °C). After 30 minutes, the reaction mixture was partitioned between EtOAc and saturated aqueous sodium bicarbonate and the aqueous layer was extracted two times with EtOAc. Combined organic layers were washed with brine, dried over MGSO4 and concentrated in vacuo. Purification by silica gel chromatography yielded (S)-5-((2-(3-fluoropyrrolidin-l-yl)pyridin-3-yl)methoxy)-2,2-dimethyl-4H-benzo[d][l,3]dioxin4-one (120 mg, 81% yield). MS (ES) for C20H21FN2O4: 373 (MH⁺).

[0143] Step 4: To a cooled (-78 °C) solution of (S)-5-((2-(3-fluoropyrrolidin-l-yl)pyridin-3yl)methoxy)-2,2-dimethyl-4H-benzo[d][l,3]dioxin-4-one (0.085 g, 0.23 mmol) in CH2CI2 was added DIBAL-H (0.68 mL, IM in CH2CI2) and reaction mixture was allowed to warm to ambient température over 3 hours. The reaction mixture was then cooled (-78 °C) and

MeOH was added followed by saturated potassium sodium tartrate solution (300 pL). This mixture was stirred for 2 hours at ambient température and filtered over Celite. The resulting solution was partitioned between EtOAc and saturated aqueous NaHCO3 and washed two times with EtOAc. The combined organic layers were washed with brine, dried 20 over MgSO₄ and concentrated in vacuo. Purification by preparatory HPLC resulted in 2-((2(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)pyridin-3-yl)methoxy)-6-hydroxybenzaldehyde (0.020 g, 28% yield). *H NMR (400 MHz, Chloroform-d) δ 11.97 (s, 1H), 10.34 (s, 1H), 8.21 (dd, J = 4.8, 1.9 Hz, 1H), 7.56 (ddd, J = 7.4, 1.9, 0.5 Hz, 1H), 7.42 (t, J = 8.4 Hz, 1H), 6.75 (dd, J = 7.4, 4.8 Hz, 1H), 6.57 (d, J = 8.0 Hz, 1H), 6.44 (d, J = 9.0 Hz, 1H), 5.24 (dt, J = 53.0, 3.9, 3.3 Hz, 1H),

5.16 (d, J = 11.4 Hz, 1H), 5.05 (d, J = 11.4 Hz, 1H), 3.97 - 3.60 (m, 4H), 2.37 -1.96 (m, 2H).

MS (ES) for C17H17FN2O3: 317 (MH⁺).

[0144] G BT1028 - 2-hyd roxy-5-((2^,,2^,,6',6^,-tetramethyl-l^,,2',3',6'-tetrahydro-[2,4'bipyridin]-3-yl)methoxy)benzaldehyde. The compound was prepared by Suzuki coupling of 2,2,6,6-tetramethyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,2,3,6tetrahydropyridine and 2-((2-bromopyridin-3-yl)methoxy)-65 (methoxymethoxy)benzaldehyde according to scheme 4, reaction step 2; the MOM ether protecting group was removed by treating with conc HCl (2eq) in THF. The product HCl sait was obtained as brown solid after silica gel chromatography. ³H NMR (400 MHz, DMSO-d₆) δ 11.70 (s, IH), 10.30 (s, IH), 9.21 (s, 2H), 8.62 (dd, J = 4.9, 1.6 Hz, IH), 8.24 - 8.16 (m, IH), 7.58 - 7.46 (m, 2H), 6.67 (d, J = 8.3 Hz, IH), 6.56 (d, J = 8.4 Hz, IH), 5.94 (d, J = 1.8 Hz, IH), 5.26 (s, 2H), 3.66-3.54 (m,2H), 1.56 - 1.37 (m, 12H); MS (ES, m/z) 367.38 [M+l]⁺.

[0145] GBT1045-2-hydroxy-6-((2-(4-methylpiperazin-l-yl)pyridin-3yl)methoxy)benzaldehyde. The compound was prepared from methyl 2-chloronicotinate and methylpiperazine according to scheme 5, reaction steps 1 and 2.

[0146] Step la: Into a 100-mL round-bottom flask, was placed a solution of methyl 2chloropyridine-3-carboxylate (2.0 g, 11.66 mmol, 1.00 equiv) in N,N-dimethylformamide (40 mL). 1-methylpiperazine (1.75 g, 17.47 mmol, 1.50 equiv), potassium carbonate (3.30 g,

23.88 mmol, 2.00 equiv), 18-crown-6 (200 mg, 0.06 equiv) were added to the reaction. The resulting solution was stirred overnight at 100°C. The reaction mixture was cooled to room température. The resulting solution was diluted with 30 mL of H₂O, and then it was extracted with 5x30 mL of ethyl acetate. The combined organic layers were concentrated . under vacuum. The residue was applied onto a silica gel column with dichloromethane/methanol (10:1) as eluent. This resulted in 2.7 g (98%) of methyl 2-(4methylpiperazin-l-yl)pyridine-3-carboxylate as a yellow oil.

[0147] Step lb: Into a 100-mL round-bottom flask, was placed a solution of methyl 2-(4methylpiperazin-l-yl)pyridine-3-carboxylate (1.3 g, 5.53 mmol, 1.00 equiv) in θ tetrahydrofuran (40 mL). This was followed by the addition of Al Li H₄ (315 mg, 8.30 mmol,

1.50 equiv) at 0°C. The resulting solution was stirred for 5 h at 0°C, and then it was quenched by the addition of 0.5 mLof water, 1.5 ml of NaOH(15%) and 0.5 ml of water. The solids were fiîtered out. The resulting mixture was concentrated under vacuum. The residue was applied onto a silica gel column with dichloromethane/methanol (1:1) as eluent. This resulted in 500 mg (44%) of [2-(4-methylpiperazin-l-yl)pyridin-3-yl]methanol as a yellow solid.

[0148] Step 2: Into a 50-mL round-bottom flask, was placed a solution of [2-(4methylpiperazin-l-yl)pyridin-3-yl]methanol (200 mg, 0.96 mmol, 1.00 equiv) in tetrahydrofuran (20 mL). 2,6-Dihydroxybenzaldehyde (200 mg, 1.45 mmol, 1.50 equiv) and 2q PPh₃ (380 mg, 1.45 mmol, 1.50 equiv) were added to the réaction. This was followed by the addition of DIAD (293 mg, 1.45 mmol, 1.50 equiv) at 0°C. The resulting solution was stirred overnight at room température, and then it was concentrated under vacuum. The crude product (200 mg) was purified by Prep-HPLC with the following conditions (Prep-HPLC-010): Column, SunFire Prep C18 OBD Column, 5um, 19*150mm,; mobile phase, water with

0.05%TFA and MeCN (25.0% MeCN up to 42.0% in 13 min, up to 95.0% in 2 min, down to

25.0% in 2 min); Detector, Waters2545 UvDector 254&220nm. This resulted in 67.9 mg (21%) of 2-hydroxy-6-[[2-(4-methylpiperazin-l-yl)pyridin-3-yl]methoxy]benzaldehyde as a yellow oil; ¹HNMR (400MHz, CDCl₃, ppm): 11.98 (s, 1H), 10.43 (s, 6H), 8.35(m, 1H), 7.77(d, J=5.7Hz, 1H), 7.42(m, 1H), 7.03 (m, 1H), 6.58(d,7=6.3Hz, 1H), 6.43(d, J=6.0Hz, 1H), 5.18(d, _3Q J=7.8Hz, 2H), 3.26 (m, 4H), 2.64 (s, 4H), 2.40 (s, 3H) 1.42-2.09(m, 8H); MS (ES, m/z): 328 [M+l]⁺.

[0149] GBT1249- 2-((2-chloropyridin-3-yl)methoxy)-6-(methoxymethoxy)benzaldehyde.

The compound was prepared by O-alkylation of 2-hydroxy-6(methoxymethoxy)benzaldehyde and 2-chloro-3-(chloromethyl)pyridine. The product as white solid was obtained after flash column purification. ’HNMR (400MHz, CDCI₃, ppm): 10.65(s, 1H), 8.37(d, J=5.7Hz, 1H), 7.49(t, J=6.3Hz, 1H), 7.39(t, J=4.5Hz, 1H), 7.28(s, 1H), 6.90(d, J=6.3Hz, 1H), 6.75(d, J=6.3Hz, 1H), 5.32(s, 2H), 5.21(s, 2H), 3.54(s, 3H); MS (ES, m/z): 308[M+l]⁺

OMOM

GBT001046

[0150]

GBT1046- 2-((2-(3,6-dihydro-2H-pyran-4-yl)pyridin-3-yl)methoxy)-6hydroxybenzaldehyde.

1) Suzuki

ΌΜΟΜ

ΌΗ

[0151] The compound was prepared by Suzuki coupling of 2-(3,6-dihydro-2H-pyran-4-yl)4,4,5,5-tetramethyl-l,3,2-dioxaborolane and 2-((2-bromopyridin-3-yl)methoxy)-6(methoxymethoxy)benzaldehyde according to scheme 4, reaction step 2; the MOM ether protecting group was removed by treating with conc HCl (2eq) in THF. The product was obtained as light brown solid after silica gel chromatography. *H NMR (400 MHz, Chloroform-d) d 11.93 (d, J = 0.6 Hz, 1H), 10.37 (s, 1H), 8.84 (s, 1H), 8.56 (d, J = 7.2 Hz, 1H), 7.89 (s, 1H), 7.46 (t, J = 8.3 Hz, 1H), 6.67 (d, J = 8.5 Hz, 1H), 6.36 (d, J = 7.6 Hz, 2H), 5.29 (s, 2H), 4.43 (s, 2H), 4.08 (t, J = 4.5 Hz, 2H), 2.80 (s, 2H); MS (ES, m/z) 312.33 [M+lf.

[0152] GBT1063- 2-hydroxy-6-((2-(4-methyl-l,4-diazepan-l-yl)pyridin-3yl)methoxy)benzaldehyde. The compound was prepared from methyl 2-chloronicotinate and l-methyl-l,4-diazepane according to scheme 5, reaction steps 1 and 2.

[0153] Step la: Into a 100-mL round-bottom flask, was placed a solution of methyl 2chloropyridine-3-carboxylate (2.0 g, 11.66 mmol, 1.00 equiv) in N,N-dimethylformamide (40 mL). l-methyl-l,4-diazepane (2.0 g, 17.51 mmol, 1.50 equiv), potassium carbonate (3.3 g, 23.88 mmol, 2.00 equiv), and 18-crown-6 (200 mg, 0.06 equiv) were added to the reaction. The resulting solution was stirred ovemight at 100°C. The réaction mixture was cooled to room température, and then it was diluted with 40 mL of H₂O. The resulting solution was extracted with 5x30 mL of ethyl acetate and the combined organic layers were concentrated under vacuum. The residue was applied onto a silica gel column with

dichloromethane/methanol (10:1) as eluent. This resulted in 2.65 g (91%) of methyl 2-(4methyl-l,4-diazepan-l-yl)pyridine-3-carboxylate as a yellow oil.

[0154] Step lb: Into a 100-mL round-bottom flask, was placed a solution of methyl 2-(4methyl-l,4-diazepan-l-yl)pyridine-3-carboxylate (1.2 g, 4.81 mmol, 1.00 equiv) in tetrahydrofuran (40 mL). This was followed by the addition of LiAIH₄ (500 mg, 13.18 mmol, 2.00 equiv) at 0°C. The resulting solution was stirred for 2 h at room température. The reaction was then quenched by the addition of 0.5 mL of water, 1.5 mL of 15% NaOH,0.5 mL of H₂O. The solids were filtered out. The resulting mixture was concentrated under vacuum. The residue was applied onto a silica gel column with dichloromethane/methanol (3:1) as eluent. This resulted in 800 mg (75%) of [2-(4-methyl-l,4-diazepan-l-yl)pyridin-3yljmethanol as a yellow oil.

[0155] Step 2: Into a 50-mL round-bottom flask, was placed a solution of [2-(4-methyl-l,4diazepan-l-yl)pyridin-3-yl]methanol (300 mg, 1.36 mmol, 1.00 equiv) in tetrahydrofuran (25 mL). 2,6-Dihydroxybenzaldehyde (280 mg, 2.03 mmol, 1.50 equiv) and PPhî (532 mg, 2.03 mmol, 1.50 equiv) were added to the reaction. This was followed by the addition of DIAD (410 mg, 2.03 mmol, 1.50 equiv) at 0°C. The resulting solution was stirred overnight at room température, and then it was concentrated under vacuum. The crude product (300 mg) was purified by Prep-HPLC with the following conditions (Prep-HPLC-010): Column, Gemini-NX 150*21.20mm C18 AXIA Packed, 5um 110A; mobile phase, water with O.O596TFA and MeCN (10.0% MeCN up to 50.0% in 5 min); Detector, nm. This resulted in 159.5 mg (34%) of 2hydroxy-6-[[2-(4-methyl-l,4-diazepan-l-yl)pyridin-3-yl]methoxy]benzaldehyde as a yellow oil; ¹HNMR (400MHz, DMSO+D₂O, ppm): 10.29 (s, IH), 8.19 (d, J=2.7Hz, IH), 7.95 (d, J=5.4Hz, IH), 7.52 (m, lH),7.08(m, IH), 6.66 (d, J=6.3Hz, IH), 6.57 (d, J=0.9Hz, IH), 5.21(s, 2H), 3.74(s, 2H), 3.45(m, 6H), 2.84(s, 3H), 2.11 (d, J=3.9Hz, 2H); (ES, m/z ): 342 [M+l]⁺.

GBT001121

ΌΗ

[0156] GBT1121-2-((2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)pyridin-3-yl)methoxy)-6hydroxybenzaldehyde. The compound was prepared from methyl 2-fluoronicotinate and 2oxa-6-azaspiro[3.3]heptane according to scheme 5, reaction steps 1 and 2.

[0157] Step la: Methyl 2-fluoronicotinate (0.3 g, 1.93 mmol) and 2-oxa-65 azaspiro[3.3]heptane oxalate (0.55 g, 2.9 mmol) were combined with DMF (3 ml). N,Ndiisopropylethylamine (2 ml, 11.6 mmol) was added and the mixture was heated in a microwave reactor (120 °C, 1 h). Ethyl acetate (100 ml) and water (50 ml) were added to the cooled solution and the phases were separated. The aqueous phase was extracted with ethyl acetate (2 x 50 ml). The combined organic phases were washed with water (30 ml) and a saturated aqueous sodium chloride solution (30 ml), and dried over sodium sulfate. After évaporation, the residue was purified by silica gel chromatography (5 - 80% ethyl acetate/hexanes) to give 0.27 g (59%) of methyl 2-(2-oxa-6-azaspiro[3.3]heptan-6yl)nicotinate as a white solid. MS (ESI) m/z 235 [M+H]⁺.

[0158] Step lb: Methyl 2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)nicotinate (0.26 g, 1.1 mmol)

-15 was dîssolved in THF (5 ml) and stirred in an ice bath. Lithium aluminum hydride (2.2 ml of a

IM THF solution) was added dropwise. The reaction was stirred to 25 °C over 2 h. Water (0.084 ml) was carefully added followed by 15% aqueous sodium hydroxide solution (0.084 ml) and water (0.25 ml). The mixture was stirred for 30 m then filtered, rinsed with THF (10 ml) and the solvent evaporated to give 226 mg (98%) of (2-(2-oxa-6-azaspiro[3.3]heptan-620 yl)pyridin-3-yl)methanol which was used directly in the next step. MS (ESI) m/z 207 [M+H]⁺.

[0159] Step 2: 2-(2-Oxa-6-azaspiro[3.3]heptan-6-yl)pyridin-3-yl)methanol (0.12 g, 0.582 mmol) 2,6-dihydroxybenzaldehyde (96 mg, 0.7 mmol) and triphenylphosphine-polystyrene resin (0.63 g, 0.76 mmol) were combined with THF (3 ml), and stirred in an ice bath. Diisopropylazodicarboxylate (0.15 ml, 0.76 mmol) was added dropwise and the reaction was stirred to 25 °C over 16 h. The reaction was filtered, rinsed with THF (10 ml) and evaporated.

The resulting residue was purified by silica gel chromatography (0 -75 % ethyl acetate/dichloromethane) to give 31 mg (16%) of 2-((2-(2-oxa-6-azaspiro[3.3]heptan-6-

yl)pyridin-3-yl)methoxy)-6-hydroxybenzaldehyde as a white solid after lyophilization from acetonitrile/water. ^XH NMR (400 MHz, CDCI₃) δ 11.97 (s, 1H), 10.36 (s, 1H), 8.21 (dd, J= 1.65,

4.92 Hz, 1H), 7.51 (dd, J = 1.68, 7.37 Hz, 1H), 7.44 (t, J = 8.38 Hz, 1H), 6.76 (dd, J = 4.95, 7.34

Hz, 1H), 6.60 (d, J = 8.49 Hz, 1H), 6.42 (d, J = 8.28 Hz, 1H), 4.96 (s, 2H), 4.81 (s, 4H), 4.27 (s,

4H). MS (ESI) m/z 327 [M+H]⁺.

[0160] GBT1122- 2-Hydroxy-6-((2-morpholinopyridin-3-yl)methoxy)benzaldehyde. The compound was prepared from ethyl 2-fluoronicotinate and morpholine according to a modified scheme 5, reaction steps 1, 3 and 4.

[0161] Step la: To a solution of ethyl 2-fluoronicotinate (0.40 g, 2.6 mmol) in DMF (0.3 mL) was added diisopropylethyl amine (1.8 mL, 10 mmol), and morpholine (0.45 g, 5.2 mmol). The resulting mixture was irradiated with microwaves (100 °C) for lh and loaded directly onto a silica column. Eluting the column with EtOAc/hexanes (0-100%) Methyl 2morpholinonicotinate as a clear oil (0.36 g, 62% yield). MS (ES) for C12H16N2O3: 237 (MH⁺). [0162] Step lb: To a cooled (0 °C) solution of Methyl 2-morpholinonicotinate (0.36 g, 1.6 mmol) in THF (5 mL) was added a solution of lithium aluminum hydride (4.9 mL, IM in THF). The reaction mixture was stirred for lh and then 180 pL of H2O was added followed by 180 pL of 15% NaOH (aq) and then 540 pL of additional water. The slurry was stirred for lh and filtered and the resulting residue was washed with ether. The combined organic layers were dried over MgSO₄ and concentrated in vacuo. Purification by column chromotography (EtOAc/hexanes,0-100%) provided (2-morpholinopyridin-3-yl)methanol (0.224 g, 71% yield). MS (ES) for CioHi₄N₂0₂: 195 (MH⁺).

[0163] Step 3: To a cooled (0 °C) solution of provided (2-morpholinopyridin-3-yl)methanol (0.100 g, 0.51 mmol) in dichloromethane was added SOCI₂ (0.50 mL, 6.9 mmol) and the reaction mixture was allowed to warm to ambient température. After 1 h, the reaction mixture was concentrated and azeotroped with toluene to provide 4-(3(chloromethyl)pyridin-2-yl)morpholine (0.11 g, 96%) as a clear oil. MS (ES) for Ci₀Hi₃CIN₂O: 213 (MH⁺).

[0164] Step 4: To a solution of 4-(3-(chloromethyl)pyridin-2-yl)morpholine (0.11 g, 0.50 mmol) and 2-hydroxy-6-(methoxymethoxy)benzaldehyde (0.09 g, 0.50 mmol) in DMF was added potassium carbonate (0.210 g, 1.5 mmol) and the reaction mixture was heated (60 °C). After 30 minutes, the reaction mixture was partitioned between EtOAc and saturated NaHCOî and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with brine, dried over MgSO₄ and concentrated in vacuo to yield 2(methoxymethoxy)-6-((2-morpholinopyridin-3-yl)methoxy)benzaldehyde ( 0.145 mg, 80% yield) as a white powder. MS (ES) for CigH₂₂N₂O₅: 359 (MH⁺).

[0165] Step 5: To a solution of 2-(methoxymethoxy)-6-((2-morpholinopyridin-3yl)methoxy)benzaldehyde (0.120 g, 0.33 mmol) in THF (5 mL) was added concentrated HCl (0.5 mL, 6 mmol). After stirring at ambient température for 3 hours, the mixture was partitioned between EtOAc and saturated aqueous NaHCO₃ and the aqueous phase was extracted twice with EtOAc. The combined organic layers were washed with brine, dried over MgSO₄and concentrated in vacuo. Purification, reaction silica gel chromatography provided 2-hydroxy-6-((2-morpholinopyridin-3-yl)methoxy)benzaldehyde (0.074 g, 0.24 mmol) as a white powder. Y NMR (400 MHz, Chloroform-c/) δ 11.95 (s, 1H), 10.40 (s, 1H), 8.34 (dd, J = 4.8,1.9 Hz, 1H), 7.77 (dd, J = 7.5,1.7 Hz, 1H), 7.40 (t, J = 8.4 Hz, 1H), 7.04 (dd, J = 7.5, 4.9 Hz, 1H), 6.56 (d, J = 8.5 Hz, 1H), 6.40 (d, J = 8.3 Hz, 1H), 5.15 (s, 2H), 3.90 - 3.83 (m, 3H), 3.22 - 3.15 (m, 4H). MS (ES) for C₁₇H₁₈N₂O₄: 315 (MH⁺).

[0166] From the foregoing it will be appreciated that, although spécifie embodiments of the invention hâve been described herein for purposes of illustration, various modifications may be made without deviating from the spirit and scope of the invention.

[0167] Throughout the description of this invention, reference is made to various patent applications and publications, each of which are herein incorporated by reference in their entirety.

Claims (4)

¹ □ As ail required addtional search fees were timely paid by the applicant, this international search report covers ail searchable claims.

As ail searchable claims could be searched without effort justifying an additional fees, this Authority did not invite payment of any additional fees.

1. β/Ι Claims Nos.: 10, 11 because they relate to subject matter not required to be searched by this Authority, namely:

Claims 10 and 11 pertain to a method for treatment of the human body by therapy and thus relate to a subject matter which this International Searching Authority is not required, under PCT Article 17(2)(a)(i) and PCT Rule 39.1(iv), to search.

1-5,7,8 | | Further documents are listed in the continuation of Box C.

* Spécial categories of cited documents:

A document defining the general state of the art which is not considered to be of particular relevance

E eariier application or patent but published on or after the international filing date

L document which may throw doubts on priority claim(s) or which is cited to establish the publication date of another citation or other spécial reason (as specified)

O document referring to an oral disclosure, use, exhibition or other means

P” document published prior to the international filing date but later than the priority date claimed

See patent family annex.

T later document published after the international filing date or priority date and not in conflict with the application but cited to understand the principle or theory underiying the invention

X document of particular relevance; the claimed invention cannot be considered novel or cannot be considered to involve an inventive step when the document is taken alone

Y document of particular relevance; the claimed invention cannot be considered to invotve an inventive step when the document is combined with one or more other such documents,such combination being obvious to a person skilled in the art & document member of the same patent family

Date of the actual completion of the international search

19 August 2014 (19.08.2014)

Date of mailing of the international search report

19 August 2014 (19.08.2014)

Name and mailing address of the ISA/KR International Application Division Korean Intellectual Property Office 189 Cheongsa-ro, Seo-gu, Daejeon Metropolitan City, 302-701, Republic of Korea

FacsimileNo. +82-42-472-7140

Àuthorized officer

CHOI, Sung Hee

Téléphoné No. +82-42-481-8740

FormPCT/ISA/210 (second sheet) (July 2009)

International application No.

PCT/US2014/022736

INTERNATIONAL SEARCH REPORT

Box No. II Observations where certain claims were found unsearchable (Continuation of item 2 of first sheet)

This international search report has not been established in respect of certain claims under Article 17(2)(a) for the following reasons:

1-5,7,8

1-5,7,8

1-5,7,8

1-5,7,8

1-5,7,8

1. In certain aspects of the invention, a compound of formula (I) is provided:

or a tautomer thereof, or a pharmaceutically acceptable sait of each thereof, wherein ring A is an optionally substituted 4-10 membered cycloalkyl or 4-10 membered heterocycle containing up to 5 ring heteroatoms, wherein the heteroatom is selected from the group consisting of O, N, S, and oxidized forms of N and S;

ring B is a C6-Ci_Oaryl or 5-10 membered heteroaryl having 1-3 nitrogen atoms, preferably 1-2 nitrogen atoms and more preferably 1 nitrogen atom, or oxidized versions thereof, wherein the aryl or heteroaryl is optionally substituted;

each Y and Z is independently CR¹⁰Rⁿ,0, S, SO, SO2, or NR¹²; each R¹⁰ and R¹¹ independently is hydrogen or C1-C3 alkyl optionally substituted with halo, OH, or Ci-C₆ alkoxy, or CR^1OR^U is C=O; R¹² is hydrogen or Ci-Ce alkyl; provided that if one of Y and Z is O, S, SO, SO2, then the other is not CO, and provided that Y and Z are both not heteroatoms or oxidized forms thereof;

ring C is C₆-Ci₀ aryl, optionally substituted;

V¹ and V² independently are Ci-C₆ alkoxy; or V¹ and V² together with the carbon atom they are attached to form a ring of formula:

wherein each V³ and V⁴are independently O, S, or NH, provided that when one of V³ and V⁴ is S, the other is NH, and provided that V³ and V⁴ are both not NH; q is 1 or 2; each V⁵ is independently Ci-Cg alkyl or CO2R⁶⁰, where each R⁶⁰ independently is C1-C6 alkyl or hydrogen; t is 0,1, 2, or 4; or CV^XV² is C=V, wherein V is O, NOR⁸⁰, or NNR⁸¹R⁸²;

R⁸⁰ is optionally substituted Ci-Ce alkyl;

R⁸¹ and R⁸² independently are selected from the group consisting of hydrogen, optionally substituted C1-C6 alkyl, COR⁸³, or CO2R⁸⁴;

R⁸³ is hydrogen or optionally substituted C_rC₆ alkyl; and

R⁸⁴ is optionally substituted Ci-Cg alkyl.

2. I I Claims Nos.:

— because they relate to parts of the international application that do not comply with the prescribed requirements to such an extent that no meaningful international search can be carried out, specifically:

2. The compound of claim 1, wherein V¹ and V² independently are Ci-Q alkoxy; or V¹ and V² together with the carbon atom they are attached to form a ring of formula: wherein each V³ and V⁴ are independently O, S, or N H, provided that when one of V³ and V⁴ is S the other is NH, and provided that V³ and V⁴ are both not NH; q is 1 or 2; each V⁵ is independently Ci-C₆ alkyl or CO₂R⁶⁰, where each R⁶⁰ independently is Ci-Ce alkyl or hydrogen; t is 0,1, 2, or 4; or CV¹)/² is C=V, wherein V is O, and wherein the remaining variables are defined as in claim 1.

3· Π As only some of the required additional search fees were timely paid by the applicant, this international search report covers only those claims for which fees were paid, specifically claims Nos.:

3. [><] Claims Nos.: 6, 9-11 because they are dépendent claims and are not drafied in accordance with the second and third sentences of Rule 6.4(a).

Box No. ΙΠ Observations where unity of invention is lacking (Continuation of item 3 of first sheet)

This International Searching Authority found multiple inventions in this international application, as follows:

3.

The compound of claim 2 of formula (11):

wherein R⁵ is hydrogen, Ci-C₆ alkyl or a prodrug moiety R, wherein the Ci-C₆ alkyl is optionally substituted with 1-5 halo;

R⁶ is halo, CpCe alkyl, Ci-Cg alkoxy, Ci-Cg alkylthio, Ci-C₆ S(O)-, Ci-Ce S(O)2-,wherein the Ci-C₆ alkyl is optionally substituted with 1-5 halo; or

R⁶ is 4-10 membered cycloalkyl or heterocycle substituted with an R'R'N- moiety

5 wherein each R' is independently Q-Cg alkyl or hydrogen;

p is 0,1, 2 or 3; and the remaining variables are defined as in claim 2.

4. A compound of claim 2 or 3 of Formula (IIA):

(II) wherein the variables are defined as in claims 2 and 3.

5. The compound of claim 2, wherein ring A is optionally substituted with 1-3: halo, C_r Ce alkyl, COR¹⁵ and/or COOR¹⁵; wherein R¹⁵ is optionally substituted Ci-C₆ alkyl, optionally substituted Cg-Cioaryl, optionally substituted 5-10 membered heteroaryl containing up to 5 5 ring heteroatoms, or optionally substituted 4-10 membered heterocycle containing up to 5 ring heteroatoms, wherein the heteroatom is selected from the group consisting of O, N, S, and oxidized forms of N and S.

6. The compound of claim 4 or 5, wherein ring B is optinally substituted with 1-3: halo, Ci-C₆ alkyl COR¹⁵ and/or COOR¹⁵; wherein R¹⁵ is optionally substituted CpCg alkyl, optionally

10 substituted C6-Ci₀aryl, optionally substituted 5-10 membered heteroaryl containing up to 5 ring heteroatoms, or optionally substituted 4-10 membered heterocycle containing up to 5 ring heteroatoms, wherein the heteroatom is selected from the group consisting of O, N, S, and oxidized forms of N and S.

7. The compound of claim 2, wherein the compound is selected from the group consisting of or an N oxide thereof, wherein

R¹⁴ is Ci-C₆ alkyl, C₃-C₈ cycloalkyl, COR¹⁵ or COOR¹⁵;

R¹⁵ is optionally substituted Ci-Ce alkyl, optionally substituted C₆-Cioaryl, optionally substituted 5-10 membered heteroaryl containing up to 5 ring heteroatoms, or

20 optionally substituted 4-10 membered heterocycle containing up to 5 ring heteroatoms, wherein the heteroatom is selected from the group consisting of O, N, S, and oxidized forms of N and S;

x is 0,1, or 2;

p is 0,1, and 2; and m is 0,1 or 2.

8. A compound of claim 1 selected from the group consisting of:

or a prodrug thereof, or a pharmaceuticlaly acceptable sait of each thereof.

9. A composition comprising a compound of any one of claims 2-8 and at least one pharmaceutically acceptable excipient.

5 10. Use of a compound of any one of claims 2 - 8 in the manufacture of a composition for încreasing oxygen affinity of hemoglobin S in a subject.

H, Use of a compound of any one of claims 2 - 8 in the manufacture of a composition for treating oxygen deficiency associated with sickle cell anémia.

International application No.

PCI7US2014/022736

CLASSIFICATION OF SUBJECT MATTER ^FA61K 31/444(2006.01)1, A61K 31/4427(2006.01)i, A61K 31/497(2006.01)i, A61P 3/00(2006.01)1, A61P 7/00(2006.01)1

INTERNATIONAL SEARCH REPORT

According to International Patent Classification (IPC) or to both national classification and IPC

B. FIELDS SEARCHED

Minimum documentation searched (classification system followed by classification symbols)

A61K 31/444; C07C 233/00; C07D 487/04; A61K 31/58; A61K 31/55; A61K 31/11; A01N 35/00; C07D 495/04; A61K 48/00; A61K 31/437; C07C 233/01; A61K 31/5377; A61K 31/4427; A61K 31/497; A61P 3/00; A61P 7/00

Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched Korean utility models and applications for utility models Japanese utility models and applications for utility models

Electronic data base consulted during the international search (name of data base and, where practicable, search terms used) eKOMPASS(KIPO internai) & Keywords: hemoglobin, allosteric modulator, sickle cell disease, benzaldehyde

C. DOCUMENTS CONSIDERED TO BE RELEVANT

Categoiy*

Citation of document, with indication, where appropriate, of the relevant passages

US 2003-0060425 Al (AHLEM, C. N. et al.) 27 March 2003 See paragraph [0590],

US 2012-0220569 Al (OHASHI, T. et al.) 30 August 2012 See abstract! claims 1-8; and example 128.

US 2007-0293698 Al (QUICK, A. et al.) 20 December 2007 See abstract; claim 1; and figures 1A-1E.

WO 2012-138981 A2 (TEVA PHARMACEUTICAL INDUSTRIES LTD.) 11 October 2012 See abstract and claims 6-12.

US 2009-0023709 Al (GILLESPIE, P. et al.) 22 January 2009 See abstract and claims 1-13.

PX

WO 2013-102142 Al (GLOBAL BLOOD THERAPEUTICS, INC. et al.) 04 July 2013 See paragraphs [0200] and [0201],

Relevant to claim No.

4- □ No required additional search fees were timely paid by the applicant. Consequently, this international search report is restricted to the invention first mentioned in the claims; it is covered by claims Nos. :

Remark on Protest

The additional search fees were accompanied by the applicant's protest and, where applicable, the payment of a protest fee.

The additional search fees were accompanied by the applicant's protest but the applicable protest fee was not paid within the time limit specified in the invitation.

No protest accompanied the payment of additional search fees.

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US 7847120 B2 WO 2005-102305 A2 WO 2005-102305 A3 WO 2005-102308 A2 WO 2005-102308 A3 07/12/2010 03/11/2005 10/08/2006 03/11/2005 12/01/2006 WO 2012-138981 A2 11/10/2012 CA 2832570 Al 11/10/2012 EP 2694514 A2 12/02/2014 US 2014-0094604 Al 03/04/2014 US 2014-027234 Al 30/01/2014 US 2009-0023709 Al 22/01/2009 AR 067538 Al 14/10/2009 AU 2008-277783 Al 22/01/2009 AU 2008-277783 B2 20/09/2012 CA 2693457 Al 22/01/2009 CN 101743226 A 16/06/2010 CN 101743226 B 10/10/2012 EP 2178845 A2 28/04/2010 EP 2178845 B1 19/06/2013 JP 2010-533670 A 28/10/2010 JP 5189165 B2 24/04/2013 KR 10-1158191 B1 19/06/2012 KR 10-2010-0034763 A 01/04/2010 PE 08132009 Al 27/06/2009 RU 2010-105299 A 27/08/2011 TW 2009-13992 A 01/04/2009 US 7790711 B2 07/09/2010 WO 2009-010416 A2 22/01/2009 WO 2009-010416 A3 05/03/2009 ZA 2010-00318 A 27/10/2010 WO 2013-102142 Al 04/07/2013 US 2013-190315 Al 25/07/2013

Form PCT/ISA/210 (patent family annex) (Juty 2009)