Antiviral heterocyclic compounds

Abstract

The present invention discloses compounds of formula (I) and pharmaceutically acceptable salts, esters or prodrugs thereof: # imgabs0, which are useful as inhibitors of human respiratory syncytial virus (HRSV) or human metapneumovirus (HMPV). The invention also relates to a pharmaceutical composition containing the compound, and the pharmaceutical composition is used for being administered to a subject suffering from HRSV or HMPV infection. The invention also relates to methods of treating HRSV or HMPV infection in a subject by administering a pharmaceutical composition comprising a compound of the invention.

Description

[0001] Related Applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 328,400, filed on April 7, 2022. The entire teachings of the above application are incorporated herein by reference. Technical Field

[0003] The present invention generally relates to compounds and pharmaceutical compositions useful as inhibitors of respiratory syncytial virus (RSV) and human metapneumovirus (HMPV). Background Art

[0004] Human respiratory syncytial virus (HRSV) is a negative-sense virus containing a non-segmented, single-stranded, linear RNA genome. As a paramyxovirus of two serotypes in the family Pneumoviridae, HRSV contains 10 genes encoding 11 proteins. The nucleocapsid protein (N), RNA polymerase protein (L), phosphoprotein (P), and transcription antitermination factor (M2-1) along with the RNA genome form a ribonucleoprotein (RNP) complex. Several small molecule compounds have been shown to target the RNP complex. In addition, the fusion protein (F), which is essential for viral attachment to the host, has been extensively studied. High-resolution structures of the F protein interacting with inhibitors have been obtained, while structural studies of the N protein are still in the early stages. As a direct result of HRSV protein studies, the F, L, and N proteins have been the main focus of drug discovery efforts.

[0005] The increase in HRSV drug discovery is due to HRSV being the leading cause of acute lower respiratory tract infections (ALRI) in patients of all ages. In addition to respiratory infections, patient groups at high risk during HRSV infection include the elderly, immunocompromised individuals, children under two years of age, and patients with chronic obstructive pulmonary disease (COPD) or chronic heart failure (CHF). Over a four-year period, HRSV was found to cause 177,500 hospitalizations and 14,000 deaths in the elderly U.S. population. It is well known that nearly all children will be infected with HRSV within the first three years of life, and HRSV infection is more severe in premature infants. In fact, HRSV is the most common cause of bronchiolitis and pneumonia in infants under one year of age in the United States. It is estimated that approximately 3.2 million children under the age of five are hospitalized and 66,000 die worldwide due to HRSV. HRSV is more associated with infant mortality and infant hospitalization under one year of age than influenza.

[0006] HRSV infection can also affect healthy individuals, and repeated HRSV infections can occur even within two months. Symptoms are similar to those of a cold in healthy people, but more severe cases can include fever, wheezing, shortness of breath and difficulty breathing, and cyanosis.

[0007] Currently, treatment options for HRSV infection are very limited, and there is no vaccine due to unsuccessful attempts to date. Palivizumab is a monoclonal antibody that has been approved for prevention, but its use is limited due to its high price. Palivizumab is generally used only in high-risk infants, such as premature infants or those with cardiopulmonary disease, but is only 60% effective in reducing hospitalizations. Ribavirin is approved as an inhaled treatment option, but has limited effectiveness and associated safety issues. Given the treatment options and the continued seasonality of HRSV epidemics, the development of new therapeutic agents to treat HRSV is desirable.

[0008] Several RSV fusion inhibitors are disclosed in the following publications: WO2010103306, WO2012068622, WO2013096681, WO2014060411, WO2013186995, WO2013186334, WO2013186332, WO2012080451, WO2012080450, WO201208 0449, WO2012080447, WO2012080446, WO2015110446, WO2017009316, J. Med. Chem. 2015, 58, 1630-1643, Bioorg. Med. Chem. Lett., 2015, 25, 976-981 and Nat. Commun., 2017, 8, 167. Examples of other N protein inhibitors for treating HRSV have been disclosed in the following publications: WO2004026843, J. Med. Chem. 2006, 49, 2311-2319 and J. Med. Chem. 2007, 50, 1685-1692. Examples of HRSV L protein inhibitors are disclosed in WO2011005842, WO2005042530, Antiviral Res. 2005, 65, 125-131, and Bioorg. Med. Chem. Lett. 2013, 23, 6789-6793. Examples of nucleoside / polymerase inhibitors are disclosed in WO2011005842, WO2013242525, WO2014031784, WO2015026792, WO20160055791, WO2016138158, and J. Med. Chem. 2015, 58, 1862-1878.

[0009] Similarly, human metapneumovirus (HMPV), discovered by van Den Hoogen in 2001, is a negative-sense, single-stranded RNA enveloped virus belonging to the Pneumoviridae family and the genus Metapneumovirus, and is also a common cause of acute lower respiratory tract infections (ALRTI). Although usually mild, this virus can be severe and life-threatening in high-risk groups, such as children under 5 years of age, elderly people over 65 years of age, and adults with underlying medical conditions such as chronic obstructive pulmonary disease (COPD), asthma, congestive heart failure, or diabetes. In healthy adults over 65 years of age, the annual incidence of HMPV infection is 1.2 / 1000, the annual incidence of disease such as COPD is 38%, and the annual incidence is twice that of individuals who may develop symptomatic disease and require medical care. In immunocompromised individuals, HMPV accounts for 6% of total respiratory infections in lung transplants and 3% of respiratory infections associated with stem cell transplants. HMPV infection has also been implicated in acute graft rejection.

[0010] As with HRSV, infection is thought to occur via attachment to target cells via glycoprotein (G) protein interactions followed by fusion via the F protein. The HMPV L protein sequence is homologous to the HRSV L protein.

[0011] HMPV infection is the second most common cause of lower respiratory tract infection in children (after HRSV) and is also a problem in the elderly. Four HMPV subtypes (A1, A2, B1, and B2) have been found in clinical isolates. After the initial infection, reinfections occur throughout childhood. There are currently no drugs to treat HMPV infection.

[0012] Considering the seasonality and predictability of HRSV and HMPV epidemics, the HRSV epidemics in elderly institutions, and the severity of infection in high-risk infants, it is clear that there is a need for strong and effective treatment of HRSV and HMPV. The present invention has identified heterocyclic molecule compounds that are potent against HRSV-A / B and HMPV. The present invention includes methods for preparing these molecules, RSV cell-based assays, HMPV-TN / 94-49A2 cell-based assays, and small molecules that may treat HRSV / HMPV infections. Summary of the invention

[0013] The present invention provides compounds represented by formula (I) and pharmaceutically acceptable salts, esters and prodrugs thereof that can be used to treat or prevent viral (particularly HRSV or HMPV) infection:

[0014]

[0015] or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein:

[0016] A is selected from:

[0017] 1) optionally substituted aryl; and

[0018] 2) optionally substituted heteroaryl;

[0019] E is selected from:

[0020] 1) optionally substituted aryl; and

[0021] 2) optionally substituted heteroaryl;

[0022] R1 and R2 are each independently selected from:

[0023] 1) Hydrogen;

[0024] 2) optionally substituted -C1-C8 alkyl;

[0025] 3) optionally substituted -C3-C8 cycloalkyl;

[0026] 4) optionally substituted 3 to 8 membered heterocycloalkyl;

[0027] 5) optionally substituted aryl;

[0028] 6) optionally substituted aralkyl;

[0029] 7) optionally substituted heteroaryl; and

[0030] 8) optionally substituted heteroaralkyl;

[0031] Alternatively, R1 and R2 together with the nitrogen atom to which they are attached form an optionally substituted 3 to 8 membered heterocyclic ring;

[0032] R3 is hydroxy or fluorine;

[0033] R4 is selected from:

[0034] 1) Hydrogen;

[0035] 2) optionally substituted -C1-C6 alkyl;

[0036] 3) optionally substituted -C3-C8 cycloalkyl;

[0037] 4) an optionally substituted 3- to 8-membered heterocyclic ring;

[0038] 5) optionally substituted aryl;

[0039] 6) optionally substituted aralkyl;

[0040] 7) optionally substituted heteroaryl; and

[0041] 8) optionally substituted heteroaralkyl;

[0042] Each of the above preferred groups may be combined with one, any or all of the other preferred groups. DETAILED DESCRIPTION

[0043] One embodiment of the present invention is a compound of formula (I) as described above or a pharmaceutically acceptable salt thereof.

[0044] In certain embodiments of the compound of Formula (I), R1 is hydrogen.

[0045] In certain embodiments of the compound of formula (I), R1 is hydrogen and R2 is hydrogen.

[0046] In certain embodiments of the compound of Formula (I), R3 is -OH.

[0047] In certain embodiments of the compound of Formula (I), R4 is optionally substituted methyl or optionally substituted cyclopropyl.

[0048] In certain embodiments of the compound of formula (I), R4 is selected from one of the following:

[0049]

[0050] In certain embodiments of the compound of formula (I), R3 is OH and R4 is CF3, CD3 or cyclopropyl.

[0051] In certain embodiments of the compound of formula (I), R1 is hydrogen, R2 is hydrogen, R3 is OH, and R4 is CF3, CD3 or cyclopropyl.

[0052] In certain embodiments of the compound of formula (I), A is selected from one of the following with the removal of a hydrogen atom:

[0053]

[0054] wherein each of these groups is optionally substituted.

[0055] In certain embodiments of the compound of formula (I), A is selected from the following groups,

[0056]

[0057] In certain embodiments of the compound of formula (I), E is optionally substituted aryl, preferably optionally substituted phenyl.

[0058] In certain embodiments of the compound of formula (I), E is selected from the following groups,

[0059]

[0060] In certain embodiments of the compound of Formula (I), R4 is optionally substituted phenyl.

[0061] In certain embodiments of the compound of Formula (I), R4 is optionally substituted heteroaryl.

[0062] In certain embodiments of the compound of formula (I), R4 is selected from one of the following:

[0063]

[0064] Where R 31 Halogen, -CN, -OR 33 、-CO2R 33 、-SO2R 33 、-SO2NR 33 R 34 、-NR 33 R 34 , optionally substituted -C1-C6 alkyl, optionally substituted -C2-C6 alkenyl or optionally substituted -C3-C8 cycloalkyl; n is 0, 1 or 2; R33 and R34 are each independently selected from hydrogen, optionally substituted -C1-C6 alkyl, optionally substituted -C2-C6 alkenyl, optionally substituted -C3-C8 cycloalkyl, optionally substituted 3 to 8 membered heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl. Preferably, R 31 is selected from halogen, optionally substituted methyl and optionally substituted methoxy. More preferably, R 31 It is -F, -Cl, -CH3, -CHF2, -CF3 or -OCH3.

[0065] In certain embodiments of the compound of formula (I), R3 is OH and R4 is optionally substituted phenyl.

[0066] In certain embodiments of the compound of formula (I), A is selected from one of the following with the removal of a hydrogen atom: wherein each of these groups is optionally substituted.

[0067] In certain embodiments of the compounds of Formula (I), A is selected from

[0068]

[0069] In one embodiment of the present invention, the compound of formula (I) is represented by formula (Ia) or formula (Ib),

[0070]

[0071] wherein A, E, R1, R2, R3 and R4 are as described above. Preferably, the compound of formula (I) is represented by formula (Ia).

[0072] In one embodiment of the present invention, the compound of formula (I) is represented by formula (II),

[0073]

[0074] wherein A, E, R3 and R4 are as described above.

[0075] In one embodiment of the present invention, the compound of formula (I) is represented by formula (IIa),

[0076]

[0077] wherein A, E, R3 and R4 are as described above.

[0078] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (III-1) to (III-7),

[0079]

[0080] wherein A, E, R1 and R2 are as described above.

[0081] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (IV-1) to (IV-7),

[0082]

[0083] Wherein A and E are as described above.

[0084] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (IV-1) to (IV-7), and A is selected from the following groups,

[0085]

[0086] E is selected from the following groups,

[0087]

[0088] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (IV-1) to (IV-7), A is selected from one of the following except for the hydrogen atom, and each of these groups is optionally substituted:

[0089] E is selected from the following groups,

[0090]

[0091] In one embodiment of the present invention, the compound of formula (I) is represented by formula (V-1) or formula (V-2),

[0092]

[0093] wherein E and R4 are as described above, and

[0094] R 11 is selected from optionally substituted -C1-C6 alkyl, optionally substituted -C3-C8 cycloalkyl, optionally substituted 3 to 8 membered heterocycle, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl and optionally substituted heteroaralkyl. Preferably, R 11 is optionally substituted methyl or optionally substituted cyclopropyl. Preferably, R 11 is methyl or cyclopropyl.

[0095] R 12 is selected from hydrogen, optionally substituted -C1-C6 alkyl, optionally substituted -C3-C8 cycloalkyl, optionally substituted 3 to 8 membered heterocycle, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl and optionally substituted heteroaralkyl. Preferably, R 12 is optionally substituted methyl or optionally substituted cyclopropyl. More preferably, R 12 It is -CH3, -CHF2, -CD3, cyclopropyl,

[0096] R 13 and R 14 are each independently selected from hydrogen, halogen, -OR 11 , -NH2, optionally substituted -C1-C6 alkyl, optionally substituted -C3-C8 cycloalkyl, optionally substituted 3 to 8 membered heterocycle, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl and optionally substituted heteroaralkyl. Or, R 13 and R 14 Together with the carbon atoms to which they are attached, they form a 4 to 7 membered ring fused to the benzene ring. 13 and R 13 Each is independently selected from halogen, optionally substituted methyl and optionally substituted methoxy.

[0097] In one embodiment of the present invention, the compound of formula (I) is represented by formula (V-1) or formula (V-2), wherein E is a substituted phenyl group, and R4 is selected from one of the following:

[0098]

[0099] In one embodiment of the present invention, the compound of formula (I) is represented by formula (V-1) or formula (V-2), wherein E is selected from the following groups,

[0100]

[0101] In one embodiment of the present invention, the compound of formula (I) is represented by formula (V-1) or formula (V-2), wherein E is a substituted phenyl group; R 11 is optionally substituted methyl or optionally substituted cyclopropyl; R 12 is optionally substituted methyl or optionally substituted cyclopropyl; R 13 and R 14 Each is independently selected from halogen, optionally substituted methyl and optionally substituted methoxy; R4 is selected from one of the following:

[0102]

[0103] In one embodiment of the present invention, the compound of formula (I) is represented by formula (VI-1) or formula (VI-2),

[0104]

[0105] Among them, R4, R 11 , R 12 , R 13 and R 14 As mentioned above, and for each R 21 , R 22 , R 23 , R 24 and R 25 are independently selected from hydrogen, halogen, optionally substituted methyl and optionally substituted methoxy.

[0106] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (VII-1) to (VII-6),

[0107]

[0108] Where R 11 , R 12 , R 13 , R 14 , R 21 , R 22 , R 23 , R 24 and R 25 As mentioned before.

[0109] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (VIII-1) to (VIII-2),

[0110]

[0111] Among them, A, E, R1, R2, R 31 and n as described above.

[0112] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (VIII-1a) to (VIII-2a),

[0113]

[0114] Among them, A, E, R1, R2, R 31 and n as described above.

[0115] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (VIII-1a) to (VIII-2a), A is selected from one of the following except for the hydrogen atom, and each of these groups is optionally substituted:

[0116]

[0117] Wherein E is selected from the following groups,

[0118]

[0119] Preferably, R1 is hydrogen and R2 is hydrogen.

[0120] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (IX-1) to (IX-8),

[0121]

[0122] Among them, E and R 11 , R 12 , R 13 , R 14 , R 31 and n as described above.

[0123] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulae (IX-1a) to (IX-8a),

[0124]

[0125] Among them, E and R 11 , R 12 , R 13 , R 14 , R 31 and n as described above.

[0126] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (IX-1a) to (IX-8a), and E is selected from the following groups,

[0127]

[0128] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (X-1) to (X-12),

[0129]

[0130] Among them, E and R 31 and n as described above.

[0131] In one embodiment of the present invention, the compound of formula (I) is represented by one of formulas (X-1) to (X-12), and E is selected from the following groups,

[0132]

[0133] It should be understood that the description of the invention herein should be consistent with the laws and principles of chemical bonding. In some cases, it may be necessary to remove hydrogen atoms in order to accommodate a substituent at any given position.

[0134] It is intended that the definition of any substituent or variable (eg, R1, R2, etc.) at a particular location in a molecule be independent of its definitions elsewhere in the molecule.

[0135] It should be understood that compounds of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic, diastereoisomer and optically active forms. It should still be understood that some compounds of the present invention may exist in different tautomeric forms. All tautomers are considered to be within the scope of the present invention.

[0136] In certain embodiments, the present invention provides a method for preventing or treating RSV activity, and a method for treating RSV infection in a subject in need thereof. The method comprises administering to the subject an effective therapeutic amount of a compound of formula (I).

[0137] The present invention also provides use of the compound of formula (I) in preparing a drug for preventing or treating RSV.

[0138] Therefore, in one embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is combined with a steroidal anti-inflammatory compound, such as budesonide or fluticasone. In a preferred embodiment, the steroid is administered at a low dose to minimize immunosuppressive effects. In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is combined with a non-steroidal anti-inflammatory compound, such as a leukotriene antagonist, such as Singulair (Merck) or Astra Zeneca, a phosphodiesterase 4 inhibitor, such as roflumilast (Altana), a TNF-α inhibitor, such as Enbrel (Amgen), infliximab (Centocor), adalimumab (Abbott) or CDP870 (Celltech) or NSAIDS. In another embodiment, the compound of formula (I) is combined with an interleukin 8 or interleukin 9 inhibitor. Therefore, the present invention also relates to a product containing a compound of formula (I) or a pharmaceutically acceptable salt thereof and an anti-inflammatory compound for simultaneous, separate or sequential use in the treatment of RSV.

[0139] The present invention further relates to a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof and an anti-influenza compound, and the use of this combination in treating accompanying RSV and influenza infections. Therefore, the present invention further relates to a product containing a compound of formula (I) or a pharmaceutically acceptable salt thereof and an anti-influenza compound, for simultaneously, individually or sequentially treating accompanying RSV and influenza infections. The compounds of the present invention can be administered in a variety of dosage forms. Therefore, they can be administered orally, for example, as tablets, lozenges, lozenges, aqueous or oily suspensions, dispersible powders or granules. The compounds of the present invention can also be administered parenterally, whether subcutaneously, intravenously, intramuscularly, intrasternally, transdermally or by infusion techniques. These compounds can also be administered as suppositories.

[0140] In one embodiment, the compounds of the invention are administered intranasally or intrabronchially. The invention also provides an inhaler or nebulizer containing a medicament comprising (a) a derivative of formula (I) as described above or a pharmaceutically acceptable salt thereof, and (b) a pharmaceutically acceptable carrier or diluent.

[0141] The present invention also provides a pharmaceutical composition, which contains the benzodiazepine derivative or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

[0142] The compounds of the present invention are generally formulated for administration with a pharmaceutically acceptable carrier or diluent. For example, a solid oral form may contain an active compound, which is a diluent such as lactose, glucose, sucrose, cellulose, corn starch or potato starch; a lubricant such as silicon dioxide, talc, stearic acid, magnesium stearate or calcium stearate and / or polyethylene glycol; a binder such as starch, gum arabic, gelatin, methylcellulose, carboxymethylcellulose or polyvinylpyrrolidone; a disintegrant such as starch, alginic acid, alginate or sodium starch glycolate; an effervescent compound; a dye; a sweetener; a wetting agent such as lecithin, polysorbate, lauryl sulfate; and a non-toxic and pharmacologically inactive substance commonly used in pharmaceutical preparations. Such pharmaceutical preparations can be manufactured in a known manner, for example, by mixing, granulating, tableting, sugar coating or film coating processes.

[0143] Liquid dispersions for oral administration may be syrups, emulsions and suspensions. Syrups may contain carriers such as sucrose or sucrose with glycerol and / or mannitol and / or sorbitol.

[0144] Suspensions and emulsions may contain carriers such as natural gum, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose or polyvinyl alcohol. Suspensions or solutions for intramuscular injection may contain the active compound and a pharmaceutically acceptable carrier such as sterile water, olive oil, ethyl oleate, glycols such as propylene glycol, and, if desired, an appropriate amount of lidocaine hydrochloride.

[0145] Solutions for injection or infusion may contain a carrier, such as sterile water, or, preferably, they may be in the form of sterile, aqueous, isotonic saline solutions.

[0146] The present invention also relates to a method for treating a human or animal body using the novel compound or a pharmaceutically acceptable salt thereof as described above. The present invention also relates to a pharmaceutical composition comprising the novel compound as described above and a pharmaceutically acceptable diluent or carrier. Preferably, the pharmaceutical composition comprises a pharmaceutically acceptable salt of the novel compound as described above. Pharmaceutically acceptable salts are as defined above. The novel compound of the present invention is usually administered in the manner described above, and the compound is usually formulated for administration in the manner described above.

[0147] Preferably, the pharmaceutical composition comprises an optical isomer of the novel compound of the present invention. Thus, for example, preferred novel compounds of the present invention containing only one chiral center include substantially pure R enantiomers, substantially pure S enantiomers, and enantiomeric mixtures containing excess R enantiomers or excess S enantiomers. It is particularly preferred that the medicament contains substantially pure optical isomers of the compounds of the present invention. For the avoidance of doubt, the novel compounds of the present invention may be used in the form of solvates, if desired.

[0148] Another aspect of the invention is a method of preparing any of the compounds described herein using any of the synthetic means described herein.

[0149] Definition

[0150] Listed below are definitions of various terms used to describe the present invention. These definitions apply to the terms as used in this specification and claims, unless otherwise limited in specific instances, whether used individually or as part of a larger group.

[0151] The term "aryl" as used herein refers to a monocyclic, bicyclic or polycyclic carbocyclic ring system comprising at least one aromatic ring, including but not limited to phenyl, naphthyl, tetrahydronaphthyl, indanyl and indenyl. Polycyclic aryl is a polycyclic ring system comprising at least one aromatic ring. Polycyclic aryl can include fused rings, covalently linked rings or a combination thereof.

[0152] The term "heteroaryl" as used herein refers to a mono-, bi- or polycyclic aromatic group having one or more ring atoms selected from S, O and N; and the remaining ring atoms are carbon, wherein any N or S contained in the ring may be optionally oxidized. Heteroaryl includes, but is not limited to, pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, thienyl, furanyl, quinolyl, isoquinolyl, benzimidazolyl, benzoxazolyl, quinoxalinyl. Polycyclic heteroaryl may include fused rings, covalently linked rings or combinations thereof.

[0153] According to the invention, the aromatic groups may be substituted or unsubstituted.

[0154] The term "bicyclic aryl" or "bicyclic heteroaryl" refers to a ring system consisting of two rings, at least one of which is aromatic; and the two rings may be fused or covalently linked.

[0155] The term "alkyl" as used herein refers to a saturated straight or branched hydrocarbon group. "C1-C3 alkyl", "C1-C6 alkyl", "C1-C 10 "C-C alkyl", "C2-C4 alkyl" or "C3-C6 alkyl" refers to an alkyl group containing 1 to 3, 1 to 6, 1 to 10 carbon atoms, 2 to 4 and 3 to 6 carbon atoms, respectively. Examples of C1-C8 alkyl include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl and octyl.

[0156] As used herein, the term "alkenyl" refers to a straight or branched chain hydrocarbon group having at least one carbon-carbon double bond by removing a single hydrogen atom. 10"C2-C8 alkenyl", "C2-C4 alkenyl" or "C3-C6 alkenyl" refers to an alkenyl group containing 2 to 10, 2 to 8, 2 to 4 or 3 to 6 carbon atoms, respectively. Alkenyl includes, but is not limited to, for example, vinyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, heptenyl, octenyl, etc.

[0157] As used herein, the term "alkynyl" refers to a straight or branched chain hydrocarbon group having at least one carbon-carbon triple bond by removing a single hydrogen atom. 10 "C2-C8 alkynyl", "C2-C4 alkynyl" or "C3-C6 alkynyl" refers to an alkynyl group containing 2 to 10, 2 to 8, 2 to 4 or 3 to 6 carbon atoms, respectively. Representative alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl, and the like.

[0158] The term "cycloalkyl" as used herein refers to a monocyclic or polycyclic saturated carbocyclic or bicyclic or tricyclic group of a fused, bridged or spirocyclic system, the carbon atoms of which may be optionally substituted by oxo or optionally substituted by an exocyclic olefin, imine or oxime double bond. Preferred cycloalkyl groups include C3-C 12 Cycloalkyl, C3-C6 cycloalkyl, C3-C8 cycloalkyl and C4-C7 cycloalkyl. 12 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl, cyclooctyl, 4-methylenecyclohexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.0]hexyl, spiro[2.5]octyl, 3-methylenebicyclo[3.2.1]octyl, spiro[4.4]nonyl, and the like.

[0159] As used herein, the term "cycloalkenyl" refers to a monocyclic or polycyclic carbocyclic or bicyclic or tricyclic group of a fused, bridged or spiro ring system having at least one carbon-carbon double bond, and the carbon atoms may be optionally substituted by oxo or optionally substituted by an exocyclic olefin, imine or oxime double bond. Preferred cycloalkenyl groups include C3-C 12 Cycloalkenyl, C3-C8 cycloalkenyl or C5-C7 cycloalkenyl. 12 Examples of cycloalkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cyclooctenyl, bicyclo[2.2.1]hept-2-enyl, bicyclo[3.1.0]hex-2-enyl, spiro[2.5]oct-4-enyl, spiro[4.4]non-1-enyl, bicyclo[4.2.1]non-3-en-9-yl, and the like.

[0160] As used herein, the term "aralkyl" refers to a functional group in which an alkylene chain is attached to an aryl group, for example -CH2CH2-phenyl. The term "substituted aralkyl" refers to an arylalkyl functional group in which an aryl group is substituted. Similarly, the term "heteroaralkyl" refers to a functional group in which an alkylene chain is attached to a heteroaryl group. The term "substituted heteroarylalkyl" refers to a heteroarylalkyl functional group in which a heteroaryl group is substituted.

[0161] As used herein, unless otherwise indicated, the term "alkoxy", used alone or in combination with other terms, refers to an alkyl group having the specified number of carbon atoms, which is attached to the remainder of the molecule via an oxygen atom, such as methoxy, ethoxy, 1-propoxy, 2-propoxy (isopropoxy) and higher homologs and isomers. A preferred alkoxy group is (C1-C3)alkoxy.

[0162] It is to be understood that any alkyl, alkenyl, alkynyl, cycloalkyl, heterocycle and cycloalkenyl moieties described herein may also be an aliphatic or alicyclic group.

[0163] An "aliphatic" group is a non-aromatic moiety composed of any combination of carbon atoms, hydrogen atoms, halogen atoms, oxygen, nitrogen or other atoms, and optionally contains one or more unsaturated units, such as double and / or triple bonds. Examples of aliphatic groups are functional groups such as alkyl, alkenyl, alkynyl, O, OH, NH, NH2, C(O), S(O)2, C(O)O, C(O)NH, OC(O)O, OC(O)NH, OC(O)NH2, S(O)2NH, S(O)2NH2, NHC(O)NH2, NHC(O)C(O)NH, NHS(O)2NH, NHS(O)2NH2, C(O)NHS(O)2, C(O)NHS(O)2NH or C(O)NHS(O)2NH2, etc., groups containing one or more functional groups, non-aromatic hydrocarbons (optionally substituted) and groups in which one or more carbons of the non-aromatic hydrocarbons (optionally substituted) are substituted with functional groups. The carbon atoms of the aliphatic group may optionally be substituted with oxo. The aliphatic group may be straight chain, branched, cyclic or a combination thereof, preferably containing from about 1 to about 24 carbon atoms, more typically containing from about 1 to about 12 carbon atoms. In addition to the aliphatic hydrocarbon groups used herein, aliphatic groups explicitly include, for example, alkoxyalkyl, polyalkoxyalkyl, such as polyalkylene glycols, polyamines and polyimines. The aliphatic group may optionally be substituted.

[0164] The term "carbocycle" or "carbocyclic" refers to a saturated, partially unsaturated or aromatic cyclic group in which every atom in the ring is carbon. Examples of heterocyclic compounds include cycloalkyl, cycloalkenyl and aryl groups.

[0165] The terms "heterocycle" or "heterocycloalkyl" are used interchangeably and refer to a non-aromatic ring or a bicyclic or tricyclic radical of a fused, bridged or spiro ring system wherein (i) each ring system contains at least one heteroatom independently selected from oxygen, sulfur and nitrogen, (ii) each ring system may be saturated or unsaturated, (iii) the nitrogen and sulfur heteroatoms may optionally be oxidized, (iv) the nitrogen heteroatom may optionally be quaternized, (v) any of the above rings may be fused to an aromatic ring, and (vi) the remaining ring atoms are carbon atoms which may be optionally oxo-substituted or optionally substituted with an exocyclic olefin, imine or oxime double bond. Representative heterocycloalkyl includes but is not limited to 1,3-dioxolane, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, 2-azabicyclo [2.2.1] -heptyl, 8-azabicyclo [3.2.1] octyl, 5-azaspiro [2.5] octyl, 1-oxa -7-azaspiro [4.4] nonyl, 7-oxooxacycloheptan-4-yl and tetrahydrofuranyl. These heterocyclic groups can be further substituted. Heteroaryl or heterocyclic groups can be C- connected or N- connected (where possible).

[0166] It should be understood that any alkyl, alkenyl, alkynyl, alicyclic, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycle, aliphatic moiety, etc. described herein can also be a divalent or multivalent group when used as a bond connecting two or more groups or substituents, which can be on the same or different atoms. Those skilled in the art can easily determine the valence of any such group based on the context in which it appears.

[0167] The term "substituted" refers to substitution by independently replacing one, two, three or more hydrogen atoms with substituents, including but not limited to -F, -Cl, -Br, -I, -OH, -C1-C 12 Alkyl, -C2-C 12 Alkenyl, -C2-C 12 Alkynyl, -C3-C 12 Cycloalkyl, protected hydroxy, -NO2, -N3, -CN, -NH2, protected amino, oxo, thio, -NH-C1-C 12 Alkyl, -NH-C2-C8 alkenyl, -NH-C2-C8 alkynyl, -NH-C3-C 12 Cycloalkyl, -NHaryl, -NHheteroaryl, -N-heterocycloalkyl, -dialkylamino, -diarylamino, -bisheteroarylamino, -O-C1-C 12 Alkyl, -O-C2-C8 alkenyl, -O-C2-C8 alkynyl, -O-C3-C 12Cycloalkyl, -O-aryl, -O-heteroaryl, -O-heterocycloalkyl, -C(O)-C1-C 12 alkyl, -C(O)-C2-C8 alkenyl, -C(O)C2-C8 alkynyl, -C(O)C3-C8 cycloalkyl, -C-(O)-aryl, -C(O)-heteroaryl, -C(O)-heterocycloalkyl, -CONH2, -CONH-C1-C8 12 -alkyl, -CONH-C2-C8-alkenyl, -CONH-C2-C8-alkynyl, -CONH-C3-C 12 -cycloalkyl, -CONH-aryl, -CONH-heteroaryl, -CONH-heterocycloalkyl, -OCO2-C1-C 12 -alkyl, -OCO2-C2-C8-alkenyl, -OCO2-C2-C8-alkynyl, -OCO2-C3-C 12 -cycloalkyl, -OCO2-aryl, -OCO2-heteroaryl, -OCO2-heterocycloalkyl, -CO2-C1-C 12 Alkyl, -CO2-C2-C8 alkenyl, -CO2-C2-C8 alkynyl, CO2-C3-C 12 -cycloalkyl, -CO2-aryl, CO2-heteroaryl, CO2-heterocycloalkyl, -OCONH2, -OCONH-C1-C 12 -alkyl, -OCONH-C2-C8-alkenyl, -OCONH-C2-C8-alkynyl, -OCONH-C3-C 12 -cycloalkyl, -OCONH-aryl, -OCONH-heteroaryl, -OCONH-heterocyclo-alkyl, -NHC(O)H, -NHC(O)-C1-C 12 -alkyl, -NHC(O)-C2-C8-alkenyl, -NHC(O)-C2-C8-alkynyl, -NHC(O)-C3-C 12 -cycloalkyl, -NHC(O)-aryl, -NHC(O)-heteroaryl, -NHC(O)-heterocyclo-alkyl, -NHCO2-C1-C 12 -alkyl, -NHCO2-C2-C8-alkenyl, -NHCO2-C2-C8-alkynyl, -NHCO2-C3-C 12 -cycloalkyl, -NHCO2-aryl, -NHCO2-heteroaryl, -NHCO2-heterocycloalkyl, -NHC(O)NH2, -NHC(O)NH-C1-C 12 -alkyl, -NHC(O)NH-C2-C8-alkenyl, -NHC(O)NH-C2-C8-alkynyl, -NHC(O)NH-C3-C 12-cycloalkyl, -NHC(O)NH-aryl, -NHC(O)NH-heteroaryl, -NHC(O)NH-heterocycloalkyl, NHC(S)NH2, -NHC(S)NH-C1-C 12 -alkyl, -NHC(S)NH-C2-C8-alkenyl, -NHC(S)NH-C2-C8-alkynyl, -NHC(S)NH-C3-C 12 -cycloalkyl, -NHC(S)NH-aryl, -NHC(S)NH-heteroaryl, -NHC(S)NH-heterocycloalkyl, -NHC(NH)NH2, -NHC(NH)NH-C1-C 12 -alkyl, -NHC(NH)NH-C2-C8-alkenyl, -NHC(NH)NH-C2-C8-alkynyl, -NHC(NH)NH-C3-C 12 -cycloalkyl, -NHC(NH)NH-aryl, -NHC(NH)NH-heteroaryl, -NHC(NH)NH-heterocycloalkyl, -NHC(NH)-C1-C 12 -alkyl, -NHC(NH)-C2-C8-alkenyl, -NHC(NH)-C2-C8-alkynyl, -NHC(NH)-C3-C 12 -cycloalkyl, -NHC(NH)-aryl, -NHC(NH)-heteroaryl, -NHC(NH)-heterocycloalkyl, -C(NH)NH-C1-C 12 -alkyl, -C(NH)NH-C2-C8-alkenyl, -C(NH)NH-C2-C8-alkynyl, -C(NH)NH-C3-C 12 -cycloalkyl, -C(NH)NH-aryl, -C(NH)NH-heteroaryl, -C(NH)NH-heterocycloalkyl, -S(O)-C1-C 12 -alkyl, -S(O)-C2-C8-alkenyl, -S(O)-C2-C8-alkynyl, -S(O)-C3-C 12 -cycloalkyl, -S(O)-aryl, -S(O)-heteroaryl, -S(O)-heterocycloalkyl, -SO2NH2, -SO2NH-C1-C 12 -alkyl, -SO2NH-C2-C8-alkenyl, -SO2NH-C2-C8-alkynyl, -SO2NH-C3-C 12 -cycloalkyl, -SO2NH-aryl, -SO2NH-heteroaryl, -SO2NH-heterocycloalkyl, -NHSO2-C1-C 12 -alkyl, -NHSO2-C2-C8-alkenyl, -NHSO2-C2-C8-alkynyl, -NHSO2-C3-C 12-cycloalkyl, -NHSO2-aryl, -NHSO2-heteroaryl, -NHSO2-heterocycloalkyl, -CH2NH2, -CH2SO2CH3, -aryl, -arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl, -C3-C 12 -cycloalkyl, polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, -SH, -S-C1-C 12 -alkyl, -S-C2-C8-alkenyl, -S-C2-C8-alkynyl, -S-C3-C 12 -cycloalkyl, -S-aryl, -S-heteroaryl, -S-heterocycloalkyl or methylthiomethyl. In certain embodiments, the substituent is independently selected from halogen, preferably Cl and F; C1-C4-alkyl, preferably methyl and ethyl; halogenated C1-C4 alkyl, such as fluoromethyl, difluoromethyl and trifluoromethyl; C2-C4-alkenyl; halogenated-C2-C4-alkenyl; C3-C6-cycloalkyl, such as cyclopropyl; C1-C4-alkoxy, such as methoxy and ethoxy; halogenated-C1-C4-alkoxy, such as fluoromethoxy, difluoromethoxy and trifluoromethoxy, -CN; -OH; NH2; C1-C4-alkylamino; di(C1-C4-alkyl)amino; and NO2. It should be understood that aryl, heteroaryl, alkyl, etc. can be further substituted. In some cases, when possible, each substituent in a substituted moiety is further optionally substituted with one or more groups, each group independently selected from C1-C4-alkyl, -CF3, -OCH3, -OCF3, -F, -Cl, -Br, -I, -OH, -NO2, -CN and -NH2.

[0168] In certain embodiments, the substituted alkyl, alkenyl or alkoxyl groups are substituted with one or more halogen atoms, preferably fluorine or chlorine atoms. Such substituted alkyl groups include fluoromethyl, difluoromethyl and trifluoromethyl. Such substituted alkoxyl groups include fluoromethoxy, difluoromethoxy and trifluoromethoxy.

[0169]

[0046] The terms "halo" or "halogen," as used herein, alone or as part of another substituent, refer to a fluorine, chlorine, bromine, or iodine atom.

[0170] As used herein, the term "optionally substituted" means that the reference group can be substituted or unsubstituted. In one embodiment, the reference group is optionally substituted with zero substituents, i.e., the reference group is unsubstituted. In another embodiment, the reference group is optionally substituted with one or more additional groups individually and independently selected from the groups described herein.

[0171] The term "hydrogen" includes hydrogen and deuterium. In addition, recitations of atoms include other isotopes of that atom as long as the resulting compound is pharmaceutically acceptable.

[0172] In certain embodiments, the compound of each formula herein is defined as including isotope-labeled compounds. "Isotope-labeled compound" is a compound in which at least one atomic position is enriched in a specific isotope of a specified element, and its enrichment level is significantly higher than the natural abundance of the isotope. For example, one or more hydrogen atom positions in the compound can be enriched with deuterium to a level significantly greater than the natural abundance of deuterium, for example, enriched to at least 1%, preferably at least 20% or at least 50%. For example, this deuterated compound may be metabolized more slowly than its non-deuterated analog, and therefore shows a longer half-life when administered to a subject. This compound can be synthesized using methods known in the art, for example, by using deuterated raw materials. Unless otherwise indicated, isotope-labeled compounds are pharmaceutically acceptable.

[0173] The term "hydroxyl activating group" as used herein refers to an unstable chemical moiety known in the art that activates a hydroxyl group, allowing it to be separated during a synthetic process such as a substitution or elimination reaction. Examples of hydroxyl activating groups include, but are not limited to, methanesulfonates, toluenesulfonates, trifluoromethanesulfonates, p-nitrobenzoates, phosphonates, and the like.

[0174] As used herein, the term "activated hydroxyl" refers to a hydroxyl group activated with a hydroxyl activating group, as described above, including mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups.

[0175] The term "hydroxy protecting group" as used herein refers to a labile chemical moiety known in the art that is used to protect a hydroxyl group from undesirable reactions during synthesis. After the synthesis steps described, the hydroxy protecting groups described herein can be selectively removed. Hydroxyl protecting groups known in the art are generally described in TH Greene and PGM Wuts, Protective Groups in Organic Synthesis , 3rd edition, John Wiley & Sons, New York (1999). Examples of hydroxyl protecting groups include benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, tert-butyloxycarbonyl, isopropoxycarbonyl, diphenylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, tert-butyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, allyl, benzyl, triphenylmethyl (trityl), methoxymethyl, methylthiomethyl, benzyloxymethyl, 2-(trimethylsilyl)-ethoxymethyl, methylsulfonyl, trimethylsilyl, triisopropylsilyl, and the like.

[0176] The term "protected hydroxy" as used herein refers to a hydroxy group protected with a hydroxy protecting group, as described above, including benzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl.

[0177] The term "hydroxy prodrug group" as used herein refers to a primary group known in the art that changes the physicochemical properties of the parent drug in a transient manner by covering or masking the hydroxyl group, thereby changing its biological properties. After the synthetic procedure, the hydroxy prodrug group described herein must be able to revert to the hydroxyl group in vivo. Hydroxyl prodrug groups known in the art are generally described in Kenneth B. Sloan, Prodrugs, Topical and Ocular Drug Delivery , (Drugs and the Pharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York (1992) and "Prodrugs of Alcohols and Phenols" by SSDhareshwar and VJStella, Prodrugs Challenges and Rewards Part - 2 (Biotechnology: Pharmaceutical Aspects), edited by V. J. Stella et al., Springer and AAPS Press, 2007, pp 31-99.

[0178] As used herein, the term "amino protecting group" refers to a labile chemical moiety known in the art that is used to protect an amino group from undesirable reactions during a synthetic process. After the synthetic process, the amino protecting group described herein can be selectively removed. Amino protecting groups known in the art are generally described in TH Greene and PGM Wuts, Protective Groups in Organic Synthesis , 3rd edition, John Wiley & Sons, New York (1999). Examples of amino protecting groups include, but are not limited to, methoxycarbonyl, tert-butyloxycarbonyl, 9-fluorenyl-methoxycarbonyl, benzyloxycarbonyl, and the like.

[0179] The term "protected amino group" as used herein refers to an amino group protected with an amino protecting group as described above.

[0180] The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom in a substitution reaction (such as a nucleophilic substitution reaction). For example, representative leaving groups include chloro, bromo and iodo groups; sulfonate groups, such as mesylate, tosylate, p-brosylate, nitrobenzenesulfonate, etc.; and acyloxy groups, such as acetoxy, trifluoroacetoxy, etc.

[0181] As used herein, the term "aprotic solvent" refers to a solvent that is relatively inert to proton activity, i.e., does not act as a proton donor. Examples include, but are not limited to, hydrocarbons such as hexane and toluene, halogenated hydrocarbons such as dichloromethane, vinyl chloride, chloroform, etc., heterocyclic compounds such as tetrahydrofuran and N-methylpyrrolidone, and ethers such as diethyl ether and bismethoxymethyl ether. These compounds are well known to those skilled in the art, and it is obvious to those skilled in the art that for specific compounds and reaction conditions, a single solvent or a mixture thereof may be preferred, depending on factors such as the solubility of the reagents, the reactivity of the reagents, and the preferred temperature range. Further discussion of aprotic solvents can be found in organic chemistry textbooks or professional monographs, for example: Organic Solvents Physical Properties and Methods of Purification ,4th ed.,edited byJohn A. Riddick et al., Vol.II,in the Techniques of Chemistry Series ,John Wiley&Sons,NY,1986.

[0182] As used herein, the term "protic solvent" refers to a solvent that tends to donate protons, such as alcohols, for example, methanol, ethanol, propanol, isopropanol, butanol, tert-butanol, etc. These solvents are well known to those skilled in the art, and it is obvious to those skilled in the art that for specific compounds and reaction conditions, a single solvent or a mixture thereof may be preferred, depending on factors such as the solubility of the reagents, the reactivity of the reagents, and the preferred temperature range. Further discussion of protic solvents can be found in organic chemistry textbooks or professional monographs, for example: Organic Solvents Physical Pro perties and Methods of Purification ,4th ed.,edited by John A. Riddick et al.,Vol.II,in the Techniques of Chemistry Series ,John Wiley & Sons,NY,1986.

[0183] Combinations of substituents and variables contemplated by this invention are only those that result in the formation of stable compounds. As used herein, the term "stable" refers to compounds that are sufficiently stable to be manufactured and that maintain the integrity of the compound for a sufficient period of time to be useful for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

[0184] The synthesized compound can be isolated from the reaction mixture and further purified by column chromatography, high pressure liquid chromatography or recrystallization. It will be appreciated by those skilled in the art that further methods for synthesizing the compounds of the formula herein will be apparent to those of ordinary skill in the art. In addition, the various synthetic steps can be performed in an alternating sequence or order to obtain the desired compounds. Synthetic chemical transformations and protecting group methods (protection and deprotection) that can be used to synthesize the compounds described herein are known in the art, for example, as described in R. Larock, Comprehensive Organic Transf ormations ,2 nd Ed. Wiley-VCH (1999); TW Greene and PGMWuts, Protective Groups in Organic Synthesis ,3rdEd.,JohnWiley and Sons(1999);L.Fieser and M.Fieser, Fieser and Fieser's Reagents for Organic Synthesis , John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis , John Wiley and Sons (1995) and subsequent editions.

[0185] The term "subject" as used herein refers to an animal. Preferably, the animal is a mammal. More preferably, the mammal is a human. For example, a subject also refers to a dog, a cat, a horse, a cow, a pig, a guinea pig, a fish, a bird, etc.

[0186] The compounds of the present invention can be modified by adding appropriate functional groups to enhance selective biological properties. Such modifications are known in the art and may include those that increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow injection administration, alter metabolism, and alter excretion rate.

[0187] The compounds described herein contain one or more asymmetric centers and thus give rise to enantiomers, diastereomers, and other stereoisomeric forms. Based on absolute stereochemistry, they can be defined as the (R)- or (S)-, or (D)- / or (L)-forms of the amino acids. The present invention is intended to include all such possible isomers, as well as their racemic and optically pure forms. Optical isomers can be prepared by the above procedures or by resolving the racemic mixtures from their respective optically active precursors. The resolution can be carried out in the presence of a resolving agent, by chromatography or repeated crystallization, or by some combination of these techniques known to those skilled in the art. More details on the resolution can be found in Jacques et al., Enantiomers, Racemates, and Resolutions(John Wiley & Sons, 1981). When the compounds described herein contain olefinic double bonds, other unsaturation, or other centers of geometric asymmetry, unless otherwise specified, the compounds shall include E and Z geometric isomers or cis and trans isomers. Likewise, all tautomeric forms shall be included. Tautomers may be cyclic or acyclic. The configuration of any carbon-carbon double bond appearing herein is selected for convenience only and is not intended to specify a particular configuration unless otherwise specified in the text; therefore, any carbon-carbon double bond or carbon heteroatom double bond described herein as trans may be cis, trans, or a mixture of the two in any proportion.

[0188] Certain compounds of the present invention may also exist in different stable conformational forms that can be separated. Torsional asymmetry due to restricted rotation around an asymmetric single bond (e.g., due to steric hindrance or ring strain) may allow separation of different conformations. The present invention includes each conformational isomer of these compounds and mixtures thereof.

[0189] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that is suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., and has a reasonable benefit / risk ratio within the scope of reasonable medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). Salts can be prepared in situ during the final separation and purification of the compounds of the present invention, or they can be prepared separately by reacting the free base functional group with a suitable organic acid. Examples of pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts, which are salts formed by amino groups with inorganic or organic acids, or salts formed by using other methods used in the art, such as ion exchange, inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, and organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid. Other pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyls having 1 to 6 carbon atoms, sulfonates and aryl sulfonates, where appropriate.

[0190] Pharmaceutically acceptable salts can also be prepared by deprotonating the parent compound with a suitable base to form an anionic conjugate base of the parent compound. In such salts, the counterion is a cation. Suitable cations include ammonium and metal cations, such as alkali metal cations, including Li + 、Na + , K + and Cs + , and alkaline earth metal cations such as Mg 2+ and Ca2+ .

[0191] As used herein, the term "pharmaceutically acceptable ester" refers to esters that hydrolyze in vivo, including those that readily decompose in the human body to leave the parent compound or a salt thereof. Suitable ester groups include, for example, esters derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanoic acids, alkenoic acids, cycloalkanoic acids, and alkanedioic acids, wherein each alkyl or alkenyl moiety advantageously has no more than 6 carbon atoms. Examples of specific esters include, but are not limited to, C1-C6 alkanoic acid esters, such as acetate, propionate, butyrate, and pivalate.

[0192] In certain embodiments, the present invention provides pharmaceutically acceptable prodrugs of the compounds disclosed herein. The term "pharmaceutically acceptable prodrug" as used herein refers to a prodrug of a compound formed by the method of the present invention, which is suitable for contact with the tissues of humans and lower animals within the scope of reasonable medical judgment, has excessive toxicity, irritation, allergic reactions, etc., has a reasonable benefit / risk ratio, is effective for its intended use, and a zwitterionic form of the compound of the present invention (if possible). "Prodrug" as used herein refers to a compound that can be converted into any compound shown in the formula of the present invention in vivo by metabolic means (e.g., by hydrolysis). Various forms of prodrugs are known in the art and are described in, for example, Bundgaard, (ed.), Design of Prodrugs, Elsevier (1985); Widder et al. (ed.), Methods in Enzymology, Vol. 4, Academic Press (1985); Krogsgaard-Larsen et al. (ed.). "Design and Application of Prodrugs, Textbook of Drug Design and Development, Chapter 5, 113-191 (1991); Bundgaard et al., Journal of Drug Deliver Reviews, 8: 1-38 (1992); Bundgaard, J. of Pharmaceutical Sciences, 77: 285 et seq. (1988); Higuchi and Stella (eds.) Prodrugs as Novel Drug Delivery Systems, American Chemical Society (1975); and Bernard Testa & Joachim Mayer, "Hydrolysis In Drug And Prodrug Metabolism: Chemistry, Biochemistry and Enzymology,” John Wiley and Sons, Ltd. (2002).

[0193] Other types of prodrugs are also included. For example, free carboxyl groups can be derived into amides or alkyl esters. Free hydroxyl groups can be derivatized using groups including but not limited to hemisuccinate, ethyl succinate, phosphate, dimethylaminoacetate and phosphoryloxymethoxycarbonyl, as described in Advanced Drug Delivery Reviews, 1996, 19, 115. Also included are carbamate prodrugs of hydroxyl and amino groups, and carbonate prodrugs, sulfonates and sulfates of hydroxyl groups. Also included are hydroxyl groups derived into (acyloxy) methyl and (acyloxy) ethyl ethers, wherein the acyl group can be an alkyl ester, optionally substituted by groups including but not limited to ether, amine and carboxylic acid functional groups, or wherein the acyl group is an amino acid ester as described above. Such prodrugs are described in J.Med.Chem.1996, 39, 10. Free amines can also be derived into amides, sulfonamides or phosphonamides. All of these prodrug moieties can be combined with groups including but not limited to ether, amine and carboxylic acid functional groups. In certain embodiments, the compounds of the present invention can incorporate two or more groups that are metabolically removed in vivo to produce active parent compounds.

[0194] As used herein, the term "treat" means to alleviate, lessen, reduce, eliminate, modulate, or improve, i.e., to cause regression of a disease state or condition. Treatment may also include inhibition, i.e., arresting the development of an existing disease state or condition, as well as alleviation or improvement, i.e., causing regression of existing disease states and conditions, such as when a disease state or condition may already exist.

[0195] As used herein, the term "preventing" refers to completely or nearly completely preventing a disease state or condition from occurring in a patient or subject, especially when the patient or subject is susceptible to or at risk of contracting such disease state or condition.

[0196] In addition, the compounds of the present invention, such as salts of the compounds, may exist in hydrated or unhydrated (anhydrous) forms, and may also exist as solvates with other solvent molecules. Non-limiting examples of hydrates include monohydrates, dihydrates, etc. Non-limiting examples of solvates include ethanol solvates, acetone solvates, etc.

[0197] "Solvate" refers to a solvent addition form containing either stoichiometric or non-stoichiometric amounts of solvent. Some compounds tend to trap fixed molar ratios of solvent molecules in the crystalline solid state, thereby forming solvates. If the solvent is water, the solvate formed is a hydrate, and when the solvent is an alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more water molecules with a substance in which the water remains in its molecular state as H2O, and this combination is able to form one or more hydrates.

[0198] As used herein, the term "analog" refers to a compound that is structurally similar to another but has a slightly different composition (such as one atom replaced by an atom of a different element, or in the presence of a particular functional group, or one functional group replaced by another functional group). Thus, an analog is a compound that is similar or comparable in function and appearance to a reference compound.

[0199] Combinations of substituents and variables contemplated by this invention are only those that result in the formation of stable compounds. As used herein, the term "stable" refers to compounds that have sufficient stability to allow manufacture and maintain the integrity of the compound for a sufficient period of time to be useful for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

[0200] The synthesized compound can be isolated from the reaction mixture and further purified by methods such as column chromatography, high pressure liquid chromatography or recrystallization. In addition, the various synthetic steps can be performed in an alternating order or sequence to obtain the desired compound. In addition, the solvents, temperatures, reaction durations, etc. described herein are for illustrative purposes only, and variations in reaction conditions can produce the desired bridged macrocyclic products of the present invention. Synthetic chemical transformations and protecting group methods (protection and deprotection) that can be used to synthesize the compounds described herein include, for example, those described in R. Larock, Comprehensive Organic Transformat ions ,VCH Publishers(1989);TWGreene and PGMWuts, Protective Groups in Organic Synthesis ,2d.Ed.,JohnWiley and Sons(1991);L.Fieser and M.Fieser, Fieser and Fieser's Reagents for Organic Synthesis , John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis ,John Wiley and Sons (1995).

[0201] The compounds of the present invention can be modified by adding various functional groups through the synthetic means described herein to enhance selective biological properties. These modifications include increasing biological penetration to a given biological system (such as blood, lymphatic system, central nervous system), increasing oral availability, increasing solubility to allow injection administration, changing metabolism and changing excretion rate.

[0202] Drug Composition

[0203] The pharmaceutical composition of the present invention comprises an effective therapeutic amount of the compound of the present invention, which is formulated together with one or more pharmaceutically acceptable carriers. As used herein, the term "pharmaceutically acceptable carrier" refers to any type of non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary. Some examples of materials that can be used as pharmaceutically acceptable carriers are sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol and phosphate buffered solutions; and the composition may also contain other nontoxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants, according to the judgment of the formulator. The pharmaceutical compositions of this invention may be administered to humans and other animals orally, rectally, parenterally, intracisternal, intravaginal, intraperitoneal, topically (as powders, ointments or drops), buccally or as an oral or nasal spray.

[0204] The pharmaceutical composition of the present invention can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or by an implanted reservoir, preferably orally or by injection. The pharmaceutical composition of the present invention can contain any conventional non-toxic pharmaceutically acceptable carrier, adjuvant or solvent. In some cases, the pH value of the preparation can be adjusted with pharmaceutically acceptable acid, alkali or buffer to improve the stability of the preparation compound or its delivery form. The term parenteral injection used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.

[0205] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compound, the liquid dosage form may also contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oil (particularly cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and sorbitan fatty acid esters, and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and aromatics.

[0206] Injectable preparations, such as sterile injectable aqueous or oily suspensions, can be prepared using suitable dispersants or wetting agents and suspending agents according to known techniques. Sterile injectable preparations can also be sterile injection solutions, suspensions or emulsions, dissolved in nontoxic parenteral acceptable diluents or solvents, for example, dissolved in 1,3-butanediol. Available acceptable vehicles and solvents include water, Ringer's solution, USP and isotonic sodium chloride solution. In addition, sterile fixed oils are generally used as solvents or suspension media. For this reason, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid are used to prepare injections.

[0207] The injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0208] In order to prolong the effect of the drug, it is usually necessary to slow down the absorption of subcutaneous or intramuscular injection of the drug. This can be achieved by using a liquid suspension of a crystalline or amorphous material with poor water solubility. Then, the absorption rate of the drug depends on its dissolution rate, which in turn may depend on the crystal size and crystal form. Alternatively, delayed absorption of parenteral administration of the drug form is achieved by dissolving or suspending the drug in an oily solvent. The injectable reservoir form is made by forming a microcapsule matrix of the drug in a biodegradable polymer (such as polylactic acid-polyglycolide). Depending on the ratio of the drug to the polymer and the properties of the specific polymer used, the drug release rate can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injection preparations can also be prepared by embedding the drug in liposomes or microemulsions compatible with body tissues.

[0209] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

[0210] Orally administrable solid dosage forms include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate and / or: a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and silicic acid, b) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia, c) humectants, such as glycerol, d) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate, e) solution retarders, such as paraffin, f) absorption promoters, such as quaternary ammonium compounds, g) wetting agents, for example, cetyl alcohol and glyceryl monostearate, h) absorbents, such as kaolin and bentonite, i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

[0211] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols.

[0212] The active compound may also be in microcapsule form, as described above, containing one or more excipients. Solid dosage forms of tablets, dragees, capsules, pills and granules may be prepared with coatings and shells, such as enteric coatings, controlled release coatings and other coatings known in the field of pharmaceutical preparations. In such solid dosage forms, the active compound may be mixed with at least one inert diluent, such as sucrose, lactose or starch. Conventionally, such dosage forms may also include other substances other than inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage form may also include a buffer. They may optionally contain an opacifier, and may also have a composition that releases the active ingredient only or preferably in a delayed manner at a specific site in the intestinal tract. Examples of usable embedding compositions include polymers and waxes.

[0213] Topical or transdermal dosage forms of the compounds of the invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active ingredient is mixed under aseptic conditions with a pharmaceutically acceptable carrier and any desired preservatives or buffers that may be needed. Ophthalmic preparations, ear drops, eye ointments, powders and solutions are also considered within the scope of the present invention.

[0214] Ointments, pastes, creams and gels may contain, in addition to the active compounds of this invention, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0215] In addition to the compounds of this invention, powders and sprays may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate and polyamide powder, or mixtures of these substances. Sprays may also contain common propellants such as chlorofluorocarbons.

[0216] Transdermal patches have the additional advantage of allowing controlled delivery of compounds into the body. This dosage form can be prepared by dissolving or dispensing the compound into an appropriate medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0217] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by one of ordinary skill in the art. All publications, patents, published patent applications, and other references mentioned herein are incorporated herein by reference in their entirety.

[0218] Abbreviation

[0219] The abbreviations used in the protocol description and in the following examples are:

[0220] ACN stands for acetonitrile;

[0221] AD-mix- stands for (9S)-(9”S)-9,9”-[1,4-Phthalazinediylbis(oxy)]bis[10,11-dihydro-6'-methoxycinchonan];

[0222] Bn represents benzyl;

[0223] BOP stands for (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate;

[0224] BzCl represents benzoyl chloride;

[0225] mCPBA stands for meta-chloroperbenzoic acid;

[0226] Cbz represents benzyloxycarbonyl;

[0227] CDI stands for carbonyldiimidazole;

[0228] DAST stands for diethylaminosulfur trifluoride;

[0229] DBU stands for 1,8-diazabicycloundec-7-ene;

[0230] DCE stands for dichloroethane;

[0231] DCM stands for dichloromethane;

[0232] Dess-Martin periodinane represents 1,1,1-tris(acetoxy)-1,1-dihydro-1,2-benzidoyl-3-(1H)-one;

[0233] DIAD stands for diisopropyl azodicarboxylate;

[0234] DIBAL-H stands for diisobutylaluminum hydride;

[0235] DMAP stands for N,N-dimethylaminopyridine;

[0236] DME stands for 1,2-dimethoxyethane;

[0237] DMF stands for N,N-dimethylformamide;

[0238] DMSO stands for dimethyl sulfoxide;

[0239] DPPA stands for diphenylphosphoryl azide or diphenylphosphoryl azide;

[0240] Dppf stands for 1,1′-bis(diphenylphosphino)ferrocene;

[0241] EDCI or EDC stands for 1-(3-diethylaminopropyl)-3-ethylcarbodiimide hydrochloride;

[0242] EA or EtOAc represents ethyl acetate;

[0243] Ghosez reagent stands for 1-chloro-N,N,2-trimethyl-1-propenylamine;

[0244] HATU stands for O(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate;

[0245] HCl stands for hydrochloric acid;

[0246] Hunig's base represents diisopropylethylamine;

[0247] PyBOP stands for (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate;

[0248] LDA stands for lithium diisopropylamide;

[0249] Pd-C stands for palladium on carbon;

[0250] PE stands for petroleum ether;

[0251] Ph stands for phenyl;

[0252] RT stands for reverse transcription;

[0253] RT-PCR stands for reverse transcription-polymerase chain reaction;

[0254] TBME stands for tert-butyl methyl ether;

[0255] TEA stands for triethylamine;

[0256] Tf2O represents trifluoromethanesulfonic anhydride;

[0257] TFA stands for trifluoroacetic acid;

[0258] THF stands for tetrahydrofuran;

[0259] (TMS)2NH represents hexamethyldisilazane;

[0260] TBS stands for tert-butyldimethylsilyl;

[0261] TBDPS stands for tert-butyldiphenylsilane;

[0262] TMS stands for trimethylsilyl;

[0263] TPAP stands for tetrapropylammonium perruthenate;

[0264] TPP or PPh3 represents triphenylphosphine;

[0265] Ts or p-toluenesulfonyl represents p-CH3C6H4SO2-;

[0266] tBOC or Boc represents tert-butyloxycarbonyl; and

[0267] Palladium methanesulfonate represents 4,5-bisdiphenylphosphine-9,9-dimethyl-9H-xanthene.

[0268] Synthesis Method

[0269] The compounds and methods of the present invention will be better understood in conjunction with the following synthetic schemes, which illustrate methods for preparing the compounds of the present invention, which are intended to be illustrative only and do not limit the scope of the present invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art, and may be made without departing from the spirit of the present invention and the scope of the appended claims, including but not limited to changes and modifications related to the chemical structures, substituents, derivatives and / or methods of the present invention.

[0270] Scheme 1 illustrates a method for preparing compounds of formula 11 from compounds 1 and 2, wherein n = 1, 2 or 3; P is a hydroxy protecting group; Ar is E; and E is as described above. Alkylation of hydroxypyridine 1 with a hydroxy epoxide using Mitsunobu reaction conditions affords epoxide 4. Alternatively, the hydroxy epoxide is converted to 3 with a leaving group such as, but not limited to, tosyl and mesyl, followed by alkylation in the presence of a base such as, but not limited to, K2CO3 and Cs2CO3 to afford 4. Intramolecular epoxide ring opening mediated by a base such as, but not limited to, LDA yields compound 5. Hydroxyl compound 5 is protected with an appropriate protecting group such as, but not limited to, TBDPS and TBS to afford compound 6. Trifluoromethyl ketone 7 is obtained by iodomagnesium exchange of compound 6 followed by the addition of an ester such as, but not limited to, ethyl 2,2,2-trifluoroacetate. Trifluoromethyl ketone 7 is cross-coupled with various metal coupling agents 8, including but not limited to boric acid, boric acid esters, organotin reagents, organozinc reagents, organomagnesium reagents, organosilicon reagents, etc., catalyzed by appropriate catalysts such as Pd, Ni, Cu, etc., to give compound 9. Nitromethane is added to compound 9 in the presence of a base such as but not limited to K2CO3 and Cs2CO3 to give compound 10. Reduction of the nitro group with a reducing agent such as but not limited to zinc and acetic acid produces key intermediate 11.

[0271] Solution 1

[0272]

[0273] As shown in Scheme 2, Ar1 is A; Ar is E; R is R 11 ; n is 1, 2 or 3; A, E, R 11 As described above. Key intermediate 11 is coupled with various carboxylic acids to give amide 14. Amide 14 is then reacted with various electrophiles to produce various ethers, esters, and carbamates of formula 15. Amide 14 is also oxidized to aldehyde 16, followed by reductive amination to provide various amines 17. The -CH2OH hydroxyl group in amide 14 is converted to cyanomethyl 18 by activation followed by hydrogenation. Compound 14-1 is further converted to acetamide 19 in the presence of a catalyst, such as, but not limited to, Parkin's catalyst.

[0274] Solution 2

[0275]

[0276] As shown in Scheme 3, Ar1 is A; Ar is E; R is R 11 ; A, E, R 11 As described above, aldehyde 16 was converted to the benzyl-protected amine by reductive amination. Hydrogenolysis gave the free amine 20. Finally, displacement with various electrophiles gave the N-substituted compound 21.

[0277] Solution 3

[0278]

[0279] As shown in Scheme 4, wherein Ar1 is A; Ar is E; R' is -C1-C6 alkyl, -C3-C6 cycloalkyl, aryl or heteroaryl; n is 1, 2 or 3; A and E are as described above. After oxidation of aldehyde 16 to acid 22, it is further converted to amide 23 and sulfonamide 24 using common methods such as, but not limited to, HATU and DIPEA. From here, a variety of esters and amides are obtained.

[0280] Solution 4

[0281]

[0282] Scheme 5 shows another method for preparing compounds of formula 11, wherein Ar is E; P is a hydroxyl protecting group; n is 1, 2 or 3; E is as described above. Ketone 9 is converted to a compound of formula 26 by olefination. Alternatively, 26 can be obtained from the following sources: 1) 6 is cross-coupled with a metal coupling agent 6-1, which can be, but is not limited to, boric acid, boric acid ester, organotin reagent, organozinc reagent, organomagnesium reagent, organosilicon reagent, etc., catalyzed by a suitable catalyst such as Pd, Ni, Cu, etc., to obtain compound 25; 2) Compound 25 is converted to compound 26 as previously described in Scheme 1. With 26, a compound of formula 27 is prepared by dihydroxylation and then epoxide formation. Compound 27 is subjected to epoxide ring opening with an amine equivalent (such as, but not limited to, NH4OH and NH3) to obtain a compound of formula 11.

[0283] Solution 5

[0284]

[0285] Scheme 6 illustrates another method for preparing compounds of formula 23, wherein Ar1 is A; Ar is E; R' is -C1-C6 alkyl, -C3-C6 cycloalkyl, aryl or heteroaryl; n is 1, 2 or 3; A and E are as described above. Amine 11 is protected by a protecting group such as, but not limited to, Boc and Cbz. After deprotection of the hydroxy protecting group, oxidation is then performed to provide acid 30. Compound 30 is coupled with various amines to provide amide 31. Deprotection of the amine protecting group followed by amide formation provides compounds of formula 23.

[0286] Solution 6

[0287]

[0288] Scheme 7 illustrates an alternative route to the desired compounds. This route differs in that it starts with oxidation and amide coupling to install the amide 33 at the beginning of the synthesis. Sequential vinylation and arylation afford the bis-coupled product 34. Asymmetric dihydroxylation followed by activation and substitution affords the amino alcohol precursor. Finally, the amide is coupled with the corresponding aryl acid to yield the desired compound shown as compound 36.

[0289] Solution 7

[0290]

[0291] Example

[0292] The compounds and methods of the present invention will be better understood in conjunction with the following examples, which are intended to illustrate and not to limit the scope of the present invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art, and these changes and modifications may be made without departing from the spirit of the present invention and the scope of the appended claims, including but not limited to changes and modifications related to the chemical structures, substituents, derivatives, preparations and / or methods of the present invention.

[0293] Certain synthetic methods useful for preparing compounds of the present invention are disclosed in US2022 / 0356189, the entire contents of which are incorporated herein by reference.

[0294] Intermediate 1

[0295]

[0296] Intermediate 1 Steps a and b

[0297]

[0298] A solution of methyl 4-amino-3-methoxybenzoate (10 g, 55.19 mmol) in HCl (80 mL) was treated with methacrolein (9.67 g, 137.97 mmol) at 100 °C for 5 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The crude product was used directly in the next step without further purification. ESI-MS m / z: 217.95 [m+H] + .

[0299] A solution of the compound from step a (10 g, 46.03 mmol) in MeOH (100 mL) was treated with SOCl2 (10 mL, 137.86 mmol) at 0°C. The final reaction mixture was reacted at 80°C for 40 minutes. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The reaction was adjusted to pH = 8 with NaHCO3 at room temperature. The resulting mixture was extracted with EA. The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (1:1) to give the desired product (4.386 g, 41%) as a brown solid. ESI-MS m / z: 232.00 [m+H] + .

[0300] Intermediate 1 Steps c and d

[0301]

[0302] A mixture of DCM (20 mL) containing the methyl compound (3.9 g, 16.86 mmol) of step b and m-CPBA (8.73 g, 50.59 mmol) was stirred at room temperature overnight. The reaction was quenched with saturated sodium thiosulfate (aqueous solution) at room temperature. The resulting mixture was extracted with CH2Cl2. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with CH2Cl2 / MeOH (15:1) to give the desired product (2.6 g, 62%) as a yellow solid. ESI-MS m / z: 248.05 [m+H] + .

[0303] A solution of the compound from step c (2.6 g, 10.51 mmol) and POCl3 (20.96 g, 136.70 mmol) was stirred at 95 °C for 1 hour. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure and quenched with water / ice. The aqueous layer was extracted with EA. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (5:1) to give the desired product (1.5 g, 53%) as a yellow solid. ESI-MS m / z: 266.00 [m+H] + .

[0304] Intermediate 2

[0305]

[0306] Intermediate 2 Steps a and b

[0307]

[0308] A solution of (2Z)-2-bromobut-2-enal (5 g) and methyl 4-amino-3-hydroxybenzoate (10 g) in HCl (20 mL) and AcOH (30 mL) was stirred at 100°C under N2 for 1 hour. The resulting solution was concentrated to give the desired product (crude) as a brown solid. ESI-MS m / z: 282.00 [m+H] + .

[0309] A solution of MeOH (30 mL) containing the compound from step a (10 g, 35.45 mmol) and H2SO4 (10 mL) was stirred at 80 ° C for 2 hours. The mixture / residue was neutralized to pH 7-8 with NaHCO3. The resulting mixture was extracted with EA (x 3). The combined organic layers were washed with water and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA to give the desired product (1 g, 9.5%) as a yellow solid. ESI-MS m / z: 296.00 [m+H] + .

[0310] Intermediate 2 Steps c and d

[0311]

[0312] A solution of DMF (10 mL) containing the compound from step b (1 g, 3.38 mmol), sodium 2-chloro-2,2-difluoroacetate (1 g, 6.56 mmol) and Cs2CO3 (2.20 g, 6.75 mmol) was stirred at 80°C for 2 hours. The residue was purified by silica gel column chromatography eluting with PE / EA to give the desired product (800 mg, 68%) as a yellow solid. ESI-MS m / z: 346.00 [m+H] + .

[0313] A solution / mixture of MeOH (10 mL) and H2O (10 ml) containing the compound of step c (800 mg, 2.31 mmol) and LiOH (553 mg, 23.11 mmol) was stirred at room temperature under N2 for 6 hours. The crude product was recrystallized from MeOH / H2O to give the desired product (500 mg, 65%) as a yellow solid. ESI-MS m / z: 332.00 [m+H] + .

[0314] Intermediate 3

[0315]

[0316] Intermediate 3 Step a

[0317]

[0318] A 100 mL round-bottom flask was charged with 2-cyclopropyl-7-methoxy-2H-indazole-5-carboxylic acid (500 mg, 2.153 mmol), toluene (4.98 mL) and methanol (2.99 mL), then cooled to 0 ° C in an ice bath. TMS diazomethane (2.8 mL, 5.60 mmol) was added dropwise manually over 10 minutes. The mixture was stirred for 1 hour, and then the diazo reagent was quenched by adding AcOH until the yellow color of the diazo reagent faded. The mixture was then concentrated and sent directly to the next step ESI-MS m / z: 247.213 [m+H] + .

[0319] Intermediate 3 Steps b

[0320]

[0321] Into a 100mL round-bottom flask with a stirring bar, 2-cyclopropyl-7-hydroxy-2H-indazole-5-carboxylic acid methyl ester (529mg, 2.15mmol), tetrabutylammonium iodide (1.032g, 2.80mmol) and CH2Cl2 (21.5ml) were loaded. The mixture was cooled to -78°C and trichloroborane (5.38ml, 5.38mmol) was added dropwise within 10 minutes. The mixture was stirred at -78°C for 1 hour, then stirred in an ice bath and warmed to 0°C for 1 hour. The mixture was then diluted with water (50ml) and ethyl acetate (50ml), the layers were separated, and the aqueous layer was washed with EtOAc (4x 50ml). The combined organic matter was washed with brine, dried with Na2SO4, filtered and concentrated to obtain a red oil, which was purified by silica gel chromatography to obtain the desired white solid product (489mg, 98% yield). ESI-MS m / z:233.201[m+H] + .

[0322] Intermediate 3 Step c

[0323]

[0324] A 100 mL round-bottom flask was charged with 2-cyclopropyl-7-hydroxy-2H-indazole-5-carboxylic acid (0.499 g, 2.15 mmol), cesium carbonate (1.401 g, 4.30 mmol), DMF (21.50 ml), and then sodium chlorodifluoroacetate (0.656 g, 4.30 mm). The mixture was heated to 105 ° C for 6 hours, then cooled to room temperature and purified by silica gel chromatography (containing 0 to 30% EtOAc in cyclohexane) to give the desired white solid compound (176.4 mg, 29% yield). ESI-MS m / z: 283.160 [m+H] + .

[0325] Intermediate 3 Step d

[0326]

[0327] 2-cyclopropyl-7-(difluoromethoxy)-2H-indazole-5-carboxylic acid methyl ester (174mg, 0.616mmol) was loaded into a 20mL vial and dissolved in MeOH (2.055ml), THF (2.055ml) and water (2.055mL). Lithium hydroxide (78mg, 1.849mmol) was freshly crushed and added. The reaction mixture was stirred for 2 hours. The reaction mixture was then carefully acidified by adding 1MHCl. The mixture was diluted with water and EtOAc. The aqueous layer was washed with EtOAc (3x 5mL), the combined organic layer was washed with brine, and then concentrated to obtain the desired product as a white solid (147.6mg, 89% yield). ESI-MS m / z:269.145[m+H] + .

[0328] Intermediate 4

[0329]

[0330] Intermediate 4 Step a

[0331]

[0332] 3-Chloro-8-methoxy-2-methylquinoline-6-carboxylic acid (200 mg, 0.795 mmol), toluene (1.84 mL) and methanol (1.104 mL) were placed in a 20 mL vial and then cooled to 0 °C in an ice bath. TMS-diazomethane (1.0 mL, 2.07 mmol) was added dropwise manually over 10 minutes. The mixture was stirred for 1 hour, then AcOH was added to quench the diazo reagent until the yellow color of the diazo reagent disappeared, and the mixture was concentrated to give the desired product (48 mg, 95%) as a yellow solid. ESI-MS m / z: 266.091 [m+H] + .

[0333] Intermediate Step 4b

[0334]

[0335] An 8 mL vial was charged with 3-chloro-8-methoxy-2-methylquinoline-6-carboxylic acid methyl ester (0.211 g, 0.795 mmol), then D2O (1.125) was added and the mixture was heated to 100°C for 18 hours. The mixture was then cooled to room temperature and diluted with EtOAc. The phases were separated and the organic layer was washed with 3 x 1 mL NaHCO3. The organic layer was wiped dry with Na2SO4, filtered, concentrated, and purified by combiflash to give the desired product as a yellow solid (105 mg, 34% yield) ESI-MS m / z: 269.156 / 271.135 [m+H] + .

[0336] Intermediate Step 4c

[0337]

[0338] A 20 mL vial was charged with methyl 3-chloro-8-methoxy-2-(methyl-d3)quinoline-6-carboxylate (100 mg, 0.372 mmol), dissolved in water (1.240 ml), THF (1.240 mL), methanol (1.240 mml), then freshly crushed lithium hydroxide (26.7 mg, 1.116 mmol) was added. The mixture was stirred for 1 hour, then diluted with DCM containing 10% MeOH, then carefully acidified by adding 1 M HCl, the phases separated, and the aqueous layer washed with DCM containing 10% MeOH. The combined organic phases were concentrated to give the desired product as a white solid (87 mg, 92% yield) ESI-MS m / z: 255.163 / 257.175 [m+H] + .

[0339] Intermediate 5

[0340]

[0341] An 8 mL vial was charged with 3-bromo-8-methoxy-2-methylquinoline-6-carboxylic acid suspended in 0.7 mL D2O and then heated to 100°C for 18 hours. The crude product was lyophilized and used without further purification (48 mg, 95%) as a yellow solid. ESI-MS m / z: 299.033 / 300.961 [m+H] + .

[0342] Intermediate 6

[0343]

[0344] Intermediate 6 Step a

[0345]

[0346] In a drying vial with a stirring bar, methyl 2-chloro-8-methoxy-3-methylquinoline-6-carboxylate (250 mg, 0.941 mmol) was dissolved in DMF (1.9 mL) and anhydrous Me4NF (175 mg, 2.0 equiv) was added. The vial was sealed and heated to 80 ° C overnight. The reaction mixture was then cooled to room temperature, diluted with DCM, washed twice with water and once with brine. The organic phase was then dried over sodium sulfate, filtered and concentrated. The crude residue was purified by automatic column chromatography (silica gel) to give a white solid fluoride, which was contaminated by some chloride starting material (48 mg, 20%). ESI-MS m / z: 250.134 [m+H] + .

[0347] Intermediate 6 Step b

[0348]

[0349] In a 1dram bottle, methyl 2-fluoro-8-methoxy-3-methylquinoline-6-carboxylate (48 mg, 0.18 mmol 1.0 equiv) was dissolved in THF (0.3 mL) and water (0.15 mL), and the mixture was cooled to 0 ° C. LiOH (8.7 mg, 0.36 mmol, 2.0 equiv) was added, the mixture was stirred for 15 minutes, and then the cooling bath was removed. After 2 hours at room temperature, the mixture was acidified to pH about 5 with AcOH, and the mixture was directly concentrated. The crude residue was purified by preparative HPLC (ACN / H2O, 20-90%, 25 minutes) to give the product carboxylic acid (13 mg, 31%). ESI-MS m / z: 236.16 [m+H] + .

[0350] Intermediate 7

[0351]

[0352] Intermediate 7 Step a

[0353]

[0354] The above compound was prepared from methyl 3-bromo-2-chloro-8-methoxyquinoline-6-carboxylate according to the following procedure: HCl (4M in 1,4-dioxane, 0.34 mL, 1.34 mmol / 1.0 eq.) was added to a suspension of methyl 3-bromo-2-fluoro-8-methoxyquinoline-6-carboxylate (443 mg, 1.34 mmol, 1.0 eq.) in DCM (5 mL), and the mixture was stirred at room temperature for 5 minutes. The solvent was then removed in vacuo, and the residue was suspended in MeCN (6.7 mL). NaI (1.0 g, 6.7 mmol, 5.0 eq.) was then added, the vial was sealed, and heated to 80 °C for 3 hours. The mixture was cooled to room temperature, and the solvent was removed in vacuo. The residue was redissolved in DCM, and the organic solution was then washed with 10% aqueous K2CO3 and water, dried over sodium sulfate, filtered, and concentrated. The crude residue was purified by automated column chromatography (silica gel) to give a white solid iodide (434 mg, 77%). ESI-MS m / z: 423.93 [m+H] + .

[0355] Intermediate Step 7b

[0356]

[0357] In a drying vial with a stirring rod, 3-bromo-2-iodo-8-methoxyquinoline-6-carboxylic acid methyl ester (75 mg, 0.18 mmol, 1.0 equiv) was suspended in THF (0.5 mL) and Et2O (0.5 mL), and the mixture was cooled to 0 ° C. Isopropylmagnesium chloride (0.11 mL, 0.22 mmol, 1.2 equiv) was added, the reaction mixture was stirred at this temperature for 1 hour, and then the reaction was quenched by the dropwise addition of CD3OD (0.6 mL, 14.8 mmol, 15 equiv). The mixture was stirred at room temperature for another 15 minutes and then poured into a saturated aqueous ammonium chloride solution. The quenched reaction mixture was diluted with water and DCM, the layers were separated, and the aqueous layer was extracted 3 times with DCM. The combined organic extracts were dried over sodium sulfate, filtered and concentrated. The crude material was purified by automated column chromatography (silica gel) to give the deuterated product methyl-3-bromo-8-methoxyquinoline-6-carboxylic acid-2-D (9.0 mg, 17%) as a white solid. ESI-MS m / z: 297.06 [m+H] + .

[0358] Intermediate 7 Step c

[0359]

[0360] In a 1dram bottle with a stir bar, methyl-3-bromo-8-methoxyquinoline-6-carboxylate-2-D (9.0 mg, 1.0 eq., 0.032 mmol) as a white solid was dissolved in THF (0.1 mL), MeOH (0.1 mL) and water (0.1 mL). LiOH (7 mg, 0.30 mmol, 10 eq.) was added and the reaction mixture was stirred at room temperature overnight. The mixture was then acidified to pH about 2 with 2M HCl and concentrated directly to give the carboxylic acid, which was used without further purification (8.0 mg, 93%). ESI-MS m / z: 283.05 [m+H] + .

[0361] Intermediate 8

[0362]

[0363] The above compound was prepared from 8-methoxy-2-methyl-3-(methyl-d3)quinoline-6-carboxylic acid in a similar manner to step X of Example X to give the hexadeuterated compound as an orange solid (27 mg, 96%). ESI-MS m / z: 238.16 [m+H] + .

[0364] Intermediate 9

[0365]

[0366] Intermediate 9 Step a

[0367]

[0368] Under nitrogen, a mixture of (R)-7-bromo-5-iodo-3-methyl-2,3-dihydrofuran[2,3-c]pyridine-3-carboxamide (2 g, 5.22 mmol), 2-(1-(4-fluorophenyl)vinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.57 g, 6.32 mmol), Pd(dppf)Cl2 (0.382 g, 0.52 mmol), K2CO3 (2.17 g, 15.67 mmol) in dioxane (21 mL) and H2O (5 mL) was stirred at 90 °C for 2 hours. The reaction was monitored by LCMS. The resulting mixture was filtered and the filter cake was washed with EA. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with a hexane solution containing 50% ethyl acetate) to give the desired product (1.97 g, 75 wt %, 75%) as a red oil. ESI-MS m / z: 377.06 [m+H] + .

[0369] Intermediate Step 9b

[0370]

[0371] A mixture of the compound from step a (1 g, 2.25 mmol), 2-(2,3-dichloro-4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.82 g, 2.82 mmol), Pd(dppf)Cl2 (0.165 g, 0.225 mmol) and K2CO3 (0.70 g, 5.07 mmol) in dioxane (9 mL) and H2O (2.25 mL) was stirred at 90°C for 2 hours under nitrogen. The reaction was cooled to room temperature and the mixture was diluted with EtOAc and saturated ammonium chloride. The aqueous phases were extracted with EtOAc, combined, dried over sodium sulfate, filtered and concentrated. The crude material was purified by automated column chromatography (silica gel, 0-100% ethyl acetate in cyclohexane) to give the desired product (1.50 g, 80% wt, 87%) as a red oil. ESI-MS m / z:461.12[m+H] + .

[0372] Intermediate 9 Step c

[0373]

[0374] The t-BuOH (17 mL) solution containing the compound (1.5 g, 3.38 mmol) of step b and H2O (17 mL) were cooled to 0 ° C. Methanesulfonamide (0.64 g, 6.77 mmol) and AD-mix-β (13.18 g, 16.92 mmol) were added, and the reactants were stirred at room temperature overnight and monitored by LCMS. The reaction was cooled to room temperature, the mixture was diluted with EtOAc, and quenched with sodium sulfite. The aqueous phase was extracted with EtOAc, combined, dried over sodium sulfate, filtered and concentrated. The crude material was purified by automatic column chromatography (silica gel, cyclohexane solution containing 0-100% ethyl acetate) to obtain the desired product (1.0 g, 60%) as a yellow solid. ESI-MS m / z: 495.09 [m + H] + .

[0375] Intermediate 9 Step d

[0376]

[0377] A solution of DCM (11 mL) containing the compound from step c (1.0 g, 2.01 mmol) was cooled to 0 °C. TsCl (0.654 g, 3.43 mmol), TEA (0.84 mL, 6.06 mmol) and DMAP (0.24 g, 2.01 mmol) were added and the reaction was stirred at room temperature for 90 minutes. The crude reaction was concentrated and the material was purified by automated column chromatography (silica, cyclohexane solution containing 0-100% ethyl acetate) to give the desired product (1.6 g, 70% wt, 85%) as a yellow solid. ESI-MS m / z: 649.10 [m+H] + .

[0378] Intermediate 9 Step e

[0379]

[0380] To a 250 mL round bottom flask containing step d (1.6 g, 3.04 mmol) was added NH3 (55 mL, 156 equiv., 7 N in methanol) at room temperature. The resulting mixture was stirred at room temperature for 20 hours and monitored by LCMS. The solvent was removed and the crude mixture was dissolved in EtOAc. The organics were washed 3 times with saturated sodium bicarbonate and concentrated. The crude material was triturated with DCM to give the desired product (425 mg, 35%) as an off-white solid. ESI-MS m / z: 494.11 [m+H] + .

[0381] Intermediate 10

[0382]

[0383] The above compound was prepared in a similar manner to Intermediate 9. ESI-MS m / z: 478.17 [m+H] + .

[0384] Intermediate 11

[0385]

[0386] The above compound was prepared in a similar manner to Intermediate 9. ESI-MS m / z: 470.17 [m+H] + .

[0387] Intermediate 12

[0388]

[0389] The above compound was prepared by a similar method as Intermediate 9 to give the amino alcohol. (133 mg, quantitative yield). ESI-MS m / z: 440.13 [m+H]+ .

[0390] Intermediate 13

[0391]

[0392] The above compound was prepared by a similar method as Intermediate 9 to give the amino alcohol. (158 mg, quantitative yield). ESI-MS m / z: 522.18 [m+H] + .

[0393] Intermediate 14

[0394]

[0395] The above compound was prepared by a similar method as Intermediate 9 to give the amino alcohol (170 mg, quantitative yield). ESI-MS m / z: 528.27 [m+H] + .

[0396] Example 1

[0397]

[0398] (R)-5-((S)-2-amino-1-cyclopropyl-1-hydroxyethyl)-7-(5-chloro-2,4-difluorophenyl)-3-methyl-2,3-dihydrofuro[2,3-c]pyridine-3-carboxamide (15 mg, 0.035 mmol) and 8-methoxy-3-(trifluoromethyl)quinoline-6-carboxylic acid (10.56 mg, 0.039 mmol) were added to a 20 mL vial equipped with a stirring bar. The solid was dissolved in DMF (0.20 mL) and DIPEA (18.54 μl, 0.106 mmol) was added. The vial was cooled to 0 ° C and PyBOP (22.10 mg, 0.042 mmol) was added. The reactants were stirred for 10 minutes, heated to room temperature, and monitored by LCMS (1 hour). After the reaction was complete, the reactants were diluted with EtOAc and quenched with saturated ammonium chloride. The aqueous phase was extracted with EtOAc, collected with a phase separator cartridge and concentrated. The crude material was purified by preparative HPLC (ACN / H2O, 20-90%, 25 min), fractions were concentrated with a Biotage V10 evaporator, and the material was lyophilized to give a fluffy white solid (13 mg, 53%) as the desired product. ESI-MS m / z: 677.20 [m+H] + .

[0399] Table 1 below contains examples prepared in a manner similar to Example 1. Most compounds were purified by preparative HPLC (ACN / H2O, 20-90%, 25 minutes) and some compounds were purified by automated column chromatography (silica gel). Aryl acid coupling agents were prepared according to Intermediates 1-14, or by similar methods with slight modifications, also according to the methods in U.S. Patent Application No. 17 / 679,746.

[0400] Table 1

[0401]

[0402]

[0403]

[0404]

[0405]

[0406] Intermediate 15

[0407]

[0408] Intermediate 15 Steps a and b

[0409]

[0410] 2-(1-cyclopropylvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (6.33 g, 32.6 mmol) was added to a 250 mL round bottom flask and the oil was dissolved in 1,4-dioxane (104 mL). (R)-7-bromo-5-iodo-3-methyl-2,3-dihydrofurano[2,3-c]pyridine-3-carboxamide (10 g, 26.1 mmol) was added followed by PdCl2(dppf) (1.91 g, 2.61 mmol) and water (26.1 mL). Potassium carbonate (10.83 g, 78 mmol) was then added and the round bottom flask was equipped with a condenser and a N2 bubbling system and the reaction was heated to 90°C for 2 hours. The reaction was cooled to room temperature and the mixture was diluted with EtOAc and saturated ammonium chloride. The aqueous phase was extracted with EtOAc, combined, dried over sodium sulfate, filtered and concentrated. The crude material was purified by automated column chromatography (silica gel, cyclohexane solution containing 0-100% ethyl acetate) to give the desired product (10.38 g, 70% wt, 86%) as a red oil. ESI-MS m / z: 323.20 [m+H] + .

[0411] A solution of (R)-7-bromo-5-(1-cyclopropylvinyl)-3-methyl-2,3-dihydrofuro[2,3-c]pyridine-3-carboxamide (3.90 g, 12.07 mmol), K2OsO4·2H2O (0.19 g, 0.52 mmol) and NMO (3.96 g, 33.79 mmol) in THF (10 mL), H2O (10 mL) and acetone (10 mL) was stirred at room temperature overnight. The reaction was quenched by the addition of Na2S2O3 (20 mL) at room temperature. The aqueous layer was extracted with EA (3×200 mL). The residue was purified by silica gel column chromatography eluting with PE / EA to give the desired product (2.60 g) as a yellow solid. ESI-MS m / z: 356.90 [m+H] + .

[0412] Intermediate 15 Steps c and d

[0413]

[0414] A solution of DCM (50 mL) containing the compound from step b (2.80 g, 7.84 mmol), TsCl (1.79 g, 9.41 mmol), DMAP (48 mg, 0.39 mmol) and TEA (2.38 g, 23.52 mmol) was stirred at 0 ° C for 2 hours. The mixture was acidified to pH 7-8 with HCl. The resulting mixture was extracted with EA (3 x 200 mL). The combined organic layers were washed with water (3 x 200 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA to give the desired product (2 g, 50%) as a yellow solid. ESI-MS m / z: 510.90 [m+H] + .

[0415] To a stirred solution of MeOH (200 mL) containing NH3 (g) was added a solution of MeOH (3 mL) containing the compound of step c (3 g, 5.87 mmol) at room temperature. The mixture was stirred at room temperature for 20 hours. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in EtOAc and washed with saturated NaHCO3 aqueous solution. The desired product (2 g, 96%) was concentrated to give a white solid. ESI-MS m / z: 355.95 [m+H] + .

[0416] Intermediate 15 Step e

[0417]

[0418] A solution of the compound of step d (700 mg, 1.97 mmol) in DMF (5 mL) was treated with 2-cyclopropyl-7-methoxy-2H-indazole-5-carboxylic acid (391 mg, 1.68 mmol), PyBOP (877 mg, 1.69 mmol) and DIPEA (363 mg, 2.81 mmol) at room temperature for 1 hour. The reaction was monitored by LCMS. The residue was purified by reverse phase flash chromatography to give the title compound (1 g, 86%) as a white solid. ESI-MS m / z: 570.30 [m+H] + .

[0419] Intermediate 16

[0420]

[0421] The above compound was prepared in a similar manner to Intermediate 15 and purified by automated column chromatography (silica gel) to give pyridyl bromide (812 mg, 85%) as a white solid. ESI-MS m / z: 610.02 [m+H] + .

[0422] Embodiment 47

[0423]

[0424] To a 2 dram bottle equipped with a stir bar was added Pd(PPh3)Cl2 (7.38 mg, 10.52 μmol), sodium carbonate (22.30 mg, 0.210 mmol), (5-chloro-2-fluoro-3-methylphenyl)boronic acid (19.82 mg, 0.105 mmol) and (3R)-7-bromo-5-(1-cyclopropyl-2-(2-cyclopropyl-7-methoxy-2H-indazole-5-carboxamido)-1-hydroxyethyl)-3-methyl-2,3-dihydrofuro[2,3-c]pyridine-3-carboxamide (40 mg, 0.070 mmol). The vial was bubbled with N2 and the solid was dissolved in 1,4-dioxane (0.28 mL) and water (0.07 mL). The reaction was heated to 90°C and monitored by LCMS (1 hour). The reaction was cooled to room temperature, filtered on a silica gel pad with EtOAc and concentrated. The crude material was purified by preparative HPLC (ACN / H2O, 20-90%, 25 minutes) and the diastereomers were separated. The fractions were concentrated using a Biotage V10 evaporator and the material was lyophilized to give a fluffy white solid (6 mg, 13%) as the desired product. ESI-MS m / z: 634.31 [m+H] + .

[0425] Table 2 below contains examples prepared in a manner analogous to Example 47 using intermediates 15-16 or their analogs. More palladium and boric acid may be added if needed to drive conversion. Most compounds were purified and diastereomers separated by preparative HPLC (ACN / H2O, 20-90%, 25 minutes). If reported as a mixture of diastereomers, they may not be separated by HPLC.

[0426] Table 2

[0427]

[0428]

[0429]

[0430] Intermediate 17

[0431]

[0432] Intermediate 17 Step a

[0433]

[0434] A solution of CHCl3 (10 mL) containing 2,2-difluoro-1,3-benzodioxol-4-amine (5.00 g, 28.88 mmol) was treated with NBS (5.66 g, 31.80 mmol) at room temperature for 1 hour. The reaction was monitored by LCMS. The resulting mixture was extracted with CH2Cl2. The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (2:1) to give a yellow oily compound (5.66 g, 78%). ESI-MS m / z: 253.80 [m+H] + .

[0435] Intermediate 17 Step b

[0436]

[0437] Under nitrogen, a MeCN (10 mL) solution containing CuCl (3.20 g, 23.81 mmol) was treated with t-BuNO (3.07 g, 29.76 mmol) at 55 ° C for 10 minutes, and then the compound (3 g, 11.90 mmol) in step a was added dropwise at 55 ° C. The final reaction mixture was heated at 55 ° C for 30 minutes. The reaction was monitored by LCMS. The mixture was acidified to pH 3 with concentrated hydrochloric acid. HCl. The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA 10:1) to obtain the title compound (396.6 mg, 7%) as a white oil. 1 HNMR (400MHz, CDCl3) δ6.92 (d, J=8.6Hz, 1H), 7.40 (d, J=8.6Hz, 1H).

[0438] Intermediate 17 Step c

[0439]

[0440] Under nitrogen, a solution of THF (10 mL) containing the compound of step b (2.87 g, 10.57 mmol) was treated with iPrMgCl-LiCl (1.3 M in THF solution, 8 mL, 10.57 mmol) at -20 ° C for 30 minutes, and then 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.36 g, 12.69 mmol) was added dropwise at -20 ° C. The final reaction mixture was reacted at room temperature for 1 hour. The reaction was monitored by H-NMR. The resulting mixture was extracted with CH2Cl2. The combined organic layer was dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (4:1) to give the title compound (2.25 g, 67%) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ1.36 (s, 12H), 6.96 (d, J=8.1Hz, 1H), 7.52 (d, J=8.1Hz, 1H).

[0441] Intermediate 18

[0442]

[0443] Intermediate 18 Steps a and b

[0444]

[0445] A DMF (70 mL) solution containing 5-bromo-1-fluoro-3-methoxy-2-nitrobenzene (10 g, 40.00 mmol) was treated with Cs2CO3 (13.03 g, 40.000 mmol) and benzyl alcohol (8.65 g, 80.00 mmol) at 70 ° C overnight. The reaction was monitored by LCMS. The resulting mixture was filtered and the filter cake was washed with ethyl acetate. The filtrate was extracted with EtOAc. The combined organic layer was washed with brine and concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (9:1) to give a yellow oily crude compound (15 g).

[0446] A solution of the compound of step a (15 g, 44.65 mmol) in ethanol (100 mL) was treated with a solution of NH4Cl (47.77 g, 893.08 mmol) in H2O (100 mL) and Fe (7.48 g, 133.96 mmol) at 80 ° C for 30 minutes. The reaction was monitored by LCMS. The resulting mixture was filtered and the filter cake was washed with ethyl acetate. The filtrate was concentrated under reduced pressure. The resulting mixture was extracted with EtOAc. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (9: 1) to give the desired compound (8.9 g, 65%) as a brown oil. ESI-MS m / z: 307.95 [m+H] + .

[0447] Intermediate 18 Steps c and d

[0448]

[0449] A solution of THF (150 mL) containing the compound of step b (6 g, 19.47 mmol) was treated with Ac2O (4.97 g, 48.67 mmol) and Et3N (9.85 g, 97.35 mmol) at 0°C to room temperature overnight. The reaction was monitored by LCMS. The resulting mixture was extracted with EtOAc. The combined organic layers were concentrated under reduced pressure. The residue was purified by trituration with PE to give the desired compound (6.24 g, 92%) as a white solid. ESI-MS m / z: 350.05 [m+H] + .

[0450] A solution of CH2Cl2 (200 mL) containing the compound from step c (6.21 g, 17.73 mmol) was treated with boron trichloride (88.66 mL, 88.66 mmol) at 0°C. The reaction was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc. The combined organic layers were concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to give the desired compound (1.5 g, 33%) as a white solid. ESI-MS m / z: 259.95 [m+H] + .

[0451] Intermediate 18 Steps e, f and g

[0452]

[0453] A solution of CHCl3 (10 mL) containing the compound of step d (1.5 g, 5.77 mmol) was treated with POCl3 (1.33 g, 8.65 mmol) at 65 ° C overnight. The reaction was monitored by LCMS. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (5:1) to give the desired compound (980 mg, 70%) as a white solid. ESI-MS m / z: 241.95 [m+H] + .

[0454] DPPP (852 mg, 2.07 mmol), Pd(OAc)2 (232 mg, 1.03 mmol) were added to a solution of DMF (6 mL), MeOH (6 mL) and TEA (1.5 mL) containing the compound from step e (1.25 g, 5.16 mmol) in a pressure tank. The mixture was bubbled with carbon monoxide and then pressurized to 20 atm with carbon monoxide at 100 ° C overnight. The reaction was monitored by LCMS. The resulting mixture was extracted with EtOAc. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (3: 1) to give the title compound (730 mg, 64%) as a white solid. ESI-MS m / z: 222.10 [m+H] + .

[0455] A solution of THF (8 mL) and MeOH (2 mL) containing step f (400 mg, 1.81 mmol) was treated with a solution of H2O (2 ml) containing LiOH (433 mg, 18.08 mmol) at room temperature for 2 hours. The reaction was monitored by LCMS. The product was precipitated by the addition of HCl. The precipitated solid was collected by filtration and washed with water to give the title compound (249.8 mg, 66%) as a white solid. ESI-MS m / z: 208.10 [m+H] + .

[0456] Intermediate 19

[0457]

[0458] Intermediate 19 Step a

[0459]

[0460] A solution of the compound of step b of intermediate 18 (2.7 g, 8.76 mmol) in dioxane (20 mL) was treated with cyclopropanecarbonyl chloride (1.1 g, 10.52 mmol) at 100 °C for 5 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by trituration with diethyl ether. The precipitated solid was collected by filtration and washed with diethyl ether to give the desired compound (2.78 g, 88%) as a white solid. ESI-MS m / z: 376.05 [m+H] + .

[0461] Intermediate 19 Steps b, c, d and e

[0462]

[0463] The following compound was prepared in a similar sequence to Intermediate 18 above and purified by reverse phase flash chromatography to give the title compound as a white solid (416.4 mg, 58%). ESI-MS m / z: 234.10 [m+H] + .

[0464] Intermediate 20

[0465]

[0466] Intermediate 20 Step a

[0467]

[0468] A solution of methanol (50 mL) containing methyl 3,5-difluoro-4-nitrobenzoate (3 g, 13.82 mmol) was treated with KOH (853 mg, 15.20 mmol) at 70 ° C for 3 hours. The reaction was monitored by TLC. The resulting mixture was extracted with EtOAc. The combined organic layers were concentrated under reduced pressure. The crude product was used directly in the next step without further purification.

[0469] Intermediate 20 Steps b and c

[0470]

[0471] To a flask containing the compound from step a (2.50 g, 10.91 mmol) and 2-methylsulfonylethanol (1.42 g, 11.45 mmol) was added DMSO (50 mL) and t-BuOK (1.29 g, 11.45 mol) under N2. The reaction was stirred for 20 hours under N2 at room temperature. Another portion of 2-methylsulfonylethanol (1.42 g, 11.45 mmol) and t-BuOK (1.29 g, 11.45 mmol) were added and the reaction was stirred for another 3.5 hours. The reaction was monitored by LCMS. The mixture / residue was acidified to pH <1 with concentrated hydrochloric acid. The resulting mixture was extracted with EtOAc. The aqueous layer was concentrated under reduced pressure. The resulting mixture was used directly in the next step without further purification. ESI-MS m / z: 211.95 [mH] - .

[0472] A solution of MeOH (10 mL) containing the compound from step b (400 mg, 1.88 mmol) was treated with H2SO4 (1 mL) at 70°C for 2 hours. The reaction was monitored by LCMS. The resulting mixture was extracted with EtOAc. The combined organic layers were concentrated under reduced pressure. The crude product was used directly in the next step without further purification. ESI-MS m / z: 225.95 [mH] - .

[0473] Intermediate 20 Step d

[0474]

[0475] At 80 ° C, a solution of methyl compound (1.4 g, 6.16 mmol) in ethanol (50 mL) containing step c was treated with NH4Cl (6.5 g, 123.26 mmol) in H2O (50 mL) and Fe (1.03 g, 18.49 mmol) for 30 minutes. The reaction was monitored by LCMS. The resulting mixture was filtered and the filter cake was washed with EtOAc. The filtrate was concentrated under reduced pressure. The resulting mixture was extracted with EtOAc. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (1: 1) to give the title compound (428 mg, 35%) as a white solid. ESI-MS m / z: 198.10 [m+H] + . 1 HHMR (400MHz, DMSO-d6) δ3.75 (s, 3H), 3.79 (s, 1H), 7.00 (d, J = 1.8Hz, 1H), 7.09 (dd, J = 1.8, 0.8Hz, 1H), 9.45 (s, 1H).

[0476] Intermediate 21

[0477]

[0478] Intermediate 21 Steps a and b

[0479]

[0480] A solution of 5-bromo-1,3-difluoro-2-nitrobenzene (5 g, 21.00 mmol), cyclopropanol (1.22 g, 21.0 mmol) and Cs2CO3 (13.69 g, 42.00 mmol) in DMF (50 mL) was stirred overnight at 60° C. The resulting mixture was extracted, concentrated and purified by silica gel column chromatography to give the desired compound (4.4 g, 75%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) δ0.70-0.94 (m, 4H), 4.17-4.26 (m, 1H), 7.60-7.65 (m, 1H), 7.68 (t, J=1.8Hz, 1H).

[0481] A solution of DMSO (40 mL) containing the compound from step a (4 g, 14.50 mmol), 2-methylsulfonylethanol (1.89 g, 15.20 mmol) and t-BuOK (1.71 g, 15.20 mol) was stirred at room temperature overnight. 2-Methylsulfonylethanol (1.89 g, 15.20 mmol) and t-BuOK (1.71 g, 15.20 mmmol) were added and the mixture was stirred at room temperature for another 3 hours. The mixture was acidified, extracted, concentrated and purified by silica gel column chromatography to give the desired compound (2.6 g, 65%) as a yellow solid. ESI-MS m / z: 271.95 [mH] - .

[0482] Intermediate 21 Steps c and d

[0483]

[0484] A solution of ethanol (20 mL) containing the compound from step b (2.6 g, 9.50 mmol), NH4Cl (5.07 g, 95.00 mmol), Fe (5.30 g, 95.00 mmol), H2O (10 mL) was stirred at 80°C for 30 minutes. The resulting mixture was filtered, concentrated and purified by silica gel column chromatography to give the desired compound (2 g, 86%) as a brown solid. ESI-MS m / z: 245.90 [m+H] + .

[0485] A solution of THF (20 mL) containing the compound from step c (1.5 g, 6.10 mmol), cyclopropanecarbonyl chloride (0.58 g, 5.50 mmol) and TEA (0.93 g, 9.20 mmol) was stirred at 0°C for 1 hour. The resulting mixture was extracted, concentrated and purified by silica gel column chromatography to give the desired compound (1.2 g, 62%) as a brown solid. ESI-MS m / z: 313.90 [m+H] + .

[0486] Intermediate 21 Steps e and f

[0487]

[0488] A solution of CHCl3 (20 mL) containing the compound of step d (1.2 g, 3.80 mmol) and POCl3 (884 mg, 5.70 mmol) was stirred at 65°C for 24 hours. The resulting mixture was extracted, concentrated and purified by silica gel column chromatography to obtain the desired compound (1 g, 88%) as a yellow oil. ESI-MS m / z: 293.90 [m+H] + .

[0489] A solution of the compound from step e (660 mg, 2.20 mmol), Ac2O (458 mg, 4.49 mmol), DIPEA (254 mg, 1.97 mmol), XantPhos (57 mg, 0.10 mmol), oxalic acid (404 mg, 4.49 mmol) and Pd(AcO)2 (50 mg, 0.22 mmol) in DMF (5 mL) was stirred at 100°C for 6 hours under nitrogen. The resulting mixture was filtered and purified by reverse phase flash chromatography to give the desired compound (235.5 mg, 40%) as a white solid. ESI-MS m / z: 260.10 [m+H] + . 1 H NMR(400MHz, DMSO-d6)δ0.72-0.79(m,2H),0.79-0.91(m,2H),1.11-1.27(m,4H),2.23-2. 34(m,1H),4.06-4.15(m,1H),7.73(d,J=1.3Hz,1H),7.77(d,J=1.3Hz,1H),13.12(s,1H).

[0490] Intermediate 22

[0491]

[0492] Intermediate 22 Steps a and b

[0493]

[0494] Under nitrogen, a solution of 5-bromo-1-chloro-3-fluoro-2-nitrobenzene (4 g, 15.70 mmol), 2-methylsulfonylethanol (2.05 g, 16.50 mmol) and t-BuOK (1.85 g, 16.5% mmol) in DMSO (40 mL) was stirred at room temperature overnight. 2-Methylsulfonylethanol (2.05 g, 16.50 mmol) and t-BuOK (1.85 g, 16.5 mmol) were added and the resulting mixture was stirred at room temperature for another 3 hours. The resulting mixture was extracted, concentrated and purified by silica gel column chromatography to give the desired compound (2.2 g, 55%) as a yellow solid. ESI-MS m / z: 251.85 [mH] - .

[0495] A solution of the compound from step a (2.2 g, 8.70 mmol), Fe (4.87 g, 87.10 mmol), NH4Cl (4.66 g, 87.1 mmol), H2O (10 mL) in ethanol (20 mL) was stirred at 80°C for 30 minutes. The resulting mixture was concentrated and purified by silica gel column chromatography to give the desired compound (1.2 g, 61%) as a brown solid. ESI-MS m / z: 223.85 [m+H] + .

[0496] Intermediate 22 Steps c, d and e

[0497]

[0498] A solution of THF (10 mL) containing the compound from step b (1.3 g, 5.80 mmol), cyclopropanecarbonyl chloride (0.61 g, 5.80 mmol) and TEA (0.89 g, 8.70 mmol) was stirred at 0°C for 1 hour. The resulting mixture was extracted, concentrated and purified by silica gel column chromatography to give the desired compound (1 g, 58%) as a yellow solid. ESI-MS m / z: 291.85 [m+H] + .

[0499] A solution of CHCl3 (10 mL) containing the compound from step c (1 g, 3.44 mmol) and POCl3 (791 mg, 5.16 mmol) was stirred at 65°C overnight. The resulting mixture was extracted, concentrated and purified by silica gel column chromatography to give the desired compound (800 mg, 85%) as a yellow oil. ESI-MS m / z: 273.85 [m+H] + .

[0500] A solution of the compound from step d (1 g, 3.67 mmol), XantPhos (212 mg, 0.36 mmol), Ac2O (749 mg, 7.34 mmol), DIPEA (948 mg, 7.34 mmol), oxalic acid (660 mg, 7.34 mmol) and Pd(AcO)2 (82 mg, 0.367 mmol) in DMF (20 mL) was stirred at 100°C for 6 hours under nitrogen. The residue was purified by reverse phase flash chromatography to give the desired compound (587.7 mg, 61%) as a white solid. ESI-MS m / z: 238.05 [M+H] +1HNMR (400MHz, DMSO-d6) δ1.16-1.35 (m, 4H), 2.31-2.42 (m, 1H), 7.91 (t, J = 1.6Hz, 1H), 8.07-8.13 (m, 1H), 13.39 (s, 1H).

[0501] Intermediate 23

[0502]

[0503] Intermediate 23 Steps a and b

[0504]

[0505] Under nitrogen, a DMF (10 mL) solution containing 5-bromo-1,3-difluoro-2-nitrobenzene (2 g, 8.40 mmol), benzyl alcohol (4.54 g, 42.02 mmol) and CsCO (13.7 g, 42.02 mmol) was stirred overnight at 70 ° C. The aqueous layer was extracted with EA (3 x 20 mL). The combined organic layers were concentrated under reduced pressure to give a crude product, which was used directly in the next step.

[0506] A solution containing the compound from step a (2 g, 4.83 mmol) and boron trichloride (20 mL) in CH2Cl2 (40 mL) was stirred at 0°C for 2 hours. The resulting mixture was extracted with EA. The combined organic layers were washed with water. The residual product was purified by reverse phase flash chromatography to give the desired product (1.5 g, 96%) as a yellow solid. ESI-MS m / z: 322.00 [mH] - .

[0507] Intermediate 23 Steps c and d

[0508]

[0509] A solution of the compound from step b (1.5 g, 4.63 mmol), sodium 2-chloro-2,2-difluoroacetate (1.4 g, 9.26 mmol) and Cs2CO3 (3.02 g, 9.26 mmol) in DMF (15 mL) was stirred at 80° C. under N2 for 2 hours. The residue was purified by preparative TLC to give the desired product (1.2 g, 69%) as a yellow solid.

[0510] A solution of ethanol (20 mL) containing the compound from step c (1.2 g, 3.21 mmol), Fe (1.79 g, 32.07 mmol) and NH4Cl (1.72 g, 32.07 mmol) and H2O (10 mL) was stirred at 80°C under N2 atmosphere for 2 hours. The resulting mixture was filtered. The resulting mixture was extracted with EA. The combined organic layers were washed with water and the organic layer was concentrated under reduced pressure to give the desired product (1 g, 91%) as a yellow oil. ESI-MS m / z: 344.00 [m+H] + .

[0511] Intermediate 23 Steps e and f

[0512]

[0513] A solution of dioxane (20 mL) containing the compound from step d (1 g, 2.91 mmol) and cyclopropanecarbonyl chloride (607 mg, 5.81 mmol) was stirred at 100 ° C for 2 hours under N2. The resulting mixture was extracted with EA (3×2 mL). The combined organic layers were washed with water and concentrated under reduced pressure to give the desired product (900 mg, 75%) as a yellow solid. ESI-MS m / z: 412.00 [m+H] + .

[0514] A solution of DCM (30 mL) containing the compound of step e (900 mg, 2.18 mmol) and boron trichloride (11.25 mL) was stirred at 0°C for 2 hours. The resulting mixture was extracted with EA. The combined organic layers were washed with water. The residual product was purified by reverse phase flash chromatography to give the desired product (600 mg, 85%) as a yellow oil. ESI-MS m / z: 322.00 [m+H] + .

[0515] Intermediate 23 Steps g and h

[0516]

[0517] A solution of CHCl3 (10 mL) containing the compound from step f (600 mg, 1.86 mmol) and POCl3 (428 mg, 2.80 mmol) was stirred at 65°C overnight under N2. The residue was purified by silica gel column chromatography eluting with PE / EA to give the desired product (400 mg, 71%) as a yellow oil. ESI-MS m / z: 304.00 [m+H] + .

[0518] A solution of the compound from step g (400 mg, 1.32 mmol), Pd(OAc)2 (59 mg, 0.26 mmol), Ac2O (269 mg, 2.63 mmol), XantPhos (76 mg, 0.13 mmol) and DIEA (340 mg, 2.63 mm) in DMF (10 mL) was stirred at 100 °C under N2 overnight. The crude product was purified by reverse phase flash chromatography to give the desired product (200 mg, 56%) as a white solid. ESI-MS m / z: 270.00 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ1.15-1.31(m,4H),2.34(tdt,J=13.4,10.0,5.0Hz,1H),7.32-7.77(m,2H),8.01(d,J=3.0Hz,1H),13.41(s,1H).

[0519] Intermediate 24

[0520]

[0521] Intermediate 24 Step a

[0522]

[0523] A solution of 2-amino-5-bromo-3-cyclopropoxyphenol (1 g, 4.09 mmol), acetyl chloride (289 mg, 3.68 mmol) and TEA (621 mg, 6.14 mmol) in THF (10 mL) was stirred at 0°C for 2 hours. The resulting mixture was extracted, concentrated and purified by silica gel column chromatography to give the desired compound (600 mg, 51%) as a brown oil. ESI-MS m / z: 285.95 [m+H] + .

[0524] Intermediate 24 Step b

[0525]

[0526] A solution of CHCl3 (5 mL) containing the compound from step a (1.2 g, 4.19 mmol), POCl3 (1.29 g, 8.38 mmol) was stirred at 65°C for 24 hours. The resulting mixture was extracted, concentrated and purified by silica gel column chromatography to give the desired compound (1 g, 88%) as a brown solid. ESI-MS m / z: 267.90 [m+H] + .

[0527] Intermediate 24 Step c

[0528]

[0529] A solution of the compound from step b (420 mg, 1.57 mmol), Pd(OAc)2 (35 mg, 0.157 mmol), Ac2O (319 mg, 3.13 mmol), XantPhos (90 mg, 0.157 mmol), oxalic acid (282 mg, 3.13 mmol) and DIPEA (404 mg, 3.13 mmmol) in DMF (5 mL) was stirred at 100°C for 6 hours under nitrogen. The residue was purified by reverse phase flash chromatography to give the desired compound (207.3 mg, 52%) as a white solid. ESI-MS m / z: 245.90 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ0.72-0.84(m,2H),0.84-0.92(m,2H),2.62(s,3H),4 .10-4.20(m,1H),7.73(d,J=1.3Hz,1H),7.80(d,J=1.3Hz,1H),13.18(s,1H).

[0530] Intermediate 25

[0531]

[0532] Intermediate 25 Step a

[0533]

[0534] A mixture of nicotinic acid (5 g, 40.61 mmol), methoxy(methyl)amine hydrochloride (5.94 g, 60.92 mmol), HATU (15.44 g, 40.61 mmol) and DIEA (10.50 g, 81.23 mmol) in DCM (50 mL) was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was washed with water. The aqueous layer was extracted with EA. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EA (1:1) to give the desired product (3.9 g, 57%) as a light yellow liquid. ESI-MS m / z: 167.05 [m+H] + .

[0535] Intermediate Step 25b

[0536]

[0537] To a 250mL three-necked round-bottom flask was added (R)-7-bromo-5-iodo-3-methyl-2,3-dihydrofuran [2,3-c] pyridine-3-carboxamide (2 g, 5.22 mmol), the compound of step a (1.30 g, 7.83 mmol) and THF (15 mL) at room temperature. The mixture was cooled to 0°C. iPrMgCl (1.07 g, 10.44 mmol) was added dropwise under nitrogen at 0°C. The resulting mixture was stirred at room temperature for 2 hours under nitrogen. The reaction was quenched with saturated NH4Cl (aqueous solution) at room temperature. The resulting mixture was extracted with EA. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with EA to give the desired product (970 mg, 51%) as a yellow solid. ESI-MS m / z: 361.95 [m+H] + .

[0538] Intermediate 25 Step c

[0539]

[0540] Under nitrogen, a mixture of dioxane (9 mL) and H2O (1 mL) containing the compound of step b (800 mg, 2.21 mmol), 2-(5-chloro-2,4-difluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (606 mg, 2.21 mol), K2CO3 (610 mg, 4.42 mmol) and Pd(dppf)Cl2 (161 mg, 0.22 mmol) was stirred at 80 ° C for 1 hour. The resulting mixture was filtered and the filter cake was washed with EA. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EA (1:1) to give the desired product (820 mg, 86%) as a yellow solid. ESI-MS m / z: 430.05 [m+H] + .

[0541] Intermediate 25 Step d

[0542]

[0543] Under nitrogen, THF (428 mg, 3.81 mmol) containing 1M t-BuOK was added dropwise to a stirred solution of THF (50 mL) containing methyltriphenylphosphonium bromide (1.70 g, 4.77 mmol) at 0°C. The resulting mixture was stirred at 0°C for 30 minutes under nitrogen. THF containing the compound of step c (820 mg, 1.90 mmol) was added and stirred at 50°C for 2 hours. The resulting mixture was poured into water and extracted with EA. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with EA to give a crude product (430 mg) as a yellow solid. ESI-MS m / z: 428.15 [m+H] + .

[0544] Intermediate 25 Step e

[0545]

[0546] A mixture of t-BuOH (5 mL), H2O (5 ml) containing the compound from step d (400 mg, 0.93 mmol), methanesulfonamide (89 mg, 0.93 mol) and ADMIX-β (2.18 g, 2.80 mmol) was stirred at room temperature overnight. The resulting mixture was extracted with EA. The combined organic layers were concentrated under reduced pressure. The residue was purified by preparative TLC (CH2Cl2 / MeOH 10:1) to give the desired product (30 mg) as an off-white solid. ESI-MS m / z: 462.10 [m+H] + .

[0547] Intermediate 25 Step f

[0548]

[0549] A mixture of DCM (2 mL) containing the compound from step e (30 mg, 0.06 mmol), TsCl (18 mg, 0.09 mmol), TEA (19 mg, 0.19 mmol) and DMAP (8 mg, 0.06 mmol) was stirred at room temperature for 1 hour. The mixture was acidified to pH 5 with HCl (2M aqueous solution). The resulting mixture was extracted with CH2Cl2. The combined organic layers were concentrated under reduced pressure. The residue was purified by preparative TLC (EA) to give the desired product (30 mg, 74%) as a yellow solid. ESI-MS m / z: 616.15 [m+H] + .

[0550] Intermediate 25 Step g

[0551]

[0552] A solution of the compound from step f (30 mg, 0.05 mmol) and NH3(g) in MeOH (5 mL) was stirred at 40°C overnight. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (CH2Cl2 / MeOH (7M NH3) 10:1) to give the desired product (10 mg, 44%) as an off-white solid. ESI-MS m / z: 461.25 [m+H] + .

[0553] Intermediate 26

[0554]

[0555] The above compound was prepared by a similar method as Intermediate 9 to give the amino alcohol (100 mg). ESI-MS m / z: 478.50 [m+H] + .

[0556] Middle 27

[0557]

[0558] The above compound was prepared by a similar method as Intermediate 9 to give the amino alcohol. (60 mg, 53%). ESI-MS m / z: 478.20 [m+H] + .

[0559] Intermediate 28

[0560]

[0561] Intermediate 28 Step a

[0562]

[0563] A solution of (S)-(7-bromo-5-iodo-3-methyl-2,3-dihydrofuran[2,3-c]pyridin-3-yl)methanol (3 g, 7.83 mmol), 4,4,5,5-tetramethyl-2-(1-phenylethyl)-1,3,2-dioxaborolane (1.8 g, 7.83 mmol), Pd(dppf)Cl2 (573 mg, 0.78 mmol) and K2CO3 (2.1 g, 15.66 mmol) in dioxane (40 mL) and H2O (4 mL) was stirred at 80°C under nitrogen for 2 hours. The residue was purified by silica gel column chromatography eluting with PE / EA to give the desired product (2.8 g, 99%) as a brown oil. ESI-MS m / z: 346.05 [m+H] + .

[0564] Intermediate 28 Step b

[0565]

[0566] A mixture of dioxane (20 mL) and H2O (2 mL) containing the compound from step a (2 g, 5.78 mmol), 1-(5-chloro-2,4-difluorophenyl)-3,3,4,4-tetramethylborolane (1.6 g, 5.78 mol), Pd(dppf)Cl2 (422 mg, 0.58 mmol) and K2CO3 (1.6 g, 11.55 mmol) was stirred at 80°C under N2 for 2 hours. The residue was purified by silica gel column chromatography eluting with PE / EA to give the desired product (1.6 g, 66%) as a yellow solid. ESI-MS m / z: 414.20 [m+H] + .

[0567] Intermediate 28 Steps c and d

[0568]

[0569] A solution of DCM (10 mL) containing the compound of step b (1.6 g, 3.88 mmol), TBSCl (870 mg, 5.79 mmol) and imidazole (0.39 g, 5.80 mmol) was stirred at room temperature for 2 hours. The resulting mixture was extracted with DCM (3×100 mL). The combined organic layers were washed with water (3×200 mL). The organic layer was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to give the desired product (1.9 g, 93%) as a white solid. ESI-MS m / z: 528.30 [m+H] + .

[0570] A solution of t-BuOH (10 mL) and H2O (10 mL) containing the compound from step c (2 g, 3.78 mmol), ADMIX-β (8.85 g, 11.36 mmol) and methanesulfonamide (0.36 g, 3.78 mmol) was stirred at 0°C overnight. The resulting mixture was extracted with EA (3×200 mL). The combined organic layers were washed with water (3×200 mL). The organic layer was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to give the desired product (1.5 g, 70%) as a white solid. ESI-MS m / z: 562.40 [m+H] + .

[0571] Intermediate 28 Steps e and f

[0572]

[0573] A solution of DCM (20 mL) containing the compound from step d (1.5 g, 2.66 mmol), TsCl (763 mg, 4.00 mmol), TEA (810 mg, 8.00 mmol) and DMAP (232 mg, 2.66 mmol) was stirred at 0 ° C for 2 hours. The resulting mixture was extracted with EA (3×200 mL). The combined organic layers were washed with water (3×200 mL). The organic layer was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA to give the desired product (1.5 g, 78%) as a white solid. ESI-MS m / z: 716.50 [m+H] + .

[0574] A solution of the compound of step e (1.5 g, 2.66 mmol) in MeOH (5 mL) was added to a solution of NH3 (g) in MeOH (30 mL) and stirred at 40°C overnight. The residue was purified by preparative TLC (MeOH containing DCM / NH3=15 / 1) to give the desired product (1.1 g, 93%) as a white solid. ESI-MS m / z: 561.40 [m+H] + .

[0575] Intermediate 28 Steps g and h

[0576]

[0577] A solution of DCM (10 mL) containing the compound from step f (900 mg, 1.60 mmol), (Boc)2O (525 mg, 2.40 mmol) and TEA (486 mg, 4.81 mmol) was stirred at room temperature for 2 hours. The resulting mixture was extracted with DCM (3×100 mL). The combined organic layers were washed with water (3×100 mL). The organic layer was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to give the desired product (600 mg, 56%) as a white solid. ESI-MS m / z: 661.20 [m+H] + .

[0578] A solution of THF (10 mL) containing the compound from step g (900 mg, 1.36 mmol) and TBAF (355 mg, 1.36 mol) was stirred at room temperature for 1 hour. The resulting mixture was extracted with EA (3×200 mL). The combined organic layers were washed with water (3×200 mL). The organic layer was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to give the desired product (430 mg, 57%) as a white solid. ESI-MS m / z: 547.15 [m+H] + .

[0579] Intermediate 28 Steps i and j

[0580]

[0581] A solution of DCM (10 mL) containing the compound of step h (140 mg, 0.25 mmol) and DMP (271 mg, 0.64 mmol) was stirred at room temperature for 12 hours. The resulting mixture was extracted with DCM (3×20 mL). The combined organic layers were washed with water (3×20 mL). The organic layer was concentrated under reduced pressure to give the desired crude product as a white oil. ESI-MS m / z: 545.30 [m+H] + .

[0582] At room temperature, a solution of t-BuOH:H2O (2:1) containing the compound from step i (230 mg, 0.42 mmol) and 2-methyl-2-butene (354 mg, 5.04 mmol) was added dropwise to an aqueous solution of NaClO2 (380 mg, 4.20 mmol) and NaH2PO4 (504 mg, 4.20 mmol) and stirred for 2 hours. The aqueous layer was extracted with EtOAc (3x10 mL) and the resulting mixture was concentrated under reduced pressure. The crude product was used directly in the next step without further purification. ESI-MS m / z: 561.30 [m+H] + .

[0583] Intermediate 29

[0584]

[0585] Intermediate 29 Steps a and b

[0586]

[0587] A solution of the above intermediate 28 (120 mg, 0.21 mmol), deuterated methylamine (8 mg, 0.21 Mmol), HATU (81 mg, 0.21 mmmol) and DIEA (83 mg, 0.64 mmol) in DMF (1 mL) was stirred at room temperature for 2 hours. The residue was purified by preparative TLC (PE / EA 1:1) to give the desired product (80 mg, 64%) as a white solid. ESI-MS m / z: 577.15 [m+H] + .

[0588] A solution of HCl in 1,4-dioxane (2 mL) containing the compound from step a (120 mg, 2.66 mmol) was stirred at room temperature for 1 hour. The residue was purified by preparative TLC (MeOH containing DCM / NH3=15 / 1) to give the desired product (36.8 mg, 54%) as a white solid. ESI-MS m / z: 477.15 [m+H] + .

[0589] Intermediate 30

[0590]

[0591] The above compound was prepared by a method similar to that of intermediate 29 to give the amino alcohol (23 mg). ESI-MS m / z: 474.15 [m+H] + .

[0592] Intermediate 31

[0593]

[0594] The above compound was prepared in a similar manner as Intermediate 29 above to afford the amino alcohol.

[0595] Intermediate 32

[0596]

[0597] The above compound was prepared in a similar manner as Intermediate 29 above to afford the amino alcohol.

[0598] Intermediate 33

[0599]

[0600] The above compound was prepared by a similar method as Intermediate 9 to give the amino alcohol. (150 mg, 68%). ESI-MS m / z: 442.00 [m+H] + .

[0601] Intermediate 34

[0602]

[0603] The above compound was prepared by a similar method as Intermediate 9 to give the amino alcohol. (46.1 mg, 19%). ESI-MS m / z: 476.25 [m+H] + .

[0604] Intermediate 35

[0605]

[0606] The above compound was prepared by a similar method to Intermediate 9 to give amino alcohol (113.6 mg, 48%). ESI-MS m / z: 458.10 [m+H] + .

[0607] Intermediate 36

[0608]

[0609] The above compound was prepared by a similar method to Intermediate 9 to give amino alcohol (89.5 mg, 60%). ESI-MS m / z: 476.00 [m+H] + .

[0610] Intermediate 37

[0611]

[0612] The above compound was prepared in a similar manner to Intermediate 9 to afford the amino alcohol as a mixture of diastereomers. The diastereomers were separated after subsequent amide coupling and the stereochemistry was arbitrarily assigned (370 mg, 78%). ESI-MS m / z: 460.05 [m+H] + .

[0613] Intermediate 38

[0614]

[0615] Intermediate 38 Step a

[0616]

[0617] (1-(6-fluoropyridin-2-yl)vinyl)diisobutylaluminum was prepared according to Gao, F. et al., J. Am. Chem. Soc. 2010, 132, 32, 10961-10963. In a vial, (R)-7-bromo-5-iodo-3-methyl-2,3-dihydrofurano[2,3-c]pyridine-3-carboxamide (1.1 g, 2.87 mmol), xantphos (0.332 g, 0.574 mmol) and palladium (II) chloride (0.051 g, 0.287 mmol) were dissolved in DCE (3.59 ml). A THF solution containing (1-(6-fluoropyridin-2-yl)vinyl)diisobutylaluminum (5.22 ml, 5.74 mmol) was added. The vial was heated to 80 ° C and monitored by LCMS. Water was added and the aqueous layer was washed with EtOAc. The combined organic layers were dried over MgSO4 and concentrated. Purification by silica gel chromatography, 0-70% EtOAc / cHex, afforded the title compound (603 mg, 55.5%). ESI-MS m / z: 378.00 / 380.00 [m+H] + .

[0618] Intermediate 38 Step be

[0619]

[0620] The above compound was prepared in a similar manner to Intermediate 9 and the crude material was purified by preparative TLC (silica gel, DCM containing MeOH and NH3) to give the desired product (200 mg, 81%) as a white solid. ESI-MS m / z: 479.00 [m+H] + .

[0621] Intermediate 39

[0622]

[0623] The above compound was prepared in a similar manner to Intermediate 38 to give the desired product (13 mg, 62%) as a white solid. ESI-MS m / z: 495.09 [m+H] + .

[0624] Intermediate 40

[0625]

[0626] The above compound was prepared in a similar manner to Intermediate 9 and the crude material was purified by preparative TLC (silica gel, DCM containing MeOH and NH3) to give the desired product (475 mg, 71%) as a white solid. ESI-MS m / z: 466.31 [m+H]+ .

[0627] Intermediate 41

[0628]

[0629] Intermediate 41 Step a

[0630]

[0631] A solution of CH3OH (20 mL) containing 4-acetylpyridine (2 g, 16.50 mmol) and 4-toluenesulfonylhydrazide (4.61 g, 24.70 mmol) was stirred at 70°C for 2 hours. The mixture was filtered and washed to give the desired precipitated solid (2 g, 41%) as a white solid. ESI-MS m / z: 290.00 [m+H] + .

[0632] Intermediate 41 Step b

[0633]

[0634] Under nitrogen, a mixture containing the compound from step a (566 mg, 1.96 mmol), Pd(PPh3)2Cl2 (91 mg, 0.13 mmol), LiO t A solution of Bu (309 mg, 4.83 mmol) and (R)-7-bromo-5-iodo-3-methyl-2,3-dihydrofuro[2,3-c]pyridine-3-carboxamide (500 mg, 1.30 mmol) in dioxane (5 mL) was stirred at 90°C overnight. The resulting mixture was extracted, concentrated and purified by silica gel column chromatography to give the desired compound (450 mg crude product) as a yellow solid. ESI-MS m / z: 360.00 [m+H] + .

[0635] Intermediate 41 Steps c, d, e and f

[0636]

[0637] The amino alcohol was synthesized into intermediate 9 by four similar steps (cross-coupling, dihydroxylation, tosylation, amination) to give the title compound (50 mg, 74%) as a white solid. ESI-MS m / z: 460.95 [m+H] + .

[0638] After subsequent amide coupling, the diastereomers were separated using preparative HPLC.

[0639] Intermediate 42

[0640]

[0641] Intermediate 42 Step a

[0642]

[0643] A mixture of methanol (18 mL) containing p-(trifluoromethyl)acetophenone (1 g, 5.32 mmol) and TsNHNH2 (1.48 g, 8.00 mmol) was stirred at 70 ° C for 5 hours. The reaction was monitored by TLC. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (1:1) to give the desired compound (1.75 g, 92%) as a white solid. ESI-MS m / z: 357.05 [m+H] + .

[0644] Intermediate 42 Step b

[0645]

[0646] At room temperature, the compound from step a (1.29 g, 3.62 mmol), TMBQ (1.63 g, 10.86 mmol), biboronic acid pinacol ester (1.38 g, 5.43 mmol), Pd (OAc) 2 (41 mg, 0.18 mmol), PPh 3 (95 mg, 0.36 mmol) and NaH (261 mg, 10.86 mmol) were added to a 40 mL vial. After degassing and filling with N 2 , toluene (40 mL) was added to the vial. The reaction mixture was reacted overnight at 90 ° C. The reaction was monitored by LCMS. The resulting mixture was filtered and the filter cake was washed with ethyl acetate. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (1: 1) to obtain the desired compound (2.4 g) as a brown solid. 1 H NMR (400MHz, CDCl3) δ1.33 (s, 12H), 6.13 (s, 1H), 6.17 (d, J = 2.7Hz, 1H), 7.57 (s, 4H).

[0647] Intermediate 42 Step c

[0648]

[0649] Under nitrogen, a solution of dioxane (8 mL) and H2O (2 mL) containing the compound of step b (778 mg, 2.61 mmol) was treated with (R)-7-bromo-5-iodo-3-methyl-2,3-dihydrofuran[2,3-c]pyridine-3-carboxamide (500 mg, 1.31 mmol), K2CO3 (361 mg, 2.61 mmol) and Pd(dppf)Cl2CH2Cl2 (106 mg, 0.13 mmol) at 80°C for 1 hour. The reaction was monitored by LCMS. The resulting mixture was extracted with EtOAc. The combined organic layers were concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography to give the desired compound (443 mg, 79%) as a brown oil. ESI-MS m / z:427.00[m+H] + .

[0650] Intermediate 42 Steps d, e, f and g

[0651]

[0652] The amino alcohol was synthesized from intermediate 9 by four similar steps (cross coupling, racemic dihydroxylation, tosylation, amination) to give the title compound (208 mg, 90%) as a white solid. ESI-MS m / z: 528.10 [m+H] + .

[0653] After subsequent amide coupling, the diastereomers were separated using preparative TLC.

[0654] Intermediate 43

[0655]

[0656] Selective dihydroxylation using ADMIX-β and the synthesis of the amino alcohol in a similar sequence to intermediate 42 gave the title compound (181.4 mg, 66%) as a white solid. ESI-MS m / z: 528.10 [m+H] + .

[0657] Intermediate 44

[0658]

[0659] Selective dihydroxylation using ADMIX-β and synthesis of the amino alcohol in a similar sequence to intermediate 42 afforded the title compound (10 mg, 23%) as a white solid. ESI-MS m / z: 464.30 [m+H] + .

[0660] Intermediate 45

[0661]

[0662] Intermediate 45 Step a

[0663]

[0664] (R)-7-Bromo-5-(1-cyclopropylvinyl)-3-methyl-2,3-dihydrofurano[2,3-c]pyridine-3-carboxamide (1.42 g, 4.39 mmol, 1.00 equiv) dissolved in 1,4-dioxane (22 mL) and sodium iodide (2.31 g, 15.4 mmol, 3.50 equiv) was added. The suspension was sparged with nitrogen and copper(I) iodide (167 mg, 0.879 mmol, 0.2 equiv) and N,N'-dimethylethylenediamine (0.208 mL, 1.93 mmol, 0.4 equiv) were added. Sparging was continued for another 5 minutes and then a reflux condenser was installed on the flask and heated to 100°C with stirring. After stirring at this temperature for 2 hours, the reaction mixture was cooled to room temperature and diluted with saturated aqueous sodium bicarbonate, water and EtOAc. The layers were separated and the aqueous layer was extracted with EtOAc (3×50 mL). The combined organic extracts were dried over sodium sulfate, filtered, and the filtrate was concentrated to a crude oil. The crude material was purified by automated flash chromatography on silica gel (containing 0-100% EtOAc in cyclohexane) to give the pure iodide product (1.03 g, 63%) as a yellow foam. ESI-MS m / z: 371.178 [m+H] + .

[0665] Intermediate 45 Step b

[0666]

[0667] 1-Bromo-2,3-dichloro-4-fluorobenzene (4.74 g, 19.4 mmol) was dissolved in THF (97 ml) and the solution was cooled to -40 ° C in a dry ice / acetone bath. Isopropylmagnesium chloride (2M in THF, 10.7 ml, 21.4 mmol, 1.1 equiv) was added and the resulting mixture was stirred at this temperature for one hour. After that, i-PrOB(pin) (4.76 ml, 23.3 mmol, 1.2 equiv) was added. The reaction mixture was then heated to room temperature and stirred. During this period, some solids began to precipitate. After 1 hour, the reaction was quenched with saturated aqueous sodium bicarbonate. The layers were separated and the aqueous layer was extracted with EtOAc (3×50 mL). The combined organic extracts were washed with brine (1×50 mL), dried over sodium sulfate, filtered and concentrated to give the crude borate (5.65 g, 89%) as a crystalline white solid. The crude product contained approximately 8 wt% pinacol and was used without further purification (no m / z for the boronate ester was detected).

[0668] Intermediate 45 Step c

[0669]

[0670] The compound of step a (1.04 g, 2.81 mmol, 1.0 equiv) was dissolved in 1,4-dioxane (11.2 mL) and K2CO3 (1.17 g, 8.43 mmol, 3.0 equiv), Pd(dppf)Cl2 (123 mg, 0.169 mmol, 0.06 equiv) and the boronate ester of step b (1.10 g, 3.79 mmol, 1.35 equiv) were added. Water (2.8 mL) was added and the resulting two-phase suspension was bubbled with nitrogen for 5 minutes. The vial was sealed and heated to 90 °C with stirring for one hour and then cooled to room temperature. The reaction mixture was diluted with water and EtOAc. The layers were separated and the aqueous layer was extracted with EtOAc (3×50 mL). The combined organic extracts were dried over sodium sulfate, filtered, and the filtrate was concentrated to a crude oil. The crude material was purified by automated flash chromatography on silica gel (0-100% EtOAc in cyclohexane) to give the pure product 5 (940 mg, 82%) as a yellow foam. ESI-MS m / z: 407.156 [m+H] + .

[0671] Intermediate 45 Steps d, e and f

[0672]

[0673] The above compound was prepared by a similar procedure as Intermediate 9 to give the desired product (700 mg, 99%) as a white solid. ESI-MS m / z: 440.08 [m+H] + .

[0674] Intermediate 46

[0675]

[0676] The above compound was prepared in a similar manner to Intermediate 45 to give the desired product (138 mg, 90%) as a white solid. ESI-MS m / z: 504.14 [m+H] + .

[0677] Intermediate 47

[0678]

[0679] The above compound was prepared in a similar manner to Intermediate 45 to give the desired product (417 mg, 99%) as a white solid. ESI-MS m / z: 476.17 [m+H] + .

[0680] Intermediate 48

[0681]

[0682] The above compound was prepared in a similar manner to Intermediate 45 to give the desired product (222 mg, 93%) as a white solid. ESI-MS m / z: 458.22 [m+H] + .

[0683] Intermediate 49

[0684]

[0685] The above compound was prepared in a similar manner to Intermediate 45 to give the desired product (158 mg, 100%) as a white solid. ESI-MS m / z: 522.18 [m+H] + .

[0686] Intermediate 50

[0687]

[0688] The above compound was prepared in a similar manner to Intermediate 45 to give the desired product (291 mg, 76%) as a white solid. ESI-MS m / z: 494.24 [m+H] + .

[0689] Intermediate 51

[0690]

[0691] The above compound was prepared in a similar manner to Intermediate 45 to give the desired product (158 mg, 100%) as a white solid. ESI-MS m / z: 522.18 [m+H] + .

[0692] Intermediate 52

[0693]

[0694] The above compound was prepared in a similar manner to Intermediate 45 to give the desired product (800 mg, 96%) as a white solid. ESI-MS m / z: 476.17 [m+H] + .

[0695] Intermediate 53

[0696]

[0697] The above compound was prepared by a similar procedure as Intermediate 9 to give the desired product (421 mg, 90%) as a white solid. ESI-MS m / z: 440.08 [m+H] + .

[0698] Intermediate 54

[0699]

[0700] The above compound was prepared by a similar procedure as Intermediate 9 to give the desired product (608 mg, 97%) as a white solid. ESI-MS m / z: 440.08 [m+H] + .

[0701] Intermediate 55

[0702]

[0703] The above compound was prepared by a similar procedure as Intermediate 9 to give the desired product (560 mg, 93%) as a white solid. ESI-MS m / z: 495.45 [m+H] + .

[0704] Intermediate 56

[0705]

[0706] The above compound was prepared in a manner similar to Intermediate 9 to give the desired amino alcohol (689 mg, 94%) as an off-white solid. ESI-MS m / z: 459.44 [m+H] + .

[0707] Intermediate 58

[0708]

[0709] The above compound was prepared in a similar manner to Intermediate 9 to give the desired amino alcohol (293 mg, 100%) as an off-white solid. ESI-MS m / z: 441.34 [m+H] + .

[0710] Table 3 below contains examples prepared in a manner similar to Example 1. Most compounds were purified by preparative HPLC (ACN / H2O, 20-90%, 25 minutes) and some compounds were purified by automated column chromatography (silica gel). Aryl boronate starting materials and aryl amide coupling agents were prepared according to Intermediates 1-58, or by similar methods with slight modifications, also according to the method in U.S. Pat. No. 11,572,367.

[0711] Table 3

[0712]

[0713]

[0714]

[0715]

[0716]

[0717]

[0718]

[0719]

[0720]

[0721]

[0722]

[0723]

[0724]

[0725]

[0726]

[0727]

[0728]

[0729]

[0730] Table 4 below contains examples prepared using a method similar to Example 47. Most compounds were purified by preparative HPLC (ACN / H2O, 20-90%, 25 minutes) and some compounds were purified by automated column chromatography (silica gel). Some examples could not be separated by preparative HPLC and are reported as a mixture of diastereomers.

[0731] Table 4

[0732]

[0733]

[0734]

[0735] Intermediate 59

[0736]

[0737] Intermediate 59 Step a

[0738]

[0739] A solution of CAN-7-bromo-5-iodo-3-methyl-2,3-dihydrofuran[2,3-c]pyridine-3-carboxamide (2 g, 5.22 mmol), Pd(dppf)Cl2 CH2Cl2 (0.43 g, 0.52 mmol) and K2CO3 (1.44 g, 10.44 mmol) and tert-butyl N-[2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)prop-2-en-1-yl]carbamate (1.48 g, 5.22 mmol) in dioxane (16 mL), H2O (4 mL) was stirred at 80°C for 2 hours under nitrogen. The resulting mixture was extracted, concentrated and purified by silica gel column chromatography to give the desired compound (1.9 g, 88%) as a yellow oil. ESI-MS m / z: 413.90 [m+H] + .

[0740] Intermediate 59 Step b

[0741]

[0742] Under nitrogen, a solution of dioxane (18 mL) and H2O (2 mL) containing the compound from step a (1.9 g, 4.61 mmol), Pd(dppf)Cl2CH2Cl2 (375 mg, 0.46 mmol) and K2CO3 (1.27 g, 9.22 mmol) and 2-(5-chloro-2,4-difluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.90 g, 6.92 mmol) was stirred at 80°C for 2 hours. The resulting mixture was extracted, concentrated and purified by silica gel column chromatography to give the desired compound (1.8 g, 81%) as a yellow solid. ESI-MS m / z: 480.10 [m+H] + .

[0743] Intermediate 59 Step c

[0744]

[0745] A solution of acetone (20 ml) containing the compound from step b (2 g, 4.16 mmol), K2OsO4·2H2O (0.06 g, 0.17 mmol), NMO (1.37 g, 11.67 mmol) and H2O (20 mL) was stirred at room temperature overnight. The resulting mixture was extracted, concentrated and dissolved in H2O (20 mL) and acetone (20 ml). NaIO4 (4.01 g, 18.75 mmol) was added. The resulting mixture was stirred at room temperature for another 4 hours. The resulting mixture was extracted, concentrated and purified by silica gel column chromatography to give the desired compound (1.7 g, 84%) as a yellow solid. ESI-MS m / z: 482.00 [m+H] + .

[0746] Embodiment 463

[0747]

[0748] Example 463 Step a

[0749]

[0750] Under nitrogen, a solution of THF (3 mL) containing 2-bromopyridine (373 mg, 2.36 mmol) and n-BuLi (151 mg, 2.36 mmol) was stirred at -78 ° C for 15 minutes. Intermediate 59 (190 mg, 0.39 mmol) was added dropwise at -78 ° C. The resulting mixture was stirred for another 30 minutes at -78 ° C. The residue was quenched, extracted, concentrated, and purified by reverse phase flash chromatography to give the desired compound (100 mg, 45%) as a yellow crude solid. ESI-MS m / z: 561.05 [m+H] + .

[0751] Example 463 Step b

[0752]

[0753] A solution containing the compound from step a (100 mg, 0.18 mmol), 1,4-dioxane (2 mL) containing HCl and DCM (2 ml) was stirred at room temperature for 1 hour. The mixture was basified, extracted, concentrated and purified by reverse phase flash chromatography to give the desired compound (50 mg, 60%) as a white solid. ESI-MS m / z: 460.95 [m+H] + .

[0754] Example 463 Step c

[0755]

[0756] A solution of DMF (1 mL) containing the compound from step b (50 mg, 0.11 mmol), HATU (61 mg, 0.16 mmol) and DIPEA (42 mg, 0.32 mmol) and 2-cyclopropyl-7-methoxy-2H-indazole-5-carboxylic acid (25 mg, 0.11 mmol) was stirred at room temperature for 1 hour. The resulting mixture was extracted, concentrated and purified by preparative HPLC to give the desired compound (3.5 mg, 17%) as a white solid. ESI-MS m / z: 675.20 [m+H] + .

[0757] Intermediate 60

[0758]

[0759] The following intermediate was prepared in a similar order to Example 463 step b to give the title compound (110 mg, 64%). ESI-MS m / z: 463.95 [m+H] + .

[0760] Intermediate 61

[0761]

[0762] The following intermediate was prepared in a similar order to Example 463 step b to give the title compound (63 mg) ESI-MS m / z: 486.15 [m+H] + .

[0763] Table 7 below contains examples prepared in analogy to Example 463. Most compounds were purified by preparative HPLC (ACN / H2O, 20-90%, 25 min) and some were purified by automated column chromatography (silica gel).

[0764] Table 7

[0765]

[0766]

[0767] Intermediate 62

[0768]

[0769] Intermediate 62 Step a

[0770]

[0771] To a stirred solution of CAN-7-bromo-5-iodo-3-methyl-2,3-dihydrofuran[2,3-c]pyridine-3-carboxamide (4 g, 10.44 mmol) and tributyl(1-ethoxyvinyl)stannane (3.96 g, 10.97 mmol) in toluene was added dropwise Pd(dppf)Cl2 CH2Cl2 (851 mg, 1.04 mmol) at 110°C under nitrogen for 2 hours. The resulting mixture was extracted with EA. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (1:1) to give the desired compound (2.1 g, 67%) as a brown solid. ESI-MS m / z: 327.00 [m+H] + .

[0772] Intermediate 62 Step b

[0773]

[0774] To a stirred solution of dioxane (40 mL) containing the compound from step a (4.7 g, 14.37 mmol) and 2-(5-chloro-2,4-difluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3.94 g, 14.37 mol) was added H2O (10 mL), Pd(dppf)Cl2CH2Cl2 (1.17 g, 1.44 mmol) and K2CO3 (3.97 g, 28.73 mmol). The resulting mixture was stirred at 80 °C for 2 hours under nitrogen. The reaction was monitored by LCMS. The resulting mixture was extracted with EA. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE / EA (5:1) to give the desired compound (4 g, 71%) as a yellow oil. ESI-MS m / z: 395.05 [m+H] + .

[0775] Intermediate 62 Step c

[0776]

[0777] Under nitrogen atmosphere, a MeOH (20 mL) solution containing the compound of step b (6 g, 16.36 mmol) and HCl (33 mL) was stirred at 50 ° C for 2 hours. The resulting mixture was concentrated under reduced pressure. The mixture was acidified to pH 7 with saturated NaHCO3 (aqueous solution). The resulting mixture was extracted with CH2Cl2 (3×30 mL). The combined organic layer was washed with brine (1×10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (5:1) to obtain the desired compound (4 g, 64%) as a yellow oil. ESI-MS m / z:381.90[m+H] + .

[0778] Intermediate 62 Step d

[0779]

[0780] The compound of step c (4 g, 10.48 mmol) was dissolved in DCM (50 mL) at 0 ° C. 2,6-di-tert-butyl-4-methylpyridine (2.58 g, 12.57 mmol) and Tf2O (4.43 g, 15.72 mmol) were added. The reaction mixture was heated to room temperature and stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The resulting mixture was extracted with EtOAc. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (5: 1) to give the title compound (4 g, 74%) as a yellow oil. ESI-MS m / z: 514.00 [m+H] + .

[0781] Intermediate 63

[0782]

[0783] The following intermediate was prepared in a similar manner to Intermediate 62 to give the desired compound (1.7 g, 65%). ESI-MS m / z: 512.00 [m+H] + .

[0784] Intermediate 64

[0785]

[0786] Intermediate 64 Step a

[0787]

[0788] To a stirred solution of 1,4-dioxane / H2O (9:1) containing intermediate 62 (200 mg, 0.39 mmol) and 2-cyclopropylpyrimidin-5-ylboronic acid (64 mg, 0.39 mmol) were added K2CO3 (108 mg, 0.78 mmol) and Pd(dppf)Cl2CH2Cl2 (32 mg, 0.04 mmol), and the resulting mixture was stirred at 85°C under nitrogen for 2 hours. The residue was purified by reverse phase flash chromatography to give the desired compound (130 mg, 69%) as a yellow oil. ESI-MS m / z: 484.30 [m+H] + .

[0789] Intermediate 64 Step b

[0790]

[0791] ADMIX-β (1.16 g, 1.49 mmol) was added to a stirred solution of t-BuOH / H2O containing the compound of step a (240 mg, 0.50 mmol) and methanesulfonamide (47 mg, 0.50 mmol), and the resulting mixture was stirred at room temperature overnight. The mixture was extracted with EtOAc (3x10 mL). The residue was purified by reverse phase flash chromatography to give the desired compound (115 mg, 45%) as a yellow oil. ESI-MS m / z: 518.15 [m+H] + .

[0792] Intermediate 64 Steps c and d

[0793]

[0794] Under nitrogen, TEA (67 mg, 0.67 mmol) and DMAP (27 mg, 0.22 mmol) were added to a stirred solution of DCM containing the compound of step b (100 mg, 0.20 mmol) and TsCl (64 mg, 0.33 mmol) at room temperature and stirred for 2 hours. The resulting mixture was acidified to pH 3 with HCl (1 M aqueous solution), extracted with DCM, and the combined organic layers were concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA 1:1) to give the desired compound (80 mg, 62%) as a yellow solid. ESI-MS m / z: 672.25 [m+H] + .

[0795] A solution of the compound from step c (80 mg, 0.12 mmol) and NH3(g) in MeOH (10 mL) was stirred at 40°C overnight. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (MeOH containing CH2Cl2 / NH3 at 12:1) to give the methyl title compound (60 mg, 130%) as a yellow solid. ESI-MS m / z: 502.25 [m+H] + .

[0796] Intermediate 65

[0797]

[0798] The following intermediate was prepared in a similar order to Intermediate 64 to give the title compound (110 mg, 64%). ESI-MS m / z: 444.00 [m+H] + .

[0799] Intermediate 66

[0800]

[0801] The following intermediate was prepared in a similar sequence to Intermediate 64 to give the title compound.

[0802] Intermediate 67

[0803]

[0804] The following intermediate was prepared in a similar sequence to Intermediate 64 to give the title compound.

[0805] Intermediate 68

[0806]

[0807] The following intermediate was prepared in a similar sequence to Intermediate 64 to give the title compound.

[0808] Intermediate 69

[0809]

[0810] The following intermediate was prepared in a similar sequence to Intermediate 64 to give the title compound (50.2 mg, 41%) as a white solid. ESI-MS m / z: 538.15 [m+H] + .

[0811] Intermediate 70

[0812]

[0813] The following intermediate was prepared in a similar sequence to Intermediate 64 to give the title compound.

[0814] Intermediate 71

[0815]

[0816] The following intermediate was prepared in a similar order to Intermediate 64 to give the title compound (70 mg, 75%) as a white solid. ESI-MS m / z: 576.25 [m+H] + After subsequent amide coupling, the final compound was isolated after deprotection of Boc with dioxane containing HCl. See the table below.

[0817] Intermediate 72

[0818]

[0819] The following intermediate was prepared in a similar sequence to Intermediate 64 to give the title compound (80 mg, 65%) as a white solid. ESI-MS m / z: 590.00 [m+H] + After subsequent amide coupling, the final compound was isolated after deprotection of Boc using dioxane containing HCl. See the table below.

[0820] Intermediate 73

[0821]

[0822] The following intermediate was prepared in a similar sequence to Intermediate 64 to give the title compound (27 mg, 42%) as a white solid. ESI-MS m / z: 474.95 [m+H] + .

[0823] Intermediate 74

[0824]

[0825] The following intermediate was prepared in a similar order to Intermediate 64 to give the title compound (70 mg) as a white solid. ESI-MS m / z: 639.00 [m+H] + After subsequent amide coupling, the final compound was isolated after deprotection of Boc using dioxane containing HCl. See the table below.

[0826] Intermediate 75

[0827]

[0828] The following intermediate was prepared in a similar sequence to Intermediate 64 to give the title compound (70 mg, 71%) as a yellow solid. ESI-MS m / z: 484.95 [m+H] + .

[0829] Intermediate 76

[0830]

[0831] The following intermediate was prepared in a similar sequence to Intermediate 64 to give the title compound (30 mg, 81%) as a yellow solid. ESI-MS m / z: 495.00 [m+H] + .

[0832] Intermediate 77

[0833]

[0834] The following intermediate was prepared in a similar order to Intermediate 64 to give the title compound (22 mg) as a yellow solid. ESI-MS m / z: 451.05 [m+H] + .

[0835] Intermediate 78

[0836]

[0837] The following intermediate was prepared in a similar sequence to Intermediate 64 to give the title compound (59.4 mg, 69%) as a yellow solid. ESI-MS m / z: 509.10 [m+H] + .

[0838] Intermediate 79

[0839]

[0840] Intermediate 79 Step a

[0841]

[0842] A DMF (10 mL) solution containing compound A3-1036 (280 mg, 0.55 mmol), Pd(dppf)Cl2 (40 mg, 0.06 mmol), CuCl (54 mg, 0.55 mmol) and Cs2CO3 (355 mg, 1.09 mmol) was stirred at 90°C under N2 for 2 hours. The residue was purified by silica gel column chromatography eluting with PE / EA to give the desired product (70 mg, 29%) as a yellow oil. ESI-MS m / z: 444.00 [m+H] + .

[0843] Intermediate 79 Steps b, c, d and e

[0844]

[0845] The following intermediate was prepared in a similar sequence to Intermediate 64 to give the title compound (15 mg, 68%) as a yellow solid. ESI-MS m / z: 462.00 [m+H] + .

[0846] Table 8 below contains examples prepared in a manner similar to Example 1. Most compounds were purified by preparative HPLC (ACN / H2O , 20-90%, 25 minutes) and some compounds were purified by automated column chromatography (silica gel). Amine starting materials were prepared according to Intermediates 62-80, and aromatic amide coupling agents were prepared according to Intermediates 1-58, or by similar procedures with slight modifications according to the procedures in U.S. Pat. No. 11572367.

[0847] Table 8

[0848]

[0849]

[0850]

[0851] Embodiment 504

[0852]

[0853] Example 504 Step a

[0854]

[0855] To a stirred solution of 1,4-dioxane / H2O containing intermediate 62 (300 mg, 0.58 mmol) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyridin-2-one (129 mg, 0.58 mol) was added K2CO3 (161 mg, 1.17 mmol) and Pd(dppf)Cl2 CH2Cl2 (47 mg, 0.06 mmol) and the reaction was stirred at 85°C under nitrogen for 2 hours. The residue was purified by reverse phase flash chromatography to give the desired product (105 mg, 39%) as a yellow solid. ESI-MS m / z: 459.00 [m+H] + .

[0856] Example 504 Step b

[0857]

[0858] A mixture of tBuOH (3 mL), H2O (3 mL) containing the methyl compound from step a (100 mg, 0.22 mmol), methanesulfonamide (20 mg, 0.22 mmol) and ADMIX-β (509 mg, 0.65 mmol) was stirred at room temperature overnight. The reaction was monitored by LCMS. The resulting mixture was extracted with EA. The combined organic layers were concentrated under reduced pressure. The residue was purified by preparative TLC (EA) to give the desired product (70 mg, 65%) as a yellow-brown oil. ESI-MS m / z: 493.15 [m+H] + .

[0859] Example 504 Step c

[0860]

[0861] A mixture of DCM (3 mL) containing the compound from step b (70 mg, 0.14 mmol), TsCl (40 mg, 0.21 mmol), DMAP (17 mg, 0.14 mol) and TEA (43 mg, 0.43 mmol) was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The mixture was acidified to pH 4 with HCl (1M aqueous solution). The resulting mixture was extracted with CH2Cl2. The combined organic layers were concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA 1:1) to give the desired product (60 mg, 52.73%) as a yellow solid. ESI-MS m / z: 800.95 [m+H] + .

[0862] Example 504 Step d

[0863]

[0864] A mixture of the compound from step c (60 mg, 0.07 mmol) and NH3(g) in MeOH (3 mL) was stirred at 40°C overnight. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (containing CH2Cl2 / 7M NH3 in MeOH) to give the desired product (20 mg, 42.3%) as a white solid. ESI-MS m / z: 631.15 [m+H] + .

[0865] Example 504 Step e

[0866]

[0867] A mixture of DMF (1 mL) containing the compound from step d (25 mg, 0.04 mmol), 2-cyclopropyl-7-methoxy-2H-indazole-5-carboxylic acid (9 mg, 0.04 mmol), HATU (15 mg, 0.04 mmol) and DIEA (10 mg, 0.08 mmol) was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The residue was purified by reverse phase flash chromatography to give the desired product (20 mg, 59%) as a yellow solid. ESI-MS m / z: 845.05 [m+H] + .

[0868] Example 504 Step f

[0869]

[0870] To a stirred mixture containing the compound of step e (20 mg, 0.02 mmol) and MeOH (2 mL) was added a solution of LiOH (5 mg, 0.24 mmol) in H2O (0.5 mL) at room temperature. The reaction was monitored by LCMS. The resulting mixture was concentrated under vacuum. The residue was purified by preparative TLC (MeOH containing CH2Cl2 / 7M NH3 at 10:1) to give the desired product (6.7 mg, 38%) as a white solid. ESI-MS m / z: 691.20 [m+H] + .

[0871] Embodiment 505

[0872]

[0873] The following examples were prepared in a similar order to Example 504. The residue was purified by reverse phase flash chromatography to give the title compound (6.4 mg, 31%) as a white solid. ESI-MS m / z: 691.20 [m+H] + .

[0874] Test

[0875] RSV-A test method

[0876] Hep-2 cells (originally derived from tumors growing in irradiated cortisolized weanling rats injected with epidermoid carcinoma tissue from the larynx of a 56-year-old male, but later found to be indistinguishable from HeLa cells by PCRDNA analysis) were used to culture genotype A "long" strain RSV. Flasks were inoculated with RSV and virus stocks were harvested once the cytopathic effect (CPE) was greater than 90%. Virus stocks were snap-frozen in 25% sucrose medium using liquid nitrogen to improve virus stability. Virus stock titers were measured by tissue culture infectious dose 50% (TCID50 ), 3-fold virus dilutions were used at 8,000 cells per well in 96-well plates and quantified for 4 days. Virus stock titers were also quantified by plaque forming unit assay as described elsewhere.

[0877] After extensive parameter testing, the final assay was performed as follows: Hep-2 cells were seeded at 8,000 cells per well in a 50 μL volume into the inner 60 wells of a 96-well plate using growth medium (DMEM without phenol red, 1% L-Glut, 1% Penn / Strep, 1% non-essential amino acids, 10% heat-inactivated FBS). Serial 2-fold dilutions of control and test compounds were added to the wells in duplicate for a total volume of 25 μL. The virus stock was then added to the wells at a multiplicity of infection (MOI) of 0.1 in a volume of 25 L, bringing the total volume of each well to 100 L. MOI was expressed in PFU / mL or TCID 50 (if PFU / mL is not available). Each 96-well plate has a 6-well control column containing cells and virus but no compound (negative control, maximum CPE), a column containing cells but no compound or virus (positive control, minimum CPE), and a column with no cells, virus, or compound (background plate / reagent control). Control wells with cells but no virus are given an additional 25 μL of growth media containing an equal amount of sucrose as the wells receiving the virus stock to keep media and volume conditions consistent. The outer wells of the plate are filled with 125 μL of moat media (DMEM, 1% Penn / Strep) to act as a thermal and evaporative moat around the test wells. After a 5-day incubation period, the plates are read using ATPlite (50uL added per well), which quantifies the amount of ATP present in each well (a measure of cell health). The assay plates are read using an Envision photometer. These data are used to calculate the EC for each compound. 50 (Table 10). 50 The ranges are as follows: A < 0.2 μM; B > 0.2M.

[0878] Table 10 Summary of RSV-A activity

[0879]

[0880]

[0881]

[0882]

[0883]

[0884]

[0885] HMPV antiviral detection method

[0886] Method 1:

[0887] In vitro HMPV antiviral activity was assessed using the clinical isolate A2 strain TN 94-49 (obtained from Dr. John Williams, Vanderbilt University, Tennessee) and LLC-MK2 cells (ATCC #CCL-7), an immortalized kidney epithelial cell line from rhesus monkeys.

[0888] The compounds were resuspended in 10 mM dimethyl sulfoxide (DMSO), serially diluted and added to a 384-well source plate. Subsequently, the compounds were diluted and transferred to a 384-well assay plate using an Echo-650 automated liquid handling system (Beckman Coulter, Indiana). The test compounds were evaluated twice at a maximum concentration of 2 μM, followed by 2.5-fold serial dilutions to give a total of 10 concentration points. DMSO control wells were also included on the assay plate, either infected or uninfected, as positive and negative controls.

[0889] A2 TN 94-49 virus infection was performed in LLC-MK2 cell suspension. The cells were washed twice with PBS and removed from the cell culture flask with 0.25% trypsin-EDTA (Thermo Fisher Scientific, Massachusetts). Trypsin-EDTA was inactivated by resuspending in 2% fetal bovine serum (FBS) and OptiMEM (Thermo Fisher Scientific, Massachusetts) containing 1% penicillin-streptomycin. The cells were precipitated by centrifugation at 800 rpm for 5 minutes, the supernatant was removed, the cells were resuspended and washed in PBS with 100 μg / mL CaCl2. This step was performed twice. The cells were then resuspended in serum-free (SF)-OptiMEM containing 4 μg / mL TPCK trypsin (Sigma-Aldrich, Missouri), 1% penicillin-streptomycin (ThermoFisher Scientific, Massachusetts) and 100 μg / mL CaCl2. Cells were counted and seeded at a density of 5000 cells / well, 12.5 μL / well.

[0890] Virus infection was performed at a multiplicity of infection (MOI) of 0.005, with 12.5 L added to each well. Virus infection was performed in infection medium containing SF-OptiMEM, 100 μg / mL CaCl2, and 1% penicillin-streptomycin. HMPV virus stock was suspended in infection medium + 5% glycerol, so an equal volume of infection medium + 5% glycerol was added to uninfected wells to balance the final glycerol percentage of all wells of the assay plate. The final concentration of TPCK trypsin was 2 μg / mL. The assay plate was incubated at 37°C, 5% CO2 for 7 days.

[0891] After 7 days of incubation, 12.5 μL of ATP-Lite (PerkinElmer, MA) was added to each well and raw luminescence values ​​were determined using Envision 2104 (Perkin-Elmer, MA). The average of the raw luminescence values ​​for the cell and virus-only positive control wells was subtracted from all tested conditions, and the percentage of cell health was determined by dividing these values ​​by the average of the cell-only negative control wells. EC was then calculated by nonlinear regression using a four-parameter logistic equation. 50 The curve fitting model used was the XLFit dose-response one-site model 200:

[0892] y = (A + (B / (1 + ((x / C) ^ D)))), where A is the minimum y value, B is the maximum y value, and C is the logEC 50 value, and D is the slope factor.

[0893] Method 2:

[0894] Evaluation of in vitro HMPV antiviral activity using clinical isolate A2 strain TN 94-49

[0895] In vitro HMPV antiviral activity was assessed using the clinical isolate A2 strain TN 94-49 (obtained from Dr. John Williams, Vanderbilt University, Tennessee) and LLC-MK2 cells (Millipore Sigma CB_85062804), an immortalized kidney epithelial cell line from macaques.

[0896] The compounds were resuspended in 10 mM dimethyl sulfoxide (DMSO), serially diluted and added to a 384-well source plate. Subsequently, the compounds were diluted and transferred to a 384-well assay plate using an Echo-650 automated liquid handling system (Beckman Coulter, Indiana). The test compounds were evaluated twice with a maximum concentration of 200 nM followed by 2.5-fold serial dilutions for a total of 10 concentration points. DMSO control wells were also included on the assay plate, either infected or uninfected, as positive and negative controls.

[0897] A2 TN 94-49 virus infection was performed in LLC-MK2 cell suspension. The cells were washed twice with PBS and removed from the cell culture flask with 0.25% trypsin-EDTA (Thermo Fisher Scientific, Massachusetts). Trypsin-EDTA was inactivated by resuspending in 2% fetal bovine serum (FBS) and OptiMEM (ThermoFisher Scientific, Massachusetts) containing 1% antibiotic antimycotic. The cells were precipitated by centrifugation at 800 rpm for 5 minutes, the supernatant was removed, the cells were resuspended and washed in PBS with 100 μg / mL CaCl2. This step was performed twice. The cells were then resuspended in serum-free (SF)-OptiMEM containing 4 μg / mL TPCK trypsin (Sigma-Aldrich, Missouri), 1% antibiotic antimycotic (ThermoFisher Scientific, Massachusetts), 100 μg / mL CaCl2 and 4 μM P-glycoprotein inhibitor CP-100-356. Cells were counted and seeded at a density of 5000 cells / well, 12.5 μL / well.

[0898] Virus infection was performed at a multiplicity of infection (MOI) of 0.16, with 12.5 μL added to each well. Virus infection was performed in infection medium containing SF-OptiMEM, 100 μg / mL CaCl2, and 1% antibiotic antimycotic. HMPV virus stock was suspended in infection medium + 5% glycerol, so an equal volume of infection medium + 5% glycerol was added to the uninfected wells to balance the final glycerol percentage of all wells of the assay plate. The final concentration of TPCK trypsin was 2 μg / mL. The final concentration of CP-100-356 was also 2 μM. The assay plate was incubated at 37°C, 5% CO2 for 7 days.

[0899] After 7 days of incubation, 12.5 μL of ATP-Lite (PerkinElmer, MA) was added to each well and raw luminescence values ​​were determined using Envision 2104 (Perkin-Elmer, MA). The average of the raw luminescence values ​​for the cell and virus-only positive control wells was subtracted from all tested conditions, and the percentage of cell health was determined by dividing these values ​​by the average of the cell-only negative control wells. EC was then calculated by nonlinear regression using a four-parameter logistic equation. 50 The curve fitting model used was the XLFit dose-response one-site model 200:

[0900] y = (A + (B / (1 + ((x / C) ^ D)))), where A is the minimum y value, B is the maximum y value, and C is the logEC50 value, and D is the slope factor.

[0901] These data were used to calculate the EC for each compound. 50 (Table 11).

[0902] Method 1EC 50 The ranges are as follows: A < 0.5 μM; B > 0.5 μM. EC of Method 2 50 The ranges are as follows: A < 0.2 μM; B > 0.2 μM.

[0903] Table 11 Summary of the activities of hMPVA2 TN / 94-49

[0904]

[0905]

[0906]

[0907]

[0908]

[0909]

[0910]

[0911] While the invention has been particularly shown and described with reference to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made without departing from the scope of the invention as encompassed by the appended claims.

Claims

1. A compound represented by formula (I): or a pharmaceutically acceptable salt thereof, in: A is selected from: 1) optionally substituted aryl; and 2) optionally substituted heteroaryl; E is selected from: 1) optionally substituted aryl; and 2) optionally substituted heteroaryl; R 1 and R 2 Each independently selected from: 1) Hydrogen; 2) optionally substituted -C 1 -C 8 alkyl; 3) optionally substituted -C 3 -C 8 Cycloalkyl; 4) optionally substituted 3 to 8 membered heterocycloalkyl; 5) optionally substituted aryl; 6) optionally substituted aralkyl; 7) optionally substituted heteroaryl; and 8) optionally substituted heteroaralkyl; Or, R 1 and R 2 Together with the nitrogen atom to which they are attached, they form an optionally substituted 3- to 8-membered heterocyclic ring; R 3 is hydroxyl or fluorine; R 4 Selected from: 1) optionally substituted aryl; 2) optionally substituted aralkyl; 3) optionally substituted heteroaryl; and 4) Optionally substituted heteroaralkyl.

2. The compound according to claim 1, wherein R 4 Selected from the following groups: Where R 31 Halogen, -CN, -OR 33 , -CO 2 R 33 、-SO 2 R 33 、-SO 2 NR 33 R 34 、-NR 33 R 4 , optionally substituted -C 1 -C 6 Alkyl, optionally substituted -C 2 -C 6 alkenyl or optionally substituted -C 3 -C 8 Cycloalkyl; n is 0, 1 or 2; R 33 and R 34 are each independently selected from hydrogen, optionally substituted -C 1 -C 6 Alkyl, optionally substituted -C 2 -C 6 Alkenyl, optionally substituted -C 3 -C 8 cycloalkyl, optionally substituted 3 to 8 membered heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl.

3. A compound selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof:

4. A pharmaceutical composition comprising the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

5. A method for treating or preventing respiratory syncytial virus (RSV) infection in a subject in need thereof, comprising administering to the subject an effective therapeutic amount of a compound or combination of compounds as described in any one of claims 1 to 3.

6. The method of claim 5, further comprising the step of administering to the subject an additional anti-RSV drug.

7. The method of claim 5, further comprising administering to the subject a steroidal anti-inflammatory compound.

8. A method of treating respiratory syncytial virus infection and influenza in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1 to 3 and a therapeutically effective amount of an anti-influenza agent.

9. The method of claim 6, wherein the compound and the additional anti-RSV drug are co-formulated.

10. The method of claim 6, wherein the compound and the additional anti-RSV drug are co-administered.

11. The method of claim 6, wherein administration of the compound allows for administration of the additional anti-RSV drug at a lower dose or frequency than administration of the additional anti-RSV drug alone, which dose or frequency is necessary to achieve similar results in the prophylactic treatment of RSV infection in a subject in need thereof.

12. A method for treating or preventing HMPV infection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or combination of compounds as claimed in any one of claims 1 to 3.

13. The method of claim 12, further comprising the step of administering to the subject an additional anti-HMPV drug.

14. The method of claim 13, wherein the compound and the additional anti-HMPV drug are co-formulated.

15. The method of claim 13, wherein the compound and the additional anti-HMPV drug are co-administered.

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