RU2831561C2 - Compound, endothelin receptor antagonist and pharmaceutical composition - Google Patents

Abstract

FIELD: pharmaceutics.

SUBSTANCE: invention relates to a compound of formula (1), an endothelin A receptor antagonist, as well as a pharmaceutical composition based thereon. where R¹ and R² are all independently hydrogen or alkyl, R³ and R⁴ are all independently hydrogen or alkyl, or R³ and R⁴ together form an oxo group, R⁵-R⁷ are all independently hydrogen, alkyl, haloalkyl, halogen, alkoxy group or haloalkoxy group, each R⁸ independently denotes alkyl, haloalkyl or halogen, n is integer from 0 to 3, and Ar is described by the following formula (Ar1) or (Ar2): where X and Y denote nitrogen and oxygen, respectively, or oxygen and nitrogen, respectively, R⁹ and R¹⁰ independently denote hydrogen, alkyl, halogenalkyl or halogen, each R¹¹ independently denotes an alkyl, haloalkyl, halogen, alkoxy group or haloalkoxy group and m is integer from 0 to 3, where: alkyl represented by each of R¹, R², R³, R⁴, R⁵-R⁷, R⁸, R⁹, R¹⁰ and R¹¹, is an alkyl having from 1 to 6 carbon atoms, where the haloalkyl represented by each of R⁵-R⁷, R⁸, R⁹, R¹⁰ and R¹¹ is a haloalkyl having from 1 to 6 carbon atoms, where alkoxy represented by each of R⁵-R⁷ and R¹¹ is alkoxy having from 1 to 6 carbon atoms, where the haloalkoxy represented by each of R⁵-R⁷ and R¹¹ is a haloalkoxy having from 1 to 6 carbon atoms or a pharmaceutically acceptable salt thereof.

EFFECT: novel compounds which can be used for preventing or treating a disease associated with the endothelin A receptor are obtained.

15 cl, 7 tbl, 13 ex

Description

LEVEL OF TECHNOLOGY

FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES

[0001]

The present invention relates to a compound, an endothelin receptor A antagonist, and a pharmaceutical composition.

DESCRIPTION OF THE RELATED ART

[0002]

Endothelin (ET) is a peptide produced in vascular endothelial cells and has a vasoconstrictor effect. Endothelin receptor A (ETA) and endothelin receptor B (ETB) are known as endothelin receptors. The ETA receptor is involved in vasoconstriction, central activation of the sympathetic nervous system, stimulation of aldosterone secretion, etc. The ETB receptor is involved in vasodilation, endothelin clearance, etc.

[0003]

The ETA receptor has been reported to be associated with various diseases. For example, the ETA receptor has been reported to be associated with pulmonary hypertension (e.g., The New England Journal of Medicine, 2002, 346, pp 896–903; and International Heart Journal, 2020, 61, pp 799–805), focal segmental glomerulosclerosis (e.g., Journal of the American Society of Nephrology, 2018, 29, pp 2745–2754), IgA nephropathy (e.g., Nephron, 1996, 72, pp 454–460), chronic kidney disease (including diabetic nephropathy) (e.g., The Lancet, 2019, 393, pp 1937–1947; and Hypertension, 2011, 57, pp 772–779), and coronary heart disease (e.g., Bronchopulmonary disease, 2015, 28, pp 188–190). kidney disease associated with sickle cell syndrome (e.g., Journal of the American Society of Nephrology, 2017, 28, pp 2443–2458), acute kidney injury (e.g., Transplantation, 2001, 71, pp 211–216), hypertension (e.g., Hypertension, 2008, 52, pp 452–459), nonalcoholic steatohepatitis (NASH) (e.g., Drug Research, 2015, 65, pp 272–280; World Journal of Hepatology, 2016, 8, pp 933–941; and Digestive Diseases and Sciences, 2007, 52, pp 2622–2628), cancer (e.g., Nature reviews cancer, 2013, 13, pp 637–651), pain associated with endometriosis (e.g., European Journal of Pain, 2018, 22, pp 501–510; and Fertility and Sterility, 1994, 61, pp 1083–1087), complications associated with scleroderma (e.g., Inflammation & Allergy - Drug Targets, 2011, 10, pp 19–26; and Annals of the Rheumatic Diseases, 2011, 70, pp 32–38), cerebral vasospasm (e.g., Molecular medicine reports, 2018, 18, pp 5229–5236), and hypertrophic cardiomyopathy (e.g., Journal of the American Heart Association, 2014, 3, e00126).

[0004]

Various compounds have been described that have antagonist effects on the ETA receptor (e.g., U.S. Patent Nos. 5,514,696 and 5,612,359, and International Publication Nos. WO 1996/40681, WO 1996/31492, WO 1998/49162, WO 1998/13366, WO 1998/33780, WO 1998/33781, WO 2001/49685, WO 2004/35057, and WO 2013/115162). In particular, sitaxentan, which has an antagonistic effect on the ETA receptor, is known to selectively inhibit the ETA receptor over the ETB receptor and has been reported to have significant drug efficacy in clinical trials of pulmonary hypertension associated with connective tissue disease (European Heart Journal, 2008, 29, pp 1936-1948) without increasing blood endothelin concentrations (Pharmacological Reviews, 2016, 68, pp. 357-418) or causing peripheral edema (Annual Review of Pharmacology and Toxicology, 2007, 47, pp 731-759).

Related literature

[0005]

Patent Literature 1: U.S. Patent No. 5514696

Patent Literature 2: U.S. Patent No. 5612359

Patent Literature 3: WO 1996/40681

Patent Literature 4: WO 1996/31492

Patent Literature 5: WO 1998/49162

Patent Literature 6: WO 1998/13366

Patent Literature 7: WO 1998/33780

Patent Literature 8: WO 1998/33781

Patent Literature 9: WO 2001/49685

Patent Literature 10: WO 2004/35057

Patent Literature 11: WO 2013/115162

[0006]

Non-patent literature 1: The New England Journal of Medicine, 2002, 346, pp 896-903

Non-patent literature 2: International Heart Journal, 2020, 61, pp 799-805

Non-patent literature 3: Journal of the American Society of Nephrology, 2018, 29, pp 2745-2754

Non-patent literature 4: Nephron, 1996, 72, pp 454-460

Non-patent literature 5: The Lancet, 2019, 393, pp 1937-1947

Non-Patent Literature 6: Hypertension, 2011, 57, pp 772-779

Non-patent literature 7: Journal of the American Society of Nephrology, 2017, 28, pp 2443-2458

Non-patent literature 8: Transplantation, 2001, 71, pp 211-216

Non-Patent Literature 9: Hypertension, 2008, 52, pp 452-459

Non-Patent Literature 10: Drug Research, 2015, 65, pp 272-280

Non-patent literature 11: World Journal of Hepatology, 2016, 8, pp 933-941

Non-patent literature 12: Digestive Diseases and Sciences, 2007, 52, pp 2622-2628

Non-patent literature 13: Nature reviews cancer, 2013, 13, pp 637-651

Non-patent literature 14: European Journal of Pain, 2018, 22, pp 501-510

Non-Patent Literature 15: Fertility and Sterility, 1994, 61, pp 1083- 1087

Non-Patent Literature 16: Inflammation & Allergy - Drug Targets, 2011, 10, pp 19-26

Non-patent literature 17: Annals of the Rheumatic Diseases, 2011, 70, pp 32-38

Non-patent literature 18: Molecular medicine reports, 2018, 18, pp 5229-5236

Non-Patent Literature 19: Journal of the American Heart Association, 2014, 3, e00126

Non-patent literature 20: European Heart Journal, 2008, 29, pp 1936-1948

Non-patent literature 21: Pharmacological Reviews, 2016, 68, pp. 357-418

Non-patent literature 22: Annual Review of Pharmacology and Toxicology, 2007, 47, pp 731-759

[0007]

The object of the present invention is a compound that has an antagonistic effect on the ETA receptor, and an ETA receptor antagonist or a pharmaceutical composition containing the compound.

ESSENCE OF THE INVENTION

[0008]

The present inventors have conducted thorough studies and as a result completed the present invention by establishing that a compound having a special structure has an antagonistic effect on the ETA receptor.

[0009]

The present invention includes the following embodiments.

[1]

A compound described by the following formula (1):

(1)

Where

R ¹ and R ² all independently represent hydrogen or alkyl,

R ³ and R ⁴ are all independently hydrogen or alkyl, or

R ³ and R ⁴ together form an oxo group,

R ⁵ - R ⁷ all independently represent hydrogen, alkyl, haloalkyl, halogen, alkoxy or haloalkoxy,

each R ⁸ is independently alkyl, haloalkyl or halogen,

n is an integer between 0 and 3, and

Ar is described by the following formula (Ar1) or (Ar2):

Where

X and Y represent nitrogen and oxygen respectively or oxygen and nitrogen respectively,

R ⁹ and R ¹⁰ all independently represent hydrogen, alkyl, haloalkyl or halogen,

each R ¹¹ is independently alkyl, haloalkyl, halogen, alkoxy or haloalkoxy and

m is an integer between 0 and 3,

or a pharmaceutically acceptable salt thereof.

[2]

A compound or a pharmaceutically acceptable salt thereof according to paragraph [1], wherein each of R ¹ and R ² is hydrogen.

[3]

The compound or pharmaceutically acceptable salt thereof according to paragraph [1] or [2], wherein each of R ⁵ and R ⁷ is hydrogen and R ⁶ is alkyl.

[4]

A compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [3], where n is 0.

[5]

A compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [4], wherein Ar is described by the formula (Ar1).

[6]

A compound or pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [5], wherein each of R ⁹ and R ¹⁰ is alkyl.

[7]

A compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [6], wherein X and Y are nitrogen and oxygen, respectively.

[8]

A compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [6], wherein X and Y are oxygen and nitrogen, respectively.

[9]

A compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [4], wherein Ar is described by the formula (Ar2).

[10]

A compound or pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [4] and [9], wherein each R ¹¹ is independently alkyl or alkoxy.

[11]

A compound or pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [4], [9] and [10, where m is 2.

[12]

A compound or a pharmaceutically acceptable salt thereof according to paragraph [1], wherein the compound is selected from the group consisting of the following compounds:

[13]

An endothelin A receptor antagonist comprising a compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [12].

[14]

An endothelin receptor A antagonist according to paragraph [13], wherein the endothelin receptor A antagonist selectively inhibits endothelin receptor A over endothelin receptor B.

[15]

A pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [12].

[16]

A pharmaceutical composition according to paragraph [15] for the prevention or treatment of a disease selected from the group consisting of the following: pulmonary hypertension, nephropathy, hypertension, hepatitis, cancer, pain, complications associated with an autoimmune disease, heart failure and vasospasm.

[0010]

The present invention also includes the following embodiments.

[A1]

A method for inhibiting endothelin receptor A, comprising administering an effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [12] to a patient in need thereof.

[A2]

The method of paragraph [A1], wherein the method provides selective inhibition of endothelin receptor A over endothelin receptor B.

[A3]

A method for preventing or treating a disease comprising administering an effective amount of a compound or a pharmaceutically acceptable salt thereof according to any of paragraphs [1] to [12] to a patient in need thereof.

[A4]

The method of paragraph [A3], wherein the disease is selected from the group consisting of the following: pulmonary hypertension, nephropathy, hypertension, hepatitis, cancer, pain, complications associated with an autoimmune disease, heart failure, and vasospasm.

[0011]

[B1]

A compound or pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [12] for use in inhibiting endothelin receptor A.

[B2]

A compound or pharmaceutically acceptable salt thereof according to paragraph [B1], wherein its use provides selective inhibition of endothelin receptor A over endothelin receptor B.

[B3]

A compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [12] for use in the prevention or treatment of a disease.

[B4]

The compound or pharmaceutically acceptable salt thereof according to paragraph [B3], wherein the disease is selected from the group consisting of pulmonary hypertension, nephropathy, hypertension, hepatitis, cancer, pain, complications associated with an autoimmune disease, heart failure, and vasospasm.

[0012]

[C1]

The use of a compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [12] for inhibiting endothelin receptor A.

[C2]

Use according to paragraph [C1], where the use provides selective inhibition of endothelin receptor A as opposed to endothelin receptor B.

[C3]

The use of a compound or a pharmaceutically acceptable salt thereof according to any of paragraphs [1] to [12] for the prevention or treatment of a disease.

[C4]

The use according to paragraph [C3], where the disease is selected from the group consisting of the following: pulmonary hypertension, nephropathy, hypertension, hepatitis, cancer, pain, complications associated with autoimmune disease, heart failure and vasospasm.

[0013]

[D1]

The use of a compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs [1] to [12] for the preparation of an endothelin receptor A antagonist.

[D2]

Use according to paragraph [D1], wherein the endothelin receptor A antagonist selectively inhibits endothelin receptor A over endothelin receptor B.

[D3]

The use of a compound or a pharmaceutically acceptable salt thereof according to any of paragraphs [1] to [12] for the preparation of a pharmaceutical composition for the prevention or treatment of a disease.

[D4]

The use according to paragraph [D3], where the disease is selected from the group consisting of the following: pulmonary hypertension, nephropathy, hypertension, hepatitis, cancer, pain, complications associated with autoimmune disease, heart failure and vasospasm.

[0014]

The present invention can provide a compound having an antagonistic effect on an ETA receptor and an ETA receptor antagonist or a pharmaceutical composition containing the compound.

DETAILED DESCRIPTION OF PREFERRED IMPLEMENTATION OPTIONS

[0015]

Below, the present invention specifically describes embodiments of the present invention. However, the present invention is not limited thereto, and various changes or modifications can be made thereto without departing from the essence of the present invention.

[0016]

<Connection>

In one embodiment, the present invention relates to a compound described by the following formula (1):

(1)

Where

R ¹ and R ² all independently represent hydrogen or alkyl,

R ³ and R ⁴ are all independently hydrogen or alkyl, or

R ³ and R ⁴ together form an oxo group,

R ⁵ - R ⁷ all independently represent hydrogen, alkyl, haloalkyl, halogen, alkoxy or haloalkoxy,

each R ⁸ is independently alkyl, haloalkyl or halogen,

n is an integer between 0 and 3, and

Ar is described by the following formula (Ar1) or (Ar2):

Where

X and Y represent nitrogen and oxygen respectively or oxygen and nitrogen respectively,

R ⁹ and R ¹⁰ all independently represent hydrogen, alkyl, haloalkyl or halogen,

each R ¹¹ is independently alkyl, haloalkyl, halogen, alkoxy or haloalkoxy and

m is an integer between 0 and 3,

or a pharmaceutically acceptable salt thereof.

[0017]

The alkyl (including alkyl in haloalkyl; the same applies below in the present invention) used in the present invention may be linear, may be branched, or may be cyclic.

The alkyl moiety in an alkoxy group (including an alkoxy group in a haloalkoxy group; the same applies below in the present invention) used in the present invention may be linear, may be branched, or may be cyclic.

The alkyl and alkoxy groups used in the present invention may be substituted with a substituent or may be unsubstituted. Examples of the substituent include halogen, as well as hydroxy group, alkoxy group, amino group, carboxyl and carbonyl.

[0018]

In formula (1), R ¹ and R ² all independently represent hydrogen or alkyl, preferably hydrogen.

Alkyl represented by each R ¹ and R ² is preferably alkyl containing from 1 to 6 carbon atoms, more preferably alkyl containing from 1 to 3 carbon atoms, even more preferably methyl.

[0019]

In formula (1), R ³ and R ⁴ are all independently hydrogen or alkyl, or R ³ and R ⁴ together form an oxo group (=O). Preferably, R ³ and R ⁴ are all independently hydrogen or alkyl.

Alkyl represented by each R ³ and R ⁴ is preferably alkyl containing from 1 to 6 carbon atoms, more preferably alkyl containing from 1 to 3 carbon atoms, even more preferably methyl.

[0020]

In formula (1), R ⁵ represents hydrogen, alkyl, haloalkyl, halogen, alkoxy or haloalkoxy, preferably hydrogen.

Alkyl represented by R ⁵ is preferably alkyl containing from 1 to 6 carbon atoms, more preferably alkyl containing from 1 to 3 carbon atoms, even more preferably methyl.

The alkoxy group represented by R ⁵ is preferably an alkoxy group containing from 1 to 6 carbon atoms, more preferably an alkoxy group containing from 1 to 3 carbon atoms, even more preferably a methoxy group.

The halogen in the haloalkyl and haloalkoxy group represented by R ⁵ is preferably fluorine, chlorine, bromine or iodine, more preferably fluorine or chlorine.

[0021]

In formula (1), R ⁶ represents hydrogen, alkyl, haloalkyl, halogen, alkoxy or haloalkoxy, preferably alkyl.

Alkyl represented by R ⁶ is preferably alkyl containing from 1 to 6 carbon atoms, more preferably alkyl containing from 1 to 3 carbon atoms, even more preferably methyl.

The alkoxy group represented by R ⁶ is preferably an alkoxy group containing from 1 to 6 carbon atoms, more preferably an alkoxy group containing from 1 to 3 carbon atoms, even more preferably a methoxy group.

The halogen in the haloalkyl and haloalkoxy group represented by R ⁶ is preferably fluorine, chlorine, bromine or iodine, more preferably fluorine or chlorine.

[0022]

In formula (1), R ⁷ represents hydrogen, alkyl, haloalkyl, halogen, alkoxy or haloalkoxy, preferably hydrogen.

Alkyl represented by R ⁷ is preferably alkyl containing from 1 to 6 carbon atoms, more preferably alkyl containing from 1 to 3 carbon atoms, even more preferably methyl.

The alkoxy group represented by R ⁷ is preferably an alkoxy group containing 1 to 6 carbon atoms, more preferably an alkoxy group containing 1 to 3 carbon atoms, even more preferably a methoxy group.

The halogen in the haloalkyl and haloalkoxy group represented by R ⁷ is preferably fluorine, chlorine, bromine or iodine, more preferably fluorine or chlorine.

[0023]

In formula (1), each R ⁸ independently represents alkyl, haloalkyl or halogen, preferably alkyl or halogen.

Alkyl represented by R ⁸ is preferably alkyl containing from 1 to 6 carbon atoms, more preferably alkyl containing from 1 to 3 carbon atoms, even more preferably methyl.

The halogen in the haloalkyl represented by R ⁸ is preferably fluorine, chlorine, bromine or iodine, more preferably fluorine or chlorine.

[0024]

In formula (1), n is an integer from 0 to 3, preferably 0 or 1, more preferably 0.

[0025]

In formula (1), Ar is described by formula (Ar1) or (Ar2).

[0026]

In the formula (Ar1), X and Y represent nitrogen and oxygen respectively, or oxygen and nitrogen respectively.

[0027]

In the formula (Ar1), R ⁹ is hydrogen, alkyl, haloalkyl or halogen, preferably alkyl.

Alkyl represented by R ⁹ preferably means alkyl containing from 1 to 6 carbon atoms, more preferably alkyl containing from 1 to 3 carbon atoms, even more preferably methyl.

The halogen in the haloalkyl represented by R ⁹ is preferably fluorine, chlorine, bromine or iodine, more preferably fluorine or chlorine.

[0028]

In the formula (Ar1), R ¹⁰ is hydrogen, alkyl, haloalkyl or halogen, preferably alkyl.

Alkyl represented by R ¹⁰ is preferably alkyl containing from 1 to 6 carbon atoms, more preferably alkyl containing from 1 to 3 carbon atoms, even more preferably methyl.

The halogen in the haloalkyl represented by R ¹⁰ is preferably fluorine, chlorine, bromine or iodine, more preferably fluorine or chlorine.

[0029]

In formula (Ar2), each R ¹¹ independently represents alkyl, haloalkyl, halogen, alkoxy or haloalkoxy, preferably alkyl or alkoxy, more preferably alkyl and alkoxy.

Alkyl represented by R ¹¹ is preferably alkyl containing from 1 to 6 carbon atoms, more preferably alkyl containing from 1 to 3 carbon atoms, even more preferably methyl.

The alkoxy group represented by R ¹¹ is preferably an alkoxy group containing 1 to 6 carbon atoms, more preferably an alkoxy group containing 1 to 3 carbon atoms, even more preferably a methoxy group.

The halogen in the haloalkyl and haloalkoxy group represented by R ¹¹ is preferably fluorine, chlorine, bromine or iodine, more preferably fluorine or chlorine.

[0030]

In formula (Ar2), m is an integer between 0 and 3, preferably 2.

[0031]

The compound represented by the following formula (1) is not particularly limited and is preferably any of the following compounds.

[0032]

The pharmaceutically acceptable salt of the compound represented by formula (1) is not particularly limited as long as the salt can be used as a medicine. Examples thereof may include: inorganic acid salts such as hydrochloride, sulfate, nitrate, phosphate and hydrobromide; organic acid salts such as fumarate, maleate, malate, tartrate, succinate, citrate, methanesulfonate, p-toluenesulfonate, acetate, lactate and palmitate; alkali metal salts and alkaline earth metal salts.

[0033]

The compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof may form a solvate such as a hydrate. The solvate used in the present invention is included among the compounds represented by the formula (1) or pharmaceutically acceptable salts thereof.

[0034]

If a compound represented by formula (1) or a pharmaceutically acceptable salt thereof has stereoisomers (e.g., enantiomers and diastereoisomers), the individual stereoisomers and their mixture (e.g., a racemate) are included among the compounds represented by formula (1) or their pharmaceutically acceptable salts.

[0035]

The form of the compound represented by formula (1) or a pharmaceutically acceptable salt thereof is not particularly limited. The compound represented by formula (1) or a pharmaceutically acceptable salt thereof may be in the form of crystals or may be in an amorphous form. If the compound represented by formula (1) or a pharmaceutically acceptable salt thereof has crystalline polymorphs, the compound represented by formula (1) or a pharmaceutically acceptable salt thereof may be in any crystalline form.

[0036]

<Endothelin receptor A antagonist>

In one embodiment, the present invention relates to an ETA receptor antagonist comprising a compound described by formula (1) or a pharmaceutically acceptable salt thereof. The ETA receptor and the ETB receptor are known endothelin receptors. Inhibition of the ETB receptor can lead to problems such as an increase in endothelin levels in the blood or the occurrence of peripheral edema. Preferably, the ETA receptor antagonist of this embodiment can selectively inhibit endothelin receptor A over endothelin receptor B.

[0037]

In particular, the ratio (ETB receptor inhibition concentration (IC ₅₀ ))/(ETA receptor inhibition concentration (IC ₅₀ )) is preferably 2000 or more, more preferably 4000 or more, even more preferably 6000 or more, even more preferably 8000 or more, particularly preferably 10000 or more. The upper limit of the ratio (ETB receptor inhibition concentration (IC ₅₀ ))/(ETA receptor inhibition concentration (IC ₅₀ )) is not particularly limited and may be, for example, 100000, 80000, 60000 or 40000. The ETB receptor inhibition concentration and the ETA receptor inhibition concentration can be measured by the method described in the Examples.

[0038]

The ETA receptor inhibition concentration (IC ₅₀ ) of the ETA receptor antagonist of this embodiment is preferably 10 nM or less, more preferably 8.0 nM or less, still more preferably 6.0 nM or less, still more preferably 4.0 nM or less, particularly preferably 2.0 nM or less. The lower limit of the ETA receptor inhibition concentration (IC ₅₀ ) is not particularly limited and may be, for example, 0.005 nM, 0.01 nM or 0.05 nM.

[0039]

The ETB receptor inhibition concentration (IC ₅₀ ) of the ETA receptor antagonist of this embodiment is preferably 1000 nM or more, more preferably 2000 nM or more, even more preferably 3000 nM or more, still more preferably 4000 nM or more, particularly preferably 5000 nM or more. The upper limit of the ETB receptor inhibition concentration (IC ₅₀ ) is not particularly limited and may be, for example, 10000 nM, 15000 nM or 20000 nM.

[0040]

Use of the ETA receptor antagonist of this embodiment can treat and/or prevent a disease associated with (mediated by) the ETA receptor.

[0041]

<Pharmaceutical composition>

In one embodiment, the present invention relates to a pharmaceutical composition comprising a compound described by formula (1) or a pharmaceutically acceptable salt thereof.

[0042]

Examples of the disease targeted by the pharmaceutical composition of this embodiment may include pulmonary hypertension, nephropathy, hypertension, hepatitis, cancer, pain, complications associated with autoimmune disease, heart failure, and vasospasm.

Examples of nephropathy may include focal segmental glomerulosclerosis, IgA nephropathy, chronic kidney disease (including diabetic nephropathy), and kidney disease associated with sickle cell syndrome.

Examples of hepatitis may include nonalcoholic steatohepatitis (NASH) and alcoholic hepatitis.

Examples of cancer may include prostate cancer, squamous cell cancer, non-small cell lung cancer, and melanoma.

Examples of pain may include pain associated with endometriosis, chronic pain, neuropathic pain, cancer pain, and inflammatory pain.

Examples of complications associated with an autoimmune disease may include complications associated with scleroderma and complications associated with vasculitis.

Examples of heart failure may include right heart failure, left heart failure, heart failure associated with hypertrophic cardiomyopathy, heart failure associated with dilated cardiomyopathy, heart failure with preserved ejection fraction, and heart failure with reduced ejection fraction.

Examples of vasospasm may include cerebral vasospasm and vasospastic angina.

[0043]

The pharmaceutical composition of this embodiment can be administered orally or parenterally. Examples of the dosage form for oral administration include tablets, pills, granules, powders, capsules, syrups, emulsions and suspensions. Examples of the dosage form for parenteral administration include injections, infusions, drops, eye drops and suppositories.

[0044]

The pharmaceutical composition of this embodiment may optionally contain an inert filler, a binder, a lubricant, a disintegrant, a sweetener, a surfactant, a suspending agent, an emulsifying agent, a coloring agent, a preservative, a flavoring agent, a drug modifier, a stabilizer, a thickener, etc.

[0045]

The dose of the pharmaceutical composition of this embodiment varies depending on the condition or body weight of the patient, the type of compound, the type of disease, the route of administration, etc., and the appropriate amount can be determined by a physician.

[0046]

<Methodology for obtaining the compound>

The compound represented by formula (1) or a pharmaceutically acceptable salt thereof can be synthesized by appropriately applying a method known in the art. Examples of the synthesis method may include the following schemes A - C:

<Diagram A>

<Scheme B>

<Diagram C>

[0047]

In schemes A - C, R ¹ - R ⁸ , n and Ar are as defined above, Z ¹ and Z ² each denote halogen, and Pro denotes a protecting group.

[0048]

In Scheme A, compound (1) is reacted with, for example, sodium hydride to yield compound (2) (step 1). Compound (2) is reacted with a halogen (for example, iodine) to yield compound (3) (step 2).

[0049]

In Scheme B, the amino group of compound (4) is protected with a protecting group (e.g., methoxymethyl group) to give compound (5) (step 3). Compound (5) is reacted with triisopropylsilylacetylene to give compound (6) (step 4). Compound (6) is reacted with, e.g., tetrabutylammonium fluoride to give compound (7) (step 5).

[0050]

In Scheme C, compound (3) is reacted with compound (7) to yield compound (8) (step 6). The protecting group is removed from compound (8) to yield compound (9) (step 7).

[0051]

If the compound represented by formula (1) or a pharmaceutically acceptable salt thereof has stereoisomers, the isomers can be separated by a technique known in the art. Examples of a technique known in the art may include chromatography, enzymatic techniques, and crystallization techniques.

[Examples]

[0052]

Below, the present invention is described in more detail with reference to examples and comparative examples. However, the technical scope of the present invention is not limited to them.

[0053]

[Example of receiving 1-1]

6-Bromo-5-methylisobenzofuran-1(3H)-one

To a mixture of 3-bromo-4-methylbenzoic acid (1.5 g, 7.01 mmol) and 1,1-dibromomethane (20 ml) were added potassium hydrogen phosphate (3.6 g, 21.0 mmol) and palladium(II) acetate (157 mg, 0.70 mmol) at room temperature, and the reaction mixture was stirred at 140°C for 48 h. The reaction mixture was returned to room temperature and filtered through celite, and washed with ethyl acetate. The solvent in the filtrate was distilled off under reduced pressure, and the residue was triturated with ethanol to give the title compound (1.3 g).

1H NMR (400 MHz, CDCl3): δ 8.09 (s, 1H), 7.37 (s, 1H), 5.23 (s, 2H), 2.53 (s, 3H).

[0054]

[Example of receiving 1-2]

6-Bromo-1,1,5-trimethyl-1,3-dihydroisobenzofuran

To a mixture of 6-bromo-5-methylisobenzofuran-1(3H)-one (1.00 g, 4.41 mmol) and tetrahydrofuran (10 ml) was added methyl magnesium iodide (3.0 M solution in diethyl ether, 4.5 ml, 13.5 mmol) at -15°C, and the mixture was stirred at the same temperature as above for 2 h. The reaction mixture was added ice-cooled water, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (615 mg).

1H NMR (400 MHz, CDCl3): δ 7.45 (d, J=5.2 Hz, 1H), 7.20 (d, J=5.2 Hz, 1H), 4.77 (d, J= 5.2 Hz, 2H), 2.37 (d, J=5.2 Hz, 3H), 1.68 (d, J=5.2 Hz, 6H).

[0055]

[Example of receiving 1-3]

2-Chloro-N-(4,5-dimethylisoxazol-3-yl)pyridin-3-sulfonamide

To a mixture of 4,5-dimethylisoxazol-3-amine (1.0 g, 8.92 mmol) and pyridine (20.0 mL) were gradually added 4-dimethylaminopyridine (109 mg, 0.892 mmol) and 2-chloropyridine-3-sulfonyl chloride (2.84 g, 13.4 mmol) at 0 °C, and the reaction mixture was stirred at 50 °C for 16 h. The organic layer was washed with saturated brine and dried over sodium sulfate. The solvent was distilled off under reduced pressure to give the title compound (1.50 g) as a crude compound. This compound was used in the next reaction without further purification.

ESI-MS: m/z 288.0 [M+1]+

[0056]

[Example of receiving 1-4]

2-Chloro-N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)pyridine-3-sulfonamide

To a mixture of 2-chloro-N-(4,5-dimethylisoxazol-3-yl)pyridine-3-sulfonamide (1.80 g, 6.26 mmol) and DMF (30.0 ml) was gradually added 60% sodium hydride (695 mg, 17.4 mmol) at 0°C, and the mixture was stirred at the same temperature as above for 30 min. To the reaction mixture was gradually added chloromethyl methyl ether (1.06 ml, 13.9 mmol), and the mixture was stirred at room temperature for 3 h. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (1.20 g).

ESI-MS: m/z 332.07 [M+1]+

[0057]

[Example of receiving 1-5]

N-(4,5-Dimethylisoxazol-3-yl)-N-(methoxymethyl)-2-((triisopropylsilyl)ethynyl)pyridine-3-sulfonamide

Under argon atmosphere, bis(triphenylphosphine)palladium(II) dichloride (84.6 mg, 0.121 mmol) and copper(I) iodide (11.5 mg, 0.0362 mmol) were added to a mixture of 2-chloro-N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)pyridine-3-sulfonamide (400 mg, 1.21 mmol), triethylamine (0.50 mL, 3.62 mmol), triisopropylsilylacetylene (440 mg, 2.41 mmol) and THF (5.00 mL), and the reaction mixture was stirred at 55 °C for 6 h. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.150 g).

1H NMR (400 MHz, DMSO-d ₆ ) δ 8.82 (d, 1H, J=4.8 Hz), 8.21 (d, 1H, J=8 Hz), 7.63-7.60 ( m, 1H), 5.16 (s, 2H), 3.34 (s, 3H), 2.32 (s, 3H), 1.85 (s, 3H), 1.14 (bs, 21H).

[0058]

[Example of receiving 1-6]

N-(4,5-Dimethylisoxazol-3-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide

To a mixture of N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)-2-((triisopropylsilyl)ethynyl)pyridine-3-sulfonamide (150 mg, 4.3 mmol) and THF (3.00 mL) was gradually added tetrabutylammonium fluoride (1 mol/L tetrahydrofuran solution, 0.942 mL, 12.9 mmol) at 0°C, and the reaction mixture was stirred at room temperature for 2 h. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.060 g).

ESI-MS: m/z 322.19 [M+1]+

[0059]

[Example of obtaining 1-7]

N-(4,5-Dimethylisoxazol-3-yl)-N-(methoxymethyl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide

Under argon atmosphere, 6-bromo-1,1,5-trimethyl-1,3-dihydroisobenzofuran (0.36 g, 1.49 mmol), copper(II) sulfate (7.8 mg, 0.04 mmol), triethylamine (1.04 mL, 7.48 mmol), sodium ascorbate (24 mg, 0.12 mmol) and tetrakis(triphenylphosphine)palladium(0) (28 mg, 0.02 mmol) were added to a mixture of N-(4,5-dimethylisoxazol-3-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide (0.4 g, 1.25 mmol) and N,N-dimethylformamide (5 mL) and the reaction mixture was stirred at 80 °C for 4 h. The reaction mixture was returned to room temperature and Ice-cooled water was added, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high-performance liquid chromatography (Kinetex F5, (250×50 mm) 5 μm, mixed solution of 0.1% aqueous trifluoroacetic acid/acetonitrile-methanol (1:1)) to obtain the title compound (615 mg).

ESI-MS: m/z 482.29 [M+1]+

[0060]

[Example 1]

N-(4,5-Dimethylisoxazol-3-yl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide

To a mixture of N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide (90 mg, 0.18 mmol) and methanol (5 ml) was added 6 mol/L hydrochloric acid (1 ml) at 0°C, and the reaction mixture was stirred at 50°C for 4 h. The reaction mixture was returned to room temperature, and ice-cooled water was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high performance liquid chromatography (Omega Ps C18 (250×21.2 mm) 5 μm, mixed solution of 0.1% aqueous trifluoroacetic acid/acetonitrile-methanol (1:1)) to give the title compound (30 mg).

ESI-MS: m/z 438.24 [M+1]+

1H NMR (400 MHz, DMSO-d6): δ 11.06 (s, 1H), 8.83 (d, J=4.8 Hz, 2H), 8.35 (d, J=8 Hz, 1H) , 7.66-7.63 (m, 1H), 7.48 (s, 1H), 7.24 (s, 1H), 4.96 (s, 2H), 2.46 (s, 3H), 2.07 (s, 3H), 1.81 (s, 3H), 1.43 (s, 6H).

[0061]

[Example of Receipt 2-1]

(E)-N'-(2-Hydroxy-5-methylbenzylidene)acetohydrazide

To a mixture of 2-hydroxy-5-methylbenzaldehyde (10 g, 73.5 mmol) and ethanol (50 ml) was gradually added acetohydrazide (5.44 g, 73.5 mmol) at room temperature, and the reaction mixture was stirred at 90°C for 16 h. The solvent in the reaction mixture was distilled off under reduced pressure, and the residue was washed with hexane to give the title compound (10 g) as a crude compound. This compound was used in the next reaction without further purification.

[0062]

[Example of receiving 2-2]

2-Acetyl-5-methylbenzaldehyde

To a mixture of (E)-N'-(2-hydroxy-5-methylbenzylidene)acetohydrazide (6 g, 31.3 mmol) and tetrahydrofuran (90 mL) was added lead(IV) acetate (15.1 g, 34.4 mmol) at 0°C, and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was filtered through celite and washed with ethyl acetate. The filtrate was washed with saturated aqueous sodium bicarbonate and saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (1.3 g).

ESI-MS: m/z 163.1 [M+1]+

[0063]

[Example of receiving 2-3]

1-(2-(Hydroxymethyl)-4-methylphenyl)ethan-1-ol

To a mixture of 2-acetyl-5-methylbenzaldehyde (2.5 g, 15.4 mmol), tetrahydrofuran (10 mL) and ethanol (30 mL), sodium borohydride (2.9 g, 77.2 mmol) was added at 0°C, and the reaction mixture was stirred at 70°C for 2 h. The reaction temperature was returned to room temperature, and ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium bicarbonate and saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (1.8 g).

1H NMR (400 MHz, CDCl3): δ 7.38-7.35 (m, 1H), 7.16 (d, J=2.4 Hz, 2H), 5.19-5.14 (m, 1H ), 4.81 (t, J=8 Hz, 1H), 4.63 (d, J=12 Hz, 2H), 2.68 (s, 2H), 2.35 (d, J=4.8 Hz, 3H), 2.45 (d, J=5.2 Hz, 2H).

[0064]

[Example of receiving 2-4]

1,5-Dimethyl-1,3-dihydroisobenzofuran

To a mixture of 1-(2-(hydroxymethyl)-4-methylphenyl)ethan-1-ol (1.6 g, 9.64 mmol) and tetrahydrofuran (16 ml) was gradually added 60% sodium hydride (1.07 g, 11.2 mmol) at 0°C, and the mixture was stirred at the same temperature as above for 30 min. To the reaction mixture was added trimethyl phosphate (3.5 g, 24.1 mmol) at room temperature, and the mixture was stirred at the same temperature as above for 16 h. Water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (1.0 g).

1H NMR (400 MHz, CDCl3): δ 7.12-7.06 (m, 3H), 5.31 (d, J=6.4 Hz, 1H), 5.14-5.01 (m, 2H ), 2.39 (s, 3H), 1.50 (d, J=6.4 Hz, 3H).

[0065]

[Example of receiving 2-5]

6-Bromo-1,5-dimethyl-1,3-dihydroisobenzofuran

To a mixture of 1,5-dimethyl-1,3-dihydroisobenzofuran (700 mg, 4.73 mmol) and dichloromethane (7 ml) were added N-bromosuccinimide (842 mg, 4.73 mmol) and trifluoromethanesulfonic acid (700 mg, 4.73 mmol) at 0°C, and the mixture was stirred at room temperature for 2 h. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.7 g).

1H NMR (400 MHz, CDCl3): δ 7.32 (dd, J=8.4 Hz, J=9.2 Hz, 1H), 7.09 (d, J=8 Hz, 1H), 5.29 -5.24 (m, 1H), 5.07-5.02 (m, 1H), 4.95 (t, J=7.6 Hz, 1H), 2.41 (d, J=8.4 Hz, 3H), 1.49-1.46 (m, 3H).

[0066]

[Example of receiving 2-6]

2-((3,6-Dimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)-N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)pyridin-3-sulfonamide

Under argon atmosphere, 6-bromo-1,5-dimethyl-1,3-dihydroisobenzofuran (0.54 g, 2.40 mmol), copper(I) iodide (83 mg, 0.436 mmol), triethylamine (0.6 mL, 4.36 mmol) and tetrakis(triphenylphosphine)palladium(0) (250 mg, 0.218 mmol) were added to a mixture of N-(4,5-dimethylisoxazol-3-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide (0.7 g, 2.18 mmol) and N,N-dimethylformamide (5 mL), and the reaction mixture was stirred at 80 °C for 4 h. The reaction mixture was returned to room temperature and ice-cooled water was added, followed by extraction ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high performance liquid chromatography (Luna C8 (250×50 mm) 10 μm, 0.1% aqueous formic acid/acetonitrile) to give the title compound (50 mg).

ESI-MS: m/z 468.27 [M+1]+

[0067]

[Example 2]

2-((3,6-Dimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)-N-(4,5-dimethylisoxazol-3-yl)pyridin-3-sulfonamide

To a mixture of 2-((3,6-dimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)-N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)pyridine-3-sulfonamide (50 mg, 0.107 mmol) and methanol (5 ml) was added 6 mol/L hydrochloric acid (1 ml) at 0°C, and the reaction mixture was stirred at 50°C for 4 h. The reaction mixture was returned to room temperature, and ice-cooled water was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high performance liquid chromatography (Luna C8 (250×50 mm) 10 μm, 0.1% aqueous formic acid/acetonitrile) to afford the title compound (22 mg).

ESI-MS: m/z 424.31 [M+1]+

1H NMR (400 MHz, DMSO-d6): δ 11.06 (s, 1H), 8.82 (d, J=4 Hz, 1H), 8.34 (t, J=6.8 Hz, 1H) , 7.66-7.62 (m, 1H), 7.47 (d, J=5.2 Hz, 1H), 7.27 (d, J=5.2 Hz, 1H), 5.22 (d, J=9.6 Hz, 1H), 5.05-5.00 (m,1H), 4.95-4.90 ( m, 1H), 2.47 (s, 3H), 2.09 (d, J=4.8 Hz, 3H), 1.81 (d, J=4.8 Hz, 3H), 1.42 (t, J=5.2 Hz, 3H).

[0068]

[Example of receiving 3-1]

(4-Methyl-1,2-phenylene)dimethanol

To a mixture of 5-methylisobenzofuran-1,3-dione (100 g, 617 mmol) and THF (1000 mL) was gradually added borane dimethyl sulfide complex (156 mL, 1.85 mol) at 0 °C, and the reaction mixture was stirred at 80 °C for 16 h. The temperature of the reaction mixture was returned to room temperature, and methanol was added to the reaction mixture. The solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (40.0 g).

1H NMR (400 MHz, DMSO-d ₆ ) δ 7.23 (d, 1H, J=8.0 Hz), 7.18 (br s, 1H), 7.02 (d, 1H, J=7, 6 Hz), 5.02 (t, 1H, J=5.6 Hz), 4.98 (t, 1H, J=5.2 Hz), 4.49 (dd, 4H, J=8.4, 14 Hz), 2.27 (s, 3H).

[0069]

[Example of receiving 3-2]

5-Methyl-1,3-dihydroisobenzofuran

To a mixture of (4-methyl-1,2-phenylene)dimethanol (40.0 g, 263 mmol) and DMF (200 mL) was gradually added 50% sodium hydride (15.8 g, 657 mmol) at 0°C, and the mixture was stirred at the same temperature as above for 30 min. Trimethyl phosphate (92.0 g, 657 mmol) was gradually added to the reaction mixture, and the mixture was stirred at room temperature for 3 h. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (20.50 g).

1H NMR (400 MHz, CDCl ₃ ) δ 7.12 (d, 1H, J=7.6 Hz), 7.08 (br s, 1H), 7.06 (d, 1H, J=4.8 Hz ), 5.08 (s, 4H), 2.37 (s, 3H).

[0070]

[Example of getting 3-3]

5-Iodo-6-methyl-1,3-dihydroisobenzofuran

To a mixture of 5-methyl-1,3-dihydroisobenzofuran (1.00 g, 7.45 mmol), methanol (5.00 mL) and ethanol (5.00 mL) were gradually added silver sulfate (4.65 g, 14.9 mmol) and iodine (1.89 g, 14.9 mmol) at 0°C, and the mixture was stirred at the same temperature as above for 1 h. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.60 g).

1H NMR (400 MHz, CDCl ₃ ) δ 7.68 (s, 1H), 7.11 (s, 1H), 5.03 (d, 4H, J=5.2 Hz), 2.44 (s, 3H).

[0071]

[Example of receiving 3-4]

N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)-2-((6-methyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide

Under argon atmosphere, bis(triphenylphosphine)palladium(II) dichloride (899 mg, 0.778 mmol) and copper(I) iodide (148 mg, 0.778 mmol) were added to a mixture of N-(4,5-dimethylisoxazol-3-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide (2.50 g, 7.78 mmol), 5-iodo-6-methyl-1,3-dihydroisobenzofuran (2.23 g, 8.56 mmol), triethylamine (2.19 mL, 15.6 mmol) and toluene (30.0 mL), and the reaction mixture was stirred at 65°C for 6 h. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (1.60 g).

ESI-MS: m/z 454.27 [M+1]+

[0072]

[Example 3]

N-(4,5-Dimethylisoxazol-3-yl)-2-((6-methyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridin-3-sulfonamide

To a mixture of N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)-2-((6-methyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide (1.00 g, 2.21 mmol) and methanol (10.0 ml) was added 6 mol/L hydrochloric acid (5.0 ml) at room temperature, and the reaction mixture was stirred at 60°C for 16 h. The temperature of the reaction mixture was returned to room temperature, and the solvent was distilled off under reduced pressure. Ethyl acetate was added to the residue. The organic layer was washed with water and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.110 g).

1H NMR (400 MHz, DMSO-d ₆ ) δ 11.03 (s, 1H), 8.82 (dd, 1H, J=4.8, 1.6 Hz), 8.33 (dd, 1H, J =8.4, 1.6 Hz), 7.63-7.60 (m, 1H), 7.50 (bs, 1H), 7.29 (bs, 1H), 5.00 (s, 4H), 2.50 (s, 3H), 2.10 (s, 3H), 1.80 (s, 3H).

[0073]

[Example of obtaining 4-1]

N-(4,5-Dimethylisoxazol-3-yl)-N-(methoxymethyl)-2-((6-methyl-3-oxo-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide

Under argon atmosphere, 6-bromo-5-methylisobenzofuran-1(3H)-one (0.366 g, 1.61 mmol), copper(I) iodide (47 mg, 0.24 mmol), triethylamine (0.35 mL, 2.48 mmol) and tetrakis(triphenylphosphine)palladium(0) (143 mg, 0.12 mmol) were added to a mixture of N-(4,5-dimethylisoxazol-3-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide (0.4 g, 1.24 mmol) and N,N-dimethylformamide (5 mL), and the reaction mixture was stirred at 80 °C for 4 h. The reaction mixture was returned to room temperature and ice-cooled water was added thereto, followed by extraction ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high performance liquid chromatography (Luna C8 (250×50 mm) 10 μm, 0.1% aqueous formic acid/acetonitrile) to give the title compound (50 mg).

ESI-MS: m/z 468.24 [M+1]+

[0074]

[Example 4]

N-(4,5-Dimethylisoxazol-3-yl)-2-((6-methyl-3-oxo-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide

To a mixture of N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)-2-((6-methyl-3-oxo-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide (50 mg, 0.10 mmol) and methanol (5 ml) was added 6 mol/L hydrochloric acid (1 ml) at 0°C, and the reaction mixture was stirred at 50°C for 4 h. The reaction mixture was returned to room temperature, and ice-cooled water was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high performance liquid chromatography (Luna C18 (250×50 mm) 10 μm, 0.1% aqueous formic acid/acetonitrile) to afford the title compound (12 mg).

ESI-MS: m/z 424.18 [M+1]+

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.21 (s, 1H), 8.86 (t, J=5.2 Hz, 1H), 8.37 (t, J=5.2 Hz , 1H), 8.08 (d, J=4.8 Hz, 1H), 7.67 (t, J=4.8 Hz, 2H), 5.45 (d, J=5.2 Hz, 2H), 2.61 (d, J=5.2 Hz, 3H), 2.11 (d, J=5.2 Hz, 3H), 1, 81 (d, J=5.2 Hz, 3H).

[0075]

[Example of obtaining 5-1]

2-((1,3-Dihydroisobenzofuran-5-yl)ethynyl)-N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)pyridin-3-sulfonamide

Under argon atmosphere, 5-bromo-1,3-dihydroisobenzofuran (170 mg, 0.856 mmol), copper(I) iodide (15 mg, 0.077 mmol), triethylamine (0.22 mL, 1.56 mmol) and tetrakis(triphenylphosphine)palladium(0) (90 mg, 0.077 mmol) were added to a mixture of N-(4,5-dimethylisoxazol-3-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide (250 mg, 0.778 mmol) and toluene (10 mL), and the reaction mixture was stirred at 80 °C for 4 h. The reaction mixture was returned to room temperature and ice-cooled water was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high performance liquid chromatography (Luna C18 (250×50 mm) 10 μm, 0.1% aqueous formic acid/acetonitrile) to give the title compound (50 mg).

ESI-MS: m/z 440.27 [M+1]+

[0076]

[Example 5]

2-((1,3-Dihydroisobenzofuran-5-yl)ethynyl)-N-(4,5-dimethylisoxazol-3-yl)pyridin-3-sulfonamide

To a mixture of 2-((1,3-dihydroisobenzofuran-5-yl)ethynyl)-N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)pyridine-3-sulfonamide (100 mg, 0.227 mmol) and methanol (5 ml) was added 6 mol/L hydrochloric acid (1 ml) at 0°C, and the reaction mixture was stirred at 50°C for 8 h. The reaction mixture was returned to room temperature, and ice-cooled water was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated aqueous sodium bicarbonate and saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high performance liquid chromatography (Luna C18 (250×50 mm) 10 μm, 0.1% aqueous formic acid/acetonitrile) to afford the title compound (16 mg).

ESI-MS: m/z 396.23 [M+1]+

1H NMR (400 MHz, DMSO-d6): δ 11.10 (s, 1H), 8.80 (s, 1H), 8.34 (t, J=6.4 Hz, 1H), 7.63 ( t, J=13.2 Hz, 1H), 7.57-7.53 (m, 2H), 7.44 (t, J=6.4 Hz, 1H), 5.05 (d, J=8 Hz, 1H), 3.33 (s, 3H), 2.09 (s, 3H), 1.80 (s, 3H) .

[0077]

[Example of obtaining 6-1]

5-Chloro-6-iodoisobenzofuran-1(3H)-one

Under argon atmosphere, to a mixture of 4-chloro-3-iodobenzoic acid (1.0 g, 3.5 mmol), 1,1-dibromomethane (15 mL) and potassium hydrogen phosphate (2.25 g, 10.6 mmol) was added palladium(II) acetate (80 mg, 0.35 mmol), and the reaction mixture was stirred at 140°C for 16 h. The reaction mixture was returned to room temperature and filtered through celite and washed with ethyl acetate. The filtrate was washed with water and saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.4 g).

1H NMR (400 MHz, DMSO-d6): δ 8.35 (s, 1H), 7.97 (s, 1H), 5.34 (s, 2H).

[0078]

[Example of obtaining 6-2]

5-Chloro-6-iodo-1,3-dihydroisobenzofuran-1-ol

To a mixture of 5-chloro-6-iodoisobenzofuran-1(3H)-one (200 mg, 0.680 mmol) and dichloromethane (30 ml) was added diisobutylaluminum hydride (20% solution in toluene, 1.7 ml, 2.0 mmol) at -78°C, and the mixture was stirred at the same temperature as above for 2 h. Saturated aqueous ammonium chloride solution was added to the reaction mixture, and it was filtered through celite, and washed with ethyl acetate. The filtrate was washed with water and saturated brine, and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (70 mg).

1H NMR (400 MHz, DMSO-d6): δ 7.89 (s, 1H), 7.61 (s,1H),6.89 (d, J=7.6 Hz, 1H), 6.75 ( dd, J=7.6, 8.0 Hz, 1H),4.99 (d, J=13.6 Hz, 1H), 4.84 (d, J=12.8 Hz, 1H), 4.83 (d, J=13.6 Hz, 1H).

[0079]

[Example of obtaining 6-3]

5-Chloro-6-iodo-1,3-dihydroisobenzofuran

To a mixture of 5-chloro-6-iodo-1,3-dihydroisobenzofuran-1-ol (300 mg, 1.01 mmol) and dichloromethane (26 ml) were added trifluoroacetic acid (0.11 ml, 1.5 mmol) and triethylsilane (0.4 ml, 2.6 mmol) at room temperature, and the mixture was stirred at the same temperature as above for 4 h. The reaction mixture was added saturated aqueous sodium bicarbonate solution, which was then extracted with dichloromethane. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (200 mg).

1H NMR (400 MHz, DMSO-d6): δ 7.90 (s, 1H), 7.56 (s, 1H), 4.94 (s, 4H).

[0080]

[Example of getting 6-4]

2-((6-Chloro-1,3-dihydroisobenzofuran-5-yl)ethynyl)-N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)pyridine-3-sulfonamide

Under argon atmosphere, 5-chloro-6-iodo-1,3-dihydroisobenzofuran (126 mg, 0.448 mmol), copper(I) iodide (7.08 mg, 0.037 mmol), triethylamine (0.16 mL, 1.12 mmol) and tetrakis(triphenylphosphine)palladium(0) (43 mg, 0.037 mmol) were added to a mixture of N-(4,5-dimethylisoxazol-3-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide (120 mg, 0.373 mmol) and N,N-dimethylformamide (5 mL), and the reaction mixture was stirred at 60 °C for 2 h. The reaction mixture was returned to room temperature and ice-cooled water was added thereto, followed by extraction ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (95 mg).

ESI-MS: m/z 473.9 [M+1]+

[0081]

[Example 6]

2-((6-Chloro-1,3-dihydroisobenzofuran-5-yl)ethynyl)-N-(4,5-dimethylisoxazol-3-yl)pyridin-3-sulfonamide

To a mixture of 2-((6-chloro-1,3-dihydroisobenzofuran-5-yl)ethynyl)-N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)pyridine-3-sulfonamide (120 mg, 0.253 mmol), methanol (5 mL) and acetonitrile (1 mL) was added 50% sulfuric acid (1.2 mL) at 0°C, and the reaction mixture was stirred at 60°C for 2 h. The reaction mixture was returned to room temperature, added with saturated aqueous sodium bicarbonate solution, and then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high performance liquid chromatography (Kinetex penta C18 (250×50 mm) 5 μm, 0.1% aqueous trifluoroacetic acid/acetonitrile) to afford the title compound (40 mg).

ESI-MS: m/z 430.01 [M+1]+

1H NMR (400 MHz, DMSO-d6): δ 11.08 (s, 1H), 8.84 (d, J=4.0 Hz, 1H), 8.34 (d, J= 8.4 Hz, 1H), 7.69-7.65 (m, 1H), 7.64 (s,1H),7.61 (s,1H), 4.14 (s, 4H), 2.11 (s, 3H), 1.79 (s, 3H).

[0082]

[Example of obtaining 7-1]

2-(2-(Hydroxymethyl)-4-methylphenyl)propan-2-ol

To a mixture of 5-methylisobenzofuran-1(3H)-one (2.40 g, 16.2 mmol) and tetrahydrofuran (30 mL) was added methylmagnesium bromide (3.0 M solution in ether, 16.2 mL, 48.6 mmol) at 0°C, and the mixture was stirred at room temperature for 3 h. The reaction mixture was added saturated aqueous ammonium chloride solution, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (1.30 g).

1H NMR (400Hz, DMSO-d6) δ 7.30 (s, 1H), 7.19 (d, J=8.0 Hz, 1H), 6.95 (d, J=8.0 Hz, 1H) , 5.09 (s, 1H), 4.98 (t, J=5.6 Hz, 1H), 4.75 (d, J=5.6 Hz, 2H), 2.25 (s, 3H), 1.46 (s, 6H).

[0083]

[Example of obtaining 7-2]

1,1,5-Trimethyl-1,3-dihydroisobenzofuran

To a mixture of 2-(2-(hydroxymethyl)-4-methylphenyl)propan-2-ol (1.30 g, 7.22 mmol) and tetrahydrofuran (20 ml) was gradually added 50% sodium hydride (0.69 g, 14.4 mmol) at 0°C, and the mixture was stirred at room temperature for 1 h. To the reaction mixture was gradually added trimethyl phosphate (2.01 g, 14.4 mmol) at room temperature, and the mixture was stirred at the same temperature as above for 4 h. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.60 g).

1H NMR (400Hz, DMSO-d6) δ 7.11 (d, J=7.6 Hz, 1H), 7.04 (t, J=4.8 Hz, 2H), 4.89 (s, 2H) , 2.29 (s, 3H), 1.37 (s, 6H).

[0084]

[Example of getting 7-3]

6-Iodo-1,1,5-trimethyl-1,3-dihydroisobenzofuran

To a mixture of 1,1,5-trimethyl-1,3-dihydroisobenzofuran (0.60 g, 3.7 mmol) and dichloromethane (10 ml) were gradually added N-iodosuccinimide (1.60 g, 7.40 mmol) and trifluoromethanesulfonic acid (1.10 g, 7.40 mmol) at 0°C, and the mixture was stirred at the same temperature as above for 2 h. The reaction mixture was added 10% aqueous sodium thiosulfate solution, which was then extracted with dichloromethane. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.30 g).

1H NMR (400Hz, DMSO-d6) δ 7.71 (s, 1H), 7.22 (s, 1H), 4.86 (s, 2H), 2.35 (s, 3H), 1.38 ( s, 6H).

[0085]

[Example of getting 7-4]

2-Chloro-N-(4-chloro-5-methylisoxazol-3-yl)pyridine-3-sulfonamide

To a mixture of 4-chloro-5-methylisoxazol-3-amine (2.98 g, 22.6 mmol) and pyridine (50 ml) were gradually added 4-dimethylaminopyridine (0.69 g, 5.66 mmol) and 2-chloropyridine-3-sulfonyl chloride (4.0 g, 18.8 mmol) at 0 °C, and the reaction mixture was stirred at 55 °C for 16 h. The organic layer was washed with 2 N hydrochloric acid and saturated brine, and dried over sodium sulfate. The solvent was distilled off under reduced pressure to give the title compound (1.90 g) as a crude compound. This compound was used in the next reaction without further purification.

ESI-MS: m/z 307.98 [M+1]+

[0086]

[Example of getting 7-5]

2-Chloro-N-(4-chloro-5-methylisoxazol-3-yl)-N-(methoxymethyl)pyridine-3-sulfonamide

To a mixture of 2-chloro-N-(4-chloro-5-methylisoxazol-3-yl)pyridine-3-sulfonamide (1.90 g, 6.98 mmol) and N,N-dimethylformamide (20.0 ml) was gradually added 50% sodium hydride (594 mg, 12.3 mmol) at 0°C, and the mixture was stirred at the same temperature as above for 30 min. To the reaction mixture was gradually added chloromethyl methyl ether (0.69 ml, 9.28 mmol), and the mixture was stirred at room temperature for 3 h. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (1.0 g).

1H NMR (400Hz, DMSO-d6) δ 8.71 (d, J=4.4 Hz, 1H), 8.39 (d, J=8.0 Hz, 1H), 7.66 (dd, J= 7.6 Hz, 4.8 Hz, 1H), 5.20 (s, 2H), 3.39 (s, 3H), 2.43 (s, 3H).

[0087]

[Example of getting 7-6]

N-(4-Chloro-5-methylisoxazol-3-yl)-N-(methoxymethyl)-2-((triisopropylsilyl)ethynyl)pyridine-3-sulfonamide

Under argon atmosphere, bis(triphenylphosphine)palladium(II) dichloride (239 mg, 0.34 mmol) and copper(I) iodide (20 mg, 0.10 mmol) were added to a mixture of 2-chloro-N-(4-chloro-5-methylisoxazol-3-yl)-N-(methoxymethyl)pyridine-3-sulfonamide (1.20 g, 3.41 mmol), triethylamine (1.44 mL, 10.2 mmol), triisopropylsilylacetylene (1.24 g, 6.83 mmol) and tetrahydrofuran (15 mL), and the reaction mixture was stirred at 55°C for 6 h. The reaction mixture was returned to room temperature and ice-cooled water was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.70 g).

1H NMR (400Hz, DMSO-d6) δ 8.83 (d, J=3.6 Hz, 1H), 8.28 (t, J=7.2 Hz, 1H), 7.62-7.66 ( m, 1H), 5.19 (s, 2H), 3.33 (d, J=6.0 Hz, 3H), 2.43 (s, 3H), 1.10 (d, J=3.6 Hz, 21H).

[0088]

[Example of getting 7-7]

N-(4-Chloro-5-methylisoxazol-3-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide

To a mixture of N-(4-chloro-5-methylisoxazol-3-yl)-N-(methoxymethyl)-2-((triisopropylsilyl)ethynyl)pyridine-3-sulfonamide (0.70 g, 1.4 mmol) and tetrahydrofuran (10 mL) was gradually added tetrabutylammonium fluoride (1 mol/L tetrahydrofuran solution, 2.81 mL, 2.81 mmol) at 0°C, and the reaction mixture was stirred at room temperature for 2 h. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.38 g).

ESI-MS: m/z 342.15 [M+1]+

[0089]

[Example of getting 7-8]

N-(4-Chloro-5-methylisoxazol-3-yl)-N-(methoxymethyl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide

Under argon atmosphere, 6-iodo-1,1,5-trimethyl-1,3-dihydroisobenzofuran (0.32 g, 1.11 mmol), copper(I) iodide (6.35 mg, 0.03 mmol), triethylamine (0.46 mL, 3.34 mmol) and tetrakis(triphenylphosphine)palladium(0) (128 mg, 0.11 mmol) were added to a mixture of N-(4-chloro-5-methylisoxazol-3-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide (0.38 g, 1.11 mmol) and toluene (5 mL), and the reaction mixture was stirred at 65 °C for 6 h. The reaction mixture was returned to room temperature and water was added to it, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.25 g).

ESI-MS: m/z 502.10 [M+1]+

[0090]

[Example 7]

N-(4-Chloro-5-methylisoxazol-3-yl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide

To a mixture of N-(4-chloro-5-methylisoxazol-3-yl)-N-(methoxymethyl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide (0.25 g, 0.49 mmol) and methanol (3 ml) was added 6 mol/L hydrochloric acid (3 ml) at 0°C, and the reaction mixture was stirred at 60°C for 16 h. The solvent in the reaction mixture was distilled off under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to the residue, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (35 mg).

ESI-MS: m/z 458.08 [M+1]+

1H-NMR (400Hz, DMSO-d6) δ 8.52 (d, J=3.6 Hz, 1H), 8.19 (d, J=8.0, 1H), 7.46 (s, 1H) , 7.39 (dd, J=8.0, 4.8 Hz, 1H), 7.18 (s, 1H), 4.94 (s, 2H), 2.54 (s, 3H), 2.11 (s, 3H), 1.42 (s, 6H).

[0091]

[Example of obtaining 8-1]

2-Chloro-N-(3,4-dimethylisoxazol-5-yl)pyridine-3-sulfonamide

To a mixture of 3,4-dimethylisoxazol-5-amine (1.0 g, 8.9 mmol) and pyridine (10 ml) were gradually added 4-dimethylaminopyridine (108 mg, 0.89 mmol) and 2-chloropyridine-3-sulfonyl chloride (2.8 g, 13.2 mmol) at room temperature, and the reaction mixture was stirred at 50°C for 16 h. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with 2N hydrochloric acid and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (2.0 g).

ESI-MS: m/z 287.9 [M+1]+

[0092]

[Example of obtaining 8-2]

2-Chloro-N-(3,4-dimethylisoxazol-5-yl)-N-(methoxymethyl)pyridine-3-sulfonamide

To a mixture of 2-chloro-N-(3,4-dimethylisoxazol-5-yl)pyridine-3-sulfonamide (2.0 g, 7.0 mmol) and N,N-dimethylformamide (10 ml) was gradually added 60% sodium hydride (334 mg, 13.9 mmol) at 0°C, and the mixture was stirred at the same temperature as above for 30 min. To the reaction mixture was gradually added chloromethyl methyl ether (0.83 g, 10.5 mmol), and the mixture was stirred at room temperature for 2 h. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by column chromatography on silica gel (hexane/ethyl acetate) to give the title compound (2.0 g).

ESI-MS: m/z 332.18 [M+1]+

[0093]

[Example of getting 8-3]

N-(3,4-Dimethylisoxazol-5-yl)-N-(methoxymethyl)-2-((triisopropylsilyl)ethynyl)pyridine-3-sulfonamide

Under argon atmosphere, bis(triphenylphosphine)palladium(II) dichloride (150 mg, 0.604 mmol) and copper(I) iodide (114 mg, 0.604 mmol) were added to a mixture of 2-chloro-N-(3,4-dimethylisoxazol-5-yl)-N-(methoxymethyl)pyridine-3-sulfonamide (2.0 g, 6.0 mmol), triethylamine (1.8 g, 18 mmol), triisopropylsilylacetylene (1.65 g, 9.06 mmol) and N,N-dimethylformamide (10 mL), and the reaction mixture was stirred at 100 °C for 16 h. The reaction mixture was returned to room temperature and ice-cooled water was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (1.5 g).

ESI-MS: m/z 478.1 [M+1]+

[0094]

[Example of getting 8-4]

N-(3,4-Dimethylisoxazol-5-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide

To a mixture of N-(3,4-dimethylisoxazol-5-yl)-N-(methoxymethyl)-2-((triisopropylsilyl)ethynyl)pyridine-3-sulfonamide (1.0 g, 2.1 mmol) and tetrahydrofuran (10 mL) was gradually added tetrabutylammonium fluoride (1 mol/L tetrahydrofuran solution, 3.1 mL, 3.1 mmol) at 0°C, and the reaction mixture was stirred at room temperature for 1 h. Water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (350 mg).

ESI-MS: m/z 322.0 [M+1]+

[0095]

[Example of getting 8-5]

N-(3,4-Dimethylisoxazol-5-yl)-N-(methoxymethyl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide

Under argon atmosphere, copper(I) iodide (17 mg, 0.09 mmol), triethylamine (282 mg, 2.79 mmol) and tetrakis(triphenylphosphine)palladium(0) (103 mg, 0.09 mmol) were added to a mixture of N-(3,4-dimethylisoxazol-5-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide (300 mg, 0.93 mmol) and N,N-dimethylformamide (3 ml), 6-iodo-1,1,5-trimethyl-1,3-dihydroisobenzofuran (296 mg, 1.02 mmol), and the reaction mixture was stirred at room temperature for 1 h. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (250 mg).

ESI-MS: m/z 482.39 [M+1]+

[0096]

[Example 8]

N-(3,4-Dimethylisoxazol-5-yl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide

To a mixture of N-(3,4-dimethylisoxazol-5-yl)-N-(methoxymethyl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide (200 mg, 0.415 mmol) and methanol (4 ml) was added 50% sulfuric acid (4 ml) at 0°C, and the reaction mixture was stirred at 50°C for 1 h. The solvent in the reaction mixture was distilled off under reduced pressure, and saturated aqueous sodium bicarbonate solution was added to the residue, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high performance liquid chromatography (Luna C18 (250×50 mm) 10 μm, 0.1% aqueous formic acid/acetonitrile) to afford the title compound (60 mg).

ESI-MS: m/z 438.31 [M+1]+

1H NMR (400 MHz, DMSO-d6): δ 11.25 (s, 1H), 8.79 (s, 1H), 8.30 (d, J= 7.2 Hz, 1H), 7.609 (s, 1H), 7.390 (s, 1H), 7.224 (s, 1H), 4.95 (s, 2H), 2.46 (s, 3H), 1.82 (s, 3H), 1.66 (s, 3H), 1.43 (s, 6H).

[0097]

[Example of receiving 9-1]

2-Chloro-N-(4-chloro-3-methylisoxazol-5-yl)pyridine-3-sulfonamide

To a mixture of 4-chloro-3-methylisoxazol-5-amine (1.5 g, 11.4 mmol) and tetrahydrofuran (15 ml) were gradually added 60% sodium hydride (909 mg, 22.7 mmol) and 2-chloropyridine-3-sulfonyl chloride (2.89 g, 13.6 mmol) at 0°C, and the mixture was stirred at room temperature for 1 h. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (2.5 g).

ESI-MS: m/z 307.9 [M+1]+

[0098]

[Example of obtaining 9-2]

2-Chloro-N-(4-chloro-3-methylisoxazol-5-yl)-N-(methoxymethyl)pyridine-3-sulfonamide

To a mixture of 2-chloro-N-(4-chloro-3-methylisoxazol-5-yl)pyridine-3-sulfonamide (1.8 g, 5.8 mmol) and N,N-dimethylformamide (20 ml) was gradually added 60% sodium hydride (467 mg, 11.7 mmol) at 0°C, and the mixture was stirred at the same temperature as above for 30 min. To the reaction mixture was gradually added chloromethyl methyl ether (0.54 ml, 8.8 mmol), and the mixture was stirred at room temperature for 2 h. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by column chromatography on silica gel (hexane/ethyl acetate) to give the title compound (1.3 g).

ESI-MS: m/z 352.27 [M+1]+

[0099]

[Example of getting 9-3]

N-(4-Chloro-3-methylisoxazol-5-yl)-N-(methoxymethyl)-2-((triisopropylsilyl)ethynyl)pyridine-3-sulfonamide

Under argon atmosphere, bis(triphenylphosphine)palladium(II) dichloride (134 mg, 0.184 mmol) and copper(I) iodide (140 mg, 0.738 mmol) were added to a mixture of 2-chloro-N-(4-chloro-3-methylisoxazol-5-yl)-N-(methoxymethyl)pyridine-3-sulfonamide (1.3 g, 3.7 mmol), triethylamine (1.46 mL, 11.1 mmol), triisopropylsilylacetylene (0.8 g, 4.4 mmol) and N,N-dimethylformamide (6.5 mL), and the reaction mixture was stirred at 100 °C for 4 h. The reaction mixture was returned to room temperature and filtered through celite and washed with ethyl acetate. The filtrate was washed with water and saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (600 mg).

ESI-MS: m/z 498.51 [M+1]+

[0100]

[Example of getting 9-4]

N-(4-Chloro-3-methylisoxazol-5-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide

To a mixture of N-(4-chloro-3-methylisoxazol-5-yl)-N-(methoxymethyl)-2-((triisopropylsilyl)ethynyl)pyridine-3-sulfonamide (600 mg, 1.20 mmol) and tetrahydrofuran (10 mL) was gradually added tetrabutylammonium fluoride (1 mol/L tetrahydrofuran solution, 1.2 mL, 1.2 mmol) at 0°C, and the reaction mixture was stirred at room temperature for 2 h. Water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (220 mg).

ESI-MS: m/z 342.0 [M+1]+

[0101]

[Example of getting 9-5]

N-(4-Chloro-3-methylisoxazol-5-yl)-N-(methoxymethyl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide

Under argon atmosphere, 6-iodo-1,1,5-trimethyl-1,3-dihydroisobenzofuran (232 mg, 0.81 mmol), copper(I) iodide (27 mg, 0.15 mmol), triethylamine (0.48 mL, 3.66 mmol) and tetrakis(triphenylphosphine)palladium(0) (84 mg, 0.073 mmol) were added to a mixture of N-(4-chloro-3-methylisoxazol-5-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide (250 mg, 0.73 mmol) and N,N-dimethylformamide (5 mL), and the reaction mixture was stirred at room temperature for 30 min. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (200 mg).

ESI-MS: m/z 502.56 [M+1]+

[0102]

[Example 9]

N-(4-Chloro-3-methylisoxazol-5-yl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide

To a mixture of N-(4-chloro-3-methylisoxazol-5-yl)-N-(methoxymethyl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide (200 mg, 0.39 mmol) and methanol (2 ml) was added 50% sulfuric acid (2 ml) at 0°C, and the reaction mixture was stirred at room temperature for 16 h. The solvent in the reaction mixture was distilled off under reduced pressure, and saturated aqueous sodium bicarbonate solution was added to the residue, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was recrystallized from diethyl ether to give the title compound (35 mg).

ESI-MS: m/z 458.36 [M+1]+

1H NMR (400 MHz, DMSO-d6): δ 8.69 (d, J=3.6 Hz, 1H), 8.30 (dd, J=8.0 Hz, 1.2 Hz, 1H), 7 .55-7.52 (m, 1H), 7.49 (s, 1H), 7.07 (s, 1H), 4.94 (s, 2H), 2.49 (s, 3H), 1.89 (s, 3H), 1.42 (s, 6H).

[0103]

[Example of getting 10-1]

2-Chloro-N-(3-methoxy-5-methylpyrazin-2-yl)pyridine-3-sulfonamide

To a mixture of 3-methoxy-5-methylpyrazin-2-amine (0.2 g, 1.4 mmol) and pyridine (5 ml) were gradually added 4-dimethylaminopyridine (17.5 mg, 0.143 mmol) and 2-chloropyridine-3-sulfonyl chloride (366 mg, 1.73 mmol) at room temperature, and the reaction mixture was stirred at 70°C for 16 h. The reaction mixture was returned to room temperature, and ice-cooled water was added thereto, which was then extracted with ethyl acetate. The organic layer was washed with 6 N hydrochloric acid and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.16 g).

ESI-MS: m/z 315.0 [M+1]+

[0104]

[Example of getting 10-2]

2-Chloro-N-(3-methoxy-5-methylpyrazin-2-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)pyridine-3-sulfonamide

To a mixture of 2-chloro-N-(3-methoxy-5-methylpyrazin-2-yl)pyridine-3-sulfonamide (50 mg, 0.16 mmol) and N,N-dimethylformamide (2 ml) was gradually added potassium carbonate (44 mg, 0.32 mmol) at 0°C, and the mixture was stirred at the same temperature as above for 30 min. To the reaction mixture was gradually added 2-(trimethylsilyl)ethoxymethyl chloride (0.1 ml, 0.24 mmol), and the mixture was stirred at room temperature for 30 min. Ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (40 mg).

ESI-MS: m/z 445.0 [M+1]+

[0105]

[Example of getting 10-3]

N-(3-Methoxy-5-methylpyrazin-2-yl)-2-((triisopropylsilyl)ethynyl)-N-((2-(trimethylsilyl)ethoxy)methyl)pyridine-3-sulfonamide

Under argon atmosphere, bis(triphenylphosphine)palladium(II) dichloride (63 mg, 0.089 mmol) and copper(I) iodide (34.2 mg, 0.179 mmol) were added to a mixture of 2-chloro-N-(3-methoxy-5-methylpyrazin-2-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)pyridine-3-sulfonamide (0.4 g, 0.9 mmol), triethylamine (0.4 mL, 2.7 mmol), triisopropylsilylacetylene (0.3 mL, 1.3 mmol) and N,N-dimethylformamide (5 mL), and the reaction mixture was stirred at 100°C for 5 h. The reaction mixture was returned to room temperature and filtered through celite and washed with ethyl acetate. The filtrate was washed with water and saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (160 mg).

ESI-MS: m/z 591.55 [M+1]+

[0106]

[Example of getting 10-4]

2-Ethynyl-N-(3-methoxy-5-methylpyrazin-2-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)pyridine-3-sulfonamide

To a mixture of N-(3-methoxy-5-methylpyrazin-2-yl)-2-((triisopropylsilyl)ethynyl)-N-((2-(trimethylsilyl)ethoxy)methyl)pyridine-3-sulfonamide (0.16 g, 0.27 mmol) and tetrahydrofuran (5 mL) was gradually added tetrabutylammonium fluoride (1 mol/L tetrahydrofuran solution, 0.27 mL, 0.27 mmol) at 0°C, and the reaction mixture was stirred at room temperature for 2 h. Water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (80 mg).

ESI-MS: m/z 435.2 [M+1]+

[0107]

[Example of getting 10-5]

N-(3-Methoxy-5-methylpyrazin-2-yl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)-N-((2-(trimethylsilyl)ethoxy)methyl)pyridine-3-sulfonamide

Under argon atmosphere, 6-iodo-1,1,5-trimethyl-1,3-dihydroisobenzofuran (146 mg, 0.506 mmol), copper(I) iodide (9 mg, 0.046 mmol), triethylamine (0.2 mL, 1.38 mmol) and tetrakis(triphenylphosphine)palladium(0) (53 mg, 0.046 mmol) were added to a mixture of 2-ethynyl-N-(3-methoxy-5-methylpyrazin-2-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)pyridine-3-sulfonamide (200 mg, 0.460 mmol) and N,N-dimethylformamide (5 mL), and the reaction mixture was stirred at room temperature for 1 h. Ice-cooled water was added to the reaction mixture, which was then extracted ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (170 mg).

ESI-MS: m/z 595.4 [M+1]+

[0108]

[Example 10]

N-(3-Methoxy-5-methylpyrazin-2-yl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)pyridine-3-sulfonamide

To a mixture of N-(3-methoxy-5-methylpyrazin-2-yl)-2-((3,3,6-trimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)-N-((2-(trimethylsilyl)ethoxy)methyl)pyridine-3-sulfonamide (200 mg, 0.336 mmol) and methanol (5 ml) was added 50% sulfuric acid (3 ml) at 0°C, and the reaction mixture was stirred at 60°C for 1 h. The solvent in the reaction mixture was distilled off under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to the residue, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high performance liquid chromatography (Luna C18 (250×50 mm) 10 μm, 0.1% aqueous formic acid/acetonitrile) to afford the title compound (35 mg).

ESI-MS: m/z 465.35 [M+1]+

1H NMR (400 MHz, DMSO-d6): δ 10.98 (s, 1H), 8.79 (d, J=4.0 Hz, 1H), 8.36 (dd, J=1.6, 8 ,0 Hz, 1H), 7.62 (dd, J=4,8,8,4 Hz, 1H) 7.51 (s, 1H), 7.22 (s,1H), 4.95 (s, 2H), 3.80 (s, 3H), 2.44 (s, 3H), 2.22 (s, 3H), 1.44 (s , 6H).

[0109]

[Example of obtaining 11-1]

(E)-N'-(2-Hydroxy-4-methylbenzylidene)acetohydrazide

To a mixture of 2-hydroxy-4-methylbenzaldehyde (3.0 g, 22 mmol) and ethanol (40 mL) was added acetohydrazide (1.64 g, 22.1 mmol) at room temperature, and the reaction mixture was stirred at 90°C for 16 h. The solvent from the reaction mixture was distilled off under reduced pressure, and the residue was washed with hexane to give the title compound (3.0 g) as a crude compound. This compound was used in the next reaction without further purification.

[0110]

[Example of obtaining 11-2]

2-Acetyl-4-methylbenzaldehyde

To a mixture of (E)-N'-(2-hydroxy-4-methylbenzylidene)acetohydrazide (4.6 g, 24 mmol) and tetrahydrofuran (50 mL) was added lead(IV) acetate (11.7 g, 26.3 mmol) at 0°C, and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was filtered through celite and washed with ethyl acetate. The filtrate was washed with saturated aqueous sodium bicarbonate and saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (2.0 g).

1H NMR (400 MHz, CDCl3): δ 10.19 (d, J=4 Hz, 1H), 7.84-7.81 (m, 1H), 7.46 (t, J=6.8 Hz, 2H), 2.65 (d, J=4 Hz, 3H), 2.50 (d, J=3.6 Hz, 3H)

[0111]

[Example of getting 11-3]

1-(2-(Hydroxymethyl)-5-methylphenyl)ethan-1-ol

To a mixture of 2-acetyl-4-methylbenzaldehyde (1.4 g, 8.0 mmol), tetrahydrofuran (3.8 mL) and ethanol (11.2 mL) was added sodium borohydride (1.5 g, 40 mmol) at 0°C, and the reaction mixture was stirred at 70°C for 2 h. The reaction temperature was returned to room temperature, and ice-cooled water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium bicarbonate and saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.85 g).

1H NMR (400 MHz, CDCl3): δ 7.27 (s, 1H), 7.20 (d, J=7.6 Hz, 1H), 6.83(d, J=7.2 Hz, 1H) , 5.01-4.91 (m, 3H), 4.53-4.46 (m, 2H), 2.27 (s, 3H), 1.28 (d, J=6.4 Hz, 3H).

[0112]

[Example of getting 11-4]

1,6-Dimethyl-1,3-dihydroisobenzofuran

To a mixture of 1-(2-(hydroxymethyl)-5-methylphenyl)ethan-1-ol (0.90 g, 5.4 mmol) and tetrahydrofuran (10 ml) was gradually added 60% sodium hydride (0.52 g, 13.6 mmol) at 0°C, and the mixture was stirred at the same temperature as above for 30 min. To the reaction mixture was added trimethyl phosphate (1.5 ml, 13.6 mmol) at room temperature, and the mixture was stirred at the same temperature as above for 16 h. Water was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.6 g).

1H NMR (400 MHz, CDCl3): δ 7.12-7.07 (m, 2H), 6.98 (d, J=5.2 Hz, 1H), 5.31-5.28 (m, 1H ), 5.13-5.00 (m, 2H), 2.39 (d, J=5.2 Hz, 3H), 1.52-1.48 (m, 3H).

[0113]

[Example of getting 11-5]

5-Bromo-1,6-dimethyl-1,3-dihydroisobenzofuran

To a mixture of 1,6-dimethyl-1,3-dihydroisobenzofuran (600 mg, 4.05 mmol) and dichloromethane (10 mL) were added N-bromosuccinimide (721 mg, 4.05 mmol) and trifluoromethanesulfonic acid (1.8 g, 12.2 mmol) at 0°C, and the mixture was stirred at room temperature for 2 h. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate) to give the title compound (0.3 g).

1H NMR (400 MHz, CDCl3): δ 7.51 (d, J=5.2Hz, 1H), 7.26 (d, J=5.2 Hz, 1H), 5.17-5.12 (m , 1H), 4.96 (d, J=12.8 Hz, 1H), 4.88-4.84 (m, 1H), 2.35 (d, J=5.2 Hz, 3H), 1.37 (t, J=6 Hz, 3H).

[0114]

[Example of getting 11-6]

2-((1,6-Dimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)-N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)pyridin-3-sulfonamide

Under argon atmosphere, 5-bromo-1,6-dimethyl-1,3-dihydroisobenzofuran (0.337 g, 1.49 mmol), copper(I) iodide (47 mg, 0.249 mmol), triethylamine (0.36 mL, 2.49 mmol) and tetrakis(triphenylphosphine)palladium(0) (143 mg, 0.124 mmol) were added to a mixture of N-(4,5-dimethylisoxazol-3-yl)-2-ethynyl-N-(methoxymethyl)pyridine-3-sulfonamide (0.40 g, 1.24 mmol) and N,N-dimethylformamide (5 mL), and the reaction mixture was stirred at 80 °C for 4 h. The reaction mixture was returned to room temperature and water was added to it, then extracted ethyl acetate. The organic layer was washed with saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high performance liquid chromatography (Luna C18 (250×50 mm) 10 μm, 0.1% aqueous formic acid/acetonitrile) to give the title compound (25 mg).

ESI-MS: m/z 468.23 [M+1]+

[0115]

[Example 11]

2-((1,6-Dimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)-N-(4,5-dimethylisoxazol-3-yl)pyridin-3-sulfonamide

To a mixture of 2-((1,6-dimethyl-1,3-dihydroisobenzofuran-5-yl)ethynyl)-N-(4,5-dimethylisoxazol-3-yl)-N-(methoxymethyl)pyridine-3-sulfonamide (30 mg, 0.064 mmol) and methanol (5 ml) was added 6 mol/L hydrochloric acid (0.5 ml) at 0°C, and the reaction mixture was stirred at 50°C for 4 h. The reaction mixture was returned to room temperature, and ice-cooled water was added thereto, followed by extraction with ethyl acetate. The organic layer was washed with saturated aqueous sodium bicarbonate and saturated brine and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by high performance liquid chromatography (Luna C18 (250×50 mm) 10 μm, 0.1% aqueous formic acid/acetonitrile) to afford the title compound (5 mg).

ESI-MS: m/z 424.0 [M+1]+

1H NMR (400 MHz, DMSO-d6): δ 11.02 (s, 1H), 8.81 (d, J=5.2 Hz, 1H), 8.34 (t, J=8 Hz, 1H) , 7.63 (t, J=4.4 Hz, 1H), 7.48 (d, J=5.6 Hz, 1H), 7.26 (d, J=5.6 Hz, 1H), 5.22(d, J=5.6 Hz, 1H), 5.05-5.00 (m,1H), 4.92 (t , J=6.4 Hz, 1H), 2.48 (d, J=8.8 Hz, 3H), 2.09 (d, J=6.4 Hz, 3H), 1.81 (d, J=6.4 Hz, 3H), 1.41 (t, J=6 Hz, 3H).

[0116]

[Comparative example 1]

In Comparative Example 1, sitaxentan, described by the following formula, was used.

[0117]

[Comparative example 2]

In Comparative Example 2, a compound described by the following formula was used. This compound is the compound described in Example 1 in International Publication No. WO 2013/115162.

[0118]

[Study Example 1-1: Inhibitory effect on endothelin receptor A]

The inhibitory effect on endothelin receptor A was studied using the following method.

CHO-K1-mt aequorin cells engineered to express human endothelin receptor A (Accession Number NP_001948.1) were grown in antibiotic-free medium for 18 h, then treated with PBS-EDTA (5 mM EDTA), centrifuged (2 min, 405×g, room temperature), and then suspended in assay buffer (DMEM/HAM's F12 with HEPES+0.1% protease-free BSA).

[0119]

Cells at 1× ¹⁰⁶ cells/mL were incubated at room temperature for 4 h in the presence of coelenterazine h (Molecular Probes, Inc.) (final concentration: 5 μM). Endothelin-based agonist response was used to determine the endothelin concentration corresponding to the EC80.

[0120]

Then, 50 μl (10,000 cells/well) of the cell suspension was treated with coelenterazine h and 50 μl of assay buffer containing the test substance (final concentration: 0.5% in DMSO) was added to each well of a 96-well plate. After 15 min, endothelin was added so that the final concentration was the concentration corresponding to the EC80. The activity toward the receptor was measured by the emitted light radiation in an FDSS6000 (Hamamatsu Photonics K.K.). IC50 values were calculated using XLfit software (ID Business Solutions Ltd. (IDBS)).

[0121]

A sample having an IC50 value of 1.0 nM or less or exhibiting an inhibitory effect of 50% or more at 1.0 nM was designated as "A"; a sample having an IC50 value of more than 10 nM or exhibiting an inhibitory effect of less than 50% at 10 nM was designated as "C"; a sample with characteristics intermediate between those of "A" and "C" was designated as "B". The results are shown in Table 1-1.

[Table 1-1]

Table 1-1: Inhibitory effect on the ETA receptor

Example 1 A Example 2 A Example 3 A Example 4 A Example 5 B Example 6 B Example 7 A Example 8 A Example 9 A Example 10 A Example 11 B Comparative example 1 B Comparative example 2 A

[0122]

[Study Example 1-2: Inhibitory effect on endothelin receptor B]

The inhibitory effect on endothelin receptor B was studied using the following method.

CHO-K1-mt aequorin cells engineered to express human endothelin receptor B (Accession Number NP_000106.1) were grown in antibiotic-free medium for 18 h, then treated with PBS-EDTA (5 mM EDTA), centrifuged (2 min, 405×g, room temperature), and then suspended in assay buffer (DMEM/HAM's F12 with HEPES+0.1% protease-free BSA).

[0123]

Cells at 1× ¹⁰⁶ cells/mL were incubated at room temperature for 4 h in the presence of coelenterazine h (Molecular Probes, Inc.) (final concentration: 5 μM). Endothelin-based agonist response was used to determine the endothelin concentration corresponding to the EC80.

[0124]

Then, 50 μl (10,000 cells/well) of the cell suspension was treated with coelenterazine h and 50 μl of assay buffer containing the test substance (final concentration: 0.5% in DMSO) was added to each well of a 96-well plate. After 15 min, endothelin was added so that the final concentration was the concentration corresponding to the EC80. The activity toward the receptor was measured by the emitted light radiation in an FDSS6000 (Hamamatsu Photonics K.K.).

[0125]

A sample having an IC50 value of 1,000 nM or less or exhibiting an inhibitory effect of 50% or more at 1,000 nM was designated as “A”; a sample having an IC50 value of more than 5,000 nM or exhibiting an inhibitory effect of less than 50% at 5,000 nM was designated as “C”; a sample with characteristics intermediate between those of “A” and “C” was designated as “B”. The results are shown in Table 1-2.

[Table 1-2]

Table 1-2: Inhibitory effect on ETB receptor

Example 1 C Example 2 C Example 3 C Example 4 C Example 5 C Example 6 C Example 7 57% inhibition at 5000 nM Example 8 C Example 9 C Example 10 C Example 11 C Comparative example 1 C Comparative example 2 C

[0126]

As in the case of ETA selectivity, a sample that met any of the following criteria was designated as "++".

1. The ratio of ETB IC50 value/ETA IC50 value is greater than 5000

2. Having an inhibitory effect on ETA of 50% or more at 1.0 nM and an inhibitory effect on ETB of less than 50% at 5000 nM

The sample that had an ETB IC50/ETA IC50 value ratio of more than 1000 and 5000 or less was designated as "+".

The results are shown in Tables 1-3.

[Table 1-3]

Table 1-3: Selectivity to ETA (ETB/ETA)

Example 1 ++ Example 2 ++ Example 3 ++ Example 4 ++ Example 8 ++ Example 9 ++ Example 10 ++ Comparative example 1 ++ Comparative example 2 ++

[0127]

[Study Example 2: CYP2C9 Inhibitory Effect]

The inhibitory effect of CYP2C9 was studied using the following method.

(1) Frozen liver microsomal fraction was thawed on ice and diluted with potassium phosphate buffer (ppb) to a final concentration of 0.5 mg/mL.

(2) The assay substrate (diclofenac: D6899, Sigma-Aldrich Co. LLC) was added to it so that its final concentration was 2.5 μM.

(3) The mixture was slowly stirred and incubated at 37±1°C for 5 min to obtain a reaction mixture solution.

(4) 801 μL of the reaction mixture solution was placed in each tube, and the test compound was serially diluted with 0.9 μL of DMSO added thereto so that its final concentration was 9.1 to 20,000 nM (1/3 dilutions at 8 concentrations). In this context, 0.9 μL of DMSO was added in the case of the negative control group, and 0.9 μL of sulfaphenazole (S0758, Sigma-Aldrich Co. LLC) adjusted to a final concentration of 1 μM with DMSO was added in the case of the positive control group.

(5) After mixing, 270 µl aliquots were transferred to each of two clean tubes and incubated in a shaking water bath at 37±1°C for 5 min.

(6) The reaction was initiated by adding 30 μl of 10 mM NADPH and the tubes were incubated at 37°C±1°C for 10 min.

(7) The tubes were removed from the water bath by shaking and the reaction was stopped by adding 300 μl of reaction stop solution. After mixing, the tubes were centrifuged at 1021×g at 4°C for 20 min.

(8) The amount of substrate for the metabolism study was determined by LC-MS/MS to calculate the degree of inhibition at each concentration of the test compound.

(9) The IC50 values were calculated using GraphPad Prism5 software. A sample having an IC50 value of 1 μM or less was designated as “++”; a sample having an IC50 value in the range of more than 1 μM to 10 μM or less was designated as “+”; and a sample having an IC50 value of more than 10 μM was designated as “-”. The results are shown in Table 2.

[0128]

[Table 2]

Table 2

CYP2C9 inhibitory effect Example 1 - Example 3 - Example 4 - Example 5 - Example 6 ++ Example 7 + Example 8 - Example 9 + Example 10 - Comparative example 1 ++ Comparative example 2 ++

[0129]

[Study Example 3: Metabolic Stability in Human and Mouse Liver Microsomes]

Metabolic stability in human liver microsomes was studied using the following method.

1. Materials

(1) Human liver microsomes: 20 mg/ml

(2) Test compound: 1.1 mM solution in DMSO

(3) Potassium phosphate buffer solution: 66.7 mM (pH 7.4)

(4) NADPH solution: 10 mM in potassium phosphate buffer solution

(5) Stop solution: 0.5% formic acid solution in acetonitrile containing warfarin as an internal standard

[0130]

2. Methodology

971.5 μL of potassium phosphate buffer solution and 27.5 μL of human liver microsomes were placed in a polyethylene tube and suspended. To it was added 1 μL of the test compound and an aliquot of 180 μL of this mixture was transferred to a fresh tube. The mixture was pre-incubated at 37 °C for 5 min. Then, 20 μL of NADPH solution (for incubation time of 30 min) or 20 μL of potassium phosphate buffer solution (for incubation time of 0 min) were added to it. After incubation, the reaction was stopped by adding 200 μL of stop solution. Then, the tube was centrifuged at 3220 × g for 20 min and the concentration of unchanged form of the test compound in 200 μL of the supernatant was measured by LC-MS/MS. Based on the area of the obtained peak for the unchanged form, the residual content (%) of the unchanged form was calculated using the content at an incubation time of 0 min equal to 100%.

[0131]

Metabolic stability in mouse liver microsomes was studied using the same procedure as above, with the difference that mouse liver microsomes were used instead of human liver microsomes. Residual contents (%) of unchanged form after 30 min are given in Table 3.

[Table 3]

Table 3

Metabolic stability (human) Metabolic stability (mouse) Example 1 59 85 Example 3 47 85 Example 4 49 91 Example 5 63 90 Example 6 56 54 Example 7 44 81 Example 8 73 40 Example 9 28 5 Example 10 66 68

[0132]

[Study Example 4: PAMPA Membrane Permeability Study]

The membrane permeability study was performed using PAMPA. The specific methodology is as follows.

(1) Lecithin at a concentration of 2% (w/v) was added to the dodecane solution and completely dissolved.

(2) 5 µl of lecithin/dodecane solution was added to each well of the donor plate. To do this, the tip of the pipette was positioned so that it did not touch the membrane.

(3) Immediately thereafter, 150 μL of donor solution (5% DMSO-PBS) containing the test compound (10 μM) was added to each well of the donor plate, and 300 μL of aqueous buffer was added to each well of the PTFE acceptor plate.

(4) The donor plate filled with the donor solution containing the compound under study was slowly placed on the acceptor plate and the lower surface of the membrane was brought into contact with the buffer in all wells.

(5) After closing, the apparatus was incubated at room temperature for 16 h.

(6) After incubation, 100 μl of the solution was taken from each upper surface and lower surface of the membrane in the acceptor plate and acetonitrile containing the same amount of internal standard was added to them.

(7) The concentration of the test compound was measured to calculate the permeation rate.

The results are shown in Table 4. A sample having a penetration velocity value of 1× ^10-8 cm/s or less was designated as “C”; a sample having a value in the range of more than 1× ^10-8 cm/s to 10× ^10-8 cm/s or less was designated as “B”; and a sample having a value of more than 10× ^10-8 cm/s was designated as “A”.

[0133]

[Table 4]

Table 4

Membrane permeation rate Example 1 A Example 3 A Example 4 C Example 5 B Example 6 A Example 7 A Example 8 B Example 9 A Example 10 A

[0134]

[Study Example 5: Study of BigET-1-induced Blood Pressure]

In each male Wistar rat aged 8 to 20 weeks, the trachea was probed with an airway catheter, the carotid artery was catheterized for blood pressure measurement, the femoral vein was catheterized for BigET-1 administration, and the duodenum was catheterized for test substance administration under analgesic and urethane anesthesia. After confirming that blood pressure and heart rate had become stable, diluent (0.5% methylcellulose 400) or 30 mg/kg (5 ml/kg) of test substance made up to volume with diluent were injected into the duodenum. BigET-1 (20 μg/kg) was injected into the duodenum 1 h later and pressure was monitored for 60 min after BigET-1 administration. The study was performed using n=6 rats per group. In the study, systolic blood pressure and diastolic blood pressure were determined every 5 min for 20 minutes, starting 5 min before BigET-1 administration and continuing until 60 min after BigET-1 administration, and AUC0-60 min (mmHg×min) during 60 min after BigET-1 administration was calculated using the change from the value obtained 5 min before BigET-1 administration. Student's t-test was used to compare AUC between the vehicle-treated group and the test substance-treated group.

[Table 5]

Table 5

Change in systolic blood pressure AUC 0-60 min Change in diastolic blood pressure AUC 0-60 min Group of animals injected with a diluent 3057±1273 3191±955 Example 1 83±508 # 278±539 # Example 3 113±565 # 322±435 #

Mean ±SD, N=6, #: p<0.01 compared to vehicle-treated group

Claims (38)

1. A compound described by the following formula (1): Where R ¹ and R ² all independently represent hydrogen or alkyl, R ³ and R ⁴ are all independently hydrogen or alkyl, or R ³ and R ⁴ together form an oxo group, R ⁵ -R ⁷ all independently represent hydrogen, alkyl, haloalkyl, halogen, alkoxy or haloalkoxy, each R ⁸ is independently alkyl, haloalkyl or halogen, n is an integer between 0 and 3, and Ar is described by the following formula (Ar1) or (Ar2): Where X and Y represent nitrogen and oxygen respectively or oxygen and nitrogen respectively, R ⁹ and R ¹⁰ all independently represent hydrogen, alkyl, haloalkyl or halogen, each R ¹¹ is independently alkyl, haloalkyl, halogen, alkoxy or haloalkoxy and m is an integer between 0 and 3, Where: the alkyl represented by each of R ¹ , R ² , R ³ , R ⁴ , R ⁵ -R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ is alkyl having from 1 to 6 carbon atoms, wherein the haloalkyl represented by each of R ⁵ -R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ is a haloalkyl having from 1 to 6 carbon atoms, wherein the alkoxy represented by each of R ⁵ -R ⁷ and R ¹¹ is an alkoxy having from 1 to 6 carbon atoms, where the haloalkoxy represented by each of R ⁵ -R ⁷ and R ¹¹ is a haloalkoxy having from 1 to 6 carbon atoms or a pharmaceutically acceptable salt thereof. 2. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein each of R ¹ and R ² is hydrogen. 3. A compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein each of R ⁵ and R ⁷ is hydrogen and R ⁶ is alkyl having from 1 to 6 carbon atoms. 4. A compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, wherein n is 0. 5. A compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein Ar is described by the formula (Ar1). 6. A compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein each of R ⁹ and R ¹⁰ is alkyl having from 1 to 6 carbon atoms. 7. A compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein X and Y are nitrogen and oxygen, respectively. 8. A compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein X and Y are oxygen and nitrogen, respectively. 9. A compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein Ar is described by the formula (Ar2). 10. A compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 and 9, wherein each R ¹¹ is independently an alkyl having from 1 to 6 carbon atoms or an alkoxy group having from 1 to 6 carbon atoms. 11. A compound or pharmaceutically acceptable salt thereof according to any one of claims 1-4, 9 and 10, wherein m is 2. 12. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is selected from the group consisting of the following compounds: 13. An endothelin A receptor antagonist for the prevention or treatment of a disease associated with the endothelin A receptor, comprising an effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-12. 14. An endothelin receptor A antagonist according to claim 13, wherein the endothelin receptor A antagonist selectively inhibits endothelin receptor A as opposed to endothelin receptor B. 15. A pharmaceutical composition for the prevention or treatment of a disease associated with an endothelin A receptor, comprising an effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-12, wherein the disease associated with an endothelin A receptor is selected from the group consisting of the following: pulmonary hypertension, nephropathy, hypertension, hepatitis, cancer, pain, complications associated with an autoimmune disease, heart failure and vasospasm.