WO2020053249A1 - Flavone compounds for the treatment and prophylaxis of hepatitis b virus disease - Google Patents

Flavone compounds for the treatment and prophylaxis of hepatitis b virus disease Download PDF

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Publication number
WO2020053249A1
WO2020053249A1 PCT/EP2019/074173 EP2019074173W WO2020053249A1 WO 2020053249 A1 WO2020053249 A1 WO 2020053249A1 EP 2019074173 W EP2019074173 W EP 2019074173W WO 2020053249 A1 WO2020053249 A1 WO 2020053249A1
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WO
WIPO (PCT)
Prior art keywords
chloro
ethoxy
chromen
oxo
phenoxy
Prior art date
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PCT/EP2019/074173
Other languages
French (fr)
Inventor
Song Feng
Chungen Liang
Yongfu Liu
Hong Shen
Xuefei Tan
Jun Wu
Dongdong Chen
Chao Li
Li Wang
Original Assignee
F. Hoffmann-La Roche Ag
Hoffmann-La Roche Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by F. Hoffmann-La Roche Ag, Hoffmann-La Roche Inc. filed Critical F. Hoffmann-La Roche Ag
Priority to CN201980056010.8A priority Critical patent/CN112601743B/en
Priority to EP19769742.8A priority patent/EP3849966B1/en
Priority to JP2021512942A priority patent/JP2022500373A/en
Priority to US17/275,502 priority patent/US12012390B2/en
Publication of WO2020053249A1 publication Critical patent/WO2020053249A1/en

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
    • C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
    • C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
    • C07D311/30—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only not hydrogenated in the hetero ring, e.g. flavones
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D335/00—Heterocyclic compounds containing six-membered rings having one sulfur atom as the only ring hetero atom
    • C07D335/04—Heterocyclic compounds containing six-membered rings having one sulfur atom as the only ring hetero atom condensed with carbocyclic rings or ring systems
    • C07D335/06—Benzothiopyrans; Hydrogenated benzothiopyrans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond

Definitions

  • the present invention relates to organic compounds useful for therapy and/or prophylaxis of HBV infection in a mammal, and in particular to cccDNA (covalently closed circular DNA) inhibitors useful for treating HBV infection.
  • cccDNA covalently closed circular DNA
  • the present invention relates to novel flavone derivatives having pharmaceutical activity, their manufacture, pharmaceutical compositions containing them and their potential use as medicaments.
  • the present invention relates to compounds of formula (I)
  • R 1 to R 3 , Gi, G 2 , Ai to A 7 and W are as described below, or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
  • Hepatitis B virus (HBV) infection is one of the most prevalent viral infections and is a leading cause of chronic hepatitis. It is estimated that worldwide, around 2 billion people have evidence of past or present infection with HBV. Over 250 million individuals are currently chronically infected with HBV and are therefore at high risk to develop liver fibrosis, cirrhosis and hepatocellular carcinoma (HCC). There are data to indicate -800,000 deaths per year are directly linked to HBV infection ( Lozano, R. et al., Lancet (2012), 380 (9859), 2095-2128; Goldstein, S.T. et al., Int J Epidemiol (2005), 34 (6), 1329-1339).
  • FDA-approved treatments for chronic hepatitis B include two type 1 interferons (IFN) which are IFNalfa-2b and pegylated IFN alfa-2a and six nucleos(t)ide analogues (NAs) which are lamivudine (3TC), tenofovir disoproxil fumarate (TDF), adefovir (ADV), telbivudine (LdT), entecavir (ETV), and vemlidy (tenofovir alafenamide (TAF)).
  • IFN interferons
  • TDF tenofovir disoproxil fumarate
  • ADV adefovir
  • LdT telbivudine
  • ETV entecavir
  • TAF vemlidy
  • IFN treatment is finite, but it is known to have severe side effects, and only a small percentage of patients showed a sustained viro logical response, measured as loss of hepatitis B surface antigen (HBsAg).
  • NAs are inhibitors of the HBV reverse transcriptase, profoundly reduce the viral load in vast majority of treated patients, and lead to improvement of liver function and reduced incidence of liver failure and hepatocellular carcinoma.
  • the treatment of NAs is infinite (Ahmed, M. et al., Drug Discov Today (2015), 20 (5), 548-561; Zoulim, F. and Locarnini, S., Gastroenterology (2009), 137 (5), 1593-1608 el59l-l592).
  • HBV chronic infection is caused by persistence of covalently closed circular (ccc)DNA, which exists as an episomal form in hepatocyte nuclei.
  • cccDNA serves as the template for viral RNA transcription and subsequent viral DNA generation. Only a few copies of cccDNA per liver cell can establish or re-initiate viral replication. Therefore, a complete cure of chronic hepatitis B will require elimination of cccDNA or permanently silencing of cccDNA.
  • cccDNA is intrinsically very stable and currently available therapeutics could not eliminate cccDNA or permanently silence cccDNA (Nassal, M., Gut (2015), 64 (12), 1972-1984; Gish, R.G.
  • Objects of the present invention are novel compounds of formula (I), their manufacture, medicaments based on a compound in accordance with the invention and their production as well as the use of compounds of formula (I) as cccDNA inhibitors and for the treatment or prophylaxis of HBV infection.
  • the compounds of formula (I) show superior anti-HBV activity.
  • the compounds of formula (I) also show good PK profiles.
  • the present invention relates to a compound of formula (I)
  • W is O or S
  • Ai is CH or CR 4 ;
  • a 2 is CH or CR 4 ;
  • a 3 is CH or CR 4 ;
  • a 4 is N, CH or CR 4 ;
  • a 5 is N, CH or CR 4 ;
  • a 6 is CH
  • a 7 is N or CH
  • R 1 is halogen, Ci- 6 alkyl or C3-7cycloalkyl
  • R 2 is H, OH, halogen or Ci- 6 alkoxy
  • R 3 is carboxy or Ci- 6 alkoxycarbonyl
  • R 4 is halogen, OH, CN, Ci- 6 alkyl, C3-7cyclo alkyl or Ci- 6 alkoxy;
  • Gi is Ci- 6 alkyl, hydroxyCi- 6 alkyl or C3-7cycloalkylCi-6alkyl;
  • G 2 is Ci- 6 alkyl, C3-7cyclo alkyl or phenyl
  • the term“Ci- 6 alkyl” alone or in combination signifies a saturated, linear- or branched chain alkyl group containing 1 to 6, particularly 1 to 4 carbon atoms, for example methyl, ethyl, propyl, isopropyl, l-butyl, 2-butyl, tert- butyl and the like.
  • Particular“Ci- 6 alkyl” groups are methyl, ethyl, isopropyl and tert- butyl. More particularly,“Ci- 6 alkyl” group is methyl.
  • C3-7cycloalkyl denotes to a saturated carbon ring containing from 3 to 7 carbon atoms, particularly from 3 to 6 carbon atoms, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.
  • Particular“C3-7cycloalkyl” group is cyclopropyl, cyclobutyl or cyclopentyl.
  • Ci- 6 alkoxy alone or in combination signifies a group Ci- 6 alkyl-0-, wherein the “Ci- 6 alkyl” is as defined above; for example methoxy, ethoxy, propoxy, fs -propoxy, n-butoxy, Ao-butoxy, 2-butoxy, /e/7-butoxy, pentoxy, hexyloxy and the like.
  • Particular“Ci- 6 alkoxy” groups are methoxy, ethoxy and propoxy. More particularly,“Ci- 6 alkoxy” group is methoxy or ethoxy.
  • halogen means fluorine, chlorine, bromine or iodine.
  • carbonyl alone or in combination refers to the group -C(O)-.
  • enantiomer denotes two stereoisomers of a compound which are non- superimpo sable mirror images of one another.
  • diastereomer denotes a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities.
  • the compounds according to the present invention may exist in the form of their pharmaceutically acceptable salts.
  • pharmaceutically acceptable salt refers to conventional acid-addition salts or base-addition salts that retain the biological effectiveness and properties of the compounds of formula (I) and are formed from suitable non-toxic organic or inorganic acids or organic or inorganic bases.
  • Acid-addition salts include for example those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid, and those derived from organic acids such as -toluenesulfonic acid, salicylic acid, methane sulfonic acid, oxalic acid, succinic acid, citric acid, malic acid, lactic acid, fumaric acid, and the like.
  • Base-addition salts include those derived from ammonium, potassium, sodium and, quaternary ammonium hydroxides, such as for example, tetramethyl ammonium hydroxide.
  • the chemical modification of a pharmaceutical compound into a salt is a technique well known to pharmaceutical chemists in order to obtain improved physical and chemical stability, hygroscopicity, flowability and solubility of compounds. It is for example described in Bastin R.J., et al., Organic Process Research & Development 2000, 4, 427-435. Particular are the sodium salts of the compounds of formula (I).
  • Racemates can be separated according to known methods into the enantiomers.
  • diastereomeric salts which can be separated by crystallization are formed from the racemic mixtures by reaction with an optically active acid such as e.g. D- or L-tartaric acid, mandelic acid, malic acid, lactic acid or camphor sulfonic acid.
  • an optically active acid such as e.g. D- or L-tartaric acid, mandelic acid, malic acid, lactic acid or camphor sulfonic acid.
  • the present invention provides (i) a compound having the general formula (I):
  • W is O or S
  • Ai is CH or CR 4 ;
  • a 2 is CH or CR 4 ;
  • a 3 is CH or CR 4 ;
  • a 4 is N, CH or CR 4 ;
  • a 5 is N, CH or CR 4 ;
  • a 6 is CH
  • a 7 is N or CH
  • R 1 is halogen, Ci- 6 alkyl or C3-7cycloalkyl
  • R 2 is H, OH, halogen, Ci- 6 alkoxy, haloCi- 6 alkoxy or phenylCi- 6 alkoxy;
  • R 3 is carboxy or Ci- 6 alkoxycarbonyl
  • R 4 is halogen, OH, CN, Ci- 6 alkyl, C 3-7 cycloalkyl, Ci- 6 alkoxy, oxopyrrolidinyl, morpholinyl or haloCi-6alkyl;
  • Gi is Ci- 6 alkyl, hydroxyCi- 6 alkyl or C 3-7 cycloalkylCi- 6 alkyl;
  • G 2 is Ci- 6 alkyl, C 3-7 cyclo alkyl or phenyl;
  • a another embodiment of the present invention is a compound of formula (I), wherein W is O or S;
  • Ai is CH or CR 4 ;
  • a 2 is CH or CR 4 ;
  • a 3 is CH or CR 4 ;
  • a 4 is N, CH or CR 4 ;
  • a 5 is N, CH or CR 4 ;
  • a 6 is CH
  • a 7 is N or CH
  • R 1 is halogen, Ci- 6 alkyl or C 3-7 cycloalkyl
  • R 2 is H, OH, halogen or Ci- 6 alkoxy
  • R 3 is carboxy or Ci-6alkoxycarbonyl
  • R 4 is halogen, OH, CN, Ci- 6 alkyl, C 3-7 cyclo alkyl or Ci- 6 alkoxy;
  • Gi is Ci- 6 alkyl, hydroxyCi- 6 alkyl or C 3-7 cycloalkylCi- 6 alkyl;
  • G 2 is Ci- 6 alkyl, C 3-7 cyclo alkyl or phenyl;
  • a further embodiment of the present invention is (ii) a compound of formula (I) according to (i), wherein
  • W is O or S
  • Ai is CH or CR 4 ;
  • a 2 is CH or CR 4 ;
  • a 3 is CH or CR 4 ;
  • a 4 is N, CH or CR 4 ;
  • a 5 is N, CH or CR 4 ;
  • a 6 is CH
  • a 7 is N or CH
  • R 1 is Cl, Br, methyl or cyclopropyl
  • R 2 is H, OH, Cl, Br, methoxy, trifluoromethoxy or benzyloxy
  • R 3 is carboxy, methoxycarbonyl or ethoxycarbonyl
  • R 4 is F, Cl, Br, OH, CN, methyl, isopropyl, cyclopropyl, methoxy, ethoxy, isopropoxy, oxopyrrolidinyl, morpholinyl or trifluoromethyl;
  • Gi is cyclobutylmethyl, ethyl, hydroxypropyl, methylethyl or propyl;
  • G 2 is cyclobutyl, cyclohexyl, cyclopentyl, cyclopropyl, dimethylethyl, ethyl, methyl and phenyl;
  • a further embodiment of the present invention is a compound of formula (I), wherein W is O or S;
  • Ai is CH or CR 4 ;
  • a 2 is CH or CR 4 ;
  • a 3 is CH or CR 4 ;
  • a 4 is N, CH or CR 4 ;
  • a 5 is N, CH or CR 4 ;
  • a 6 is CH
  • a 7 is N or CH
  • R 1 is Cl, Br, methyl or cyclopropyl
  • R 2 is H, OH, Cl or methoxy
  • R 3 is carboxy, methoxycarbonyl or ethoxycarbonyl
  • R 4 is F, Cl, Br, OH, CN, methyl, isopropyl, cyclopropyl, methoxy, ethoxy or isopropoxy; Gi is cyclobutylmethyl, ethyl, hydroxypropyl, methylethyl or propyl;
  • G 2 is cyclobutyl, cyclohexyl, cyclopentyl, cyclopropyl, dimethylethyl, ethyl, methyl and phenyl;
  • a further embodiment of the present invention is (iii) a compound of formula (I) or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein Ai is CH.
  • a further embodiment of the present invention is (iv) a compound of formula (I) or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein A 4 is CH or CR 4 .
  • a further embodiment of the present invention is (v) a compound of formula (I) or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein As is CH or CR 4 .
  • a further embodiment of the present invention is (vi) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein A 7 is CH.
  • a further embodiment of the present invention is (vii) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R 1 is halogen.
  • a further embodiment of the present invention is (viii) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R 2 is H.
  • a further embodiment of the present invention is (ix) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R 3 is carboxy.
  • a further embodiment of the present invention is (x) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R 4 is halogen, Ci- 6 alkyl or Ci- 6 alkoxy.
  • a further embodiment of the present invention is (xi) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein Gi is Ci-6alkyl.
  • a further embodiment of the present invention is (xii) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein G 2 is Ci- 6 alkyl or C 3-7 cycloalkyl.
  • Another further embodiment of the present invention is (xiii) a compound of formula (I), wherein
  • W is O or S
  • Ai is CH
  • a 2 is CH or CR 4 ;
  • a 3 is CH or CR 4 ;
  • a 4 is CH or CR 4 ;
  • a 5 is CH or CR 4 ;
  • a 6 is CH
  • a 7 is CH
  • R 1 is halogen
  • R 2 is H
  • R 3 is carboxy;
  • R 4 is halogen, Ci- 6 alkyl or Ci- 6 alkoxy;
  • Gi Ci- 6 alkyl
  • G 2 is Ci- 6 alkyl or C3-7cyclo alkyl
  • Another further embodiment of the present invention is (xiv) a compound of formula (I), wherein
  • W is O or S
  • Ai is CH
  • a 2 is CH or CR 4 ;
  • a 3 is CH or CR 4 ;
  • a 4 is CH or CR 4 ;
  • a 5 is CH or CR 4 ;
  • a 6 is CH
  • a 7 is CH
  • R 1 is Cl, Br
  • R 2 is H
  • R 3 is carboxy
  • R 4 is F, Cl, Br, methyl, ethoxy or methoxy
  • Gi is ethyl or methylethyl
  • G 2 is cyclobutyl or methyl
  • the compounds of the present invention can be prepared by any conventional means. Suitable processes for synthesizing these compounds as well as their starting materials are provided in the schemes below and in the subsequent examples. All substituents, in particular, R 1 to R 3 , X, Gi, G 2 , A 1 to A 7 are defined as below unless otherwise indicated. Furthermore, and unless explicitly otherwise stated, all reactions, reaction conditions, abbreviations and symbols have the meanings well known to a person of ordinary skill in the art.
  • Q is halogen, OTs, OTf or OMs; W is O or S.
  • the final compound of formula I can also be prepared by starting with condensation of substituted ketone IV with substituted aldehyde XIII in the presence of a base, such as KOH, in a suitable solvent, such as ethanol, affords a,b-unsaturated carbonyl compound of formula XIV. Cyclization of compound of formula XIV in the presence of a suitable Lewis acid, such L, in a suitable solvent, such as DMSO, affords final compound of formula I.
  • W is O or S.
  • the intermediate X can also be prepared according to the Scheme 2. Condensation of substituted ketone IV with substituted aldehyde XVI in the presence of a base, such as KOH, in a suitable solvent, such as ethanol, affords a,b-unsaturated ketone intermediate XVII.
  • a base such as KOH
  • a suitable solvent such as ethanol
  • This invention also relates to a process for the preparation of a compound of formula (I) comprising the following step:
  • R 1 , R 2 , R 3 ,W, Ai to A 7 , Gi and G 2 are defined as above;
  • the base in step (a) can be for example NaH
  • the base in step (b) can be for example K 2 CO 3 ;
  • the Lewis acid in step (c) can be for example I 2 .
  • a compound of formula (I) when manufactured according to the above process is also an object of the invention.
  • the invention also relates to a compound of formula (I) for use as therapeutically active substance.
  • Another embodiment provides pharmaceutical compositions or medicaments containing the compounds of the invention and a therapeutically inert carrier, diluent or excipient, as well as methods of using the compounds of the invention to prepare such compositions and medicaments.
  • compounds of formula (I) may be formulated by mixing at ambient temperature at the appropriate pH, and at the desired degree of purity, with physiologically acceptable carriers, i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form.
  • physiologically acceptable carriers i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form.
  • the pH of the formulation depends mainly on the particular use and the concentration of compound, but preferably ranges anywhere from about 3 to about 8.
  • a compound of formula (I) is formulated in an acetate buffer, at pH 5.
  • the compounds of formula (I) are sterile.
  • the compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation or as an aqueous solution.
  • compositions are formulated, dosed, and administered in a fashion consistent with good medical practice.
  • Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners.
  • The“effective amount” of the compound to be administered will be governed by such considerations, and is the minimum amount necessary to inhibit cccDNA in HBV patients, consequently lead to the reduction of HBsAg and HBeAg (HBV e antigen) in serum. For example, such amount may be below the amount that is toxic to normal cells, or the mammal as a whole.
  • the pharmaceutically effective amount of the compound of the invention administered parenterally per dose will be in the range of about 0.1 to 100 mg/kg, alternatively about 0.1 to 50 mg/kg of patient body weight per day, with the typical initial range of compound used being 0.3 to 15 mg/kg/day.
  • oral unit dosage forms such as tablets and capsules, preferably contain from about 25 to about 1000 mg of the compound of the invention.
  • the compounds of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if desired for local treatment, intralesional administration.
  • Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
  • the compounds of the present invention may be administered in any convenient administrative form, e.g., tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc.
  • Such compositions may contain components conventional in pharmaceutical preparations, e.g., diluents, carriers, pH modifiers, sweeteners, bulking agents, and further active agents.
  • a typical formulation is prepared by mixing a compound of the present invention and a carrier or excipient.
  • Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, e.g., Ansel, Howard C., et al., Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R., et al. Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005.
  • the formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament).
  • buffers stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing
  • An example of a suitable oral dosage form is a tablet containing about 25 to 500 mg of the compound of the invention compounded with about 90 to 30 mg anhydrous lactose, about 5 to 40 mg sodium croscarmellose, about 5 to 30 mg polyvinylpyrrolidone (PVP) K30, and about 1 to 10 mg magnesium stearate.
  • the powdered ingredients are first mixed together and then mixed with a solution of the PVP.
  • the resulting composition can be dried, granulated, mixed with the magnesium stearate and compressed to tablet form using conventional equipment.
  • An example of an aerosol formulation can be prepared by dissolving the compound, for example 5 to 400 mg, of the invention in a suitable buffer solution, e.g. a phosphate buffer, adding a tonicifier, e.g. a salt such sodium chloride, if desired.
  • the solution may be filtered, e.g., using a 0.2 micron filter, to remove impurities and contaminants.
  • An embodiment therefore, includes a pharmaceutical composition comprising a compound of Formula (I), or pharmaceutically acceptable salt or enantiomer or diastereomer thereof.
  • composition comprising a compound of formula (I), or pharmaceutically acceptable salt or enantiomer or diastereomer thereof, together with a pharmaceutically acceptable carrier or excipient.
  • Another embodiment includes a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of formula (I), or pharmaceutically acceptable salt or enantiomer or diastereomer thereof for use in the treatment of HBV infection.
  • the compounds of the invention can inhibit cccDNA and have anti-HBV activity. Accordingly, the compounds of the invention are useful for the treatment or prophylaxis of HBV infection.
  • the invention relates to the use of a compound of formula (I) for the inhibition of cccDNA.
  • the invention also relates to the use of a compound of formula (I) for the inhibition of HBeAg.
  • the invention further relates to the use of a compound of formula (I) for the inhibition of HBsAg.
  • the invention relates to the use of a compound of formula (I) for the inhibition of HBV
  • the invention relates to the use of a compound of formula (I) for the treatment or prophylaxis of HBV infection.
  • the invention relates in particular to the use of a compound of formula (I) for the preparation of a medicament for the treatment or prophylaxis of HBV infection.
  • Another embodiment includes a method for the treatment or prophylaxis of HBV infection, which method comprises administering an effective amount of a compound of Formula (I), or enantiomers, diastereomers, prodrugs or pharmaceutically acceptable salts thereof.
  • Figure 1 the result of Example 1 in cccDNA Southern Blot assay, it indicates that Example 1 dose-dependently reduced cccDNA level in HepDESl9 cells.
  • EC50 the molar concentration of an inhibitor, which produces 50% of the maximum possible response for that inhibitor
  • FBS fetal bovine serum
  • H2O2 hydrogen peroxide
  • PE petroleum ether
  • PPA polyphosphoric acid
  • Acidic condition A: 0.1% formic acid and 1% acetonitrile in H 2 0; B: 0.1% formic acid in acetonitrile;
  • Mass spectra generally only ions which indicate the parent mass are reported, and unless otherwise stated the mass ion quoted is the positive mass ion (M+H) + .
  • Step 2 Preparation of .v-ethyl 3-[2-(4-formylphenoxy)ethoxy]cyclobutanecarboxylate
  • Step 1 Preparation of (E)-l-(3-chloro-2-hydroxy-phenyl)-3-[4-(2- hydroxyethoxy)phenyl]prop-2-en-l-one
  • Step 3 Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Example 1-B The configuration of Example 1-A and Example 1-B were determined by NOESY.
  • Example 1-A Example 1-B
  • Step 1 Preparation of (E)-l-(3-chloro-2-hydroxy-phenyl)-3-(4-methoxyphenyl)prop-2-en- 1-one
  • Step 5 Preparation of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] benzoate
  • Step 3 Preparation of ethyl 2-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy]ethoxy]cyclopropanecarboxylate
  • Example 4-A 40 mg, 20.4%
  • Example 4-B 60 mg, 35%)
  • Example 5-A 2.9 mg, 3.6%) and the other is Example 5-B (l7mg, 35.3%).
  • Compound 6a was prepared in analogy to the procedure described for the preparation of Int-1 by using methyl 3-oxocyclopentanecarboxylate as the starting materials instead of methyl 3-oxocyclobutanecarboxylate in Step 2.
  • Step 2 Preparation of methyl 3-[2-(p-tolylsulfonyloxy)ethoxy]cyclopentanecarboxylate
  • Step 3 Preparation of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclopentanecarboxy late
  • Step 4 Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclopentanecarboxy lie acid
  • Compound 7a was prepared in analogy to the procedure described for the preparation of Int-1 by using methyl 4-oxocyclohexanecarboxylate as the starting materials instead of methyl 3-oxocyclobutanecarboxylate in Step 2.
  • Step 2 Preparation of methyl 4-[2-(p-tolylsulfonyloxy)ethoxy]cyclohexanecarboxylate
  • Step 3 Preparation of methyl 4-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclohexanecarboxy late
  • Step 4 Preparation of 4-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclohexanecarboxy lie acid
  • Example 7-A 720 mg, 51.2%) and the other is Example 7-B (30mg, 2.1%).
  • Step 3 Preparation of 8-chloro-2-[4-[(3-hydroxycyclobutyl)methoxy]phenyl]chromen-4- one
  • Step 4 Preparation of ethyl 2-[3-[[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] methyl] cyclobutoxy ] acetate
  • Step 5 Preparation of 2-[3-[[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] methyl] cyclobutoxy] acetic acid
  • Step 1 Preparation of methyl 2-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] acetate
  • Example 10 was prepared in analogy to the procedure described for the preparation of example 9 by using ethyl 3-chloropropanoate as the starting material instead of methyl 2- chloro acetate in Step 1.
  • Step 2 Preparation of methyl 3-(2-chloro-l-methyl-ethoxy)cyclobutanecarboxylate
  • Step 3 Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-l-methyl- ethoxy ] cyclobutanecarboxylic acid
  • Example 12 was prepared in analogy to the procedure described for the preparation of Example 7 by using methyl 3-(2-chloro-l-methyl-ethoxy)cyclobutanecarboxylate as the starting material instead of methyl 4-[2-(p-tolylsulfonyloxy)ethoxy]cyclohexanecarboxylate in Step 3.
  • Step 2 Preparation of 3-[3-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy]propoxy]cyclobutanecarboxylic acid
  • Example 13 was prepared in analogy to the procedure described for the preparation of Example 1 by using 8-chloro-2-[4-(3-hydroxypropoxy)phenyl]chromen-4-one as the starting material instead of 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one in Step 3.
  • Step 2 Preparation of methyl 3-[3-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-2-hydroxy- propoxy ] benzoate
  • Step 2 Preparation of l-(3-chloro-4-fluoro-2-hydroxy-phenyl)ethanone
  • Step 3 Preparation of (/i)-l-(3-chloro-4-fluoro-2-hydroxy-phenyl)-3-(4- methoxyphenyl)prop-2-en-l-one
  • Step 5 Preparation of methyl 3-[2-[4-(8-chloro-7-fluoro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxylate
  • Step 2 Preparation of 3-[2-[4-[(£ 3-(3-chloro-2,4-dihydroxy-phenyl)-3-oxo-prop-l- enyl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Step 3 Preparation of 3-[2-[4-(8-chloro-7-hydroxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Step 4 Preparation of methyl 3-[2-[4-(8-chloro-7-methoxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy late
  • Step 5 Preparation of 3-[2-[4-(8-chloro-7-methoxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxylic acid
  • Compound 20a was prepared in analogy to the procedure described for the preparation of compound 15b by using 2-chloro-4-fluoro-phenol as the starting materials instead of 2-chloro-3- fluoro -phenol in Step 1.
  • Step 2 Preparation of 3-[2-[4-[(E)-3-(3-chloro-5-fluoro-2-hydroxy-phenyl)-3-oxo-prop-l- enyl] phenoxy ] ethoxy] cyclobutanecarboxylic acid
  • Compound 20b was prepared in analogy to the procedure described for the preparation of compound 19b by using l-(3-chloro-5-fluoro-2-hydroxy-phenyl)ethanone as the starting materials instead of l-(3-chloro-2,4-dihydroxy-phenyl)ethanone in Step 2.
  • Step 3 Preparation of 3-[2-[4-(8-chloro-6-fluoro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Example 20 NMR DMSO-d , 400MHz): d ppm 12.18 (s, 1H), 8.11-8.09 (m, 3H), 7.72-7.70 (m, 1H), 7.19-7.17 (m, 2H), 7.09-7.08 (m, 1H), 4.19-3.98 (m, 2H), 3.97-3.93 (m, 1H), 3.68-3.66 (m, 2H), 2.61-2.57 (m, 1H), 2.46-2.42 (m, 2H), 2.03-1.98 (m, 2H). MS obsd. (ESL) [(M+H) + ]: 433.1.
  • Example 21 Example 21
  • Example 21 was prepared in analogy to the procedure described for the preparation of
  • Example 15 by using 2,3-dichlorophenol as the starting materials instead of 2-chloro-3-fluoro- phenol in Step 1.
  • Step 1 Preparation of l-(5-bromo-3-chloro-2-hydroxy-phenyl)ethanone
  • Example 22 was prepared in analogy to the procedure described for the preparation of Example 15 by using 8-chloro-2-(4-hydroxyphenyl)-4-oxo-chromene-6-carbonitrile as the starting materials instead of 8-chloro-7-fluoro-2-(4-hydroxyphenyl)chromen-4-one in Step 5.
  • Example 23 was prepared in analogy to the procedure described for the preparation of Example 15 by using 8-chloro-2-(4-hydroxyphenyl)-4-oxo-chromene-7-carbonitrile as the starting materials instead of 8-chloro-7-fluoro-2-(4-hydroxyphenyl)chromen-4-one in Step 5.
  • 45 mg of 3-[2-[4-(8-chloro-7-cyano-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid was further purified by supercritical fluid chromatography (SFC) to give two diastereomers with cis- and trans- configuration, one of which is characterized as Example 23-A (31 mg) and the other is Example 23-B (11 mg).
  • SFC supercritical fluid chromatography
  • Step 1 Preparation of l-(3-chloro-2,6-dihydroxy-phenyl)ethanone
  • Step 2 Preparation of l-[3-chloro-6-hydroxy-2-(2- methoxyethoxymethoxy)phenyl]ethanone
  • Step 3 Preparation of ,s-3-[2-[4-[(/i)-3-[3-chloro-6-hydroxy-2-(2- methoxyethoxymethoxy)phenyl]-3-oxo-prop-l-enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid
  • Step 4 Preparation of .s-3-[2-[4-(8-chloro-5-hydroxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Example 25 was prepared in analogy to the procedure described for the preparation of Example 19 by using c/5'-3-[2-[4-(8-chloro-5-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid as the starting materials instead of 3-[2-[4-(8- chloro-7-hydroxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
  • Example 27 was prepared in analogy to the procedure described for the preparation of Example 19 by using c/5'-3-[2-[4-(8-chloro-5-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid and 2-iodopropane as the starting materials instead of 3-[2-[4-(8-chloro-7-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (intermediate 19c) and iodomethane in Step 1.
  • Step 1 Preparation of l-(6-bromo-3-chloro-2-hydroxy-phenyl)ethanone
  • Step 2 Preparation of .s-3-[2-[4-[(/i)-3-(6-bronio-3-chloro-2-hydroxy-phenyl)-3-oxo-prop- 1 -enyl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Example 28 was prepared in analogy to the procedure described for the preparation of Example 19 by using cis-3-[2-[4-[(£ ’ )-3-(6-bromo-3-chloro-2-hydroxy-phenyl)-3-oxo-prop-l- enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid as the starting materials instead of 3-[2-[4- [(£)-3-(3-chloro-2, 4-dihydro xy-phenyl)-3-oxo-prop-l- enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid (intermediate 19b) in Step 3.
  • Step 1 Preparation of l-(3,6-dichloro-2-hydroxy-phenyl)ethanone
  • Step 2 Preparation of C3 ⁇ 4-3-[2-[4-(5,8-dichloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Example 29 was prepared in analogy to the procedure described for the preparation of Example 19 by using l-(3,6-dichloro-2-hydroxy-phenyl)ethanone and r/.v-ethyl 3-(2-(4- formylphenoxy)ethoxy)cyclobutanecarboxylate as the starting materials instead of l-(3-chloro- 2, 4-dihydro xy-phenyl)ethanone (intermediate 19a) and methyl 3 -[2- (4- formylphenoxy)ethoxy]cyclobutanecarboxylate in Step 2.
  • Example 31 was prepared in analogy to the procedure described for the preparation of Example 30 by using 2,4,6-trimethyl- 1,3, 5, 2, 4, 6-trio xatriborinane as the starting materials instead of cyclopropylboronic acid.
  • Example 33 was prepared in analogy to the procedure described for the preparation of Example 19 by using l-(3-bromo-2-hydroxy-phenyl)ethanone and r/.v-ethyl 3-(2-(4- formylphenoxy)ethoxy)cyclobutanecarboxylate as the starting materials instead of l-(3-chloro- 2, 4-dihydro xy-phenyl)ethanone (intermediate 19a) and methyl 3 -[2- (4- formylphenoxy)ethoxy]cyclobutanecarboxylate in Step 2.
  • Example 34 was prepared in analogy to the procedure described for the preparation of Example 30 by using 3-[2-[4-(8-bromo-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid as the starting materials instead of cis-3-[2-[4-(5- bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid.
  • Example 35 was prepared in analogy to the procedure described for the preparation of
  • Example 30 by using 3-[2-[4-(8-bromo-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid and 2,4,6-trimethyl- 1,3, 5, 2, 4, 6-trio xatriborinane as the starting materials instead of cis-3-[2-[4-(5-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid and 2,4,6-trimethyl- 1,3, 5, 2, 4, 6-trio xatriborinane.
  • Step 1 Preparation of l-(3-chloro-6-fluoro-2-hydroxy-phenyl)ethanone
  • Compound 37b was prepared in analogy to the procedure described for the preparation of compound la by using l-(3-chloro-6-fluoro-2-hydroxy-phenyl)ethanone as the starting material instead of l-(3-chloro-2-hydroxy-phenyl)ethanone in Step 1.
  • Step 3 Preparation of 2-[2-[4-(8-chloro-5-fluoro-4-oxo-chromen-2- yl)phenoxy] ethoxy] acetic acid
  • Example 37 was prepared in analogy to the procedure described for the preparation of Example 9 by using 8-chloro-5-fluoro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one as the starting material instead of 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one in Step 1.
  • Step 2 Preparation of l-(3-chloro-2-hydroxy-phenyl)-3-(6-chloro-3-pyridyl)propane-l,3- dione
  • Step 4 Preparation of ,s-3-[2-[[5-(8-chloro-4-oxo-chromen-2-yl)-2-pyridyl]oxy]ethoxy]cyclobutanecarboxylic acid and frans-3-[2-[[5-(8-chloro-4-oxo-chromen-2-yl)-2- pyridy 1] oxy ] ethoxy] cy clobutanecarboxy lie acid
  • Example 38-A l H NMR (DMSO -d 6 , 400MHz): d ppml2.62 (s, 1H), 8.85-8.95 (m, 1H), 8.30-8.38 (m, 1H), 8.00-8.07 (m, 1H), 7.89-7.96 (m, 1H), 7.43-7.51 (m, 1H), 6.97-7.06 (m, 2H), 4.44-4.54 (m, 2H), 3.92-4.04 (m, 1H), 3.63-3.78 (m, 2H), 2.59-2.67 (m, 1H), 2.43-2.53 (m, 2H), 1.98-2.17 (m, 2H). MS obsd. (ESF) [(M+H) + ]: 416.1.
  • Example ppm 11.72-12.61 (m, 1H), 8.87-8.96 (m, 1H), 8.33-8.43 (m, 1H), 7.99 (dd, 7 1.59, 7.83 Hz, 2H), 7.49 (s, 1H), 7.15 (s, 1H), 7.03-7.10 (m, 1H), 4.42-4.53 (m, 2H), 4.09-4.24 (m, 1H), 3.63-3.71 (m, 2H), 2.87-2.98 (m, 1H), 2.34-2.44 (m, 2H), 2.08-2.23 (m, 2H). MS obsd. (ESI + ) [(M+H) + ]: 416.1.
  • Compound 39b was prepared in analogy to the procedure described for the preparation of compound 38c by using 5-chloropyrazine-2-carbonyl chloride as the starting material instead of 6-chloropyridine-3-carbonyl chloride chloride in Step 1.
  • Step 3 Preparation of ci, v-3-[2-[5-(8-chloro-4-oxo-chromen-2-yl)pyrazin-2- yl] oxy ethoxy ]cyclobutanecarboxy lie acid and civ- methyl 3-[2-[5-(8-chloro-4-oxo-chromen- 2-yl)pyrazin-2-yl]oxyethoxy]cyclobutanecarboxylate
  • Compound 41b was prepared in analogy to the procedure described for the preparation of compound 38c by using 6-chloropyridazine-3-carbonyl chloride as the starting material instead of 6-chloropyridine-3-carbonyl chloride chloride in Step 1.
  • Step 3 Preparation of .v- methyl 3-[2-[6-(8-chloro-4-oxo-chromen-2-yl)pyridazin-3- yl] oxy ethoxy ] cyclobutanecarboxy late
  • Step 3 Preparation of .s-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-fluoro- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Step 1 Preparation of 8-chloro-2-(3-chloro-4-methoxy-phenyl)chromen-4-one
  • Compound 43a was prepared in analogy to the procedure described for the preparation of compound 38c by using 3-chloro-4-methoxy-benzoyl chloride as the starting material instead of 6-chloropyridine-3-carbonyl chloride chloride in Step 1.
  • Step 2 Preparation of ' .s-3-[2-[2-chloro-4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Example 43 was prepared in analogy to the procedure described for the preparation of Example 42 by using 8-chloro-2-(3-chloro-4-methoxy-phenyl)chromen-4-one as the starting material instead of 8-chloro-2-(3-fluoro-4-methoxy-phenyl)chromen-4-one in Step 2.
  • Example 43 l H NMR (DMSO -d 6 , 400MHz): d ppm 12.02-12.44 (m, 1H), 8.15-8.21 (m, 1H), 8.05-8.10 (m, 1H), 7.96-8.02 (m, 2H), 7.45-7.54 (m, 1H), 7.34-7.40 (m, 1H), 7.14-7.21 (m, 1H), 4.26-4.31 (m, 2H), 3.92-4.07 (m, 1H), 3.67-3.77 (m, 2H), 2.55-2.63 (m, 1H), 2.39-2.47 (m, 2H), 1.96-2.06 (m, 2H). MS obsd. (ESE) [(M+H) + ]: 449.1.
  • Compound 44a was prepared in analogy to the procedure described for the preparation of compound 3c by using 3-bromo-4-methoxy-benzaldehyde as the starting material instead of 4- methoxybenzaldehyde in Step 1.
  • Step 2 Preparation of methyl 3-[2-[2-bromo-4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy late
  • Step 3 Preparation of ,v-3-[2-[2-bronio-4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Step 1 Preparation of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyclopropyl- phenoxy ] ethoxy] cyclobutanecarboxy late
  • Step 2 Preparation of ,s-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyclopropyl- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Step 1 Preparation of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyano- phenoxy ] ethoxy] cyclobutanecarboxy late
  • Compound 47a was prepared in analogy to the procedure described for the preparation of compound 3c by using 4-methoxy-3-methyl-benzaldehyde as the starting material instead of 4- methoxybenzaldehyde in Step 1.
  • Step 2 Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-methyl- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Example 47 was prepared in analogy to the procedure described for the preparation of
  • Example 15 by using 8-chloro-2-(4-hydroxy-3-methyl-phenyl)chromen-4-one as the starting material instead of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one in Step 5.
  • Step 1 Preparation of 8-chloro-2-(4-hydroxy-3-isopropyl-phenyl)chromen-4-one
  • Compound 48a was prepared in analogy to the procedure described for the preparation of compound 3c by using 3-isopropyl-4-methoxy-benzaldehyde as the starting material instead of 4-methoxybenzaldehyde in Step 1.
  • Step 2 Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-isopropyl- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Example 48 was prepared in analogy to the procedure described for the preparation of Example 15 by using 8-chloro-2-(4-hydroxy-3-isopropyl-phenyl)chromen-4-one as the starting material instead of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one in Step 5.
  • Example 48 NMR (DMSO -d 6 , 400MHz): d ppm 8.04 - 7.91 (m, 4H), 7.53 - 7.43 (m,
  • Compound 49a was prepared in analogy to the procedure described for the preparation of compound 3c by using 2-fluoro-4-methoxy-benzaldehydeas the starting material instead of 4- methoxybenzaldehyde in Step 1.
  • Step 2 Preparation of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-fluoro- phenoxy ] ethoxy] cyclobutanecarboxy late
  • Compound 49b was prepared in analogy to the procedure described for the preparation of compound 15f by using 8-chloro-2-(2-fluoro-4-hydroxy-phenyl)chromen-4-one as the starting material instead of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one in Step 5.
  • Step 2 Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-methoxy- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Step 1 Preparation of (/i)- l-(3-chloro-2-hydroxy-phenyl)-3-(2-ethoxy-4-niethoxy- phenyl)prop-2-en-l-one
  • Compound 51b was prepared in analogy to the procedure described for the preparation of compound 3c by using (£ ’ )-l-(3-chloro-2-hydroxy-phenyl)-3-(2-ethoxy-4-methoxy-phenyl)prop- 2-en-l-one as the starting material instead of (£ ’ )-l-(3-chloro-2-hydroxy-phenyl)-3-(4- methoxyphenyl)prop-2-en-l-one in Step 2.
  • Step 3 Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-ethoxy- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Example 51 was prepared in analogy to the procedure described for the preparation of
  • Example 15 by using 8-chloro-2-(2-ethoxy-4-hydroxy-phenyl)chromen-4-one as the starting material instead of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one in Step 5.
  • Step 1 Preparation of ethyl 3-[2-[4-(3,8-dichloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxylate
  • Step 2 Preparation of 3-[2-[4-(3,8-dichloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
  • Step 2 Preparation of cis-3-[2-[4-(8-chloro-3-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxy lie acid

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Abstract

The present invention provides flavone derivatives having the general formula (I) which are useful for the treatment of Hepatitis B Virus infection (HBV). The compounds act as cccDNA (covalently closed circular DMA) inhibitors.

Description

FLAVONE COMPOUNDS FOR THE TREATMENT AND PROPHYLAXIS OF HEPATITIS B VIRUS DISEASE
The present invention relates to organic compounds useful for therapy and/or prophylaxis of HBV infection in a mammal, and in particular to cccDNA (covalently closed circular DNA) inhibitors useful for treating HBV infection.
FIELD OF THE INVENTION
The present invention relates to novel flavone derivatives having pharmaceutical activity, their manufacture, pharmaceutical compositions containing them and their potential use as medicaments.
The present invention relates to compounds of formula (I)
Figure imgf000002_0001
wherein R1 to R3, Gi, G2, Ai to A7 and W are as described below, or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
Hepatitis B virus (HBV) infection is one of the most prevalent viral infections and is a leading cause of chronic hepatitis. It is estimated that worldwide, around 2 billion people have evidence of past or present infection with HBV. Over 250 million individuals are currently chronically infected with HBV and are therefore at high risk to develop liver fibrosis, cirrhosis and hepatocellular carcinoma (HCC). There are data to indicate -800,000 deaths per year are directly linked to HBV infection ( Lozano, R. et al., Lancet (2012), 380 (9859), 2095-2128; Goldstein, S.T. et al., Int J Epidemiol (2005), 34 (6), 1329-1339).
Many countries in the world administer hepatitis B vaccine starting at birth or in early childhood, which has greatly reduced the incidence and prevalence of hepatitis B in most endemic regions over the past few decades. However the vaccine has no impact on people who were infected before the widely use of the vaccine in developing end-stage liver disease or HCC (Chen, D.S., J Hepatol (2009), 50 (4), 805-816). Vaccination at birth of infants born to HBV positive mothers is usually not sufficient for protecting vertical transmission and combination with hepatitis B immune globulin is needed (Li, X.M. et al., World J Gastroenterol (2003), 9 (7), 1501-1503).
Currently FDA-approved treatments for chronic hepatitis B include two type 1 interferons (IFN) which are IFNalfa-2b and pegylated IFN alfa-2a and six nucleos(t)ide analogues (NAs) which are lamivudine (3TC), tenofovir disoproxil fumarate (TDF), adefovir (ADV), telbivudine (LdT), entecavir (ETV), and vemlidy (tenofovir alafenamide (TAF)). IFN treatment is finite, but it is known to have severe side effects, and only a small percentage of patients showed a sustained viro logical response, measured as loss of hepatitis B surface antigen (HBsAg). NAs are inhibitors of the HBV reverse transcriptase, profoundly reduce the viral load in vast majority of treated patients, and lead to improvement of liver function and reduced incidence of liver failure and hepatocellular carcinoma. However, the treatment of NAs is infinite (Ahmed, M. et al., Drug Discov Today (2015), 20 (5), 548-561; Zoulim, F. and Locarnini, S., Gastroenterology (2009), 137 (5), 1593-1608 el59l-l592).
HBV chronic infection is caused by persistence of covalently closed circular (ccc)DNA, which exists as an episomal form in hepatocyte nuclei. cccDNA serves as the template for viral RNA transcription and subsequent viral DNA generation. Only a few copies of cccDNA per liver cell can establish or re-initiate viral replication. Therefore, a complete cure of chronic hepatitis B will require elimination of cccDNA or permanently silencing of cccDNA. However, cccDNA is intrinsically very stable and currently available therapeutics could not eliminate cccDNA or permanently silence cccDNA (Nassal, M., Gut (2015), 64 (12), 1972-1984; Gish, R.G. et al., Antiviral Res (2015), 121, 47-58; Levrero, M. et al., J Hepatol (2009), 51 (3), 581-592.). The current SoC could not eliminate the cccDNA which are already present in the infected cells. There is an urgent need to discover and develop new anti-HBV reagents to eliminate or permanently silence cccDNA, the source of chronicity (Ahmed, M. et al., Drug Discov Today (2015), 20 (5), 548-561; Nassal, M., Gut (2015), 64 (12), 1972-1984).
SUMMARY OF THE INVENTION
Objects of the present invention are novel compounds of formula (I), their manufacture, medicaments based on a compound in accordance with the invention and their production as well as the use of compounds of formula (I) as cccDNA inhibitors and for the treatment or prophylaxis of HBV infection. The compounds of formula (I) show superior anti-HBV activity. In addition, the compounds of formula (I) also show good PK profiles.
The present invention relates to a compound of formula (I)
Figure imgf000004_0001
wherein,
W is O or S;
Ai is CH or CR4;
A2 is CH or CR4;
A3 is CH or CR4;
A4 is N, CH or CR4;
A5 is N, CH or CR4;
A6 is CH;
A7 is N or CH;
R1 is halogen, Ci-6alkyl or C3-7cycloalkyl;
R2 is H, OH, halogen or Ci-6alkoxy;
R3 is carboxy or Ci-6alkoxycarbonyl;
R4 is halogen, OH, CN, Ci-6alkyl, C3-7cyclo alkyl or Ci-6alkoxy;
Gi is Ci-6alkyl, hydroxyCi-6alkyl or C3-7cycloalkylCi-6alkyl;
G2 is Ci-6alkyl, C3-7cyclo alkyl or phenyl;
or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
DETAILED DESCRIPTION OF THE INVENTION
DEFINITIONS
As used herein, the term“Ci-6alkyl” alone or in combination signifies a saturated, linear- or branched chain alkyl group containing 1 to 6, particularly 1 to 4 carbon atoms, for example methyl, ethyl, propyl, isopropyl, l-butyl, 2-butyl, tert- butyl and the like. Particular“Ci-6alkyl” groups are methyl, ethyl, isopropyl and tert- butyl. More particularly,“Ci-6alkyl” group is methyl.
The term“C3-7cycloalkyl” denotes to a saturated carbon ring containing from 3 to 7 carbon atoms, particularly from 3 to 6 carbon atoms, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. Particular“C3-7cycloalkyl” group is cyclopropyl, cyclobutyl or cyclopentyl.
The term“Ci-6alkoxy” alone or in combination signifies a group Ci-6alkyl-0-, wherein the “Ci-6alkyl” is as defined above; for example methoxy, ethoxy, propoxy, fs -propoxy, n-butoxy, Ao-butoxy, 2-butoxy, /e/7-butoxy, pentoxy, hexyloxy and the like. Particular“Ci-6alkoxy” groups are methoxy, ethoxy and propoxy. More particularly,“Ci-6alkoxy” group is methoxy or ethoxy.
The term“halogen” means fluorine, chlorine, bromine or iodine.
The term“carbonyl” alone or in combination refers to the group -C(O)-.
The term“enantiomer” denotes two stereoisomers of a compound which are non- superimpo sable mirror images of one another.
The term“diastereomer” denotes a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities.
The compounds according to the present invention may exist in the form of their pharmaceutically acceptable salts. The term“pharmaceutically acceptable salt” refers to conventional acid-addition salts or base-addition salts that retain the biological effectiveness and properties of the compounds of formula (I) and are formed from suitable non-toxic organic or inorganic acids or organic or inorganic bases. Acid-addition salts include for example those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid, and those derived from organic acids such as -toluenesulfonic acid, salicylic acid, methane sulfonic acid, oxalic acid, succinic acid, citric acid, malic acid, lactic acid, fumaric acid, and the like. Base-addition salts include those derived from ammonium, potassium, sodium and, quaternary ammonium hydroxides, such as for example, tetramethyl ammonium hydroxide. The chemical modification of a pharmaceutical compound into a salt is a technique well known to pharmaceutical chemists in order to obtain improved physical and chemical stability, hygroscopicity, flowability and solubility of compounds. It is for example described in Bastin R.J., et al., Organic Process Research & Development 2000, 4, 427-435. Particular are the sodium salts of the compounds of formula (I).
Compounds of the general formula (I) which contain one or several chiral centers can either be present as racemates, diastereomeric mixtures, or optically active single isomers. The racemates can be separated according to known methods into the enantiomers. Particularly, diastereomeric salts which can be separated by crystallization are formed from the racemic mixtures by reaction with an optically active acid such as e.g. D- or L-tartaric acid, mandelic acid, malic acid, lactic acid or camphor sulfonic acid. cccDNA INHIBITORS
The present invention provides (i) a compound having the general formula (I):
Figure imgf000006_0001
wherein,
W is O or S;
Ai is CH or CR4;
A2 is CH or CR4;
A3 is CH or CR4;
A4 is N, CH or CR4;
A5 is N, CH or CR4;
A6 is CH;
A7 is N or CH;
R1 is halogen, Ci-6alkyl or C3-7cycloalkyl;
R2 is H, OH, halogen, Ci-6alkoxy, haloCi-6alkoxy or phenylCi-6alkoxy;
R3 is carboxy or Ci-6alkoxycarbonyl; R4 is halogen, OH, CN, Ci-6alkyl, C3-7cycloalkyl, Ci-6alkoxy, oxopyrrolidinyl, morpholinyl or haloCi-6alkyl;
Gi is Ci-6alkyl, hydroxyCi-6alkyl or C3-7cycloalkylCi-6alkyl;
G2 is Ci-6alkyl, C3-7cyclo alkyl or phenyl;
or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
A another embodiment of the present invention is a compound of formula (I), wherein W is O or S;
Ai is CH or CR4;
A2 is CH or CR4;
A3 is CH or CR4;
A4 is N, CH or CR4;
A5 is N, CH or CR4;
A6 is CH;
A7 is N or CH;
R1 is halogen, Ci-6alkyl or C3-7cycloalkyl;
R2 is H, OH, halogen or Ci-6alkoxy;
R3 is carboxy or Ci-6alkoxycarbonyl;
R4 is halogen, OH, CN, Ci-6alkyl, C3-7cyclo alkyl or Ci-6alkoxy;
Gi is Ci-6alkyl, hydroxyCi-6alkyl or C3-7cycloalkylCi-6alkyl;
G2 is Ci-6alkyl, C3-7cyclo alkyl or phenyl;
or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
A further embodiment of the present invention is (ii) a compound of formula (I) according to (i), wherein
W is O or S;
Ai is CH or CR4;
A2 is CH or CR4;
A3 is CH or CR4;
A4 is N, CH or CR4;
A5 is N, CH or CR4;
A6 is CH;
A7 is N or CH;
R1 is Cl, Br, methyl or cyclopropyl;
R2 is H, OH, Cl, Br, methoxy, trifluoromethoxy or benzyloxy; R3 is carboxy, methoxycarbonyl or ethoxycarbonyl;
R4 is F, Cl, Br, OH, CN, methyl, isopropyl, cyclopropyl, methoxy, ethoxy, isopropoxy, oxopyrrolidinyl, morpholinyl or trifluoromethyl;
Gi is cyclobutylmethyl, ethyl, hydroxypropyl, methylethyl or propyl;
G2 is cyclobutyl, cyclohexyl, cyclopentyl, cyclopropyl, dimethylethyl, ethyl, methyl and phenyl;
or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
A further embodiment of the present invention is a compound of formula (I), wherein W is O or S;
Ai is CH or CR4;
A2 is CH or CR4;
A3 is CH or CR4;
A4 is N, CH or CR4;
A5 is N, CH or CR4;
A6 is CH;
A7 is N or CH;
R1 is Cl, Br, methyl or cyclopropyl;
R2 is H, OH, Cl or methoxy;
R3 is carboxy, methoxycarbonyl or ethoxycarbonyl;
R4 is F, Cl, Br, OH, CN, methyl, isopropyl, cyclopropyl, methoxy, ethoxy or isopropoxy; Gi is cyclobutylmethyl, ethyl, hydroxypropyl, methylethyl or propyl;
G2 is cyclobutyl, cyclohexyl, cyclopentyl, cyclopropyl, dimethylethyl, ethyl, methyl and phenyl;
or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
A further embodiment of the present invention is (iii) a compound of formula (I) or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein Ai is CH.
A further embodiment of the present invention is (iv) a compound of formula (I) or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein A4 is CH or CR4.
A further embodiment of the present invention is (v) a compound of formula (I) or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein As is CH or CR4. A further embodiment of the present invention is (vi) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein A7 is CH.
A further embodiment of the present invention is (vii) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R1 is halogen.
A further embodiment of the present invention is (viii) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R2 is H.
A further embodiment of the present invention is (ix) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R3 is carboxy.
A further embodiment of the present invention is (x) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R4 is halogen, Ci-6alkyl or Ci-6alkoxy.
A further embodiment of the present invention is (xi) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein Gi is Ci-6alkyl.
A further embodiment of the present invention is (xii) a compound of formula (I), or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein G2 is Ci-6alkyl or C3-7cycloalkyl.
Another further embodiment of the present invention is (xiii) a compound of formula (I), wherein
W is O or S;
Ai is CH;
A2 is CH or CR4;
A3 is CH or CR4;
A4 is CH or CR4;
A5 is CH or CR4;
A6 is CH;
A7 is CH;
R1 is halogen;
R2 is H;
R3 is carboxy; R4 is halogen, Ci-6alkyl or Ci-6alkoxy;
Gi is Ci-6alkyl;
G2 is Ci-6alkyl or C3-7cyclo alkyl;
or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
Another further embodiment of the present invention is (xiv) a compound of formula (I), wherein
W is O or S;
Ai is CH;
A2 is CH or CR4;
A3 is CH or CR4;
A4 is CH or CR4;
A5 is CH or CR4;
A6 is CH;
A7 is CH;
R1 is Cl, Br;
R2 is H;
R3 is carboxy;
R4 is F, Cl, Br, methyl, ethoxy or methoxy;
Gi is ethyl or methylethyl;
G2 is cyclobutyl or methyl;
or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
In another embodiment (xv) of the present invention, particular compounds of the present invention are selected from:
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
Ethyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylate; 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]benzoic acid;
Ethyl 2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopropanecarboxylate; cis-2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopropanecarboxylic acid; trans-2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopropanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopentanecarboxylic acid;
cA-4-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclohexanecarboxylic acid;
/r<mv-4-[2-[4-(8-chloiO-4-oxo-chiOiTien-2-yl)phenoxy]ethoxy]cyclohexanecarboxylic acid; 2-[3-[[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]methyl]cyclobutoxy]acetic acid;
2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]propanoic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy] -2,2-dimethyl-propanoic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-l-methyl-ethoxy]cyclobutanecarboxylic acid; 3-[3-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]propoxy]cyclobutanecarboxylic acid;
3-[3-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-2-hydroxy-propoxy]benzoic acid;
3-[2-[4-(8-chloro-7-fluoro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-7-methoxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-6-fluoro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(7,8-dichloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-6-cyano-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-7-cyano-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; r/.v-3-[2-[4-(8-chloro-5-hydroxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
r/.v-3-[2-[4-(8-chloro-5-methoxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
c/5,-3-[2-[4-(8-chloro-5-isopropoxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid;
r/.v-3-[2-[4-(5-bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
r/.v-3- [2- [4-(5,8-dichloro-4-oxo-chromen-2-yl)phenoxy]ethoxy] cyclobutanecarboxylic acid; c/5,-3-[2-[4-(8-chloro-5-cyclopropyl-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid;
r/.v-3-[2-[4-(8-chloro-5-methyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
r/.v-3- [2- [4-(8-bromo-4-oxo-chromen-2-yl)phenoxy]ethoxy] cyclobutanecarboxylic acid;
r/.v-3-[2-[4-(8-cyclopropyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; r/.v-3- [2- [4-(8-methyl-4-oxo-chromen-2-yl)phenoxy]ethoxy] cyclobutanecarboxylic acid;
2-[2-[4-(8-chloro-5-fluoro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetic acid;
r/v-3-[2-[[5-(8-chloiO-4-oxo-chiOinen-2-yl)-2-pyridyl]oxy]ethoxy]cyclobutanecarboxylic acid; /r< s,-3-[2-[[5-(8-chloro-4-oxo-chromen-2-yl)-2-pyridyl]oxy]ethoxy]cyclobiitanecarboxylic acid;
r/.v-3-[2-[5-(8-chloro-4-oxo-chromen-2-yl)pyrazin-2-yl]oxyethoxy]cyclobutanecarboxylic acid;
r/'.v- methyl 3-[2-[5-(8-chloro-4-oxo-chromen-2-yl)pyrazin-2- yl] oxyethoxy] cyclobutanecarboxylate;
r/'.v- methyl 3-[2-[6-(8-chloro-4-oxo-chromen-2-yl)pyridazin-3- yl] oxyethoxy] cyclobutanecarboxylate;
r/.v-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-fluoro-phenoxy]ethoxy]cyclobutanecarboxylic acid;
r/.v-3-[2-[2-chloro-4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
,v-3-[2-[2-bromo-4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
,v-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyclopropyl- phenoxy] ethoxy] cyclobutanecarboxy lie acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyano-phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-methyl-phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-isopropyl-phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-methoxy-phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-ethoxy-phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(3,8-dichloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
r/v-3-[2-[4-(8-chloiO-3-hydiOxy-4-oxo-chiOmen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
r/v-3-[2-[4-(8-chloiO-3-methoxy-4-oxo-chiOmen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-4-oxo-thiochromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
2-[2-[4-(8-chloro-4-oxo-thiochromen-2-yl)phenoxy]ethoxy] acetic acid; and
2-[2-[4-(8-chloro-3-hydroxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetic acid;
or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof. In a further embodiment (xvi) of the present invention, more particular compounds of the present invention are selected from:
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
cis-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
trans-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-l-methyl-ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-6-fluoro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; cis-3-[2-[4-(8-chloro-5-methoxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
cis-3-[2-[4-(5,8-dichloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; cis-3-[2-[4-(8-bromo-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
cis-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-fluoro-phenoxy]ethoxy]cyclobutanecarboxylic acid;
cis-3-[2-[2-chloro-4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
cis-3-[2-[2-bromo-4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-methyl-phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-ethoxy-phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-thiochromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
2-[2-[4-(8-chloro-4-oxo-thiochromen-2-yl)phenoxy]ethoxy] acetic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-morpholino-phenoxy]ethoxy]cyclobutanecarboxylic acid;
2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetamide;
Methyl 3-[2-[4-[8-chloro-6-fluoro-4-oxo-3-(trifluoromethoxy)chromen-2- yl]phenoxy] ethoxy] cyclobutanecarboxylate ;
3-[2-[4-[8-chloro-6-fluoro-4-oxo-3-(trifluoromethoxy)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(3-bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(3-benzyloxy-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-[8-chloro-4-oxo-6-(trifluoromethyl)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-6-cyclopropyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-[8-chloro-4-oxo-6-(2-oxopyrrolidin-l-yl)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-7-cyclopropyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
ethyl 2-[2-[4-(7-bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetate;
3-[2-[4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2-yl]-2-methyl- phenoxy] ethoxy] cyclobutanecarboxy lie acid; and
C/5,-3-[2-[2-chloro-4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylic acid; or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
Similar flavone compounds, 6,4’-dimethoxyflavone (compound F-l), 4’-methoxyflavone (compound F-2) disclosed in patent WO2015061294 for treating HBV infection as STING agonist and 4-(7-methoxy-4-oxo-chromen-3-yl)benzoic acid (compound F-3) was disclosed in patent (CN102617536) for treating HBV infection, were chosen as reference compounds in present invention..
Figure imgf000014_0001
(compound F-3)
SYNTHESIS
The compounds of the present invention can be prepared by any conventional means. Suitable processes for synthesizing these compounds as well as their starting materials are provided in the schemes below and in the subsequent examples. All substituents, in particular, R1 to R3, X, Gi, G2, A1 to A7 are defined as below unless otherwise indicated. Furthermore, and unless explicitly otherwise stated, all reactions, reaction conditions, abbreviations and symbols have the meanings well known to a person of ordinary skill in the art.
Scheme 1
Figure imgf000015_0001
wherein Q is halogen, OTs, OTf or OMs; W is O or S.
Condensation of substituted ketone IY with substituted aldehyde V in the presence of a base, such as KOH, in a suitable solvent, such as ethanol, affords a,b-unsaturated carbonyl intermediate VI. Cyclization of intermediate VI in the presence of a suitable Lewis acid such as I2, KI or Nal, in a suitable solvent, such as DMSO, affords flavone derivatives VII.
Demethylation of intermediate VII with a suitable Lewis acid, such as BBn, in a suitable solvent, such as dichloromethane, affords compound of formula VIII. Substitution of compound of formula VIII with compound of formula IX in the presence of a suitable base, such as K2CO3, in a suitable solvent, such as DMF, affords compound of formula X. Substitution of compound of formula X with compound of formula XI in the presence of a suitable base, such as NaH, in a suitable solvent, such as DMSO, affords final compound of formula I. Alternatively, Substitution of compound of formula VIII with compound of formula XII in the presence of a suitable base such as K2CO3 in a suitable solvent such as DMF also affords compound of formula I.
The final compound of formula I can also be prepared by starting with condensation of substituted ketone IV with substituted aldehyde XIII in the presence of a base, such as KOH, in a suitable solvent, such as ethanol, affords a,b-unsaturated carbonyl compound of formula XIV. Cyclization of compound of formula XIV in the presence of a suitable Lewis acid, such L, in a suitable solvent, such as DMSO, affords final compound of formula I.
Scheme 2
Figure imgf000016_0001
wherein W is O or S.
The intermediate X can also be prepared according to the Scheme 2. Condensation of substituted ketone IV with substituted aldehyde XVI in the presence of a base, such as KOH, in a suitable solvent, such as ethanol, affords a,b-unsaturated ketone intermediate XVII.
Cyclization of intermediate XVII in the presence of a suitable Lewis acid, such as I2, in a suitable solvent, such as DMSO, affords intermediates X.
This invention also relates to a process for the preparation of a compound of formula (I) comprising the following step:
(a) substitution of a compound of formula (X),
Figure imgf000017_0001
(X),
with a compound of formula (XI) in the presence of a base;
(b) substitution of a compound of formula (IX),
Figure imgf000017_0002
(IX),
with a compound of formula (XII) in the presence of a base;
(c) cyclization of compound of formula (XIV),
Figure imgf000017_0003
in the presence of a suitable Lewis acid;
wherein R1, R2, R3,W, Ai to A7, Gi and G2 are defined as above;
the base in step (a) can be for example NaH;
the base in step (b) can be for example K2CO3;
the Lewis acid in step (c) can be for example I2. A compound of formula (I) when manufactured according to the above process is also an object of the invention.
PHARMACEUTICAL COMPOSITIONS AND ADMINISTRATION
The invention also relates to a compound of formula (I) for use as therapeutically active substance. Another embodiment provides pharmaceutical compositions or medicaments containing the compounds of the invention and a therapeutically inert carrier, diluent or excipient, as well as methods of using the compounds of the invention to prepare such compositions and medicaments. In one example, compounds of formula (I) may be formulated by mixing at ambient temperature at the appropriate pH, and at the desired degree of purity, with physiologically acceptable carriers, i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form. The pH of the formulation depends mainly on the particular use and the concentration of compound, but preferably ranges anywhere from about 3 to about 8. In one example, a compound of formula (I) is formulated in an acetate buffer, at pH 5. In another embodiment, the compounds of formula (I) are sterile. The compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation or as an aqueous solution.
Compositions are formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. The“effective amount” of the compound to be administered will be governed by such considerations, and is the minimum amount necessary to inhibit cccDNA in HBV patients, consequently lead to the reduction of HBsAg and HBeAg (HBV e antigen) in serum. For example, such amount may be below the amount that is toxic to normal cells, or the mammal as a whole.
In one example, the pharmaceutically effective amount of the compound of the invention administered parenterally per dose will be in the range of about 0.1 to 100 mg/kg, alternatively about 0.1 to 50 mg/kg of patient body weight per day, with the typical initial range of compound used being 0.3 to 15 mg/kg/day. In another embodiment, oral unit dosage forms, such as tablets and capsules, preferably contain from about 25 to about 1000 mg of the compound of the invention. The compounds of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
The compounds of the present invention may be administered in any convenient administrative form, e.g., tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components conventional in pharmaceutical preparations, e.g., diluents, carriers, pH modifiers, sweeteners, bulking agents, and further active agents.
A typical formulation is prepared by mixing a compound of the present invention and a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, e.g., Ansel, Howard C., et al., Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R., et al. Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament).
An example of a suitable oral dosage form is a tablet containing about 25 to 500 mg of the compound of the invention compounded with about 90 to 30 mg anhydrous lactose, about 5 to 40 mg sodium croscarmellose, about 5 to 30 mg polyvinylpyrrolidone (PVP) K30, and about 1 to 10 mg magnesium stearate. The powdered ingredients are first mixed together and then mixed with a solution of the PVP. The resulting composition can be dried, granulated, mixed with the magnesium stearate and compressed to tablet form using conventional equipment. An example of an aerosol formulation can be prepared by dissolving the compound, for example 5 to 400 mg, of the invention in a suitable buffer solution, e.g. a phosphate buffer, adding a tonicifier, e.g. a salt such sodium chloride, if desired. The solution may be filtered, e.g., using a 0.2 micron filter, to remove impurities and contaminants.
An embodiment, therefore, includes a pharmaceutical composition comprising a compound of Formula (I), or pharmaceutically acceptable salt or enantiomer or diastereomer thereof.
In a further embodiment includes a pharmaceutical composition comprising a compound of formula (I), or pharmaceutically acceptable salt or enantiomer or diastereomer thereof, together with a pharmaceutically acceptable carrier or excipient.
Another embodiment includes a pharmaceutical composition comprising a compound of formula (I), or pharmaceutically acceptable salt or enantiomer or diastereomer thereof for use in the treatment of HBV infection.
INDICATIONS AND METHODS OF TREATMENT
The compounds of the invention can inhibit cccDNA and have anti-HBV activity. Accordingly, the compounds of the invention are useful for the treatment or prophylaxis of HBV infection.
The invention relates to the use of a compound of formula (I) for the inhibition of cccDNA.
The invention also relates to the use of a compound of formula (I) for the inhibition of HBeAg.
The invention further relates to the use of a compound of formula (I) for the inhibition of HBsAg.
The invention relates to the use of a compound of formula (I) for the inhibition of HBV
DNA.
The invention relates to the use of a compound of formula (I) for the treatment or prophylaxis of HBV infection.
The use of a compound of formula (I) for the preparation of medicaments useful in the treatment or prophylaxis diseases that are related to HBV infection is an object of the invention.
The invention relates in particular to the use of a compound of formula (I) for the preparation of a medicament for the treatment or prophylaxis of HBV infection.
Another embodiment includes a method for the treatment or prophylaxis of HBV infection, which method comprises administering an effective amount of a compound of Formula (I), or enantiomers, diastereomers, prodrugs or pharmaceutically acceptable salts thereof. BRIEF DESCRIPTION OF THE FIGURE(S)
Figure 1: the result of Example 1 in cccDNA Southern Blot assay, it indicates that Example 1 dose-dependently reduced cccDNA level in HepDESl9 cells.
EXAMPLES
The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of the invention.
Abbreviations used herein are as follows:
ACN: acetonitrile
BBr3: boron tribromide
DMAP: 4-dimethylaminopyridine
DME: dimethoxyethane
DMF: /V,/V-dimethylformamide
EC50: the molar concentration of an inhibitor, which produces 50% of the maximum possible response for that inhibitor
FBS: fetal bovine serum
H2O2: hydrogen peroxide
HPLC: high performance liquid chromatography
hr(s): hour(s)
min: minute
MS (ESI): mass spectroscopy (electron spray ionization)
Ms: methylsulfonyl
NCS: N-chloro succinimide
NMP: /V- methyl-2- pyrrol idone
obsd.: observed
PE: petroleum ether
PPA: polyphosphoric acid
PPh3; triphenylpho sphine
Py: pyridine
rt: room temperature Sphos: 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl
Tf: trifluoromethanesulfonyl
TFA: trifluoro acetic acid
TFAA: trifluoro acetic anhydride
TMS : trimethylsilyl
Ts: -tolylsulfonyl
d: chemical shift
GENERAL EXPERIMENTAL CONDITIONS
Intermediates and final compounds were purified by flash chromatography using one of the following instruments: i) Biotage SP1 system and the Quad 12/25 Cartridge module ii) ISCO combi- flash chromatography instrument. Silica gel Brand and pore size: i) KP-SIL 60 A, particle size: 40-60 pm; ii) CAS registry NO: Silica Gel: 63231-67-4, particle size: 47-60 micron silica gel; iii) ZCX from Qingdao Haiyang Chemical Co., Ltd, pore: 200-300 or 300-400.
Intermediates and final compounds were purified by preparative HPLC on reversed phase column using X Bridge™ Perp Ci8 (5 pm, OBD™ 30 x 100 mm) column or SunFire™ Perp Ci8 (5 pm, OBD™ 30 x 100 mm) column.
LC/MS spectra were obtained using a Waters UPLC-SQD Mass. Standard LC/MS conditions were as follows (running time 3 minutes):
Acidic condition: A: 0.1% formic acid and 1% acetonitrile in H20; B: 0.1% formic acid in acetonitrile;
Basic condition: A: 0.05% NH3-H20 in H20; B: acetonitrile.
Mass spectra (MS): generally only ions which indicate the parent mass are reported, and unless otherwise stated the mass ion quoted is the positive mass ion (M+H)+.
NMR Spectra were obtained using Bruker Avance 400MHz.
All reactions involving air-sensitive reagents were performed under an argon atmosphere. Reagents were used as received from commercial suppliers without further purification unless otherwise noted. PREPARATIVE EXAMPLES
Intermediate 1: methyl 3-(2-hydroxyethoxy)cyclobutanecarboxylate
Figure imgf000023_0001
Int-1
Step 1: Preparation of 2-benzyloxyethoxy(trimethyl)silane
Figure imgf000023_0002
Int-la
To a solution of 2-benzyloxyethanol (20.0 g, 131.4 mmol) and TEA (20.0 g, 197.1 mmol) in dichloro methane (200 mL) cooled at 0 °C was added trimethylsilyl chloride (17.1 g, 157.7 mmol) and the mixture was then stirred at 25 °C for 16 hours. After the reaction was completed, the mixture was concentrated in vacuo and the residue was purified by column chromatography on silica gel (elution with PE:EtOAc=50:l to 10:1) to give the 2- benzyloxyethoxy(trimethyl)silane (25.0 g, 84.9%) as a colorless oil.
Step 2: Preparation of methyl 3-(2-benzyloxyethoxy)cyclobutanecarboxylate
Figure imgf000023_0003
Int-lb
To a solution of 2-benzyloxyethoxy(trimethyl)silane (25.0 g, 111.4 mmol) and methyl 3- oxocyclobutanecarboxylate (CAS #: 4934-99-0, Cat.#: PB01390, from PharmaB lock (Nanjing) R&D Co. Ltd, 15.0 g, 117.0 mmol) in dichloro methane (200 mL) was added trimethylsilyl trifluoromethanesulfonate (12.4 g, 55.7 mmol) dropwise at -78 °C. After addition, the mixture was stirred at -78 °C for additional 1 hour and then to the resulting mixture was added triethylsilane (14.25 g, 122.57 mmol). After addition, the resulting mixture was warmed to room temperature and stirred at room temperature for 1 hour. After the reaction was completed, the mixture was washed with saturated NH4Cl solution, brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with PE/EtOAc=l00: 1-50:1) to give methyl 3-(2- benzyloxyethoxy)cyclobutanecarboxylate (28 g, 95.1%) as a colorless oil. MS obsd. (ESI+) [(M+H)+]: 265.1.
Step 3: Preparation of methyl 3-(2-hydroxyethoxy)cyclobutanecarboxylate
Figure imgf000024_0001
Int-1
To a solution of methyl 3-(2-benzyloxyethoxy)cyclobutanecarboxylate (28.0 g, 105.9 mmol) in MeOH (300.0 mL) was added Pd(OH)2(wet) (1.48 g, 10.6 mmol) at room temperature and the mixture was then hydrogenated under H2 atmosphere at room temperature overnight. After the reaction was completed, the reaction was filtered through silica gel pad and the filtrate was concentrated in vacuo to give 18 g crude methyl 3-(2- hydroxyethoxy)cyclobutanecarboxylate (18 g, 97.6%) as a colorless oil.
Intermediate 2: methyl 3-[2-(p-tolylsulfonyloxy)ethoxy]cyclobutanecarboxylate
Figure imgf000024_0002
Int-2
To a solution of methyl 3-(2-hydroxyethoxy)cyclobutanecarboxylate (5 g, 28.7 mmol) and
DMAP (5.26 g, 43.1 mmol) in dichloromethane (80 mL) was added 4-methylbenzene-l-sulfonyl chloride (6.02 g, 31.6 mmol) at room temperature and the mixture was then stirred at room temperature overnight. After the reaction was completed, the mixture was washed with 1N HC1 (25 mL), water (l5mL), saturated NaHCCh solution, brine and concentrated in vacuo to give the crude methyl 3-[2-(p-tolylsulfonyloxy)ethoxy]cyclobutanecarboxylate (8.1 g, 85.6%) as a colorless oil, which was used in next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 329.2.
Intermediate 3: methyl 3-[2-(4-formylphenoxy)ethoxy]cyclobutanecarboxylate
Figure imgf000025_0001
Int-3
To a solution of 4-hydroxybenzaldehyde (5.0 g, 16.38 mmol) and methyl 3-[2-(p- tolylsulfonyloxy)ethoxy]cyclobutanecarboxylate (5.38 g, 16.38 mmol) in DMF (50 mL) was added K2CO3 (4.53 g, 32.75 mmol). The mixture was stirred at 60 °C for 12 hours. The resulting mixture was then poured into water (200mL) and extracted with EtOAc (200 mL) three times. The combined organic layer was washed with brine, dried over Na2S04 and concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with PE: EtOAc=50: 1-10:1) to give methyl 3-[2-(4-formylphenoxy)ethoxy]cyclobutanecarboxylate (3.6 g 78.99%) as a yellow solid. MS obsd. (ESI+) [(M+H)+]: 279.1.
Intermediate 4: .v-ethyl 3-[2-(4-formylphenoxy)ethoxy]cyclobutanecarboxylate
Figure imgf000025_0002
Int-4
Step 1: Preparation of 2-(4-formylphenoxy)ethyl trifluoromethanesulfonate
Figure imgf000025_0003
To a solution of 4- (2-hydro xyethoxy)benzaldehyde (3 g, 18.1 mmol) and 2,6- dimethylpyridine (3.87 g, 4.21 mL, 36.1 mmol) in dichloromethane ( 40 mL) was added trifluoromethanesulfonic anhydride (9.17 g, 5.33 mL, 32.5 mmol) at -30°C and the mixture was then stirred at 0 °C for 1 hour. The mixture was then washed with 1 N HC1 (20 mL) twice, water (20 mL) twice, brine (20 mL), dried over anhydrous Na2S04 and concentrated in vacuo to give the crude 2-(4-formylphenoxy)ethyl trifluoromethane sulfonate (5.4 g, 100%) , which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 299.1.
Step 2: Preparation of .v-ethyl 3-[2-(4-formylphenoxy)ethoxy]cyclobutanecarboxylate
Figure imgf000026_0001
Int-4
To a solution of ethyl 3-hydroxycyclobutanecarboxylate (CAS #: 17205-02-6, Cat.#:
PBN20120730, from PharmaB lock (Nanjing) R&D Co. Ltd, 2.6g, 18 mmol, the ratio of cis isomer : trans isomer = 10 : 1) in THF (20 mL) was added NaH (793 mg, 19.8 mmol) portion wise at 0 °C and the mixture was then stirred at 0 °C for 30 minutes. Then to the resulting mixture was added the solution of 2-(4-formylphenoxy)ethyl trifluoromethanesulfonate (5.4g, 18.1 mmol, crude) in THF (40 mL) dropwise at 0 °C. After addition, the mixture was poured into ice- water (50 mL) and extracted with dichloromethane (50 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with PE:EtOAc=
100: 1 to 3: 1) to give cA-ethyl 3-[2-(4-formylphenoxy)ethoxy]cyclobutanecarboxylate as a yellow solid (1.7 g, 32 % yield). lH NMR (DMSC )-d6, 400MHz): d ppm 9.78-9.94 (m, 1H), 7.74- 7.91 (m, 2H), 7.07-7.19 (m, 2H), 4.19 (dd, 7=3.85, 5.32 Hz, 2H), 4.02-4.10 (m, 2H), 3.90-4.00 (m, 1H), 3.66 (dd, 7=3.79, 5.26 Hz, 2H), 2.60-2.73 (m, 1H), 2.41-2.48 (m, 2H), 1.93-2.05 (m,
2H), 1.10-1.23 (m, 3H). The trans isomer was not collected in the purification.
Intermediate 5: methyl 3-methylsulfonyloxycyclobutanecarboxylate
Figure imgf000026_0002
Int-5 To a solution of methyl 3-hydro xycyclobutanecarboxylate (1 g, 7.68 mmol) and TEA (1.17 g, 1.61 mL, 11.5 mmol) in dichloromethane (10 mL) was added methane sulfonyl chloride (1.14 g, 778 pL, 9.99 mmol) at 0°C and the mixture was then stirred at room temperature overnight. The mixture was then diluted with dichloromethane (50 mL), the resulting solution was then washed with water (20 mL) twice, saturated NaHCCE (20 mL) twice, brine (20 mL), dried over anhydrous Na2S04 and concentrated in vacuo to give the crude methyl 3- methylsulfonyloxycyclobutanecarboxylate (1.6 g, 100%) as a colorless oil. MS obsd. (ESI+) [(M+H)+]: 209.2.
Example 1 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000027_0001
1
Step 1: Preparation of (E)-l-(3-chloro-2-hydroxy-phenyl)-3-[4-(2- hydroxyethoxy)phenyl]prop-2-en-l-one
Figure imgf000027_0002
la
To a solution of l-(3-chloro-2-hydroxy-phenyl) ethanone (8 g, 46.9 mmol) and 4-(2- hydroxyethoxy) benzaldehyde (7.79 g, 46.9 mmol) in EtOH (150 ml) was added KOH (5.26 g, 93.8 mmol) at room temperature and the mixture was then stirred at l00°C for 3 hours. After the reaction was completed, the resulting mixture was adjusted to pH~4 by 2N HC1 to yield a suspension. The solid was collected by filtration and dried in vacuo to give the crude (£)-l-(3- chloro-2-hydroxy-phenyl)-3-[4-(2-hydroxyethoxy)phenyl]prop-2-en-l-one (14 g, 93.7 %) as a light yellow solid. MS obsd. (ESI+) [(M+H)+]: 329.2.
Step 2: Preparation of 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one
Figure imgf000028_0001
To a solution of (£’)-l-(3-chloro-2-hydroxy-phenyl)-3-[4-(2-hydroxyethoxy)phenyl]prop- 2-en-l-one (14 g, 43.9 mmol) in DMSO (50 mL) was added I2 (557 mg, 2.2 mmol) at room temperature and the mixture was then stirred at l40°C for 3hours. After the reaction was completed, to the mixture was added saturated NaHS03 solution (10 mL) and water (40 mL). The resulting suspension was filtered, the solid was collected and dried in vacuo to give the crude 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one (12 g, 86.3 %) as a yellow solid. MS obsd. (ESL) [(M+H)+]: 317.2.
Step 3: Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000028_0002
1
To a solution of 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one (2 g, 6.31 mmol) in DMSO (20 mL) was added sodium hydride (1.14 g, 28.4 mmol) at room temperature. The mixture was stirred at room temperature for 30 minutes and then to the resulting mixture was added methyl 3-((methylsulfonyl)oxy)cyclobutanecarboxylate (5.26 g, 25.3 mmol) within 1 hour. After addition, the mixture was stirred at room temperature for another 4 hours. After the reaction was completed, the reaction was adjusted to pH~4 by addition of 1N HC1 and diluted with water (60 mL). The resulting mixture was extracted by EtOAc (60 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo. The residue was purified by preparative HPLC to give 3-[2-[4-(8-chloro-4-oxo- chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid (1.67 g, 62.7%) as a white solid. The solid was further purified by supercritical fluid chromatography (SFC) to give r/v-3-[2-[4-(8- chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid (Example 1-A ) and
/r<mv-3-[2-[4-(8-chloiO-4-oxo-chiOiTien-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
(Example 1-B). The configuration of Example 1-A and Example 1-B were determined by NOESY.
Example 1: lH NMR (DMSC )-d6, 400MHz): d ppm 12.17 - 12.07 (m, 1H), 8.12 - 8.05 (m, 2H), 8.00 (d, 7=7.8 Hz, 2H), 7.49 (t, 7=7.8 Hz, 1H), 7.18 (d, 7=9.0 Hz, 2H), 7.07 (s, 1H), 4.23 - 4.15 (m, 2H), 4.06 - 3.90 (m, 1H), 3.70 - 3.64 (m, 2H), 2.64 - 2.54 (m, 1H), 2.48 - 2.36 (m, 2H), 2.21 - 1.96 (m, 2H). MS obsd. (ESE) [(M+H)+]: 415.1.
Example 1-A: lH NMR (DMSO-76, 400MHz): d ppm 12.13 (s, 1H), 8.12 - 8.05 (m, 2H), 8.00 (d, 7=7.8 Hz, 2H), 7.49 (t, 7=7.9 Hz, 1H), 7.20 - 7.15 (m, 2H), 7.07 (s, 1H), 4.22 - 4.16 (m, 2H), 3.99 - 3.88 (m, 1H), 3.67 (dd, 7=3.8, 5.3 Hz, 2H), 2.64 - 2.54 (m, 1H), 2.48 - 2.40 (m, 2H), 2.04 - 1.95 (m, 2H). MS obsd. (ESE) [(M+H)+]: 415.1.
Example 1-B: lH NMR (DMSO-76, 400MHz): d ppm 12.12 (br. s., 1H), 8.09 (d, 7=9.0 Hz, 2H), 8.00 (d, 7=8.1 Hz, 2H), 7.49 (t, 7=7.8 Hz, 1H), 7.18 (d, 7=9.0 Hz, 2H), 7.07 (s, 1H), 4.23 - 4.17 (m, 2H), 3.70 - 3.63 (m, 2H), 2.97 - 2.87 (m, 1H), 2.39 (ddd, 7=3.5, 7.0, 13.2 Hz, 2H), 2.21 - 2.11 (m, 2H). MS obsd. (ESE) [(M+H)+]: 415.1.
Figure imgf000029_0001
Example 1-A Example 1-B
(NOESY correlation observed) (no NOESY correlation observed)
Example 2
Ethyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylate
Figure imgf000030_0001
2
To a mixture of 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (0.4 g, 0.96 mmol) in EtOH (10 mL) was added H2SO4 (0.2 mL) and the mixture was then stirred at 60 °C for 12 hours. After the reaction was completed, the mixture was poured into water (50 mL) and the resulting mixture was extracted with EtOAc (50 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo, the residue was then purified by preparative HPLC to give ethyl 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (305.0 mg, 71.7%) as a yellow solid. 1 H NMR
(DMSO -d6, 400MHz): d ppm 8.15-8.12 (m, 1H), 7.98-7.96 (d, 7=8.0 Hz, 2H), 7.76-7.74 (d, 7=8.0 Hz, 2H), 7.08-7.06 (m, 2H), 6.79 (s, 1H), 4.32-4.14 (m, 5H), 3.78-3.76 (m, 2H), 2.57-2.55 (m, 3H), 2.31-2.29 (m, 2H), 1.3-1.25 (m, 3H). MS obsd. (ESL) [(M+H)+]: 443.1.
Example 3
3- [2- [4-(8-chloro-4-oxo-chromen-2-yl)phenoxy] ethoxy] benzoic acid
Figure imgf000030_0002
3
Step 1: Preparation of (E)-l-(3-chloro-2-hydroxy-phenyl)-3-(4-methoxyphenyl)prop-2-en- 1-one
Figure imgf000031_0001
3a
A mixture of l-(3-chloro-2-hydroxy-phenyl)ethanone (2.5 g, 14.7 mmol), 4- methoxybenzaldehyde (2 g, 14.7 mmol) and KOH (1.64 g, 29.3 mmol) in the EtOH (25 mL) was stirred at l00°C for 3hours. The mixture was then adjusted to PH ~4 by addition of 2N HC1 and the resulting suspension was filtered. The solid was collected and dried in vacuo to give the crude (£)-l-(3-chloro-2-hydroxy-phenyl)-3-(4-methoxyphenyl)prop-2-en-l-one (3.3 g, 78%) as a yellow solid, which was used in the next step directly without further purification. MS obsd.
(ESI+) [(M+H)+]: 289.1.
Step 2: Preparation of 8-chloro-2-(4-methoxyphenyl)chromen-4-one
Figure imgf000031_0002
3b
To a solution of (£’)-l-(3-chloro-2-hydroxy-phenyl)-3-(4-methoxyphenyl)prop-2-en-l-one (5.3 g, 18.4 mmol) in DMSO (60 mL) was added I2 (466 mg, 1.84 mmol) and then the mixture was stirred at 140 °C for 3 hours. After the reaction was completed, the reaction was cooled to room temperature, quenched with saturated NaHSCh solution (10 mL) and diluted with water 100 mL. The resulting suspension was filtered and the solid was collected and dried in vacuo to give the 8-chloro-2-(4-methoxyphenyl)chromen-4-one (5 g, 95 % yield) as a yellow solid, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 287.2. Step 3: Preparation of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one
Figure imgf000032_0001
3c
To a solution of 8-chloro-2-(4-methoxyphenyl)chromen-4-one (5 g, 17.4 mmol) in dichloro methane (40 mL) was added BBn (1 M solution in dichloro methane, 69.8 mL, 69.8 mmol) at room temperature, the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was concentrated in vacuo and the residue was suspended in saturated NH4Cl solution (30 mL). The solid was collected by filtration and dried in vacuo to give the crude 8-chloro-2-(4-hydroxyphenyl)chromen-4-one, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 272.1.
Step 4: Preparation of 2-[4-(2-bromoethoxy)phenyl]-8-chloro-chromen-4-one
Figure imgf000032_0002
3d
To a solution of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one (800.0 mg, 2.94 mmol) and K2CO3 (1.2 g, 8.82 mmol) in DMF (20 mL) was added l,2-dibromoethane (2.76 g, 14.7 mmol) at room temperature and the mixture was then stirred at 80 °C for 12 hours. After the reaction was completed, the mixture was diluted with water (30 mL) and extracted with EtOAc (50 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo to give crude 2-[4-(2-bromoethoxy)phenyl]-8-chloro-chromen-4-one (1.0 g, 89.7%) as a yellow solid, which was used in next step directly without further purification. MS obsd. (ESL) [(M+H)+]: 379.0.
Step 5: Preparation of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] benzoate
Figure imgf000033_0001
3e
To a solution of 2-[4-(2-bromoethoxy)phenyl]-8-chloro-chromen-4-one (100.0 mg, 0.26 mmol) and methyl 3-hydroxybenzoate (48.0 mg, 0.32 mmol) in DMSO (3.0 mL) was added K2CO3 (107.6 mg, 0.78 mmol) at room temperature. Then the mixture was stirred at room temperature under N2 for 10 hours. After the reaction was completed, the mixture was diluted with water (30 mL) and extracted with EtOAc (50 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo to give crude methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]benzoate (117 mg, 100%) as a yellow solid, which was used in next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 451.2.
Step 6: Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]benzoic acid
Figure imgf000033_0002
3 To a solution of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]benzoate
(117.0 mg, 0.26 mmol) in the mixed solvent of MeOH (20 mL) and H20 (3 mL) was added LiOH*H20 (70.0 mg, 1.67 mmol) at room temperature. The mixture was then stirred at room temperature for 48 hours. After the reaction was completed, the reaction was adjusted to pH~4 by addition of 4N HC1. The resulting mixture was then concentrated in vacuo, the residue was purified by preparative HPLC to give 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy]ethoxy]benzoic acid (8.0 mg, 7.0% ) as a white solid. 1 H NMR (DMSO-J^, 400MHz): d ppm 8.07-8.15 (m, 2H), 8.00 (d, J= 7.8 Hz, 2H), 7.39-7.59 (m, 4H), 7.24 (d, 7=8.8 Hz, 3H), 7.08 (s, 1H), 4.45 ppm (dd, 7=18.3, 4.5 Hz, 4H). MS obsd. (ESI+) [(M+H)+]:437.2.
Example 4 Ethyl 2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopropanecarboxylate
Figure imgf000034_0001
Step 1: Preparation of 2-vinyloxy ethyl 4-methylbenzenesulfonate
Figure imgf000034_0002
4a
To a solution of 2-vinyloxyethanol (4.0 g, 45.4 mmol) and TEA (12.66 mL, 90.8 mmol) in dichloromethane (50 mL) was added 4-methylbenzene-l-sulfonyl chloride (12.98 g, 68.1 mmol) at 0 °C and the mixture was then stirred at room temperature for 4 hour. After the reaction was completed, the mixture was concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with petroleum ether: EtOAc = 50:1-5:1) to give 2- vinyloxyethyl 4-methylbenzenesulfonate (10.5 g, 95.46%). MS obsd. (ESI+) [(M+H)+]:243.l. Step 2: Preparation of ethyl 2-[2-(p-tolylsulfonyloxy)ethoxy]cyclopropanecarboxylate
Figure imgf000034_0003
4b To a solution of 2-vinyloxyethyl 4-methylbenzenesulfonate (1.0 g, 4.13 mmol) and rhodium acetate (0.1 g, 0.230 mmol) cooled at 0 °C in dichloro methane (10 mL) was added ethyl 2-diazoacetate (0.57 g, 4.95 mmol) dropwise. After addition, the mixture was stirred at room temperature for 12 hours. After the reaction was completed, the resulting mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with petroleum ether: EtOAc = 50:1-5:1) to give ethyl 2-[2-(p- tolylsulfonyloxy)ethoxy]cyclopropanecarboxylate (1.1 g, 81.16%). MS obsd. (ESI+)
[(M+H)+]:329. l.
Step 3: Preparation of ethyl 2-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy]ethoxy]cyclopropanecarboxylate
Figure imgf000035_0001
4
To a mixture of ethyl 2-[2-(p-tolylsulfonyloxy)ethoxy]cyclopropanecarboxylate (145.72 mg, 0.44 mmol) and 8-chloro-2-(4-hydroxyphenyl)chromen-4-one (110.0 mg, 0.40 mmol) in DMF (5 mL) was added K2CO3 (167.2 mg, 1.2 mmol) at room temperature and the mixture was then stirred at 80 °C for 12 hours. After the reaction was completed, to the mixture was added H20 (lOmL) and the resulting mixture was extracted with ethyl acetate (10 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04, and concentrated in vacuo to give the crude ethyl 2-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclopropanecarboxy late. The crude was further purified by preparative HPLC to give two diastereomers with cis- and trans- configuration, one of which is
characterized as Example 4-A (40 mg, 20.4%) and the other is Example 4-B (60 mg, 35%).
Example 4-A: lH NMR (DMSO -d6, 400MHz): d ppm 8.15 - 8.06 (m, 7=8.9 Hz, 2H), 8.00 (d, 7=7.9 Hz, 2H), 7.49 (t, 7=7.8 Hz, 1H), 7.21 - 7.14 (m, 7=8.9 Hz, 2H), 7.06 (s, 1H), 4.23 (t, 7=4.4 Hz, 2H), 4.05 (dq, 7=1.8, 7.1 Hz, 2H), 3.95 - 3.81 (m, 2H), 3.70 (dt, 7=2.1, 4.4 Hz, 1H), 1.84 - 1.76 (m, 1H), 1.34 - 1.23 (m, 1H), 1.22 - 1.12 (m, 4H). MS obsd. (ESE) [(M+H)+]: 429.1.
Example 4-B: lH NMR (DMSO-76, 400MHz): d ppm 8.09 (d, 7=9.0 Hz, 2H), 8.00 (d, 7=7.8 Hz, 2H), 7.50 (t, 7=7.9 Hz, 1H), 7.16 (d, 7=9.0 Hz, 2H), 7.07 (s, 1H), 4.19 (br d, 7=3.3 Hz, 2H), 4.08 - 4.01 (m, 2H), 3.87 - 3.80 (m, 1H), 3.79 - 3.60 (m, 2H), 1.80 - 1.73 (m, 1H), 1.35 (d, 7=4.8 Hz, 1H), 1.21 - 1.15 (m, 4H). MS obsd. (ESI+) [(M+H)+]: 429.1.
Example 5-A and Example 5-B
Cis-2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopropanecarboxylic acid and trans-2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopropanecarboxylic acid
Figure imgf000036_0001
5-A, 5-B
To a solution of ethyl 2-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclopropanecarboxy late (520.0 mg, 1.2 mmol) in the mixed solvent of THF (2 mL) and water (2 mL) was added LiOFHFEO (96 mg , 2.43 mmol). The mixture was stirred at room temperature for 4 hours. After the reaction was completed, the mixture was adjusted to pH~5 by addition of 1N HC1. The resulting mixture was extracted by EtOAc (20 mL) three times. The combined organic layer was washed with brine, dried with anhydrous Na2S04 and
concentrated in vacuo. The residue was purified by preparative HPLC to give two diastereomers of 2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopropanecarboxylic acid with cis- and trans- configuration, one of which is characterized as Example 5-A (2.9 mg, 3.6%) and the other is Example 5-B (l7mg, 35.3%).
Example 5-A: lH NMR (DMSO -d6, 400MHz): d ppm 8.05 (br d, 7=8.5 Hz, 2H), 7.97 (d, 7=7.9 Hz, 2H), 7.47 (t, 7=7.8 Hz, 1H), 7.15 (br d, 7=8.5 Hz, 2H), 7.02 (s, 1H), 4.20 - 4.11 (m, 2H), 3.86 - 3.70 (m, 2H), 3.65 - 3.56 (m, 1H), 1.59 (q, 7=7.1 Hz, 1H), 1.31 - 1.18 (m, 1H), 0.97 - 0.81 (m, 1H). MS obsd. (ESE) [(M+H)+]: 401.0.
Example 5-B: lH NMR (DMSO-76, 400MHz): d ppm 8.13 (d, 7=8.8 Hz, 2H), 8.04 (d, 7=7.9 Hz, 2H), 7.54 (t, 7=7.8 Hz, 1H), 7.22 (d, 7=8.8 Hz, 2H), 7.10 (s, 1H), 4.27 (br t, 7=4.3 Hz, 2H), 3.99 - 3.85 (m, 2H), 3.63 (br d, 7=4.3 Hz, 1H), 1.68 (br t, 7=6.9 Hz, 1H), 1.15 - 1.07 (m,
2H). MS obsd. (ESE) [(M+H)+]: 401.0.
Example 6 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopentanecarboxylic acid
Figure imgf000037_0001
6
Step 1: Preparation of methyl 3-(2-hydroxyethoxy)cyclopentanecarboxylate
Figure imgf000037_0002
6a
Compound 6a was prepared in analogy to the procedure described for the preparation of Int-1 by using methyl 3-oxocyclopentanecarboxylate as the starting materials instead of methyl 3-oxocyclobutanecarboxylate in Step 2.
Step 2: Preparation of methyl 3-[2-(p-tolylsulfonyloxy)ethoxy]cyclopentanecarboxylate
Figure imgf000037_0003
6b
Compound 6b was prepared in analogy to the procedure described for the preparation of Int-2 by using methyl 3-(2-hydroxyethoxy)cyclopentanecarboxylate as the starting materials instead of methyl 3-(2-hydroxyethoxy)cyclobutanecarboxylate in Step 2.
Step 3: Preparation of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclopentanecarboxy late
Figure imgf000038_0001
6c
To a solution of methyl 3-[2-(p-tolylsulfonyloxy)ethoxy]cyclopentanecarboxylate (1.0 g, 3.67 mmol) and K2CO3 (l.02g, 7.33 mmol) in DMF (20 mL) was added 8-chloro-2-(4- hydroxyphenyl)chromen-4-one (compound 3c, l.38g, 4.03 mmol). The mixture was then stirred at 80 °C for 4 hours. After the reaction was completed, to the mixture was added H20 (50mL) and the resulting mixture was extracted with EtOAc (30 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo to give the crude methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopentanecarboxylate (1.6 g, 98.4%) as a yellow solid, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 443.1.
Step 4: Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclopentanecarboxy lie acid
Figure imgf000038_0002
6
To a solution of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclopentanecarboxy late (1.6 g, 3.61 mmol) in the mixed solvent of THF (10 mL) and H20 (10 mL) was added LiOH*H20 (259.55 mg, 10.84 mmol). The mixture was stirred at room temperature for 4 hours. After the reaction was completed, the mixture was adjusted to pH ~5 by addition of 1N HC1. The resulting mixture was extracted by EtOAc (20 mL) three times. The combined organic layer was washed with brine, dried with anhydrous Na2S04 and concentrated in vacuo to give the crude 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclopentanecarboxy lie acid (1.2 g, 77.7%) as a solid.
Example 6: lH NMR (DMSC )-d6, 400MHz): d ppm 12.07 (br s, 1H), 8.12 - 8.05 (m, 7=8.9 Hz, 2H), 7.99 (d, 7=7.9 Hz, 2H), 7.49 (t, 7=7.8 Hz, 1H), 7.21 - 7.14 (m, 7=8.9 Hz, 2H), 7.06 (s, 1H), 4.25 - 4.13 (m, 2H), 4.09 - 3.88 (m, 1H), 3.76 - 3.66 (m, 2H), 2.80 - 2.78 (m, 1H), 1.99 -
1.78 (m, 4H), 1.75 - 1.61 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 429.1.
Example 7- A and Example 7-B
C7s-4-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclohexanecarboxylic acid and irans-4-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclohexanecarboxylic acid
Figure imgf000039_0001
7-A and 7-B
Step 1: Preparation of methyl 4-(2-hydroxyethoxy)cyclohexanecarboxylate
Figure imgf000039_0002
7a
Compound 7a was prepared in analogy to the procedure described for the preparation of Int-1 by using methyl 4-oxocyclohexanecarboxylate as the starting materials instead of methyl 3-oxocyclobutanecarboxylate in Step 2.
Step 2: Preparation of methyl 4-[2-(p-tolylsulfonyloxy)ethoxy]cyclohexanecarboxylate
Figure imgf000040_0001
7b
Compound 7b was prepared in analogy to the procedure described for the preparation of Int-2 by using methyl 4- (2-hydro xyethoxy)cyclohexanecarboxylate as the starting materials instead of methyl 3-(2-hydroxyethoxy)cyclobutanecarboxylate in Step 2.
Step 3: Preparation of methyl 4-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclohexanecarboxy late
Figure imgf000040_0002
7c
To a solution of methyl 4-[2-(p-tolylsulfonyloxy)ethoxy]cyclohexanecarboxylate (1.0 g,
3.67 mmol) and K2CO3 (l.02g, 7.33 mmol) in DMF (20 mL) was added 2- [4- (2- bromoethoxy)phenyl]-8-chloro-chromen-4-one (l.44g, 4.03 mmol). The mixture was then stirred at 80°C for 4 hours. After the reaction was completed, to the mixture was added H20 (50mL) and the resulting mixture was extracted with EtOAc (30 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo to give the crude methyl 4-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclohexanecarboxylate (l.45g, 86.4%) as a yellow solid, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 457.1.
Step 4: Preparation of 4-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclohexanecarboxy lie acid
Figure imgf000041_0001
7
To a solution of methyl 4-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclohexanecarboxy late (1.45 g, 3.17 mmol) in the mixed solvent of THF (10 mL) and H20 (10 mL) was added LiOH»H20 (259.55 mg, 10.84 mmol). The mixture was stirred at room temperature for 4 hours. After the reaction was completed, the mixture was adjusted to pH~5 by addition of 1N HC1. The resulting mixture was extracted by EtOAc (20 mL) three times. The combined organic layer was with brine, dried with anhydrous Na2S04 and concentrated in vacuo. The residue was purified by preparative HPLC to give two diastereomers of the 4-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclohexanecarboxylic acid with cis- and Irons- configuration, one of which is characterized as Example 7-A (720 mg, 51.2%) and the other is Example 7-B (30mg, 2.1%).
Example 7-A: lH NMR (DMSO -d6, 400MHz): d ppm 12.06 (s, 1H), 8.08 (d, 7=8.9 Hz, 2H), 8.00 (d, 7=7.9 Hz, 2H), 7.49 (t, 7=7.9 Hz, 1H), 7.18 (dd, 7=2.8, 9.0 Hz, 2H), 7.06 (s, 1H), 4.26 - 4.17 (m, 2H), 3.78 (td, 7=4.5, 16.4 Hz, 2H), 3.31 - 3.24 (m, 1H), 2.22 - 2.10 (m, 1H), 2.01
(br d, 7=8.9 Hz, 2H), 1.98 (br d, 7=13.1 Hz, 2H), 1.39-1.37 (m, 2H), 1.22 - 1.18 (m, 2H). MS obsd. (ESI+) [(M+H)+] : 443.0.
Example 7-B: lH NMR (DMSO -d6, 400MHz): d ppm 12.06 (s, 1H), 8.08 (d, 7=8.9 Hz, 2H), 8.00 (d, 7=7.9 Hz, 2H), 7.49 (t, 7=7.9 Hz, 1H), 7.18 (dd, 7=2.8, 9.0 Hz, 2H), 7.06 (s, 1H), 4.26 - 4.17 (m, 2H), 3.78 (td, 7=4.5, 16.4 Hz, 2H), 3.59 - 3.51 (m, 1H), 2.29 - 2.27 (m, 1H),
1.68-1.74 (m, 4H), 1.40 - 1.60 (m, 4H). MS obsd. (ESE) [(M+H)+]: 443.0.
Example 8
2-[3-[[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]methyl]cyclobutoxy]acetic acid
Figure imgf000042_0001
8
Step 1: Preparation of 3-(hydroxymethyl)cyclobutanol
Figure imgf000042_0002
8a
The mixture of methyl 3-oxocyclobutanecarboxylate (3.0 g, 26.3 mmol) in THF (20.0 mL) at -78 °C was added EFHe’MciS (2.0 mL) at room temperature and the mixture was then stirred at room temperature for 12 hours. After the reaction was completed, the mixture was
concentrated in vacuo, the residue was partitioned between dichloro methane (40 mL) and water (50 mL). The mixture was concentrated and extracted with dichloromethane (40.0 mL) and water
(50.0 mL). The organic layer was then separated out and the aquatic phase was extracted with dichloromethane (50 mL) twice. The combined organic layer was washed with brine, dried over Na2S04 and concentrated in vacuo to yield the crude 3-(hydroxymethyl)cyclobutanol (2.0g, 74.5%) as an oil.
Step 2: Preparation of (3-hydroxycyclobutyl)methyl 4-methylbenzenesulfonate
Figure imgf000042_0003
8b To a solution of 3-(hydroxymethyl)cyclobutanol (2.0 g, 20.0 mmol) and TEA (2.0 g, 20.0 mmol) in dichloro methane (50 mL) was added 4-methylbenzene-l-sulfonyl chloride (4.8g, 20.0 mmol) at room temperature and the mixture was then stirred at room temperature for 5 hours. After the reaction was completed, the mixture was washed with 1N HC1 (25 mL), water (l5mL), saturated NaHCCh solution, brine and concentrated in vacuo to give the crude (3- hydroxycyclobutyl)methyl 4-methylbenzenesulfonate (1.0 g, 19.4%) as colorless oil, which was used in next step directly without further purification. MS obsd. (ESI+) [(M+H)+]:257.l.
Step 3: Preparation of 8-chloro-2-[4-[(3-hydroxycyclobutyl)methoxy]phenyl]chromen-4- one
Figure imgf000043_0001
8c
To a solution of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one (1.0 g, 3.7 mmol) and (3- hydroxycyclobutyl)methyl 4-methylbenzenesulfonate (1.0 g, 5.0 mmol) in DMF (10 mL) was added K2CO3 (1.0 g, 10.0 mmol) and the mixture was then stirred at 80 °C for 12 hours. After the reaction was completed, the mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL) three times. The combined organic layer was washed with brine, dried over Na2S04 and concentrated in vacuo to give the crude 8-chloro-2-[4-[(3- hydroxycyclobutyl)methoxy]phenyl]chromen-4-one (1.0 g, 75.6%) as a yellow solid, which was used in next step directly without further purification. MS obsd. (ESI+) [(M+H)+]:357.l.
Step 4: Preparation of ethyl 2-[3-[[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] methyl] cyclobutoxy ] acetate
Figure imgf000043_0002
8d
To a mixture of 8-chloro-2-[4-[(3-hydroxycyclobutyl)methoxy]phenyl]chromen-4-one (0.1 g, 0.3 mmol) and ethyl 2-diazoacetate (0.11 g, 1.0 mmol) in THF (10 mL) at 0 °C was added BF3.Et20 (0.1 mL) and the mixture was then stirred at room temperature for 2 hours. After the reaction was completed, the reaction was quenched with 1N HC1 and extracted with EtOAc (50 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo to give the crude ethyl 2-[3-[[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] methyl] cyclobutoxy] acetate (0.1 g, 75.2%) as a yellow solid, which was used in next step directly without further purification. MS obsd. (ESI+) [(M+H)+]:443. l.
Step 5: Preparation of 2-[3-[[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] methyl] cyclobutoxy] acetic acid
Figure imgf000044_0001
8
The solution of ethyl 2-[3-[[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] methyl] cyclobutoxy] acetate (0.1 g, 0.26 mmol, crude) in the mixed solvent of THF (10 mL) and H20 (10 mL) was added LiOH*H20 (259.55 mg, 10.84 mmol). The mixture was stirred at room temperature for 4 hours. After the reaction was completed, the mixture was adjusted to pH~5 by addition of 1N HC1. The resulting mixture was extracted by EtOAc (20 mL) three times. The combined organic layer was with brine, dried with anhydrous Na2S04 and concentrated in vacuo. The residue was purified by preparative HPLC to give 2-[3-[[4-(8-chloro- 4-oxo-chromen-2-yl)phenoxy]methyl]cyclobutoxy]acetic acid (45.0 mg, 41.7%) as a light yellow solid.
Figure imgf000044_0002
ppm 8.08 (d, 7=8.9 Hz, 2H), 8.00 (d, 7=7.9 Hz, 2H), 7.50 (t, 7=7.8 Hz, 1H), 7.12-7.22 (m, 2H), 7.06 (s, 1H), 4.05-4.13 (m, 2H), 3.89-4.03 (m, 3H), 2.59-2.65 (m, 1H), 2.35-2.41 (m, 1H), 2.20-2.30 (m, 1H), 2.10-2.15 (m, 1H), 1.71-1.83 (m, 1H). MS obsd. (ESL) [(M+H)+]:4l5.2.
Example 9 2- [2- [4-(8-chloro-4-oxo-chromen-2-yl)phenoxy] ethoxy]acetic acid
Figure imgf000045_0001
9
Step 1: Preparation of methyl 2-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] acetate
Figure imgf000045_0002
To the solution of 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one (0.3 g, 0.1 mmol) in DMF (2.5 mL) was added NaH (0.1 g, 60% in oil) and the mixture was stirred at room temperature for 1 hour. Then to the resulting mixture was added methyl 2-chloroacetate (0.3 g, 2.84 mmol) and the mixture was stirred at room temperature for additional 3 hours. After the reaction was completed, the reaction was quenched with water (50 mL) and extracted with dichloromethane (40 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo to give the crude methyl 2-[2-[4-(8-chloro-4- oxo-chromen-2-yl)phenoxy]ethoxy]acetate (0.38g, 100%) as yellow solid, which was used in the next step without further purification. MS obsd. (ESI+) [(M+H)+]:389.2.
Step 2: Preparation of 2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetic acid
Figure imgf000045_0003
9 To a solution of methyl 2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetate (0.38 g, 0.1 mmol, crude) in the mixed solvent of THF (2 mL) and water (2 mL) was added LiOFHFTO (96 mg , 2.43 mmol). The mixture was stirred at room temperature for 4 hours. After the reaction was completed, the mixture was adjusted to pH~5 by addition of 1N HC1. The resulting mixture was extracted by EtOAc (20 mL) three times. The combined organic layer was washed with brine, dried with anhydrous Na2S04 and concentrated in vacuo. The residue was purified by preparative HPLC to give 2-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] acetic acid (153 mg, 40.8%) as a yellow solid. 1 H NMR (DMSO-J^,
400MHz): d ppm 8.10-8.08 (d, J= 8.0 Hz, 2H), 8.00-7.98 (d, J= 8.0 Hz, 2H), 7.51-7.47 (t, 7=7.8 Hz, 1H), 7.19-7.17 (d, 7=8.0 Hz, 2H), 7.06 (s, 1H), 4.25-4.23 (m, 2H), 4.11 (s, 2H), 3.87-3.85 (m, 2H). MS obsd. (ESE) [(M+H)+]:375.0.
Example 10
3- [2- [4-(8-chloro-4-oxo-chromen-2-yl)phenoxy] ethoxy] propanoic acid
Figure imgf000046_0001
10
Example 10 was prepared in analogy to the procedure described for the preparation of example 9 by using ethyl 3-chloropropanoate as the starting material instead of methyl 2- chloro acetate in Step 1.
Example 10:
Figure imgf000046_0002
ppm 8.10-8.08 (d, 7=9.0 Hz, 2H), 8.01- 7.99 (d, 7=8.0 Hz, 2H), 7.52-7.48 (t, 7=7.8 Hz, 1H), 7.19-7.17 (d, 7=8.0 Hz, 2H), 7.07 (s, 1H),
4.25 (s, 2H), 3.77 (s, 2H), 3.72-3.69 (m, 2H), 2.48-2.47 (d, 7=4.0 Hz, 2H). MS obsd. (ESE)
[(M+H)+]: 389.0.
Example 11
3- [2- [4-(8-chloro-4-oxo-chromen-2-yl)phenoxy] ethoxy] -2,2-dimethyl-propanoic acid
Figure imgf000047_0001
11
To a solution of 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one (100.0 mg, 0.32 mmol) and 3-chloro-2, 2-dimethyl-propanoic acid (172.5 mg, 1.26 mmol) in DMF (2 mL) was added NaH (75.8 mg, 1.89 mmol) at room temperature and the mixture was stirred at 80 °C for 16 hours. Then the mixture was poured into water (20 mL) and adjusted to pH~4 by addition of cone. HC1. The resulting mixture was extracted with EtOAc (30 mL) three times. The combined organic layer was washed with brine, dried over Na2S04 and concentrated in vacuo. The residue was then purified by preparative HPLC to give 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] -2,2-dimethyl-propanoic acid (5 mg, 3.7%) as a white solid. 1 H NMR (DMSO-<fc, 400MHZ): d ppm 8.08-8.05 (m, 3H), 7.91-7.89 (m, 1H), 7.48-7.44 (m, 1H), 7.16- 7.14 (d, 7=8.8 Hz, 2H), 6.89 (s, 1H), 4.50-4.48 (m, 2H), 4.36-4.34 (m, 2H), 3.59 (s, 2H), 1.19 (s, 6H). MS obsd. (ESI+) [(M+H)+]: 417.1.
Example 12 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-l-methyl-ethoxy]cyclobutanecarboxylic acid
Figure imgf000047_0002
Step 1: Preparation of (2-chloro-l-methyl-ethoxy)-trimethyl-silane
Figure imgf000048_0001
12a
To a solution of l-chloropropan-2-ol (6.0 g, 63.47 mmol) and TEA (13.27 mL, 95.2 mmol) in dichloro methane (50 mL) was added trimethylsilyl chloride (7.51 g, 69.81 mmol) at 0 °C and the mixture was then stirred at 0 °C for 4 hours. After the reaction was completed, the mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with PE:EtOAc=50:l to 10:1) to give the (2-chloro-l- methyl-ethoxy)-trimethyl-silane (7.2 g, 68.0%) as colorless oil.
Step 2: Preparation of methyl 3-(2-chloro-l-methyl-ethoxy)cyclobutanecarboxylate
Figure imgf000048_0002
12b
To a solution of (2-chloro-l-methyl-ethoxy)-trimethyl-silane (7.2 g, 43.2 mmol) and methyl 3-oxocyclobutanecarboxylate (5.8 g, 45.4 mmol) in dichloro methane (100 mL) was added trimethylsilyl trifluoromethanesulfonate (4.8 g, 21.6 mmol) at -78 °C. After addition, the mixture was stirred at -78 °C for another 1 hour and then to the resulting mixture was added triethylsilane (14.25 g, 122.57 mmol). After addition, the resulting mixture was warmed to room temperature and stirred for 1 hour. After the reaction was completed, the mixture was washed with saturated NH4Cl solution, brine, dried over anhydrous Na2S04 and concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with
PE/EtOAc=l00: 1-50:1) to give methyl 3-(2-chloro-l-methyl-ethoxy)cyclobutanecarboxylate
(5.6 g, 62.7% yield). MS obsd. (ESE) [(M+H)+]: 207.2.
Step 3: Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-l-methyl- ethoxy ] cyclobutanecarboxylic acid
Figure imgf000049_0002
12
Example 12 was prepared in analogy to the procedure described for the preparation of Example 7 by using methyl 3-(2-chloro-l-methyl-ethoxy)cyclobutanecarboxylate as the starting material instead of methyl 4-[2-(p-tolylsulfonyloxy)ethoxy]cyclohexanecarboxylate in Step 3. ln NMR (DMSO-^6, 400MHZ): d ppm 12.18 (br s, 1H), 8.12 - 8.06 (m, J= 8.9 Hz, 2H), 8.00 (d, J= 7.9 Hz, 2H), 7.50 (t, J= 7.9 Hz, 1H), 7.21 - 7.14 (m, 7=8.9 Hz, 2H), 7.07 (s, 1H), 4.07 - 3.97 (m, 3H), 3.87 - 3.74 (m, 1H), , 2.94 - 2.82 (m, 0.5 H), 2.58 - 2.55 (m, 0.5 H), 2.48 - 2.38 (m, 2H),
2.22 - 2.07 (m, 1H), 1.99-1.94 (m, 1H), 1.19 (d, 7=6.2 Hz, 3H). MS obsd. (ESI+) [(M+H)+]:
429.2.
Example 13
3-[3-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]propoxy]cyclobutanecarboxylic acid
Figure imgf000049_0003
13 Step 1: Preparation of 8-chloro-2-[4-(3-hydroxypropoxy)phenyl]chromen-4-one
Figure imgf000049_0001
13a To a solution of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one (400.0 mg, 1.3 mmol) and K2CO3 (1.0 g, 7.2 mmol) in DMF (20 mL) was added 3-bromopropan-l-ol (500 mg, l.3mmol) at room temperature and the mixture was then stirred at 80 °C for 12 hours. After the reaction was completed, the mixture was diluted with water (30 mL) and extracted with EtOAc (50 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo to give the crude 8-chloro-2-[4-(3-hydroxypropoxy)phenyl]chromen-4- one (400 mg, 93.1%) as a yellow solid, which was used in next step directly without further purification. MS obsd. (ESI+) [(M+H)+] : 330.1.
Step 2: Preparation of 3-[3-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy]propoxy]cyclobutanecarboxylic acid
Figure imgf000050_0001
13
Example 13 was prepared in analogy to the procedure described for the preparation of Example 1 by using 8-chloro-2-[4-(3-hydroxypropoxy)phenyl]chromen-4-one as the starting material instead of 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one in Step 3.
Example 13:1H NMR DMSO-d , 400MHz): d ppm 8.08-8.06 (d, J= 6.0 Hz, 2H), 8.00- 7.97 (d, 7=9.0 Hz, 2H), 7.51-7.46 (t, 7=5.8 Hz, 1H), 7.17-7.14 (d, 7=9.0 Hz, 2H), 7.04 (s, 1H), 4.15-4.11 (m, 2H), 3.88-3.83 (m, 1H), 3.54-3.50 (m, 2H), 2.59-2.56 (m, 1H), 2.78-2.65 (m, 2H), 2.03-1.98 (m, 4H). MS obsd. (ESE) [(M+H)+]: 428.9.
Example 14
3-[3-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-2-hydroxy-propoxy]benzoic acid
Figure imgf000050_0002
14 Step 1: Preparation of 8-chloro-2-[4-(oxiran-2-ylmethoxy)phenyl]chromen-4-one
Figure imgf000051_0001
To a mixture of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one (800 mg, 2.93 mmol) and 2- (chloromethyl)oxirane (814 mg, 8.8 mmol) in DMF (20 mL) was added K2CO3 (405 mg, 2.93 mmol) at room temperature and the mixture was then was stirred at room temperature overnight. After the reaction was completed, to the mixture was added water (100 mL), the resulting suspension was filtered. The solid was collected and further purified by recrystallization (MeOH, 20 mL) to give 8-chloro-2-[4-(oxiran-2-ylmethoxy)phenyl]chromen-4-one (650 mg, 67.4%) as a yellow solid. MS obsd. (ESP) [(M+H)+]: 329.2.
Step 2: Preparation of methyl 3-[3-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-2-hydroxy- propoxy ] benzoate
Figure imgf000051_0002
14b To a solution of 8-chloro-2-[4-(oxiran-2-ylmethoxy)phenyl]chromen-4-one (100 mg, 0.3 mmol), methyl 3-hydroxybenzoate (46.3 mg, 0.3 m mol) in THF (5 mL) was added NaH (40 mg, 60% in mineral oil, 1 m mol), the mixture was then stirred at room temperature overnight. After the reaction was completed, the mixture was concentrated in vacuo to give the crude methyl 3- [3-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-2-hydroxy-propoxy]benzoate (146 mg, 99.8%), which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 481.3. Step 3: Preparation of 3-[3-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-2-hydroxy- propoxy] benzoic acid
Figure imgf000052_0001
14 To a solution of methyl 3-[3-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-2-hydroxy- propoxy]benzoate (146 mg, 0.3 mmol, crude, prepared above) in the mixed solvent of MeOH (5 mL), THF (5 mL) and water (1 mL) was added LiOH (10 mg, 0.4 m mol). The mixture was then stirred at room temperature overnight. After the reaction was completed, to the mixture was added AcOH (60 mg, 1 mmol) and the resulting mixture was then concentrated in vacuo. The residue was then purified by preparative HPLC to give 3-[3-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy]-2-hydroxy-propoxy]benzoic acid (40 mg, 27.9%) as a white solid. 1 H NMR
DMSO-d , 400MHZ): d ppm 12.95-13.03 (m, 1H), 8.06-8.13 (m, 2H), 8.00 (d, 7=8.07 Hz, 2H), 7.47-7.57 (m, 3H), 7.39-7.44 (m, 1H), 7.18-7.26 (m, 3H), 7.05-7.08 (m, 1H), 5.50 (d, 7=4.89 Hz, 1H), 4.20-4.26 (m, 2H), 4.14-4.20 (m, 2H), 4.07-4.13 (m, 1H). MS obsd. (ESI+) [(M+H)+]:
467.2.
Example 15
3-[2-[4-(8-chloro-7-fluoro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000052_0002
15 Step 1: Preparation of (2-chloro-3-fluoro-phenyl) acetate
Figure imgf000053_0001
15a
To a mixture of 2-chloro-3-fluoro-phenol (10.0 g, 68.24 mmol) and TEA (7.6 g, 75.06 mmol) in dichloro methane (150 mL) was added acetyl chloride (5.36 g, 68.24 mmol) at 0 °C and the mixture was then stirred at room temperature for 16 hours. After the reaction was completed, the mixture was poured into water (30mL) and extracted with dichloromethane (50mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo. The residue was then purified by column chromatography on silica gel (elution with PE: EtOAc=50: 1-20:1) to give (2-chloro-3-fluoro-phenyl) acetate (10.0 g, 75%) as colorless oil. MS obsd. (ESE) [(M+H)+]: 189.2.
Step 2: Preparation of l-(3-chloro-4-fluoro-2-hydroxy-phenyl)ethanone
Figure imgf000053_0002
15b
A mixture of (2-chloro-3-fluoro-phenyl) acetate (10.0 g, 53.03 mmol) and AlCE (7.07 g, 53.03 mmol) was stirred at 150 °C for 5 hours. After the reaction was completed, the mixture was poured into water (lOOmL) and extracted with EtOAc (250mL) twice. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo. The residue was then purified by column chromatography on silica gel (elution with PE:
EtOAc=50: 1-20:1) to give l-(3-chloro-4-fluoro-2-hydroxy-phenyl)ethanone (3.0 g, 30.0%) as a white solid. MS obsd. (ESE) [(M+H)+]: 189.2.
Step 3: Preparation of (/i)-l-(3-chloro-4-fluoro-2-hydroxy-phenyl)-3-(4- methoxyphenyl)prop-2-en-l-one
Figure imgf000054_0001
15c
To a solution of l-(3-chloro-4-fluoro-2-hydroxy-phenyl)ethanone (2.0 g, 10.61 mmol) and 4-methoxybenzaldehyde (1.29 mL, 10.61 mmol) in EtOH (30 mL) was added KOH (1.79 g, 31.82 mmol) at room temperature and the mixture was then stirred at l00°C for 3hours. After the reaction was completed, the resulting mixture was adjusted to pH~4 by 2N HC1 to yield a suspension. The solid was collected by filtration and dried in vacuo to give the crude (£)-l-(3- chloro-4-fluoro-2-hydroxy-phenyl)-3-(4-methoxyphenyl)prop-2-en-l-one (1.5 g, 46.1%) as a yellow solid, which was used in the next step directly without further purification. MS obsd. (ESL) [(M+H)+]: 307.0.
Step 4: Preparation of 8-chloro-7-fluoro-2-(4-methoxyphenyl)chromen-4-one
Figure imgf000054_0002
To a solution of (£’)-l-(3-chloro-4-fluoro-2-hydroxy-phenyl)-3-(4-methoxyphenyl)prop-2- en-l-one (1.5 g, 4.9 mmol) in DMSO (30 mL) was added I2 (60 mg, 0.24 mmol) and then the mixture was stirred at 140 °C for 3 hours. After the reaction was completed, the reaction was cooled to room temperature, quenched with saturated NaHSCh solution (10 mL) and diluted with water 100 mL. The resulting suspension was filtered and the solid was collected and further purified by recrystallization (EtOH, 10 mL) to give 8-chloro-7-fluoro-2-(4- methoxyphenyl)chromen-4-one (1.2 g, 80.3%) as a yellow solid. MS obsd. (ESI+) [(M+H)+]: 304.9.
Step 4: Preparation of 8-chloro-7-fluoro-2-(4-hydroxyphenyl)chromen-4-one
Figure imgf000055_0001
15e
To a solution of 8-chloro-7-fluoro-2-(4-methoxyphenyl)chromen-4-one (1.2 g, 39 mmol) in dichloro methane (20 mL) was added BBn (2.96 g, 11.81 mmol) at room temperature and then stirred overnight. After the reaction was completed, the reaction was quenched by adding into saturated NaHC03 solution (250 mL) slowly. The resulting suspension was filtered, the solid was collected and dried in vacuo to give the crude 8-chloro-7-fluoro-2-(4- hydroxyphenyl)chromen-4-one (700 mg, 61.8%) as a yellow solid, which was used in the next step directly without further purification. (ESI+) [(M+H)+]: 290.9.
Step 5: Preparation of methyl 3-[2-[4-(8-chloro-7-fluoro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxylate
Figure imgf000055_0002
15f
To a solution of 8-chloro-7-fluoro-2-(4-hydroxyphenyl)chromen-4-one (350 mg, 1.2 mmol) and methyl 3-[2-(p-tolylsulfonyloxy)ethoxy]cyclobutanecarboxylate (435 mg, 1.3 mmol) in
DMF (10 mL) was added K2CO3 (333 mg, 2.4mmol) in DMF (20 mL) at room temperature. The mixture was then stirred at 80 °C for 4 hours. After the reaction was completed, to the mixture was added FhO (50mL) and the resulting mixture was extracted with EtOAc (30 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and
concentrated in vacuo to give the crude methyl 3-[2-[4-(8-chloro-7-fluoro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (200 mg, 37.2%) as a yellow solid, which was used in the next step directly without further purification. (ESI+) [(M+H)+]: 447.0. Step 6: Preparation of 3-[2-[4-(8-chloro-7-fluoro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000056_0001
15 A mixture of methyl 3-[2-[4-(8-chloro-7-fluoro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (200 mg, 0.45 mmol) in con. HC1 (5 mL) was stirred at l00°C for 4 hours. After the reaction was completed, the mixture was concentrated in vacuo. The residue was purified by preparative HPLC to give the 3-[2-[4-(8-chloro-7-fluoro-4-oxo- chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid (50 mg, 25.7%) as a yellow solid. 1 H NMR (DMSO-rfc, 400MHz): d ppm 12.18 (s, 1H), 8.06-8.01 (m, 3H), 7.57-7.55 (m, 1H), 7.16- 7.04 (m, 2H), 7.04 (s, 1H), 4.20-4.18 (d, 7=4.0 Hz, 2H), 3.97-3.95 (m, 1H), 3.68-3.66 (m, 2H), 2.92-2.87 (m, 0.4 H), 2.59-2.57 (m, 1H), 2.46-2.42 (m, 2 H), 2.21-2.05 (m, 0.6 H), 2.03-1.96 (m, 1H). MS obsd. (ESI+) [(M+H)+]: 433.0.
Example 19
3-[2-[4-(8-chloro-7-methoxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000056_0002
19 Step 1: Preparation of l-(3-chloro-2,4-dihydroxy-phenyl)ethanone
Figure imgf000057_0001
19a
To a mixture of l-(2,4-dihydroxyphenyl)ethanone (10 g, 62.7mmol) in H20 (350 mL) was added NaOH (3.2 g, 78.9 mmol) at room temperature. Then to the resulting mixture was added NaClO (5.38 g, 72.3 mmol) slowly (over 1 hour) while keeping the inner temperature no more than 20°C. After addition, the mixture was stirred at room temperature for 17 hour. Then the mixture was adjusted to pH~3 by addition of 1N HC1 solution. The resulting mixture was extracted with EtOAc (400 mL) three times. The combined organic layer was washed with saturated Na2S203 (200 mL) solution twice, water (300 mL), brine (200 mL), dried over Na2S04 and concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with PE:EtOAc =20:1-5:1) to give l-(3-chloro-2,4-dihydroxy-phenyl)ethanone (3g, 25.6% ) as a solid. MS obsd. (ESL) [(M+H)+]: 187.0.
Step 2: Preparation of 3-[2-[4-[(£ 3-(3-chloro-2,4-dihydroxy-phenyl)-3-oxo-prop-l- enyl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000057_0002
To a solution of l-(3-chloro-2,4-dihydroxy-phenyl)ethanone (630 mg, 3.4 mmol) and methyl 3-[2-(4-formylphenoxy)ethoxy]cyclobutanecarboxylate (int-3, l.03g, 3.71 mmol) in EtOH (10 mL) was added KOH (0.57 g, 10.1 mmol) at room temperature and the mixture was then stirred at 80°C for 24hours. After the reaction was completed, the mixture was poured into water (20 mL) and adjusted to pH~4 by 2N HC1, the resulting mixture was extracted with EtOAc (30 mL) three times. The combined organic layer was washed with brine, dried over Na2S04 and concentrated in vacuo to give the crude 3-[2-[4-[(£)-3-(3-chloro-2,4-dihydroxy-phenyl)-3-oxo- prop-l-enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid (200 mg, 13.5%) as a brown oil, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 433.1.
Step 3: Preparation of 3-[2-[4-(8-chloro-7-hydroxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000058_0001
19c
To a solution of 3-[2-[4-[(£)-3-(3-chloro-2,4-dihydroxy-phenyl)-3-oxo-prop-l- enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid (200 mg, 0.46 mmol, crude) in DMSO (3 mL) was added I2 (6 mg, 0.02mmol) at room temperature and then the mixture was stirred at 140 °C for 3 hours. After the reaction was completed, the reaction was cooled to room temperature, quenched with saturated NaHSCh solution (20 mL) and then extracted with EtOAc (20 mL) three times. The combined organic layer was washed with brine, dried over Na2S04 and concentrated in vacuo to give the crude 3-[2-[4-(8-chloro-7-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid as a brown solid. The crude was further purified by recrystallization (EtOAc, 5mL) to give 3-[2-[4-(8-chloro-7-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (60 mg, 30.2%) as a grey solid. 1 H NMR
(DMSO-^6, 400MHZ): d ppm 8.07-8.05 (d, 7=8.8, 2H), 7.85-7.83 (d, 7=8.8, 1H), 7.18-7.16 (m, 2H), 7.12-7.10 (m, 1H), 6.93 (s, 1H), 4.20-4.18 (m, 2H), 3.99-3.91 (m, 1H), 3.68-3.66 (m, 2H), 2.63-2.59 (m, 1H), 2.42-2.41 (m, 2H), 2.03-1.96 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 431.0.
Step 4: Preparation of methyl 3-[2-[4-(8-chloro-7-methoxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy late
Figure imgf000059_0001
19d
To a mixture of 3-[2-[4-(8-chloro-7-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (50.0 mg, 0.120 mmol) and K2CO3 (48.12 mg, 0.350 mmol) in DMF (2 mL) was added Mel (41.18 mg, 0.290 mmol) at room temperature. The mixture was then stirred at room temperature for 16 hours. After the reaction was completed, the mixture was poured into water (5.0 mL) and the resulting suspension was filtered. The solid was collected and dried in vacuo to give the crude methyl 3-[2-[4-(8-chloro-7-methoxy-4-oxo- chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylate (50.0 mg, 90.7%) as a yellow solid, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 459.1.
Step 5: Preparation of 3-[2-[4-(8-chloro-7-methoxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxylic acid
Figure imgf000059_0002
19
To a mixture of methyl 3-[2-[4-(8-chloro-7-methoxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (50.0 mg, 0.110 mmol) in the mixed solvent of THF (2 mL) and water (1 mL) was added LiOH*H20 (13.72 mg, 0.330 mmol) at room temperature. Then the mixture was stirred at room temperature for 16 hours. After the reaction was completed, the mixture was poured into water (5.0 mL) and resulting suspension was filtered. The solid was collected and dried in vacuo to give 3-[2-[4-(8-chloro-7-methoxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (20 mg, 40.8%) as a yellow solid. 1 H NMR (DMSO-^6, 400MHZ): d ppm 8.08-8.06 (d, J= 8.8, 2H), 8.01-7.98 (d, J= 8.8 Hz, 1H), 7.38-7.36 (d, 7=8.8 Hz, 1H), 7.18-7.16 (d, 7=8.8 Hz, 2H), 6.96 (s, 1H), 4.20-4.17 (m, 2H), 4.03 (s, 3H), 3.97- 3.93 (m, 1H), 3.68-3.66 (m, 2H), 2.61-2.47 (m, 0.6 H),2.46-2.43(m, 2H), 2.41-2.03 (m, 0.4H), 2.00-1.98 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 445.1.
Example 20
3-[2-[4-(8-chloro-6-fluoro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000060_0001
20 Step 1: Preparation of l-(3-chloro-5-fluoro-2-hydroxy-phenyl)ethanone
Figure imgf000060_0002
20a
Compound 20a was prepared in analogy to the procedure described for the preparation of compound 15b by using 2-chloro-4-fluoro-phenol as the starting materials instead of 2-chloro-3- fluoro -phenol in Step 1.
Step 2: Preparation of 3-[2-[4-[(E)-3-(3-chloro-5-fluoro-2-hydroxy-phenyl)-3-oxo-prop-l- enyl] phenoxy ] ethoxy] cyclobutanecarboxylic acid
Figure imgf000061_0001
20b
Compound 20b was prepared in analogy to the procedure described for the preparation of compound 19b by using l-(3-chloro-5-fluoro-2-hydroxy-phenyl)ethanone as the starting materials instead of l-(3-chloro-2,4-dihydroxy-phenyl)ethanone in Step 2.
Step 3: Preparation of 3-[2-[4-(8-chloro-6-fluoro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000061_0002
20 To a solution of 3-[2-[4-[(£)-3-(3-chloro-5-fluoro-2-hydroxy-phenyl)-3-oxo-prop-l- enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid (100 mg, 0.23 mmol) in DMSO (2 mL) was added I2 (3 mg, O.Olmmol) at room temperature and then the mixture was stirred at 140 °C for 3 hours. After the reaction was completed, the reaction was cooled to room temperature, quenched with saturated NaHSCh solution (20 mL) and then extracted with EtOAc (20 mL) three times. The combined organic layer was washed with brine, dried over Na2S04 and concentrated in vacuo. The residue was then purified by preparative HPLC to give 3-[2-[4-(8-chloro-6-fluoro-4- oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid (500 mg , 20.1%) as a yellow solid.
Example 20:
Figure imgf000061_0003
NMR DMSO-d , 400MHz): d ppm 12.18 (s, 1H), 8.11-8.09 (m, 3H), 7.72-7.70 (m, 1H), 7.19-7.17 (m, 2H), 7.09-7.08 (m, 1H), 4.19-3.98 (m, 2H), 3.97-3.93 (m, 1H), 3.68-3.66 (m, 2H), 2.61-2.57 (m, 1H), 2.46-2.42 (m, 2H), 2.03-1.98 (m, 2H). MS obsd. (ESL) [(M+H)+]: 433.1. Example 21
3-[2-[4-(7,8-dichloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000062_0001
21 Example 21 was prepared in analogy to the procedure described for the preparation of
Example 15 by using 2,3-dichlorophenol as the starting materials instead of 2-chloro-3-fluoro- phenol in Step 1.
Example 21: lH NMR (DMSC )-d6, 400MHz): d ppm 12.17 (s, 1H), 8.08-8.00 (d, J= 3.6 Hz, 2H), 7.99-7.97 (d, 7=8.0 Hz, 1H), 7.75-7.73 (d, 7=8.0 Hz, 1H), 7.19-7.17 (d, 7=8.0 Hz, 2H), 7.1 (s, 1H), 4.21-4.19 (m, 2H), 3.68-3.66 (m, 3H), 2.59-2.50 (m, 1H), 2.46-2.45 (m, 2H), 2.43-2.40
(m, 1H), 2.01-1.98 (m, 1H). MS obsd. (ESI+) [(M+H)+]: 449.0.
Example 22
3-[2-[4-(8-chloro-6-cyano-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000062_0002
22
Step 1: Preparation of l-(5-bromo-3-chloro-2-hydroxy-phenyl)ethanone
Figure imgf000063_0001
22a
Compound 22a was prepared in analogy to the procedure described for the preparation of compound 15b by using 4-bromo-2-chloro-phenol as the starting materials instead of 2-chloro-3- fluoro -phenol in Step 1.
Step 2: Preparation of 6-bromo-8-chloro-2-(4-methoxyphenyl)chromen-4-one
Figure imgf000063_0002
Compound 22b was prepared in analogy to the procedure described for the preparation of compound 15d by using l-(5-bromo-3-chloro-2-hydroxy-phenyl)ethanone as the starting materials instead of l-(3-chloro-4-fluoro-2-hydroxy-phenyl)ethanone in Step 3.
Step 3: Preparation of 8-chloro-2-(4-methoxyphenyl)-4-oxo-chromene-6-carbonitrile
Figure imgf000063_0003
To a solution of 6-bromo-8-chloro-2-(4-methoxyphenyl)chromen-4-one (1.4 g, 2.74 mmol) in DMF (20 mL) was added Zn(CN)2 (643 mg, 5.47 mmol) and Pd(PPh3)4 (0.16 g, 0.140 mmol) under N2 atmosphere at room temperature. The mixture was stirred at 150 °C for 4 hours. After the reaction was completed, the mixture was poured into water (100 mL) and the resulting suspension was filtered. The solid was collected and washed with EtOH (10 mL) to give the crude 8-chloro-2-(4-methoxyphenyl)-4-oxo-chromene-6-carbonitrile (1.2 g, 100%) as a solid, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 312.0.
Step 4: Preparation of 8-chloro-2-(4-hydroxyphenyl)-4-oxo-chromene-6-carbonitrile
Figure imgf000064_0001
To a solution of 8-chloro-2-(4-methoxyphenyl)-4-oxo-chromene-6-carbonitrile (1.2 g, 3.9 mmol) in dichloro methane (20 ml) was added BBn (3.86 g, 15.4 mmol) at room temperature and the mixture was then stirred at room temperature overnight. After the reaction was completed, the reaction was quenched by adding into saturated NaHCCh solution (250 mL) slowly. The resulting suspension was filtered, the solid was collected and dried in vacuo to give the crude 8- chloro-2-(4-hydroxyphenyl)-4-oxo-chromene-6-carbonitrile (900 mg, 78.5%) as a yellow solid, which was used in the next step directly without further purification. (ESI+) [(M+H)+]: 297.9. Step 5: Preparation of 3-[2-[4-(8-chloro-6-cyano-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000064_0002
22
Example 22 was prepared in analogy to the procedure described for the preparation of Example 15 by using 8-chloro-2-(4-hydroxyphenyl)-4-oxo-chromene-6-carbonitrile as the starting materials instead of 8-chloro-7-fluoro-2-(4-hydroxyphenyl)chromen-4-one in Step 5. 100 mg of 3-[2-[4-(8-chloro-6-cyano-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxy lie acid was further purified by supercritical fluid chromatography (SFC) to give two diastereomers with cis- and trans- configuration, one of which is characterized as Example 22- A (45 mg) and the other is Example 22-B (45 mg).
Example 22: lH NMR (DMSO -d6, 400MHz): d ppm 12.19 (br s, 1H), 8.58 (d, J= 2.0 Hz, 1H), 8.43 - 8.36 (m, 1H), 8.10 (d, J= 8.9 Hz, 2H), 7.24 - 7.11 (m, 2H), 7.13 - 7.11 (m, 1H), 4.30 -
4.12 (m, 3H), 3.76 - 3.61 (m, 2H), 3.01 - 2.83 (m, 1H), 2.46 - 2.35 (m, 2H), 2.23 - 2.11 (m, 1H), 2.07 - 1.92 (m, 1H). MS obsd. (ESI+) [(M+H)+]: 440.2.
Example 22-A: lH NMR (DMSO -d6, 400MHz): d ppm 12.17 (br s, 1H), 8.58 (d, 7=2.0 Hz, 1H), 8.39 (d, 7=2.0 Hz, 1H), 8.10 (d, 7=9.0 Hz, 2H), 7.23 - 7.13 (m, 3H), 4.26 - 4.12 (m, 3H), 3.74 - 3.64 (m, 2H), 2.99 - 2.87 (m, 1H), 2.40 (ddd, 7=3.7, 6.9, 13.1 Hz, 2H), 2.23 - 2.10 (m, 2H).
MS obsd. (ESI+) [(M+H)+]: 440.2.
Example 22-B: lH NMR (DMSO-76, 400MHz): d ppm 12.30 - 11.80 (m, 1H), 8.61 - 8.55 (m, 1H), 8.43 - 8.35 (m, 1H), 8.16 - 8.05 (m, 2H), 7.25 - 7.13 (m, 3H), 4.20 (br s, 2H), 3.95 (q, 7=7.3 Hz, 1H), 3.73 - 3.64 (m, 2H), 2.63 - 2.56 (m, 1H), 2.47 - 2.38 (m, 2H), 2.06 - 1.93 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 440.2.
Example 23
3-[2-[4-(8-chloro-7-cyano-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000065_0001
23 Step 1: Preparation of l-(4-bromo-3-chloro-2-hydroxy-phenyl)ethanone
Figure imgf000065_0002
23a Compound 23a was prepared in analogy to the procedure described for the preparation of compound 15b by using 4-bromo-2-chloro-phenol as the starting materials instead of 2-chloro-3- fluoro -phenol in Step 1.
Step 2: Preparation of 7-bromo-8-chloro-2-(4-methoxyphenyl)chromen-4-one
Figure imgf000066_0001
23b
Compound 23b was prepared in analogy to the procedure described for the preparation of compound 15d by using l-(4-bromo-3-chloro-2-hydroxy-phenyl)ethanone as the starting materials instead of l-(3-chloro-4-fluoro-2-hydroxy-phenyl)ethanone in Step 3.
Step 3: Preparation of 8-chloro-2-(4-methoxyphenyl)-4-oxo-chromene-7-carbonitrile
Figure imgf000066_0002
23c
To a solution of 7-bromo-8-chloro-2-(4-methoxyphenyl)chromen-4-one (1.4 g, 2.74 mmol) in DMF (20 mL) was added Zn(CN)2 (643 mg, 5.47 mmol) and Pd(PPh3)4 (0.16 g, 0.140 mmol) under N2 atmosphere at room temperature. The mixture was stirred at 150 °C for 4 hours. After the reaction was completed, the mixture was poured into water (100 mL) and the resulting suspension was filtered. The solid was collected and washed with EtOH (10 mL) to give the crude 8-chloro-2-(4-methoxyphenyl)-4-oxo-chromene-7-carbonitrile (1.2 g, 100%) as a solid, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]:
312.0.
Step 4: Preparation of 8-chloro-2-(4-hydroxyphenyl)-4-oxo-chromene-7-carbonitrile
Figure imgf000067_0001
To a solution of 8-chloro-2-(4-methoxyphenyl)-4-oxo-chromene-7-carbonitrile (1.2 g, 3.9 mmol) in dichloro methane (20 ml) was added BBn (3.86 g, 15.4 mmol) at room temperature and the mixture was then stirred at room temperature overnight. After the reaction was completed, the reaction was quenched by adding into saturated NaHCCh solution (250 mL) slowly. The resulting suspension was filtered, the solid was collected and dried in vacuo to give the crude 8- chloro-2-(4-hydroxyphenyl)-4-oxo-chromene-7-carbonitrile (900 mg, 78.5%) as a yellow solid, which was used in the next step directly without further purification. (ESI+) [(M+H)+]: 297.9. Step 5: Preparation of 3-[2-[4-(8-chloro-7-cyano-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000067_0002
23
Example 23 was prepared in analogy to the procedure described for the preparation of Example 15 by using 8-chloro-2-(4-hydroxyphenyl)-4-oxo-chromene-7-carbonitrile as the starting materials instead of 8-chloro-7-fluoro-2-(4-hydroxyphenyl)chromen-4-one in Step 5. 45 mg of 3-[2-[4-(8-chloro-7-cyano-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid was further purified by supercritical fluid chromatography (SFC) to give two diastereomers with cis- and trans- configuration, one of which is characterized as Example 23-A (31 mg) and the other is Example 23-B (11 mg).
Example 23:
Figure imgf000067_0003
NMR (DMSC )-d6, 400MHz): d ppm 12.19 (br s, 1H), 8.10 (dd, 7=2.7, 8.1 Hz, 3H), 8.00 (d, 7=8.2 Hz, 1H), 7.32 - 7.09 (m, 3H), 4.26 - 4.12 (m, 2H), 3.95 ( q , 7=7.3 Hz, 1H), 3.73 - 3.63 (m, 2H), 3.01 - 2.83 (m, 1H), 2.45 - 2.36 (m, 2H), 2.23 - 2.10 (m, 1H), 2.07 - 1.93 (m, 1H). MS obsd. (ESE) [(M+H)+]: 440.0.
Example 23-A:
Figure imgf000068_0001
NMR DMSO-d , 400MHz): d ppm 8.14 - 8.08 (m, 3H), 8.00 (d, 7=8.2 Hz, 1H), 7.22 - 7.16 (m, 3H), 4.22 - 4.18 (m, 2H), 3.94 (s, 1H), 3.73 - 3.64 (m, 2H), 2.57 (br s, 1H), 2.44 (br d, 7=9.2 Hz, 2H), 2.03 - 1.95 (m, 2H). MS obsd. (ESE) [(M+H)+]: 440.0.
Example 23-B: lH NMR (DMSO-76, 400MHz): d ppm 8.17 - 8.07 (m, 3H), 8.00 (d, 7=8.3 Hz, 1H), 7.24 - 7.15 (m, 3H), 4.27 - 4.12 (m, 3H), 3.72 - 3.64 (m, 2H), 3.00 - 2.86 (m, 1H), 2.39 (ddd, 7=3.7, 6.9, 13.1 Hz, 2H), 2.23 - 2.11 (m, 2H). MS obsd. (ESE) [(M+H)+]: 440.2.
Example 24
C7s-3-[2-[4-(8-chloro-5-hydroxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000068_0003
Step 1: Preparation of l-(3-chloro-2,6-dihydroxy-phenyl)ethanone
Figure imgf000068_0002
24a
To a solution of l-(2,6-dihydroxyphenyl)ethanone (5g, 32.9 mmol) in AcOH (40 mL) was added NCS (4.83 g, 36.1 mmol) at room temperature and then the mixture was stirred at 50°C for 2hours. After the reaction was completed, the mixture was concentrated in vacuo and the residue was purified by column chromatography on silica gel (elution with PE:EtOAc = 5: 1) to give l-(3-chloro-2,6-dihydroxy-phenyl)ethanone (200 mg 3.26 % yield) as a white solid. MS obsd. (ESI+) [(M+H)+] : 187.1.
Step 2: Preparation of l-[3-chloro-6-hydroxy-2-(2- methoxyethoxymethoxy)phenyl]ethanone
Figure imgf000069_0001
24b
A mixture of l-(3-chloro-2,6-dihydroxy-phenyl)ethanone (6 g, 32.2 mmol), 1- (chloromethoxy)-2-methoxyethane (4.81 g, 38.6 mmol) and K2C03(8.89 g, 3.88 ml, 64.3 mmol) in DMF (20 mL) was stirred at room temperature for 16 hours. After the reaction was completed, the mixture was diluted with water (50 mL) and the resulting mixture was extracted with dichloromethane (50 mL) three times. The combined organic layer was washed with brine, dried over Na2S04 and concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with PE:EtOAc = 10: 1 to 3:1) to give l-[3-chloro-6-hydroxy-2-(2- methoxyethoxymethoxy)phenyl]ethanone (8.0 g, 90.6 %) as a colorless oil. MS obsd. (ESI+) [(M+H)+]: 275.1.
Step 3: Preparation of ,s-3-[2-[4-[(/i)-3-[3-chloro-6-hydroxy-2-(2- methoxyethoxymethoxy)phenyl]-3-oxo-prop-l-enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000069_0002
A mixture of l-[3-chloro-6-hydroxy-2-(2-methoxyethoxymethoxy)phenyl]ethanone (1.3 g, 4.73 mmol), r/.v-ethyl 3-(2-(4-formylphenoxy)ethoxy)cyclobutanecarboxylate (int-4, 1.38 g, 4.73 mmol) and KOH (1.06 g, 18.9 mmol) in EtOH (40 mL) was stirred at room temperature for 16 hours. After the reaction was completed, the mixture was quenched with ice-water (60 mL) and adjusted to pH~6 by addition of 2N HC1. The resulting mixture was extracted with EtOAc (100 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo to give the crude cA-3-[2-[4-[(£’)-3-[3-chloro-6-hydroxy-2-(2- methoxyethoxymethoxy)phenyl]-3-oxo-prop-l-enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid (2.6 g, 100 %) as a yellow solid, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+] : 521.1.
Step 4: Preparation of .s-3-[2-[4-(8-chloro-5-hydroxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000070_0001
24
To a solution of cA-3-[2-[4-[(£)-3-[3-chloro-6-hydroxy-2-(2- methoxyethoxymethoxy)phenyl]-3-oxo-prop-l-enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid (100 mg, l92umol) in DMSO (60 mL) was added I2 (5 mg, 19.2 pmol) and the mixture was stirred at 140 °C for 3 hours. After the reaction was completed, the reaction was cooled to room temperature, quenched with saturated NaHSCL solution (10 mL) and diluted with water 100 mL. The resulting suspension was filtered, the solid was collected and purified by preparative HPLC to afford cA-3-[2-[4-(8-chloro-5-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (20mg, 24.2 % yield) as a yellow solid. 1 H NMR (DMSO-^6, 400MHZ): d ppm 12.88 - 12.62 (br s, 1H), 12.32 - 11.91 (br s, 1H), 8.10 (d, J= 9.0 Hz, 2H), 7.81 (d, J= 8.9 Hz, 1H), 7.22 - 7.10 (m, 3H), 6.84 (d, 7=8.9 Hz, 1H), 4.25 - 4.13 (m, 2H),
3.95 (q, J=7.3 Hz, 1H), 3.70 - 3.64 (m, 2H), 2.64 - 2.56 (m, 1H), 2.47 - 2.37 (m, 2H), 2.06 - 1.95 (m, 2H). MS obsd. (ESL) [(M+H)+]: 431.1.
Example 25
C¾-3-[2-[4-(8-chloro-5-methoxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000071_0001
25
Example 25 was prepared in analogy to the procedure described for the preparation of Example 19 by using c/5'-3-[2-[4-(8-chloro-5-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid as the starting materials instead of 3-[2-[4-(8- chloro-7-hydroxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
(intermediate 19c) in Step 1.
Example 25: lH NMR (DMSC )-d6, 400MHz): d ppm 12.73 (br s, 1H), 8.02 (d, 7=8.93 Hz, 2H), 7.85 (d, 7=9.05 Hz, 1H), 7.14 (d, 7=8.93 Hz, 2H), 7.01 (d, 7=9.05 Hz, 1H), 6.86 (s, 1H), 4.13-4.25 (m, 2H), 3.91-3.99 (m, 1H), 3.87 (s, 3H), 3.63-3.72 (m, 2H), 2.56-2.64 (m, 1H), 2.36-
2.48 (m, 2H), 1.93-2.05 (m, 2H). MS obsd. (ESI+) [(M+H)+]:445.2.
Example 27
Ci,s-3-[2-[4-(8-chloro-5-isopropoxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000071_0002
27 Example 27 was prepared in analogy to the procedure described for the preparation of Example 19 by using c/5'-3-[2-[4-(8-chloro-5-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid and 2-iodopropane as the starting materials instead of 3-[2-[4-(8-chloro-7-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (intermediate 19c) and iodomethane in Step 1.
Example 27: lH NMR (DMSC )-d6, 400MHz): d ppm 12.17 (br s, 1H), 8.03 (d, 7=9.05 Hz, 2H), 7.81 (d, 7=9.05 Hz, 1H), 7.15 (d, 7=8.93 Hz, 2H), 7.02 (d, 7=9.17 Hz, 1H), 6.82 (s, 1H), 4.61-4.74 (m, 1H), 4.12-4.23 (m, 2H), 3.93 ( q , 7=7.37 Hz, 1H), 3.63-3.69 (m, 2H), 2.53-2.61 (m, 1H), 2.36-2.47 (m, 2H), 1.93-2.04 (m, 2H), 1.32 (d, 7=5.99 Hz, 6H). MS obsd. (ESI+)
[(M+H)+]:473.3.
Example 28
C¾-3-[2-[4-(5-bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000072_0001
28
Step 1: Preparation of l-(6-bromo-3-chloro-2-hydroxy-phenyl)ethanone
Figure imgf000072_0002
28a
Compound 28a was prepared in analogy to the procedure described for the preparation of compound 15b by using 5-bromo-2-chloro-phenol as the starting materials instead of 2-chloro-3- fluoro -phenol in Step 1. Step 2: Preparation of .s-3-[2-[4-[(/i)-3-(6-bronio-3-chloro-2-hydroxy-phenyl)-3-oxo-prop- 1 -enyl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000073_0001
28b A mixture of l-(6-bromo-3-chloro-2-hydroxy-phenyl)ethanone (230 mg, 0.9 mmol), cis- ethyl 3-(2-(4-formylphenoxy)ethoxy)cyclobutanecarboxylate (268 mg, 0.9 mmol) and KOH (207 mg, 3.7 mmol) in EtOH (40 mL) was stirred at room temperature for 16 hours. After the reaction was completed, the mixture was quenched with ice-water (60 mL) and adjusted to pH~6 by addition of 2N HC1. The resulting mixture was extracted with EtOAc (100 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo to give the crude cA-3-[2-[4-[(£)-3-(6-bromo-3-chloro-2-hydroxy-phenyl)-3-oxo-prop- l-enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid (300 mg, 65.6 %) as a yellow solid, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 495.2. Step 3: Preparation of Cis-3-[2-[4-(5-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000073_0002
28
Example 28 was prepared in analogy to the procedure described for the preparation of Example 19 by using cis-3-[2-[4-[(£’)-3-(6-bromo-3-chloro-2-hydroxy-phenyl)-3-oxo-prop-l- enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid as the starting materials instead of 3-[2-[4- [(£)-3-(3-chloro-2, 4-dihydro xy-phenyl)-3-oxo-prop-l- enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid (intermediate 19b) in Step 3.
Example 28:
Figure imgf000074_0001
NMR DMSO-d , 400MHz): d ppm 12.15 (s, 1H), 8.00-8.15 (m, 2H), 7.85 (d, 7=8.56 Hz, 1H), 7.69 (d, 7=8.56 Hz, 1H), 7.16 (d, 7=9.05 Hz, 2H), 7.05 (s, 1H), 4.13- 4.28 (m, 2H), 3.87-4.04 (m, 1H), 3.62-3.71 (m, 2H), 2.54-2.64 (m, 1H), 2.39-2.48 (m, 2H), 1.94-
2.04 (m, 2H). MS obsd. (ESI+) [(M+H)+]:493.2.
Example 29
C¾-3-[2-[4-(5,8-dichloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000074_0002
29
Step 1: Preparation of l-(3,6-dichloro-2-hydroxy-phenyl)ethanone
Figure imgf000074_0003
29a
Compound 29a was prepared in analogy to the procedure described for the preparation of compound 15b by using 2,5-dichlorophenol as the starting materials instead of 2-chloro-3- fluoro -phenol in Step 1.
Step 2: Preparation of C¾-3-[2-[4-(5,8-dichloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000075_0001
29
Example 29 was prepared in analogy to the procedure described for the preparation of Example 19 by using l-(3,6-dichloro-2-hydroxy-phenyl)ethanone and r/.v-ethyl 3-(2-(4- formylphenoxy)ethoxy)cyclobutanecarboxylate as the starting materials instead of l-(3-chloro- 2, 4-dihydro xy-phenyl)ethanone (intermediate 19a) and methyl 3 -[2- (4- formylphenoxy)ethoxy]cyclobutanecarboxylate in Step 2.
Example 29: lH NMR (DMSC )-d6, 400MHz): d ppm 12.17 (br s, 1H), 7.98-8.06 (m, 2H), 7.89 (d, 7=8.56 Hz, 1H), 7.46 (d, 7=8.56 Hz, 1H), 7.12 (d, 7=9.05 Hz, 2H), 6.97 (s, 1H), 4.12- 4.23 (m, 2H), 3.95 ( q , 7=7.34 Hz, 1H), 3.62-3.73 (m, 2H), 2.54-2.66 (m, 1H), 2.41-2.49 (m, 2H),
1.94-2.06 (m, 2H). MS obsd. (ESE) [(M+H)+]:447.2.
Example 30
Ci.s-3-[2-[4-(8-chloro-5-cyclopropyl-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000075_0002
30
A mixture of cA-3-[2-[4-(5-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (50mg, 101 m mol), cyclopropylboronic acid (26.1 mg, 304 pmol), K3PO4 (43 mg, 203 pmol) and Pd(dppf)Cl2 (37 mg, 50.6 pmol) in DMF (5 mL) was heated at 100 °C under microwave radiation for 1 hour. After the reaction was completed, the mixture was concentrated in vacuo and the residue was purified by preparative HPLC to give c/5'-3-[2-[4-(8-chloro-5-cyclopropyl-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (14 mg, 30.4%) as a white solid. 1 H NMR
(DMSO-rfc, 400MHz): d ppm 12.17 (br s, 1H), 8.07 (d, 7=8.93 Hz, 2H), 7.79 (d, 7=8.44 Hz, 1H), 7.17 (d, 7=9.05 Hz, 2H), 6.90-7.01 (m, 2H), 4.13-4.25 (m, 2H), 3.93 (q, 7=7.37 Hz, 1H), 3.64-
3.70 (m, 2H), 3.55-3.64 (m, 1H), 2.54-2.61 (m, 1H), 2.38-2.47 (m, 2H), 1.94-2.04 (m, 2H), 1.01- 1.08 (m, 2H), 0.74-0.81 (m, 2H). MS obsd. (ESI+) [(M+H)+]:445. l.
Example 31
C¾-3-[2-[4-(8-chloro-5-methyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000076_0001
31
Example 31 was prepared in analogy to the procedure described for the preparation of Example 30 by using 2,4,6-trimethyl- 1,3, 5, 2, 4, 6-trio xatriborinane as the starting materials instead of cyclopropylboronic acid.
Example 31:
Figure imgf000076_0002
NMR DMSO-d , 400MHz): d ppm 8.05 (d, 7=9.0 Hz, 2H), 7.81 (d,
7=8.1 Hz, 1H), 7.22 (dd, 7=0.7, 8.1 Hz, 1H), 7.15 (d, 7=9.0 Hz, 2H), 6.95 (s, 1H), 4.21 - 4.15 (m, 2H), 3.95 (s, 1H), 3.69 - 3.64 (m, 2H), 2.74 (s, 3H), 2.58 (d, 7=8.1 Hz, 1H), 2.48 - 2.39 (m, 2H), 2.04 - 1.94 (m, 2H). MS obsd. (ESE) [(M+H)+]:429.2.
Example 33
C¾-3-[2-[4-(8-bromo-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000077_0001
33
Example 33 was prepared in analogy to the procedure described for the preparation of Example 19 by using l-(3-bromo-2-hydroxy-phenyl)ethanone and r/.v-ethyl 3-(2-(4- formylphenoxy)ethoxy)cyclobutanecarboxylate as the starting materials instead of l-(3-chloro- 2, 4-dihydro xy-phenyl)ethanone (intermediate 19a) and methyl 3 -[2- (4- formylphenoxy)ethoxy]cyclobutanecarboxylate in Step 2.
Example 33: lH NMR (DMSC )-d6, 400MHz): d ppm 12.20 (br s, 1H), 8.07-8.18 (m, 3H), 8.03 (dd, 7=1.53, 7.89 Hz, 1H), 7.43 (t, 7=7.82 Hz, 1H), 7.14-7.21 (m, 2H), 7.03-7.10 (m, 1H), 4.16-4.25 (m, 2H), 3.89-4.01 (m, 1H), 3.62-3.71 (m, 2H), 2.55-2.64 (m, 1H), 2.40-2.48 (m, 2H),
1.94-2.05 (m, 2H). MS obsd. (ESE) [(M+H)+]:459.3.
Example 34
C¾-3-[2-[4-(8-cyclopropyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000077_0002
34
Example 34 was prepared in analogy to the procedure described for the preparation of Example 30 by using 3-[2-[4-(8-bromo-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid as the starting materials instead of cis-3-[2-[4-(5- bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid. Example 34:
Figure imgf000078_0001
NMR (DMSO -d6, 400MHz): d ppm 11.52-13.15 (m, 1H), 8.10 (d, 7=8.93 Hz, 2H), 7.84 (dd, 7=1.83, 7.58 Hz, 1H), 7.33-7.45 (m, 2H), 7.15 (d, 7=8.93 Hz, 2H), 6.98 (s, 1H), 4.13-4.25 (m, 2H), 3.94 ( q , 7=7.34 Hz, 1H), 3.60-3.73 (m, 2H), 2.52-2.64 (m, 2H), 2.40- 2.48 (m, 2H), 1.93-2.06 (m, 2H), 1.07-1.26 (m, 2H), 0.74-0.96 (m, 2H). MS obsd. (ESI+)
[(M+H)+]:42l.2.
Example 35
C¾-3-[2-[4-(8-methyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000078_0002
35 Example 35 was prepared in analogy to the procedure described for the preparation of
Example 30 by using 3-[2-[4-(8-bromo-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid and 2,4,6-trimethyl- 1,3, 5, 2, 4, 6-trio xatriborinane as the starting materials instead of cis-3-[2-[4-(5-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid and 2,4,6-trimethyl- 1,3, 5, 2, 4, 6-trio xatriborinane.
Example 35: lH NMR (DMSO-76, 400MHz): d ppm 12.16 (br s, 1H), 8.07 (d, 7=8.93 Hz,
2H), 7.83-7.93 (m, 1H), 7.68 (d, 7=7.21 Hz, 1H), 7.38 (t, 7=7.58 Hz, 1H), 7.15 (d, 7=9.05 Hz, 2H), 6.96 (s, 1H), 4.13-4.26 (m, 2H), 3.95 (q, 7=7.37 Hz, 1H), 3.61-3.74 (m, 2H), 2.55-2.66 (m, 4H), 2.39-2.49 (m, 2H), 1.93-2.05 (m, 2H). MS obsd. (ESI+) [(M+H)+]:395.2.
Example 37 2-[2-[4-(8-chloro-5-fluoro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetic acid
Figure imgf000078_0003
37
Step 1: Preparation of l-(3-chloro-6-fluoro-2-hydroxy-phenyl)ethanone
Figure imgf000079_0001
37a Compound 37a was prepared in analogy to the procedure described for the preparation of compound 15b by using 2-chloro-5-fluoro-phenol as the starting materials instead of 2-chloro-3- fluoro -phenol in Step 1.
Step 2: Preparation of 8-chloro-5-fluoro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one
Figure imgf000079_0002
Compound 37b was prepared in analogy to the procedure described for the preparation of compound la by using l-(3-chloro-6-fluoro-2-hydroxy-phenyl)ethanone as the starting material instead of l-(3-chloro-2-hydroxy-phenyl)ethanone in Step 1.
Step 3: Preparation of 2-[2-[4-(8-chloro-5-fluoro-4-oxo-chromen-2- yl)phenoxy] ethoxy] acetic acid
Figure imgf000079_0003
37 Example 37 was prepared in analogy to the procedure described for the preparation of Example 9 by using 8-chloro-5-fluoro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one as the starting material instead of 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one in Step 1.
Example 37:
Figure imgf000080_0001
11.92 (m, 1H), 8.07 (d, 7=9.0 Hz, 2H), 8.00 (dd, 7=4.9, 8.8 Hz, 1H), 7.30 (dd, 7=8.9, 10.6 Hz, 1H), 7.18 (d, 7=9.0 Hz, 2H), 7.01 (s, 1H), 4.27 - 4.20 (m, 2H), 4.09 (s, 2H), 3.90 - 3.81 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 393.1.
Example 38-A and Example 38-B
C¾-3-[2-[[5-(8-chloro-4-oxo-chromen-2-yl)-2-pyridyl]oxy]ethoxy]cyclobutanecarboxylic acid and h' «.s-3-[2-[[5-(8-chloro-4-oxo-chromen-2-yl)-2- pyridy 1] oxy ] ethoxy] cy clobutanecarboxy lie acid
Figure imgf000080_0002
38-A and 38-B
Step 1: Preparation of (2-acetyl-6-chloro-phenyl) 6-chloropyridine-3-carboxylate
Figure imgf000080_0003
38a
To a solution of l-(3-chloro-2-hydroxy-phenyl)ethanone (5g, 29.3 mmol) and TEA(3.56 g, 4.91 ml, 35.2 mmol) in dichloromethane (20 mL) was added 6-chloropyridine-3-carbonyl chloride (5.42 g, 30.8 mmol). The reaction was stirred at room temperature for 14 hours. After the reaction was completed, the solution was concentrated in vacuo to give the crude (2-acetyl-6- chloro -phenyl) 6-chloropyridine-3-carboxylate (9.1 g, 100%) as a yellow oil, which was used in the next step directly without further ourification. MS obsd. (ESI+) [(M+H)+]: 310.1.
Step 2: Preparation of l-(3-chloro-2-hydroxy-phenyl)-3-(6-chloro-3-pyridyl)propane-l,3- dione
Figure imgf000081_0001
38b
To a solution of (2-acetyl-6-chloro-phenyl) 6-chloropyridine-3-carboxylate (9g, 29 mmol) in THF (400 ml) was added potassium te/t-butoxide (4.56 g, 40.6 mmol) at room temperature. Then mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction was quenched with water (50 mL) and the aqueous was adjusted to pH~6 by addition of 0.5N HC1. The resulting mixture was then extracted with dichloromethane (100 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo. The residue was then triturated in hexane (15 mL) and the mixture was then filtred. The solid was collected and dried in vacuo to give l-(3-chloro-2-hydroxy-phenyl)-3- (6-chloro-3-pyridyl)propane-l,3-dione (9.0 g, 100%) as a light brown solid, which was used in the next step directly without further ourification. MS obsd. (ESI+) [(M+H)+]: 310.1.
Step 3: Preparation of 8-chloro-2-(6-chloro-3-pyridyl)chromen-4-one
Figure imgf000081_0002
38c
To a mixture of l-(3-chloro-2-hydroxy-phenyl)-3-(6-chloro-3-pyridyl)propane-l,3-dione (2.5 g, 8.06 mmol) in AcOH (15 mL) was added cat. con.LLSCL (2 drop). The mixture was then stirred l30°C for 4 hours. After the reaction was completed, the mixture was concentrated in vacuo to remove the solvent and the residue was suspended in water (20 mL). The suspension was then filtered, the solid was collected and dried in vacuo to give 8-chloro-2-(6-chloro-3- pyridyl)chromen-4-one (2.0 g, 84.9%) as a white solid, which was used in the next step directly without further ourification. MS obsd. (ESI+) [(M+H)+]: 292.1.
Step 4: Preparation of ,s-3-[2-[[5-(8-chloro-4-oxo-chromen-2-yl)-2-pyridyl]oxy]ethoxy]cyclobutanecarboxylic acid and frans-3-[2-[[5-(8-chloro-4-oxo-chromen-2-yl)-2- pyridy 1] oxy ] ethoxy] cy clobutanecarboxy lie acid
Figure imgf000082_0001
38-A and 38-B
To a solution of methyl 3-(2-hydroxyethoxy)cyclobutanecarboxylate (1.91 g,l6 mmol) in DMF (20 mL) was added NaH (986 mg, 24.6 mmol) and the mixture was stirred at room temperature for 0.5 hour.Then the resulting mixture was added into the solution of 8-chloro-2-(6- chloro-3-pyridyl)chromen-4-one (1.6 g, 5.48 mmol) in DMF (20mL). After addition, the mixture was stirred at 80°C for 3 hours. After the reaction was completed, the mixture was filtred and the filtrate was purified by preparative HPLC to give 3-[2-[[5-(8-chloro-4-oxo-chromen-2-yl)-2- pyridyl] oxy] ethoxy] cyclobutanecarboxy lie acid (320 mg, 14.0%) as a white solid. The solid was then further purified by supercritical fluid chromatography (SFC) to give two diastereomers with cis- and Irons- configuration, one of which is characterized as Example 38-A (175 mg, 7.68%) and the other is Example 38-B (40 mg, 1.76%).
Example 38-A: lH NMR (DMSO -d6, 400MHz): d ppml2.62 (s, 1H), 8.85-8.95 (m, 1H), 8.30-8.38 (m, 1H), 8.00-8.07 (m, 1H), 7.89-7.96 (m, 1H), 7.43-7.51 (m, 1H), 6.97-7.06 (m, 2H), 4.44-4.54 (m, 2H), 3.92-4.04 (m, 1H), 3.63-3.78 (m, 2H), 2.59-2.67 (m, 1H), 2.43-2.53 (m, 2H), 1.98-2.17 (m, 2H). MS obsd. (ESF) [(M+H)+]: 416.1. Example
Figure imgf000083_0001
ppm 11.72-12.61 (m, 1H), 8.87-8.96 (m, 1H), 8.33-8.43 (m, 1H), 7.99 (dd, 7=1.59, 7.83 Hz, 2H), 7.49 (s, 1H), 7.15 (s, 1H), 7.03-7.10 (m, 1H), 4.42-4.53 (m, 2H), 4.09-4.24 (m, 1H), 3.63-3.71 (m, 2H), 2.87-2.98 (m, 1H), 2.34-2.44 (m, 2H), 2.08-2.23 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 416.1.
Example 39 and Example 40
C¾-3-[2-[5-(8-chloro-4-oxo-chromen-2-yl)pyrazin-2-yl]oxyethoxy]cyclobutanecarboxylic acid and Cis-methyl 3-[2-[5-(8-chloro-4-oxo-chromen-2-yl)pyrazin-2- yl] oxy ethoxy ] cyclobutanecarboxy late
Figure imgf000083_0002
40
Step 1: Preparation of 5-chloropyrazine-2-carbonyl chloride
Figure imgf000083_0003
To a solution of 5-chloropyrazine-2-carboxylic acid (3 g, 18.9 mmol) in dichloromethane (60 mL) was added oxalyl dichloride (2.52 g, 1.7 mL, 19.9 mmol) dropwise at 0°C. Then to the mixture was added 2 drops of DMF and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the mixture was concentrated in vacuo to give 5- chloropyrazine-2-carbonyl chloride (3.4 g, 100%) as a white solid.
Step 2: Preparation of 8-chloro-2-(5-chloropyrazin-2-yl)chromen-4-one
Figure imgf000084_0001
Compound 39b was prepared in analogy to the procedure described for the preparation of compound 38c by using 5-chloropyrazine-2-carbonyl chloride as the starting material instead of 6-chloropyridine-3-carbonyl chloride chloride in Step 1.
Step 3: Preparation of ci, v-3-[2-[5-(8-chloro-4-oxo-chromen-2-yl)pyrazin-2- yl] oxy ethoxy ]cyclobutanecarboxy lie acid and civ- methyl 3-[2-[5-(8-chloro-4-oxo-chromen- 2-yl)pyrazin-2-yl]oxyethoxy]cyclobutanecarboxylate
Figure imgf000084_0002
40 To a mixture of methyl 3-(2-hydroxyethoxy)cyclobutanecarboxylate (166 mg, 955 mihoΐ) in DMF (5 mL) was added NaH (34.5 mg, 863 mihoΐ) and the mixture was stirred at room temperature for 0.5 hour.Then to the resulting mixture was added the solution of 8-chloro-2-(5- chloropyrazin-2-yl)chromen-4-one (140 mg, 479 m mol) in DMF (5 mL). After addition, the mixture was stirred at room temperature for 12 hours. After the reaction was completed, the mixture was filtred and the filtrate was purified by preparative HPLC to give the cis
configuration of 3-[2-[5-(8-chloro-4-oxo-chromen-2-yl)pyrazin-2- yl]oxyethoxy]cyclobutanecarboxylic acid (5 mg, 2.5%) and methyl 3-[2-[5-(8-chloro-4-oxo- chromen-2-yl)pyrazin-2-yl]oxyethoxy]cyclobutanecarboxylate (14 mg, 6.8%) as white foam. The trans isomer was not collected in the purification.
Example 39:
Figure imgf000085_0001
-8.98 (m, 1H), 8.52-8.62 (m, 1H),
7.99-8.09 (m, 2H), 7.43-7.58 (m, 1H), 7.02-7.13 (m, 1H), 4.50-4.56 (m, 2H), 3.92-3.99 (m, 1H), 3.68-3.75 (m, 2H), 2.55-2.63 (m, 1H), 2.39-2.48 (m, 2H), 1.94-2.05 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 417.1.
Example 40:
Figure imgf000085_0002
8.59
(m, 1H), 8.00-8.07 (m, 2H), 7.49-7.56 (m, 1H), 7.07-7.11 (m, 1H), 4.50-4.55 (m, 2H), 3.93-4.02 (m, 1H), 3.68-3.76 (m, 2H), 3.63 (s, 3H), 2.67-2.73 (m, 1H), 2.42-2.48 (m, 2H), 1.98-2.05 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 431.1.
Example 41 Civ-methyl 3-[2-[6-(8-chloro-4-oxo-chromen-2-yl)pyridazin-3- yl] oxy ethoxy ] cyclobutanecarboxy late
Figure imgf000085_0003
41
Step 1: Preparation of 6-chloropyridazine-3-carbonyl chloride
Figure imgf000086_0001
41a
To a solution of 6-chloropyridazine-3-carboxylic acid (4.4 g, 27,8 mmol) in
dichloromethane (60 mL) was added oxalyl dichloride (3.7 g, 2.5 mL, 29.1 mmol) dropwise at 0°C. Then to the mixture was added 2 drops of DMF and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the mixture was concentrated in vacuo to give 5-chloropyrazine-2-carbonyl chloride (5.0g, 100%) as a yellow solid.
Step 2: Preparation of 8-chloro-2-(6-chloropyridazin-3-yl)chromen-4-one
Figure imgf000086_0002
41b
Compound 41b was prepared in analogy to the procedure described for the preparation of compound 38c by using 6-chloropyridazine-3-carbonyl chloride as the starting material instead of 6-chloropyridine-3-carbonyl chloride chloride in Step 1.
Step 3: Preparation of .v- methyl 3-[2-[6-(8-chloro-4-oxo-chromen-2-yl)pyridazin-3- yl] oxy ethoxy ] cyclobutanecarboxy late
Figure imgf000086_0003
41 To a mixture of methyl 3-(2-hydroxyethoxy)cyclobutanecarboxylate (166 mg, 955 mihoΐ) in DMF (5 mL) was added NaH (34.5 mg, 863 mmol) and the mixture was stirred at room temperature for 0.5 hour. Then to the resulting mixture was added the solution of 8-chloro-2-(6- chloropyridazin-3-yl)chromen-4-one (140 mg, 479 m mol) in DMF (5 mL). After addition, the mixture was stirred at room temperature for 12 hours. After the reaction was completed, the mixture was filtred and the filtrate was purified by preparative HPLC to give the cis
configuration of methyl 3-[2-[5-(8-chloro-4-oxo-chromen-2-yl)pyrazin-2- yl]oxyethoxy]cyclobutanecarboxylate (60 mg, 29.2%) as white foam, the trans isomer was not collected in the purification.
Example 41:
Figure imgf000087_0001
8.08
(m, 2H), 7.51-7.61 (m, 2H), 7.32 (s, 1H), 4.63-4.69 (m, 2H), 3.94-4.02 (m, 1H), 3.71-3.77 (m, 2H), 3.60 (s, 3H), 2.65-2.74 (m, 1H), 2.41-2.48 (m, 2H), 1.94-2.09 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 431.1.
Example 42 Ci,s-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-fluoro-phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000087_0002
42
Step 1: Preparation of 8-chloro-2-(3-fluoro-4-methoxy-phenyl)chromen-4-one
Figure imgf000087_0003
42a Compound 42a was prepared in analogy to the procedure described for the preparation of compound 38c by using 3-fluoro-4-methoxy-benzoyl chloride as the starting material instead of 6-chloropyridine-3-carbonyl chloride chloride in Step 1.
Step 2: Preparation of 8-chloro-2-(3-fluoro-4-hydroxy-phenyl)chromen-4-one
Figure imgf000088_0001
42b
To a solution of 8-chloro-2-(3-fluoro-4-methoxy-phenyl)chromen-4-one (850 mg, 2.7 mmol) in dichloro methane (20 mL) was added B B n (1.0 M in dichloromethane, 13 mL, 13 mmol) at room temperature and the mixture was then stirred at room temperature overnight.
After the reaction was completed, the reaction was quenched by adding into saturated NaHCCh solution (250 mL) slowly. The resulting suspension was filtered, the solid was collected and dried in vacuo to give the crude 8-chloro-2-(3-fluoro-4-hydroxy-phenyl)chromen-4-one (250 mg) as a yellow solid, which was used in the next step directly without further purification. (ESI+) [(M+H)+]: 291.1.
Step 3: Preparation of .s-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-fluoro- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000088_0002
42
To a mixture of 8-chloro-2-(3-fluoro-4-hydroxy-phenyl)chromen-4-one (180 mg, 619 m mol) and methyl 3-(2-(tosyloxy)ethoxy)cyclobutanecarboxylate (224 mg, 681 m mol,) in DMF (5 mL) was added K2CO3 (171 mg, 1.24 mmol) and the mixture was then stirred at 50°C overnight. Then to the resulting mixture was added MeOH (5 mL), water (1 mL) and LiOH (44.5 mg, 1.86 mmol). After the addition, the mixture was stirred at room temperature for 4 hours. The mixture was then filtered and the filtrate was purified by preparative HPLC to give the cis configuration of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-fluoro- phenoxy] ethoxy] cyclobutanecarboxy lie acid (15 mg, 5.6%) as a white foam, the trans isomer was not collected in the purification. 1 H NMR (DMSO-J^, 400MHz): d ppm 7.90-8.04 (m, 3H), 7.34-7.55 (m, 3H), 7.14 (s, 1H), 4.23-4.32 (m, 2H), 3.90-4.02 (m, 1H), 3.64-3.75 (m, 2H), 2.55- 2.65 (m, 1H), 2.39-2.47 (m, 2H), 1.92-2.06 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 433.1.
Example 43 C¾-3-[2-[2-chloro-4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000089_0001
43
Step 1: Preparation of 8-chloro-2-(3-chloro-4-methoxy-phenyl)chromen-4-one
Figure imgf000089_0002
43a
Compound 43a was prepared in analogy to the procedure described for the preparation of compound 38c by using 3-chloro-4-methoxy-benzoyl chloride as the starting material instead of 6-chloropyridine-3-carbonyl chloride chloride in Step 1.
Step 2: Preparation of '.s-3-[2-[2-chloro-4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000090_0001
43
Example 43 was prepared in analogy to the procedure described for the preparation of Example 42 by using 8-chloro-2-(3-chloro-4-methoxy-phenyl)chromen-4-one as the starting material instead of 8-chloro-2-(3-fluoro-4-methoxy-phenyl)chromen-4-one in Step 2.
Example 43: lH NMR (DMSO -d6, 400MHz): d ppm 12.02-12.44 (m, 1H), 8.15-8.21 (m, 1H), 8.05-8.10 (m, 1H), 7.96-8.02 (m, 2H), 7.45-7.54 (m, 1H), 7.34-7.40 (m, 1H), 7.14-7.21 (m, 1H), 4.26-4.31 (m, 2H), 3.92-4.07 (m, 1H), 3.67-3.77 (m, 2H), 2.55-2.63 (m, 1H), 2.39-2.47 (m, 2H), 1.96-2.06 (m, 2H). MS obsd. (ESE) [(M+H)+]: 449.1.
Example 44
C¾-3-[2-[2-bromo-4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000090_0002
44 Step 1: Preparation of 2-(3-bromo-4-hydroxy-phenyl)-8-chloro-chromen-4-one
Figure imgf000090_0003
44a
Compound 44a was prepared in analogy to the procedure described for the preparation of compound 3c by using 3-bromo-4-methoxy-benzaldehyde as the starting material instead of 4- methoxybenzaldehyde in Step 1.
Step 2: Preparation of methyl 3-[2-[2-bromo-4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy late
Figure imgf000091_0001
44b
To a solution of 2-(3-bromo-4-hydroxy-phenyl)-8-chloro-chromen-4-one (250 mg, 711 mmol), methyl 3-(2-(4-(tosyloxy)phenoxy)ethoxy)cyclobutanecarboxylate (299 mg, 711 m mol) in DMF (5 mL) was added K2CO3 (98.3 mg, 711 mihoΐ). The mixture was then stirred at 50°C overnight. After the reaction was completed, to the reaction was added water (30 mL) and the resulting suspension was filtered. The solid was collected and dried in vacuo to give methyl 3- [2- [2-bromo-4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylate (200 mg, 55.4%) as a yellow solid. MS obsd. (ESI+) [(M+H)+]: 507.1.
Step 3: Preparation of ,v-3-[2-[2-bronio-4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000091_0002
44
To a solution of methyl 3-[2-[2-bromo-4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (200 mg, 394 m mol) in the mixed solvent of MeOH (5 mL) and water (1 mL) was added LiOH (40.9 mg, 1.71 mmol). The mixture was then stirred at room temperature for 4 hours. After the reaction was completed, the mixture was concentrated in vacuo and the residue was purified by prepare HPLC to give cis configuration of 3- [2- [2- bromo-4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid ( 90 mg, 32%) as a white foam, the trans isomer was not collected in the purification. 1 H NMR (DMSO- d6, 400MHz): d ppm 12.0-12.4 (m, 1H), 8.1-8.2 (m, 1H), 8.05-8.12 (m, 1H), 7.98-8.04 (m, 2H),
7.46-7.56 (m, 1H), 7.35-7.41 (m, 1H), 7.15-7.22 (m, 1H), 4.28-4.33 (m, 2H), 3.91-4.08 (m, 1H), 3.66-3.78 (m, 2H), 2.54-2.65 (m, 1H), 2.38-2.46 (m, 2H), 1.91-2.05 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 493.1.
Example 45
Ci.s-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyclopropyl- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000092_0001
45
Step 1: Preparation of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyclopropyl- phenoxy ] ethoxy] cyclobutanecarboxy late
Figure imgf000092_0002
45a
A mixture methyl 3-[2-[2-bromo-4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (200 mg, 394 m mol), cyclopropylboronic acid (67.7 mg, 788 m mol), K3PO4 (251 mg, 1.18 mmol) and Bis(triphenylphosphine)palladium(II) chloride (55.3 mg, 78.8 mihoΐ) in the mixed solvent of dioxane (5 mL) and water (2 mL) was heated at 100 °C under microwave radiation for 2 hours. After the reaction was completed, the mixture was diluted with water (15 mL) and the resulting mixture was extracted with EtOAc (20 mL) three times. The combined organic layer was washed with brine, dried over Na2S04 and concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with PE:EA 100:1 ~2:l) to give methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyclopropyl- phenoxy] ethoxy] cyclobutanecarboxylate (150 mg, 81.2%) as a yellow solid. MS obsd. (ESI+) [(M+H)+]: 469.1.
Step 2: Preparation of ,s-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyclopropyl- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000093_0001
To a mixture of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyclopropyl- phenoxy] ethoxy] cyclobutanecarboxylate (150 mg, 320 m mol) in the mixed solvent of MeOH (5 mL) and water (1 mL) was added LiOH (24 mg, 1.0 mmol). The mixture was then stirred at room temperature for 4 hours. After the reaction was completed, the mixture was concentrated in vacuo and the residue was purified by prepare HPLC to give cA-3-[2-[4-(8-chloro-4-oxo- chromen-2-yl)-2-cyclopropyl-phenoxy]ethoxy]cyclobutanecarboxylic acid (11 mg, 11.6%) as a white foam, the trans- isomer was not collected in the purification. 1 H NMR (DMSO-rfe,
400MHz): d ppm 12.05-12.18 (m, 1H), 7.87-8.05 (m, 3H), 7.41-7.59 (m, 2H), 7.10-7.23 (m, 2H), 4.18-4.28 (m, 2H), 3.94-4.03 (m, 1H), 3.67-3.75 (m, 2H), 2.56-2.65 (m, 2H), 2.38-2.45 (m, 2H), 2.14-2.24 (m, 1H), 1.94-2.08 (m, 1H), 0.96-1.03 (m, 2H), 0.79-0.87 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 455.1.
Example 46
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyano-phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000094_0001
46
Step 1: Preparation of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyano- phenoxy ] ethoxy] cyclobutanecarboxy late
Figure imgf000094_0002
46a
A mixture of methyl 3-[2-[2-bromo-4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (150 mg, 295 m mol) and copper (I) cyanide (106 mg, 1.18 mmol) in NMP (4 mL) was heated at 160 °C under microwave radiation for 1.5 hours. After the reaction was completed, the mixture was diluted with EtOAc (50 mL). The resulting suspension was filtred through silca pad and the filtrate was concentrate in vacuo to give the crude methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyano- phenoxy] ethoxy] cyclobutanecarboxylate (134 mg, 100%) as a brown oil, which was used in next step directly without further purification. Step 2: Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyano- phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000095_0001
46 To a solution of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyano- phenoxy] ethoxy] cyclobutanecarboxy late (130 mg, 286 m mol) in the mixed solvent of MeOH (10 mL) and water (2 mL) was added LiOH (100 mg, 4.18 mmol). After the reaction was completed, the mixture was quenched with AcOH (0.3g) and the resulting mixture was purified by
preparative HPLC to give 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyano- phenoxy] ethoxy] cyclobutanecarboxy lie acid (22mg, 17.1%) as a white solid. 1 H NMR (DMSO- d6, 400MHz): d ppm 8.47 - 8.38 (m, 1H), 8.35 - 8.27 (m, 1H), 7.99 - 7.90 (m, 2H), 7.50 - 7.38 (m, 2H), 7.19 - 7.12 (m, 1H), 4.36 - 4.24 (m, 2H), 3.98 - 3.86 (m, 1H), 3.68 - 3.61 (m, 2H), 2.48 (m, 2H), 2.16 - 2.03 (m, 1H), 1.98 - 1.87 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 440.1.
Example 47 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-methyl-phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000095_0002
47
Step 1: Preparation of 8-chloro-2-(4-hydroxy-3-methyl-phenyl)chromen-4-one
Figure imgf000096_0001
47a
Compound 47a was prepared in analogy to the procedure described for the preparation of compound 3c by using 4-methoxy-3-methyl-benzaldehyde as the starting material instead of 4- methoxybenzaldehyde in Step 1.
Step 2: Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-methyl- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000096_0002
47 Example 47 was prepared in analogy to the procedure described for the preparation of
Example 15 by using 8-chloro-2-(4-hydroxy-3-methyl-phenyl)chromen-4-one as the starting material instead of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one in Step 5.
Example 47: lH NMR (DMSC )-d6, 400MHz): d ppm 12.27 - 11.83 (m, 1H), 8.03 - 7.91 (m, 4H), 7.49 (t, 7=7.8 Hz, 1H), 7.16 (d, 7=8.6 Hz, 1H), 7.05 (s, 1H), 4.22 - 4.16 (m, 2H), 3.98 (t, 7=7.0 Hz, 1H), 3.72 - 3.66 (m, 2H), 2.58 (d, 7=8.1 Hz, 1H), 2.47 - 2.38 (m, 2H), 2.26 (s, 3H),
2.05 - 1.92 (m, 2H) MS obsd. (ESE) [(M+H)+]:429.2.
Example 48
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-isopropyl-phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000097_0001
48
Step 1: Preparation of 8-chloro-2-(4-hydroxy-3-isopropyl-phenyl)chromen-4-one
Figure imgf000097_0002
Compound 48a was prepared in analogy to the procedure described for the preparation of compound 3c by using 3-isopropyl-4-methoxy-benzaldehyde as the starting material instead of 4-methoxybenzaldehyde in Step 1.
Step 2: Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-isopropyl- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000097_0003
48
Example 48 was prepared in analogy to the procedure described for the preparation of Example 15 by using 8-chloro-2-(4-hydroxy-3-isopropyl-phenyl)chromen-4-one as the starting material instead of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one in Step 5. Example 48:
Figure imgf000098_0001
NMR (DMSO -d6, 400MHz): d ppm 8.04 - 7.91 (m, 4H), 7.53 - 7.43 (m,
1H), 7.22 - 7.14 (m, 1H), 7.13 - 7.05 (m, 1H), 4.25 - 4.16 (m, 2H), 3.98 - 3.89 (m, 1H), 3.71 - 3.65 (m, 2H), 2.90 - 2.80 (m, 1H), 2.61 - 2.57 (m, 0.6 H), 2.44 - 2.33 (m, 2H), 2.16 - 2.07 (m, 0.4 H), 2.03 - 1.92 (m, 2H), 1.34 - 1.17 (m, 6H). MS obsd. (ESI+) [(M+H)+]:457.3.
Example 50
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-methoxy-phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000098_0002
50 Step 1: Preparation of 8-chloro-2-(2-fluoro-4-hydroxy-phenyl)chromen-4-one
Figure imgf000098_0003
50a
Compound 49a was prepared in analogy to the procedure described for the preparation of compound 3c by using 2-fluoro-4-methoxy-benzaldehydeas the starting material instead of 4- methoxybenzaldehyde in Step 1.
Step 2: Preparation of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-fluoro- phenoxy ] ethoxy] cyclobutanecarboxy late
Figure imgf000099_0001
50b
Compound 49b was prepared in analogy to the procedure described for the preparation of compound 15f by using 8-chloro-2-(2-fluoro-4-hydroxy-phenyl)chromen-4-one as the starting material instead of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one in Step 5.
Step 2: Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-methoxy- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000099_0002
50
To a mixture of methyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-fluoro- phenoxy] ethoxy] cyclobutanecarboxy late (120 mg, crude, 60% purity) in DMF (3 mL) was added LiOH (48 mg, 2 mmol), MeOH (10 mL) and water (1 mL) and the mixture was then stirred at room temperature for 72 hours. The mixture was then filtered and the filtrate was concentrated in vacuo, the residue was purified by preparative HPLC to give 3-[2-[4-(8-chloro-4-oxo-chromen- 2-yl)-3-methoxy-phenoxy]ethoxy]cyclobutanecarboxylic acid (20 mg, 10.3%) as a white solid.
Example 50:
Figure imgf000099_0003
NMR (DMSC )-d6, 400MHz): d ppm 12.19 (s, 1H), 7.98 (d, 7=7.5 Hz, 3H),
7.47 (s, 1H), 7.05 (s, 1H), 6.81 (s, 2H), 4.26 - 4.19 (m, 2H), 3.97 (s, 3H), 3.95 - 3.90 (m, 1H), 3.70 - 3.64 (m, 2H), 2.63 - 2.55 (m, 0.7 H), 2.47 - 2.40 (m, 2H), 2.22 - 2.13 (m, 0.3 H), 2.04 - 1.96 (m, 2H). MS obsd. (ESL) [(M+H)+]:445.2.
Example 51 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-ethoxy-phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000100_0001
51 Step 1: Preparation of (/i)- l-(3-chloro-2-hydroxy-phenyl)-3-(2-ethoxy-4-niethoxy- phenyl)prop-2-en-l-one
Figure imgf000100_0002
51a
To a mixture of 2-fluoro-4-methoxybenzaldehyde (600 mg, 3.89 mmol) and l-(3-chloro-2- hydroxyphenyl)ethanone (664 mg, 3.89 mmol) in EtOH (25 mL) was added KOH (436 mg, 7.79 mmol) and the mixture was then stirred at 60 °C overnight. The reaction was then adjusted to pH- 4.0 by additon of 2N HC1 and the resulting suspension was filtred. The solid was collected and dried in vacuo to give the crude (£’)-l-(3-chloro-2-hydroxy-phenyl)-3-(2-ethoxy-4-methoxy- phenyl)prop-2-en-l-one (51a) (600mg, 50% purity) as a orange solid, which was used in next step directly without further purification. MS obsd. (ESI+) [(M+H)+]:333.2.
Step 2: Preparation of 8-chloro-2-(2-ethoxy-4-hydroxy-phenyl)chromen-4-one
Figure imgf000100_0003
51b
Compound 51b was prepared in analogy to the procedure described for the preparation of compound 3c by using (£’)-l-(3-chloro-2-hydroxy-phenyl)-3-(2-ethoxy-4-methoxy-phenyl)prop- 2-en-l-one as the starting material instead of (£’)-l-(3-chloro-2-hydroxy-phenyl)-3-(4- methoxyphenyl)prop-2-en-l-one in Step 2.
Step 3: Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-ethoxy- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000101_0001
51 Example 51 was prepared in analogy to the procedure described for the preparation of
Example 15 by using 8-chloro-2-(2-ethoxy-4-hydroxy-phenyl)chromen-4-one as the starting material instead of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one in Step 5.
Example 51: lH NMR (DMSC )-d6, 400MHz): d ppm 7.98 (d, 7=8.1 Hz, 3H), 7.51 - 7.43 (m, 1H), 7.10 (s, 1H), 6.84 - 6.77 (m, 2H), 4.28 - 4.15 (m, 4H), 3.99 - 3.90 (m, 1H), 3.70 - 3.63 (m, 2H), 2.63 - 2.53 (m, 2H), 2.45 - 2.40 (m, 0.5H), 2.22 - 2.11 (m, 0.5H), 2.05 - 1.94 (m, 2H),
1.43 (t, 7=6.9 Hz, 3H). MS obsd. (ESE) [(M+H)+]:459.2.
Example 53
3-[2-[4-(3,8-dichloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000101_0002
53
Step 1: Preparation of ethyl 3-[2-[4-(3,8-dichloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxylate
Figure imgf000102_0001
53a
To a solution of ethyl 3-[2-[4-(8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (120 mg, 271 m mol) and 2,6-dimethylpyridine (29 mg, 0.5 mL, 271 m mol) in dichloromethane (12 mL) was added l-chloropyrrolidine-2,5-dione (109 mg, 813 mihoΐ) and the mixture was then stirred at 50 °C for 48 hours. After the reaction was completed, the mixture was concentrated in vacuo and the residue was purified by column chromatography on silica gel to give ethyl 3-[2-[4-(3,8-dichloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (120 mg, 92.8%) as a yellow solid. MS obsd. (ESI+) [(M+H)+]:477.2.
Step 2: Preparation of 3-[2-[4-(3,8-dichloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000102_0002
53
To a solution of ethyl 3-[2-[4-(3,8-dichloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (120 mg, 251 m mol) in the mixed solvent of THF (5 mL), MeOH (5 mL) and Water (1 mL) was added LiOH (36.1 mg, 1.51 mmol) and the mixture was then stirred at room temperature for 4 hours. After the reaction was completed, to the mixture was added AcOH (120 mg, 2mmol) and the resulting mixture was concentrated in vacuo. The residue was then purified by preparative HPLC to give 3-[2-[4-(3,8-dichloro-4-oxo- chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid (26 mg, 21.9%) as a white solid. 1 H NMR (DMSO-^6, 400MHZ): d ppm 1 12.17 - 11.92 (br, 1H), 8.06 (dt, 7=1.5, 8.2 Hz, 2H), 8.01 - 7.94 (m, 2H), 7.55 (t, 7=7.9 Hz, 1H), 7.23 - 7.17 (m, 2H), 4.26 - 4.16 (m, 2H), 3.95 (q, 7=7.5 Hz, 1H), 3.72 - 3.65 (m, 2H), 2.61 - 2.56 (m, 1H), 2.46 - 2.40 (m, 2H), 2.04 - 1.94 (m, 2H). MS obsd. (ESI+) [(M+H)+]:449. l.
Example 55
Ci,s-3-[2-[4-(8-chloro-3-hydroxy-4-oxo-chromen-2- yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000103_0001
55 Step 1: Preparation of 3-[2-[4-[(E)-3-(3-chloro-2-hydroxy-phenyl)-3-oxo-prop-l- enyl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000103_0002
55a Compound 55a was prepared in analogy to the procedure described for the preparation of compound 19b by using l-(3-chloro-2-hydroxy-phenyl)ethanone as the starting material instead of l-(3-chloro-2,4-dihydroxy-phenyl)ethanone in Step 5.
Step 2: Preparation of cis-3-[2-[4-(8-chloro-3-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000104_0001
55
To a solution of 3-[2-[4-[(£)-3-(3-chloro-2-hydroxy-phenyl)-3-oxo-prop-l- enyl]phenoxy]ethoxy]cyclobutanecarboxylic acid (50 mg, 120 m mol) in Acetone (4 mL) was added 4N NaOH solution(l.5 mL) and H202(l.5mL, 30%) at room temperature and the reaction was then stirred at room temperature for 2hours. After the reaction was completed, the mixture was adjusted to pH~2 by addition of 4N HC1 solution. The solid was collected and further purified by recrystallization (EtOH, 5 mL) to give cA-3-[2-[4-(8-chloro-3-hydroxy-4-oxo- chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid (25 mg, 47.4%) as a yellow solid, the trans isomer was not collected in the purification. ' H NMR (DMSO-c/,, 400MHz): d ppm 12.09
(br. s„ 1H), 9.76 (s, 1H), 8.24 (d, 7=9.0 Hz, 2H), 8.07 (dd, 7=1.5, 7.9 Hz, 1H), 7.98 (dd, 7=1.5, 7.7 Hz, 1H), 7.46 (t, 7=7.9 Hz, 1H), 7.18 (d, 7=9.2 Hz, 2H), 4.21 - 4.14 (m, 2H), 3.95 (s, 1H), 3.71 - 3.64 (m, 2H), 2.63 - 2.54 (m, 1H), 2.47 - 2.40 (m, 2H), 2.04 - 1.95 (m, 2H). MS obsd. (ESL) [(M+H)+]:43l.5.
Example 56
C¾-3-[2-[4-(8-chloro-3-methoxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000105_0001
56
To a solution of c/5'-3-[2-[4-(8-chloro-3-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (50 mg, 116 m mol) and CS2CO3 (94.5 mg, 290 mmol) in DMF (3 mL) was added Mel (41.2 mg, 18.1 pL, 290 mihoΐ) at room temperature and then the mixture was stirred at 40°C for lhour. Then to the mixture was added LiOH solution(lN in water, 0.5 mL) and the mixture was stirred at room temperature for 30 minutes. After the reaction was completed, the mixture was adjusted to pH~2 by addtion of 2N HC1 and the resulting mixture was concentrated in vacuo. The residue was purifed by preparative HPLC to give cA-3-[2-[4-(8-chloro-3-methoxy-4-oxo-chromen-2- yljphenoxy] ethoxy] cyclobutanecarboxylic acid (40mg, 77.5%) as a yellow solid. 1 H NMR
(DMSO-^6, 400MHZ): d ppm 12.17 (br. s„ 1H), 8.15 - 8.10 (m, 2H), 8.02 (ddd, 7=1.5, 7.8, 17.9 Hz, 2H), 7.49 (t, 7=7.9 Hz, 1H), 7.24 - 7.17 (m, 2H), 4.21 - 4.16 (m, 2H), 4.00 - 3.91 (m, 1H), 3.84 (s, 3H), 3.71 - 3.65 (m, 2H), 2.63 - 2.55 (m, 1H), 2.48 - 2.39 (m, 2H), 2.06 - 1.94 (m, 2H). MS obsd. (ESI+) [(M+H)+]:445.2.
Example 57
3-[2-[4-(8-chloro-4-oxo-thiochromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000105_0002
57 Step 1: Preparation of 8-chloro-2-(4-methoxyphenyl)thiochromen-4-one
Figure imgf000106_0001
57a
A mixture of 2-chlorobenzenethiol (2.56 g, 2 mL, 17.7 mmol) and ethyl 3-(4- methoxyphenyl)-3-oxopropanoate (3.93 g, 3.39 mL, 17.7 mmol) in PPA(20mL) was stirred at l20°C overnight. The reaction was quenched by pouring into water (80 mL) and the resulting mixture was extracted with EtOAc (60 mL) three times. The combined organic layers were washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo. The residue was then purified by column chromatography on silica gel (elution with PE:EtOAc= 5: 1) to give 8- chloro-2-(4-methoxyphenyl)thiochromen-4-one (500 mg, 9.1%) as a grey solid. MS obsd. (ESL) [(M+H)+]:303.l.
Step 2: Preparation of 8-chloro-2-(4-hydroxyphenyl)thiochromen-4-one
Figure imgf000106_0002
57b
Compound 57b was prepared in analogy to the procedure described for the preparation of compound 3c by using 8-chloro-2-(4-methoxyphenyl)thiochromen-4-one as the starting material instead of 8-chloro-2-(4-methoxyphenyl)chromen-4-one in Step 3.
Step 3: Preparation of 3-[2-[4-(8-chloro-4-oxo-thiochromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000107_0001
57
Example 57 was prepared in analogy to the procedure described for the preparation of Example 15 by using 8-chloro-2-(4-hydroxyphenyl)thiochromen-4-one as the starting material instead of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one in Step 5.
Example 57: lH NMR (DMSC )-d6, 400MHz): d ppm 12.19 - 12.07 (m, 1H), 8.35 (dd, 7=1.3, 7.9 Hz, 1H), 8.00 (dd, 7=1.2, 7.8 Hz, 1H), 7.87 - 7.79 (m, 2H), 7.68 (t, 7=7.9 Hz, 1H), 7.28 (s, 1H), 7.17 - 7.11 (m, 2H), 4.21 - 4.15 (m, 2H), 4.07 - 3.90 (m, 1H), 3.67 (dd, 7=3.9, 5.4 Hz, 2H), 2.64 - 2.53 (m, 1H), 2.48 - 2.36 (m, 2H), 2.22 - 1.94 (m, 2H). MS obsd. (ESI+)
[(M+H)+]:43l.2.
Example 58
2-[2-[4-(8-chloro-4-oxo-thiochromen-2-yl)phenoxy]ethoxy]acetic acid
Figure imgf000107_0002
Step 1: Preparation of 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]thiochromen-4-one
Figure imgf000107_0003
58a To a solution of 8-chloro-2-(4-hydroxyphenyl)thiochromen-4-one (40 mg, 139 m mol,) and K2CO3 (38.3 mg, 277 pmol) in DMF (3 mL) was added 2-bromoethanol (26 mg, 14.8 pL, 208 m mol) at room temperature and the mixture was then stirred at 80°C overnight. The mixture was then quenched with water (10 mL) and the resulting suspension was filtered. The solid was collected and dried in vacuo to give the crude 8-chloro-2-[4-(2- hydroxyethoxy)phenyl]thiochromen-4-one (40 mg, 85%) as a white solid, which was used in the next step without further purification. MS obsd. (ESI+) [(M+H)+]:333.2.
Step 2: Preparation of 2-[2-[4-(8-chloro-4-oxo-thiochromen-2-yl)phenoxy]ethoxy]acetic acid
Figure imgf000108_0001
Example 58 was prepared in analogy to the procedure described for the preparation of Example 9 by using 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]thiochromen-4-one as the starting material instead of 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one in Step 1.
Example 58: lH NMR (DMSC )-d6, 400MHz): d ppm 12.03 - 11.97 (m, 1H), 8.36 (d, 7=7.8 Hz, 1H), 8.00 (d, 7=7.8 Hz, 1H), 7.84 (d, 7=8.8 Hz, 2H), 7.68 (t, 7=7.9 Hz, 1H), 7.28 (s, 1H), 7.16 (d, 7=8.8 Hz, 2H), 4.25 - 4.20 (m, 2H), 4.08 (s, 2H), 3.88 - 3.84 (m, 2H). MS obsd. (ESI+)
[(M+H)+]:39l. l.
Example 59 2-[2-[4-(8-chloro-3-hydroxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetic acid
Figure imgf000108_0002
59 Step 1: Preparation of 8-chloro-3-hydroxy-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one
Figure imgf000109_0001
To a solution of (£’)-l-(3-chloro-2-hydroxy-phenyl)-3-[4-(2-hydroxyethoxy)phenyl]prop-2- en-l-one (500 mg, 1.57 mmol) and NaOH (3.92 ml, 15.7 mmol) in EtOH (5 ml) was added H2O2 (1.78 g, 1.6 mL, 15.7 mmol) dropwise at room temperature. After the addition, the mixture was stirred at room temperature for additional 2 hours. The mixture was then adjusted to pH ~2 by addition of cone. HC1 and the resulting suspension was then filtered. The solid was collected and dried in vacuo to give 8-chloro-3-hydroxy-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one (340 mg, 61.9%) as a yellow solid. MS obsd. (ESI+) [(M+H)+]:333.l.
Step 2: Preparation of 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]-3- (methoxymethoxy)chromen-4-one
Figure imgf000109_0002
To a solution of 8-chloro-3-hydroxy-2-[4-(2-hydroxyethoxy)phenyl]chromen-4-one (340 mg, 1.02 mmol) and K2CO3 (212 mg, 1.53 mmol) in DMF (8 mL) was added
bromo(methoxy)methane (153 mg, 100 LI L, 1.23 mmol) at room temperature and the mixture was then stirred at room temperature overnight. The reaction was then diluted with water (30 mL) and extracted with EtOAc (30 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo. The residue was then purified by column chromatography on silica gel to give 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]-3- (methoxymethoxy)chromen-4-one (200 mg, 41.6%) as a yellow solid. MS obsd. (ESI+)
[(M+H)+]:377.l. Step 3: Preparation of tert-butyl 2-[2-[4-[8-chloro-3-(methoxymethoxy)-4-oxo-chromen-2- yl] phenoxy ] ethoxy] acetate
Figure imgf000110_0001
59c To a solution of 8-chloro-2-[4-(2-hydroxyethoxy)phenyl]-3-(methoxymethoxy)chromen-4- one (100 mg, 265 pmol) in DMSO (5 mL) was added NaH (31.8 mg, 796 pmol) at room temperature and the mixture was stirred at room temperature for 15 minutes. Then to the resulting mixture was added tert-butyl 2-bromoacetate (155 mg, 796 m mol) and the mixture was stirred at room temperature for 8 hours. After the reaction was completed, the mixture was diluted with water (30 mL) and the mixture was extracted with EtOAc (30 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo to give the crude tert-butyl 2-[2-[4-[8-chloro-3-(methoxymethoxy)-4-oxo-chromen-2- yl]phenoxy] ethoxy] acetate (130 mg, 100%) as a solid, which was used in the next step directly without further purification. MS obsd. (ESL) [(M+H)+]:49l.2.
Step 4: Preparation of 2-[2-[4-(8-chloro-3-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] acetic acid
Figure imgf000110_0002
A mixture of tert-butyl 2-[2-[4-[8-chloro-3-(methoxymethoxy)-4-oxo-chromen-2- yl]phenoxy] ethoxy] acetate (130 mg, 265 m mol) in the mixed solvent of THF(3mL) and cone.
HC1 (3mL) was stirred at room temperature overnight. After the reaction was completed, the reaction mixture was concentrated in vacuo and the residue was purified by preparative HPLC to give 2-[2-[4-(8-chloro-3-hydroxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetic acid (40 mg, -HO-
36.6%) as a yellow solid. XH NMR (DMSO-J^, 400MHz): d ppm 12.55 (br. s., 1H), 9.76 (br. s., 1H), 8.24 (d, J=9.0 Hz, 2H), 8.07 (dd, J=l.5, 8.1 Hz, 1H), 7.98 (dd, J=l.5, 7.6 Hz, 1H), 7.46 (t, J=7.8 Hz, 1H), 7.19 (d, J=9.3 Hz, 2H), 4.27 - 4.18 (m, 2H), 4.11 (s, 2H), 3.92 - 3.81 (m, 2H). MS obsd. (ESI+) [(M+H)+] : 391.1. Example 60
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-morpholino- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000111_0001
60 Step 1: Preparation of 2-(2-bromo-4-hydroxy-phenyl)-8-chloro-chromen-4-one
Figure imgf000111_0002
60a
Compound 60a was prepared in analogy to the procedure described for the preparation of intermediate 3c by using 2-bromo-4-methoxy-benzaldehyde as the starting material instead of 4- methoxybenzaldehyde in Step 1.
Step 2: Preparation of methyl 3-[2-[3-bromo-4-(8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy late
Figure imgf000112_0001
60b
Compound 60 b was prepared in analogy to the procedure described for the preparation of compound 15f by using 2-(2-bromo-4-hydroxy-phenyl)-8-chloro-chromen-4-one as the starting material instead of 8-chloro-2-(4-hydroxyphenyl)chromen-4-one in Step 5.
Step 3: Preparation of 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-morpholino- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000112_0002
60 To a mixture of methyl 3-(2-(3-bromo-4-(8-chloro-4-oxo-4H-chromen-2- yl)phenoxy)ethoxy)cyclobutanecarboxylate (100 mg, 197 m mol), morpholine (51.5 mg, 591 pmol), TEA (159 mg, 220 mΐ, 1.58 mmol) , sodium tert-butoxide (75.7 mg, 788 mihoΐ), 2'- bis(diphenylphosphino)-l,r-binaphthalene (83 mg, 133 mihoΐ) in dioxane (4 mL) was added Pd2(dba)3 (80 mg, 87.4 mmol) and the mixture was then stirred at 110 °C for 2 hours. After the reaction was completed, the mixture was partitioned between EtOAc (15 mL) and water (15 mL), the organic layer was separated out. The aquatic phase was extracted with EtOAc (15 mL) twice. The organic layer was combined, washed with brine, dried over Na2S04 and concentrated in vacuo. The residue was purified by preparative HPLC to give 3-(2-(4-(8-chloro-4-oxo-4H- chromen-2-yl)-3-morpholinophenoxy)ethoxy)cyclobutanecarboxylic acid (9 mg, 9.14 % yield) as a yellow foam. Example 60: NMR (DMSO -d6, 400MHz): d ppm 7.96-8.03 (m, 2H), 7.71-7.75 (m, 1H), 7.45-7.52 (m, 1H), 7.15 (s, 1H), 6.78-6.85 (m, 1H), 6.68-6.74 (m, 1H), 4.12-4.22 (m, 2H), 3.73- 3.95 (m, 1H), 3.61-3.72 (m, 6H), 2.93-3.03 (m, 4H), 2.56-2.63 (m, 1H), 2.36-2.47 (m, 2H), 2.16 (m, 1H), 1.94-2.04 (m, 1H). MS obsd. (ESI+) [(M+H)+]:500. l. Example 61
2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetamide
Figure imgf000113_0001
To a solution of 2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetic acid (example 9, 400.0 mg, 1.07 mmol), triethylamine (0.6 mL, 4.27 mmol) in THF (20 mL) was added HATU (486.99 mg, 1.28 mmol), ammonium chloride (114.2 mg, 2.13 mmol). The mixture was stirred at room temperature for 12 hours. The mixture was then washed by water (15 mL) and concentrated in vacuo. The residue was then triturated in DMF (l5mL) and the suspension was then filtered to give 2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetamide (75 mg, 16.8%) as a yellow solid. XH NMR (DMSO-J^, 400MHz): d ppm 8.10 (d, 7=8.9 Hz, 2H), 8.00 (d, 7=7.9 Hz, 2H), 7.49 (t, 7=7.8 Hz, 1H), 7.27 (d, 7=7.7 Hz, 1H), 7.19 (d, 7=9.0 Hz, 2H), 7.07 (s, 1H), 4.21-4.32 (m, 2H), 3.90 (s, 2H), 3.81-3.86 (m, 2H). MS obsd. (ESI+)
[(M+H)+]:374. l.
Example 62 Methyl 3-[2-[4-[8-chloro-6-fluoro-4-oxo-3-(trifluoromethoxy)chromen-2- yl] phenoxy ] ethoxy] cyclobutanecarboxy late
Figure imgf000114_0001
62
Step 1: Preparation of methyl 3-[2-[4-(8-chloro-6-fluoro-4-oxo-chromen-2- yl)phenoxy]ethoxy]cyclobutanecarboxylate
Figure imgf000114_0002
62a
Compound 62a was prepared in analogy to the procedure described for the preparation of compound 15f by using l-(3-chloro-5-fluoro-2-hydroxy-phenyl)ethanone as the starting materials instead of l-(3-chloro-4-fluoro-2-hydroxy-phenyl)ethanone in Step 3.
Step 2: Preparation of methyl 3-[2-[4-(3-bromo-8-chloro-6-fluoro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy late
Figure imgf000114_0003
62b
To a mixture of methyl 3-[2-[4-(8-chloro-6-fluoro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (0.6 g, 1.34 mmol) in DCM (10 mL) was added Py (1 mL) and PyHBn (2.15 g, 6.71 mmol) and the mixture was stirred at room temperature for 16 hours. The reaction was quenched with water (30 mL) and the resulting mixture was extracted with DCM (50mL) twice. The combined organic phase was dried over MgS04 and concentrated in vacuo to give the crude of methyl 3-[2-[4-(3-bromo-8-chloro-6-fluoro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (0.5 g, 70.8% ) as a yellow solid, which was used in the next step directly. MS obsd. (ESI+) [(M+H)+]: 525.8.
Step 3: Preparation of 3-[2-[4-(8-chloro-6-fluoro-3-hydroxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000115_0001
To a solution of methyl 3-[2-[4-(3-bromo-8-chloro-6-fluoro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (500 mg, 0.95 mmol) in DMF (5 mL) was added NaOH solution (4N, 5 mL) and the mixture was then stirred at 80 °C for 2 hours. The mixture was poured into water (30 mL) and adjusted to pH=5~6 by addition of cone. HC1. The resulting mixture was extracted with EtOAc (50 mL) twice and the combined organic phase was dried over Na2S04, concentrated in vacuo to give the crude of 3-[2-[4-(8-chloro-6-fluoro-3-hydroxy-4- oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid (0.3 g , 68.5% yield) as a yellow solid, which was used in the next step directly. MS obsd. (ESI+) [(M+H)+]: 449.1.
Step 4: Preparation of methyl 3-[2-[4-(8-chloro-6-fluoro-3-hydroxy-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxylate
Figure imgf000115_0002
62d A solution of 3-[2-[4-(8-chloro-6-fluoro-3-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (0.3 g, 0.65 mmol ) dissolved in hydrogen chloride methanol solution (4 mol/L, 7.5 ml, 30 mmol) was stirred at room temperature for 12 hours. After the reaction was completed, the mixture was concentrated in vacuo to give methyl 3-[2-[4-(8-chloro-6-fluoro-3-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate ( 0.3g, 100% yield) as a white solid, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 463.2.
Step 5: Preparation of methyl 3-[2-[4-[3-[bromo(difluoro)methoxy]-8-chloro-6-fluoro-4- oxo-chromen-2-yl]phenoxy]ethoxy]cyclobutanecarboxylate
Figure imgf000116_0001
62e
To a solution of methyl 3-[2-[4-(8-chloro-6-fluoro-3-hydroxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (200 mg, 0.43 mmol) in NMP (5 mL) was added sodium hydride (34.57 mg, 0.860 mmol) at 0°C, the mixture was then stirred at 0°C for 30 minutes. Then to the resulting mixture was added dibromo(difluoro)methane (0.075 mL,0.86 mmol, 2 eq) and the mixture was stirred at room temperature for 36 hours. The mixture was quenched with saturated NH4Cl solution (5 mL) and extracted with EtOAc (10 mL) three times. The organic phase was combined, washed with brine, dried over Na2S04 and concentrated in vacuo. The residue was re-dissolved in MeOH (2 mL) and the mixture was then filtered. The filtrate was concentrated in vacuo to give the crude of methyl 3-[2-[4-[3- [bromo(difluoro)methoxy]-8-chloro-6-fluoro-4-oxo-chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylate (20 mg, 8.01%) as yellow solid. MS obsd. (ESI+) [(M+H)+]: 592.1.
Step 6: Preparation of methyl 3-[2-[4-[8-chloro-6-fluoro-4-oxo-3- (trifluoromethoxy)chromen-2-yl]phenoxy]ethoxy]cyclobutanecarboxylate
Figure imgf000117_0001
62
To a solution of methyl 3-[2-[4-[3-[bromo(difluoro)methoxy]-8-chloro-6-fluoro-4-oxo- chromen-2-yl]phenoxy]ethoxy]cyclobutanecarboxylate (40.0 mg, 0.070 mmol, 1 eq) in DCM (2 mL) cooled at -70°C was added argentio(tetrafluoro)boron (32.9 mg, 0.170 mmol, 2.5 eq) and the mixture was stirred at 15 °C for l2h ours. The mixture was filtered and the filtrate was concentrated in vacuo to give the crude of methyl 3-[2-[4-[8-chloro-6-fluoro-4-oxo-3- (trifluoromethoxy)chromen-2-yl]phenoxy] ethoxy] cyclobutanecarboxylate, which was further purified by Prep-HPLC to give two diastereomers with cis- and trans- configuration, one of which is characterized as Example 62-A (4.8 mg, 12.8%) and the other is Example 62-B (5.2 mg, 14.3%). as white solid.
Example 62-A: lH NMR (CDCb, 400MHz): d ppm 8.15 (d, 7=9.03 Hz, 2 H), 7.31 - 7.40 (m, 2 H), 7.06 (d, 7=8.91 Hz, 2 H), 4.32 (t, 7=6.78 Hz, 1 H), 4.19 - 4.26 (m, 2 H), 3.74 - 3.83 (m,
2 H), 3.71 (s, 3 H), 3.01 - 3.13 (m, 1 H), 2.54-2.57 (m, 2 H), 2.30 - 2.34 (m, 2 H). MS obsd.
(ESr) [(M+H)+]: 531.1.
Example 62-B: lH NMR (CDCb, 400MHz): d ppm 8.15 (d, 7=9.03 Hz, 2 H), 7.30-7.38 (m, 2 H), 7.06 (d, 7=8.93 Hz, 2 H), 4.22 (t, 7=4.65Hz, 1 H), 4.02(m, 2H), 3.73-3.83 (m, 2 H), 3.70 (s,3 H), 2.60-2.75 (m, 1H), 2.55-2.65 (m, 2H), 2.24-2.36 (m, 2 H) . MS obsd. (ESI+) [(M+H)+]: 531.1.
Example 63
3-[2-[4-[8-chloro-6-fluoro-4-oxo-3-(trifluoromethoxy)chromen-2- yl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000118_0001
63
To a mixture of methyl 3-[2-[4-[8-chloro-6-fluoro-4-oxo-3-(trifluoromethoxy)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylate (100 mg, 0.19 mmol) in the mixed solvent of THF (5mL) and water (2 mL) was added lithium hydroxide (48 mg, 2 mmol) and the mixture was then stirred at room temperature for 12 hours. The mixture was adjusted to pH~4 by addition of 1M HC1 and extracted with EtOAc (10 mL) three times. The combined organic layer was washed with brine, dried over Na2S04 and concentrated in vacuo. The residue was purified by preparative HPLC to give 3-[2-[4-[8-chloro-6-fluoro-4-oxo-3-(trifluoromethoxy)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylic acid (17.3 mg, 17.7% yield) as a white solid. 1 H NMR (DMSO -d6, 400MHz): d ppm 8.07(d, 7=9.2 Hz, 2 H), 7.90-7.93 (m, 1 H), 7.77-7.79 (m, 1H), 7. l9(d, 7=8.8 Hz, 2 H), 4.20-4.32 (m, 2H), 3.90-4.15 (m, 1 H), 3.67-3.69 (m, 2 H), 2.80-2.99 (m, 1H), 2.40-2.50 (m, 2H), 2.11-2.23 (m, 1 H), 1.95-2.04 (m, 1H) . MS obsd. (ESI+) [(M+H)+]: 517.1. Example 64
3-[2-[4-(3-bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000118_0002
64 Step 1: Preparation of ethyl 3-[2-[4-(3-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy]ethoxy]cyclobutanecarboxylate
Figure imgf000119_0001
64a To a solution of ethyl 3-(2-(4-(8-chloro-4-oxo-4H-chromen-2- yl)phenoxy)ethoxy)cyclobutanecarboxylate (200 mg, 452 m mol) in DCM (5 mL) was added 1- bromopyrrolidine-2,5-dione (88.4 mg, 497 m mol), the mixture was then stirred at room temperature for 5 hours. The mixture was then concentrated in vacuo and purified by column chromatography on silica gel (elution with DCM:MeOH 100:1-10:1) to give ethyl 3-[2-[4-(3- bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylate (30 mg, 12.7% yield) as a yellow solid. MS obsd. (ESI+) [(M+H)+]: 520.9.
Step 2: Preparation of 3-[2-[4-(3-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000119_0002
64
To a solution of ethyl 3-(2-(4-(3-bromo-8-chloro-4-oxo-4H-chromen-2- yl)phenoxy)ethoxy)cyclobutanecarboxylate (30 mg, 57.5 m mol) in the mixed solvent of THF (1 mL) and H20 (0.5 mL) was added LiOH (6.88 mg, 287 m mol) and the mixture was then stirred at room temperature for 1 hour. After the reaction was completed, the mixture was adjusted to pH~6 by addition of HC1 (1 N) and concentrated in vacuo. The residue was purified by preparative HPLC to give 3-[2-[4-(3-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (3.6 mg, 12.6%) as a light yellow solid. 1 H NMR (DMSO-rfc, 400MHz): d ppm 8.06 (m, 2H), 7.93 (d, 7=9.05 Hz, 2H), 7.55 (s, 1H), 7.19 (d, 7=9.05 Hz, 2H), 4.15-4.24 (m, 2H), 3.91-4.02 (m, 1H), 3.60-3.72 (m, 2H), 2.56-2.63 (m, 1H),
2.40-2.51 (m, 2H), 1.90-2.08 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 492.9.
Example 65
3-[2-[4-(3-benzyloxy-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000120_0001
65
To a solution of 3-(2-(4-(8-chloro-3-hydroxy-4-oxo-4H-chromen-2- yl)phenoxy)ethoxy)cyclobutanecarboxylic acid (30 mg, 69.6 m mol) and Cs2C03 (56.7 mg, 174 mmol) in DMF (3 mL) was added (bromomethyl)benzene (26.2 mg, 153 mihoΐ) and the mixture was then stirred at room temperature overnight. Then to the reaction was added LiOH(24 mg, lmmol) and the mixture was stirred at 50 °C for 1 hour. The mixture was adjusted to pH~6 by addition of HC1 (1 N) and concentrated in vacuo. The residue was purified by preparative HPLC to give 3-[2-[4-(3-benzyloxy-8-chloro-4-oxo-chromen-2- yljphenoxy] ethoxy] cyclobutanecarboxylic acid (18 mg, 45.7%) as a light yellow solid. 1 H NMR(DMSO- d6, 400MHz): d ppm 7.93-8.14 (m, 4H), 7.50 (s, 1H), 7.28-7.44 (m, 5H), 7.14 (d, 7=9.05 Hz, 2H), 5.11 (s, 2H), 4.15-4.24 (m, 2H), 3.90-3.99 (m, 1H), 3.57-3.73 (m, 2H), 2.55- 2.63 (m, 1H), 2.40-2.51 (m, 2H), 1.88-2.09 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 521.1.
Example 66 3-[2-[4-[8-chloro-4-oxo-6-(trifluoromethyl)chromen-2- yl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000121_0001
66 Step 1: Preparation of 6-bromo-8-chloro-2-(4-hydroxyphenyl)chromen-4-one
Figure imgf000121_0002
To a solution of 6-bromo-8-chloro-2-(4-methoxyphenyl)chromen-4-one (intermediate 22b, 5.0 g, 13.68 mmol) in chloroform (20 mL) cooled at 0 °C was added BBr3 (17.13 g, 68.3 mmol). The mixture was then stirred at room temperature for 16 hours. After the reaction was completed, the reaction mixture was quenched with water (50 mL) and the resulting suspension was filtered and the solid was washed by water (200mL) and EtOH (100 mL) in sequence. The solid was concentrated in vacuo to give the crude product of 6-bromo-8-chloro-2-(4- hydroxyphenyl)chromen-4-one (4.3 g, 71.55% yield) as a yellow solid, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 351.6.
Step 2: Preparation of methyl 3-[2-[4-(6-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy late
Figure imgf000122_0001
66b
To a solution of 6-bromo-8-chloro-2-(4-hydroxyphenyl)chromen-4-one (4.3 g, 13.56 mmol) and potassium carbonate (3.75 g, 27.12 mmol) in DMF (30 mL) was added methyl 3-[2-(p- tolylsulfonyloxy)ethoxy]cyclobutanecarboxylate (int-2. 4.45 g, 13.56 mmol) and the mixture was then stirred at 80 °C for 16 hours. The reaction mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by column chromatography on silica gel (eluent DCM: MeOH=30: 1-10:1) to give the methyl 3-[2-[4-(6-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (4.5 g, 63.11% yield) as a yellow solid. MS obsd. (ESI+) [(M+H)+]: 507.8.
Step 3: Preparation of methyl 3-[2-[4-[8-chloro-4-oxo-6-(trifluoromethyl)chromen-2- yl] phenoxy ] ethoxy] cyclobutanecarboxylate
Figure imgf000122_0002
66c To a solution of copper(I) iodide (0.13 mL, 3.94 mmol) and sodium trifluoro acetate (1.07 g,
7.88 mmol) in NMP (10 mL) was added methyl 3-[2-[4-(6-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (400 mg, 0.79 mmol) and the mixture was stirred at 160 °C under nitrogen atmosphere for 4 hours. After the reaction was completed, the mixture was filtered and the filtrate was purified by preparative HPLC to give the crude of methyl 3- [2- [4-[8-chloro-4-oxo-6-(trifluoromethyl)chromen-2-yl]phenoxy]ethoxy]cyclobutanecarboxylate (230 mg, 52.89% yield) as brown oil. MS obsd. (ESI+) [(M+H)+]: 496.8.
Step 4: Preparation of 3-[2-[4-[8-chloro-4-oxo-6-(trifluoromethyl)chromen-2- yl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000123_0001
66
To a solution of methyl 3-[2-[4-[8-chloro-4-oxo-6-(trifluoromethyl)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylate (230 mg, 0.46 mmol, 1 eq) in the mixed solvent of THF (3 mL), methanol (3 mL) and water (3 mL) was added lithium hydroxide (0.03 mL, 3.32 mmol) and the mixture was then stirred at room temperature for 2 hours. After the reaction was completed, the mixture was concentrated in vacuo and the residue was purified by preparative HPLC to give [2-[4-[8-chloro-4-oxo-6-(trifluoromethyl)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylic acid (150 mg, 44.44% yield) as a yellow solid. The solid was further purified by supercritical fluid chromatography (SFC) to give two diastereomers with cis- and trans- configuration, one of which is characterized as Example 66-A (55.3 mg, 36.2%) and the other is Example 66-B (54.6 mg, 34.1%). as white solid.
Example 66-A: lH NMR (DMSO -d6, 400MHz): d ppm 8.41 (m, 1H), 8.15 (m 1H), 8.04 (d, J=8.2 Hz, 2H), 7.18-7.07 (m, 3H), 4.24 - 4.15 (m, 3H), 3.65 (m, 2H), 2.82 (t, 7=10.0 Hz, 1H), 2.43 - 2.31 (m, 2H), 2.14 - 2.05 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 482.9.
Example 66-B: lH NMR (DMSO -d6, 400MHz): d ppm 8.39 (m, 1H), 8.13 (m, 1H), 8.02 (d, 7=7.9 Hz, 2H), 7.17-7.07 (m, 3H), 4.16 (m, 2H), 3.96 - 3.84 (m, 1H), 3.65 (br s, 2H), 2.45 - 2.30 (m, 3H), 2.06 - 1.95 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 482.9.
Example 67
3-[2-[4-(8-chloro-6-cyclopropyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000124_0001
67
Step 1: Preparation of methyl 3-[2-[4-(8-chloro-6-cyclopropyl-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy late
Figure imgf000124_0002
67a
To a solution of methyl 3-[2-[4-(6-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (500 mg, 0.980 mmol), tricyclo hexyl phosphine (27.61 mg, 0.100 mmol), cyclopropylboronic acid (110 mg, 1.28 mmol) and phosphoric acid, potassium salt (0.24 mL, 2.95 mmol) in the mixed solvent of toluene (10 mL) and water (1 mL) was added palladium (II) acetate (11.0 mg, 0.050 mmol). The mixture was then stirred at 100 °C for 12 hours under N2 atmosphere. After the reaction was completed, the mixture was concentrated in vacuo and the residue was purified by column chromatography on silica gel (elution with PE:EtOAc 2:1) to give methyl 3-[2-[4-(8-chloro-6-cyclopropyl-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (300 mg, 58.47% yield) as a yellow oil.
Step 2: Preparation of 3-[2-[4-(8-chloro-6-cyclopropyl-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000125_0001
67
To a solution of methyl 3-[2-[4-(8-chloro-6-cyclopropyl-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (400 mg, 0.85 mmol) in THF (5 mL)/methanol (5 mL)/water (1 mL) was added lithium hydroxide (0.04 mL, 4.27 mmol). The mixture was stirred at room temperature for 2 hours. After the reaction was completed, the mixture was concentrated in vacuo and the residue was purified by preparative HPLC to give 3-[2-[4-(8-chloro-6- cyclopropyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid (119.5 mg, 30.74% yield) as a white solid. XH NMR (DMSO-^fo, 400MHz): d ppm 12.19 (br s, 1H), 8.08 - 8.00 (m, 2H), 7.69 (s, 1H), 7.64 (s, 1H), 7.15 (d, 7=8.5 Hz, 2H), 7.00 (d, 7=1.4 Hz, 1H), 4.22 -
3.91 (m, 3H), 3.72 - 3.62 (m, 2H), 2.97 - 2.56 (m, 1H), 2.47 - 2.32 (m, 2H), 2.22 - 1.94 (m, 3H), 1.08 - 0.99 (m, 2H), 0.84 - 0.75 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 455.0.
Example 68
3-[2-[4-[8-chloro-4-oxo-6-(2-oxopyrrolidin-l-yl)chromen-2- yl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000125_0002
68
Step 1: Preparation of methyl 3-[2-[4-[8-chloro-4-oxo-6-(2-oxopyrrolidin-l-yl)chromen-2- yl] phenoxy] ethoxy] cyclobutanecarboxylate
Figure imgf000126_0001
68a
To a mixture of methyl 3-[2-[4-(6-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate (300 mg, 0.59 mmol), 2-pyrrolidone (0.8 mL, 10.58 mmol), cesium carbonate (192.5 mg, 0.59 mmol) in l,4-dioxane (10 mL) was added tBuXPhos PD G3 (46.9 mg, 0.060 mmol) and the reaction was then stirred at 90 °C for 12 hours under N2 atmosphere. After the reaction was completed, the mixture was filtered and filtrate was concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with PE:EtOAc 10:1-2:1) to give methyl 3-[2-[4-[8-chloro-4-oxo-6-(2-oxopyrrolidin-l- yl)chromen-2-yl]phenoxy]ethoxy]cyclobutanecarboxylate (130 mg, 17.19% yield) as a yellow solid. MS obsd. (ESE) [(M+H)+]: 511.9.
Step 2: Preparation of 3-[2-[4-[8-chloro-4-oxo-6-(2-oxopyrrolidin-l-yl)chromen-2- yl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000126_0002
68
To a solution of methyl 3-[2-[4-[8-chloro-4-oxo-6-(2-oxopyrrolidin-l-yl)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylate (110 mg, 0.21 mmol) in the mixed solvent of THF (6 mL) and water (2 mL) was added lithium hydroxide (25.6 mg, 1.07 mmol) and the mixture was stirred at room temperature for 12 hours. After the reaction was completed, the mixture was adjusted to pH=6 by addition of AcOH and the resulting mixture was concentrated in vacuo, the residue was purified by preparative HPLC to give the 3-[2-[4-[8-chloro-4-oxo-6-(2- oxopyrrolidin-l-yl)chromen-2-yl]phenoxy]ethoxy]cyclobutanecarboxylic acid (27 mg, 0.050 mmol, 25.24% yield) as a yellow solid. CH NMR (DMSO-J^, 400MHz): d ppm 12.2 (br s, 1H), 8.44 (s, 1H), 8.06 - 8.09 (m, 3H), 7.16 - 7.18 (m, 2H), 7.04 (s, 1H), 4.12-4.20 (m, 2H), 3.90-3.96
(m, 2H), 3.66-3.69 (m, 2H), 2.65-2.86 (m, 1H), 2.51-2.58 (m, 3H), 2.28-2.41 (m, 2H), 1.90-2.15 (m, 4H). MS obsd. (ESI+) [(M+H)+]: 498.1.
Example 69
3-[2-[4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2- yl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000127_0001
69
Step 1: Preparation of 7-bromo-8-chloro-2-(4-hydroxyphenyl)chromen-4-one
Figure imgf000127_0002
Compound 69a was prepared in analogy to the procedure described for the preparation of compound 66a by using 7-bromo-8-chloro-2-(4-methoxyphenyl)chromen-4-one as the starting materials instead of 6-bromo-8-chloro-2-(4-methoxyphenyl)chromen-4-one in Step 1.
Step 2: Preparation of methyl 3-[2-[4-(7-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] cyclobutanecarboxy late
Figure imgf000128_0001
69b
Compound 69b was prepared in analogy to the procedure described for the preparation of compound 66b by using 7-bromo-8-chloro-2-(4-hydroxyphenyl)chromen-4-one as the starting materials instead of 6-bromo-8-chloro-2-(4-hydroxyphenyl)chromen-4-one in Step 2.
Step 3: Preparation of 3-[2-[4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2- yl] phenoxy ] ethoxy] cyclobutanecarboxylic acid
Figure imgf000128_0002
69 Example 69 was prepared in analogy to the procedure described for the preparation of example 66 by using methyl 3-[2-[4-(7-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate as the starting materials instead of methyl 3-[2-[4-(6- bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylate in Step 3.
Example 69: lH NMR (DMSC )-d6, 400MHz): d ppm 8.05-8.17 (m, 3H), 7.83-7.95 (m, 1H), 7.14-7.23 (m, 3H), 4.13-4.25 (m, 3H), 3.62-3.71 (m, 2H), 2.87-2.97 (m, 1H), 2.39 (m, 2H),
2.10-2.22 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 482.9.
Example 70
3-[2-[4-(8-chloro-7-cyclopropyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid
Figure imgf000129_0001
70
Example 70 was prepared in analogy to the procedure described for the preparation of example 67 by using methyl 3-[2-[4-(7-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylate as the starting materials instead of methyl 3-[2-[4-(6- bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylate in Step l.The solid of 3-[2-[4-(8-chloro-7-cyclopropyl-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid (l30mg, mmol) was further purified by supercritical fluid chromatography (SFC) to give two diastereomers with cis- and trans- configuration, one of which is characterized as Example 70-A (45 mg, 17.8%) and the other is Example 70-B (30 mg, 27.7%) as white solid.
Example 70: lH NMR (DMSC )-d6, 400MHz): d ppm 12.19 (br s, 1H), 8.09 (d, 7=8.9 Hz, 2H), 7.90 - 7.83 (m, 1H), 7.18 (d, 7=8.9 Hz, 2H), 7.08 (d, 7=8.4 Hz, 1H), 7.02 (s, 1H), 4.14 - 4.22 (m, 2H), 3.87-4.11 (m, 1H), 3.64 - 3.7l(m, 2H), 2.86 - 2.96 (m, 1H), 2.56 - 2.67 (m, 1H), 2.37 - 2.43 (m, 2H), 2.12 - 2.20 (m, 1H), 1.95 - 2.05 (m, 1H), 1.24 - 1.14 (m, 2H), 0.95 - 0.86 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 455.1.
Example 70-A: lH NMR (DMSO-76, 400MHz): d ppm 8.18 - 8.02 (m, 2H), 7.77-7.95(m, 1H), 7.17 (d, 7=8.9 Hz, 2H), 7.08 (d, 7=8.4 Hz, 1H), 7.01 (s, 1H), 4.12-4.24 (m, 3H), 3.63-3.72 (m, 2H), 2.88-2.92 (m, 1H), 2.35-2.43 (m, 3H), 2.11 - 2.21 (m, 2H), 1.15- 1.22 (m, 2H), 0.87 - 0.93 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 455.1.
Example 70-B: lH NMR (DMSO-76, 400MHz): d ppm 8.10 (d, 7=8.9 Hz, 2H), 7.87 (d, 7=8.4 Hz, 1H), 7.18 (d, 7=9.0 Hz, 2H), 7.09 (d, 7=8.4 Hz, 1H), 7.02 (s, 1H), 4.13-4.23 (m, 2H), 3.89-4.03 (m, 1H), 3.63 - 3.73 (m, 2H), 2.55-2.62 (m, 1H), 2.36-2.44 (m, 3H), 1.90-2.05 (m,
2H), 1.13-1.24 (m, 2H), 0.87 - 0.95 (m, 2H). MS obsd. (ESI+) [(M+H)+]: 455.1.
Example 71 ethyl 2-[2-[4-(7-bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetate
Figure imgf000130_0001
71
Step 1: Preparation of ethyl 2-(2-hydroxyethoxy)acetate
Figure imgf000130_0002
71a
To a solution of ethyl 2-(2-benzyloxyethoxy)acetate (2.0 g, 8.39 mmol) in ethanol (20 mL) was added palladium hydroxide (1.18 g, 8.39 mmol) and the mixture was then hydrogenated under H2 atmosphere at room temperature overnight. After the reaction was completed, the reaction was filtered through silica gel pad and the filtrate was concentrated in vacuo to give ethyl 2-(2-hydroxyethoxy)acetate (1.1 g, 7.42 mmol, 44.23% yield) as a colorless oil.
Step 2: Preparation of ethyl 2-[2-(p-tolylsulfonyloxy)ethoxy]acetate
Figure imgf000130_0003
71b
To a solution of ethyl 2-(2-hydroxyethoxy)acetate (1.1 g, 7.42 mmol) in DCM (20 mL) at room temperature was added triethylamine (2.07 mL, 14.85 mmol), m-toluenesulfonyl chloride (1.7 g, 8.91 mmol) and the reaction mixture was stirred at room temperature for 12 hours. After the reaction was completed, the mixture was concentrated in vacuo and the residue was purified by column chromatography on silica gel (elution with PE: EtOAc=50: 1-10:1) to give ethyl 2-[2- (p-tolylsulfonyloxy)ethoxy]acetate (0.400 g, 1.32 mmol) as a colorless oil. MS obsd. (ESI+) [(M+H)+]: 303.1. Step 3: Preparation of ethyl 2-[2-[4-(7-bromo-8-chloro-4-oxo-chromen-2- yl)phenoxy ] ethoxy] acetate
Figure imgf000131_0001
To a mixture of 7-bromo-8-chloro-2-(4-hydroxyphenyl)chromen-4-one (3.0 g, 8.53 mmol) and ethyl 2-[2-(p-tolylsulfonyloxy)ethoxy]acetate (3.1 g, 10.24 mmol) in DMF (30 mL) was added potassium carbonate (1.77 g, 12.8 mmol) and the mixture was stirred at 80 °C for 16 hours. After the reaction was completed, the mixture was poured into water (50mL) and extracted with EtOAc (100 mL) twice. The combined organic phase was washed with brine, dried over Na2S04 and concentrated in vacuo. The residue was then triturated in EtOAc (15 mL) and the mixture was then filtered. The solid was collected and dried in vacuo to give ethyl 2-[2-[4-(7-bromo-8- chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetate (3.5 g, 58.23% yield) as a yellow solid. 1 H NMR (DMSO -d6, 400MHz): d ppm 8.08 (br d, 7=9.05 Hz, 2H), 7.81-7.94 (m, 2H), 7.16 (br d, 7=9.17 Hz, 2H), 7.08 (s, 1H), 4.17-4.29 (m, 4H), 4.06-4.15 (m, 2H), 3.82-3.92 (m, 2H), 1.21 (t, 7=7.15 Hz, 3H). MS obsd. (ESE) [(M+H)+]: 482.1.
Example 72
3-[2-[4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2-yl]-2-methyl- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000131_0002
72 Step 1: Preparation of 4-(methoxymethoxy)-3-methyl-benzaldehyde
Figure imgf000132_0001
72a
To a solution of 4-hydroxy-3-methyl-benzaldehyde (3000.0 mg, 22.03 mmol) and chloro methyl methyl ether (2.01 mL, 26.44 mmol) in THF (40 mL) cooled at 0 °C was added and sodium hydride (634.59 mg, 26.44 mmol), the mixture was stirred at 0 °C for 30 minutes. After the reaction was completed, the mixture was quenched by water (30 mL) and the resulting mixture was extracted with EtOAc (20 mL) twice. The combined organic layer was dried over Na2S04 and concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with PE: EtOAc 10:1- 5:1) to give 4-(methoxymethoxy)-3-methyl- benzaldehyde as a yellow liquid. MS obsd. (ESL) [(M+H)+]: 181.1.
Step 2: Preparation of (/t)- l-(4-bromo-3-chloro-2-hydroxy-phenyl)-3-[4- (methoxymethoxy)-3-methyl-phenyl]prop-2-en-l-one
Figure imgf000132_0002
72b
A mixture of l-(4-bromo-3-chloro-2-hydroxy-phenyl)ethanone (2870.0 mg, 11.5 mmol), potassium hydroxide (3227.3 mg, 57.52 mmol) and 4-(methoxymethoxy)-3-methyl- benzaldehyde (2487.51 mg, 13.8 mmol) in ethanol (120 mL) was stirred at 30 °C for 16 hours. After the reaction was completed, the mixture was adjusted to pH -4.0 with 1N HC1 solution. The resulting suspension was then filtered and solid was washed by water (200mL) and EtOH (100 mL) in sequence. The solid was concentrated in vacuo to give the crude product of (E)-l- (4-bromo-3-chloro-2-hydroxy-phenyl)-3-[4-(methoxymethoxy)-3-methyl-phenyl]prop-2-en-l- one (4.0 g, 43% yield) as a yellow solid. MS obsd. (ESL) [(M+H)+]: 411.1. Step 3: Preparation of 7-bromo-8-chloro-2-(4-hydroxy-3-methyl-phenyl)chromen-4-one
Figure imgf000133_0001
72c
To a solution of (£’)-l-(4-bromo-3-chloro-2-hydroxy-phenyl)-3-[4-(methoxymethoxy)-3- methyl-phenyl]prop-2-en-l-one (600.0 mg, 1.46 mmol) in DMSO (6 mL) was added iodine
(36.99 mg, 0.150 mmol) and the mixture was stirred at 140 °C for 4 hours. After the reaction was completed, the mixture was poured into water (60 mL). The resulting suspension was then filtered and solid was washed by Na2S03 solution (2 mol/L, lOmL) and water (50mL) in sequence. The solid was concentrated in vacuo to give the crude product of 7-bromo-8-chloro-2- (4-hydro xy-3-methyl-phenyl)chromen-4-one (430 mg, 44% yield,) as a yellow solid. MS obsd. (ESI+) [(M+H)+]: 365.0.
Step 4: Preparation of methyl 3-[2-[4-(7-bromo-8-chloro-4-oxo-chromen-2-yl)-2-methyl- phenoxy ] ethoxy] cyclobutanecarboxy late
Figure imgf000133_0002
72d
To a solution of 7-bromo-8-chloro-2-(4-hydroxy-3-methyl-phenyl)chromen-4-one (340.0 mg, 0.930 mmol, 1 eq), potassium carbonate (385.59 mg, 2.79 mmol, 3 eq) in DMF (lOmL) was added methyl 3-[2-(p-tolylsulfonyloxy)ethoxy]cyclobutanecarboxylate (335.92 mg, 1.02 mmol, 1.1 eq) and the mixture was stirred at 80 °C for 4 hours. After the reaction was complete, the mixture was diluted with water (50 mL) and the resulting mixture was extracted with EtOAc (30 mL) three times. The combined organic layer was washed with brine, dried over Na2S04 and concentrated in vacuo. The residue was purified by preparative HPLC to give two diastereomers of the methyl 3-[2-[4-(7-bromo-8-chloro-4-oxo-chromen-2-yl)-2-methyl- phenoxy] ethoxy] cyclobutanecarboxy late with cis- and Irons- configuration, one of which is appointed as 72d-A (82 mg, 14% yield, purity 100%) and the other is 72d-B (170 mg, 19% yield, purity 63.69%).MS obsd. (ESI+) [(M+H)+]: 520.9.
Step 5: Preparation of methyl 3-[2-[4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2-yl]-2- methyl-phenoxy]ethoxy]cyclobutanecarboxylate
Figure imgf000134_0001
72e To a solution of methyl 3-[2-[4-(7-bromo-8-chloro-4-oxo-chromen-2-yl)-2-methyl- phenoxy] ethoxy] cyclobutanecarboxy late (72d-A, 82.0 mg, 0.160 mmol) in DMF(5 mL) was added iodocopper (59.86 mg, 0.310 mmol), methyl 2,2-difluoro-2-(fluorosulfonyl)acetate
(150.95 mg, 0.790 mmol) and the mixture was then stirred at 120 °C for 6 hours. After the reaction was completed, the reaction was diluted with water (20 mL) and the resulting mixture was extracted with EtOAc (20 mL) three times. The combined organic layer was washed with brine, dried over Na2S04 and concentrated in vacuo to give the crude of methyl 3-[2-[4-[8- chloro-4-oxo-7-(trifluoromethyl)chromen-2-yl]-2-methyl- phenoxy] ethoxy] cyclobutanecarboxy late (62 mg, 48% yield, purity 33.91%) as a yellow solid, which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 511.1
Step 6: Preparation of 3-[2-[4-[8-chloro-4-oxo-7-(trifhioromethyl)chromen-2-yl]-2-inethyl- phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000135_0001
72
To a solution of methyl 3-[2-[4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2-yl]-2- methyl-phenoxy] ethoxy] cyclobutanecarboxylate (62.0 mg, 0.120 mmol) in the mixed solvent of THF (1 mL) and water (0.5 mL) was added lithium hydroxide (4.36 mg, 0.180 mmol) and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the mixture was adjusted to pH- 5 by addition of HC1 solution (2M). The resulting mixture was extracted with EtOAc (20mL) three times. The combined organic layer was concentrated in vacuo and the residue was purified by preparative HPLC to give 3-[2-[4-[8-chloro-4-oxo-7- (trifluoromethyl)chromen-2-yl]-2-methyl-phenoxy]ethoxy]cyclobutanecarboxylic acid (9.5 mg, 14% yield). lH NMR (DMSC )-d6, 400MHz): d ppm 12.18 (br s, 1H), 8.13 (d, 7=8.31 Hz, 1H), 7.81-8.06 (m, 3H), 7.09-7.21 (m, 2H), 4.13-4.27 (m, 3H), 3.70 (br s, 2H), 2.87-3.00 (m, 1H), 2.40 (ddd, 7=3.48, 6.82, 12.93 Hz, 2H), 2.26 (s, 3H), 2.09-2.19 (m, 2H). MS obsd. (ESI+)
[(M+H)+]: 497.0.
Example 73
C¾-3-[2-[2-chloro-4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2- yl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000135_0002
73
Step 1: Preparation of 2-benzyloxy ethyl trifluoromethanesulfonate
Figure imgf000136_0001
73a
To a solution of 2-benzyloxyethanol (2.0 g, 13.4 mmol) and 2,6-dimethylpyridine (2.8 g, 26.8 mmol) in dichloro methane ( 40 mL) was added trifluoromethanesulfonic anhydride (7.4 g, 26.8 mmol) at -30°C and the mixture was then stirred at 0 °C for 1 hour. The mixture was then washed with 1 N HC1 (20 mL) twice, water (20 mL) twice, brine (20 mL), dried over anhydrous Na2S04 and concentrated in vacuo to give the crude of 2-benzyloxyethyl
trifluoromethanesulfonate (4.0 g, 100% yield) , which was used in the next step directly without further purification. MS obsd. (ESI+) [(M+H)+]: 307.3.
Step 2: Preparation of cis-tert- butyl 3-(2-benzyloxyethoxy)cyclobutanecarboxylate
Figure imgf000136_0002
73b
To a solution of cis- /e/7-butyl 3-hydroxycyclobutanecarboxylate (CAS #: 939768-64-6, Cat.#: B253665, from BePharm Ltd, 2.26g, 18 mmol) in THF (20 mL) was added NaH (315 mg, 7.9 mmol) portion wise at 0 °C and the mixture was then stirred at 0 °C for 30 minutes. Then to the resulting mixture was added the solution of 2-benzyloxyethyl trifluoromethanesulfonate (4.0g, 13.4 mmol, crude) in THF (40 mL) dropwise at 0 °C. After addition, the mixture was poured into ice- water (50 mL) and extracted with dichloromethane (50 mL) three times. The combined organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo. The residue was purified by column chromatography on silica gel (elution with PE:EtOAc= 100: 1 to 3: 1) to give
Figure imgf000136_0003
/7-butyl 3-(2-benzyloxyethoxy)cyclobutanecarboxylate (1.5 g, 37.3% yield) as a yellow oil. MS obsd. (ESI+) [(M+Na)+]: 329.2.
Step 3: Preparation of cis-tert- butyl 3-(2-hydroxyethoxy)cyclobutanecarboxylate
Figure imgf000136_0004
73c Compound 73c was prepared in analogy to the procedure described for the preparation of Int-1 by using cis-tert- butyl 3-(2-benzyloxyethoxy)cyclobutanecarboxylate as the starting materials instead of methyl 3-(2-hydroxyethoxy)cyclobutanecarboxylate in Step 2.
Step 4: Preparation of cis-tert- butyl 3-[2-(p-tolylsulfonyloxy)ethoxy]cyclobutanecarboxylate
Figure imgf000137_0001
73d
Compound 73d was prepared in analogy to the procedure described for the preparation of intermediate Int-2 by using cis-tert- butyl 3-(2-hydroxyethoxy)cyclobutanecarboxylate as the starting materials instead of methyl 3-(2-hydroxyethoxy)cyclobutanecarboxylate.
Step 5: Preparation of 8-chloro-2-(3-chloro-4-hydroxy-phenyl)-7- (trifluoromethyl)chromen-4-one
Figure imgf000137_0002
Compound 73e was prepared in analogy to the procedure described for the preparation of compound 72c by using 3-chloro-4-hydroxy-benzaldehyde as the starting materials instead of 4- hydroxy-3-methyl-benzaldehyde in the step 1.
Step 6: Preparation of cis-tert- butyl 3-[2-[4-(7-bromo-8-chloro-4-oxo-chromen-2-yl)-2- chloro-phenoxy]ethoxy]cyclobutanecarboxylate
Figure imgf000137_0003
73f Compound 73f was prepared in analogy to the procedure described for the preparation of compound 72d by using 8-chloro-2-(3-chloro-4-hydroxy-phenyl)-7-(trifluoromethyl)chromen-4- one and cis-tert- butyl 3-[2-(p-tolylsulfonyloxy)ethoxy]cyclobutanecarboxylate as the starting material instead of 7-bromo-8-chloro-2-(4-hydroxy-3-methyl-phenyl)chromen-4-one and methyl 3-[2-(p-tolylsulfonyloxy)ethoxy]cyclobutanecarboxylate in the step 4.
Step 7: Preparation of cis-3-[2-[2-chloro-4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2- yl] phenoxy ] ethoxy] cyclobutanecarboxy lie acid
Figure imgf000138_0001
73
Example 73 was prepared in analogy to the procedure described for the preparation of example72 by using cis-tert- butyl 3-[2-[4-(7-bromo-8-chloro-4-oxo-chromen-2-yl)-2-chloro- phenoxy] ethoxy] cyclobutanecarboxy late as the starting materials instead of methyl 3-[2-[4-(7- bromo-8-chloro-4-oxo-chromen-2-yl)-2-methyl-phenoxy]ethoxy]cyclobutanecarboxylate in the step 5.
Example 73: NMR DMSO-d , 400MHz): d ppm 9.03 (s, 1H), 8.63 (d, 7=5.7 Hz, 1H), 7.98- 8.06 (m, 2H), 7.51 (t, 7=7.9 Hz, 1H), 7.34 (d, 7=5.8 Hz, 1H), 7.16 (s, 1H), 4.36-4.40 (s, 2H), 3.92-3.97 (m, 1H), 3.72-3.76 (m, 2H), 2.38-2.40 (m, 3H), 1.96-1.98 (m, 2H). MS obsd. (ESI+) [(M+H)+]:5l7.l.
BIOLOGICAL EXAMPLES
Example 74: Engineered HepDESl9 primary screen assay
The assay was employed to screen for novel cccDNA inhibitors. HepDESl9 is a cccDNA- producing cell line. In this cell line, HBeAg in the cell culture supernatant as surrogate marker, as HBeAg production depends on cccDNA level and activity. HepDESl9 is an engineered cell line which contains a 1.1 unit length HBV genome, and pgRNA transcription from the transgene is controlled by Tetracycline (Tet). In the absence of Tet, pgRNA transcription will be induced, but HBV e antigen (HBeAg) could not be produced from this pgRNA due to very short leader sequence before the HBeAg start codon and the start codon is disrupted. Only after cccDNA is formed, the missing leader sequence and start codon mutation would be restored from the 3’- terminal redundancy of pgRNA, and then HBeAg could be synthesized. Therefore, HBeAg could be used as a surrogate marker for cccDNA (Zhou, T. et al., Antiviral Res. (2006), 72(2), 116-124; Guo, H. et al., J. Virol. (2007), 81(22), 12472-12484).
HepDESl9 cells were seeded at 2xl06 cells per T150 flask and cultured with the culture medium (Dulbecco’s Modified Eagle Medium: Nutrient Mixture F-12 [DMEM-F12, Gibco Cat. 11320-82], 10% Fetal Bovine Serum [FBS, Clontech Cat. 631101], O.lmM Non-Essential Amino Acids Solution [NEAA, Gibco Cat. 11140-050], 50pg/mF Penicillin-Streptomycin [PS, Invitrogen Cat. 15140-163], 500 pg/mL Geneticin [G418, Invitrogen Cat. 10131-027]) containing 3pg/mP Tet (Sigma, Cat. 87128) for 5 days. Cells were then seeded at 4xl06 cells per T 150 in the same culture medium as described above in the absence of Tet for 8 days. Cells were then harvested and frozen at density of 2xl06 cells per mF. For compound testing, the frozen cells were thawed and seeded into 96- well plates at a density of 6 xlO4 cells per well. At 24hrs after seeding, half log serial dilutions of compounds made with Dimethyl sulfoxide (DMSO, Sigma, Cat. D2650) were further diluted with the same culture medium as described above before they were added to the cells to reach desired final compound concentrations and 1% DMSO concentration. Plates were then incubated at 37°C for another 5 days before measurement of HBeAg level and cell viability. Intracellular HBeAg level were measured with enzyme-linked immunosorbent assay (EFISA) kit (Shanghai Kehua Diagnostic Medical Products Co., Ftd). Cell viability was assessed using Cell Counting Kit-8 (Donjindo, Cat. CK04-20). IC50 values were derived from the dose-response curve using 4 parameter logistic curve fit method.
The compounds of the present invention were tested for their capacity to inhibit extracellular HBeAg level as described herein. The compounds of this invention were found to have IC50 below 50 mM. Particular compounds of formula (I) were found to have IC50 below 5.0 pM. Results of HepDESl9 primary screen assay are given in Table 1.
Table 1: Activity data in HepDESl9 primary screen assay
Figure imgf000139_0001
Figure imgf000140_0001
Example 75: cccDNA Southern Blot assay:
HepDESl9 cells were seeded at 4xl06 cells per T150 in the culture medium (Dulbecco’s Modified Eagle Medium: Nutrient Mixture F-12 [DMEM-F12, Gibco Cat. 11320-82], 10% Fetal Bovine Serum [FBS, Clontech Cat. 631101], O.lmM Non-Essential Amino Acids Solution [NEAA, Gibco Cat. 11140-050], 50pg/mF Penicillin-Streptomycin [PS, Invitrogen Cat. 15140- 163], 500 pg/iuL Geneticin [G418, Invitrogen Cat. 10131-027]) in the absence of Tet for 8 days. These cells were seeded at the density of lxlO6 cells per well in 6-well plate. At 24hrs after seeding, serial dilutions of compounds made with DMSO (Sigma, Cat. D2650) were further diluted with the same culture medium as described above before they were added to the cells to reach desired final compound concentrations and 1% DMSO concentration. After 5 days’ compound treatment, the cells growing in one well from 6-well plate were re-suspended with 500pL re-suspension buffer (50mM tris[hydroxymethyl] amino methane pH 7.5), lOmM ethylenediaminetetraacetic acid (EDTA), 50pg/mP RNase A [Qiagen, Cat. 158922] 500pL of 1.2% sodium dodecyl sulfate (SDS) was then added into the re-suspended cells to lyse the cells.
After 15 minutes’ incubation, 700mE precipitation buffer (3M cesium chloride, 1M potassium acetate, 0.67M acetic acid) was added and the lysate was incubated at 4°C for 2h. The lysate was centrifuged at 15000 revolutions per minute (RPM) at 4 °C for 15 minutes. The supernatant was collected and loaded onto Qiagen miniprep columns (QIAprep spin Mini prep kit, Cat. No.
27016). After centrifugation for 1 minute at 15000 RPM, the column was washed once with 750 pL wash buffer PE (QIAprep spin Mini prep kit, Cat. No. 27016). 80 pL of double distilled water was loaded to the columns to elute Hirt DNA.
Hirt DNA of each sample was loaded into 1.2% lx tris-acetate electrophoresis (TAE) agarose gel and separated at 90 voltages for 3 hours. The gel was then treated in 50mM NaAc- HAc, pH4.2 for 30min at room temperature (RT), and then denatured by soaking in denaturation buffer (0.5 M sodium hydroxide, 1.5 M sodium chloride) at RT for 30-45 min. The gel was then treated with neutralization buffer (1M tris[hydroxymethyl] amino methane pH7.4 and 1.5M NaCl) at RT for 30-45 min.
The gel was transferred onto a pre-wet Nylon membrane (GE life science, Hybond N+) by capillary transfer method overnight, followed by UV crosslinking. The membrane was transferred into a hybridization tube, then rinsed with double distilled water at 60 °C for 5 min.
10 mL of hybridization buffer (Lab kits, China) was added, the resulting sample was rotated in hybridization oven at 60 °C for 1 hour. Digoxigenin (DIG)-labelled HBV probe was denatured at 95 °C for 10 minutes, and then 7 pL of denatured probe was added to the hybridization tube, which was rotated in hybridization oven at 60 °C overnight.
On the second day, the membrane was washed according to the procedure of DIG wash and block buffer set kit (Roche, Cat. 11 585 762 001), and then incubated with 50mL antibody solution (Antibody anti-Digoxigenin-AP Fab fragment [Roche Cat. 11093274910] diluted in fresh lxblocking buffer at 1: 10,000) for 1 hour. The membrane was washed with 50 mL washing buffer (lxMaleic buffer with 0.3% Tween-20) for 15 minutes twice, and equilibrated with 20mL detection buffer (0.1M tris[hydroxymethyl] amino methane pH9.5, 0.1M sodium chloride) for 5 minutes. CDP-Star substrate (Roche, Cat. 12041677001) was added to the membrane for 5 minutes, and then the membrane was scanned by Bio-Rad Visualize Image System (Biorad, ChemiDoc-MP, Serial No. 731BR00916).
Results of cccDNA Southern Blot assay are given in Figure 1. The results indicate that the compounds of this invention dose-dependently reduced cccDNA level in HepDESl9 cells.
Example 76: Cryopreserved primary human hepatocytes (PHH) assay This assay is used to confirm the anti-HBV effect of the compounds in HBV PHH infection assay. Cryopreserved PHH (BioreclamationIVT, Lot YJM) was thawed at 37°C and gently transferred into pre-warmed InVitroGRO HT medium(BioreclamationIVT, Cat. S03317). The mixture was centrifuged at 70 relative centrifugal force (RCF) for 3 minutes at RT, and the supernatant was discarded. Pre-warmed InVitroGRO CP medium (BioreclamationIVT, Cat# S03316) was added to the cell pellet to gently re-suspend cells. The cells were seeded at the density of 5.8 x 104 cells per well to collagen I coated 96-well plate (Gibco, Cat. Al 142803) with the InVitroGRO CP medium. All plates were incubated at 37°C with 5% C02 and 85% humidity.
At 20 hours after plating, the medium was changed to PHH culture medium (Dulbecco's Modified Eagle Medium (DMEM)/Fl2 (1:1) (Gibco, Cat. 11320-033), 10% fetal bovine serum (Gibco Cat. 10099141), 100 U/mL penicillin, 100 pg/mL streptomycin (Gibco, Cat. 151401-122), 5 ng/mL human epidermal growth factor (Invitrogen Cat. PHG0311L), 20 ng/mL dexamethasone (Sigma, Cat. D4902) and 250 ng/mL human recombinant insulin (Gibco, Cat. 12585-014)). And the cells were incubated at 37°C with 5% C02 and 85% humidity for 4 hours. The medium was then changed to pre-warmed PHH culture medium containing 4% polyethylene glycol (PEG) MW8000 (Sigma, Cat. P1458-50ML) and 1% DMSO (Sigma, Cat. D2650). 5.8 x 106 genomic equivalents of HBV were added into the medium.
At 24 hours post-infection, the cells were gently washed with PBS and refreshed with PHH culture medium supplemented with 1% DMSO, and 0.25mg/mL Matrix gel (Corning, Cat.
356237) at 200pL per well. All plates were immediately placed in at 37°C C02 incubator.
24 hours later, serial dilutions of compounds made with DMSO were further diluted with the same culture medium (PHH culture medium supplemented with 1% DMSO and 0.25mg/mL Matrix gel as described above) before they were added to the cells to reach desired final compound concentrations and 1% DMSO concentration. The medium containing the compounds were refreshed every three days.
At 9 days post-compound treatment, extracellular HBsAg level were measured with
Chemiluminescence Immuno Assay (CLIA) kit (Autobio, HBsAg Quantitative CLIA).
Extracellular HBV DNA was extracted by MagNA Pure 96 system (Roche) and then determined by quantitative PCR with the following primers and probe:
HBV-Forward Primer (SEQ ID NO:l): AAG AAAAACCCCGCCTGT AA (5' to 3'); HBV-Re verse Primer (SEQ ID NO:2): CCTGTTCTGACTACTGCCTCTCC(5' to 3');
HBV-Probe: 5’+ tetramethylrhodamine + SEQ ID NO:3 + black hole quencher 2-3', wherein SEQ ID NOG is CCTG ATGTGATGTTCTCCATGTTCAGC .
HBsAg IC50 and HBV DNA IC50 values were derived from the dose-response curve using 4 parameter logistic curve fit method. The compounds of formula (I) have HBsAg IC50 <20 mM, particularly <1 mM; and HBV DNA IC50 < 50 pM. Test results of the compounds of this invention as well as the reference compounds in Cryopreserved PHH assay are given in Table 2 and Table 3.
Table 2: HBsAg IC50 data in Cryopreserved PHH assay
Figure imgf000143_0001
Table 3: HBV DNA IC50 in Cryopreserved PHH assay
Figure imgf000143_0002

Claims

1. A compound of the formula (I),
Figure imgf000144_0001
wherein
W is O or S;
Ai is CH or CR4;
A2 is CH or CR4;
A3 is CH or CR4;
A4 is N, CH or CR4;
As is N, CH or CR4;
A6 is CH;
A7 is N or CH;
R1 is halogen, Ci-6alkyl or C3-7cycloalkyl;
R2 is H, OH, halogen, Ci-6alkoxy, haloCi-6alkoxy or phenylCi-6alkoxy;
R3 is carboxy or Ci-6alkoxycarbonyl;
R4 is halogen, OH, CN, Ci-6alkyl, C3-7cycloalkyl, Ci-6alkoxy, oxopyrrolidinyl, morpholinyl or haloCi-6alkyl;
Gi is Ci-6alkyl, hydroxyCi-6alkyl or C3-7cycloalkylCi-6alkyl;
G2 is Ci-6alkyl, C3-7cyclo alkyl or phenyl;
or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
2. A compound according to claim 1, wherein
W is O or S;
Ai is CH or CR4;
A2 is CH or CR4;
A3 is CH or CR4; A4 is N, CH or CR4;
A5 is N, CH or CR4;
A6 is CH;
A7 is N or CH;
R1 is Cl, Br, methyl or cyclopropyl;
R2 is H, OH, Cl, Br, methoxy, trifluoromethoxy or benzyloxy;
R3 is carboxy, methoxycarbonyl or ethoxycarbonyl;
R4 is F, Cl, Br, OH, CN, methyl, isopropyl, cyclopropyl, methoxy, ethoxy, isopropoxy, oxopyrrolidinyl, morpholinyl or trifluoromethyl;
Gi is cyclobutylmethyl, ethyl, hydroxypropyl, methylethyl or propyl;
G2 is cyclobutyl, cyclohexyl, cyclopentyl, cyclopropyl, dimethylethyl, ethyl, methyl and phenyl;
or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
3. A compound according to claim 1 or 2, wherein Ai is CH.
4. A compound according to any one of claims 1 to 3, wherein A4 is CH or CR4.
5. A compound according to any one of claims 1 to 4, wherein As is CH or CR4.
6. A compound according to any one of claims 1 to 5, wherein A7 is CH.
7. A compound according to any one of claims 1 to 6, wherein R1 is halogen.
8. A compound according to any one of claims 1 to 7, wherein R2 is H.
9. A compound according to any one of claims 1 to 8, wherein R3 is carboxy.
10. A compound according to any one of claims 1 to 9, wherein R4 is halogen, Ci-6alkyl or Ci- 6alkoxy.
11. A compound according to any one of claims 1 to 10, wherein Gi is Ci-6alkyl.
12. A compound according to any one of claims 1 to 11, wherein G2 is Ci-6alkyl or C v 7cyclo alkyl.
13. A compound according to any one of claims 1 to 12, wherein
W is O or S;
Ai is CH;
A2 is CH or CR4;
A3 is CH or CR4;
A4 is CH or CR4;
A5 is CH or CR4;
A6 is CH;
A7 is CH;
R1 is halogen;
R2 is H;
R3 is carboxy;
R4 is halogen, Ci-6alkyl or Ci-6alkoxy;
Gi is Ci-6alkyl;
G2 is Ci-6alkyl or C3-7cycloalkyl.
14. A compound according to claim 13, wherein
W is O or S;
Ai is CH;
A2 is CH or CR4;
A3 is CH or CR4;
A4 is CH or CR4;
As is CH or CR4;
A6 is CH;
A7 is CH;
R1 is Cl, Br;
R2 is H;
R3 is carboxy;
R4 is F, Cl, Br, methyl, ethoxy or methoxy;
Gi is ethyl or methylethyl; 2 is cyclobutyl or methyl.
15. A compound according to claim 1 or 2, selected from
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
ethyl 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylate;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]benzoic acid;
ethyl 2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopropanecarboxylate;
cis-2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopropanecarboxylic acid; /mw5-2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopropanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclopentanecarboxylic acid;
cA-4-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclohexanecarboxylic acid;
/ra/7.v-4-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclohexanecarboxylic acid; 2-[3-[[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]methyl]cyclobutoxy]acetic acid;
2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]propanoic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy] -2,2-dimethyl-propanoic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-l-methyl-ethoxy]cyclobutanecarboxylic acid; 3-[3-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]propoxy]cyclobutanecarboxylic acid;
3-[3-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-2-hydroxy-propoxy]benzoic acid;
3-[2-[4-(8-chloro-7-fluoro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-7-methoxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-6-fluoro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(7,8-dichloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-6-cyano-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-7-cyano-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; cA-3-[2-[4-(8-chloro-5-hydroxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
cA-3-[2-[4-(8-chloro-5-methoxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
cA-3-[2-[4-(8-chloro-5-isopropoxy-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid; c 5,-3-[2-[4-(5-bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
r/.v-3- [2- [4-(5,8-dichloro-4-oxo-chromen-2-yl)phenoxy]ethoxy] cyclobutanecarboxylic acid; c/5,-3-[2-[4-(8-chloro-5-cyclopropyl-4-oxo-chromen-2- yl)phenoxy] ethoxy] cyclobutanecarboxylic acid;
r/.v-3-[2-[4-(8-chloro-5-methyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
r/.v-3- [2- [4-(8-bromo-4-oxo-chromen-2-yl)phenoxy]ethoxy] cyclobutanecarboxylic acid;
r/.v-3-[2-[4-(8-cyclopropyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; r/.v-3- [2- [4-(8-methyl-4-oxo-chromen-2-yl)phenoxy]ethoxy] cyclobutanecarboxylic acid;
2-[2-[4-(8-chloro-5-fluoro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetic acid;
r/.v-3-[2-[[5-(8-chloro-4-oxo-chromen-2-yl)-2-pyridyl]oxy]ethoxy]cyclobutanecarboxylic acid; / /Ύ//7 -3- [2- [[5-(8-chloiO-4-oxo-chromen-2-yl)-2-pyridyl]oxy]ethoxy] cyclobutanecarboxylic acid;
r/v-3-[2-[5-(8-chloiO-4-oxo-chiOirien-2-yl)pyrazin-2-yl]oxyethoxy]cyclobutanecarboxylic acid;
r/x- methyl 3-[2-[5-(8-chloro-4-oxo-chromen-2-yl)pyrazin-2- yl] oxyethoxy] cyclobutanecarboxylate;
r/x- methyl 3-[2-[6-(8-chloro-4-oxo-chromen-2-yl)pyridazin-3- yl] oxyethoxy] cyclobutanecarboxylate;
r/s-3-[2-[4-(8-chloiO-4-oxo-chromen-2-yl)-2-fluoiO-phenoxy]ethoxy]cyclobutanecarboxylic acid;
r/s-3-[2-[2-chloiO-4-(8-chloiO-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
r/s-3-[2-[2-biOmo-4-(8-chloiO-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
r/s-3-[2-[4-(8-chloiO-4-oxo-chiOmen-2-yl)-2-cyclopropyl- phenoxy] ethoxy] cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-cyano-phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-methyl-phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-isopropyl-phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-methoxy-phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-ethoxy-phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(3,8-dichloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
c/5,-3-[2-[4-(8-chloro-3-hydroxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
r/.v-3-[2-[4-(8-chloiO-3-methoxy-4-oxo-chi omen-2-yl)phenoxy]ethoxy]cyclobutanecai boxylic acid;
3-[2-[4-(8-chloro-4-oxo-thiochromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
2-[2-[4-(8-chloro-4-oxo-thiochromen-2-yl)phenoxy]ethoxy] acetic acid;
2-[2-[4-(8-chloro-3-hydroxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-morpholino-phenoxy]ethoxy]cyclobutanecarboxylic acid;
2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetamide;
Methyl 3-[2-[4-[8-chloro-6-fluoro-4-oxo-3-(trifluoromethoxy)chromen-2- yl]phenoxy] ethoxy] cyclobutanecarboxylate ;
3-[2-[4-[8-chloro-6-fluoro-4-oxo-3-(trifluoromethoxy)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(3-bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(3-benzyloxy-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-[8-chloro-4-oxo-6-(trifluoromethyl)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-6-cyclopropyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-[8-chloro-4-oxo-6-(2-oxopyrrolidin-l-yl)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-7-cyclopropyl-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
ethyl 2-[2-[4-(7-bromo-8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetate; 3-[2-[4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2-yl]-2-methyl- phenoxy] ethoxy] cyclobutanecarboxy lie acid; and
CA-3-[2-[2-chloro-4-[8-chloro-4-oxo-7-(trifluoromethyl)chromen-2- yl]phenoxy]ethoxy]cyclobutanecarboxylic acid;
or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
16. A compound according to claim 13 or 14, selected from:
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
r A-3- [2- [4-(8-chloiO-4-oxo-chromen-2-yl)phenoxy]ethoxy] cyclobutanecarboxy lie acid;
trans-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]-l-methyl-ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-6-fluoro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; cA-3-[2-[4-(8-chloro-5-methoxy-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
r A-3- [2- [4-(5,8-dichloiO-4-oxo-chromen-2-yl)phenoxy]ethoxy] cyclobutanecarboxy lie acid; r A-3- [2- [4-(8-biOiuo-4-oxo-chiOiuen-2-yl)phenoxy]ethoxy] cyclobutanecarboxy lie acid;
cA-3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-fluoro-phenoxy]ethoxy]cyclobutanecarboxylic acid;
cA-3-[2-[2-chloro-4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
cA-3-[2-[2-bromo-4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid;
3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-2-methyl-phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-chromen-2-yl)-3-ethoxy-phenoxy]ethoxy]cyclobutanecarboxylic acid; 3-[2-[4-(8-chloro-4-oxo-thiochromen-2-yl)phenoxy]ethoxy]cyclobutanecarboxylic acid; 2-[2- [4-(8-chloro-4-oxo-thiochromen-2-yl)phenoxy]ethoxy]acetic acid; and
2-[2-[4-(8-chloro-4-oxo-chromen-2-yl)phenoxy]ethoxy]acetamide;
or pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof.
17. A process for the preparation of a compound according to any one of claims 1 to 16 comprising
(a) substitution of a compound of formula (X),
Figure imgf000151_0001
(X),
with a compound of formula (XI) in the presence of a base;
(b) substitution of a compound of formula (IX),
Figure imgf000151_0002
(ix),
with a compound of formula (XII) in the presence of a base;
(c) cyclization of compound of formula (XIV),
Figure imgf000151_0003
in the presence of a suitable Lewis acid;
wherein R1, R2, R3,W, Ai to A7, Gi and G2 are defined as any one of claims 1 to 14.
18. A compound according to any one of claims 1 to 16 for use as therapeutically active substance.
19. A pharmaceutical composition comprising a compound in accordance with any one of claims 1 to 16 and a therapeutically inert carrier.
20. The use of a compound according to any one of claims 1 to 16 for the treatment or prophylaxis of HBV infection.
21. The use of a compound according to any one of claims 1 to 16 for the preparation of a medicament for the treatment or prophylaxis of HBV infection.
22. The use of a compound according to any one of claims 1 to 16 for the inhibition of cccDNA.
23. The use of a compound according to any one of claims 1 to 16 for the inhibition of HBeAg.
24. The use of a compound according to any one of claims 1 to 16 for the inhibition of HBsAg.
25. The use of a compound according to any one of claims 1 to 16 for the inhibition of HBV DNA.
26. A compound according to any one of claims 1 to 16 for the treatment or prophylaxis of
HBV infection.
27. A compound according to any one of claims 1 to 16, when manufactured according to a process of claim 17.
28. A method for the treatment or prophylaxis of HBV infection, which method comprises administering an effective amount of a compound as defined in any one of claims 1 to 16.
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JP2023527281A (en) * 2020-05-18 2023-06-28 ソーク インスティテュート フォー バイオロジカル スタディーズ Chromen-4-one derivatives, such as flavones, for use as CK2 inhibitors for the treatment of neuroinflammation

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