Patent Grant
11566018
The present disclosure relates generally to the treatment of infectious disease.
Annually, Respiratory Syncytial Virus (RSV) causes an estimated 3.4 million severe lower respiratory infections requiring hospitalization in children under 5 years of agwe. Recent vaccine development efforts have not been fruitful, and no licensed efficacious therapeutics are available to treat infection. A viral RNA-dependent RNA polymerase complex (RdRp), required for expression and replication of the viral genome, provides an attractive target for inhibition of the viral replication cycle. The absence of an X-ray crystal structure of this complex rules out in silico approaches for inhibitor development.
ZIKV infection, in contrast to RSV, has only recently been identified as an unmet therapeutic need. The most recent severe ZIKV outbreak to date occurred in Brazil, with an estimated incidence of 30,000 infections since first detection in May 2015. While primary symptoms are usually mild, the association of ZIKV infection with congenital microcephaly and its mosquito-based transmission underscore the importance of developing therapeutics against it, especially from a prophylactic vantage point. Currently no vaccine or specific antiviral treatments are available for ZIKV.
In an aspect of the present application there is provided a compound of formula (I)
or
In another aspect, there is provided a compound having the formula (II)
or
In another aspect, there is provided a compound having the formula (III)
or
wherein:
Y and Y′ are each independently C or a heteroatom;
R1 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted;
R5 is independently absent, H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R5, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and
R6 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R6, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and wherein one Y′ is bonded to C1 and the corresponding R6 is absent.
In another aspect, there is provided a compound having the formula (IV)
or
R5 is independently absent, H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R5, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and
R7 is independently absent, H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R7, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted.
In another aspect, there is provided a compound having the formula (V)
or
R5 is independently absent, H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R5, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted;
R7 is independently absent, H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R7, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and,
R8 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R8, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted.
In another aspect, there is provided a compound having the formula (VI)
or
In an embodiment of the present application, there is provided a compound having the structure
In another embodiment, there is provided a compound having the structure
In another aspect of the present application, there is provided a method of synthesizing a compound as described herein, comprising:
In an embodiment of the present application, there is provided a method wherein step d) further comprises reacting the electrophilic C-acylimine with a nucleophilic compound in the presence of a Bronsted acid catalyst.
In another embodiment, there is provided a method wherein the transition metal catalyst is a
Cu catalyst. In another embodiment, the transition metal catalyst is Cu(hfacac)2, Cu(OTf)2, or CuOTf.Ph(CH3).
In another embodiment, there is provided a method wherein the nucleophilic compound is a heteroatom-containing compound. In another embodiment, the heteroatom-containing compound is a heterocycle or a cycle substituted with a heteroatom-containing moiety. In another embodiment, the heterocycle is pyrrole, furan, thiophene, pyridine, indole, benzofuran, benzothiphene, imidazole, or derivatives thereof, each of which is optionally substituted.
In another aspect of the present application, there is provided a pharmaceutical composition comprising a compound as described herein, or a compound syntheized by the method as described herein, and a pharmaceutically acceptable carrier, diluent, or vehicle.
In another aspect of the present application, there is provided a method of treating a subject having or suspected of having an infectious disease, comprising: administering a therapeutically effective amount of a compound as described herein, or a compound syntheized by the method as described herein, or a pharmaceutical composition as described herein.
In another aspect of the present application, there is provided a method of treating a subject having or suspected of having an infectious disease, comprising: administering a therapeutically effective amount of a compound as described herein, or a compound syntheized by the method as described herein, or a pharmaceutical composition as described herein, wherein said infectious disease is caused by a virus.
In another embodiment of the present application, there is provided a method wherein said virus is a virus from the family Flaviviridae. In another embodiment, the virus is from the genera Hepacivirus, Flavivirus, Pegivirus, or Pestivirus. In another embodiment, Flavivirus is yellow fever virus (YFV), Japanese encephalitis virus (JEV), Tick-borne encephalitis virus (TBEV), Dengue virus (DENV), West Nile virus (WNV), Zika virus (ZIKAV), or any combination thereof.
In another embodiment, there is provided a method wherein said virus is from the family Paramyxoviridae. In another embodiment, said virus is from the genera Paramyxovirus, Pneumovirus, or Morbillivirus. In another embodiment, Paramyxovirus is parainfluenza virus or mumpus virus. In another embodiment, Pneumovirus is respiratory syncytial virus (RSV). In another embodiment, said Morbillivirus is measles virus.
In another embodiment, there is provided a method wherein said subject is a human, a domesticated animal, livestock, a laboratory animal, a non-human mammal, a non-human primate, a rodent, a bird, a reptile, an amphibian, or a fish. In some cases, the individual to be treated with a method of the present disclosure is a human. In some cases, the individual to be treated with a method of the present disclosure is an ungulate (e.g., a bovine; an ovine; a caprine; an equine; etc.).
In another aspect of the present application, there is provided a use of a compound of as described herein, or a compound syntheized by the method as described herein, or a pharmaceutical composition as described herein for treating a subject having or suspected of having an infectious disease.
In another aspect of the present application, there is provided a use of a compound as described herein, or a compound syntheized by the method as described herein, or a pharmaceutical composition as described herein in the manufacture of a medicament for treating a subject having or suspected of having an infectious disease.
In another aspect of the present application, there is provided a use of a compound as described herein, or a compound syntheized by the method as described herein, or a pharmaceutical composition as described herein for treating a subject having or suspected of having an infectious disease, wherein said infectious disease is caused by a virus.
In another aspect of the present application, there is provided a use of a compound as described herein, or a compound syntheized by the method as described herein, or a pharmaceutical composition as described herein in the manufacture of a medicament for treating a subject having or suspect of having an infectious disease, wherein said infectious disease is caused by a virus.
In another embodiment of the present application, there is provided a use wherein said virus is a virus from the family Flaviviridae. In another embodiment, the virus is from the genera Hepacivirus, Flavivirus, Pegivirus, or Pestivirus. In another embodiment, Flavivirus is yellow fever virus (YFV), Japanese encephalitis virus (JEV), Tick-borne encephalitis virus (TBEV), Dengue virus (DENV), West Nile virus (WNV), Zika virus (ZIKAV), or any combination thereof.
In another embodiment of the present application, there is provided a use wherein said virus is a virus from the family Paramyxoviridae. In another embodiment, said virus is from the genera Paramyxovirus, Pneumovirus, or Morbillivirus. In another embodiment, Paramyxovirus is parainfluenza virus or mumpus virus. In another embodiment, Pneumovirus is respiratory syncytial virus (RSV). In another embodiment, said Morbillivirus is measles virus.
In another embodiment of the present application, there is provided a usewherein said subject is a human, a domesticated animal, livestock, a laboratory animal, a non-human mammal, a non-human primate, a rodent, a bird, a reptile, an amphibian, or a fish.
Embodiments of the present disclosure will now be described, by way of example only, with reference to the attached Figures.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
As used in the specification and claims, the singular forms “a”, “an” and “the” include plural references unless the context clearly dictates otherwise.
The term “comprising” as used herein will be understood to mean that the list following is non-exhaustive and may or may not include any other additional suitable items, for example one or more further feature(s), component(s) and/or ingredient(s) as appropriate.
As used herein, the term ‘optionally substituted’ refers to being substituted or unsubstituted.
As used herein, the term “unsubstituted” refers to any open valence of an atom being occupied by hydrogen. Also, if an occupant of an open valence position on an atom is not specified then it is hydrogen.
As used herein, the term “substituted” refers to having one or more substituents or substituent moieties whose presence either facilitates or improves a desired reaction/property, or does not impede a desired reaction/property. A “substituent” is an atom or group of bonded atoms that can be considered to have replaced one or more hydrogen atoms attached to a parent molecular entity; and, whose presence either facilitates or improves desired reactions, properties, and/or functions of an invention, or does not impede desired reactions, properties, and/or functions of an invention. Examples of substituents include alkyl, alkenyl, alkynyl, aryl, polycyclic aryl, benzyl, polycyclic benzyl, fused aromatic rings, arylhalide, heteroaryl, polycyclic heteroaryl, fused heteroaromatic rings, cycloalkyl (non-aromatic ring), halo, alkoxyl, perfluoronated alkoxyl, amino, alkylamino, alkenylamino, amide, amidine, hydroxyl, thioether, alkylcarbonyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carbonate, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate, phosphate ester, phosphonato, phosphinato, cyano, acylamino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, dithiocarboxylate, sulfate, sulfato, sulfonate, sulfamoyl, sulfonamide, Si(alkyl)3, Si(alkoxy)3, nitro, nitrile, azido, heterocyclyl, ether, ester, silicon-containing moieties, thioester, or a combination thereof. The substituents may themselves be substituted. For instance, an amino substituent may itself be mono or independently disubstituted by further substituents provided above, such as alkyl, alkenyl, alkynyl, aryl, aryl-halide, heteroaryl, cycloalkyl (non-aromatic ring).
As used herein, “alkyl” refers to a linear or branched saturated hydrocarbon moiety that consists solely of single-bonded carbon and hydrogen atoms, which can be unsubstituted or substituted with one or more substituents. Examples of saturated straight or branched chain alkyl groups include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-butyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl and 2-ethyl-1-butyl, 1-heptyl, and 1-octyl.
As used herein, “alkenyl” refers to a linear or branched hydrocarbon moiety that comprises at least one carbon to carbon double bond, which can be unsubstituted or substituted with one or more substituents. “Alkynyl” refers to a linear or branched hydrocarbon moiety that comprises at least one carbon to carbon triple bond, which can be unsubstituted or substituted with one or more substituents.
The term “carbocycle” as used herein refers to a non-aromatic, saturated or partially saturated monocyclic or polycyclic hydrocarbon ring moiety containing at least 3 carbon atoms. Examples of C3-Cn carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, norbornyl, adamantyl, bicyclo[2.2.2]oct-2-enyl, and bicyclo[2.2.2]octyl.
As used herein, “aryl” and/or “aromatic ring” refers to an aromatic (unsaturated cyclic) hydrocarbon moiety having 6 to 100 atoms, or 6 to 50 atoms, or 6 to 25 atoms, or 6 to 15 atoms, which can be unsubstituted or substituted with one or more substituents. The aromatic hydrocarbon moiety may be derived from benzene or a benzene derivative; may be monocyclic or polycyclic, where polycyclic may include a fused ring system. Examples include, but are not limited to, phenyl, naphthyl, xylene, phenyl ethane, substituted phenyl, substituted naphthyl, substituted xylene, substituted 4-ethylphenyl, benzyl, etc.
As used herein, “cycle” refers to an aromatic or nonaromatic monocyclic, polycyclic, or fused ring hydrocarbon moiety, which can be substituted or unsubstituted. Included within the term “cycle” are carbocycles and aryls, as defined above.
As used herein, “heteroaryl” or “heteroaromatic” refers to an aryl (including fused aryl rings) that includes heteroatoms selected from oxygen, nitrogen, sulfur, and phosphorus. A “heteroatom” refers to an atom that is not carbon or hydrogen, such as nitrogen, oxygen, sulfur, or phosphorus. Heteroaryl or heteroaromatic groups include, for example, furanyl, thiophenyl, pyrrolyl, imidazoyl, benzamidazoyl, 1,2- or 1,3-oxazolyl, 1,2- or 1,3-diazolyl, 1,2,3- or 1,2,4-triazolyl, and the like.
As used herein, a “heterocycle” is an aromatic or nonaromatic monocyclic, polycyclic, or fused ring moiety of carbon atoms and at least one heteroatom, or 1 to 4 heteroatoms, or 1 to 10 heteroatoms. A “heteroatom” refers to an atom that is not carbon or hydrogen, such as nitrogen, oxygen, sulfur, or phosphorus. Included within the term “heterocycle” is “heteroaryl”, which refers to an aromatic (unsaturated cyclic) moiety of carbon atoms and at least one heteroatom, or 1 to 4 heteroatoms, or 1 to 10 heteroatoms, having a total of 6 to 100 atoms, or 6 to 50 atoms, or 6 to 25 atoms, or 6 to 15 atoms, which can be unsubstituted or substituted with one or more substituents. Also included within this term are monocyclic and bicyclic rings that include one or more double and/or triple bonds within the ring. Examples of 3- to 9-membered heterocycles include, but are not limited to, furanyl, thiophenyl, pyrrolyl, imidazoyl, benzamidazoyl, 1,2- or 1,3-oxazolyl, 1,2- or 1,3-diazolyl, 1,2,3- or 1,2,4-triazolyl, aziridinyl, oxiranyl, thiiranyl, azirinyl, diaziridinyl, diazirinyl, oxaziridinyl, azetidinyl, azetidinonyl, oxetanyl, thietanyl, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, oxazinyl, thiazinyl, diazinyl, triazinyl, tetrazinyl, imidazolyl, benzimidazolyl, tetrazolyl, indolyl, isoquinolinyl, quinolinyl, quinazolinyl, pyrrolidinyl, purinyl, isoxazolyl, benzisoxazolyl, furanyl, furazanyl, pyridinyl, oxazolyl, benzoxazolyl, thiazolyl, benzthiazolyl, thiophenyl, pyrazolyl, triazolyl, benzodiazolyl, benzotriazolyl, pyrimidinyl, isoindolyl and indazolyl.
As used herein, “halo” refers to F, Cl, Br, I.
The term “subject”, as used herein, refers to an animal, and can include, for example, domesticated animals, such as cats, dogs, etc., livestock (e.g., cattle, horses, pigs, sheep, goats, etc.), laboratory animals (e.g., mouse, rabbit, rat, guinea pig, etc.), mammals, non-human mammals, primates, non-human primates, rodents, birds, reptiles, amphibians, fish, and any other animal. In a specific example, the subject is a human.
The term “treatment”, “treat”, or “treating” as used herein, refers to obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission (whether partial or total), whether detectable or undetectable. “Treating” and “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment.
The term “amelioration” or “ameliorates” as used herein refers to a decrease, reduction or elimination of a condition, disease, disorder, or phenotype, including an abnormality or symptom.
The term “functional derivative” as used herein refers to a molecule that retains a biological activity (either function or structural) that is substantially similar to that of the original compound. A functional derivative or equivalent may be a natural derivative or is prepared synthetically.
Also encompassed is prodrug or “physiologically functional derivative”. The term “physiologically functional derivative” as used herein refers to compounds which are not pharmaceutically active themselves but which are transformed into their pharmaceutically active form in vivo, i.e. in the subject to which the compound is administered. The term “prodrug” as used herein, refers to a derivative of a substance that, following administration, is metabolized in vivo, e.g. by hydrolysis or by processing through an enzyme, into an active metabolite.
In some cases, a compound of the present disclosure inhibits enzymatic activity of an RNA polymerase (e.g., an RNA-dependent RNA polymerase) encoded by an RNA virus. For example, in some cases, a compound of the present disclosure inhibits enzymatic activity of an RNA polymerase by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, compared to the enzymatic activity of the RNA polymerase in the absence of the compound.
An RNA polymerase (e.g., an RNA-dependent RNA polymerase) that can be inhibited by a compound of the present disclosure can be an RNA polymerase encoded by any of the following RNA viruses: a Coronaviridae virus, a Picornaviridae virus, a Caliciviridae virus, a Flaviviridae virus, a Togaviridae virus, a Bornaviridae, a Filoviridae, a Paramyxoviridae, a Pneumoviridae, a Rhabdoviridae, an Arenaviridae, a Bunyaviridae, an Orthomyxoviridae, a Deltavirus, Coronavirus, SARS coronavirus, MERS coronavirus, Poliovirus, Rhinovirus, Hepatitis A virus, Hepatitis B virus, Norwalk virus, Yellow fever virus, West Nile virus, Hepatitis C virus, Dengue fever virus, Zika virus, Rubella virus, Ross River virus, Sindbis virus, Chikungunya virus, Borna disease virus, Ebola virus, Marburg virus, Measles virus, Mumps virus, Nipah virus, Hendra virus, Newcastle disease virus, Human respiratory syncytial virus, Rabies virus, Lassa virus, Hantavirus, Crimean-Congo hemorrhagic fever virus, Influenza virus (e.g., swine influenza virus; avian influenza virus; H1N1, H3N2, H7N9, H5N1, etc.), porcine respiratory and reproductive disease syndrome virus (PRRSV), Seneca valley virus, porcine epidemic diarrhea virus (PEDV), porcine delta coronavirus (PDCV), porcine circoviral associated diseases (PCVAD), and Hepatitis D virus. In some cases, the RNA polymerase that is inhibited by a compound of the present disclosure is encoded by Zika virus. In some cases, the RNA polymerase that is inhibited by a compound of the present disclosure is encoded by Ebola virus. In some cases, the RNA polymerase that is inhibited by a compound of the present disclosure is encoded by human respiratory syncytial virus (RSV). In some cases, the RNA polymerase that is inhibited by a compound of the present disclosure is encoded by bovine RSV.
RNA-dependent RNA polymerases (RdRP) are known in the art. In some cases, an RNA polymerase (e.g., an RNA-dependent RNA polymerase) that is inhibited by a compound of the present disclosure comprises an amino acid sequence having at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, with the amino acid sequence depicted in
The present disclosure provides methods of treating a viral infection, the methods comprising administering to an individual having the viral infection a therapeutically effective amount of a compound of the present disclosure, or a composition comprising a compound of the present disclosure.
As used herein, the term “therapeutically effective amount” refers to an amount that is effective for preventing, ameliorating, or treating a disease or disorder (e.g., an infection disease, such as a viral disease).
In some cases, a therapeutically effective amount of a compound of the present disclosure is an amount that is effective to reduce the amount of virus in a tissue, organ, or fluid in an individual being treated. For example, in some cases, a therapeutically effective amount of a compound of the present disclosure is an amount that is effective to reduce the amount of virus in a tissue, organ, or fluid in an individual being treated by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or more than 80%, compared to the amount of virus present in the tissue, organ, or fluid in the individual before treatment with the compound.
In an aspect, there is described a compound, compositions, methods, and uses, for the treatment of a subject having, or suspected of having, an infectious disease.
In an example the infectious disease is caused by a virus.
Viral infections that can be treated with a method of the present disclosure include infections caused by any of the following: a Coronaviridae virus, a Picornaviridae virus, a Caliciviridae virus, a Flaviviridae virus, a Togaviridae virus, a Bornaviridae, a Filoviridae, a Paramyxoviridae, a Pneumoviridae, a Rhabdoviridae, an Arenaviridae, a Bunyaviridae, an Orthomyxoviridae, a Deltavirus, Coronavirus, SARS, Poliovirus, Rhinovirus, Hepatitis A virus, Hepatitis B virus, Norwalk virus, Yellow fever virus, West Nile virus, Hepatitis C virus, Dengue fever virus, Zika virus, Rubella virus, Ross River virus, Sindbis virus, Chikungunya virus, Borna disease virus, Ebola virus, Marburg virus, Measles virus, Mumps virus, Nipah virus, Hendra virus, Newcastle disease virus, Human respiratory syncytial virus, bovine respiratory syncytial virus, Rabies virus, Lassa virus, Hantavirus, Crimean-Congo hemorrhagic fever virus, Influenza virus (e.g., swine influenza virus; avian influenza virus; etc.), porcine respiratory and reproductive disease syndrome virus (PRRSV), Seneca valley virus, porcine epidemic diarrhea virus (PEDV), porcine delta coronavirus (PDCV), porcine circoviral associated diseases (PCVAD), and Hepatitis D virus. In some cases, the viral infection is caused by a positive-strand RNA virus. In some cases, the viral infection is caused by a negative-strand RNA virus.
In an example, the virus is from the family Flaviviridae. In another example, the virus I from the genera Hepacivirus, Flavivirus, Pegivirus, or Pestivirus
Non-limiting examples of flavivirus include yellow fever virus (YFV), Japanese encephalitis virus (JEV), Tick-borne encephalitis virus (TBEV), Dengue virus (DENV), West Nile virus (WNV), Zika virus (ZIKAV), or any combination thereof.
In a specific example, the viral infection is Zika virus.
In an example, the virus is from the family of Paramyxoviridae. In another example, the virus is from the genera Paramyxovirus, Pneumovirus, or Morbillivirus
Non-limiting example of paramyxovirus include parainfluenza virus and mumpus virus.
Non-limiting examples of Pneumovirus include respiratory syncytial virus (RSV).
Non-limiting examples of Morbillivirus include measles virus.
In a specific example, the viral infection is respiratory syncytial virus (RSV). As one example, in some cases, the viral infection is human RSV. As another example, in some cases, the viral infection is bovine RSV.
In another aspect, there is described a compound of formula (I)
or
a stereoisomer, a racemate, a tautomer, a pharmaceutically acceptable salt, a solvate, or a functional derivative thereof,
wherein:
R1 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted;
R2 is independently aryl, benzyl, or heterocycle, each of which is optionally substituted; and
R3 and R4 are each independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or R3 and R4, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted.
In an example, there is described a compound having the formula (II)
or
a stereoisomer, a racemate, a tautomer, a pharmaceutically acceptable salt, a solvate, or a functional derivative thereof,
wherein:
Y is independently C or a heteroatom;
R1 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted;
R3 and R4 are each independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or R3 and R4, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and
R6 is independently absent, H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R6, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and wherein one Y is bonded to C1 and the corresponding R6 is absent.
In another example, there is described a compound having the formula (III)
or
a stereoisomer, a racemate, a tautomer, a pharmaceutically acceptable salt, a solvate, or a functional derivative thereof,
wherein:
Y and Y′ are each independently C or a heteroatom;
R1 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted;
R5 is independently absent, H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R5, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and
R6 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R6, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and wherein one Y′ is bonded to C1 and the corresponding R6 is absent.
In another example, there is described a compound having the formula (IV)
or
a stereoisomer, a racemate, a tautomer, a pharmaceutically acceptable salt, a solvate, or a functional derivative thereof,
wherein:
Y is independently C or a heteroatom;
R1 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted;
R5 is independently absent, H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R5, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and
R7 is independently absent, H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R7, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted.
In another example, there is described a compound having the formula (V)
or
a stereoisomer, a racemate, a tautomer, a pharmaceutically acceptable salt, a solvate, or a functional derivative thereof,
wherein:
Y is independently C or a heteroatom;
R1 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted;
R5 is independently absent, H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R5, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted;
R7 is independently absent, H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R7, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and,
R8 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R8, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted.
In another example, there is described a compound having the formula (VI)
or
a stereoisomer, a racemate, a tautomer, a pharmaceutically acceptable salt, a solvate, or a functional derivative thereof,
wherein:
R1 is an ester;
R5 is a halo; and,
R8 is independently H, C1-C10 alkoxy, or halo, each of which is optionally substituted.
In some examples, the compounds as described herein have the structure
In some examples, the compounds as described herein have the structure
In other examples, the compounds as described herein have any one of the structure
In other examples, the compounds as described herein have any one of the structure
In other examples, the compounds as described herein have any one of the structure
In other examples, the compounds as described herein have any one of the structure
In other examples, the compounds as described herein have any one of the structure
In other examples, the compounds as described herein have any one of the structure
In other examples, the compounds as described herein have any one of the structure
In other examples, the compounds as described herein have any one of the structure
In other examples, the compounds as described herein have any one of the structure
In other examples, the compounds as described herein have any one of the structure
In another example, the absolute stereochemistry of the quaternary stereocentre (chiral carbon) of the compounds as described herein may be R, S, or a mixture of R and S.
In another example, when the compounds as described herein comprise two or more stereocentres (chiral carbons), the absolute stereochemistry may be any permutation of R and S, or a mixture of any permutation.
In another aspect, there is described a method of synthesizing the compounds as described herein, the method comprising:
In an example, of the method as described herein step d) further comprises reacting the electrophilic C-acylimine with a nucleophilic compound in the presence of a Bronsted acid catalyst.
In another example of the method as described herein, the transition metal catalyst is a Cu catalyst. In another example, the transition metal catalyst is Cu(hfacac)2, Cu(OTf)2, or CuOTf.Ph(CH3).
In another example of the method as described herein, the nucleophilic compound is a heteroatom-containing compound. In another example the heteroatom-containing compound is a heterocycle or a cycle substituted with a heteroatom-containing moiety. In another example, the heterocycle is pyrrole, furan, thiophene, pyridine, indole, benzofuran, benzothiphene, imidazole, or derivatives thereof, each of which is optionally substituted.
In some examples, the compounds as described herein are an enantiomer, a racemate, a tautomer, or a pharmaceutically acceptable salt, or a solvate, or a functional derivative thereof.
In some examples, there is described a composition comprising a compound as described herein, and a pharmaceutically acceptable carrier, diluent, or vehicle.
A compound or composition may be administered alone or in combination with other treatments, either simultaneously or sequentially, dependent upon the condition to be treated.
In treating a subject, a therapeutically effective amount may be administered to the subject.
Formulations may conveniently be presented in unit dosage form and may be prepared by any methods known in the art. Such methods include the step of bringing the active compound into association with a carrier, which may constitute one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active compound with liquid carriers or finely divided solid carriers or both, and then if necessary shaping the product.
The compounds and compositions may be administered to a subject by any convenient route of administration, whether systemically/peripherally or at the site of desired action, including but not limited to, oral (e.g. by ingestion); topical (including e.g. transdermal, intranasal, ocular, buccal, and sublingual); pulmonary (e.g. by inhalation or insufflation therapy using, e.g. an aerosol, e.g. through mouth or nose); rectal; vaginal; parenteral, for example, by injection, including subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, subarachnoid, and intrasternal; by implant of a depot, for example, subcutaneously or intramuscularly.
Compounds and/or compositions comprising compounds disclosed herein may be used in the methods described herein in combination with standard treatment regimes, as would be known to the skilled worker.
Methods of the invention are conveniently practiced by providing the compounds and/or compositions used in such method in the form of a kit. Such kit preferably contains the composition. Such a kit preferably contains instructions for the use thereof.
Examples of Non-Limiting Aspects of the Disclosure
Aspects, including embodiments, of the present subject matter described above may be beneficial alone or in combination, with one or more other aspects or embodiments. Without limiting the foregoing description, certain non-limiting aspects of the disclosure numbered 1-41 are provided below. As will be apparent to those of skill in the art upon reading this disclosure, each of the individually numbered aspects may be used or combined with any of the preceding or following individually numbered aspects. This is intended to provide support for all such combinations of aspects and is not limited to combinations of aspects explicitly provided below:
Aspect 1. A compound of formula (I)
or
a stereoisomer, a racemate, a tautomer, a pharmaceutically acceptable salt, a solvate, or a functional derivative thereof,
wherein:
R1 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, C1-C10 alkoxy, C10-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted;
R2 is independently aryl, benzyl, or heterocycle, each of which is optionally substituted; and R3 and R4 are each independently H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or R3 and R4, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted.
Aspect 2. The compound of aspect 1, having the formula (II)
or
wherein:
Y is independently C or a heteroatom;
R1 is independently H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted;
R3 and R4 are each independently H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or R3 and R4, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and
R6 is independently absent, H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R6, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and wherein one Y is bonded to C1 and the corresponding R6 is absent.
Aspect 3. The compound of aspect 1 or 2, having the formula (III)
or
a stereoisomer, a racemate, a tautomer, a pharmaceutically acceptable salt, a solvate, or a functional derivative thereof,
wherein:
Y and Y′ are each independently C or a heteroatom;
R1 is independently H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted;
R5 is independently absent, H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R5, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and
R6 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R6, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and wherein one Y′ is bonded to C1 and the corresponding R6 is absent.
Aspect 4. The compound of any one of aspects 1 to 3, having the formula (IV)
or
a stereoisomer, a racemate, a tautomer, a pharmaceutically acceptable salt, a solvate, or a functional derivative thereof,
wherein:
Y is independently C or a heteroatom;
R1 is independently H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted;
R5 is independently absent, H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R5, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and
R7 is independently absent, H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R7, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted.
Aspect 5. The compound of any one of aspects 1 to 4, having the formula (V)
or
a stereoisomer, a racemate, a tautomer, a pharmaceutically acceptable salt, a solvate, or a functional derivative thereof,
wherein:
Y is independently C or a heteroatom;
R1 is independently H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted;
R5 is independently absent, H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R5, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted;
R7 is independently absent, H, C1-C10 alkyl, C10-C20 alkyl, C2-C10 alkenyl, C10-C20 alkenyl, C2-C10 alkynyl, C2-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R7, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted; and,
R8 is independently H, C1-C10 alkyl, C2-C20 alkyl, C2-C10 alkenyl, C2-C20 alkenyl, C2-C10 alkynyl, C10-C20 alkynyl, C3-C20 carbocycle, aryl, benzyl, heterocycle, C1-C10 alkoxy, C2-C20 alkoxy, alcohol, ether, ketone, carboxylic acid, ester, thiol, thioether, amine, amide, carbamate, nitro, cyano, or halo, each of which is optionally substituted; or two of R8, together with the atoms to which they are attached, are connected to form a cycle or heterocycle, each of which is optionally substituted.
Aspect 6. The compound of any one of aspects 1 to 5, having the formula (VI)
or
a stereoisomer, a racemate, a tautomer, a pharmaceutically acceptable salt, a solvate, or a functional derivative thereof,
wherein:
R1 is an ester;
R5 is a halo; and,
R8 is independently H, C1-C10 alkoxy, or halo, each of which is optionally substituted.
Aspect 7. The compound of any one of aspects 1 to 6, having the structure
Aspect 8. The compound of any one of aspects 1 to 4, having the structure
Aspect 9. A method of synthesizing a compound of any one of aspects 1 to 8, comprising:
Aspect 10. The method of aspect 9, wherein step d) further comprises reacting the electrophilic C-acylimine with a nucleophilic compound in the presence of a Bronsted acid catalyst.
Aspect 11. The method of aspect 9 or 10, wherein the transition metal catalyst is a Cu catalyst.
Aspect 12. The method of aspect 11, wherein the transition metal catalyst is Cu(hfacac)2, Cu(OTf)2, or CuOTf.Ph(CH3).
Aspect 13. The method of any one of aspects 9 to 12, wherein the nucleophilic compound is a heteroatom-containing compound.
Aspect 14. The method of aspect 13, wherein the heteroatom-containing compound is a heterocycle or a cycle substituted with a heteroatom-containing moiety.
Aspect 15. The method of aspect 14, wherein the heterocycle is pyrrole, furan, thiophene, pyridine, indole, benzofuran, benzothiphene, imidazole, or derivatives thereof, each of which is optionally substituted.
Aspect 16. A pharmaceutical composition comprising a compound of any one of aspects 1 to 8, or a compound syntheized by the method of any one of aspects 8 to 15, and a pharmaceutically acceptable carrier, diluent, or vehicle.
Aspect 17. A method of treating a subject having or suspected of having an infectious disease, comprising: administering a therapeutically effective amount of a compound of any one of aspects 1 to 8, or a compound syntheized by the method of any one of aspects 8 to 15, or a pharmaceutical composition of aspect 16.
Aspect 18. A method of treating a subject having or suspected of having an infectious disease, comprising: administering a therapeutically effective amount of a compound of any one of aspects 1 to 8, or a compound syntheized by the method of any one of aspects 8 to 15, or a pharmaceutical composition of aspect 16, wherein said infectious disease is caused by a virus.
Aspect 19. The method of aspects 18, wherein said virus is a virus from the family Flaviviridae.
Aspect 20. The method of aspect 19, wherein the virus is from the genera Hepacivirus, Flavivirus, Pegivirus, or Pestivirus.
Aspect 21. The method of aspect 20, wherein said flavivirus is yellow fever virus (YFV), Japanese encephalitis virus (JEV), Tick-borne encephalitis virus (TBEV), Dengue virus (DENV), West Nile virus (WNV), Zika virus (ZIKAV), or any combination thereof.
Aspect 22. The method of aspect 18, wherein said virus is from the family Paramyxoviridae.
Aspect 23. The method of aspect 22, wherein said virus is from the genera Paramyxovirus, Pneumovirus, or Morbillivirus.
Aspect 24. The method of aspect 23, wherein Paramyxovirus is parainfluenza virus or mumpus virus.
Aspect 25. The method of aspect 23, wherein said Pneumovirus is respiratory syncytial virus (RSV).
Aspect 26. The method of aspect 23, wherein said Morbillivirus is measles virus.
Aspect 27. The method of any one of aspects 17 to 26, wherein said subject is a human, a domesticated animal, livestock, a laboratory animal, a non-human mammal, a non-human primate, a rodent, a bird, a reptile, an amphibian, or a fish.
Aspect 28. Use of a compound of any one of aspects 1 to 8, or a compound syntheized by the method of any one of aspects 8 to 15, or a pharmaceutical composition of aspect 16 for treating a subject having or suspected of having an infectious disease.
Aspect 29. Use of a compound of any one of aspects 1 to 8, or a compound syntheized by the method of any one of aspects 8 to 15, or a pharmaceutical composition of aspect 16 in the manufacture of a medicament for treating a subject having or suspected of having an infectious disease.
Aspect 30. Use of a compound of any one of aspects 1 to 8, or a compound syntheized by the method of any one of aspects 8 to 15, or a pharmaceutical composition of aspect 16 for treating a subject having or suspected of having an infectious disease, wherein said infectious disease is caused by a virus.
Aspect 31. Use of a compound of any one of aspects 1 to 8, or a compound syntheized by the method of any one of aspects 8 to 15, or a pharmaceutical composition of aspect 16 in the manufacture of a medicament for treating a subject having or suspect of having an infectious disease, wherein said infectious disease is caused by a virus.
Aspect 32. The use of aspect 30 or 31, wherein said virus is a virus from the family Flaviviridae.
Aspect 33. The use of aspect 32, wherein the virus is from the genera Hepacivirus, Flavivirus, Pegivirus, or Pestivirus.
Aspect 34. The use of aspect 33, wherein said flavivirus is yellow fever virus (YFV), Japanese encephalitis virus (JEV), Tick-borne encephalitis virus (TBEV), Dengue virus (DENV), West Nile virus (WNV), Zika virus (ZIKAV), or any combination thereof.
Aspect 35. The use of aspect 30 or 31, wherein said virus is a virus from the family Paramyxoviridae.
Aspect 36. The use of aspect 35, wherein said virus is from the genera Paramyxovirus, Pneumovirus, or Morbillivirus.
Aspect 37. The use of aspect 36, wherein Paramyxovirus is parainfluenza virus or mumpus virus.
Aspect 38. The use of aspect 36, wherein said Pneumovirus is respiratory syncytial virus (RSV).
Aspect 39. The use of aspect 36, wherein said Morbillivirus is measles virus.
Aspect 40. The use of any one of aspects 28 to 39, wherein said subject is a human, a domesticated animal, livestock, a laboratory animal, a non-human mammal, a non-human primate, a rodent, a bird, a reptile, an amphibian, or a fish.
Aspect 41. A method of inhibiting an RNA-dependent RNA polymerase (RdRP) of an RNA virus, the method comprising contacting the RdRP with a compound according to any one of aspects 1-8.
To gain a better understanding of the invention described herein, the following examples are set forth. It should be understood that these examples are for illustrative purposes only. Therefore, they should not limit the scope of this invention in anyway.
Aryl azides trapped ortho metallocarbene intermediates to generate indolenones possessing a reactive C-acylimine moiety, which reacted with added indole nucleophiles to afford a 2-(3-indolyl)indolin-3-one scaffold found in antiviral natural product isatisine A. This overall process occurred through a dual catalytic sequence at room temperature. Redox activation of a Cu(OTf)2 precatalyst by indole resulted in catalytically competent Cu(I) required for azide-metallocarbene coupling. A Bronsted acid that formed from Cu(OTf)2 reduction was responsible for catalysis of the C—C bond-forming indole addition step. This modular method allowed for rapid assembly of bis(indole) libraries, several of which demonstrated anti-infective activity against respiratory syncytial virus and Zika virus.
Reaction sequences proceeding through high-energy reactive intermediates, generated catalytically, can offer an approach for rapid construction of structurally complex products. Metallocarbenes generated from diazoketone precursors can be intercepted by a variety of nucleophilic heteroatom-containing functionalities to afford ylide intermediates that then undergo rearrangement processes. Use of an organoazide functionality to trap metallocarbene intermediates presents an alternative approach for strategic assembly of substituted heterocyclics. This process is catalyzed by inexpensive copper catalysts, with two equivalents of nitrogen gas being the only by-products generated, and C-acylimines formed from this coupling are subject to further reaction via in situ intermolecular nucleophilic trapping to give highly substituted indolin-3-one products through a one-pot cascade process. This process delivers a nucleophilic addend adjacent to one or more carbonyl moieties in umpollung fashion. Indoles were found to be an especially effective class of traps, and it was noted that there are structural similarities between these bis(indole) adducts to isatisine A, a naturally occurring compound with antiviral properties.
Given the modular nature of the diazoketone→metallocarbene→C-acylimine→indole adduct cascade sequence, construction of a library of unnatural Isatisine A-inspired bis(indole) compounds lacking a ribose-derived ‘eastern’ fragment was undertaken (see
Experimental Details
Reactions were carried out in oven (130° C.) or flame-dried glassware under a positive argon atmosphere unless otherwise stated. Transfer of anhydrous reagents was accomplished with oven-dried syringes or cannulae. Solvents were distilled before use: acetonitrile (CH3CN), dichloromethane (DCM) and dichloroethane (DCE) from calcium hydride, toluene (PhMe) from sodium metal, diethyl ether (Et2O) and tetrahydrofuran (THF) from sodium metal/benzophenone ketyl. Thin layer chromatography was performed on glass plates pre-coated with 0.25 mm silica gel with fluorescent indicator UV254 (Rose Scientific). Flash chromatography columns were packed with 230-400 mesh silica gel (Silacycle). Proton nuclear magnetic resonance spectra (1H NMR) were recorded at 400 MHz, or 500 MHz and coupling constants (J) are reported in Hertz (Hz). Carbon nuclear magnetic resonance spectra (13C NMR) were recorded at 125 MHz, as proton decoupled or as attached proton test (APT). Chemical shifts are reported on a 6 scale (ppm) and referenced to residual solvent peaks: CDCl3 (7.26 ppm, 1H; 77.06 ppm, 13C), d6-DMSO (2.49 ppm, 1H; 39.5 ppm, 13C), and d2-DCM (5.32 ppm, 1H; 53.5 ppm, 13C).
All reagents were purchased from commercial suppliers: Sigma Aldrich, AK Scientific, and Acros, at purity greater than (95%). Nuclear Magnetic Resonance (NMR) measurements (chemical shift, integration, coupling constants, relative peak intensity) were performed using Agilent 400 MHz, Varian 400 MHz, or Varian 500 MHz NMR instruments. Mass measurements were acquired using Agilent Technologies 1100 MSD (Single Quadruple), Waters (Micromass) Q-TOF Premier (Quadruple TOF), Kratos Analytical MS-50G, Agilent Technologies 6220 oaTOF, or AB Sciex QTRAP 2000 instruments. UV-VIS spectra were acquired using Cary UV-Vis, or Hewlett Packard 8453 UV-VIS Spectrometers. Elemental Analysis was performed using Thermo Flash 2000 Elemental Analyzer. Infrared spectra were acquired using Mattson Galaxy Series FT-IR 3000, Nicolet Magna 750 FTIR Spectrometer equipped a Nic-Plan FTIR Microscope, or Thermo Nicolet 8700 FTIR Spectrometer equipped with a Continuum FTIR Microscope. Differential Scanning calorimetry and Thermogravimetric analysis were performed using Perkin Elmer Pyris 1, or Mettler Toledo TGA/DSC instruments.
For virology procedures: MEM media (HyClone, GE Healthcare Life Sciences); Opti-MEM media (Gibco, Thermofisher); fetal bovine serum (FBS) (Gibco, Thermofisher); DMEM (HyClone, GE Healthcare Life Sciences); Methanol (HPLC Grade, Fisher Chemical); Acetone (ACS Certified, Fisher Chemical); Goat anti-RSV polyclonal antibody (Meridian Life Science B65860G); Alexafluor 647 (LifeTech A-21469); MTT ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)); Phosphate buffered saline (PBS) (Gibco, Thermofisher); Dimethyl sulfoxide (DMSO) (ACS Certified, Fisher Chemical); [a 32-P] UTP was purchased from PerkinElmer. All remaining reagents (actinomycin D, lysolecithin, Tris-acetate, Magnesium acetate, Potassium acetate, DTT, spermidine, creatine phosphatase, aprotinin, creatine phosphokinase, ATP, GTP, CTP, UTP, and RNaseH) were purchased from Sigma Aldrich at purity greater than (95%).
Substrate Preparation
Preparation of compounds 1a, 1b, 1d, and 1g have been reported previously by Bott, T. M.; Atienza, B. J.; West, F. G. RSC Adv. 2014, 4, 31955-31959.
Dichloromethane (DCM, 35 mL) was added to a conical flask containing 4,5-dimethoxy-2-azido benzoic acid (1.0 g, 4.5 mmol) and the suspension was cooled to 0° C. before addition of methyl acetate (1.0 equiv) and trichloroacetyl chloride (1.2 equiv). This solution was slowly transferred via cannula to a suspension of NaH (1.2 equiv) in DCM (10-20 mL) at 0° C. After stirring at 0° C. for 15 min, the solution was cooled to −45° C. before addition of 1-methylimidazole (1.2 equiv). The solution was then stirred for an additional 10 min at −45° C. before slowly adding TiCl4 (3.4 equiv) followed by NEt3 (4.0 equiv). The dark red-brown solution was kept at −78° C. for 30 min before being warmed to 0° C. and kept for 1 h and subsequently quenched with water (30 mL). The organic layer was separated and aqueous layer was washed 3x with equal portions of DCM. The combined organic layers were washed with an equivalent volume of water and brine, dried over MgSO4, filtered, and concentrated under reduced pressure. Crude product was partially purified by flash chromatography to afford an orange oil whose Rf was about 0.3 (7:3 hexanes:EtOAc). The orange oil was concentrated and added to a stirred solution of triethylamine (1.2 equiv) in CH3CN (20 mL). Tosyl azide (1.0 equiv) in CH3CN (10 mL) was transferred via cannula into the flask and the reaction was left to stir overnight. Concentration under reduced pressure followed by purification via flash chromatography (silica gel, 8:2 hexanes:Et0Ac→7:3 hexanes:EtOAc slowly added in gradient) furnished lc as a yellow oil in 50% yield (from starting 4,5-dimethoxy-2-azidobenzoic acid).
Rf=0.25 (7:3 hexanes:EtOAc); IR (cast film) 2978, 2134, 1711, 1695, 1565, 1292 cm−1; 1H NMR (500 MHz, CDCl3) δ 6.87 (s, 1H), 6.62 (s, 1H), 3.94 (s, 3H), 3.86 (s, 3H), 3.78 (s, 3H); 13C NMR (125 MHz, CDCl3) δ 184.5, 161.2, 152.4, 146.4, 131.4, 121.9, 111.6, 101.5, 56.3, 56.2, 52.3; HRMS calc'd for C12H11N5O5Na [M+Na]+328.0652, found 328.0651. (NB.: 13C signal for the diazo carbon was not detected due to quadrupolar broadening.).
Prepared analogously to 1c using 3-nitro-2-azido benzoic acid in place of 2-azido-4,5-dimethoxybenzoic acid. Isolated as a brown oil in 23% yield: Rf=0.19 (7:3 hexanes:EtOAc); IR (cast film) 3022, 2141, 1732, 1694, 1637, 1567, 1293 cm−1; 1H NMR (500 MHz, CDCl3) δ 8.14 (dd, J=8.2, 1.6 Hz, 1H), 7.51 (dd, J=7.6, 1.6 Hz, 1H), 7.38 (dd, J=8.2, 7.6 Hz, 1H), 3.76 (s, 3H); 13C NMR (125 MHz, CDCl3) δ 184.2, 160.5, 135.8, 132.7, 132.5, 132.4, 127.8, 125.8, 52.6; LC-MS calc'd for C1oH7N6O5 [M+H]+291.1, found 291.1. (NB.: (a) preparation for this starting material was limited to 70-100 mg; (b) 13C signal for the diazo carbon was not detected due to quadrupolar broadennin).
Prepared analogously to lc using 3-azido-2-napthoic acid in place of 2-azido-4,5-dimethoxybenzoic acid.
Isolated as a brown oil in 51% yield: Rf=0.68 (7:3 hexanes:EtOAc); IR (cast film) 2971, 2136, 1724, 1693, 1633, 1567, 1290 cm−1; 1H NMR (500 MHz, CDCl3) δ 7.83-7.76 (m, 3H), 7.53 (br s, 1H), 7.54 (ddd, J=8.2, 6.9, 1.2 Hz, 1H), 7.24 (ddd, J=8.2, 7.0, 1.2 Hz, 1H), 3.74 (s, 3H); 13C NMR (125 MHz, CDCl3) δ 185.6, 160.9, 135.2, 134.7, 130.3, 129.5, 128.9, 128.7, 128.4, 126.7, 126.2, 115.6, 52.3; HRMS calc'd for C14H9N5O3Na [M+Na]+318.0597, found 318.0594. (NB.: 3C signal for the diazo carbon was not detected due to quadrupolar broadening.).
Prepared analogously to lc using vinyl acetate in place of methyl acetate, and using 2-azido benzoic acid in place of 2-azido-4,5-dimethoxy benzoic acid.
Isolated as a yellow oil in 46% yield: Rf=0.51 (7:3 hexanes:EtOAc); IR (cast film) 2924, 2931, 2136, 1724, 1684 cm−1; 1H NMR (400 MHz, CDCl3) 6 7.47 (ddd, J=8.1, 7.5, 1.6 Hz, 1H), 7.28-7.25 (m, 1H), 7.17-7.12 (m, 3H), 4.70 (dd, J=13.9, 2.0 Hz, 1H), 4.54 (dd, J=6.2, 2.0 Hz, 1H); 13C NMR (125 MHz, CDCl3) δ 185.3, 157.7, 140.1, 137.9, 132.2, 130.2, 128.6, 124.8, 118.3, 98.5; HRMS calc'd for C11H7N5O3Na [M+Na]+280.0441, found 280.0439. (NB.: 13C signal for the diazo carbon was not detected due to quadrupolar broadening.).
Prepared analogously to lc using (+)-menthyl acetate in place of methyl acetate, and using 2-azido-benzoic acid in place of 2-azido-4,5-dimethoxy benzoic acid.
Isolated as a yellow oil in 48% yield: Rf=0.64 (7:3 hexanes:EtOAc); [α]D20:−67.76 (c=0.52, DCM) IR (cast film) 2956, 2928, 2870, 2130, 1719, 1691, 1302, 958 cm−1; 1HNMR (500 MHz, CDCl3) δ 7.45 (ddd, J=8.1, 7.4, 1.7 Hz, 1H), 7.30-7.28 (m, 1H), 7.19-7.14 (m, 2H), 4.70 (ddd, J=10.9, 10.9, 4.4, 1H), 2.00-1.96 (m, 1H), 1.71-1.58 (m, 3H), 1.44-1.36 (m, 1H), 1.15-0.93 (m, 2H), 0.87 (d, J=6.5 Hz, 3H), 0.83 (d, J=7.0 Hz, 3H), 0.87-0.83 (m, 2H), 0.72 (d, J=7.0 Hz, 3H); 13C NMR (125 MHz, CDCl3) δ 185.9, 160.3, 137.6, 131.7, 130.8, 128.3, 124.7, 118.1, 75.9, 46.9, 40.9, 34.0, 31.3, 26.4, 23.4, 21.9, 20.7, 16.3; HRMS calc'd for C19H23N5O3Na [M+Na]+392.1693, found 392.1692. (NB.: 13C signal for the diazo carbon was not detected due to quadrupolar broadening.).
Prepared analogously to lc using 2-azido-3-bromo benzoic acid in place of 2-azido-4,5-dimethoxybenzoic acid. Product was obtained in 43% yield (unoptimized due to procedure being run only once, on a sufficient scale to permit its use in multiple coupling experiments).
Isolated as a yellow oil: Rf=0.64 (3:7, EtOAc:hexanes); IR (cast film) 3026, 2955, 2111, 1729, 1634, 1585, 1565 cm−1; 1H NMR (500 MHz, CDCl3): δ 7.35-7.33 (m, 2H), 7.18 (d, J=8.5 Hz, 1H), 3.78 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 184.7, 160.7, 139.2, 129.7, 129.2, 128.0, 125.7, 121.5, 52.4; HRMS calc'd for C10H6BrN5NaO3 [M+Na]+345.9546, found 345.9549. (NB.: 13C signal for the diazo carbon was not detected due to quadrupolar broadening.).
A solution of LiHMDS (6.2 mL of 1M THF solution; 6.2 mmol) was added to a solution of 2-azido-acetophenone (1.0 g, 6.2 mmol) in 10 mL THF at −78° C. The mixture was allowed to stir for 30 min. Then, a solution of benzoyl cyanide (822 mg, 6.20 mmol, dissolved in 10 mL THF) was added dropwise. The mixture was allowed to stir for 1 h then quenched with saturated NH4Cl (15 mL). The mixture was diluted with diethyl ether (10 mL). The organic layer was separated and washed with water (20 mL, 3×). The organic layer was washed with brine and dried with MgSO4. The solution was concentrated under pressure to afford yellow oil. The yellow oil was purified using flash column chromatography eluting 10% % EtOAc in hexanes to furnish 1.13 g (69%) of intermediate β-diketone (isolated as an enol) as a yellow oil:
Rf=0.63 (7:3 hexanes:EtOAc); IR (cast film) 3064, 2420, 2125, 1599, 1281, 1604, 778, 1126, 750 cm−1; 1H NMR (500 MHz, CDCl3) δ 8.01-7.87 (m, 3H), 7.59-7.48 (m, 4H), 7.28-6.99 (m, 2H), 7.00 (s, 1H); 13C NMR (125 MHz, CDCl3,) δ 185.5, 184.4, 138.3, 135.4, 132.6, 132.5, 130.4, 128.7, 128.3, 127.3, 125.0, 119.3, 98.3; HRMS calc'd for C11H11N3O2 [M]+265.0851, found: 294.0849. (NB.: enol H was not detected.)
Triethylamine (715 μL, 5.1 mmol) was added to a stirred solution of the intermediate diketone (500 mg, 1.89 mmol) in CH3CN (8 mL). Tosyl azide (400 mg, 2.00 mmol) in CH3CN (5 mL) was transferred via cannula into the flask and the reaction was left to stir overnight. Concentration under reduced pressure followed by purification via flash chromatography (silica gel, 7:3 hexanes:EtOAc) resulted in a quantitative yield of 11(551 mg) as a pale yellow oil. To prevent decomposition, product was stored under Ar in a freezer in a foil wrapped flask. Under these conditions the compound was stable over several months.
Rf=0.71 (7:3 hexanes:EtOAc); IR (cast film) 3053, 2451, 2132, 1641, 1283, 1607, 779, 1136, 754 cm−1; 1H NMR (500 MHz, CDCl3) δ 7.55-7.53 (m, 2H), 7.43-7.28 (m, 5H), 7.13 (ddd, J=7.6, 7.6, 0.8 Hz, 1H), 6.92 (d, J=8.0 Hz, 1H); 13C NMR (125 MHz, CDCl3) δ 185.9, 184.9, 137.5, 136.7, 132.5, 132.5, 130.0, 129.5, 128.2, 128.0, 125.0, 118.2, 98.3; LC-MS calc'd for C15H9N5O2 [M+H]+291.2, found: 291.2.
Dual-Catalytic Formation of Bis(Indole) Products 2 and 5
Representative Procedure for Small-Scale Synthesis of 2a (Method A):
A solution of diazoazide 1a (0.250 g, 1.02 mmol) in DCM (25 mL) was added to a solution of indole (0.239 g, 2.04 mmol) and Cu(OTf)2 (37 mg, 0.102 mmol) in DCM (25 mL) at room temperature via syringe pump over 1 h. The reaction mixture turned light green over 1-2 h and slowly turned dark brown over 24 h. Once addition was complete, the reaction was monitored by TLC for consumption of 1a, (for some diazoazide starting materials, the reaction mixture was heated further at reflux for 10-15 minutes to ensure completion). Upon consumption of 1a, the reaction mixture was concentrated under reduced pressure, purified by flash chromatography (silica gel, 7:3 hexanes:EtOAc), and recrystallization (MeOH) (See Example 2 for more details). Total isolated yield of 2a was 0.287 g (92%), as a combined yellow amorphous powder and crystals. Suitable single crystals of 2a for X-ray diffraction were grown from 1:1 MeOH-EtOAc via slow evaporation of solvent.
Representative Procedure for Gram-Scale Synthesis of 2a (Method B):
A solution of diazoazide 1a (5.00 g, 20.4 mmol) in DCM (150-250 mL) was added to a solution of indole (4.78 g, 40.8 mmol) and Cu(OTf)2 (740 mg, 2.04 mmol) in DCM (150-250 mL) at room temperature via syringe pump. The reaction mixture turned light green over 2 h and slowly turned dark brown with suspended green solid over 24 h. The reaction was monitored by TLC for consumption of 1a. Upon consumption of 1a, the suspended solid was filtered from the reaction mixture to afford green needles of 2a. The green needles were directly recrystallized with minimum amount of ethyl acetate (hot), followed by slow addition of pentane or hexane to furnish adduct 2a as a fine dark yellow solid in 74% (69-78%) yield. The mother liquor contained ca. 10% of the product and could still be purified by flash chromatography (silica gel, 7:3 Hexanes:EtOAc) to afford yellow oil, which precipitated when left standing at −4° C. overnight.
Following Method A, 0.231 g (74%) of dark yellow crystalline 2a was isolated after recrystallization. Concentration of mother liquor afforded another 0.056 g (18%) of yellow/brown amorphous powder. Combined total of 2a was 0.287 g (92%), m.p. ˜110° C. (typical for the yellow/brown powder), m.p.=225-226° C. (crystals); IR (cast film) 3392, 3059, 2953, 1726, 1697, 1491, 1434, 1214, 748 cm1; 1H NMR (500 MHz, CDCl3) δ 8.22 (br s, 1H), 7.70 (d, J=7.7 Hz, 1H), 7.59 (d, J=8.3 Hz, 1H), 7.53 (app td, J=8.3, 1.3 Hz, 1H), 7.40 (d, J=2.4 Hz, 1H), 7.37 (d, J=8.1 Hz, 1H), 7.20 (app t, J=7.2 Hz, 1H), 7.11 (app t, J=7.9 Hz, 1H), 7.01 (d, J=8.3 Hz, 1H), 6.93 (app t, J=7.7 Hz, 1H), 5.73 (br s, 1H), 3.80 (s, 3H); 13C NMR (125 MHz, CDCl3) 193.3, 168.5, 159.5, 140.4, 136.5, 127.8, 125.3, 123.5, 122.8, 121.4, 120.5, 119.4, 115.1, 112.6, 111.7, 111.2, 73.0, 53.9; HRMS calc'd for C18H15N2O3 [M+H]+307.1077, found 307.1078; Elemental analysis calc'd for C18H14N2O3: 70.58% C, 4.61% H, 9.15% N, 15.67% O, found, 70.48% C, 4.62% H, 9.13% N. 1H-NMR spectral data were in good agreement with those reported in the literature [Jessing, M.; Barran, P. S. Heterocycles 2011, 82, 1739-1745]. 13C-NMR spectral varied by up to 0.6 ppm for some 13C NMR resonances. There were also additional three carbon resonances reported here not included in the earlier data. Similar to 1H-NMR spectral characterizations, IR wavenumbers were likewise in good agreement.
(Method A) Isolated as yellow cubic crystals in 89% yield (using methanol as crystallizing solvent): m.p.=215-216° C. (reported for powder 82° C. [Jessing, M.; Barran, P.S. Heterocycles 2011, 82, 1739-1745] and [Bolt, T. M.; Atienza, B. J.; West, F. G. RSC Adv. 2014, 4, 31955-31959]); Rf=0.31 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3366, 3051, 1744, 1702, 1616, 1485, 1293, 742 cm−1; 1H NMR (500 MHz, CDCl3): δ 7.73 (d, J=7.0 Hz, 1H), 7.62 (d, J=7.5 Hz, 1H), 7.54 (app t, J=6.5 Hz, 1H), 7.34 (d, J=8.0 Hz, 1H), 7.30 (d, J=5.5 Hz, 1H), 7.27 (app t, J=7.0 Hz, 1H), 7.14 (app t, J=7.0 Hz, 1H), 7.00 (d, J=8.0 Hz, 1H), 6.96 (app t, J=6.8 Hz, 1H), 5.34 (br s, 1H), 3.84 (s, 3H), 3.77 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 195.2, 169.1, 161.0, 137.9, 137.4, 128.0 126.0, 125.4, 122.3, 120.4, 120.0, 119.9, 119.5, 113.6, 109.8, 109.8, 72.4, 53.8, 32.9; HRMS calc'd for C19H17N2O3 [M+H]+321.1234, found 321.1233.
(Method A) Isolated as a yellow crystalline solid in 93% combined yield (Method B: 77-83% yield if direct recrystallization from crude reaction mixture using methanol as the solvent): m.p.=181-183° C.; Rf=0.40 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3370, 3051, 2920, 1742, 1704, 1616, 1486, 743 cm−1; 1H NMR (500 MHz, CDCl3): δ 7.72 (d, J=7.7 Hz, 1H), 7.64 (d, J=7.9 Hz, 1H), 7.55 (ddd, J=8.3, 7.2, 1.3 Hz, 1H), 7.42 (s, 1H), 7.32-7.25 (m, 4H), 7.20 (app td, J=8.3, 1.0 Hz, 1H), 7.15-7.11 (m, 3H), 7.02 (d, J=8.3 Hz, 1H), 6.95 (app t, J=7.9 Hz, 1H), 5.79 (br s, 1H), 5.30 (s, 2H), 3.83 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 194.7, 169.0, 161.0, 137.9, 137.0, 136.9, 128.8, 127.7, 127.6, 126.9, 126.3, 125.4, 122.4, 120.4, 120.2, 119.9, 119.8, 113.6, 110.5, 110.3, 72.4, 53.8, 50.4; HRMS calc'd for C25H20N2O3Na [M+Na]+419.1366, found: 419.1363; Fluorescent properties in methanol solution: excitation wavelengths at 290 nm and 395 nm, emission wavelength at 500 nm (both excitation wavelength).
(Method A) Isolated as a bright yellow microcrystals in 88% yield; m.p.=127° C. (decomp.); Rf=0.28 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3360, 3062, 2950, 1710, 1617, 1487, 1239, 748 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.29 (br s, 1H), 7.66 (br d, J=8.1 Hz, 1H), 7.57 (ddd, J=8.4, 7.2, 1.5 Hz, 1H), 7.48-7.46 (m, 2H), 7.42-7.37 (m, 3H), 7.35 (d, J=8.1 Hz, 1H), 7.23 (d, J=8.3 Hz, 1H), 7.20 (ddd, J=8.3, 7.2, 1.1 Hz, 1H), 7.05 (ddd, J=8.3, 7.2, 1.1 Hz, 1H), 7.00 (d, J=8.3 Hz, 1H), 6.97 (app t, J=7.9 Hz, 1H), 5.62 (br s, 1H), 3.24 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 195.1, 168.8, 160.8, 138.0, 137.9, 135.5, 132.5, 129.6, 128.8, 128.4, 126.8, 125.2, 122.7, 120.6, 120.4, 120.2, 119.7, 113.4, 111.2, 108.2, 73.3, 53.2; HRMS calc'd for C24H19N2O3[M+H]−383.1390, found 383.1385.
(Method A) Isolated as a light green crystalline solid in 71% yield; m.p.=242-243° C.; Rf=0.23 (yellow spot, 7:3 hexanes:EtOAc); IR (microscope) 3365, 3006, 2953, 1726, 1706, 1610, 1488, 1219, 792 cm−1; 1H NMR (500 MHz, d6-DMSO): 6 7.60 (br s, 1H), 7.55 (d, J=7.9 Hz, 1H), 7.51 (app t, J=8.3 Hz, 1H), 7.45 (d, J=7.9 Hz, 1H), 7.27 (d, J=7.5 Hz, 1H), 7.10 (d, J=2.8 Hz, 1H), 7.06 (app t, J=7.9 Hz, 1H), 7.04 (d, J=8.3 Hz, 1H), 6.81 (app t, J=7.5 Hz, 1H), 3.61 (s, 3H) (Note: one NH proton was not detected); 13C NMR (125 MHz, d6-DMSO): δ 195.9, 169.9, 162.2, 138.6 (2×), 126.4, 125.1, 125.0, 124.0, 123.3, 118.9, 118.0, 114.0, 113.0, 112.1, 111.4, 72.4, 53.5; HRMS calc'd for C18H1479BrN2O3[M+H]+385.0182, found 385.0182; Elemental analysis calc'd for C18H13BrN2O3: 56.12% C, 3.40% H, 7.27% N; found, 55.98% C, 3.41% H, 7.24% N.
(Method A) Isolated as bright yellow crystals in 78% yield; m.p.=186-188° C.; Rf=0.11 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3301, 3060, 2947, 1742, 1685, 1487, 1232, 1218, 756 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.19 (br s, 1H), 7.74 (dd, J=7.9, 0.6 Hz, 1H), 7.56 (app td, J=8.3, 1.3 Hz, 1H), 7.38 (d, J=2.6 Hz, 1H), 7.30-7.26 (m, 2H), 7.06-7.04 (m, 2H), 6.98 (app t, J=7.2 Hz, 1H), 6.89 (dd, J=8.8, 2.4 Hz, 1H), 5.71 (br s, 1H), 3.81 (s, 3H), 3.77 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 194.9, 169.0, 161.0, 154.4, 137.9, 131.7, 125.8, 125.4, 124.3, 120.4, 120.0, 113.5, 112.7, 112.3, 111.2, 101.5, 72.4, 55.8, 53.7; HRMS calc'd for C19H17N2O4 [M+H]+337.1183, found 337.1185.
(Method A, with following modification: after addition of diazo-azide 1a by syringe pump, reaction mixture was heated to reflux and stirred at that temperature until consumption of 1a was observed) Isolated as a yellow oil in 46% yield; Rf=0.40 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3363, 3065, 2954, 1748, 1711, 1617, 1488, 1242, 751 cm−1; 1H NMR (500 MHz, CDCl3): δ 7.69 (d, J=7.9 Hz, 1H), 7.55 (dd, J=7.7, 0.6 Hz, 1H), 7.54 (ddd, J=8.3, 7.2, 1.3 Hz, 1H), 7.45 (dd, J=8.1, 0.6 Hz, 1H), 7.28 (app td, J=7.2, 1.3 Hz, 1H), 7.21 (app td, J=7.9, 1.1 Hz, 1H), 7.03 (d, J=8.3 Hz, 1H), 6.95 (overlapped app td, J=7.7, 0.6 Hz, 1H), 6.94 (overlapped s, 1H), 5.74 (br s, 1H), 3.85 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 191.3, 166.6, 161.2, 155.0, 150.9, 138.2, 127.7, 125.7, 124.9, 123.1, 121.6, 120.8, 119.3, 113.7, 111.3, 105.6, 71.9, 54.2; HRMS calc'd for C18H13NO4Na [M+Na]+330.0737, found 330.0739.
(Method A) Isolated as a yellow oil in 68% yield: Rf=0.34 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3426, 3390, 3056, 2954, 1726, 1697, 1488, 1233, 751 cm−1; 1H NMR (500 MHz, CDCl3): δ 9.43 (br s, 1H), 7.63 (d, J=7.9 Hz, 1H), 7.56 (apparent (app) td, J=7.5, 1.5 Hz, 1H), 7.09 (d, J=7.5 Hz, 1H), 6.94 (app td, J=7.2, 0.7 Hz, 1H), 6.87 (app td, J=2.6, 1.7 Hz, 1H), 6.33 (ddd, J=3.5, 2.7, 1.7 Hz, 1H), 6.19-6.14 (m, 1H), 5.68 (br s, 1H), 3.84 (s, 3H); 13C NMR (125 MHz, CDCl3): 8 193.2, 167.6, 161.6, 138.0, 125.8, 124.8, 120.7, 119.2, 118.8, 113.6, 108.2, 106.3, 71.8, 53.9; HRMS calc'd for C14H13N2O3[M+H]+257.0921, found 257.0915.
(Method A, with following modification: after addition of diazoazide 1a by syringe pump, the reaction mixture was heated to reflux and stirred at that temperature until consumption of 1a was observed) Isolated as a yellow oil in 48% yield; Rf=0.37 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3362, 3125, 2954, 1748, 1710, 1617, 1488, 1233, 751 cm−1; 1H NMR (500 MHz, CDCl3): δ 7.69 (d, J=7.2 Hz, 1H), 7.55 (app td, J=7.2, 0.9 Hz, 1H), 7.44 (dd, J=1.8, 0.9 Hz, 1H), 7.03 (d, J=8.3 Hz, 1H), 6.96 (app t, J=7.7 Hz, 1H), 6.56 (d, J=3.3 Hz, 1H), 6.42 (dd, J=3.3, 1.9 Hz, 1H), 5.66 (br s, 1H), 3.86 (s, 3H); 13C NMR (125 MHz, CDCl3): 6 191.8, 166.9, 161.1, 148.4, 143.3, 138.1, 125.7, 120.7, 119.3, 113.6, 110.7, 108.8, 71.7, 54.1; HRMS calc'd for C14H12NO4[M+H]+258.0761, found 258.0764.
(Method A, with following modification: after addition of diazoazide 1a by syringe pump, reaction mixture was heated to reflux and stirred at that temperature until consumption of 1a was observed) Isolated as a yellow oil in 41% yield; Rf=0.42 (7:3 hexanes:EtOAc); IR (cast film) 3360, 3071, 2952, 1747, 1709, 1616, 1486, 1231, 754 cm−1; 1H NMR (500 MHz, CDCl3): δ 7.62 (ddd, J=7.2, 1.4, 0.7 Hz, 1H), 7.54 (ddd, J=8.4, 7.2, 1.4 Hz, 1H), 7.49 (dd, J=3.7, 1.3 Hz, 1H), 7.24 (dd, J=5.1, 1.3 Hz, 1H), 7.07 (d, J=8.3 Hz, 1H), 7.03 (dd, J=5.1, 3.7 Hz, 1H), 6.95 (app td, J=7.9, 0.7 Hz, 1H), 5.79 (br s, 1H), 3.85 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 192.2, 167.4, 160.9, 138.8, 137.9, 127.6, 125.9, 125.7, 125.5, 121.1, 119.0, 113.7, 73.4, 54.1; HRMS calc'd for C14H12NSO3[M+H]+274.0532, found 274.0536.
(Method A) Isolated as yellow crystals in 86% yield; m.p.=192-193° C.; Rf=0.16 (yellow spot, 7:3 hexanes:EtOAc); IR(cast film) 3371, 3057, 2952, 1739, 1702, 1608, 1459, 1238, 745 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.40 (br s, 1H), 7.58-7.55 (m, 2H), 7.37-7.31 (m, 3H), 7.18 (ddd, J=7.9, 7.0, 1.2 Hz, 1H), 7.10 (ddd, J=8.1, 7.1, 1.0 Hz, 1H), 6.89 (app t, J=7.5 Hz, 1H), 5.53 (br s, 1H), 3.79 (s, 3H), 2.30 (s, 3H); 13C NMR (125 MHz, CDCl3) δ 195.2, 169.2, 160.3, 138.0, 136.6, 125.5, 123.7, 122.8 (2×), 122.7, 120.6, 120.4, 119.5 (2×), 111.8, 111.7, 72.6, 53.8, 15.8; HRMS calc'd for C19H17N2O3 [M+H]+321.1234, found: 321.1233.
(Method A) Isolated as yellow crystals in 84% yield (using MeOH as the recrystallization solvent); m.p.=170-171° C.; Rf=0.35 (yellow spot, 7:3 hexanes:EtOAc); IR(cast film) 3359, 3055, 2950, 1746, 1705, 1608, 1460, 1241, 743 cm−1; 1H NMR (500 MHz, CDCl3) δ 7.61 (2 overlapping app t, J=8.2 Hz, 2H), 7.39-7.26 (m, 4H), 7.15 (app t, J=7.9 Hz, 1H), 6.91 (app t, J=7.3 Hz, 1H), 5.58 (br s, 1H), 3.84 (s, 3H), 3.79 (s, 3H), 2.33 (s, 3H); 13C NMR (125 MHz, CDCl3) δ 195.2, 169.2, 160.2, 137.9, 137.4, 128.1, 126.1, 122.8, 122.7, 122.2, 120.5, 119.9, 119.5, 119.5, 110.0, 109.8, 72.5, 53.8, 32.9, 15.8; HRMS calc'd for C20H19N2O3 [M+H]+335.1390, found: 335.1388.
(Method A) Isolated as a yellow powder in 94% yield (using methanol as recrystallization solvent): m.p.=184-186° C.; Rf=0.16 (7:3 hexanes:EtOAc); IR (cast film) 3349, 3015, 2942, 1748, 1712, 1623, 1451, 1239, 731 cm−1; 1H NMR (500 MHz, CDCl3) δ 7.61 (d, J=7.8 Hz, 1H), 7.60 (s, 1H), 7.28-7.09 (m, 10H), 6.48 (br s, 1H), 5.28 (s, 2H), 3.92 (s, 3H), 3.87 (s, 3H), 3.82 (s, 3H); 13C NMR (125 MHz, CDCl3) δ 192.7, 169.4, 158.9 (2×), 145.2, 136.9 (2×), 128.7, 127.6 (2×), 126.8, 126.4, 122.3, 120.1, 119.5, 111.5, 110.8, 110.3, 104.5, 95.8, 73.0, 56.3, 56.2, 53.7, 50.3; HRMS calc'd for C27H25N2O3 [M+H]+457.1685, found: 457.1683.
(Method A) Isolated as a yellow powder in 89% yield: m.p.=236-238° C.; Rf=0.19 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3364, 3052, 2952, 1746, 1711, 1610, 1242, 741 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.21 (br s, 1H), 7.61 (d, J=8.3 Hz, 1H), 7.55 (d, J=8.1 Hz, 1H), 7.40-7.37 (m, 2H), 7.22 (app t, J=7.7 Hz, 1H), 7.12 (app t, J=8.1 Hz, 1H), 7.00 (d, J=1.1 Hz, 1H), 6.90 (dd, J=8.3, 1.7 Hz, 1H), 5.77 (br s, 1H), 3.81 (s, 3H); 13C NMR (125 MHz, CDCl3) δ 193.2, 168.6, 161.2, 144.5, 136.5, 126.5, 125.3, 123.5, 122.9 121.2, 120.6, 119.4, 118.3, 113.4, 111.7, 111.3, 72.8, 53.9; HRMS calc'd for C18H13ClN2O3Na [M+Na]+363.0507, found: 363.0508; Elemental analysis calc'd for C18H13ClN2O3: 63.45% C, 3.85% H, 8.22% N; found, 63.35% C, 3.87% H, 8.13% N.
(Method A) Isolated as bright yellow crystals in 76% yield (using methanol as recrystallization solvent): m.p.=194-195° C.; Rf=0.35 (7:3 hexanes:EtOAc); IR (cast film) 3364, 3052, 2952, 1746, 1711, 1610, 1242, 741 cm−1; 1H NMR (500 MHz, CDCl3): δ 7.61 (d, J=8.0 Hz, 1H), 7.52 (ddd, J=8.0, 1.8, 1.0 Hz, 1H), 7.31 (ddd, J=8.3, 1.8, 1.1 Hz, 1H), 7.26-7.22 (m, 2H), 7.10 (ddd, J=8.1, 7.0, 1.1 Hz, 1H), 6.99 (dd, J=1.7, 0.5 Hz, 1H), 6.89 (dd, J=8.3, 1.7 Hz, 1H) 5.80 (br s, 1H), 3.81 (s, 3H), 3.75 (s, 3H); 13C NMR (125 MHz, CDCl3) δ 193.3, 168.6, 161.1, 144.5, 137.3, 128.0, 126.4, 125.8, 122.3, 121.1, 120.1, 119.3, 118.1, 113.3, 109.8, 109.3, 72.7, 53.9, 32.9; HRMS calc'd for C19H1635ClN2O3 [M+H]+355.0844, found: 355.0846.
(Method A) Isolated as bright yellow crystals in 90% yield (80% yield from direct recrystallization of crude and using methanol as recrystallization solvent): m.p.=174-175° C.; Rf=0.42 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3364, 3052, 2952, 1746, 1711, 1610, 1242, 741 cm−1; 1H NMR (500 MHz, CDCl3): δ 7.62 (app t, J=8.3 Hz, 2H), 7.40 (s, 1H), 7.31-7.26 (m, 4H), 7.20 (app td, J=7.0, 0.9 Hz, 1H), 7.14-7.11 (m, 3H), 6.97 (d, J=1.7 Hz, 1H), 6.89 (dd, J=8.3, 1.7 Hz, 1H) 5.96 (br s, 1H), 5.26 (s, 2H), 3.81 (s, 3H); 13C NMR (125 MHz, CDCl3) δ 193.3, 168.6, 161.1, 144.4, 136.9, 136.8 128.8, 127.7, 127.6, 126.8, 126.4, 126.1, 122.5, 120.9, 120.2, 119.7, 118.0, 113.2, 110.4, 110.0, 72.8, 53.8, 50.3; HRMS calc'd for C25H1935ClN2NaO3 [M+Na]+453.0976, found: 453.0984.
(Method A; Note: due to concerns about potential hazards associated with starting material, this reaction was carried out on 20 mg scale. As a result, only partial characterization was obtained.) Light yellow solid in ca. 14% yield (corrected for impurity from EtOAc and water present): Rf=0.24 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3324, 3048, 2956, 1738, 1714, 1610, 1251, 731 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.38 (dd, J=8.2, 1.2 Hz, 1H), 8.01 (s, 1H), 8.00-7.98 (m, 2H), 7.56 (dd, J=8.1, 1.0 Hz, 1H), 7.38 (br s, 1H), 7.29-7.12 (m, 5H), 6.95 (dd, J=8.2, 7.3 Hz, 1H), 5.26 (s, 2H), 3.83 (s, 3H); LC-MS calc'd for C25H19N3O5 [M+H]+442.1, found: 442.1. (Due to small scale, no 13C NMR or HRMS data were obtained.)
(Method A) Isolated as an orange oil in 96% yield: Rf=0.47 (orange spot, 7:3 hexanes:EtOAc); IR(cast film) 3359, 3055, 2950, 1746, 1705, 1608, 1460, 1241, 743 cm−1; 1H NMR (500 MHz, CDCl3) δ 8.29 (s, 1H), 7.84 (d, J=8.3 Hz, 1H), 7.67 (d, J=8.4 Hz, 1H), 7.64 (d, J=8.1 Hz, 1H), 7.48 (ddd, J=8.1, 6.8, 1.1 Hz, 1H), 7.42 (s, 1H), 7.29-7.23 (m, 6H), 7.17 (app t, J=8.1 Hz, 1H), 7.11 (dd, J=8.0, 1.5 Hz, 2H), 7.08 (app td, J=8.1, 7.2 Hz, 1H), 5.76 (br s, 1H), 5.27 (s, 2H), 3.81 (s, 3H); 13C NMR (125 MHz, CDCl3) δ 195.7, 169.3, 153.7, 139.9, 137.1, 136.9, 130.8, 129.6, 128.8, 128.6, 127.7, 127.6, 127.2, 126.9, 126.7, 126.2, 123.7, 122.4, 121.8, 120.2, 120.1, 110.8, 110.3, 107.0, 72.7, 53.7, 50.4; HRMS calc'd for C29H22N2O3Na [M+Na]+469.1523, found: 447.1530.
(Method A) Isolated as a yellow powder in 81% yield: m.p.=158-159° C.; Rf=0.19 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3368, 3055, 2919, 1732, 1711, 1635, 1491, 1225, 757 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.36 (br s, 1H), 7.69 (ddd, J=7.8, 1.3, 0.7 Hz, 1H), 7.57 (d, J=8.1 Hz, 1H), 7.52 (ddd, J=8.4, 7.1, 1.4 Hz, 1H), 7.32 (d, J=1.3 Hz, 1H), 7.30 (app t, J=1.0 Hz, 1H) 7.17 (ddd, J=8.3, 7.1, 1.3 Hz, 1H), 7.08 (ddd, J=8.2, 7.1, 1.2 Hz, 1H), 6.98 (dt, J=8.3, 0.7 Hz, 1H), 6.93 (ddd, J=7.9, 7.1, 0.9 Hz, 1H), 5.84 (tdd, J=5.6, 10.5, 17.2 Hz, 1H), 5.83 (br s, 1H), 5.23 (tdd, J=1.5, 1.5, 17.2 Hz, 1H), 5.16 (tdd, J=1.3, 1.3, 10.5 Hz, 1H), 4.71-4.67 (m, 2H); 13C NMR (125 MHz, CDCl3,): δ 194.8, 168.2, 161.1, 137.9, 136.5, 131.1, 125.4 (2×), 123.8, 122.6, 120.3 (2×), 119.8, 119.6, 119.2, 113.5, 111.7, 111.3, 72.6, 67.2; HRMS calc'd for C20H17N2O3 [M+H]+333.1234, found: 333.1232.
(Method A) Isolated as yellow crystals in 83% yield (using methanol as the recrystallization solvent): m.p.=148-150° C.; Rf=0.36 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3371, 3051, 2919, 1742, 1703, 1615, 1485, 1225, 743 cm−1; 1H NMR (500 MHz, CDCl3): δ 7.72 (d, J=7.8 Hz, 1H), 7.63 (d, J=8.1 Hz, 1H), 7.52 (ddd, J=8.3, 7.1, 1.3 Hz, 1H), 7.32-7.30 (m, 2H), 7.26 (ddd, J=8.2, 7.0, 1.1 Hz, 1H), 7.12 (ddd, J=8.0, 7.0, 1.1 Hz, 1H), 6.99 (d, J=8.2 Hz, 1H), 6.94 (app t, J=7.8 Hz, 1H), 5.88 (ddt, J=17.2, 10.5, 5.6 Hz, 1H), 5.83 (br s, 1H), 5.29 (ddt, J=17.2, 1.5, 1.5 Hz, 1H), 5.22 (ddt, J=10.4, 1.2, 1.2 Hz, 1H), 4.75-4.71 (m, 2H), 3.73 (s, 3H); 13C NMR (125 MHz, CDCl3,): δ 194.7, 168.3, 161.0, 137.8, 137.4, 131.2, 128.1, 126.0, 125.4, 122.2, 120.2, 119.9(2×), 119.8, 119.1, 113.5, 109.8, 109.7, 72.5, 67.1, 32.9; HRMS calc'd for C21H19N2O3 [M+H]+347.1390, found: 347.1393.
(Method A) Isolated as a yellow powder in 73% yield: m.p.=193-195° C.; Rf=0.21 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3351, 3011, 2915, 1738, 1712, 1635, 1465, 1233, 758 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.25 (br s, 1H), 7.70 (d, J=8.0 Hz, 1H), 7.60 (dd, J=8.1, 0.9 Hz, 1H), 7.54 (ddd, J=8.3, 7.2, 1.4 Hz, 1H), 7.43 (d, J=2.7 Hz, 1H), 7.37 (app dt, J=8.2, 0.8 Hz, 1H), 7.25 (dd, J=13.9, 6.2 Hz, 1H), 7.20 (ddd, J=8.2, 7.2, 1.1 Hz, 1H), 7.10 (ddd, J=8.2, 7.2, 1.0 Hz, 1H), 7.01 (d, J=8.3 Hz, 1H), 6.94 (app t, J=7.8 Hz, 1H), 5.68 (br s, 1H), 4.99 (dd, J=13.9, 2.0 Hz, 1H), 4.66 (dd, J=6.1, 2.0 Hz, 1H); 13C NMR (125 MHz, CDCl3): δ 194.0, 165.8, 160.9, 141.3, 138.0, 136.5, 125.5, 125.3, 123.7, 122.8, 120.5 (2×), 119.7, 119.6, 113.5, 111.6, 111.0, 100.0, 72.1; HRMS calc'd for C19H14N2O3Na [M+Na]+318.1004, found: 318.1007.
(Method A) Isolated as bright yellow crystals in 71% yield: m.p.=195-196° C.; Rf=0.20 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3364, 3052, 2952, 1711, 1610, 1242, 741 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.16 (m, 2H), 7.66 (d, J=8.5 Hz, 1H), 7.51 (ddd, J=8.4, 7.1, 1.4 Hz, 1H), 7.46-7.38 (m, 2H), 7.31-7.19 (m, 5H), 7.06 (ddd, J=8.1, 7.0, 1.0 Hz, 1H), 7.01 (d, J=8.3 Hz, 1H), 6.92 (app t, J=7.8 Hz, 1H), 6.26 (br s, 1H), 3.77 (s, 3H); 13C NMR (125 MHz, CDCl3) δ 195.2, 193.6, 161.1, 137.8, 137.4, 134.2, 133.2, 131.3, 127.9, 127.6, 126.1, 125.2, 122.4, 120.5, 120.3, 120.1, 119.3, 114.0, 112.0, 109.8, 79.0, 33.0; LC-MS calc'd for C24H19N2O2 [M+H]+366.1, found: 366.1.
(Method A) Isolated as bright yellow powder in 81% yield, inseparable mixture (HPLC, ca. 1.3:1 ratio of 2xa:2xb, characterized as mixture of diastereomers): Rf=0.21 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3362, 3051, 2955, 1705, 1618, 1242, 739 cm−1; 1H NMR (500 MHz, CDCl3): 8 (major isomer 2xa) 8.26 (br s, 1H), 7.69 (dddd, J=7.8, 1.4, 0.7, 0.7 Hz, 1H), 7.59-7.58 (m, 1H), 7.51 (ddd, J=8.3, 7.2, 1.4, 1H), 7.50 (ddd, J=8.3, 7.2, 1.4, 1H), 7.37 (d, J=2.7 Hz, 1H), 7.34 (ddd, J=8.2, 0.9, 0.9 Hz, 1H), 7.09 (ddd, J=8.1, 7.1, 1.1 Hz, 1H), 6.99 (dd, J=0.8, 0.8 Hz, 1H), 6.93 (ddd, J=7.1, 0.9, 0.9 Hz, 1H), 5.65 (br s, 1H) 4.77 (ddd, J=10.9, 10.9, 4.3 Hz, 1H), 0.84 (d, J=6.5 Hz, 3H), 0.82 (d, J=7.0 Hz, 3H), 0.64 (d, J=6.9 Hz, 3H), 8 (minor isomer 2xb) 8.24 (br s, 1H), 7.70 (dddd, J=7.8, 1.4, 0.6, 0.6 Hz, 1H), 7.61-7.60 (m, 1H), 7.38 (d, J=2.7 Hz, 1H), 7.33 (ddd, J=8.1, 0.9, 0.9 Hz, 1H), 7.06 (ddd, J=8.1, 7.1, 1.0 Hz, 1H), 7.00 (dd, J=0.7, 0.7 Hz, 1H), 6.92 (ddd, J=7.1, 0.9, 0.9 Hz, 1H), 5.76 (br s, 1H), 4.72 (ddd, J=11.0, 11.0, 4.4 Hz, 1H), 0.86 (d, J=6.5 Hz, 3H), 0.51 (d, J=7.1 Hz, 3H), 0.43 (d, J=6.9 Hz, 3H) (Note: some protons could not be properly assigned due to extensive spectral overlap); 13C NMR (125 MHz, CDCl3) δ 194.8, 194.6, 168.0 (2×), 161.2, 161.0, 137.7, 137.6, 136.6, 136.5, 125.7, 125.6, 125.4, 125.3, 123.7, 123.4, 122.6 (2×), 120.3, 120.2 (2×), 120.1 (3×), 120.0, 119.6, 113.7, 113.5, 111.9, 111.8, 111.5 (2×), 77.5, 77.4, 72.8, 72.7, 46.8, 46.7, 40.3, 40.2, 34.1 (2×), 31.5, 25.8, 25.5, 23.2, 23.0, 22.0, 20.8, 20.4, 15.9, 15.7 (Note: 2 aliphatic carbon resonances were not detected due to extensive spectral overlap); HRMS calc'd for C27H31N2O3 [M+H]+431.2329, found: 431.2332.
(Method A) Isolated as bright yellow powder in 79% yield, inseparable mixture (HPLC, ca. 2.3:1 ratio of 2ya:2yb, characterized as mixture of diastereomers): Rf=0.33 (yellow spot, 7:3 hexanes:EtOAc); IR (cast film) 3362, 3051, 2955, 1704, 1618, 1242, 740 cm−1; 1H NMR (500 MHz, CDCl3): 8 (major isomer 2ya) 7.58 (ddd, J=8.0, 1.0, 1.0 Hz, 1H), 7.51 (ddd, J=8.3, 7.1, 1.3 Hz, 1H), 7.06 (ddd, J=8.1, 7.1, 1.1 Hz), 5.65 (br s, 1H), 4.77 (ddd, J=10.8, 10.8, 4.3 Hz, 1H), 3.75 (s, 3H), 1.85 (sepd, J=7.1, 2.9 Hz, 1H), 0.87 (d, J=6.6 Hz, 3H), 0.83 (d, J=7.0 Hz, 3H), 0.64 (d, J=7.0 Hz, 3H), 8 (minor isomer 2yb) 7.59 (ddd, J=8.0, 0.9, 0.9 Hz, 1H), 7.50 (ddd, J=8.4, 7.2, 1.4, 1H), 7.09 (ddd, J=8.1, 7.1, 1.0 Hz, 1H), 5.76 (br s, 1H), 4.73 (ddd, J=10.9, 10.9, 4.4 Hz, 1H), 3.74 (s, 3H), 1.19 (sepd, J=7.0, 2.9 Hz), 0.86 (d, J=6.6 Hz, 3H), 0.52 (d, J=7.0 Hz, 3H), 0.44 (d, J=6.9 Hz, 3H) (Note: some protons could not be properly assigned due to extensive spectral overlap); 13C NMR (125 MHz, CDCl3) δ 194.9, 194.7, 168.1, 168.0, 161.2, 160.9, 137.7, 137.6, 137.3 (2×), 128.1, 127.9, 126.2, 126.0, 125.4, 125.3, 122.1 (2×), 120.2 (2×), 120.0 (3×), 119.7 (3×), 113.7, 113.5, 110.1, 110.0, 109.6, 109.5, 77.4, 77.3, 72.8, 72.7, 46.7, 46.6, 40.3, 40.2, 34.1 (2×), 32.9 (2×), 31.4, 25.7, 25.4, 23.1, 23.0, 21.9, 20.8, 20.4, 15.9, 15.7 (Note: two carbon resonances could not be found due to extensive overlap); HRMS calc'd for C28H32N2O3 [M+H]+445.2486, found: 445.2492.
Compounds 2za and 2zb:
(Method A) Isolated in 92% combined yield (2.7:1 ratio of 2za:2zb).
Major diastereomer 2za (isolated as green needles from partial separation of mixture): Rf=0.42 (yellow spot, 7:3 hexanes:EtOAc); [α]D20: +23.76 (c=1.01, DCM); IR (cast film) cm−1; 1H NMR (500 MHz, CDCl3): δ 7.68 (d, J=7.8 Hz, 1H), 7.62 (d, J=8.8 Hz, 1H), 7.52 (ddd, J=8.3, 7.2, 1.3, 1H), 7.39 (s, 1H), 7.30-7.22 (m, 4H), 7.14 (ddd, J=8.2, 7.1, 1.1, 1H), 7.13-7.11 (m, 2H), 7.06 (ddd, J=8.1, 7.1, 1.0, 1H), 7.01 (d, J=8.2 Hz, 1H), 6.92 (app t, J=7.8 Hz, 1H), 5.66 (br s, 1H), 5.28 (app br s, 2H), 4.75 (ddd, J=11.0, 11.0, 4.4 Hz, 1H), 1.91-1.88 (m, 1H), 1.81 (sepd, J=7.0, 2.9 Hz, 1H), 1.66-1.61 (m, 2H), 1.45-1.39 (m, 2H), 1.03-0.96 (m, 1H), 0.88 (app td, J=12.2, 11.0 Hz, 1H) 0.87-0.70 (m, 1H), 0.84 (d, J=6.6 Hz, 3H), 0.81 (d, J=7.0 Hz, 3H), 0.62 (d, J=7.0 Hz, 3H); 13C NMR (125 MHz, CDCl3) δ 194.8, 167.9, 161.0, 137.6, 137.1, 137.0, 128.8, 127.7, 127.6, 127.0, 126.3, 125.4, 122.3, 120.3, 120.2, 120.1, 120.0, 113.5, 110.7, 110.1, 77.4, 72.8, 50.3, 46.8, 40.2, 34.1, 31.4, 25.8, 23.2, 22.0, 20.7, 15.9; HRMS calc'd for C34H37N2O3 [M+H]+521.2799, found: 521.2810.
Minor diastereomer 2zb (partial assignment based on deduction, using spectra of mixture of diastereomers and spectra of major diastereomer; multiplicity of some peaks could not be properly assigned due to substantial overlap): 1H NMR (500 MHz, CDCl3): δ 7.68 (d, J=7.8 Hz, 1H), 7.65 (d, J=7.6 Hz, 1H), 7.51 (ddd, J=8.4, 7.2, 1.3 Hz, 1H), 7.41 (s, 1H), 7.30-7.22 (m, 4H), 7.15 (app t, J=7.2 Hz, 1H), 7.12-7.11 (m, 3H), 7.02 (d, J=8.16 Hz, 1H), 6.92 (app t, J=7.8 Hz, 1H), 5.80 (br s, 1H), 5.28 (app br s, 2H), 4.72 (ddd, J=11.0, 10.9, 4.4 Hz, 1H), 1.95-1.92 (m, 1H), 1.66-1.55 (m, 2H), 1.47-1.33 (m, 2H), 1.15 (sepd, J=6.9, 2.9 Hz, 1H), 0.87 (d, J=6.6 Hz, 3H), 0.48 (d, J=7.0 Hz, 3H), 0.40 (d, J=6.9 Hz, 3H) (NB.: three protons in aliphatic region could not be properly assigned due to extensive spectral overlap); 13C NMR (125 MHz, CDCl3) δ 194.5, 168.1, 161.2, 137.7, 137.2, 136.9, 128.8, 127.7, 127.4, 126.7, 126.5, 125.4, 122.3, 120.3, 120.2, 120.0, 113.7, 110.9, 110.1, 77.4, 72.7, 50.3, 46.7, 40.3, 34.1, 31.4, 25.5, 23.0, 22.0, 20.4, 15.7 (NB.: one resonance 13C was missing and could not be properly assigned due to extensive spectral overlap).
(Method A) Isolated as a bright yellow powder in 84% yield: m.p.=120° C. (dec.); Rf=0.32 (3:7, EtOAc:hexanes); IR(cast film) 3345, 3015, 2960, 1744, 1712, 1617, 1465, 1234, 733 cm−1 1H NMR (500 MHz, CDCl3): δ 8.41 (s, 1H), 7.50 (d, J=8.3 Hz, 1H), 7.40-7.28 (m, 6H), 7.21-7.16 (m, 2H), 7.05 (app. t, J=7.7 Hz, 1H), 6.91 (d, J=1.5 Hz, 1H), 6.89 (dd, J=8.3, 1.7 Hz, 1H), 5.73 (s, 1H), 3.22 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 193.7, 168.5, 160.8, 144.3, 138.0, 135.4, 132.4, 129.5, 128.8, 128.3, 126.7, 126.1, 122.6, 121.0, 120.6, 119.4, 118.4, 113.0, 111.3, 107.5, 73.6, 53.2; HRMS calc'd for C24H1735ClN2NaO3[M+Na]+439.0822, found 439.0822.
(Method A) Isolated as a bright yellow powder in 88% yield: m.p.=120° C. (dec.); Rf=0.17 (3:7, EtOAc:hexanes); IR (cast film) 3338, 3027, 2953, 1741, 1708, 1624, 1461, 1237, 747 cm−11H NMR (500 MHz, CDCl3): δ 8.20 (s, 1H), 7.52 (d, J=8.2 Hz, 1H), 7.33-7.30 (m, 3H), 7.19-7.15 (m, 2H), 7.03 (ddd, J=8.2, 7.2, 1.1 Hz, 1H), 6.94 (dd, J=1.7, 0.6 Hz, 1H), 6.89-6.85 (m, 3H), 5.63 (s, 1H), 3.82 (s, 3H), 3.31 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 193.7, 168.6, 160.8, 160.1, 144.3, 137.9, 135.3, 130.9, 126.8, 126.2, 124.7, 122.6, 121.0, 120.6, 119.4, 118.6, 113.8, 113.1, 111.2, 107.5, 73.6, 55.4; 53.4; HRMS calc'd for C25H2035ClN2O4[M+H]+447.1106, found 447.1105.
(Method A) Isolated as a bright yellow powder in 67% yield: m.p.=135° C. (dec.), Rf=0.21 (3:7, EtOAc:hexanes); IR(cast film) 3336, 3021, 2945, 1739, 1712, 1615, 1471, 1239, 749 cm−1 1H NMR (500 MHz, CDCl3): δ 8.15 (s, 1H), 7.51 (dd, J=8.2, 0.6 Hz, 1H), 7.43-7.40 (m, 2H), 7.36 (d, J=8.0 Hz, 1H), 7.22-7.18 (m, 2H), 7.07-7.04 (m, 3H), 6.98 (dd, J=1.7, 0.5 Hz, 1H), 6.90 (dd, J=8.2, 1.7 Hz, 1H), 5.62 (s, 1H), 3.35 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 193.4, 168.5, 163.1 (d, J=249.5 Hz), 160.7, 144.5, 136.8, 135.4, 131.6 (d, J=8.3 Hz), 128.5, 126.7, 126.2, 123.0, 121.3, 120.9, 119.4, 118.6, 115.5 (d, J=21.7 Hz), 113.1, 111.2, 108.2, 73.4, 53.5; 19F NMR (469 MHz, CDCl3): δ-111.7 (br s); HRMS calc'd for C24H1735ClFN2O3 [M+H]+435.0904, found 435.0906.
(Method A) Isolated as a bright yellow solid in 51% yield: m.p.=148° C. (dec.), Rf=0.30 (3:7, EtOAc:hexanes); IR(cast film) 3344, 3039, 2957, 1729, 1712, 1659, 1464, 1239, 731 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.17 (br s, 1H), 7.48 (d, J=8.2 Hz, 1H), 7.47 (dd, J=7.0, 2.2 Hz, 1H), 7.37 (d, J=10.3 Hz, 1H), 7.32 (ddd, J=6.8, 4.6, 2.2 Hz, 1H), 7.27-7.21 (m, 2H), 7.13-7.07 (m, 2H), 7.01 (d, J=1.6 Hz, 1H), 6.91 (dd, J=8.3, 1.7 Hz, 1H), 5.66 (s, 1H), 3.48 (s, 3H); 13C NMR (125 MHz, CDCl3): 6 193.2, 168.6, 163.0 (d, J=249.5 Hz), 160.4, 157.4, 144.6, 135.5, 135.2, 132.0, 129.5 (d, J=19.9 Hz), 129.4, 126.8, 126.1, 123.3, 121.5, 121.0, 119.3, 118.7, 116.6 (d, J=25.1 Hz), 113.2, 111.4, 108.9, 73.2, 53.6; 19F NMR (469 MHz, CDCl3): δ-114.2 (br s); HRMS calc'd for C24H15Cl2FN2O3Na [M+Na]+491.0336, found 491.0334.
(Method A) Isolated as a bright yellow powder in 74% yield; m.p.=120° C. (dec.); Rf=0.32 (3:7, EtOAc:hexanes); IR(cast film) 3342, 3025, 2950, 1741, 1709, 1618, 1460, 1231, 733 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.32 (s, 1H), 7.42 (d, J=8.3 Hz, 1H), 7.26-7.39 (m, 6H), 7.14-7.19 (m, 2H), 7.10 (d, J=1.5 Hz, 1H), 7.01-7.05 (m, 2H), 5.67 (s, 1H), 3.21 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 193.9, 168.4, 160.8, 138.0, 135.4, 133.3, 132.4, 129.5, 128.8, 128.4, 126.7, 126.2, 123.8 122.7, 120.6, 119.4, 118.8, 116.1, 111.3, 107.6, 73.5, 53.2. HRMS calc'd for C24H18BrN2O3[M+H]+461.0495, found 461.0501.
(Method A) Isolated as a bright yellow solid in 76% yield; m.p.=120° C. (dec.); Rf=0.32 (3:7, EtOAc:hexanes); IR(cast film) 3347, 3033, 2956, 1731, 1713, 1638, 1465, 1236, 713 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.12 (br s, 1H), 7.46 (d, J=8.3 Hz, 1H), 7.26-7.12 (m, 3H), 7.19-7.14 (m, 3H), 7.05-7.03 (m, 2H), 7.02-6.88 (m, 2H), 5.59 (s, 1H), 3.83 (s, 3H), 3.32 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 193.9, 168.6, 160.8, 160.1, 138.0, 135.4, 133.3, 130.9, 126.8, 126.2, 124.7, 123.8, 121.0, 120.6, 119.4, 118.7, 116.1, 113.8, 111.2, 107.6, 73.5, 55.4, 53.4; FIRMS calc'd for C25H2079BrN2O4 [M+H]+491.0601, found 491.0605.
(Method A) Isolated as a bright yellow solid in 52% yield: m.p.=120° C. (dec.); Rf=0.32 (3:7, EtOAc:hexanes); IR(cast film) 3339, 3035, 2954, 1728, 1718, 1648, 1464, 1229, 728 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.17 (br s, 1H), 7.46 (d, J=8.3 Hz, 1H), 7.26-7.12 (m, 3H), 7.19-7.14 (m, 3H), 7.05-7.03 (m, 2H), 7.02-6.88 (m, 2H), 5.59 (s, 1H), 3.32 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 193.7, 168.4, 163.1 (d, J=249.5 Hz), 160.7, 136.8, 135.4, 133.4, 131.6 (d, J=8.3 Hz), 128.5, 126.7, 126.2, 124.0, 123.0, 120.9, 119.4, 119.0, 116.2, 115.5 (d, J=21.6 Hz), 111.2, 108.1, 73.3, 53.5;19F NMR (469 MHz, CDCl3): δ-111.7 (br s); HRMS calc'd for C24H1779BrFN2O3 [M+H]479.0402, found 479.0404.
(Method A) Isolated as a bright yellow solid in 38% yield: m.p.=120° C. (dec.); Rf=0.30 (3:7, EtOAc:hexanes); IR(cast film) 3341, 3037, 2951, 1724, 1716, 1651, 1465, 1234, 737 cm−1; 1H NMR (500 MHz, CDCl3): δ 8.21 (br s, 1H), 7.46 (dd, J=7.0, 2.2 Hz, 1H), 7.40 (d, J=8.2 Hz, 1H), 7.35 (d, J=8.2 Hz, 1H), 7.30 (ddd, J=6.8, 4.5, 2.2 Hz, 1H), 7.24-7.20 (m, 2H), 7.19 (dd, J=1.5, 0.4 Hz, 1H), 7.11-7.06 (m, 3H), 5.66 (s, 1H), 3.48 (s, 3H); 13C NMR (125 MHz, CDCl3): δ 193.2, 168.6, 163.0 (d, J=249.5 Hz), 160.4, 151.1, 142.0, 133.3, 133.2, 132.0, 129.5, 129.4 (d, J=19.9 Hz), 126.8, 126.1, 124.5, 123.2, 121.0, 119.3, 119.0, 116.6 (d, J=25.1 Hz), 116.3, 111.4, 108.9, 73.2, 53.6; 19F NMR (469 MHz, CDCl3): δ-114.2 (br s); HRMS calc'd for C24H1679Br35ClFN2O3[M+H]+513.0012, found 513.0015.
Virology Procedures
Cells and virus. Hep-2 cells were grown in Opti-MEM media supplemented with 2% fetal bovine serum (FBS), 1HAEo-cells were grown in MEM media supplemented with 10% FBS, and Hela cells grown in DMEM supplemented with 10% FBS. Infections were conducted using human RSV strain A2 or RSV-A2-GFP strain described previously [Hallak, L. K.; Spillmann, D.; Collins, P. L.; Peeples, M. E. J. Virol. 2000, 74, 10508-10513]. A549 cells and Vero cells were grown in DMEM supplemented with 10% FBS and used for experiments with ZIKV (Cambodia strain).
Structure activity relationship screens of antiviral activity. 1HAEo-cells were seeded at 80% confluency and incubated overnight. In SAR1 cells were preincubated with compounds for one hour at 20 μM prior to infection, followed by dilution to 10 μM and infection with RSV-A2-GFP at an MOI of 1 for 2 hours. In SAR2 1HAEo-cells were infected with RSV-A2-GFP at an MOI of 0.5 for 2 hours without pre-incubation with compounds. At 2 hours post infection, infectious media was removed from wells to prevent carry over of infecting virus into progeny collections. Fresh media containing compounds at 10 μM was added for remainder of 48 hour incubation. HeLa cells, used to titrate progeny virus, were plated at 80% confluency and incubated overnight. Progeny virus collected at 48 hours was immediately transferred to HeLa cells in duplicate technical replicates without dilution. At 2 hours post infection media was replaced with fresh compound-free media. Infection was stopped at 20 hours post infection, preventing cell to cell spread of virus, and infected cells were identified via indirect immunofluorescence assay.
Progeny virus quantification via indirect immunofluorescence. HeLa cells were fixed in 1:1 methanol:acetone, incubated with goat anti-RSV polyclonal antibody (Meridian Life Science B65860G) for 1 hour at room temperature, and then incubated with chicken anti-goat polyclonal antibody conjugated to Alexafluor 647 (LifeTech A-21469). Finally, cell nuclei were stained with DAPI. Total cells (identified by DAPI staining) and infected cells were counted via the Operetta high content imaging system at 20x objective magnification.
MTT cell viability assay. 1HAEo-cells were grown in parallel with infected and compound treated cells in SAR and dose response experiments. 0.3 mg/mL of MTT ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)) in PBS was added to wells and incubated for 1 hour. Precipitate was dissolved in DMSO and absorbance was measured at 570 nm.
Cell extract-based in vitro inhibition of viral transcription. The protocol utilized to examine in vitro viral transcription was first developed for virus vesicular stomatitis virus [Canter, D. M.; Jackson, R. L.; Perrault, J. Virology 1993, 194, 518-529], and adapted for RSV as published previously [Noton, S. L.; Nagendra, K.; Dunn, E. F.; Mawhorter, M. E.; Yu, Q.; Fearns, R. J. Virol. 2015, 89, 7786-7798]. Briefly, HEp-2 cells were grown in 6 well plates overnight to approximately 80% confluence and infected with RSV A2 at an MOI of 5 for 1 hour before incubating overnight in fresh media. Immediately before cell lysis, cells were incubated in 2 μg/mL actinomycin D to block cellular transcription. Cells were lysed on ice via 1 minute treatment with lysolecithin, and cell extract was subsequently collected in transcription buffer containing 50 mM Tris-acetate at pH 8, 8 mM Mg acetate, 300 mM K acetate, 2 mM DTT, 1 mM spermidine, 10 mM creatine phosphatase, 1 μg/mL aprotinin, 16 U creatine phosphokinase, 1 mM each of ATP, GTP, and CTP, 50 μM UTP, and 2 μg/mL actinomycin D. Cell debris was removed via refrigerated centrifugation. In vitro transcription reaction was performed by combining soluble cell extract, additional transcription buffer, RNase Inhibitor, compound 28 diluted in DMSO, and 10 μCi [α 32-P] UTP, incubated at 30° C. for 3 hours. RNA extraction was performed with Qiagen RNeasy kit as per manufacturer protocol to purify samples, followed by hybridization with oligo(dT) to facilitate RNaseH digestion of mRNA transcript poly(A) tails. Denaturing electrophoresis of samples was performed using 4% acrylamide containing 7 M urea gels. Gels were dried and autoradiography captured during 4 day exposure. Viral RNA products were quantified by densitometry analysis following phosphorimaging (n=4).
Dose response experiments. 1HAEo-cells were infected as described, progeny virus collected, and tittered on HeLa cells in duplicate as described in SAR 2. During initial infection in 1HAEo-cells infectious media was replaced with media containing compounds at indicated concentrations at 2 hours post infection. Proportion of virus infected cells were quantified via indirect immunofluorescence assay.
Mutant escape assay. 1HAEo-cells were infected with RSV-GFP-A2 at an initial MOI of 0.1 and serial passaged over incubation periods of 48-72 hours. Samples were collected during passaging and stored in liquid nitrogen for tittering via indirect immunofluorescence assay. Ongoing low level viral infection during passaging was confirmed by identification of green fluorescence from GFP produced by the RSV-GFP in infected cells.
Zika Virus antiviral screen. Vero cells were pre-treated with bis(indole) compounds at 10 μM for 4 hours before infection with ZIKV at an MOI of 1. After 48 hour infection progeny ZIKV was collected, serial diluted over 6 logs, and transferred onto fresh Vero cell monolayers and incubated for 4 days. Vero cell monolayers were then fixed with 10% formaldehyde and stained with crystal violet; viral infection was quantified via plaque assay.
Results and Discussion
Conditions originally employed for the cascade process utilized 10 mol % Cu(hfacac)2 and heating (toluene at reflux). Catalytic generation of copper metallocarbenes from diazoketone precursors is generally accepted to require the Cu(I) oxidation state, necessitating in situ reduction of the starting Cu(II) salt by added reductant or a sacrificial quantity of the diazo substrate. Once tthe C-acylimine was formed, it was considered that nucleophilic trapping involved activation of the imine nitrogen atom by either Cu(I) or unreduced Cu(II). Using substrate 1a and indole as the trap, only moderate yields of the desired adduct 2a were observed under standard conditions, complicated by varying amounts of the regioisomer resulting from Friedel-Crafts trapping at C-2 (Table 1, entry 1). Attempts to improve selectivity by reducing reaction temperature were not fruitful, providing only traces of 2a after 24 h, which was attributed to slow production of catalytically competent Cu(I) (entry 2). Cu(OTf).PhCH3 in DCM at rt afforded 2a in only 12% yield, although 1a was consumed quickly (Table 1, entry 3). In contrast, use of 10 mol % Cu(OTf)2 required extended stirring in DCM at rt, but ultimately provided 2a in 92% yield (Table 1, entry 4), and these conditions were scalable (Table 1, entry 5). Induction time was consistent with a pre-activation step to generate CuOTf, but the high yield in this case (in contrast to entry 3) suggested a requirement for another component besides Cu(I) to attain efficient production of indole adduct 2a.
Species responsible for reduction of Cu(OTf)2 was not clear. As noted above, diazo compounds are known to play this role, but the presence of excess indole suggested an alternative possibility. Gaunt and co-workers have shown that direct C-3 arylation of indoles can be accomplished using catalytic Cu(OTf)2, involving C-3 cupration of the indole by a Cu(III) intermediate, ArCu(OTf)2, which is formed via CuOTf insertion to an aryliodonium salt [Phipps, R. J.; Grimster, N. P.; Gaunt, M. J. J. Am. Chem. Soc. 2008, 130, 8172-8174]. In the present context, it was considered that indole C-3 cupration by Cu(OTf)2 could form organocopper(II) complex 3, which could furnish CuOTf through hemolytic cleavage of the weak C—Cu bond (See
One equivalent of the strong Bronsted acid TfOH was produced along with each equivalent of CuOTf, and its presence appeared to be required for clean trapping by indole to furnish 2a. Thus, while treatment of 1a with 10 mol % CuOTf.PhMe gave minor amounts of 2a despite complete consumption of starting material (Table 1, entry 3), when 10 mol % Cu(OTf)2 was pretreated with 0.2 equiv indole and stirred for 2 h (green solution) then added to a solution of 1a and 1.8 equiv indole, 2a was obtained in 83% yield. The exact nature of the Bronsted acid catalyst responsible for indole addition was unclear, but the indolylindoline triflate salt 4 was considered a feasible candidate. This dimeric product is readily formed from indole in the presence of Bronsted or Lewis acid and various electrophilic reagents, and evidence for its formation was observed during the indole/copper(II) redox process (see Example 2). Overall, the cascade process was considered to involve a dual catalytic cycle involving Cu(I)-catalyzed conversion of 1a to the cyclized indolen-3-one intermediate, followed by Bronsted acid activation of the imine (and concomitant turnover of Cu(I)) for Friedel-Crafts alkylation of indole to afford 2a (see
Direct involvement of Bronsted acid in the indole addition step is borne out by the observation of an enantiomeric excess of 36% when the reaction was carried out in the presence of (+)-camphorsulfonic acid (see
To test the scope of the dual catalytic process, other indole derivatives with varying steric and electronic properties were subjected to the optimal reaction conditions (see
Effects of substitution on the diazoazide partner were also examined. Substitution with an electron donating methyl or methoxy group gave better yields of adducts 21-n (84-94%) than the electron-withdrawing nitro group (2o, 14% along with 68-76% recovered starting material). The presence of halogen substituent (chloro) meta to the azide or extending the aromatic system were tolerated, furnishing adducts 2p-s in good yields. Allyl, vinyl, or menthyl esters adjacent to the diazo group were tolerated, as was phenyl ketone, affording 2t-2z in good to excellent yields.
With rapid access to a library of truncated isatasine A analogues available, their potential antiviral activity against RSV was investigated. An initial library consisting of a select group of the compounds depicted in
While numerous fusion inhibitors for RSV have been identified, including the prophylactic monoclonal antibody Palivizumab, they have yet to translate into therapy for the treatment of established RSV infections. This may be due to the nature of Palivizumab targeting only the entry step of the viral replication cycle. Therefore inhibiting the RSV replication complex after RSV has entered a cell was considered a desirable mechanism of action to block the spread of existing infection. To elucidate a mechanism of antiviral activity, root antiviral compound 5a was assayed for activity against the RSV viral replication complex that serves as a marker of RSV replication in the host cell. Decreasing viral transcription from the RSV replication complex was observed in a dose dependent manner in the presence of compound 5a in vitro (see
To further assess the therapeutic potential of compounds identified in SAR 1 and SAR 2, a therapeutic index (TI) of each compound was identified (e.g., ratio of effective concentration at which viral replication is reduced by half (EC50) to cytotoxic concentration at which cell viability is reduced by half (CC50)). Compound 2p was included as a cytotoxicity control, having demonstrated cytotoxicity in SAR 1. Cell viability decreased rapidly with increasing concentration of compound 2p, with a CC50 value of 22.8 μM, while viral replication was not diminished at compound concentrations tolerated by the host cell. In contrast, compound 5a and the 5 series of compounds derived from 5a consistently inhibited viral replication at concentrations tolerated by the host cell. In SAR 2, addition of OMe at R1 decreased the toxicity of these compounds, with CC50 values increasing from 42.5 μM for compound 5a, to 59.2 μM and 93.5 μM for compounds 5b and 5f respectively. Overall, the TI was increased from 4.1 for compound 5a to 12.3 with compound 5f. These increases in TI suggested that the compounds were capable of interfering with viral replication at concentrations tolerated by the host cell.
An established challenge to development of antiviral pharmaceuticals is the evolution of antiviral resistance. This is pronounced for viruses with RNA genome due to an inherent error-prone process of RNA-dependent RNA polymerase replication of a viral genome. Error prone replication results in a high mutation rate of RNA viruses that leads to development of antiviral resistance. As such, development of resistant mutants to compounds 5a and 5f was undertaken, as the site of mutations could provide further insights into the nature of the viral target. However, despite development of resistance against other transcription inhibitors in 6-8 passages, 14 serial passages over five weeks did not result in the development of viruses resistant to either compound (see
ZIKV constitutes a virus distinct from RSV with a markedly different intracellular replication strategy. Both viruses encode a viral RdRp essential to viral replication; however, the structure and function of RSV RdRp and ZIKV RdRp are quite different. RSV is a Pneumoviridae family virus with a negative sense RNA genome, and hence RSV RdRp must be packaged with an infecting virion for transcription to occur. RSV polymerase recognizes cis-acting ‘gene start’ and ‘gene end’ elements of the viral genome to produce subgenomic mRNAs enabling translation of viral proteins. In contrast, ZIKV is a Flaviviridae family virus having a positive sense RNA genome. Therefore, the ZIKV genome can be directly translated into one large viral polyprotein by host ribosomes without a need for production of viral subgenomic mRNAs. Thus the primary function of ZIKV RdRp is to facilitate genome replication through a negative sense antigenome intermediate. As RSV and ZIKV represent taxonomically distinct viruses, antiviral activity of the isatisine A-inspired bis(indole) compounds were examined against ZIKV to determine whether their antiviral activities may be broadly acting. ZIKV infection was screened in the presence of compounds 2d, 2o, 5a, and 5a derivatives (5b-5h) in VERO cells. Significant inhibition of ZIKV by compound 5a and all 5a derivatives was observed (see
As such, a dual catalytic cascade process was discovered that rapidly assembles drug-like bis(indole) scaffolds under mild, convenient conditions whose only by-products are 2 equiv of N2. Indole trapping agents served to activate a Cu(OTf)2 precatalyst, while also producing a Bronsted acid catalyst required for Friedel-Crafts alkylation of indoles by the intermediate C-acylimines. Preliminary experiments demonstrated that added chiral acids afforded moderate enantioselectivity. Some of the bis(indole) products showed promising antiviral activity against important pathogens RSV and ZIKV. The modular nature of the synthetic transformation may allow for the development of improved analogues against these and other undesirable pathogens.
aProduct isolated as a mixture of regioisomers.
b1a was rapidly consumed with formation of multiple colored products.
cGram-scale reaction with direct recrystallization of crude product.
dMultiple colored products were observed.
eL* = bis(oxazoline) ligands shown above.
Compound Stability Studies
Differential Scanning calorimetry (DSC): An example of a DSC experiment for compound 1a is depicted in
Thermogravimetric analysis (TGA): The decomposition pattern was further monitored using a TGA experiment. An example of a TGA experiment for compound 1a is shown in
Both of these experiments shed some light on the thermal stability of 1a. These experiments provided evidence that the metal catalyzed transformation, associated with the starting material 1a, at ambient temperature (22° C. to 40° C.), described in the next section (vide infra), was not merely heat driven as the starting material 1a was stable at this temperature range. Moreover, using the DSC data, it was anticipated that there would be less risk of detonation, due to thermal runaway, associated with compound 1a, or analogous diazoazides, provided the material was stored at freezer temperature (<0° C.) when not in use. Long-term storage (>1 week) is generally not advisable for compound 1a, especially for a large-scale preparation, but it was noticed that compound 1a and the other analogues survived with little or no decomposition during storage at freezer temperature (−20° C.) for >3 months (monitored weekly using 1H NMR).
These experiments (DSC and TGA) alone should not, however, be taken as an indication that the material is completely safe for handling. Other parameters, for example potential shock sensitivity, are equally important. Attempts to carry out a hammer test for compound 1a (ca. 0.5 mg) to evaluate its shock sensitivity were inconclusive. A small spark was observed, but in the absence of a reliable benchmark, it was not possible to draw any conclusions regarding the shock sensitivity of 1a. Similar to other common potentially explosive materials used in organic synthesis laboratories, such as peroxides, it is cautioned to potential users to treat the starting material 1a and analogues with due care and respect. Any unanticipated detonation of these diazo azide starting materials was not observed, and most large-scale reactions involving these substrates were performed in a well-ventilated fume hood equipped with a blast shield. It is recommended to wear a Kevlar® apron and safety gloves and using earplugs as added safety precautions, especially for large-scale reactions.
Mechanistic Considerations
Copper Oxidation State in the Catalytic Cycle
Conversion Studies (% Recovery):
Copper (II) oxidation state:
A solution of diazo-azide 1a (50 mg, 0.20 mmol) in DCM (5 mL) was added to a solution of Cu(OTf)2 (7.3 mg, 0.020 mmol) in DCM (5 mL) at room temperature via syringe pump over 1 h. Once addition was complete, the reaction was monitored by TLC for consumption of the diazo-azide starting material. After 24 h, the solution was extracted with water (10 mL, 3×) to remove the copper salt, and the DCM solution was dried with MgSO4, filtered and concentrated under reduced pressure. The crude oil was purified using silica gel flash column chromatography eluting with 20% EtOAc/Hexanes. This procedure resulted in recovery of diazo-azide 1a, in 81-93% (three repeats). Using identical conditions but allowing the mixture to stir for 5 d instead of 24 h resulted in recovery of 73-84% (three repeats) starting material 1a. Analysis of TLC and NMR spectra of the crude mixture revealed the presence of starting material, and a faint spot on the baseline using 20% EtOAc/Hexanes as the eluent (TLC).
Copper (I) Oxidation State:
A solution of diazo-azide 1a (50 mg, 0.20 mmol) in DCM (5 mL) was added to a solution of Cu(OTf)(PhMe) (10 mg, 0.020 mmol) in DCM (5 mL) at room temperature via syringe pump over 1 h. Once addition was complete, the reaction was monitored by TLC for consumption of the diazo-azide starting material. After 24 h, the solution was extracted with water (10 mL, 3×) to remove the copper salt, the DCM solution was dried with MgSO4, filtered and concentrated under reduced pressure. The crude oil was purified using silica gel flash column chromatography eluting with 20% EtOAc/Hexanes. This procedure resulted in recovery of trace amount of 1a (<5%). Analysis of a TLC chromatogram of the crude mixture revealed the presence of multiple colored spots. A crude NMR spectra revealed an intractable mixture of multiple compounds.
Conversion Studies (in-Situ IR Analysis and 1H-NMR Spectroscopy)
Copper (II) Oxidation State:
Using IR Spectroscopy:
A solution of diazo-azide 1a (50 mg, 0.20 mmol) in DCM (5 mL) was added to a solution of Cu(OTf)2 (7.6 mg, 0.02 mmol) in DCM (5 mL) at room temperature via syringe pump over 1 h. Once addition was complete, the reaction was monitored by TLC for consumption of the diazo-azide starting material. An aliquot of the stirred solution (0.5 mL), was taken after ca. 5 min, 1 h, 2 h, 3 h, and 24 h of stirring, each aliquot was diluted with DCM (1 mL). The diluted solution was analyzed using IR spectroscopy and plotted as overlaid spectra (
Copper (I) Oxidation State:
A solution of diazo-azide 1a (50 mg, 0.20 mmol) in DCM (5 mL) was added to a solution of Cu(OTf)(PhMe) (10 mg, 0.020 mmol) in DCM (5 mL) at room temperature via syringe pump over 1 h. Once addition was complete, the reaction was monitored by TLC for consumption of the diazo-azide starting material. An aliquot of the stirred solution (0.5 mL), was taken after ca. 5 min, 1 h, 2 h, 3 h and 24 h of stirring, each aliquot was diluted with DCM (1 mL). The diluted solution was analyzed using IR spectroscopy and plotted as overlaid spectra (
A complete absence of a broad peak at 2137 cm−1 indicated complete decomposition of the diazoketone and the azide functional groups. An increase in the absorbance and broadening of signal around 1730 cm−1 was noticed on diazo-azide starting material upon exposure with copper (II) triflate over 24 h, indicative of carbonyl interaction (dative bond) with copper (II), which did not progress to substantial diazo-ketone decomposition (notice continued substantial absorbance at 2137 cm-1). However, a notable decrease in the absorbance of the signal around 2137 cm−1 was noticed on diazo-azide starting material upon exposure with copper (I) triflate over 24 h, indicative of complete decomposition of diazoketone and azide over a 24 h period. This time period is within the observed optimized reaction time (16-36 h, depending on substitution of indole substrate) for azide-metallocarbene coupling, followed by Friedel-Crafts alkylation with indole. This pair of experiments, together with the recovery analysis, and 1H-NMR spectroscopy (vide infra) provided a distinction between the catalytic behaviors of the two copper oxidation states.
Using 1H-NMR Spectroscopy:
In a 2.0 mL vial, a solution of diazo-azide 1a (10 mg, 0.040 mmol) in deuterated DCM (0.5 mL) at room temperature was added to Cu(OTf)(PhMe) (ca. 2.0 mg, 0.0039 mmol) as a solid. The solution was quickly transferred in an NMR tube, capped with septum, and purged with argon. NMR spectra (400 MHz) were acquired once per hour over a 10 h period. The array of spectra was plotted. After 8-10 h, the starting material 1a was completely consumed. These data, corroborated with the IR results, provided evidence that copper (I) was the kinetically competent oxidation state during conversion of diazoazide into C-acylimine (see
After extensive analysis of the copper oxidation state necessary for decomposition of starting material 1a, it was posited that copper (I) was the kinetically competent oxidation state. This, however, was not expected since the initially loaded catalyst was copper (II) triflate.
Formation of Active Copper (I) Catalyst from Copper (II) Precatalyst
Using UV Spectroscopy, Formation of a New σ-Copper Indole Complex:
Solution A: A solution of indole (55 mg, 0.20 mmol) was added to Cu(OTf)2 (7.3 mg, 0.020 mmol) in DCM (5 mL) at room temperature. Once addition was complete, formation of light green solution occurred after 1 h. An aliquot of this solution (1 mL) was diluted with 5 mL of DCM and was subjected to UV spectroscopy. Solution B: An aliquot (1 mL) of a solution of indole dissolved in DCM (0.02 M, 10 mL) at room temperature was also subjected to UV spectroscopy. Solution C: An aliquot (1 mL) of a solution of Cu(OTf)2 dissolved in DCM (0.02 M, 10 mL) at room temperature was also subjected to UV spectroscopy. Further dilution was necessary until peaks (>300 nm) could be seen. After analysis of the three spectra, formation a broad new peak at 395 nm was seen from Solution A, indicating formation of a new colored complex (
Using ESI-MS, Detection of the Mass Fragments Corresponding to the Ddimer and the Copper-Indole Complex:
A solution of indole (55 mg, 0.20 mmol) was added to Cu(OTf)2 (7.3 mg, 0.020 mmol) in DCM (5 mL) at room temperature. Once addition was done, formation of light green solution occurred after one hour. An aliquot of this solution (1 mL) was diluted with acetonitrile and directly subjected to ESI-MS analysis. After analysis of the ESI-MS (
Given an apparent reaction between the indole and copper (II) triflate to afford an initially colored complex, the absorption detected through UV-VIS spectroscopy, and literature precedent [Rauniyar, V.; Wang, Z. J.; Burks, H. E.; Toste, F. D. J. Am. Chem. Soc. 2011, 133, 8486-8489], it was posited reduction of copper (II) triflate through the initially formed metallated indole, followed by disproportionation reaction. This disproportation reaction could theoretically generate copper (I) triflate, and copper (III) triflate indole complex. The latter could reductively eliminate to copper (I) triflate and indole triflate. Based on these experiments, the kinetically competent copper (I) triflate was proposed to result from a redox process involving copper (II) precatalyst and indole. However, this proposal on its own could not explain the substantial difference in isolated yield of 2a between Cu(OTf)2 and CuOTf. An additional component in the reaction, presumably derived from the activation reaction of Cu(OTf)2 and indole, was necessary for efficient Friedel Crafts alkylation with indole. It was proposed that a Bronsted acid catalyst, either TfOH or its salt with indole dimer B, acted to catalyze Friedel-Crafts addition of indole to the intermediate C-acylimine formed from metallocarbene-azide coupling.
Evidence for Bronsted Acid Catalysis of Friedel-Crafts Alkylation
NOTE: Since the putative C-acylimine occuring in the Cu(I)-catalyzed coupling step was transient and could not be isolated, isolable C-acylimine 6 was used as a model to help understand the second catalytic cycle.
Stirring C-acylimine 6 with Indole in the Presence of Cu(OTf)(PhMe):
A solution of C-acylimine 6 (50 mg, 0.19 mmol) and indole (50 mg, 0.20 mmol) was added to Cu(OTf)(PhMe) (9.7 mg, ca 0.020 mmol) in DCM (5 mL) at room temperature. Once addition was complete, the solution was allowed to stir overnight. After stirring for 16 h, the solution was extracted with water (5 mL, 2×), and the organic layer was dried with MgSO4, filtered and concentrated under reduced pressure. Analysis of crude reaction mixture indicated that substantial quantities of 6 were present. Upon purification via a short pad of silica (20% EtOAc/Hexanes), ca. 91% of 6 was recovered. This model reaction showed that, although copper (I) triflate could convert 1a to the C-acylimine (see previous experiments), copper (I) could not efficiently catalyze the Friedel-Crafts alkylation reaction with indole.
Stirring C-Acylimine 6 with Indole in the Presence of TfOH:
A solution of C-acylimine 6 (50 mg, 0.19 mmol) and indole (50 mg, 0.20 mmol) was mixed with excess TfOH (ca. 50 μL, ca. 0.57 mmol) in DCM (5 mL) at room temperature. Once mixing was complete, within 5 minutes a change in color was noticed, from deep purple to light orange. The solution was diluted with water (5 mL) and extracted with DCM (5 mL, 3×). The organic layer was dried using MgSO4 concentrated under reduced pressure. Analysis of the crude mixture indicated that 6 was completely consumed, and adduct 7 can be detected using crude NMR. Purification of 7, however, was hampered by the presence of several side products.
Stirring C-Acylimine 6 with Indole in the Presence of CSA:
A solution of C-acylimine 6 (50 mg, 0.19 mmol) and indole (50 mg, 0.20 mmol) was mixed with camphorsulfonic acid CSA (4.0 mg, ca. 0.020 mmol) in DCM (5 mL) at room temperature. Once addition was complete, the solution gradually (overnight) changed color from deep purple to a yellow suspension. Analysis of the TLC indicated that 6 was completely consumed. The suspension was filtered. The filtered solid was dissolved in deuterated DMSO and was analyzed using NMR spectroscopy. Analysis of the spectra revealed that the 2:1 adduct, 7, (indole:indol-3-one), along with water and DMSO, were the only detectable components present in solution.
These results indicates that Bronsted acid, produced during the reduction of copper (II) triflate by indole, could activate the C-acylimine towards Friedel-Crafts alkylation reaction.
Using a (+)-Camphorsulfonic Acid as a Chiral Co-Catalyst:
A solution of diazo-azide 1a (50 mg, 0.20 mmol) in DCM (5 mL) was added to a solution of indole (46 mg, 0.39 mmol), Cu(OTf)2 (7.3 mg, 0.020 mmol), and the (+)-CSA (4.0 mg, ca. 0.02 mmol) in DCM (5 mL) at room temperature via syringe pump over 1 h. The reaction mixture turned light green over 2 h, then slowly turned dark brown over 24 h. Once addition was complete, the reaction was monitored by TLC for consumption of 1a. Upon consumption of 1a, the reaction mixture was poured in an Erlenmeyer flask, thoroughly dissolved in ethyl acetate (ca 10 mL), dried over MgSO4, filtered, concentrated under reduced pressure and purified by flash chromatography. (NB.: crude mixture after concentration was insoluble in DCM. In this case, necessarily, ethyl acetate was used as a solvent to load the sample onto silica gel (silica gel, 7:3 hexanes:EtOAc). All pure fractions of 2a were concentrated together to afford yellow oil. Upon standing for at least 24 hours, this oil slowly formed a yellow solid. Analysis of the yellow oil using chiral HPLC revealed that the product was formed in a 68:32 enantiomeric ratio. HPLC condition: Chiralpak AD-H, 80:20, Hexanes:i-PrOH, rt, retention time=33.77 min (major), 37.71 min (minor).
X-Ray Crystallographic Data
The X-ray crystallographic data depicted in
ORTEP structure for compound 1a (
ORTEP Structure for compound 2a (
ORTEP Structure for compound 2o (
ORTEP Structure for compound 2za (
NMR Spectra of the Starting Materials and Products
Please see
The presence of significant signals for residual water in the 1H NMR spectra of 2a, 2e, 2l, 2o, 2t and 2v may raise questions about proof of purity. It should be noted that the presence of one or more heavy halogen atoms and/or an unprotected indole nitrogen atom rendered these compounds only sparingly soluble in common deuterated NMR solvents (CDCl3, CD2Cl2, C6D6, C3D6O, CD3OD, and C2D6SO). Consequently, minor amounts of water contaminant in these solvents may appear to have anomalously enhanced signals relative to those of the compounds of interest. 1H NMR spectra of these compounds were not representative of the bulk material, and cannot be used as the sole judge or proof of purity.
However, the following observations from data presented above were reasonable proof of purity, apart from the 1H-NMR spectra presented in the next section: (i) reasonable accuracy of experimental elemental analysis (C, H, and N) compared to theoretical value (0.05-0.34%) in those cases where combustion analysis was performed; (ii) sharp (1-3° C.) and high (>100° C.) melting points; (iii) substantially higher melting points compared to reported literature value (2a and 2b) [Jessing, M.; Barran, P. S. Heterocycles 2011, 82, 1739-1745]; and, high degree of crystallinity, and with X-ray crystal structures and measurements for 2a, 2o, and 2za.
A model of a Respiratory Syncytial Virus (RSV) L protein was built, using SWISS-MODEL (on the internet at swissmodel(dot)expasy(dot)org). Figures obtained using Chimera and Pymol.
RSV L protein is an RNA-directed RNA polymerase.
Molecular dynamic simulations were carried out to refine the model using NAMD on Graham clusters (Compute Canada). Molecular docking experiments were performed using the Schrödinger Small Molecule Discovery Suite and figures obtained using Pymol. The results are depicted in
(1) (a) Padwa, A.; Weingarten, M. D. Chem. Rev. 1996 96,223-270. (b) Doyle, M. P.; McKervey, M. A.; Ye, T. Modern Catalytic Methods for Organic Synthesis with Diazo Compounds: From Cyclopropanes to Ylides; Wiley: New York, 1998.
(2) (a) Padwa, A.; Hornbuckle, S. F. Chem. Rev. 1991, 91, 263-309. (b) Vanecko, J. A.; Wan, H.; West, F. G.; Tetrahedron 2006, 62, 1043-1062. (c) Murphy, G. K.; Stewart, C.; West, F. G. Tetrahedron 2013, 69, 2667-2686.
(3) Bott, T. M.; Atienza, B. J.; West, F. G. RSC Adv. 2014, 4, 31955-31959.
(4) Liu, J. F.; Jiang, Z. Y.; Wang, R. R.; Zheng, Y. T.; Chen, J. J.; Zhang, X. M.; Ma, Y. B. Org. Lett. 2007, 9, 4127-4129.
(5) (a) Karadeolian, A.; Kerr, M. A. J. Org. Chem. 2010, 75, 6830-6841. (b) Karadeolian, A.; Kerr, M. A. Angew. Chem. Int. Ed. 2010, 49, 1133-1135. (c) Lee, J.; Panek, J. S. Org. Lett. 2011, 13, 502-505. (d) Lee, J.; Panek, J. S. J.Org. Chem. 2015, 80, 2959-2971. (e) Zhang, X.; Mu, T.; Zhan, F.; Ma, L.; Liang, G. Angew. Chem. Int. Ed. 2011, 50, 6164-6166. (f) Wu, W.; Xiao, M.; Wang, J.; Li, Y.; Xie, Z. 9Org. Lett. 2012, 14, 1624-1627. (g) Patel, P.; Ramana, C. V. I Org. Chem. 2012, 77, 10509-10515. (h) Xiao, M.; Wu, W.; Wei, L.; Jin, X.; Yao, X.; Xie, Z. Tetrahedron 2015, 3705-3714.
(6) Nair, H.; Nokes, D. J.; Gessner, B. D.; Dherani, M.; Madhi, S. A.; Singleton, R. J.; O'Brien, K. L.; Roca, A.; Wright, P. F.; Bruce, N.; Chandran, A.; Theodoratou, E.; Sutanto, A.; Sedyaningsih, E. R.; Ngama, M.; Munywoki, P. K.; Kartasasmita, C.; Simoes, E. A.; Rudan, I.; Weber, M. W.; Campbell, H. Lancet 2010, 375, 1545-1555.
(7) Faria, N. R.; Azevedo Rdo, S.; Kraemer, M. U.; Souza, R.; Cunha, M. S.; Hill, S. C.; Theze, J.; Bonsall, M. B.; Bowden, T. A.; Rissanen, I.; Rocco, I. M.; Nogueira, J. S.; Maeda, A. Y.; Vasami, F. G.; Macedo, F. L.; Suzuki, A.; Rodrigues, S. G.; Cruz, A. C.; Nunes, B. T.; Medeiros, D. B.; Rodrigues, D. S.; Nunes Queiroz, A. L.; da Silva, E. V.; Henriques, D. F.; Travassos da Rosa, E. S.; de Oliveira, C. S.; Martins, L. C.; Vasconcelos, H. B.; Casseb, L. M.; Simith Dde, B.; Messina, J. P.; Abade, L.; Lourenco, J.; Carlos Junior Alcantara, L.; de Lima, M. M.; Giovanetti, M.; Hay, S. I.; de Oliveira, R. S.; Lemos Pda, S.; de Oliveira, L. F.; de Lima, C. P.; da Silva, S. P.; de Vasconcelos, J. M.; Franco, L.; Cardoso, J. F.; Vianez-Junior, J. L.; Mir, D.; Bello, G.; Delatorre, E.; Khan, K.; Creatore, M.; Coelho, G. E.; de Oliveira, W. K.; Tesh, R.; Pybus, O. G.; Nunes, M. R.; Vasconcelos, P. F., Science 2016, 352, 345-349.
(8) Mlakar, J.; Korva, M.; Tul, N.; Popovic, M.; Poljsak-Prijatelj, M.; Mraz, J.; Kolenc, M.; Resman Rus, K.; Vesnaver Vipotnik, T.; Fabjan Vodusek, V.; Vizjak, A.; Pizem, J.; Petrovec, M.; Avsic Zupanc, T. N Engl. J. Med. 2016, 374, 951-958.
(9) Salomon, R. G.; Kochi, J. K. J. Am. Chem. Soc. 1973, 95, 3300-3310.
(10) Despite the high nitrogen content of substrate 1a, we have not observed explosive decomposition under standard conditions. Analysis by DSC and TGA indicate thermal instability above 127° C. (See Supporting Information). Nonetheless, use of a blast shield and appropriate protective wear (earplugs, Kevlar lab coat and gloves) is recommended for gram-scale reactions.
(11) Phipps, R. J.; Grimster, N. P.; Gaunt, M. J. J. Am. Chem. Soc. 2008, 130, 8172-8174.
(12) Ribas, X.; Jackson, D. A.; Donnadieu, B.; Mahia, J.; Parella, T.; Xifra, X.; Hedman, B.; Hodgson, K. O.; Llobet, A.; Stack, T. D. P. Angew. Chem. Int. Ed. 2002, 41, 2991-2994.
(13) When the conditions of Table 1, entry 4 were carried out in the presence of various bases (pyridine, triethylamine, various inorganic bases), no conversion was observed. While these results are consistent with the Bronsted acid requirement, they may also be attributable to inhibition of the initial redox activation step by base coordination to Cu(II).
(14) (a) Aburatani, S.; Uenishi, J. Heterocycles 2008, 75, 1407-1416. (b) Quartarone, G.; Charmet, A. P.; Ronchin, L.; Tortato, C.; Vavasori, A. J. Phys. Org. Chem. 2014, 27, 680-689. (c) Guo, T.; Han, S.-L.; Liu, Y.-C.; Liu, H.-M. Tetrahedron Lett. 2016, 57, 1097-1099.
(15) (a) Suárez-Castillo, O. R.; Mélendez-Rodriguez, M.; Morales-Garcia, A. L.; Cano-Escudero, I. C.; Contreras-Martinez, Y. M. A.; Moreles-Rios, M. S.; Joseph-Nathan, P. Heterocycles 2009, 78, 1463-1476. (b) Xu, X.-H.; Liu, G.-K.; Azuma, A.; Tokunaga, E.; Shibata, N. Org. Lett. 2011, 13, 4854-4857.
(16) For related indole additions to an indolenones catalyzed by chrial Brønsted acid, see: (a) Rueping, M.; Raja, S.; Nunez, A. Adv. Synth. Catal. 2011, 353, 563-568. (b) Yin, Q.; You, S.-L. Chem. Sci 2011, 2, 1344-1348.
(17) Sun, Z.; Pan, Y.; Jiang, S.; Lu, L. Viruses 2013, 5, 211-225.
(18) Chapman, J.; Abbott, E.; Alber, D. G.; Baxter, R. C.; Bithell, S. K.; Henderson, E. A.; Carter, M. C.; Chambers, P.; Chubb, A.; Cockerill, G. S.; Collins, P. L.; Dowdell, V. C.; Keegan, S. J.; Kelsey, R. D.; Lockyer, M. J.; Luongo, C.; Najarro, P.; Pickles, R. J.; Simmonds, M.; Taylor, D.; Tyms, S.; Wilson, L. J.; Powell, K. L. Antimicrob. Agents Chemother. 2007, 51, 3346-3353.
(19) Noton, S. L.; Fearns, R. Virology 2015, 479-480C, 545-554.
(20) Selisko, B.; Wang, C.; Harris, E.; Canard, B. Curr. Opin. Virol. 2014, 9, 74-83.
(21) Jessing, M.; Barran, P. S. Heterocycles 2011, 82, 1739-1745.
(22) Hallak, L. K.; Spillmann, D.; Collins, P. L.; Peeples, M. E. J. Virol. 2000, 74, 10508-10513.
(23) Canter, D. M.; Jackson, R. L.; Perrault, J. Virology 1993, 194, 518-529.
(24) Noton, S. L.; Nagendra, K.; Dunn, E. F.; Mawhorter, M. E.; Yu, Q.; Fearns, R. J. Virol. 2015, 89, 7786-7798.
The embodiments described herein are intended to be examples only. Alterations, modifications and variations can be effected to the particular embodiments by those of skill in the art. The scope of the claims should not be limited by the particular embodiments set forth herein, but should be construed in a manner consistent with the specification as a whole.
All publications, patents and patent applications mentioned in this Specification are indicative of the level of skill those skilled in the art to which this invention pertains and are herein incorporated by reference to the same extent as if each individual publication patent, or patent application was specifically and individually indicated to be incorporated by reference.
The invention being thus described, it will be obvious that the same may be varied in many ways. Such variations are not to be regarded as a departure from the spirit and scope of the invention, and all such modification as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.
This application is a national phase filing under 35 U.S.C. § 371 of PCT/CA2019/050380, filed Mar. 27, 2019, which application claims the benefit of U.S. Provisional Patent Application No. 62/649,220, filed Mar. 28, 2018, which applications are incorporated herein by reference in their entirety.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CA2019/050380 | 3/27/2019 | WO |
| Publishing Document | Publishing Date | Country | Kind |
|---|---|---|---|
| WO2019/183729 | 10/3/2019 | WO | A |
| Number | Name | Date | Kind |
|---|---|---|---|
| 20110028564 | Johansen et al. | Feb 2011 | A1 |
| 20140179637 | Leis et al. | Jun 2014 | A1 |
| Number | Date | Country |
|---|---|---|
| WO 2015154169 | Oct 2015 | WO |
| WO 2017055396 | Apr 2017 | WO |
| Entry |
|---|
| Chien et al. Chem. Pharm. Bull. 33(5) 1843-1848 (1985). |
| Registry No. 53904-10-2, File Registry on STN, Nov. 16, 1984. |
| McClay et al. AMB Expr (2015) 5:38, pp. 1-8. |
| Astolfi, et al.; “Radical intermediates in the peroxidation of indoles”; J. Chem. Soc., Perkin Trans.; vol. 2, pp. 1634-1640 (2001). |
| Atienza, et al.; Dual Catalytic Synthesis of Antiviral Compounds Based on Metallocarbene-Azide Cascade Chemistry; J. Org. Chem.; vol. 83, pp. 6829-6842 (2018). |
| Banerji, et al.; “Metal Reagents in Organic Reactions. Part-VI1,2, Oxidation of Indoles with Thallium(III) Acetate”; J. Indian Chem. Soc.; vol. 75, pp. 698-704 (1998). |
| Bayindir, et al.; “A facile one-pot method to synthesise 2-alkylated indole and 2,2′-bis(indolyl)methane derivatives using ketones as electrophiles and their anion sensing ability”; RSC Adv.; vol. 6, pp. 72959-72967 (2016). |
| Berti, et al.; “Electrochemical, Chemical, and E.s.r. Study of the Reduction Mechanism of Substituted Indoxyls in Media with Controlled Proton Availability”; J. Chem. Research; pp. 340-341 (1981). |
| Bott, et al.; “Azide trapping of metallocarbenes: generation of reactive C-acylimines and domino trapping with nucleophiles”; RSC Adv.; vol. 4, pp. 31955-31959 (2014). |
| Colonna, et al.; “Reaction of 2-Phenyl-3H-Indol-3-One With Indoles”; Gazzetta Chimiza Italiana; vol. 105, pp. 985-992 (1975). |
| Ding, et al.; “Photocatalytic aerobic oxidation/semipinacol rearrangement sequence: a concise route to the core of pseudoindoxyl alkaloids”; Tetrahedron Letters; vol. 55, pp. 4648-4652 (2014). |
| Greci, et al.; “Radical cations: reactions of 2-phenylindole with aromatic amines under anodic oxidation. B-Scission of an amino alkoxy radical”; J. Chem. Soc., Perkin Trans. 2; pp. 1749-1755 (2000). |
| Guchhait, et al.; “Oxidative Dearomatization of Indoles via Pd-Catalyzed C—H Oxygenation: An Entry to C2-Quaternary Indolin-3-ones”; Org. Lett.; vol. 18, pp. 1534-1537 (2016). |
| Higuchi, et al.; “Preparation of 2,2-disubstituted 1,2-dihydro-3H-indol-3-ones via oxidation of 2-substituted indoles and Mannich-type reaction”; Tetrahedron; vol. 66, pp. 1236-1243 (2010). |
| Jessing, et al.; “Oxidative Coupling of Indoles with 3-Oxindoles”; Heterocycles; vol. 82, No. 2, pp. 1739-1745 (2011). |
| Li, et al.; “Gold/Copper-Co-catalyzed Tandem Reactions of 2-Alkynylanilines: A Synthetic Strategy for the C2-Quaternary Indolin-3-ones”; Org. Lett.; vol. 19, pp. 1160-1163 (2017). |
| Ling, et al.; Sensitized Photooxidative Coupling of 2-Arylindoles; Youji Huaxue; vol. 6, No. 6, pp. 518-523 (Jan. 1, 1996). |
| Ling, et al.; “Singlet oxygenation of [2,3′-bi-1H-indol]-3(2H)-ones”; Youji Huaxue; vol. 16, No. 2, pp. 152-156 (Jan. 1, 1996). |
| Ling, et al.; “Synthesis of Bulky 2,2-Diaryl-1,2-dihydro-3H-indol-3-ones via Singlet Oxygenation of 2-Arylindoles”; Synthetic Communications; vol. 26, No. 1, pp. 149-152 (1996). |
| Preciado, et al.; “Exploration of Forbidden Povarov Processes as a Source of Unexpected Reactivity: A multicomponent Mannich-Ritter Transformation”; Angew. Chem. Int. Ed.; vol. 51, pp. 6874-6877 (2012). |
| Saito, et al.; “Photoinduced Reactions. LXV. Photosensitized Oxygenation of 2-Methylindoles”; Chemistry Letters; pp. 1173-1176(1972). |
| Yarlagadda, et al.; “Oxidative Asymmetric Aza-Friedel-Crafts Alkylation of Indoles with 3-Indolinone-2-carboxylates Catalyzed by a BINOL Phosphoric Acid and Promoted by DDQ”; Chem. Asian J.; vol. 13, pp. 1327-1334 (2018). |
| Yin, et al.; “Chiral phosphoric acid-catalysed Friedel-Crafts alkylation reaction of indoles with racemic spiro indolin-3-ones”; Chem. Sci.; vol. 2, pp. 1344-1348 (2011). |
| Kong, et al.; “Copper-catalyzed oxidative trimerization of indoles by using TEMPO to construct quaternary carbon centers: the synthesis of 2-(1H-indol-3-yl)-2,3′-biindolin-3-ones”; Canadian Journal of Chemistry; vol. 92, No. 4, pp. 269-273 (Dec. 5, 2013). |
| Lin, et al.; “Silver-catalyzed TEMPO oxidative homocoupling of indoles for the synthesis of 3,3′-biindolin-2-ones”; RSC Advances, vol. 5, No. 46, pp. 37018-37022 (Apr. 13, 2015). |
| Mutule, et al.; “Catalytic Direct Acetoxylation of Indoles”; The Journal of Organic Chemistry; vol. 74, No. 18, pp. 7195-7198 (Aug. 18, 2009). |
| Patterson, et al.; “319. Isatogens. Part II. 2-2′-Pyridylisatogen”, Journal of the Chemical Society; pp. 1706-1711 (Jan. 1, 1965). |
| Shimizu, et al.; “Synthesis of Indolin-3-ones and Tetrahydro-4-quinolones from [alpha]-Imino Esters”; Asian Journal of Organic Chemistry, vol. 2, No. 2, pp. 130-134 (Jan. 17, 2013). |
| Zhang, et al.; “Palladium-Catalyzed One-Pot Synthesis of C2-Quaternary Indolin-3-ones via 1H-indole-3-sulfonates Generated in Situ from 2-Alkynyl Arylazides and Sulfonic Acids”; Advanced Synthesis and Catalysis; vol. 359, No. 23, pp. 4147-4152 (Oct. 4, 2017). |
| Zhou, et al.; “Ruthenium-Catalyzed Oxidative Dearomatization of Indoles for the Construction of C2-Quaternary Indolin-3-ones”; SYNLETT; vol. 29, No. 06, pp. 835-839 (Jan. 15, 2018). |
| Number | Date | Country | |
|---|---|---|---|
| 20210009563 A1 | Jan 2021 | US |
| Number | Date | Country | |
|---|---|---|---|
| 62649220 | Mar 2018 | US |
